Sample records for alkaline phosphate alp

  1. Increased Serum Alkaline Phosphatase and Serum Phosphate as Predictors of Mortality after Stroke

    PubMed Central

    S, Pratibha; JB, Agadi

    2014-01-01

    Context: Serum Alkaline phosphatase (ALP) & phosphate are considered to be indicators of vascular calcification. Link between bone metabolism, vascular calcification, cardiovascular events have been well studied in chronic kidney disease and ischemic heart disease. Aims: To determine that increased serum phosphate and alkaline phosphatase are predictors of mortality rates and recurrent vascular events in stroke. Materials and Methods: Sixty patients admitted with acute stroke (ischemic & haemorrhagic) were included in the study. Their baseline clinical characteristics and biochemical parameters including serum ALP and phosphate were noted. All patients were followed up for a period of one year. The all- cause mortality, the mortality due to cardiovascular events and recurrent vascular events without death were noted during the follow up. Statistical analyses were done to look for any correlation between mortality and baseline levels of serum ALP and phosphate. Results: Of the 60 patients, 8 (13.3%) patients were lost for follow up. Fourteen (26.9%) patients died; of which 12 deaths were due to vascular causes and 2 deaths were due to non vascular causes. Increasing levels of serum ALP and phosphate correlated with all cause mortality and recurrent vascular events without death Conclusion: Serum ALP and phosphate prove to be cost effective prognostic indicator of mortality and recurrent vascular events in stroke. This finding has to be confirmed with studies including larger population. Further research on ALP inhibitors, Vitamin D analogues and phosphate binders to improve mortality in stroke population can be encouraged. PMID:25300293

  2. Increased serum alkaline phosphatase and serum phosphate as predictors of mortality after stroke.

    PubMed

    S, Pratibha; S, Praveen-Kumar; Jb, Agadi

    2014-08-01

    Serum Alkaline phosphatase (ALP) & phosphate are considered to be indicators of vascular calcification. Link between bone metabolism, vascular calcification, cardiovascular events have been well studied in chronic kidney disease and ischemic heart disease. To determine that increased serum phosphate and alkaline phosphatase are predictors of mortality rates and recurrent vascular events in stroke. Sixty patients admitted with acute stroke (ischemic & haemorrhagic) were included in the study. Their baseline clinical characteristics and biochemical parameters including serum ALP and phosphate were noted. All patients were followed up for a period of one year. The all- cause mortality, the mortality due to cardiovascular events and recurrent vascular events without death were noted during the follow up. Statistical analyses were done to look for any correlation between mortality and baseline levels of serum ALP and phosphate. Of the 60 patients, 8 (13.3%) patients were lost for follow up. Fourteen (26.9%) patients died; of which 12 deaths were due to vascular causes and 2 deaths were due to non vascular causes. Increasing levels of serum ALP and phosphate correlated with all cause mortality and recurrent vascular events without death Conclusion: Serum ALP and phosphate prove to be cost effective prognostic indicator of mortality and recurrent vascular events in stroke. This finding has to be confirmed with studies including larger population. Further research on ALP inhibitors, Vitamin D analogues and phosphate binders to improve mortality in stroke population can be encouraged.

  3. Fluorescent Biosensor for Phosphate Determination Based on Immobilized Polyfluorene-Liposomal Nanoparticles Coupled with Alkaline Phosphatase.

    PubMed

    Kahveci, Zehra; Martínez-Tomé, Maria José; Mallavia, Ricardo; Mateo, C Reyes

    2017-01-11

    This work describes the development of a novel fluorescent biosensor based on the inhibition of alkaline phosphatase (ALP). The biosensor is composed of the enzyme ALP and the conjugated cationic polyfluorene HTMA-PFP. The working principle of the biosensor is based on the fluorescence quenching of this polyelectrolyte by p-nitrophenol (PNP), a product of the hydrolysis reaction of p-nitrophenyl phosphate (PNPP) catalyzed by ALP. Because HTMA-PFP forms unstable aggregates in buffer, with low fluorescence efficiency, previous stabilization of the polyelectrolyte was required before the development of the biosensor. HTMA-PFP was stabilized through its interaction with lipid vesicles to obtain stable blue-emitting nanoparticles (NPs). Fluorescent NPs were characterized, and the ability to be quenched by PNP was evaluated. These nanoparticles were coupled to ALP and entrapped in a sol-gel matrix to produce a biosensor that can serve as a screening platform to identify ALP inhibitors. The components of the biosensor were examined before and after sol-gel entrapment, and the biosensor was optimized to allow the determination of phosphate ion in aqueous medium.

  4. Fast and sensitive near-infrared fluorescent probes for ALP detection and 3d printed calcium phosphate scaffold imaging in vivo.

    PubMed

    Park, Chul Soon; Ha, Tai Hwan; Kim, Moonil; Raja, Naren; Yun, Hui-Suk; Sung, Mi Jeong; Kwon, Oh Seok; Yoon, Hyeonseok; Lee, Chang-Soo

    2018-05-15

    Alkaline phosphatase (ALP) is a critical biological marker for osteoblast activity during early osteoblast differentiation, but few biologically compatible methods are available for its detection. Here, we describe the discovery of highly sensitive and rapidly responsive novel near-infrared (NIR) fluorescent probes (NIR-Phos-1, NIR-Phos-2) for the fluorescent detection of ALP. ALP cleaves the phosphate group from the NIR skeleton and substantially alters its photophysical properties, therefore generating a large "turn-on" fluorescent signal resulted from the catalytic hydrolysis on fluorogenic moiety. Our assay quantified ALP activity from 0 to 1.0UmL -1 with a 10 -5 -10 -3 UmL -1 limit of detection (LOD), showing a response rate completed within 1.5min. A potentially powerful approach to probe ALP activity in biological systems demonstrated real-time monitoring using both concentration- and time-dependent variations of endogenous ALP in live cells and animals. Based on high binding affinity to bone tissue of phosphate moiety, bone-like scaffold-based ALP detection in vivo was accessed using NIR probe-labeled three-dimensional (3D) calcium deficient hydroxyapatite (CDHA) scaffolds. They were subcutaneously implanted into mice and monitored ALP signal changes using a confocal imaging system. Our results suggest the possibility of early-stage ALP detection during neo-bone formation inside a bone defect, by in vivo fluorescent evaluation using 3D CDHA scaffolds. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Sensitive detection of alkaline phosphatase by switching on gold nanoclusters fluorescence quenched by pyridoxal phosphate.

    PubMed

    Halawa, Mohamed Ibrahim; Gao, Wenyue; Saqib, Muhammad; Kitte, Shimeles Addisu; Wu, Fengxia; Xu, Guobao

    2017-09-15

    In this work, we designed highly sensitive and selective luminescent detection method for alkaline phosphatase using bovine serum albumin functionalized gold nanoclusters (BSA-AuNCs) as the nanosensor probe and pyridoxal phosphate as the substrate of alkaline phosphatase. We found that pyridoxal phosphate can quench the fluorescence of BSA-AuNCs and pyridoxal has little effect on the fluorescence of BSA-AuNCs. The proposed mechanism of fluorescence quenching by PLP was explored on the basis of data obtained from high-resolution transmission electron microscopy (HRTEM), dynamic light scattering (DLS), UV-vis spectrophotometry, fluorescence spectroscopy, fluorescence decay time measurements and circular dichroism (CD) spectroscopy. Alkaline phosphatase catalyzes the hydrolysis of pyridoxal phosphate to generate pyridoxal, restoring the fluorescence of BSA-AuNCs. Therefore, a recovery type approach has been developed for the sensitive detection of alkaline phosphatase in the range of 1.0-200.0U/L (R 2 =0.995) with a detection limit of 0.05U/L. The proposed sensor exhibit excellent selectivity among various enzymes, such as glucose oxidase, lysozyme, trypsin, papain, and pepsin. The present switch-on fluorescence sensing strategy for alkaline phosphatase was successfully applied in human serum plasma with good recoveries (100.60-104.46%), revealing that this nanosensor probe is a promising tool for ALP detection. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Effect of aluminum phosphate on alkaline phosphatase activity of polyurethane foam immobilized cyanobacteria.

    PubMed

    Ramalingam, N; Prasanna, B Gowtham

    2006-09-01

    The impact of insoluble phosphorus such as aluminum and rock phosphate on alkaline phosphatase activity of polyurethane foam immobilized cyanobacteria was assessed. Polyurethane foam immobilized Nodularia recorded the highest alkaline phosphatase activity of 9.04 (m. mol p-nitrophenol released h(-1) mg(-1) protein) in vitro. A higher concentration of aluminum phosphate was recorded a 25% reduction in alkaline phosphatase activity, ammonia content, and available phosphorus in culture filtrate of polyurethane foam immobilized cyanobacteria. In general, immobilized cyanobacteria exhibited a higher alkaline phosphatase activity in rock phosphate than aluminum phosphate.

  7. Pyrophosphate as substrate for alkaline phosphatase activity: A convenient flow-injection chemiluminescence assay.

    PubMed

    Zhang, Qingfeng; Zhang, Cuiyun; Yang, Meiding; Yu, Donghong; Yu, Cong

    2017-11-01

    A sensitive and convenient flow-injection chemiluminescence (FI-CL) turn-on assay for alkaline phosphatase (ALP) activity without any label and synthesis is developed. Cu 2+ can catalyze the luminol-H 2 O 2 CL reaction. Pyrophosphate (PPi) can chelate Cu 2+ and therefore the Cu 2+ -mediated luminol-H 2 O 2 CL reaction is inhibited. The addition of ALP can catalyze the hydrolysis of PPi into phosphate ions, Cu 2+ is released and the chemiluminescence recovers. A detection limit of 1 mU/mL ALP is obtained. Copyright © 2017 John Wiley & Sons, Ltd.

  8. Chemical redox modulated fluorescence of nitrogen-doped graphene quantum dots for probing the activity of alkaline phosphatase.

    PubMed

    Liu, JingJing; Tang, Duosi; Chen, Zhitao; Yan, Xiaomei; Zhong, Zhou; Kang, Longtian; Yao, Jiannian

    2017-08-15

    Alkaline phosphatase (ALP) as an essential enzyme plays an important role in clinical diagnoses and biomedical researches. Hence, the development of convenient and sensitivity assay for monitoring ALP is extremely important. In this work, on the basis of chemical redox strategy to modulate the fluorescence of nitrogen-doped graphene quantum dots (NGQDs), a novel label-free fluorescent sensing system for the detection of alkaline phosphatase (ALP) activity has been developed. The fluorescence of NGQDs is firstly quenched by ultrathin cobalt oxyhydroxide (CoOOH) nanosheets, and then restored by ascorbic acid (AA), which can reduce CoOOH to Co 2+ , thus the ALP can be monitored based on the enzymatic hydrolysis of L-ascorbic acid-2-phosphate (AAP) by ALP to generate AA. Quantitative evaluation of ALP activity in a range from 0.1 to 5U/L with the detection limit of 0.07U/L can be realized in this sensing system. Endowed with high sensitivity and selectivity, the proposed assay is capable of detecting ALP in biological system with satisfactory results. Meanwhile, this sensing system can be easily extended to the detection of various AA-involved analytes. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. ESTIMATION OF PHOSPHATE ESTER HYDROLYSIS RATE CONSTANTS - ALKALINE HYDROLYSIS

    EPA Science Inventory

    SPARC (SPARC Performs Automated Reasoning in Chemistry) chemical reactivity models were extended to allow the calculation of alkaline hydrolysis rate constants of phosphate esters in water. The rate is calculated from the energy difference between the initial and transition state...

  10. Imaging of Alkaline Phosphatase Activity in Bone Tissue

    PubMed Central

    Gade, Terence P.; Motley, Matthew W.; Beattie, Bradley J.; Bhakta, Roshni; Boskey, Adele L.; Koutcher, Jason A.; Mayer-Kuckuk, Philipp

    2011-01-01

    The purpose of this study was to develop a paradigm for quantitative molecular imaging of bone cell activity. We hypothesized the feasibility of non-invasive imaging of the osteoblast enzyme alkaline phosphatase (ALP) using a small imaging molecule in combination with 19Flourine magnetic resonance spectroscopic imaging (19FMRSI). 6, 8-difluoro-4-methylumbelliferyl phosphate (DiFMUP), a fluorinated ALP substrate that is activatable to a fluorescent hydrolysis product was utilized as a prototype small imaging molecule. The molecular structure of DiFMUP includes two Fluorine atoms adjacent to a phosphate group allowing it and its hydrolysis product to be distinguished using 19Fluorine magnetic resonance spectroscopy (19FMRS) and 19FMRSI. ALP-mediated hydrolysis of DiFMUP was tested on osteoblastic cells and bone tissue, using serial measurements of fluorescence activity. Extracellular activation of DiFMUP on ALP-positive mouse bone precursor cells was observed. Concurringly, DiFMUP was also activated on bone derived from rat tibia. Marked inhibition of the cell and tissue activation of DiFMUP was detected after the addition of the ALP inhibitor levamisole. 19FMRS and 19FMRSI were applied for the non-invasive measurement of DiFMUP hydrolysis. 19FMRS revealed a two-peak spectrum representing DiFMUP with an associated chemical shift for the hydrolysis product. Activation of DiFMUP by ALP yielded a characteristic pharmacokinetic profile, which was quantifiable using non-localized 19FMRS and enabled the development of a pharmacokinetic model of ALP activity. Application of 19FMRSI facilitated anatomically accurate, non-invasive imaging of ALP concentration and activity in rat bone. Thus, 19FMRSI represents a promising approach for the quantitative imaging of bone cell activity during bone formation with potential for both preclinical and clinical applications. PMID:21799916

  11. ESTIMATION OF PHOSPHATE ESTER HYDROLYSIS RATE CONSTANTS. I. ALKALINE HYDROLYSIS

    EPA Science Inventory

    SPARC (SPARC Performs Automated Reasoning in Chemistry) chemical reactivity models were extended to allow the calculation of alkaline hydrolysis rate constants of phosphate esters in water. The rate is calculated from the energy difference between the initial and transition state...

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

    PubMed

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

    2006-07-12

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

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

    PubMed

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

    2015-06-09

    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. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Construction of an alkaline phosphatase-specific two-photon probe and its imaging application in living cells and tissues.

    PubMed

    Zhang, Huatang; Xiao, Peng; Wong, Yin Ting; Shen, Wei; Chhabra, Mohit; Peltier, Raoul; Jiang, Yin; He, Yonghe; He, Jun; Tan, Yi; Xie, Yusheng; Ho, Derek; Lam, Yun-Wah; Sun, Jinpeng; Sun, Hongyan

    2017-09-01

    Alkaline phosphatase (ALP) is a family of enzymes involved in the regulation of important biological processes such as cell differentiation and bone mineralization. Monitoring the activity of ALP in serum can help diagnose a variety of diseases including bone and liver diseases. There has been growing interest in developing new chemical tools for monitoring ALP activity in living systems. Such tools will help further delineate the roles of ALP in biological and pathological processes. Previously reported fluorescent probes has a number of disadvantages that limit their application, such as poor selectivity and short-wavelength excitation. In this work, we report a new two-photon fluorescent probe (TP-Phos) to selectively detect ALP activity. The probe is composed of a two-photon fluorophore, a phosphate recognition moiety, and a self-cleavable adaptor. It offers a number of advantages over previously reported probes, such as fast reaction kinetics, high sensitivity and low cytotoxicity. Experimental results also showed that TP-Phos displayed improved selectivity over DIFMUP, a commonly utilized ALP probe. The selectivity is attributed to the utilization of an ortho-functionalised phenyl phosphate group, which increases the steric hindrance of the probe and the active site of phosphatases. Moreover, the two-photon nature of the probe confers enhanced imaging properties such as increased penetration depth and lower tissue autofluorescence. TP-Phos was successfully used to image the endogenous ALP activity of hippocampus, kidney and liver tissues from rat. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Differential Properties of Human ALP+ Periodontal Ligament Stem Cells vs Their ALP- Counterparts

    PubMed Central

    Tran, Quynh T; El-Ayachi, Ikbale; Bhatti, Fazal-Ur-Rehman; Bahabri, Rayan; Al-Habib, Mey; Huang, George TJ

    2015-01-01

    Characterizing subpopulations of stem cells is important to understand stem cell properties. Tissue-nonspecific alkaline phosphatase (ALP) is associated with mineral tissue forming cells as well as stem cells. Information regarding ALP subpopulation of human periodontal ligament stem cells (hPDLSCs) is limited. In the present study, we examined ALP+ and ALP− hPDLSC subpopulations, their surface markers STRO-1 and CD146, and the expression of stemness genes at various cell passages. We found that ALP+ subpopulation had higher levels of STRO-1 (30.6 ± 5.6%) and CD146 (90.4 ± 3.3%) compared to ALP− (STRO-1: 0.5 ± 0.1%; CD146: 75.3 ± 7.2%). ALP+ cells expressed significantly higher levels of stemness associated genes, NANOG, OCT4 and SOX than ALP− cells at low cell passages of 2-3 (p<0.05). ALP+ and ALP− cells had similar osteogenic, chondrogenic and neurogenic potential while ALP−, not ALP+ cells, lacked adipogenic potential. Upon continuous culturing and passaging, ALP+ continued to express higher stemness genes and STRO-1 and CD146 than ALP− cells at ≥passage 19. Under conditions (over-confluence and vitamin C treatment) when ALP+ subpopulation was increased, the stemness gene levels of ALP+ was no longer significantly higher than those in ALP− cells. In conclusion, ALP+ hPDLSCs possess differential properties from their ALP− counterparts. PMID:26807329

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

    PubMed

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

    2016-01-01

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

  17. Carbon quantum dots-based recyclable real-time fluorescence assay for alkaline phosphatase with adenosine triphosphate as substrate.

    PubMed

    Qian, Zhaosheng; Chai, Lujing; Tang, Cong; Huang, Yuanyuan; Chen, Jianrong; Feng, Hui

    2015-03-03

    A convenient, reliable, and highly sensitive real-time assay for alkaline phosphatase (ALP) activity in the continuous and recyclable way is established on the basis of aggregation and disaggregation of carbon quantum dots (CQDs) through the competitive assay approach. CQDs and adenosine triphosphate (ATP) were used as the fluorescent indicator and substrate for ALP activity assessment, respectively. Richness of carboxyl groups on the surface of CQDs enables their severe aggregation triggered by cerium ions, which results in effective fluorescence quenching. Under the catalytic hydrolysis of ALP, ATP can be rapidly transformed to phosphate ions. Stronger affinity of phosphate ions to cerium ions than carboxyl groups is taken advantage of to achieve fluorescence recovery induced by redispersion of CQDs in the presence of ALP and ATP. Quantitative evaluation of ALP activity in a broad range from 4.6 to 383.3 U/L with the detection limit of 1.4 U/L can be realized in this way, which endows the assay with high enough sensitivity for practical detection in human serum. The assay can be used in a recyclable way for more than three times since the generated product CePO4 as a precipitate can be easily removed from the standard assay system. This strategy broadens the sensing application of fluorescent CQDs with excellent biocompatibility and provides an example based on disaggregation in optical probe development.

  18. Alkaline resistant phosphate glasses and method of preparation and use thereof

    DOEpatents

    Brow, Richard K.; Reis, Signo T.; Velez, Mariano; Day, Delbert E.

    2010-01-26

    A substantially alkaline resistant calcium-iron-phosphate (CFP) glass and methods of making and using thereof. In one application, the CFP glass is drawn into a fiber and dispersed in cement to produce glass fiber reinforced concrete (GFRC) articles having the high compressive strength of concrete with the high impact, flexural and tensile strength associated with glass fibers.

  19. Enhancement of lateral flow immunoassay by alkaline phosphatase: a simple and highly sensitive test for potato virus X.

    PubMed

    Panferov, Vasily G; Safenkova, Irina V; Varitsev, Yury A; Zherdev, Anatoly V; Dzantiev, Boris B

    2017-12-06

    Alkaline phosphatase (ALP) was used as an amplification tool in lateral flow immunoassay (LFIA). Potato virus Х (PVX) was selected as a target analyte because of its high economic importance. Two conjugates of gold nanoparticles were applied, one with mouse monoclonal antibody against PVX and one with ALP-labeled antibody against mouse IgG. They were immobilized to two fiberglass membranes on the test strip for use in LFIA. After exposure to the sample, a substrate for ALP (5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium) was dropped on the test strip. The insoluble dark-violet diformazan produced by ALP precipitated on the membrane and significantly increased the color intensity of the control and test zones. The limit of detection (0.3 ng mL -1 ) was 27 times lower than that of conventional LFIA for both buffer and potato leaf extracts. The ALP-enhanced LFIA does not require additional preparation procedures or washing steps and may be used by nontrained persons in resource-limited conditions. The new method of enhancement is highly promising and may lead to application for routine LFIA in different areas. Graphical abstract Two gold nanoparticles (GNP) conjugates were used - the first with monoclonal antibodies (mAb) (GNP-mAb); the second - alkaline phosphatase-labeled antibody against mAb (GNP-anti-mAb-ALP). The immuno complexes are captured by the polyclonal antibodies (pAb) in the test zone. Addition of the substrate solution (BCIP/NBT) results in the accumulation of the insoluble colored product and in a significance increase in color intensity.

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

    PubMed

    Fenton, Tanis R; Lyon, Andrew W; Rose, M Sarah

    2011-08-31

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

  1. Sequence and functional analysis of intestinal alkaline phosphatase from Lateolabrax maculatus.

    PubMed

    Wu, Minglin; Wang, Jiaqi; Wang, Zhipeng; Zhao, Jinliang; Hu, Yuting; Chen, Xiaowu

    2017-12-01

    Alkaline phosphatases (Alps) belong to a class of phosphate transferases that dephosphorylate lipopolysaccharide (LPS), adenosine triphosphate, and nucleotides. In this study, a 1874-base pair (bp) intestinal alp cDNA sequence was cloned from Lateolabrax maculatus and designated as Lm-alpi. It contained a 1611 bp open reading frame which encoded a protein with 537 amino acids. Protein sequence alignment showed that Lm-AlpI shared 29.8-79.8% identity with its homologs. Lm-AlpI catalytic sites contained three metal ion sites (two Zn 2+ and one Mg 2+ ), referring to D73, H184, D348, H349, H352, H464, D389, and H390 residues, which are essential for enzymatic activity and conservation in different organisms. Two predicted disulfide bonds in Lm-AlpI were composed of four cysteines (C152-C214 and C499-C506), which were homologous to those of mammals. Immunohistochemical staining revealed that Lm-AlpI was mainly expressed on the mucosal surface of the gastrointestinal tract, including stomach, intestine, and gastric cecum. Lm-AlpI was mainly located on the plasma membrane of transiently transfected HeLa cells. The mRNA of Lm-alpi was mainly expressed in the intestine, and its expression levels gradually increased after LPS treatment and further increased by 1.81-fold after 48 h. After desalting culture, the relative mRNA expression level of Lm-alpi decreased at 30 and 50 days after hatching (DAH) and then returned to normal levels at 70 DAH. Further experiments demonstrated that the enzyme activity of Lm-AlpI exhibited an expression pattern similar to that of the mRNA expression of Lm-alpi after LPS treatment and desalting culture. This study provided valuable information on the Lm-AlpI functions associated with the mucosal immunity and salinity adaptation of L. maculatus.

  2. Glycation Contributes to Interaction Between Human Bone Alkaline Phosphatase and Collagen Type I.

    PubMed

    Halling Linder, Cecilia; Enander, Karin; Magnusson, Per

    2016-03-01

    Bone is a biological composite material comprised primarily of collagen type I and mineral crystals of calcium and phosphate in the form of hydroxyapatite (HA), which together provide its mechanical properties. Bone alkaline phosphatase (ALP), produced by osteoblasts, plays a pivotal role in the mineralization process. Affinity contacts between collagen, mainly type II, and the crown domain of various ALP isozymes were reported in a few in vitro studies in the 1980s and 1990s, but have not attracted much attention since, although such interactions may have important implications for the bone mineralization process. The objective of this study was to investigate the binding properties of human collagen type I to human bone ALP, including the two bone ALP isoforms B1 and B2. ALP from human liver, human placenta and E. coli were also studied. A surface plasmon resonance-based analysis, supported by electrophoresis and blotting, showed that bone ALP binds stronger to collagen type I in comparison with ALPs expressed in non-mineralizing tissues. Further, the B2 isoform binds significantly stronger to collagen type I in comparison with the B1 isoform. Human bone and liver ALP (with identical amino acid composition) displayed pronounced differences in binding, revealing that post-translational glycosylation properties govern these interactions to a large extent. In conclusion, this study presents the first evidence that glycosylation differences in human ALPs are of crucial importance for protein-protein interactions with collagen type I, although the presence of the ALP crown domain may also be necessary. Different binding affinities among the bone ALP isoforms may influence the mineral-collagen interface, mineralization kinetics, and degree of bone matrix mineralization, which are important factors determining the material properties of bone.

  3. Pho4 Is Essential for Dissemination of Cryptococcus neoformans to the Host Brain by Promoting Phosphate Uptake and Growth at Alkaline pH

    PubMed Central

    Kaufman-Francis, Keren; Desmarini, Desmarini; Juillard, Pierre G.; Li, Cecilia; Stifter, Sebastian A.; Feng, Carl G.; Sorrell, Tania C.; Grau, Georges E. R.; Bahn, Yong-Sun

    2017-01-01

    ABSTRACT Phosphate acquisition by fungi is regulated by the phosphate-sensing and acquisition (PHO) signaling pathway. Cryptococcus neoformans disseminates from the lung to the brain and is the commonest cause of fungal meningitis worldwide. To investigate the contribution of PHO signaling to cryptococcal dissemination, we characterized a transcription factor knockout strain (hlh3Δ/pho4Δ) defective in phosphate acquisition. Despite little similarity with other fungal Pho4 proteins, Hlh3/Pho4 functioned like a typical phosphate-responsive transcription factor in phosphate-deprived cryptococci, accumulating in nuclei and triggering expression of genes involved in phosphate acquisition. The pho4Δ mutant strain was susceptible to a number of stresses, the effect of which, except for alkaline pH, was alleviated by phosphate supplementation. Even in the presence of phosphate, the PHO pathway was activated in wild-type cryptococci at or above physiological pH, and under these conditions, the pho4Δ mutant had a growth defect and compromised phosphate uptake. The pho4Δ mutant was hypovirulent in a mouse inhalation model, where dissemination to the brain was reduced dramatically, and markedly hypovirulent in an intravenous dissemination model. The pho4Δ mutant was not detected in blood, nor did it proliferate significantly when cultured with peripheral blood monocytes. In conclusion, dissemination of infection and the pathogenesis of meningitis are dependent on cryptococcal phosphate uptake and stress tolerance at alkaline pH, both of which are Pho4 dependent. IMPORTANCE Cryptococcal meningitis is fatal without treatment and responsible for more than 500,000 deaths annually. To be a successful pathogen, C. neoformans must obtain an adequate supply of essential nutrients, including phosphate, from various host niches. Phosphate acquisition in fungi is regulated by the PHO signaling cascade, which is activated when intracellular phosphate decreases below a critical

  4. [Osteogenic activity of porous calcium phosphate ceramics fabricated by rapid prototyping].

    PubMed

    He, Chenguang; Zhao, Li; Lin, Liulan; Gu, Huijie; Zhou, Heng; Cui, Lei

    2010-07-01

    Calcium phosphate bioceramics has a broad application prospect because of good biocompatibility, but porous scaffolds with complex shape can not be prepared by the traditional methods. To fabricate porous calcium phosphate ceramics by rapid prototyping and to investigate the in vitro osteogenic activities. The porous calcium phosphate ceramics was fabricated by rapid prototyping. The bone marrow mesenchymal stem cells (BMSCs) were isolated from bone marrow of Beagle canine, and the 3rd passage BMSCs were seeded onto the porous ceramics. The cell/ceramics composite cultured in osteogenic medium were taken as the experimental group (group A) and the cell/ceramics composite cultured in growth medium were taken as the control group (group B). Meanwhile, the cells seeded on the culture plate were cultured in osteogenic medium or growth medium respectively as positive control (group C) or negative control (group D). After 1, 3, and 7 days of culture, the cell proliferation and osteogenic differentiation on the porous ceramics were evaluated by DNA quantitative analysis, histochemical staining and alkaline phosphatase (ALP) activity. After DiO fluorescent dye, the cell adhesion, growth, and proliferation on the porous ceramics were also observed by confocal laser scanning microscope (CLSM). DNA quantitative analysis results showed that the number of BMSCs in all groups increased continuously with time. Plateau phase was not obvious in groups A and B, but it was clearly observed in groups C and D. The CLSM observation indicated that the activity of BMSCs was good and the cells spread extensively, showing good adhesion and proliferation on the porous calcium phosphate ceramics prepared by rapid prototyping. ALP quantitative analysis results showed that the stain of cells on the ceramics became deeper and deeper with time in groups A and B, the staining degree in group A were stronger than that in group B. There was no significant difference in the change of the ALP activity

  5. Iodine-Mediated Etching of Gold Nanorods for Plasmonic ELISA Based on Colorimetric Detection of Alkaline Phosphatase.

    PubMed

    Zhang, Zhiyang; Chen, Zhaopeng; Wang, Shasha; Cheng, Fangbin; Chen, Lingxin

    2015-12-23

    Here, we propose a plasmonic enzyme-linked immunosorbent assay (ELISA) based on highly sensitive colorimetric detection of alkaline phosphatase (ALP), which is achieved by iodine-mediated etching of gold nanorods (AuNRs). Once the sandwich-type immunocomplex is formed, the ALP bound on the polystyrene microwells will hydrolyze ascorbic acid 2-phosphate into ascorbic acid. Subsequently, iodate is reduced to iodine, a moderate oxidant, which etches AuNRs from rod to sphere in shape. The shape change of AuNRs leads to a blue-shift of longitudinal localized surface plasmon resonance. As a result, the solution of AuNRs changes from blue to red. Benefiting from the highly sensitive detection of ALP, the proposed plasmonic ELISA has achieved an ultralow detection limit (100 pg/mL) for human immunoglobulin G (IgG). Importantly, the visual detection limit (3.0 ng/mL) allows the rapid differential diagnosis with the naked eye. The further detection of human IgG in fetal bovine serum indicates its applicability to the determination of low abundance protein in complex biological samples.

  6. Osteogenic Differentiation of Human and Ovine Bone Marrow Stromal Cells in response to β-Glycerophosphate and Monosodium Phosphate.

    PubMed

    Bottagisio, Marta; Lovati, Arianna B; Lopa, Silvia; Moretti, Matteo

    2015-08-01

    Bone defects are severe burdens in clinics, and thus cell therapy offers an alternative strategy exploiting the features of bone marrow stromal cells (BMSCs). Sheep are a suitable orthopedic preclinical model for similarities with humans. This study compares the influence of two phosphate sources combined with bone morphogenetic protein-2 (BMP-2) on the osteogenic potential of human and ovine BMSCs. β-Glycerophosphate (β-GlyP) and monosodium phosphate (NaH2PO4) were used as organic and inorganic phosphate sources. Osteogenic differentiation of the BMSCs was assessed by calcified matrix, alkaline phosphatase (ALP) activity, and gene expression analysis. A higher calcified matrix deposition was detected in BMSCs cultured with NaH2PO4. Although no significant differences were detected among media for human BMSCs, β-GlyP with or without BMP-2 determined a positive trend in ALP levels compared to NaH2PO4. In contrast, NaH2PO4 had a positive effect on ALP levels in ovine BMSCs. β-GlyP better supported the expression of COL1A1 in human BMSCs, whereas all media enhanced RUNX2 and SPARC expression. Ovine BMSCs responded poorly to any media for RUNX2, COL1A1, and SPARC expression. NaH2PO4 improved calcified matrix deposition without upregulating the transcriptional expression of osteogenic markers. A further optimization of differentiation protocols needs to be performed to translate the procedures from preclinical to clinical models.

  7. Human alkaline phosphatase dephosphorylates microbial products and is elevated in preterm neonates with a history of late-onset sepsis.

    PubMed

    Pettengill, Matthew; Matute, Juan D; Tresenriter, Megan; Hibbert, Julie; Burgner, David; Richmond, Peter; Millán, José Luis; Ozonoff, Al; Strunk, Tobias; Currie, Andrew; Levy, Ofer

    2017-01-01

    A host defense function for Alkaline phosphatases (ALPs) is suggested by the contribution of intestinal ALP to detoxifying bacterial lipopolysaccharide (endotoxin) in animal models in vivo and the elevation of ALP activity following treatment of human cells with inflammatory stimuli in vitro. However the activity of ALP in human plasma (primarily tissue-nonspecific ALP; TNAP) on lipopolysaccharide and other microbial products has not been assessed, nor has its expression been studied in preterm newborns, a vulnerable population at high risk of sepsis. In this context, the aim of our study was to characterize the activity of TNAP on Toll-like receptor (TLR) agonists and assess the concentrations of plasma ALP during late-onset sepsis in preterm newborns. Recombinant human TNAP was incubated with microbial products and phosphate release was measured by malachite green assay. Plasma ALP activity was measured serially in a cohort of preterm (N = 129) infants at high risk of late-onset sepsis (LOS). TNAP dephosphorylates poly-inosine:cytosine (Toll-like receptor (TLR) 3 agonist) and LPS from Klebsiella pneumoniae and Salmonella minnesota (TLR4 agonists). Plasma ALP significantly increased postnatally over the first 4 weeks of life in preterm and term newborns. Bacteremic LOS in preterm infants (gestational age ≤ 30 weeks) was associated with significantly elevated plasma ALP at 4 weeks postnatal age. TNAP, the main circulating isozyme of ALP, de-phosphorylates TLR agonists, demonstrates a post-natal age dependent increase in preterm and term plasma across the first 4 weeks of life, and is elevated in association with preterm LOS.

  8. Association between serum alkaline phosphatase and coronary artery calcification in a sample of primary cardiovascular prevention patients.

    PubMed

    Panh, Loïc; Ruidavets, Jean Bernard; Rousseau, Hervé; Petermann, Antoine; Bongard, Vanina; Bérard, Emilie; Taraszkiewicz, Dorota; Lairez, Olivier; Galinier, Michel; Carrié, Didier; Ferrières, Jean

    2017-05-01

    A high level of serum alkaline phosphatase (ALP) is associated with an increased risk of mortality and myocardial infarction. ALP hydrolyses inorganic pyrophosphate, which is a strong inhibitor of calcium phosphate deposition. The aim of this study was to determine whether ALP is associated with the coronary artery calcium score (CACS). We examined the association of CACS, assessed by computed tomography scanning, and ALP, in 500 patients consecutively recruited, free of cardiovascular disease. The CACS were categorized into two groups: no calcification (CACS = 0) (n = 187) and with calcification (CACS>0) (n = 313). ALP activity was divided into three tertile groups: low ALP level (<55 IU/L), intermediate (55-66 IU/L) and high ALP level (>66 IU/L). The mean age was 60.9 ± 10.8 years, 49.6% of the patients were women. ALP ranged from 22 to 164 IU/L (mean 62.6 IU/L, SD 19.3). In univariate analysis, traditional cardiovascular risk factors, statin use (p = 0.001), and ALP (p = 0.001) were significantly associated with CACS. After adjusting for cardiovascular risk factors, only age (p = 0.001) and sex (p = 0.001) were independently associated with CACS. Compared to the tertile group with low levels of ALP, the intermediate tertile group [OR 2.11, 95% CI (1.12; 3.96), p = 0.02], as well as the high tertile group [OR 3.89, 95% CI (2.01; 7.54), p = 0.001)], was independently associated with CACS. In patients free of cardiovascular disease, high ALP levels are positively and independently associated with coronary artery calcification. The metabolic pathway of ALP and inorganic pyrophosphate could be a target for new therapies against vascular calcification. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Elastase, alpha2-macroglobulin and alkaline phosphatase in crevicular fluid from implants with and without periimplantitis.

    PubMed

    Plagnat, Dominique; Giannopoulou, Catherine; Carrel, Anne; Bernard, Jean-Pierre; Mombelli, Andrea; Belser, Urs C

    2002-06-01

    The aim of this investigation was to determine the presence of selected enzymes and enzyme inhibitors in crevicular fluid collected from implants with and without clinical, radiographic and microbiological signs of periimplantitis. Eleven implants with symptoms of periimplantitis in eight patients (four men and four women) were compared to eleven implants in seven subjects (one man and six women) without periimplantitis. Periimplant crevicular fluid (PICF) was collected at the mesial and distal sites of each implant. Alkaline phosphatase activity (ALP) was measured by using p-nitrophenyl-phosphate as substrate, elastase activity (EA) by the use of a low molecular weight fluorogenic substrate, and the inhibitor alpha2-macroglobulin (alpha2M) by ELISA. ALP, EA and alpha2M were detected in the majority of samples in both groups. In comparison to the clinically healthy implants, total amounts of each of these substances were significantly higher in PICF collected around implants with periimplantitis. The mean total amounts of EA, alpha2M and ALP in the healthy group were: EA: 1.8 ng, alpha2M: 3.1 ng, ALP: 24.1 U, and in the periimplantitis group EA: 23.1 ng, alpha2M: 25.2 ng and ALP: 142.3 U. In addition, all three mediators were correlated with the clinical parameters. The results confirm the similarity of the inflammatory response of tissues surrounding implants and natural teeth, and suggest that ALP and EA could be promising markers of bone loss around dental implants.

  10. Fluorometric determination of the activity of alkaline phosphatase based on the competitive binding of gold nanoparticles and pyrophosphate to CePO4:Tb nanorods.

    PubMed

    Xu, Ai-Zhen; Zhang, Li; Zeng, Hui-Hui; Liang, Ru-Ping; Qiu, Jian-Ding

    2018-05-09

    A fluorometric method is described for the determination of the activity of alkaline phosphatase (ALP). It relies on the competition between gold nanoparticles (AuNPs) and pyrophosphate (PPi) for the coordination sites on the surface of CePO 4 :Tb nanorods. The green fluorescence of the CePO 4 :Tb is reduced in the presence of AuNPs due to fluorescence resonance energy transfer (FRET), but can be restored on addition of PPi due to the stronger affinity of PPi to the CePO 4 :Tb. In the presence of ALP, PPi is hydrolyzed to form phosphate which has much weaker affinity for the CePO 4 :Tb. Hence, the AuNPs will reassemble on the CePO 4 :Tb, and fluorescence is reduced. Fluorescence drops linearly in the 0.2 to 100 U·L -1 activity range, and the detection limit is 60 mU·L -1 (at S/N = 3). The method does not require any modification of the surface of the CePO 4 :Tb and is highly sensitive and selective. The inhibition of ALP activity by Na 3 VO 4 was also studied. In our perception, the method may find application in the diagnosis of ALP-related diseases, in screening for inhibitors, and in studies on ALP-related functions in biological systems. Graphical abstract A assay for the detection of alkaline phosphatase is proposed based on the fluorescence resonance energy transfer between CePO 4 :Tb and AuNPs. It relies on the competitive binding of AuNPs and pyrophosphate (PPi) to CePO 4 :Tb and the hydrolysis of PPi by ALP.

  11. Osteoblast response to zirconia-hybridized pyrophosphate-stabilized amorphous calcium phosphate

    PubMed Central

    Whited, Bryce M.; Skrtic, Drago; Love, Brian J.

    2006-01-01

    Calcium phosphate bioceramics, such as hydroxyapatite, have long been used as bone substitutes because of their proven biocompatibility and bone binding properties in vivo. Recently, a zirconia-hybridized pyrophosphate-stabilized amorphous calcium phosphate (Zr-ACP) has been synthesized, which is more soluble than hydroxyapatite and allows for controlled release of calcium and phosphate ions. These ions have been postulated to increase osteoblast differentiation and mineralization in vitro. The focus of this work is to elucidate the physicochemical properties of Zr-ACP and to measure cell response to Zr-ACP in vitro using a MC3T3-E1 mouse calvarial-derived osteoprogenitor cell line. Cells were cultured in osteogenic medium and mineral was added to culture at different stages in cell maturation. Culture in the presence of Zr-ACP showed significant increases in cell proliferation, alkaline phosphatase activity (ALP), and osteopontin (OPN) synthesis, whereas collagen synthesis was unaffected. In addition, calcium and phosphate ion concentrations and medium pH were found to transiently increase with the addition of Zr-ACP, and are hypothesized to be responsible for the osteogenic effect of Zr-ACP. PMID:16278876

  12. Detection of alkaline phosphatase in canine cells previously stained with Wright-Giemsa and its utility in differentiating osteosarcoma from other mesenchymal tumors.

    PubMed

    Ryseff, Julia K; Bohn, Andrea A

    2012-09-01

    Osteosarcoma (OSA) is a common primary bone tumor in dogs. Demonstration of alkaline phosphatase (ALP) reactivity by tumor cells on unstained slides is useful in differentiating osteosarcoma from other types of sarcoma. However, unstained slides are not always available. The objectives of this study were to evaluate the diagnostic utility of detecting ALP expression in differentiating osteosarcoma from other sarcomas in dogs using cytologic material previously stained with Wright-Giemsa stain and to assess the sensitivity and specificity of ALP expression for diagnosing osteosarcoma using a specific protocol. Archived aspirates of histologically confirmed sarcomas in dogs that had been previously stained with Wright-Giemsa stain were treated with 5-bromo, 4-chloro, 3-indolyl phosphate/nitroblue tetrazolium (BCIP/NBT) as a substrate for ALP. Cells were evaluated for expression of ALP after incubation with BCIP/NBT for 1 hour. Sensitivity and specificity of ALP expression for diagnosis of OSA were calculated. In samples from 83 dogs, cells from 15/17 OSAs and from 4/66 tumors other than OSA (amelanotic melanoma, gastrointestinal stromal tumor, collision tumor, and anaplastic sarcoma) expressed ALP. Sensitivity and specificity of ALP expression detected using BCIP/NBT substrate applied to cells previously stained with Wright-Giemsa stain for OSA were 88 and 94%, respectively. ALP expression detected using BCIP/NBT substrate applied to previously stained cells is useful in differentiating canine OSA from other mesenchymal neoplasms. © 2012 American Society for Veterinary Clinical Pathology.

  13. Temperature enhances the affinity of soil alkaline phosphatase to Cd.

    PubMed

    Tan, Xiangping; Machmuller, Megan B; Wang, Ziquan; Li, Xudong; He, Wenxiang; Cotrufo, M Francesca; Shen, Weijun

    2018-04-01

    Both elevated temperature and heavy metal contamination can have profound effects on microbial function and soil biogeochemical cycling. However, the interactive effects of heavy metal toxicity and temperature on microbial activity have been poorly understood. The aim of this study was to quantify the effect of temperature and cadmium (Cd) toxicity on alkaline phosphatase (ALP) produced by microbes to acquire phosphorus. To determine whether these effects were dependent on soil properties, we utilized 11 soil types from cropland throughout China. We measured ALP activities and kinetics across a temperature (17, 27, 37, and 47 °C) and Cd concentration gradient (0, 0.6, 5, 25, 50, 100, 200, 300, and 500 mg kg -1 ). We found that the half saturation constant (K m ) and the velocity constant (k) of ALP increased nonlinearly with temperature across all soil types. However, the maximum reaction velocity (V max ) increased linearly with temperature. Regardless of soil type and temperature, Cd had a non-competitive inhibitory mechanism. Soil pH, TOC, and clay content were the major factors controlling the affinity of ALP for Cd (K i ). The ecology dose (ED 50 ) for V max and k, and K i were negatively related to temperature, indicating that the toxicity of Cd on ALP is temperature-dependent. Additionally, higher temperatures led to more inhibition of Cd on ALP activity in alkaline soils than that in acidic and neutral soils. Our results suggest that global warming might accelerate the deficiency of available phosphorus in Cd contaminated soils due to higher inhibition of Cd on ALP activity, particularly in alkaline soils. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Probable levetiracetam-related serum alkaline phosphatase elevation

    PubMed Central

    2012-01-01

    Background Levetiracetam (LEV) is an antiepileptic drug with a favorable tolerability and safety profile with little or no effect on liver function. Case presentation Here, we reported an epileptic pediatric patient who developed a significant elevation in serum alkaline phosphatase level (ALP) during LEV monotherapy. Moreover, the serum ALP level was surprisingly decreased to normal after LEV discontinuation. The Naranjo Adverse Drug Reaction Probability Scale score was 6, indicating firstly LEV was a probable cause for the increased serum ALP. Conclusions Cautious usage and concerns of the LEV-associated potential ALP elevation should be considered when levetiracetam is prescribed to epilepsy patients, especially pediatric patients. PMID:22994584

  15. Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli

    NASA Astrophysics Data System (ADS)

    Bozkurt, Akif Goktug; Buyukgoz, Guluzar Gorkem; Soforoglu, Mehmet; Tamer, Ugur; Suludere, Zekiye; Boyaci, Ismail Hakki

    2018-04-01

    In this study, a sandwich immunoassay method utilizing enzymatic activity of alkaline phosphatase (ALP) on 5-bromo-4-chloro-3-indolyl phosphate (BCIP) for Escherichia coli (E. coli) detection was developed using surface enhanced Raman spectroscopy (SERS). For this purpose, spherical magnetic gold coated core-shell nanoparticles (MNPs-Au) and rod shape gold nanoparticles (Au-NRs) were synthesized and modified for immunomagnetic separation (IMS) of E. coli from the solution. In order to specify the developed method to ALP activity, Au-NRs were labeled with this enzyme. After successful construction of the immunoassay, BCIP substrate was added to produce the SERS-active product; 5-bromo-4-chloro-3-indole (BCI). A good linearity (R2 = 0.992) was established between the specific SERS intensity of BCI at 600 cm- 1 and logarithmic E. coli concentration in the range of 1.7 × 101-1.7 × 106 cfu mL- 1. LOD and LOQ values were also calculated and found to be 10 cfu mL- 1 and 30 cfu mL- 1, respectively.

  16. Assessment of corticosteroid-induced alkaline phosphatase as a prognostic indicator in canine lymphoma.

    PubMed

    Wiedemann, A L; Charney, S C; Barger, A M; Schaeffer, D J; Kitchell, B E

    2005-04-01

    To examine the incidence of elevated corticosteroid-induced alkaline phosphatase (sALP) in dogs with lymphoma and to determine if sALP is a reliable prognostic indicator in canine lymphoma. The medical records of 62 canine lymphoma patients treated with a combination chemotherapy protocol from 1994 to 2003 at the University of Illinois Veterinary Teaching Hospital were examined. Variables assessed with respect to response rate and remission duration included age, bodyweight, sex, breed, World Health Organization stage (I to V), substage (a or b), pretreatment administration of corticosteroid, and serum levels of alkaline phosphatase, sALP and alanine aminotransferase. sALP was not statistically significant with respect to response rate or duration of remission, nor was preinduction glucocorticoid administration. Stage was significant with respect to achieving remission. It was found that sALP is not a useful prognostic indicator for response rate and remission duration in dogs with lymphoma.

  17. Photoinduced electron transfer between Fe(III) and adenosine triphosphate-BODIPY conjugates: Application to alkaline-phosphatase-linked immunoassay.

    PubMed

    Lin, Jia-Hui; Yang, Ya-Chun; Shih, Ya-Chen; Hung, Szu-Ying; Lu, Chi-Yu; Tseng, Wei-Lung

    2016-03-15

    Fluorescent boron dipyrromethene (BODIPY) analogs are often used as sensors for detecting various species because of their relatively high extinction coefficients, outstanding fluorescence quantum yields, photostability, and pH-independent fluorescence. However, there is little-to-no information in the literature that describes the use of BODIPY analogs for detecting alkaline phosphatase (ALP) activity and inhibition. This study discovered that the fluorescence of BODIPY-conjugated adenosine triphosphate (BODIPY-ATP) was quenched by Fe(III) ions through photoinduced electron transfer. The ALP-catalyzed hydrolysis of BODIPY-ATP resulted in the formation of BODIPY-adenosine and phosphate ions. The fluorescence of the generated BODIPY-adenosine was insensitive to the change in the concentration of Fe(III) ions. Thus, the Fe(III)-induced fluorescence quenching of BODIPY-ATP can be paired with its ALP-mediated dephosphorylation to design a turn-on fluorescence probe for ALP sensing. A method detection limit at a signal-to-noise ratio of 3 for ALP was estimated to be 0.02 units/L (~6 pM; 1 ng/mL). This probe was used for the screening of ALP inhibitors, including Na3VO4, imidazole, and arginine. Because ALP is widely used in enzyme-linked immunosorbent assays, the probe was coupled to an ALP-linked immunosorbent assay for the sensitive and selective detection of immunoglobulin G (IgG). The lowest detectable concentration for IgG in this system was 5 ng/mL. Compared with the use of 3,6-fluorescein diphosphate as a signal reporter in an ALP-linked immunosorbent assay, the proposed system provided comparable sensitivity, large linear range, and high stability over temperature and pH changes. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    PubMed

    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. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Identification of rat serum alkaline phosphatase isoenzyme by means of wheat germ agglutinin.

    PubMed

    Wada, H; Niwa, N; Hayakawa, T; Tsuge, H

    1997-01-01

    Wheat germ agglutinin (WGA) precipitates bone type serum alkaline phosphatase (sALP) isoenzyme specifically. The precipitates are composed of the macromolecules of WGA and "bone type sALP" (WGA-ALP complex). In order to use bone type sALP as a marker in polyacrylamide gel electrophoresis (PAGE), a method to separate "bone type sALP" from the "WGA-ALP complex" was established by using N-acetyl-D-glucosamine (GlcNAc)-Sepharose 6E column chromatography. It was concluded that this method is useful for clinical examination in the rat.

  20. Real-time fluorescence assay of alkaline phosphatase in living cells using boron-doped graphene quantum dots as fluorophores.

    PubMed

    Chen, Li; Yang, Guancao; Wu, Ping; Cai, Chenxin

    2017-10-15

    This work reports a convenient and real-time assay of alkaline phosphatase (ALP) in living cells based on a fluorescence quench-recovery process at a physiological pH using the boron-doped graphene quantum dots (BGQDs) as fluorophore. The fluorescence of BGQDs is found to be effectively quenched by Ce 3+ ions because of the coordination of Ce 3+ ions with the carboxyl group of BGQDs. Upon addition of adenosine triphosphate (ATP) into the system, the quenched fluorescence can be recovered by the ALP-positive expressed cells (such as MCF-7 cells) due to the removal of Ce 3+ ions from BGQDs surface by phosphate ions, which are generated from ATP under catalytic hydrolysis of ALP that expressed in cells. The extent of fluorescence signal recovery depends on the level of ALP in cells, which establishes the basis of ALP assay in living cells. This approach can also be used for specific discrimination of the ALP expression levels in different type of cells and thus sensitive detection of those ALP-positive expressed cells (for example MCF-7 cells) at a very low abundance (10±5 cells mL -1 ). The advantages of this approach are that it has high sensitivity because of the significant suppression of the background due to the Ce 3+ ion quenching the fluorescence of BGQDs, and has the ability of avoiding false signals arising from the nonspecific adsorption of non-target proteins because it operates via a fluorescence quench-recovery process. In addition, it can be extended to other enzyme systems, such as ATP-related kinases. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Cloning of soluble alkaline phosphatase cDNA and molecular basis of the polymorphic nature in alkaline phosphatase isozymes of Bombyx mori midgut.

    PubMed

    Itoh, M; Kanamori, Y; Takao, M; Eguchi, M

    1999-02-01

    A cDNA coding for soluble type alkaline phosphatase (sALP) of Bombyx mori was isolated. Deduced amino acid sequence showed high identities to various ALPs and partial similarities to ATPase of Manduca sexta. Using this cDNA sequence as a probe, the molecular basis of electrophoretic polymorphism in sALP and membrane-bound type ALP (mALP) was studied. As for mALP, the result suggested that post-translational modification was important for the proteins to express activity and to represent their extensive polymorphic nature, whereas the magnitude of activities was mainly regulated by transcription. On the other hand, sALP zymogram showed poor polymorphism, but one exception was the null mutant, in which the sALP gene was largely lost. Interestingly, the sALP gene was shown to be transcribed into two mRNAs of different sizes, 2.0 and 2.4 Kb. In addition to the null mutant of sALP, we found a null mutant for mALP. Both of these mutants seem phenotypically silent, suggesting that the functional differentiation between these isozymes is not perfect, so that they can still work mutually and complement each other as an indispensable enzyme for B. mori.

  2. Synthesis and characterization of nanosized calcium phosphates by flame spray pyrolysis, and their effect on osteogenic differentiation of stem cells

    NASA Astrophysics Data System (ADS)

    Ataol, Sibel; Tezcaner, Ayşen; Duygulu, Ozgur; Keskin, Dilek; Machin, Nesrin E.

    2015-02-01

    The present study evaluates the synthesis of biocompatible osteoconductive and osteoinductive nano calcium phosphate (CaP) particles by industrially applied, aerosol-derived flame spray pyrolysis method for biomedical field. Calcium phosphate nanoparticles were produced in a range of calcium-to-phosphorus ratio, (1.20-2.19) in order to analyze the morphology and crystallinity changes, and to test the bioactivity of particles. The characterization results confirmed that nanometer-sized, spherical calcium phosphate particles were produced. The average primary particle size was determined as 23 nm by counting more than 500 particles in TEM pictures. XRD patterns, HRTEM, SAED, and SEM analyses revealed the amorphous nature of the as-prepared nano calcium phosphate particles at low Ca/P ratios. Increases in the specific surface area and crystallinity were observed with the increasing Ca/P ratio. TGA-DTA analysis showed that the thermally stable crystal phases formed after 700 °C. Cell culture studies were conducted with urine-derived stem cells that possess the characteristics of mesenchymal stem cells. Synthesized amorphous nanoparticles did not have cytotoxic effect at 5-50 μg/ml concentration range. Cells treated with the as-prepared nanoparticles had higher alkaline phosphatase (ALP) enzyme activity than control cells, indicating osteogenic differentiation of cells. A slight decrease in ALP activity of cells treated with two highest Ca:P ratios at 50 μg/ml concentration was observed at day 7. The findings suggest that calcium phosphate nanoparticles produced in this work have a potential to be used as biomaterials in biomedical applications.

  3. Comparison of Salivary pH, Buffering Capacity and Alkaline Phosphatase in Smokers and Healthy Non-Smokers

    PubMed Central

    Ahmadi-Motamayel, Fatemeh; Falsafi, Parisa; Goodarzi, Mohammad T.; Poorolajal, Jalal

    2016-01-01

    Objectives: Saliva contains alkaline phosphatase (ALP)—a key intracellular enzyme related to destructive processes and cellular damage—and has buffering capacity (BC) against acids due to the presence of bicarbonate and phosphate ions. Smoking may have deleterious effects on the oral environment due to pH changes which can affect ALP activity. This study aimed to evaluate the salivary pH, BC and ALP activity of male smokers and healthy non-smokers. Methods: This retrospective cohort study took place between August 2012 and December 2013. A total of 251 healthy male non-smokers and 259 male smokers from Hamadan, Iran, were selected. Unstimulated whole saliva was collected from each participant and pH and BC were determined using a pH meter. Salivary enzymes were measured by spectrophotometric assay. Results: Mean salivary pH (7.42 ± 0.48 and 7.52 ± 0.43, respectively; P = 0.018) and BC (3.41 ± 0.54 and 4.17 ± 0.71; P = 0.001) was significantly lower in smokers compared to non-smokers. Mean ALP levels were 49.58 ± 23.33 IU/L among smokers and 55.11 ± 27.85 IU/L among non-smokers (P = 0.015). Conclusion: Significantly lower pH, BC and ALP levels were observed among smokers in comparison to a healthy control group. These salivary alterations could potentially be utilised as biochemical markers for the evaluation of oral tissue function and side-effects among smokers. Further longitudinal studies are recommended to evaluate the effects of smoking on salivary components. PMID:27606111

  4. Assessment of the serum levels of bone alkaline phosphatase with a new immunoradiometric assay in patients with metabolic bone disease

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Garnero, P.; Delmas, P.D.

    1993-10-01

    The authors measured serum bone alkaline phosphatase (B-ALP) with a new immunoradiometric assay (IRMA) in a large sample of healthy controls comprising 173 women and 180 men, 20-88 yr of age, and in patients with metabolic bone disease. Using serum samples from patients with liver disease and patients with Paget's disease with elevated total alkaline phosphatase (T-ALP) as a source of, respectively, liver and bone isoenyzmes, they determined a liver cross-reactivity of the IRMA of 16% that was confirmed by electrophoresis of the circulating alkaline phosphatase isoenzymes. The IRMA was linear for serial sample dilutions, the recovery ranged from 89-110%,more » and the intra- and interassay variations were below 7% and 9%, respectively. B-ALP increased linearly with age in both sexes, and the mean B-ALP serum levels were not significantly different for women and men (11.3 [+-] 4.8 ng/mL for women; 11.0 [+-] 4.0 ng/mL for men). The increase in B-ALP after the menopause was significantly higher than that in T-ALP (+77% vs. +24%; P<0.001). When the values of postmenopausal women were expressed as the SD from the mean of premenopausal women, the mean Z scores were 2.2[+-] 1.8 for B-ALP and 0.9 [+-] 1.3 for T-ALP (P<0.001 between the two).« less

  5. Wnt/β-Catenin Expression Does Not Correlate with Serum Alkaline Phosphatase Concentration in Canine Osteosarcoma Patients

    PubMed Central

    Piskun, Caroline M.; Muthuswamy, Anantharaman; Huelsmeyer, Michael K.; Thompson, Victoria; Stein, Timothy J.

    2011-01-01

    Osteosarcoma is an aggressive malignancy of the bone and an increase in serum alkaline phosphatase concentration has clinical prognostic value in both humans and canines. Increased serum alkaline phosphatase concentration at the time of diagnosis has been associated with poorer outcomes for osteosarcoma patients. The biology underlying this negative prognostic factor is poorly understood. Given that activation of the Wnt signaling pathway has been associated with alkaline phosphatase expression in osteoblasts, we hypothesized that the Wnt/β-catenin signaling pathway would be differentially activated in osteosarcoma tissue based on serum ALP status. Archived canine osteosarcoma samples and primary canine osteosarcoma cell lines were used to evaluate the status of Wnt/β-catenin signaling pathway activity through immunohistochemical staining, western immunoblot analyses, quantitative reverse-transcription polymerase chain reaction, and a Wnt-responsive promoter activity assay. We found no significant difference in β-catenin expression or activation between OSA populations differing in serum ALP concentration. Pathway activity was mildly increased in the primary OSA cell line generated from a patient with increased serum ALP compared to the normal serum ALP OSA cell line. Further investigation into the mechanisms underlying differences in serum ALP concentration is necessary to improve our understanding of the biological implications of this negative prognostic indicator. PMID:22022527

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

    NASA Astrophysics Data System (ADS)

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

    2017-02-01

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

  7. Comparison of Salivary pH, Buffering Capacity and Alkaline Phosphatase in Smokers and Healthy Non-Smokers: Retrospective cohort study.

    PubMed

    Ahmadi-Motamayel, Fatemeh; Falsafi, Parisa; Goodarzi, Mohammad T; Poorolajal, Jalal

    2016-08-01

    Saliva contains alkaline phosphatase (ALP)-a key intracellular enzyme related to destructive processes and cellular damage-and has buffering capacity (BC) against acids due to the presence of bicarbonate and phosphate ions. Smoking may have deleterious effects on the oral environment due to pH changes which can affect ALP activity. This study aimed to evaluate the salivary pH, BC and ALP activity of male smokers and healthy non-smokers. This retrospective cohort study took place between August 2012 and December 2013. A total of 251 healthy male non-smokers and 259 male smokers from Hamadan, Iran, were selected. Unstimulated whole saliva was collected from each participant and pH and BC were determined using a pH meter. Salivary enzymes were measured by spectrophotometric assay. Mean salivary pH (7.42 ± 0.48 and 7.52 ± 0.43, respectively; P = 0.018) and BC (3.41 ± 0.54 and 4.17 ± 0.71; P = 0.001) was significantly lower in smokers compared to non-smokers. Mean ALP levels were 49.58 ± 23.33 IU/L among smokers and 55.11 ± 27.85 IU/L among non-smokers (P = 0.015). Significantly lower pH, BC and ALP levels were observed among smokers in comparison to a healthy control group. These salivary alterations could potentially be utilised as biochemical markers for the evaluation of oral tissue function and side-effects among smokers. Further longitudinal studies are recommended to evaluate the effects of smoking on salivary components.

  8. Cyanotoxins: a poison that frees phosphate.

    PubMed

    Raven, John A

    2010-10-12

    Autotrophic organisms obtain phosphorus from the environment by secreting alkaline phosphatases that act on esters, resulting in inorganic phosphate that is then taken up. New work shows that the cyanobacterium Aphanizomenon ovalisporum obtains inorganic phosphate by secreting the cyanotoxin cylindrospermopsin, which induces alkaline phosphatase in other phytoplankton species. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Optimization of culture conditions for osteogenically-induced mesenchymal stem cells in β-tricalcium phosphate ceramics with large interconnected channels.

    PubMed

    Bernhardt, Anne; Lode, Anja; Peters, Fabian; Gelinsky, Michael

    2011-06-01

    The aim of this study was to optimize culture conditions for human mesenchymal stem cells (hMSCs) in β-tricalcium phosphate ceramics with large interconnected channels. Fully interconnected macrochannels comprising pore diameters of 750 µm and 1400 µm were inserted into microporous β-tricalcium phosphate (β-TCP) scaffolds by milling. Human bone marrow-derived MSCs were seeded into the scaffolds and cultivated for up to 3 weeks in both static and perfusion culture in the presence of osteogenic supplements (dexamethasone, β-glycerophosphate, ascorbate). It was confirmed by scanning electron microscopic investigations and histological staining that the perfusion culture resulted in uniform distribution of cells inside the whole channel network, whereas the statically cultivated cells were primarily found at the surface of the ceramic samples. It was also determined that perfusion with standard medium containing 10% fetal calf serum (FCS) led to a strong increase (seven-fold) of cell numbers compared with static cultivation observed after 3 weeks. Perfusion with low-serum medium (2% FCS) resulted in moderate proliferation rates which were comparable to those achieved in static culture, although the specific alkaline phosphatase (ALP) activity increased by a factor of more than 3 compared to static cultivation. Gene expression analysis of the ALP gene also revealed higher levels of ALP mRNA in low-serum perfused samples compared to statically cultivated constructs. In contrast, gene expression of the late osteogenic marker bone sialoprotein II (BSPII) was decreased for perfused samples compared to statically cultivated samples. Copyright © 2010 John Wiley & Sons, Ltd.

  10. Influence of Partial O₂ Pressure on the Adhesion, Proliferation, and Osteogenic Differentiation of Human Dental Pulp Stem Cells on β-Tricalcium Phosphate Scaffold.

    PubMed

    Viña-Almunia, Jose; Mas-Bargues, Cristina; Borras, Consuelo; Gambini, Juan; El Alami, Marya; Sanz-Ros, Jorge; Peñarrocha, Miguel; Vina, Jose

    To analyze, in vitro, the influence of O₂ pressure on the adhesion, proliferation, and osteogenic differentiation of human dental pulp stem cells (DPSC) on β-tricalcium phosphate (β-TCP) scaffold. DPSC, positive for the molecular markers CD133, Oct4, Nestin, Stro-1, and CD34, and negative for CD45, were isolated from extracted third molars. Experiments were started by seeding 200,000 cells on β-TCP cultured under 3% or 21% O₂ pressure. No osteogenic medium was used. Eight different cultures were performed at each time point under each O₂ pressure condition. Cell adhesion, proliferation, and differentiation over the biomaterial were evaluated at 7, 13, 18, and 23 days of culture. Cell adhesion was determined by light microscopy, proliferation by DNA quantification, and osteogenic differentiation by alkaline phosphatase (ALP) activity analysis. DPSC adhered to β-TCP with both O₂ conditions. Cell proliferation was found from day 7 of culture. Higher values were recorded at 3% O₂ in each time point. Statistically significant differences were recorded at 23 days of culture (P = .033). ALP activity was not detectable at 7 days. There was, however, an increase in ALP activity over time in both groups. At 13, 18, and 23 days of culture, higher ALP activity was recorded under 3% O₂ pressure. Statistical differences were found at day 23 (P = .014). DPSC display capacity of adhering to β-TCP under 3% or 21% O₂ pressure conditions. Cell proliferation on β-TCP phosphate is significantly higher at 3% than at 21% O₂ pressure, the most frequently used O₂ tension. β-TCP can itself promote osteogenic differentiation of DPSC and is enhanced under 3% O₂ compared with 21%.

  11. Determination of Cancer Cell-Based pH-Sensitive Fluorescent Carbon Nanoparticles of Cross-Linked Polydopamine by Fluorescence Sensing of Alkaline Phosphatase Activity on Coated Surfaces and Aqueous Solution.

    PubMed

    Kang, Eun Bi; Choi, Cheong A; Mazrad, Zihnil Adha Islamy; Kim, Sung Han; In, Insik; Park, Sung Young

    2017-12-19

    The tumor-specific sensitive fluorescence sensing of cellular alkaline phosphatase (ALP) activity on the basis of host-guest specific and pH sensitivity was conducted on coated surfaces and aqueous states. Cross-linked fluorescent nanoparticles (C-FNP) consisting of β-cyclodextrin (β-CD)/boronic acid (BA) and fluorescent hyaluronic acid [FNP(HA)] were conjugated to fluorescent polydopamine [FNP(pDA)]. To determine the quenching effect of this system, hydrolysis of 4-nitrophenyl phosphate (NPP) to 4-nitrophenol (NP) was performed in the cavity of β-CD in the presence of ALP activated photoinduced electron transfer (PET) between NP and C-FNP. At an ALP level of 30-1000 U/L, NP caused off-emission of C-FNP because of their specific host-guest recognition. Fluorescence can be recovered under pH shock due to cleavage of the diol bond between β-CD and BA, resulting in release of NP from the fluorescent system. Sensitivity of the assays was assessed by confocal imaging not only in aqueous states, but also for the first time on coated surfaces in MDAMB-231 and MDCK cells. This novel system demonstrated high sensitivity to ALP through generation of good electron donor/acceptor pair during the PET process. Therefore, this fluorescence sensor system can be used to enhance ALP monitoring and cancer diagnosis on both coated surfaces and in aqueous states in clinical settings.

  12. Identification of human pulmonary alkaline phosphatase isoenzymes.

    PubMed

    Capelli, A; Cerutti, C G; Lusuardi, M; Donner, C F

    1997-04-01

    An increase of alkaline phosphatase (ALP) activity has been observed in the bronchoalveolar lavage fluid (BALF) of patients affected by pulmonary fibrosis in chronic interstitial lung disorders. To characterize the ALP isoenzymes in such cases, we used gel filtration, agarose gel electrophoresis, heat and amino acid inhibition assays, wheat-germ agglutinin (WGA) precipitation, and an immunoassay specific for the bone-isoform of ALP. Only one anodic band representing a high-molecular-weight isoform of ALP (Mr approximately 2,000 kDa) was observed on electrophoresis of BALF. The inhibition assay results were consistent for a tissue-nonspecific isoenzyme sensitive to a temperature of 56 degrees C (71.9 +/- 2.5% inhibition) and to homoarginine (65.7 +/- 1.9%), and resistant to L-phenylalanine and L-leucine. Less than 13% of ALP activity was heat-stable. After incubation of BALF specimens with glycosyl-phosphatidylinositol-phospholipase D plus Nonidet P-40, or with phosphatidylinositol-phospholipase C alone, an electrophoretic cathodic band (Mr approximately 220 kDa) appeared near the bone band of a standard serum. With the WGA assay, 84.4 +/- 3.3% of ALP precipitated and the band disappeared. After immunoassay for the bone isoform, a mean of less than 5% enzyme activity was measured. We conclude that the ALP found in BALF is a pulmonary isoform of a tissue nonspecific isoenzyme.

  13. Posttranslational heterogeneity of bone alkaline phosphatase in metabolic bone disease.

    PubMed

    Langlois, M R; Delanghe, J R; Kaufman, J M; De Buyzere, M L; Van Hoecke, M J; Leroux-Roels, G G

    1994-09-01

    Bone alkaline phosphatase is a marker of osteoblast activity. In order to study the posttranscriptional modification (glycosylation) of bone alkaline phosphatase in bone disease, we investigated the relationship between mass and catalytic activity of bone alkaline phosphatase in patients with osteoporosis and hyperthyroidism. Serum bone alkaline phosphatase activity was measured after lectin precipitation using the Iso-ALP test kit. Mass concentration of bone alkaline phosphatase was determined with an immunoradiometric assay (Tandem-R Ostase). In general, serum bone alkaline phosphatase mass and activity concentration correlated well. The activity : mass ratio of bone alkaline phosphatase was low in hyperthyroidism. Activation energy of the reaction catalysed by bone alkaline phosphatase was high in osteoporosis and in hyperthyroidism. Experiments with neuraminidase digestion further demonstrated that the thermodynamic heterogeneity of bone alkaline phosphatase can be explained by a different glycosylation of the enzyme.

  14. Effect of zinc phosphate chemical conversion coating on corrosion behaviour of mild steel in alkaline medium: protection of rebars in reinforced concrete.

    PubMed

    Simescu, Florica; Idrissi, Hassane

    2008-12-01

    We outline the ability of zinc phosphate coatings, obtained by chemical conversion, to protect mild steel rebars against localized corrosion, generated by chloride ions in alkaline media. The corrosion resistance of coated steel, in comparison with uncoated rebars and coated and uncoated steel rebars embedded in mortar, were evaluated by open-circuit potential, potentiodynamic polarization, cronoamperometry and electrochemical impedance spectroscopy. The coated surfaces were characterized by x-ray diffraction and scanning electron microscopy. First, coated mild steel rebars were studied in an alkaline solution with and without chloride simulating a concrete pore solution. The results showed that the slow dissolution of the coating generates hydroxyapatite Ca 10 (PO 4 ) 6 (OH) 2 . After a long immersion, the coating became dense and provided an effective corrosion resistance compared with the mild steel rebar. Secondly, the coated and uncoated steel rebars embedded in mortar and immersed in chloride solution showed no corrosion or deterioration of the coated steel. Corrosion rate is considerably lowered by this phosphate coating.

  15. Effect of zinc phosphate chemical conversion coating on corrosion behaviour of mild steel in alkaline medium: protection of rebars in reinforced concrete

    NASA Astrophysics Data System (ADS)

    Simescu, Florica; Idrissi, Hassane

    2008-12-01

    We outline the ability of zinc phosphate coatings, obtained by chemical conversion, to protect mild steel rebars against localized corrosion, generated by chloride ions in alkaline media. The corrosion resistance of coated steel, in comparison with uncoated rebars and coated and uncoated steel rebars embedded in mortar, were evaluated by open-circuit potential, potentiodynamic polarization, cronoamperometry and electrochemical impedance spectroscopy. The coated surfaces were characterized by x-ray diffraction and scanning electron microscopy. First, coated mild steel rebars were studied in an alkaline solution with and without chloride simulating a concrete pore solution. The results showed that the slow dissolution of the coating generates hydroxyapatite Ca10(PO4)6(OH)2. After a long immersion, the coating became dense and provided an effective corrosion resistance compared with the mild steel rebar. Secondly, the coated and uncoated steel rebars embedded in mortar and immersed in chloride solution showed no corrosion or deterioration of the coated steel. Corrosion rate is considerably lowered by this phosphate coating.

  16. Effect of zinc phosphate chemical conversion coating on corrosion behaviour of mild steel in alkaline medium: protection of rebars in reinforced concrete

    PubMed Central

    Simescu, Florica; Idrissi, Hassane

    2008-01-01

    We outline the ability of zinc phosphate coatings, obtained by chemical conversion, to protect mild steel rebars against localized corrosion, generated by chloride ions in alkaline media. The corrosion resistance of coated steel, in comparison with uncoated rebars and coated and uncoated steel rebars embedded in mortar, were evaluated by open-circuit potential, potentiodynamic polarization, cronoamperometry and electrochemical impedance spectroscopy. The coated surfaces were characterized by x-ray diffraction and scanning electron microscopy. First, coated mild steel rebars were studied in an alkaline solution with and without chloride simulating a concrete pore solution. The results showed that the slow dissolution of the coating generates hydroxyapatite Ca10(PO4)6(OH)2. After a long immersion, the coating became dense and provided an effective corrosion resistance compared with the mild steel rebar. Secondly, the coated and uncoated steel rebars embedded in mortar and immersed in chloride solution showed no corrosion or deterioration of the coated steel. Corrosion rate is considerably lowered by this phosphate coating. PMID:27878037

  17. Alkaline hydrolysis of ethylene phosphate: an ab initio study by supermolecule model and polarizable continuum approach.

    PubMed

    Xia, Futing; Zhu, Hua

    2011-09-01

    The alkaline hydrolysis reaction of ethylene phosphate (EP) has been investigated using a supermolecule model, in which several explicit water molecules are included. The structures and single-point energies for all of the stationary points are calculated in the gas phase and in solution at the B3LYP/6-31++G(df,p) and MP2/6-311++G(df,2p) levels. The effect of water bulk solvent is introduced by the polarizable continuum model (PCM). Water attack and hydroxide attack pathways are taken into account for the alkaline hydrolysis of EP. An associative mechanism is observed for both of the two pathways with a kinetically insignificant intermediate. The water attack pathway involves a water molecule attacking and a proton transfer from the attacking water to the hydroxide in the first step, followed by an endocyclic bond cleavage to the leaving group. While in the first step of the hydroxide attack pathway the nucleophile is the hydroxide anion. The calculated barriers in aqueous solution for the water attack and hydroxide attack pathways are all about 22 kcal/mol. The excellent agreement between the calculated and observed values demonstrates that both of the two pathways are possible for the alkaline hydrolysis of EP. Copyright © 2011 Wiley Periodicals, Inc.

  18. Blooming gelatin: an individual additive for enhancing nanoapatite precipitation, physical properties, and osteoblastic responses of nanostructured macroporous calcium phosphate bone cements.

    PubMed

    Orshesh, Ziba; Hesaraki, Saeed; Khanlarkhani, Ali

    2017-01-01

    In recent years, there has been a great interest in using natural polymers in the composition of calcium phosphate bone cements to enhance their physical, mechanical, and biological performance. Gelatin is a partially hydrolyzed form of collagen, a natural component of bone matrix. In this study, the effect of blooming gelatin on the nanohydroxyapatite precipitation, physical and mechanical properties, and cellular responses of a calcium phosphate bone cement (CPC) was investigated. Various concentrations of blooming gelatin (2, 5, and 8 wt.%) were used as the cement liquid and an equimolar mixture of tetracalcium phosphate and dicalcium phosphate was used as solid phase. The CPC without any gelatin additive was also evaluated as a control group. The results showed that gelatin accelerated hydraulic reactions of the cement paste, in which the reactants were immediately converted into nanostructured apatite precipitates after hardening. Gelatin molecules induced 4%-10% macropores (10-300 μm) into the cement structure, decreased initial setting time by ~190%, and improved mechanical strength of the as-set cement. Variation in the above-mentioned properties was influenced by the gelatin concentration and progressed with increasing the gelatin content. The numbers of the G-292 osteoblastic cells on gelatin-containing CPCs were higher than the control group at entire culture times (1-14 days), meanwhile better alkaline phosphatase (ALP) activity was determined using blooming gelatin additive. The observation of cell morphologies on the cement surfaces revealed an appropriate cell attachment with extended cell membranes on the cements. Overall, adding gelatin to the composition of CPC improved the handling characteristics such as setting time and mechanical properties, enhanced nanoapatite precipitation, and augmented the early cell proliferation rate and ALP activity.

  19. The relationship between alkaline phosphatase and bone alkaline phosphatase activity and the growth hormone/insulin-like growth factor-1 axis and vitamin D status in children with growth hormone deficiency.

    PubMed

    Witkowska-Sędek, Ewelina; Stelmaszczyk-Emmel, Anna; Majcher, Anna; Demkow, Urszula; Pyrżak, Beata

    2018-04-13

    The relationships between bone turnover, the growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis and vitamin D are complex, but still not fully explained. The GH/IGF-1 axis and vitamin D can mutually modulate each other's metabolism and influence the activation of cell proliferation, maturation, and mineralization as well as bone resorption. The aim of this study was to evaluate the reciprocal associations between bone formation markers [alkaline phosphatase (ALP), bone alkaline phosphatase (BALP)], the GH/IGF-1 axis and 25-hydroxyvitamin D [25(OH)D] in children with growth hormone deficiency at baseline and during recombinant human growth hormone (rhGH) therapy. ALP, BALP, 25(OH)D and IGF-1 levels were evaluated in 53 patients included in this prospective three-year study. ALP, BALP and IGF-1 increased during rhGH therapy. Baseline ALP activity correlated positively with baseline height velocity (HV). ALP and BALP activity at 12 months correlated positively with HV in the first year of therapy. We found positive correlations between ALP and IGF-1 at baseline and during the first year of therapy, between BALP activity at 12 months and rhGH dose in the first year of therapy, and between doses of cholecalciferol in the first year of rhGH therapy and early changes in BALP activity during rhGH therapy. Our results indicate that vitamin D supplementation enhances the effect of rhGH on bone formation process, which could improve the effects of rhGH therapy. ALP and BALP activity are useful in the early prediction of the effects of rhGH therapy, but their utility as long-term predictors seemed insufficient.

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

    PubMed

    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. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Acid and Alkaline Phosphatase Levels in GCF during Orthodontic Tooth Movement

    PubMed Central

    Farahani, Mohammad; Safavi, Seyed Mohammadreza; Dianat, Omid; Khoramian Tusi, Somayeh; Younessian, Farnaz

    2015-01-01

    Statement of the Problem The present constituents of gingival crevicular fluid (GCF) can reflect the changes occurring in underlying tissues. Considering variety of biologic bone markers, alkaline phosphatase and acid phosphatase have been examined as bone turn over markers in orthodontic tooth movement. Purpose The current study designed in a longitudinal pattern to determine the changes of acid and alkaline phosphatase (ACP & ALP) in GCF during orthodontic tooth movement. Materials and Method An upper canines from twelve patients (mean age: 14±2 years) undergoing extraction orthodontic treatment for distal movement served as the test tooth (DC), and its contralateral (CC) and antagonist (AC) canines were used as controls. The CC was included in orthodontic appliance without orthodontic force; the AC was free from any orthodontic appliance. The GCF around the experimental teeth was harvested from mesial and distal tooth sites immediately before appliance placement (T0), and 14 (T2) and 28 days (T3) after it and ALP and ACP concentration were determined spectrophotometrically. Results ALP concentration was elevated significantly in DC and CC groups at days 14 and 28 compared with the AC. In DC group, the ALP was significantly greater in mesial sites than distal site, while no significant changes were found between both sites of CC. The peak level of ALP was observed in mesial sites of DC at T2. Regarding ACP, significant elevation of this enzyme was seen in DC group both in mesial and distal sites at T2 and T3. The peak level of this enzyme was seen at T2. Conclusion Monitoring simultaneous changes of ALP and ACP levels in GCF can reflect the tissue responses occur in periodontium during bone formation and bone resorption during orthodontic tooth movement, respectively. PMID:26535403

  2. Oral antibodies to human intestinal alkaline phosphatase reduce dietary phytate phosphate bioavailability in the presence of dietary 1α-hydroxycholecalciferol.

    PubMed

    Bobeck, Elizabeth A; Hellestad, Erica M; Helvig, Christian F; Petkovich, P Martin; Cook, Mark E

    2016-03-01

    While it is well established that active vitamin D treatment increases dietary phytate phosphate utilization, the mechanism by which intestinal alkaline phosphatase (IAP) participates in phytate phosphate use is less clear. The ability of human IAP (hIAP) oral antibodies to prevent dietary phytate phosphate utilization in the presence of 1α-hydroxycholecalciferol (1α-(OH) D3) in a chick model was investigated. hIAP specific chicken immunoglobulin Y (IgY) antibodies were generated by inoculating laying hens with 17 synthetic peptides derived from the human IAP amino acid sequence and harvesting egg yolk. Western blot analysis showed all antibodies recognized hIAP and 6 of the 8 antibodies selected showed modest inhibition of hIAP activity in vitro (6 to 33% inhibition). In chicks where dietary phosphate was primarily in the form of phytate, 4 selected hIAP antibodies inhibited 1α-(OH) D3-induced increases in blood phosphate, one of which, generated against selected peptide (MFPMGTPD), was as effective as sevelamer hydrochloride in preventing the 1α-(OH) D3-induced increase in blood phosphate, but ineffective in preventing an increase in body weight gain and bone ash induced by 1α-(OH) D3. These studies demonstrated that orally-delivered antibodies to IAP limit dietary phytate-phosphate utilization in chicks treated with 1α-(OH) D3, and implicate IAP as an important host enzyme in increasing phytate phosphate bioavailability in 1α-(OH) D3 fed chicks. © 2015 Poultry Science Association Inc.

  3. Glycosylation differences contribute to distinct catalytic properties among bone alkaline phosphatase isoforms

    PubMed Central

    Linder, Cecilia Halling; Narisawa, Sonoko; Millán, José Luis; Magnusson, Per

    2009-01-01

    Three circulating human bone alkaline phosphatase (BALP) isoforms (B1, B2, and B/I) can be distinguished in healthy individuals and a fourth isoform (B1x) has been discovered in patients with chronic kidney disease and in bone tissue. The present study was designed to correlate differing glycosylation patterns of each BALP isoform with their catalytic activity towards presumptive physiological substrates and to compare those properties with two recombinant isoforms of the tissue-nonspecific ALP (TNALP) isozyme, i.e., TNALP-flag, used extensively for mutation analysis of hypophosphatasia mutations and sALP-FcD10, a chimeric enzyme recently used as therapeutic drug in a mouse model of infantile hypophosphatasia. The BALP isoforms were prepared from human osteosarcoma (SaOS-2) cells and the kinetic properties were evaluated using the synthetic substrate p-nitrophenylphosphate (pNPP) at pH 7.4 and 9.8, and the three suggested endogenous physiological substrates, i.e., inorganic pyrophosphate (PPi), pyridoxal 5′-phosphate (PLP), and phosphoethanolamine (PEA) at pH 7.4. Qualitative glycosylation differences were also assessed by lectin binding and precipitation. The kcat/KM was higher for B2 for all the investigated substrates. The catalytic activity towards PEA was essentially undetectable. The kinetic activity for TNALP-flag and sALP-FcD10 was similar to the activity of the human BALP isoforms. The BALP isoforms differed in their lectin-binding properties and dose-dependent lectin precipitation, which also demonstrated differences between native and denatured BALP isoforms. The observed differences in lectin specificity were attributed to N-linked carbohydrates. In conclusion, we demonstrate significantly different catalytic properties among the BALP isoforms due to structural differences in posttranslational glycosylation. Our data also suggests that PEA is not an endogenous substrate for the BALP isoforms or for the recombinant TNALP isoforms. The TNALP-flag and

  4. Glycosylation differences contribute to distinct catalytic properties among bone alkaline phosphatase isoforms.

    PubMed

    Halling Linder, Cecilia; Narisawa, Sonoko; Millán, José Luis; Magnusson, Per

    2009-11-01

    Three circulating human bone alkaline phosphatase (BALP) isoforms (B1, B2, and B/I) can be distinguished in healthy individuals and a fourth isoform (B1x) has been discovered in patients with chronic kidney disease and in bone tissue. The present study was designed to correlate differing glycosylation patterns of each BALP isoform with their catalytic activity towards presumptive physiological substrates and to compare those properties with two recombinant isoforms of the tissue-nonspecific ALP (TNALP) isozyme, i.e., TNALP-flag, used extensively for mutation analysis of hypophosphatasia mutations and sALP-FcD(10), a chimeric enzyme recently used as therapeutic drug in a mouse model of infantile hypophosphatasia. The BALP isoforms were prepared from human osteosarcoma (SaOS-2) cells and the kinetic properties were evaluated using the synthetic substrate p-nitrophenylphosphate (pNPP) at pH 7.4 and 9.8, and the three suggested endogenous physiological substrates, i.e., inorganic pyrophosphate (PP(i)), pyridoxal 5'-phosphate (PLP), and phosphoethanolamine (PEA) at pH 7.4. Qualitative glycosylation differences were also assessed by lectin binding and precipitation. The k(cat)/K(M) was higher for B2 for all the investigated substrates. The catalytic activity towards PEA was essentially undetectable. The kinetic activity for TNALP-flag and sALP-FcD(10) was similar to the activity of the human BALP isoforms. The BALP isoforms differed in their lectin binding properties and dose-dependent lectin precipitation, which also demonstrated differences between native and denatured BALP isoforms. The observed differences in lectin specificity were attributed to N-linked carbohydrates. In conclusion, we demonstrate significantly different catalytic properties among the BALP isoforms due to structural differences in posttranslational glycosylation. Our data also suggests that PEA is not an endogenous substrate for the BALP isoforms or for the recombinant TNALP isoforms. The TNALP

  5. Comparative evaluation of different calcium phosphate-based bone graft granules - an in vitro study with osteoblast-like cells.

    PubMed

    Bernhardt, Anne; Lode, Anja; Peters, Fabian; Gelinsky, Michael

    2013-04-01

    Granule-shaped calcium phosphate-based bone graft materials are often required for bone regeneration especially in implant dentistry. Two newly developed bone graft materials are Ceracell(®) , an open-celled highly porous bioceramic from β-tricalcium phosphate (β-TCP) under addition of bioglass and Osseolive(®) , an open porous glass ceramic with the general formula Ca2 KNa(PO4 )2 . The goal of this study was to characterize different modifications of the two bone graft materials in vitro in comparison to already established ceramic bone grafts Cerasorb M(®) , NanoBone(®) and BONIT Matrix(®) . Adhesion and proliferation of SaOS-2 osteoblast-like cells were evaluated quantitatively by determining DNA content and lactate dehydrogenase (LDH) activity and qualitatively by scanning electron microscopy (SEM). In addition, MTT cell-vitality staining was applied to confirm the attachment of viable cells to the different materials. Osteogenic differentiation was evaluated by measurement of alkaline phosphatase (ALP) activity as well as gene expression analysis of osteogenic markers using reverse transcriptase PCR. DNA content and LDH activity revealed good cell attachment and proliferation for Ceracell and Cerasorb M. When pre-incubated with cell-culture medium, also Osseolive showed good cell attachment and proliferation. Attachment and proliferation of osteoblast-like cells on NanoBone and BONIT Matrix was very low, even after pre-incubation with cell-culture medium. Specific ALP activity on Ceracell(®) , Osseolive (®) and Cerasorb M(®) increased with time and expression of bone-related genes ALP, osteonectin, osteopontin and bone sialoprotein II was demonstrated. Ceracell as well as Osseolive granules support proliferation and osteogenic differentiation in vitro and may be promising candidates for in vivo applications. © 2011 John Wiley & Sons A/S.

  6. Changes of serum alkaline phosphatase isoenzymes in fasted rats.

    PubMed

    Wada, H; Niwa, N; Hayakawa, T; Tsuge, H

    1996-10-01

    Changes of serum alkaline phosphatase (sALP) isoenzymes under fasting conditions were examined using polyacrylamide gel electrophoresis (PAGE), amino-acids (L-phenylalanine (L-Phe), L-homoarginine (L-HArg)) inhibition and wheat germ agglutinin (WGA) treatment. The sALP of non-fasted rats was separated into three bands (S1, S2, S3) by PAGE. The molecular weight (M.W.) of S1 corresponded to that of an isoenzyme found in the ileum. By the addition of L-Phe, the staining intensity of S1 was weakened, S2 and S3 remained unchanged and the total activity of the isoenzymes extracted from intestine decreased. On the other hand, the activity of isoenzymes extracted from kidney and bone decreased by the addition of L-HArg. Therefore, S1 was judged to be derived from intestine. The activities of total sALP and S1 decreased from 16 h of fasting. Total sALP activity and sALP activity of the supernatant prepared by WGA treatment decreased, whereas the ALP activity of the precipitate (difference between total sALP activity and supernatant sALP activity) did not change. The activity band of the precipitate corresponded to that of S3 by PAGE. Therefore, S3 was judged to be derived from bone. In conclusion, under fasting conditions, the activity of S1 decreased while the activities of S2 and S3 remained unchanged.

  7. iPhone-imaged and cell-powered electrophoresis titration chip for the alkaline phosphatase assay in serum by the moving reaction boundary.

    PubMed

    Cao, Xin-Yu; Kong, Fan-Zhi; Zhang, Qiang; Liu, Wei-Wen; Liu, Xiao-Ping; Li, Guo-Qing; Zhong, Ran; Fan, Liu-Yin; Xiao, Hua; Cao, Cheng-Xi

    2018-05-21

    As a vital enzyme, alkaline phosphatase (ALP) has great clinical significance in diagnoses of bone or liver cancer, bone metastases, rickets, and extrahepatic biliary obstruction. However, there is still no really portable chip for the ALP assay in blood. Herein, a simple electrophoresis titration (ET) model was developed for ALP detection via a moving reaction boundary (MRB). In the model, ALP catalyzed the dephosphorylation of a 4-methylumbelliferyl phosphate disodium salt (4-MUP) substrate in the cathode well to 4-methylumbelliferone ([4-MU]-) with a negative charge and blue fluorescence under UV excitation. After the catalysis, an electric field was used between the cathode and the anode. Under the electric field, [4-MU]- moved into the channel and neutralized the acidic Tris-HCl buffer, resulting in the quenching of [4-MU]- and creating a MRB. The ET system just had an ET chip, a lithium cell, a UV LED and an iPhone used as a recorder, having no traditional expensive power supply and fluorescence detector. The relevant method was developed, and a series of experiments were conducted via the ET chip. The experiments showed: (i) a MRB could be formed between the [4-MU]- base and the acidic buffer, and the MRB motion had a linear relationship with the ALP activity, validating the ET model; (ii) the ET run was not impacted by many interferences, implying good selectivity; and (iii) the ET chip could be used for portable detection within 10 min, implying an on-site and rapid analysis. In addition, the ET method had a relatively good sensitivity (0.1 U L-1), linearity (V = 0.033A + 3.87, R2 = 0.9980), stability (RSD 2.4-6.8%) and recoveries (101-105%). Finally, the ET method was successfully used for ALP assays in real serum samples. All the results implied that the developed method was simple, rapid and low-cost, and had potential for POCT clinical ALP assays.

  8. Bone regeneration with osteogenic matrix cell sheet and tricalcium phosphate: An experimental study in sheep.

    PubMed

    Kira, Tsutomu; Akahane, Manabu; Omokawa, Shohei; Shimizu, Takamasa; Kawate, Kenji; Onishi, Tadanobu; Tanaka, Yasuhito

    2017-10-18

    To determine the effects of a cell sheet created from sheep bone marrow and tricalcium phosphate (TCP) on osteogenesis. Bone marrow cells were harvested from a sheep and cultured in a minimal essential medium (MEM) containing ascorbic acid phosphate (AscP) and dexamethasone (Dex). After 2 wk, the formed osteogenic matrix cell sheet was lifted from the culture dish using a scraper. Additionally, harvested bone marrow cells were cultured in MEM only as a negative control group, and in MEM with AscP, Dex, and β-glycerophosphate as a positive control group. For in vitro evaluation, we measured the alkaline phosphatase (ALP) activity and osteocalcin (OC) content in the media of the cultured cells from each group. For in vivo analysis, a porous TCP ceramic was used as a scaffold. We prepared an experimental group comprising TCP scaffolds wrapped with the osteogenic matrix cell sheets and a control group consisting of the TCP scaffold only. The constructs were implanted subcutaneously into athymic rats and the cell donor sheep, and bone formation was confirmed by histology after 4 wk. In the in vitro part, the mean ALP activity was 0.39 ± 0.03 mg/well in the negative control group, 0.67 ± 0.04 mg/well in the sheet group, and 0.65 ± 0.07 mg/well in the positive control group. The mean OC levels were 1.46 ± 0.33 ng/well in the negative control group, 3.92 ± 0.16 ng/well in the sheet group, and 4.4 ± 0.47 ng/well in the positive control group, respectively. The ALP activity and OC levels were significantly higher in the cell sheet and positive control groups than in the negative control group ( P < 0.05). There was no significant difference in ALP activity or OC levels between the cell sheet group and the positive control group ( P > 0.05). TCP constructs wrapped with cell sheets prior to implantation showed bone formation, in contrast to TCP scaffolds alone, which exhibited poor bone formation when implanted, in the subcutaneous layer both in athymic rats and in the

  9. Blooming gelatin: an individual additive for enhancing nanoapatite precipitation, physical properties, and osteoblastic responses of nanostructured macroporous calcium phosphate bone cements

    PubMed Central

    Orshesh, Ziba; Hesaraki, Saeed; Khanlarkhani, Ali

    2017-01-01

    In recent years, there has been a great interest in using natural polymers in the composition of calcium phosphate bone cements to enhance their physical, mechanical, and biological performance. Gelatin is a partially hydrolyzed form of collagen, a natural component of bone matrix. In this study, the effect of blooming gelatin on the nanohydroxyapatite precipitation, physical and mechanical properties, and cellular responses of a calcium phosphate bone cement (CPC) was investigated. Various concentrations of blooming gelatin (2, 5, and 8 wt.%) were used as the cement liquid and an equimolar mixture of tetracalcium phosphate and dicalcium phosphate was used as solid phase. The CPC without any gelatin additive was also evaluated as a control group. The results showed that gelatin accelerated hydraulic reactions of the cement paste, in which the reactants were immediately converted into nanostructured apatite precipitates after hardening. Gelatin molecules induced 4%–10% macropores (10–300 μm) into the cement structure, decreased initial setting time by ~190%, and improved mechanical strength of the as-set cement. Variation in the above-mentioned properties was influenced by the gelatin concentration and progressed with increasing the gelatin content. The numbers of the G-292 osteoblastic cells on gelatin-containing CPCs were higher than the control group at entire culture times (1–14 days), meanwhile better alkaline phosphatase (ALP) activity was determined using blooming gelatin additive. The observation of cell morphologies on the cement surfaces revealed an appropriate cell attachment with extended cell membranes on the cements. Overall, adding gelatin to the composition of CPC improved the handling characteristics such as setting time and mechanical properties, enhanced nanoapatite precipitation, and augmented the early cell proliferation rate and ALP activity. PMID:28176961

  10. Reductive dechlorination of carbon tetrachloride using buffered alkaline ascorbic acid.

    PubMed

    Lin, Ya-Ting; Liang, Chenju

    2015-10-01

    Alkaline ascorbic acid (AA) was recently discovered as a novel in-situ chemical reduction (ISCR) reagent for remediating chlorinated solvents in the subsurface. For this ISCR process, the maintenance of an alkaline pH is essential. This study investigated the possibility of the reduction of carbon tetrachloride (CT) using alkaline AA solution buffered by phosphate and by NaOH. The results indicated that CT was reduced by AA, and chloroform (CF) was a major byproduct at a phosphate buffered pH of 12. However, CT was completely reduced by AA in 2M NaOH without CF formation. In the presence of iron/soil minerals, iron could be reduced by AA and Fe(2+) tends to precipitate on the mineral surface to accelerate CT degradation. A simultaneous transfer of hydrogenolysis and dichloroelimination would occur under phosphate buffered pH 12. This implies that a high alkaline environment is a crucial factor for maintaining the dominant pathway of two electron transfer from dianionic AA to dehydroascorbic acid, and to undergo dichloroelimination of CT. Moreover, threonic acid and oxalic acid were identified to be the major AA decomposition products in alkaline solutions. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

    PubMed Central

    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

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

    PubMed

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

    2016-01-01

    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. 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. 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. The results indicated that intercropping with potato onion promoted the growth and P uptake of tomato in phosphorus-rich soil and

  13. Microporous nanofibrous fibrin-based scaffolds for craniofacial bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Osathanon, Thanaphum

    The fibrotic response of the body to synthetic polymers limits their success in tissue engineering and other applications. Though porous polymers have demonstrated improved healing, difficulty in controlling their pore sizes and pore interconnections has clouded the understanding of this phenomenon. In this study, a novel method to fabricate natural polymer/calcium phosphate composite scaffolds and immobilized alkaline phosphatase fibrin scaffolds with tightly controllable pore size, pore interconnection has been investigated. Microporous, nanofibrous fibrin scaffolds (FS) were fabricated using sphere-templating method. Calcium phosphate/fibrin composite scaffolds were created by solution deposition of calcium phosphate on fibrin surfaces or by direct incorporation of nanocrystalline hydroxyapatite (nHA). The SEM results showed that fibrin scaffolds exhibited a highly porous and interconnected structure. Osteoblast-like cells, obtained from murine calvaria, attached, spread and showed a polygonal morphology on the surface of the biomaterial. Multiple cell layers and fibrillar matrix deposition were observed. Moreover, cells seeded on mineralized fibrin scaffolds (MFS) exhibited significantly higher alkaline phosphatase activity as well as osteoblast marker gene expression compared to FS and nHA incorporated fibrin scaffolds (nHA/FS). These fibrin-based scaffolds were degraded both in vitro and in vivo. Furthermore, these scaffolds promoted bone formation in a mouse calvarial defect model and the bone formation was enhanced by addition of rhBMP-2. The second approach was to immobilize alkaline phosphatase (ALP) on fibrin scaffolds. ALP enzyme was covalently immobilized on the microporous nanofibrous fibrin scaffolds using 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDC). The SEM results demonstrated mineral deposition on immobilized ALP fibrin scaffolds (ALP/FS) when incubated in medium supplemented with beta-glycerophosphate, suggesting that the

  14. Segment-specific expression of alkaline phosphatase in the Tubifex embryo requires DNA replication and mRNA synthesis.

    PubMed

    Kitamura, Kaoru; Shimizu, Takashi

    2002-04-15

    During embryogenesis of the oligochaete annelid Tubifex, segments VII and VIII specifically express mesodermal alkaline phosphatase (ALP) activity in the ventrolateral region. In this study, using specific inhibitors, we examined whether this segment-specific expression of ALP activity depends on DNA replication and RNA transcription. BrdU-incorporation experiments showed that presumptive ALP-expressing cells undergo the last round of DNA replication at 12-24 hr prior to emergence of ALP activity. When this DNA replication was inhibited by aphidicolin, ALP development was completely abrogated in the ventrolateral mesoderm. Similar inhibition of ALP development was also observed in alpha-amanitin-injected embryos. While injection of alpha-amanitin at 24 hr prior to the emergence of ALP activity exerted inhibitory effects on ALP development, injection at 14 hr was no longer effective. In contrast, ALP activity developed normally in cytochalasin-D-treated embryos in which cytokinesis was prevented from occurring for 36 hs prior to appearance of ALP activity. These results suggest that the segment-specific development of ALP activity in the Tubifex embryo depends on DNA replication and mRNA transcription, both of which occur long before the emergence of ALP activity. Copyright 2002 Wiley-Liss, Inc.

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

    NASA Astrophysics Data System (ADS)

    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.

  16. Aluminum phosphate microcapsule flame retardants for flexible polyurethane foams

    NASA Astrophysics Data System (ADS)

    Zhang, Bin; Liu, Hong; Han, Jian

    2018-04-01

    In this study, highly efficient flame-retardant aluminum phosphate (ALP) microcapsules were synthesized from ALP and ammonium phosphomolybdate trihydrate. The chemical structure of the ALP microcapsules was characterized by scanning electron microscopy and elemental analysis, and the thermal degradation behavior was investigated by thermogravimetric analysis (TGA). Subsequently, flexible polyurethane (PU) foams were prepared with the ALP microcapsules. Limiting oxygen index (LOI) tests, vertical burning tests, smoke density rating (SDR), and cone calorimetric tests were employed to investigate the combustion of the materials. The results showed that the flexible PU foams with 15 parts per hundred polyol by weight (pphp) ALP microcapsules passed the vertical burning test and they had an increased LOI value of 28.5%. The SDR value for PU/20 pphp ALP microcapsule composites was about 16.0% and the SDR value for the pure PU was about 29.0%. The corresponding flame-retardant mechanism was investigated by Fourier transform infrared spectroscopy, TGA, Pyrolysis Gas Chromatography Mass Spectrometry (Py-GC/MS) tests, and energy-dispersive X-ray spectrometry.

  17. Determination of trace alkaline phosphatase by solid-substrate room-temperature phosphorimetry based on Triticum vulgare lectin labeled with fullerenol.

    PubMed

    Liu, Jia-Ming; Gao, Fei; Huang, Hong-Hua; Zeng, Li-Qing; Huang, Xiao-Mei; Zhu, Guo-Hui; Li, Zhi-Ming

    2008-04-01

    Fullerenol (F) shows a strong and stable room-temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM) at lambda ex max/ lambda em max =542.0/709.4 nm. When modified by dodecylbenzenesulfonic acid sodium salt (DBS), fullerenol emits a stronger signal. It was also found that quantitative specific affinity-adsorption reaction can be carried out between Triticum vulgare lectin (WGA) labeled with DBS-F and alkaline phosphatase (ALP) on the surface of NCM, and the product obtained (WGA-ALP-WGA-F-DBS) emits a strong and stable RTP signal. Furthermore, the content of ALP was proportional to the DeltaI(p) value. Based on the facts above, a new method for the determination of trace amounts of ALP by affinity-adsorption solid-substrate room-temperature phosphorimetry (AA-SS-RTP) was established, using fullerenol modified with DBS to label WGA. The detection limit was 0.011 fg spot(-1) (corresponding concentration: 2.8x10(-14) g ml(-1), namely 2.8x10(-16) mol l(-1)). This method with high sensitivity, accuracy, and precision has been successfully applied to the determination of the content of ALP in human serum survey and forecast human disease, and the results are tallied with those using alkaline phosphatase kits. The mechanism for the determination of ALP using AA-SS-RTP was also discussed.

  18. Quenching of graphene quantum dots fluorescence by alkaline phosphatase activity in the presence of hydroquinone diphosphate.

    PubMed

    Pereira da Silva Neves, Marta Maria; González-García, María Begoña; Pérez-Junquera, Alejandro; Hernández-Santos, David; Fanjul-Bolado, Pablo

    2018-05-01

    In this work, a turn-off photoluminescent sensing proof-of-concept based on blue luminescent graphene quantum dots (GQDs) as the fluorescent probe was developed. For that purpose, GQDs optical response was related with the catalytic enzymatic activity of alkaline phosphatase (ALP), in the presence of hydroquinone diphosphate (HQDP). The hydrolysis of HQDP by ALP generated hydroquinone (HQ). The oxidation of HQ, enzymatically produced, to p-benzoquinone (BQ) resulted in the quenching of GQDs fluorescence (FL). Therefore, the developed luminescent sensing mechanism allowed the FL quenching with ALP activity to be related and thus quantified the concentration of ALP down to 0.5 nM of enzyme. This innovative design principle appears as a promising tool for the development of enzymatic sensors based on ALP labeling with fluorescent detection or even for direct ALP luminescent quantification in an easy, fast and sensitive manner. Copyright © 2018 John Wiley & Sons, Ltd.

  19. Evolution of the magnesium incorporated amorphous calcium phosphate to nano-crystallized hydroxyapatite in alkaline solution

    NASA Astrophysics Data System (ADS)

    Zhang, Xiao-Juan; Lin, Dong-Yang; Yan, Xiao-Hui; Wang, Xiao-Xiang

    2011-12-01

    A homogeneous amorphous calcium phosphate (ACP) coating containing magnesium was achieved on titanium substrates by electrochemical deposition (ECD). Its amorphous structure is confirmed by transmission electron microscope (TEM) together with grazing reflection absorption infrared spectroscopy (IR) spectrometer. In the images of high-resolution transmission electron microscope (HRTEM), the ACP spheres are assembled by nano-particles with the diameter of 5-10 nm. In the alkaline environment, nucleation of hydroxyapatite (HAP) occurs on the surfaces of ACP spheres. By consuming the Ca and PO 4 ions inside the ACP spheres, the HAP nuclei grow outward. Confirmed by TEM, the ACP spheres converse to hollow HAP spheres composed of HAP nano-needles. The coating is finally constructed by the HAP nano-needles, which are themselves aggregated by numerous nano-particles.

  20. Soil mineral alters the effect of Cd on the alkaline phosphatase activity.

    PubMed

    Tan, Xiangping; He, Yike; Wang, Ziquan; Li, Chenghui; Kong, Long; Tian, Haixia; Shen, Weijun; Megharaj, Mallavarapu; He, Wenxiang

    2018-05-30

    The toxicity of heavy metals (HMs) to soil enzymes is directly influenced by the status of the enzyme (free vs. immobilized on minerals) and the duration of exposure. However, little information is available on the interaction effect of HMs, mineral, and exposure time on soil enzyme activities. We investigated the interaction mechanism of alkaline phosphatase (ALP) with minerals (montmorillonite and goethite) and the response of free and immobilized ALP to cadmium (Cd) toxicity under different exposure times. The adsorption isotherms of ALP on both minerals were L-type. The maximum adsorption capacity of goethite for ALP was 3.96 times than montmorillonite, although both had similar adsorption constant (K). Goethite showed a greater inhibitory effect on ALP activity than montmorillonite. The toxicity of Cd to free- and goethite-ALP was enhanced with increasing exposure time, indicating a time-dependent inhibition. However, Cd toxicity to montmorillonite-ALP was not affected by the exposure time. The inhibition of Cd to soil enzyme activity is influenced by the properties of mineral complexes and the duration of exposure. A further understanding of the time pattern of HMs toxicity is helpful for accurately assessing the hazards of HMs to soil enzyme activity. Copyright © 2018 Elsevier Inc. All rights reserved.

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

    PubMed

    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. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Treatment of Salmonella enterica serovar Enteritidis with a sublethal concentration of trisodium phosphate or alkaline pH induces thermotolerance.

    PubMed

    Sampathkumar, Balamurugan; Khachatourians, George G; Korber, Darren R

    2004-08-01

    The responses of Salmonella enterica serovar Enteritidis to a sublethal dose of trisodium phosphate (TSP) and its equivalent alkaline pH made with NaOH were examined. Pretreatment of S. enterica serovar Enteritidis cells with 1.5% TSP or pH 10.0 solutions resulted in a significant increase in thermotolerance, resistance to 2.5% TSP, resistance to high pH, and sensitivity to acid and H(2)O(2). Protein inhibition studies with chloramphenicol revealed that thermotolerance, unlike resistance to high pH, was dependent on de novo protein synthesis. Two-dimensional polyacrylamide gel electrophoresis (PAGE) of total cellular proteins from untreated control cells resolved as many as 232 proteins, of which 22 and 15% were absent in TSP- or alkaline pH-pretreated cells, respectively. More than 50% of the proteins that were either up- or down-regulated by TSP pretreatment were also up- or down-regulated by alkaline pH pretreatment. Sodium dodecyl sulfate-PAGE analysis of detergent-insoluble outer membrane proteins revealed the up-regulation of at least four proteins. Mass spectrometric analysis showed the up-regulated proteins to include those involved in the transport of small hydrophilic molecules across the cytoplasmic membrane and those that act as chaperones and aid in the export of newly synthesized proteins by keeping them in open conformation. Other up-regulated proteins included common housekeeping proteins like those involved in amino acid biosynthesis, nucleotide metabolism, and aminoacyl-tRNA biosynthesis. In addition to the differential expression of proteins following TSP or alkaline pH treatment, changes in membrane fatty acid composition were also observed. Alkaline pH- or TSP-pretreated cells showed a higher saturated and cyclic to unsaturated fatty acid ratio than did the untreated control cells. These results suggest that the cytoplasmic membrane could play a significant role in the induction of thermotolerance and resistance to other stresses following TSP

  3. Correction of hypophosphatasia (HPP) associated mineralization deficiencies in vitro by phosphate/pyrophosphate modulation in periodontal ligament cells

    PubMed Central

    Rodrigues, Thaisângela L.; Foster, Brian L.; Silverio, Karina G.; Martins, Luciane; Casati, Marcio Z.; Sallum, Enilson A.; Somerman, Martha J.; Nociti, Francisco H.

    2013-01-01

    Background Mutations in the Alpl gene in hypophosphatasia (HPP) reduce the function of tissue nonspecific alkaline phosphatase (TNAP), resulting in increased pyrophosphate (PPi) and a severe deficiency in acellular cementum. We hypothesized that exogenous phosphate (Pi) would rescue the in vitro mineralization capacity of periodontal ligament (PDL) cells harvested from HPP-diagnosed subjects, by correcting Pi/PPi ratio and modulating expression of genes involved with Pi/PPi metabolism. Methods Ex vivo and in vitro analyses were employed to identify mechanisms involved in HPP-associated PDL/tooth root deficiencies. Constitutive expression of PPi-associated genes was contrasted in PDL versus pulp tissues obtained from healthy subjects. Primary PDL cell cultures from HPP subjects (monozygotic twin males) were established to assay alkaline phosphatase activity (ALP), in vitro mineralization, and gene expression. Exogenous Pi was provided to correct Pi/PPi ratio. Results PDL tissues obtained from healthy individuals featured higher basal expression of key PPi regulators, genes Alpl, progressive ankylosis protein (Ankh) and ectonucleotide pyrophosphatase/phosphodiesterase 1 (Enpp1), versus paired pulp tissues. A novel Alpl mutation was identified in the twin HPP subjects enrolled in this study. Compared to controls, HPP-PDL cells exhibited significantly reduced ALP and mineralizing capacity, which were rescued by addition of 1mM Pi. Dysregulated expression of PPi regulatory genes Alpl, Ankh, and Enpp1 was also corrected by adding Pi, though other matrix markers evaluated in our study remained down-regulated. Conclusions These findings underscore the importance of controlling Pi/PPi ratio toward development of a functional periodontal apparatus, and support Pi/PPi imbalance as the etiology of HPP-associated cementum defects. PMID:22014174

  4. Alkaline phosphatase activity in human colostrum as a valuable predictive biomarker for lactational mastitis in nursing mothers.

    PubMed

    Bjelakovic, Ljiljana; Kocic, Gordana; Bjelakovic, Bojko; Zivkovic, Nikola; Stojanović, Dusica; Sokolovic, Danka; Mladenovic-Ciric, Ivana; Sokolovic, Dusan

    2012-08-01

    Biochemical investigations have shown that an indigenous milk enzyme - alkaline phosphatase (ALP) - which is detectable in the lactocytes, plays a very important diagnostic role in clinical medicine, since its activity varies in different tissues and serves as a specific indicator of disease states. The purpose of this study was to evaluate ALP activity in human colostrum as a possible early predictive biomarker for lactational mastitis in nursing mothers. During a period from May to July 2010, a total of 60 healthy nursing mothers were recruited for this study. The mean level of colostrum ALP activity from the affected breasts was significantly higher when compared with ALP activity from the contralateral asymptomatic as well as 'healthy' breasts (p < 0.01). Determining ALP activity in colostrum could be a valuable biochemical marker for an early prediction of mastitis in nursing mothers.

  5. Age-related Changes in the Alkaline Phosphatase Activity of Healthy and Inflamed Human Dental Pulp.

    PubMed

    Aslantas, Eda E; Buzoglu, Hatice Dogan; Karapinar, Senem Pinar; Cehreli, Zafer C; Muftuoglu, Sevda; Atilla, Pergin; Aksoy, Yasemin

    2016-01-01

    Alkaline phosphatase (ALP) plays an important role in inducing mineralization events in the dental pulp. This study investigated and compared the ALP levels in healthy and inflamed pulp in young and old human pulp. Tissue samples were collected from young (<30 years) and old (>60 years) donors. In both age groups, healthy human pulp (n = 18) were collected from extracted wisdom teeth. For reversible and irreversible pulpitis, pulp samples (n = 18 each) were obtained during endodontic treatment. ALP activity was assessed by spectrophotometry and immunhistochemistry. Regardless of age, reversible pulpitis group samples showed a slight elevation in ALP activity compared with normal healthy pulp. In elderly patients, ALP expression with irreversible pulpitis was significantly higher than those with a healthy pulp (P < .05). In the hyperemic state, both the young and old pulp shows a slight increase in ALP activity, whereas in irreversible pulpitis, only the old pulp shows significantly elevated ALP levels. Such an increase may trigger calcification events, which may eventually cause difficulties in endodontic treatment procedures in elderly individuals. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  6. Effects of 3D-Printed Polycaprolactone/β-Tricalcium Phosphate Membranes on Guided Bone Regeneration

    PubMed Central

    Shim, Jin-Hyung; Won, Joo-Yun; Park, Jung-Hyung; Bae, Ji-Hyeon; Ahn, Geunseon; Kim, Chang-Hwan; Lim, Dong-Hyuk; Cho, Dong-Woo; Yun, Won-Soo; Bae, Eun-Bin; Jeong, Chang-Mo; Huh, Jung-Bo

    2017-01-01

    This study was conducted to compare 3D-printed polycaprolactone (PCL) and polycaprolactone/β-tricalcium phosphate (PCL/β-TCP) membranes with a conventional commercial collagen membrane in terms of their abilities to facilitate guided bone regeneration (GBR). Fabricated membranes were tested for dry and wet mechanical properties. Fibroblasts and preosteoblasts were seeded into the membranes and rates and patterns of proliferation were analyzed using a kit-8 assay and by scanning electron microscopy. Osteogenic differentiation was verified by alizarin red S and alkaline phosphatase (ALP) staining. An in vivo experiment was performed using an alveolar bone defect beagle model, in which defects in three dogs were covered with different membranes. CT and histological analyses at eight weeks after surgery revealed that 3D-printed PCL/β-TCP membranes were more effective than 3D-printed PCL, and substantially better than conventional collagen membranes in terms of biocompatibility and bone regeneration and, thus, at facilitating GBR. PMID:28441338

  7. Effects of 3D-Printed Polycaprolactone/β-Tricalcium Phosphate Membranes on Guided Bone Regeneration.

    PubMed

    Shim, Jin-Hyung; Won, Joo-Yun; Park, Jung-Hyung; Bae, Ji-Hyeon; Ahn, Geunseon; Kim, Chang-Hwan; Lim, Dong-Hyuk; Cho, Dong-Woo; Yun, Won-Soo; Bae, Eun-Bin; Jeong, Chang-Mo; Huh, Jung-Bo

    2017-04-25

    This study was conducted to compare 3D-printed polycaprolactone (PCL) and polycaprolactone/β-tricalcium phosphate (PCL/β-TCP) membranes with a conventional commercial collagen membrane in terms of their abilities to facilitate guided bone regeneration (GBR). Fabricated membranes were tested for dry and wet mechanical properties. Fibroblasts and preosteoblasts were seeded into the membranes and rates and patterns of proliferation were analyzed using a kit-8 assay and by scanning electron microscopy. Osteogenic differentiation was verified by alizarin red S and alkaline phosphatase (ALP) staining. An in vivo experiment was performed using an alveolar bone defect beagle model, in which defects in three dogs were covered with different membranes. CT and histological analyses at eight weeks after surgery revealed that 3D-printed PCL/β-TCP membranes were more effective than 3D-printed PCL, and substantially better than conventional collagen membranes in terms of biocompatibility and bone regeneration and, thus, at facilitating GBR.

  8. Effect of surface modification of nanofibres with glutamic acid peptide on calcium phosphate nucleation and osteogenic differentiation of marrow stromal cells.

    PubMed

    Karaman, Ozan; Kumar, Ankur; Moeinzadeh, Seyedsina; He, Xuezhong; Cui, Tong; Jabbari, Esmaiel

    2016-02-01

    Biomineralization is mediated by extracellular matrix (ECM) proteins with amino acid sequences rich in glutamic acid. The objective of this study was to investigate the effect of calcium phosphate deposition on aligned nanofibres surface-modified with a glutamic acid peptide on osteogenic differentiation of rat marrow stromal cells. Blend of EEGGC peptide (GLU) conjugated low molecular weight polylactide (PLA) and high molecular weight poly(lactide-co-glycolide) (PLGA) was electrospun to form aligned nanofibres (GLU-NF). The GLU-NF microsheets were incubated in a modified simulated body fluid for nucleation of calcium phosphate crystals on the fibre surface. To achieve a high calcium phosphate to fibre ratio, a layer-by-layer approach was used to improve diffusion of calcium and phosphate ions inside the microsheets. Based on dissipative particle dynamics simulation of PLGA/PLA-GLU fibres, > 80% of GLU peptide was localized to the fibre surface. Calcium phosphate to fibre ratios as high as 200%, between those of cancellous (160%) and cortical (310%) bone, was obtained with the layer-by-layer approach. The extent of osteogenic differentiation and mineralization of marrow stromal cells seeded on GLU-NF microsheets was directly related to the amount of calcium phosphate deposition on the fibres prior to cell seeding. Expression of osteogenic markers osteopontin, alkaline phosphatase (ALP), osteocalcin and type 1 collagen increased gradually with calcium phosphate deposition on GLU-NF microsheets. Results demonstrate that surface modification of aligned synthetic nanofibres with EEGGC peptide dramatically affects nucleation and growth of calcium phosphate crystals on the fibres leading to increased osteogenic differentiation of marrow stromal cells and mineralization. Copyright © 2013 John Wiley & Sons, Ltd.

  9. High phosphate feeding promotes mineral and bone abnormalities in mice with chronic kidney disease.

    PubMed

    Lau, Wei Ling; Linnes, Michael; Chu, Emily Y; Foster, Brian L; Bartley, Bryan A; Somerman, Martha J; Giachelli, Cecilia M

    2013-01-01

    Chronic kidney disease-mineral bone disorder (CKD-MBD) is a systemic syndrome characterized by imbalances in mineral homeostasis, renal osteodystrophy (ROD) and ectopic calcification. The mechanisms underlying this syndrome in individuals with chronic kidney disease (CKD) are not yet clear. We examined the effect of normal phosphate (NP) or high phosphate (HP) feeding in the setting of CKD on bone pathology, serum biochemistry and vascular calcification in calcification-prone dilute brown non-agouti (DBA/2) mice. In both NP and HP-fed CKD mice, elevated serum parathyroid hormone and alkaline phosphatase (ALP) levels were observed, but serum phosphorus levels were equivalent compared with sham controls. CKD mice on NP diet showed trabecular alterations in the long bone consistent with high-turnover ROD, including increased trabecular number with abundant osteoblasts and osteoclasts. Despite trabecular bone and serum biochemical changes, CKD/NP mice did not develop vascular calcification. In contrast, CKD/HP mice developed arterial medial calcification (AMC), more severe trabecular bone alterations and cortical bone abnormalities that included decreased cortical thickness and density, and increased cortical porosity. Cortical bone porosity and trabecular number strongly correlated with the degree of aortic calcification. HP feeding was required to induce the full spectrum of CKD-MBD symptoms in CKD mice.

  10. Canine osteosarcoma cell lines from patients with differing serum alkaline phosphatase concentrations display no behavioral differences in vitro

    PubMed Central

    Holmes, Katie E.; Thompson, Victoria; Piskun, Caroline M.; Kohnken, Rebecca A.; Huelsmeyer, Michael K.; Fan, Timothy M.; Stein, Timothy J.

    2013-01-01

    Osteosarcoma is an aggressive malignancy and represents the most frequent primary bone malignancy of dogs and humans. Prognostic factors reported for osteosarcoma include tumor size, presence of metastatic disease, and serum alkaline phosphatase (ALP) concentration at the time of diagnosis. To date, there have been no studies to determine whether the behavior of osteosarcoma cells differ based on serum ALP concentration. Here we report on the generation of six canine osteosarcoma cell lines from osteosarcoma-bearing dogs with differences in serum ALP concentration. To determine whether in vitro behavior differs between primary osteosarcoma cell lines generated from patients with normal or increased serum ALP assays were performed to evaluate proliferation, migration, invasion, and chemosensitivity. There were no significant differences in cell proliferation, migration, invasion, or chemosensitivity between cell lines associated normal or increased serum ALP concentration. PMID:23489774

  11. Canine osteosarcoma cell lines from patients with differing serum alkaline phosphatase concentrations display no behavioural differences in vitro.

    PubMed

    Holmes, K E; Thompson, V; Piskun, C M; Kohnken, R A; Huelsmeyer, M K; Fan, T M; Stein, T J

    2015-09-01

    Osteosarcoma is an aggressive malignancy and represents the most frequent primary bone malignancy of dogs and humans. Prognostic factors reported for osteosarcoma include tumour size, presence of metastatic disease and serum alkaline phosphatase (ALP) concentration at the time of diagnosis. To date, there have been no studies to determine whether the behaviour of osteosarcoma cells differ based on serum ALP concentration. Here, we report on the generation of six canine osteosarcoma cell lines from osteosarcoma-bearing dogs with differences in serum ALP concentration. To determine whether in vitro behaviour differs between primary osteosarcoma cell lines generated from patients with normal or increased serum ALP, assays were performed to evaluate proliferation, migration, invasion and chemosensitivity. There were no significant differences in cell proliferation, migration, invasion or chemosensitivity between cell lines associated with normal or increased serum ALP concentration. © 2013 Blackwell Publishing Ltd.

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

    PubMed

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

    2014-04-01

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

  13. Long-term nitrogen fertilization decreased the abundance of inorganic phosphate solubilizing bacteria in an alkaline soil

    NASA Astrophysics Data System (ADS)

    Zheng, Bang-Xiao; Hao, Xiu-Li; Ding, Kai; Zhou, Guo-Wei; Chen, Qing-Lin; Zhang, Jia-Bao; Zhu, Yong-Guan

    2017-02-01

    Inorganic phosphate solubilizing bacteria (iPSB) are essential to facilitate phosphorus (P) mobilization in alkaline soil, however, the phylogenetic structure of iPSB communities remains poorly characterized. Thus, we use a reference iPSB database to analyze the distribution of iPSB communities based on 16S rRNA gene illumina sequencing. Additionally, a noval pqqC primer was developed to quantify iPSB abundance. In our study, an alkaline soil with 27-year fertilization treatment was selected. The percentage of iPSB was 1.10~2.87% per sample, and the dominant iPSB genera were closely related to Arthrobacter, Bacillus, Brevibacterium and Streptomyces. Long-term P fertilization had no significant effect on the abundance of iPSB communities. Rather than P and potassium (K) additions, long-term nitrogen (N) fertilization decreased the iPSB abundance, which was validated by reduced relative abundance of pqqC gene (pqqC/16S). The decreased iPSB abundance was strongly related to pH decline and total N increase, revealing that the long-term N additions may cause pH decline and subsequent P releases relatively decreasing the demands of the iPSB community. The methodology and understanding obtained here provides insights into the ecology of inorganic P solubilizers and how to manipulate for better P use efficiency.

  14. Genome-wide association study on serum alkaline phosphatase levels in a Chinese population

    PubMed Central

    2013-01-01

    Background Serum alkaline phosphatase (ALP) is a complex phenotype influenced by both genetic and environmental factors. Recent Genome-Wide Association Studies (GWAS) have identified several loci affecting ALP levels; however, such studies in Chinese populations are limited. We performed a GWAS analyzing the association between 658,288 autosomal SNPs and serum ALP in 1,461 subjects, and replicated the top SNPs in an additional 8,830 healthy Chinese Han individuals. The interactions between significant locus and environmental factors on serum ALP levels were further investigated. Results The association between ABO locus and serum ALP levels was replicated (P = 2.50 × 10-21, 1.12 × 10-56 and 2.82 × 10-27 for SNP rs8176720, rs651007 and rs7025162 on ABO locus, respectively). SNP rs651007 accounted for 2.15% of the total variance of serum ALP levels independently of the other 2 SNPs. When comparing our findings with previously published studies, ethnic differences were observed across populations. A significant interaction between ABO rs651007 and overweight and obesity was observed (FDR for interaction was 0.036); for individuals with GG genotype, those with normal weight and those who were overweight or obese have similar serum ALP concentrations; minor allele A of rs651007 remarkably reduced serum ALP levels, but this effect was attenuated in overweight and obese individuals. Conclusions Our findings indicate that ABO locus is a major determinant for serum ALP levels in Chinese Han population. Overweight and obesity modifies the effect of ABO locus on serum ALP concentrations. PMID:24094242

  15. A segmental pattern of alkaline phosphatase activity within the notochord coincides with the initial formation of the vertebral bodies.

    PubMed

    Grotmol, Sindre; Nordvik, Kari; Kryvi, Harald; Totland, Geir K

    2005-05-01

    This study shows that segmental expression of alkaline phosphatase (ALP) activity by the notochord of the Atlantic salmon (Salmo salar L.) coincides with the initial mineralization of the vertebral body (chordacentrum), and precedes ALP expression by presumed somite-derived cells external to the notochordal sheath. The early expression of ALP indicates that the notochord plays an instructive role in the segmental patterning of the vertebral column. The chordacentra form segmentally as mineralized rings within the notochordal sheath, and ALP activity spreads within the chordoblast layer from ventral to dorsal, displaying the same progression and spatial distribution as the mineralization process. No ALP activity was observed in sclerotomal mesenchyme surrounding the notochordal sheath during initial formation of the chordacentra. Our results support previous findings indicating that the chordoblasts initiate a segmental differentiation of the notochordal sheath into chordacentra and intervertebral regions.

  16. Associations between serum bone-specific alkaline phosphatase activity, biochemical parameters, and functional polymorphisms of the tissue-nonspecific alkaline phosphatase gene in a Japanese population.

    PubMed

    Sogabe, Natsuko; Tanabe, Rieko; Haraikawa, Mayu; Maruoka, Yutaka; Orimo, Hideo; Hosoi, Takayuki; Goseki-Sone, Masae

    2013-01-01

    We had demonstrated that single nucleotide polymorphism (787T>C) in the tissue-nonspecific ALP (TNSALP) gene was associated with the bone mineral density (BMD). BMD was the lowest among TNSALP 787T homozygotes (TT-type) and highest among TNSALP 787T>C homozygotes (CC-type) in postmenopausal women. In the present study, we investigated the effects of the TNSALP genotype on associations among serum bonespecific alkaline phosphatase (BAP), serum calcium, and phosphorus in healthy young Japanese subjects. Young healthy adult subjects (n=193) were genotyped for the polymorphism, and we measured the levels of serum BAP, serum calcium, and phosphorus. Dietary nutrient intakes were calculated based on 3-day food records before the day of blood examinations. Grouped by the TNSALP genotype, a significant negative correlation between serum BAP and phosphorus was observed in 787T>C homozygotes (CC-type), but not in heterozygotes (TCtype), nor in 787T homozygotes (TT-type). In the present study, we revealed that the single nucleotide polymorphism 787T>C in the TNSALP gene had effects on the correlation between serum BAP and phosphorus in young adult subjects. These results suggest that variation in TNSALP may be an important determinant of phosphate metabolism. Our data may be useful for planning strategies to prevent osteoporosis.

  17. [Alkaline phosphatase activity in blood group B or O secretors is fluctuated by the dinner intake of previous night].

    PubMed

    Matsushita, Makoto; Harajiri, Sanae; Tabata, Shiori; Yukimasa, Nobuyasu; Muramoto, Yoshimi; Komoda, Tsugikazu

    2013-04-01

    We previously reported that two intestinal alkaline phosphatase (IAP) isoforms, high molecular mass IAP (HIAP) and normal molecular mass IAP (NIAP), appear in healthy serum with our Triton-PAGE method for determination of ALP isozymes. In addition, HIAP is chiefly present in blood group B or O secretors, and a large amount of NIAP is secreted into the circulation after high-fat meal in blood group B or O secretors. In the present paper, we investigated the relationship between alkaline phosphatase (ALP) activity in early morning with the patient in a fasted state and the dinner intake of previous night. Two types of dinner were prepared; a low-fat meal (520 kcal), and a high-fat meal (1,040 kcal). Subjects ate the 2 types of dinner on different days. The mean ALP activities at 14 h after high-fat meal ingestion in blood group B or O secretors (n=14) from JSCC and IFCC methods were 8.8% and 5.2% higher than those at 14 h after low-fat meal ingestion in blood group B or O secretors, respectively. The increases in ALP activity between after high-fat meal and low-fat meal were nearly identical to the increases in NIAP activity. These results suggest that a high-fat meal is more likely to affect ALP activity at the early morning with the patient in a fasted state in blood group B or O secretors.

  18. Probing the origins of catalytic discrimination between phosphate and sulfate monoester hydrolysis: comparative analysis of alkaline phosphatase and protein tyrosine phosphatases.

    PubMed

    Andrews, Logan D; Zalatan, Jesse G; Herschlag, Daniel

    2014-11-04

    Catalytic promiscuity, the ability of enzymes to catalyze multiple reactions, provides an opportunity to gain a deeper understanding of the origins of catalysis and substrate specificity. Alkaline phosphatase (AP) catalyzes both phosphate and sulfate monoester hydrolysis reactions with a ∼10(10)-fold preference for phosphate monoester hydrolysis, despite the similarity between these reactions. The preponderance of formal positive charge in the AP active site, particularly from three divalent metal ions, was proposed to be responsible for this preference by providing stronger electrostatic interactions with the more negatively charged phosphoryl group versus the sulfuryl group. To test whether positively charged metal ions are required to achieve a high preference for the phosphate monoester hydrolysis reaction, the catalytic preference of three protein tyrosine phosphatases (PTPs), which do not contain metal ions, were measured. Their preferences ranged from 5 × 10(6) to 7 × 10(7), lower than that for AP but still substantial, indicating that metal ions and a high preponderance of formal positive charge within the active site are not required to achieve a strong catalytic preference for phosphate monoester over sulfate monoester hydrolysis. The observed ionic strength dependences of kcat/KM values for phosphate and sulfate monoester hydrolysis are steeper for the more highly charged phosphate ester with both AP and the PTP Stp1, following the dependence expected based on the charge difference of these two substrates. However, the dependences for AP were not greater than those of Stp1 and were rather shallow for both enzymes. These results suggest that overall electrostatics from formal positive charge within the active site is not the major driving force in distinguishing between these reactions and that substantial discrimination can be attained without metal ions. Thus, local properties of the active site, presumably including multiple positioned dipolar

  19. Characterization of soluble and membrane-bound alkaline phosphatase in Nilaparvata lugens and their potential relation to development and insecticide resistance.

    PubMed

    Wang, Zengxia; Liu, Shuhua; Yang, Baojun; Liu, Zewen

    2011-09-01

    Two forms (soluble and membrane-bound) of alkaline phosphatases (ALPs) were found in the brown planthopper, Nilaparvata lugens. In order to further study ALPs in N. lugens, two putative ALP genes (Nl-ALP1 and Nl-ALP2) were identified in this pest. Both Nl-ALP1 and Nl-ALP2 show approximately the same degree of sequence identity (40-50%) to other insect soluble and membrane-bound forms of ALP. Correlation of ALP activity and mRNA levels at different developmental stages, or following application of 20-hydroxyecdysone (20E) and insecticide fenvalerate, suggests that Nl-ALP1 and Nl-ALP2 might encode a soluble (sALP) and a membrane-bound ALP (mALP), respectively. Nl-ALP1-specific antibody Nl1-I detected only a specific band in soluble protein preparations and Nl-ALP2 specific antibody Nl2-I only detected a specific band in insoluble protein preparations, which provided conclusive linkages between Nl-ALP1 and a sALP and between Nl-ALP2 and a m ALP. Then, Nl-ALP1 was denoted as Nl-sALP for a sALP and Nl-ALP2 was denoted as Nl-mALP for a mALP. Only sALP activity and Nl-sALP mRNA level were induced by 20E and fenvalerate, which was confirmed by the density of specific band detected by Nl1-I in Sus strain with or without fenvalerate treatment. Additionally, the sALP activity, as well as Nl-sALP mRNA level, was significantly higher in a fenvalerate resistant population, compared with Sus strain. These results indicate that the sALP is more responsive to chemical stimulus, such as hormone and insecticide, and might play dual roles in development and insecticide tolerance. © 2011 Wiley Periodicals, Inc.

  20. ALP gene expression in cDNA samples from bone tissue engineering using a HA/TCP/Chitosan scaffold

    NASA Astrophysics Data System (ADS)

    Stephanie, N.; Katarina, H.; Amir, L. R.; Gunawan, H. A.

    2017-08-01

    This study examined the potential use of hydroxyapatite (HA)/tricalcium phosphate (TCP)/Chitosan as a bone tissue engineering scaffold. The potential for using HA/TCP/chitosan as a scaffold was analyzed by measuring expression of the ALP osteogenic gene in cDNA from bone biopsies from four Macaque nemestrina. Experimental conditions included control (untreated), treatment with HA/TCP 70:30, HA/TCP 50:50, and HA/TCP/chitosan. cDNA samples were measured quantitively with Real-Time PCR (qPCR) and semi-quantitively by gel electrophoresis. There were no significant differences in ALP gene expression between treatment subjects after two weeks, but the HA/TCP/chitosan treatment gave the highest level of expression after four weeks. The scaffold using the HA/TCP/chitosan combination induced a higher level of expression of the osteogenic gene ALP than did scaffold without chitosan.

  1. Use of bone alkaline phosphatase to monitor alendronate therapy in individual postmenopausal osteoporotic women.

    PubMed

    Kress, B C; Mizrahi, I A; Armour, K W; Marcus, R; Emkey, R D; Santora, A C

    1999-07-01

    Biochemical bone markers are sensitive to the changes in bone turnover that result from treatment of postmenopausal osteoporotic women with antiresorptive therapies. Although information is available on the use of bone markers in monitoring therapy in groups of subjects, less is known regarding how these markers perform in individual patients. Serum bone alkaline phosphatase (bone ALP) concentrations, measured with the Tandem(R) Ostase(R) assay, were used to monitor the biochemical response of bone in postmenopausal women with osteoporosis receiving either 10 mg/day alendronate therapy (n = 74) or calcium supplementation (n = 148) for 24 months. Bone ALP decreased significantly from baseline at 3 months (P ALP decrease in the treated osteoporotic population was consistent with normalization to premenopausal concentrations. Of the 74 alendronate-treated subjects, 63 (85.1%) demonstrated a decrease from baseline in bone ALP by 6 months that exceeded the least significant change of 25%. The bone ALP decrease from baseline exceeded 25% in 72 (97%) by the end of the study. The bone ALP assay is a sensitive and reliable tool that may be used to monitor the reduction in bone turnover after alendronate therapy in individual postmenopausal osteoporotic women.

  2. Dephosphorylation of microtubule-binding sites at the neurofilament-H tail domain by alkaline, acid, and protein phosphatases.

    PubMed

    Hisanaga, S; Yasugawa, S; Yamakawa, T; Miyamoto, E; Ikebe, M; Uchiyama, M; Kishimoto, T

    1993-06-01

    The dephosphorylation-induced interaction of neurofilaments (NFs) with microtubules (MTs) was investigated by using several phosphatases. Escherichia coli alkaline and wheat germ acid phosphatases increased the electrophoretic mobility of NF-H and NF-M by dephosphorylation, and induced the binding of NF-H to MTs. The binding of NFs to MTs was observed only after the electrophoretic mobility of NF-H approached the exhaustively dephosphorylated level when alkaline phosphatase was used. The number of phosphate remaining when NF-H began to bind to MTs was estimated by measuring phosphate bound to NF-H. NF-H did not bind to MTs even when about 40 phosphates from the total of 51 had been removed by alkaline phosphatase. The removal of 6 further phosphates finally resulted in the association of NF-H with MTs. A similar finding, that the restricted phosphorylation sites in the NF-H tail domain, but not the total amount of phosphates, were important for binding to MTs, was also obtained with acid phosphatases. In contrast to alkaline and acid phosphatases, four classes of protein phosphatases (protein phosphatases 1, 2A, 2B, and 2C) were ineffective for shifting the electrophoretic mobility of NF proteins and for inducing the association of NFs to MTs.

  3. Infantile hypophosphatasia combined with vitamin B6-responsive seizures and reticular formation lesions on magnetic resonance imaging: A case report.

    PubMed

    Fukazawa, Mitsuharu; Tezuka, Junichiro; Sasazuki, Momoko; Masumoto, Natsuko; Baba, Haruhisa; Doi, Takehiko; Tsutsumi, Yasushi; Mizuno, Yuji; Mihara, Futoshi; Nakayama, Hideki

    2018-02-01

    Hypophosphatasia (HPP) is a rare genetic disorder characterized by rachitic bone manifestations and a low serum alkaline phosphatase (ALP) level. It is caused by mutations in the tissue non-specific alkaline phosphatase (TNSALP) gene, which encodes the tissue non-specific isozyme of ALP. HPP patients exhibit various presentations depending on their age at onset, such as infantile HPP combined with vitamin B6-responsive seizures. A newborn with infantile HPP presented with tonic convulsions from day 5 after birth and received intravenous vitamin B6 (10mg/kg/day pyridoxal phosphate). Eleven days later, frequent apneic episodes occurred, and head magnetic resonance imaging (MRI) showed bilateral reticular formation lesions in the brain stem, including the medulla oblongata. After the pyridoxal phosphate dose was increased (to 40mg/kg/day), the patient's seizures and apnea resolved, and her MRI findings also improved. Genetic testing revealed that she was homozygous for the 1559delT mutation of TNSALP. High-dose pyridoxal phosphate is a useful treatment for HPP-induced seizures and might improve reticular formation lesions. Copyright © 2017 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.

  4. Alkaline and alkaline earth metal phosphate halides and phosphors

    DOEpatents

    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.

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

    PubMed

    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 <1500 g birth weight. Two blood samples, from each infant on at least 2 consecutive weeks, were reported for calcium, phosphorus, and ALP. Evidence of osteopenia was evaluated radiologically by performing wrist/knee x-ray.Sixteen infants (13.3%) had evidence of osteopenia in x-ray, whereas 104 infants (86.7%) were nonosteopenic and all the osteopenic infants were <1000-g birth weight. Birth weight and gestational age were significantly inversely related to serum ALP levels. Both samples showed statistically significantly higher mean ALP level in osteopenic than nonosteopenics (P < 0.001, and P < 0.001 respectively). There was no constant value of serum ALP related to radiologic evidence of osteopenia. However, the optimal cutoff value of serum ALP at which osteopenia is detected is 500 IU/L with 100% sensitivity and 80.77% specificity.High levels of ALP can be considered a reliable biomarker to predict the status of bone mineralization and the need for radiological evaluation in premature infants particularly those <1000-g birth weight and <32 weeks' gestation.

  6. Serum Alkaline Phosphatase Levels Predict Infection-Related Mortality and Hospitalization in Peritoneal Dialysis Patients.

    PubMed

    Hwang, Seun Deuk; Kim, Su-Hyun; Kim, Young Ok; Jin, Dong Chan; Song, Ho Chul; Choi, Euy Jin; Kim, Yong-Lim; Kim, Yon-Su; Kang, Shin-Wook; Kim, Nam-Ho; Yang, Chul Woo; Kim, Yong Kyun

    2016-01-01

    Serum alkaline phosphatase (ALP) levels have been reported to be associated with all-cause and cardiovascular mortality in peritoneal dialysis (PD) patients. However, it is unclear whether serum ALP levels predict infection-related clinical outcomes in PD patients. The aim of this study was to determine the relationships between serum ALP levels, infection-related mortality and hospitalization in PD patients. PD patients from the Clinical Research Center registry for end-stage renal disease, a multicenter prospective observational cohort study in Korea, were included in the present study. Patients were categorized into three groups by serum ALP tertiles as follows: Tertile 1, ALP <78 U/L; Tertile 2, ALP = 78-155 U/L; Tertile 3, ALP >155 U/L. Tertile 1 was used as the reference category. The primary outcomes were infection-related mortality and hospitalization. A total of 1,455 PD patients were included. The median follow-up period was 32 months. The most common cause of infection-related mortality and hospitalization was PD-related peritonitis. Multivariate Cox regression analyses showed that patients in the highest tertiles of serum ALP levels were at higher risk of infection-related mortality (HR 2.29, 95% CI, 1.42-5.21, P = 0.008) after adjustment for clinical variables. Higher tertiles of serum ALP levels were associated with higher risk of infection-related hospitalization (Tertile 2: HR 1.56, 95% CI, 1.18-2.19, P = 0.009, tertile 3: HR 1.34, 95% CI, 1.03-2.62, P = 0.031). Our data showed that elevated serum ALP levels were independently associated with a higher risk of infection-related mortality and hospitalization in PD patients.

  7. Osteosarcoma tissues and cell lines from patients with differing serum alkaline phosphatase concentrations display minimal differences in gene expression patterns

    PubMed Central

    de Sá Rodrigues, L. C.; Holmes, K. E.; Thompson, V.; Piskun, C. M.; Lana, S. E.; Newton, M. A.; Stein, T. J.

    2016-01-01

    Serum alkaline phosphatase (ALP) concentration is a prognostic factor for osteosarcoma in multiple studies, although its biological significance remains incompletely understood. To determine whether gene expression patterns differed in osteosarcoma from patients with differing serum ALP concentrations, microarray analysis was performed on 18 primary osteosarcoma samples and six osteosarcoma cell lines from dogs with normal and increased serum ALP concentration. No differences in gene expression patterns were noted between tumours or cell lines with differing serum ALP concentration using a gene-specific two-sample t-test. Using a more sensitive empirical Bayes procedure, defective in cullin neddylation 1 domain containing 1 (DCUN1D1) was increased in both the tissue and cell lines of the normal ALP group. Using quantitative PCR (qPCR), differences in DCUN1D1 expression between the two groups failed to reach significance. The homogeneity of gene expression patterns of osteosarcoma associated differing serum ALP concentrations are consistent with previous studies suggesting serum ALP concentration is not associated with intrinsic differences of osteosarcoma cells. PMID:25643733

  8. Induction of rat alkaline phosphatase isozymes bearing a glycan-phosphatidylinositol anchor modified by in vivo treatment with a benzimidazole derivative linked to ethylbenzene.

    PubMed

    Harada, T; Koyama, I; Sato, K; Komoda, T

    2000-10-01

    Serum alkaline phosphatase (ALP) is detected in soluble-form as a result of translocation from the membrane site by cleavage at the glycosyl-phosphatidylinositol moiety (GPI anchor). It is known that membrane-bound ALP (mALP) can be detected in serum in certain pathological and physiological conditions, and that it can be solubilized in vitro to soluble-ALP (sALP) by phosphatidylinositol-specific phospholipase C (PIPLC), phospholipase D, bile salt, detergent, etc. We observed a marked increase in ALP activity in the serum of rats given a benzimidazole derivative by gavage, and detected it as slow-migrating ALPs (SM-ALPs), which were mALP-like but resistant to PIPLC and n-butanol treatment on disc PAGE. On the other hand, ficin treatment made SM-ALPs shift to the sALP position. The molecular size of the SM-ALPs was smaller than that of sALP on sodium dodecyl sulphide-polyacrylamide slab-gel electrophoresis (SDS-PAGE), and immunoreactivity revealed the intestinal type. SM-ALPs were also detected in the duodenum and jejunum. The main sugar chain structure of SM-ALPs was the biantennary complex-type, which was coincided with intestinal sALP sugar chain. These results suggest that intestinal ALPs induced by the benzimidazole derivative were modified in their C-terminus or GPI anchor region and modification of this region may also participate in translocation into the bloodstream.

  9. Gingival crevicular fluid protein content and alkaline phosphatase activity in relation to pubertal growth phase.

    PubMed

    Perinetti, Giuseppe; Franchi, Lorenzo; Castaldo, Attilio; Contardo, Luca

    2012-11-01

    To evaluate gingival crevicular fluid (GCF) protein content and alkaline phosphatase (ALP) activity in growing subjects in relation to stages of skeletal maturation, ie, the growth phase, as prepubertal, pubertal, and postpubertal. Fifty healthy growing subjects (31 girls and 19 boys; age range, 7.8-17.7 years) were enrolled in this study that followed a double-blind, prospective, cross-sectional design. Collection of GCF was performed at the mesial and distal sites of both central incisors, for the maxilla and mandible. Growth phase was assessed through the cervical vertebral maturation method. GCF parameters were expressed as total protein content, total ALP activity, and normalized ALP activity. The total GCF protein content was similar between the different growth phases. On the contrary, the total ALP activity showed a peak for the pubertal growth phase. The normalized GCF ALP activity was only poorly associated with growth phase. No differences were seen between the maxillary and mandibular sites, or between the sexes, for any GCF parameter. The total GCF protein content is not sensitive to the growth phase. However, GCF ALP activity has potential as a diagnostic aid for identification of the pubertal growth phase in individual subjects when expressed as total, but not normalized, values.

  10. Extracellular Ca2(+)-dependent inducible alkaline phosphatase from extremely halophilic archaebacterium Haloarcula marismortui.

    PubMed Central

    Goldman, S; Hecht, K; Eisenberg, H; Mevarech, M

    1990-01-01

    When starved of inorganic phosphate, the extremely halophilic archaebacterium Haloarcula marismortui produces the enzyme alkaline phosphatase and secretes it to the medium. This inducible extracellular enzyme is a glycoprotein whose subunit molecular mass is 160 kDa, as estimated by sodium dodecyl sulfate-gel electrophoresis. The native form of the enzyme is heterogeneous and composed of multiple oligomeric forms. The enzymatic activity of the halophilic alkaline phosphatase is maximal at pH 8.5, and the enzyme is inhibited by phosphate. Unlike most alkaline phosphatases, the halobacterial enzyme requires Ca2+ and not Zn2+ ions for its activity. Both calcium ions (in the millimolar range) and NaCl (in the molar range) are required for the stability of the enzyme. Images PMID:2123861

  11. Effects of Pore Size on the Osteoconductivity and Mechanical Properties of Calcium Phosphate Cement in a Rabbit Model.

    PubMed

    Zhao, Yi-Nan; Fan, Jun-Jun; Li, Zhi-Quan; Liu, Yan-Wu; Wu, Yao-Ping; Liu, Jian

    2017-02-01

    Calcium phosphate cement (CPC) porous scaffold is widely used as a suitable bone substitute to repair bone defect, but the optimal pore size is unclear yet. The current study aimed to evaluate the effect of different pore sizes on the processing of bone formation in repairing segmental bone defect of rabbits using CPC porous scaffolds. Three kinds of CPC porous scaffolds with 5 mm diameters and 12 mm length were prepared with the same porosity but different pore sizes (Group A: 200-300 µm, Group B: 300-450 µm, Group C: 450-600 µm, respectively). Twelve millimeter segmental bone defects were created in the middle of the radius bone and filled with different kinds of CPC cylindrical scaffolds. After 4, 12, and 24 weeks, alkaline phosphatase (ALP), histological assessment, and mechanical properties evaluation were performed in all three groups. After 4 weeks, ALP activity increased in all groups but was highest in Group A with smallest pore size. The new bone formation within the scaffolds was not obvious in all groups. After 12 weeks, the new bone formation within the scaffolds was obvious in each group and highest in Group A. At 24 weeks, no significant difference in new bone formation was observed among different groups. Besides the osteoconductive effect, Group A with smallest pore size also had the best mechanical properties in vivo at 12 weeks. We demonstrate that pore size has a significant effect on the osteoconductivity and mechanical properties of calcium phosphate cement porous scaffold in vivo. Small pore size favors the bone formation in the early stage and may be more suitable for repairing segmental bone defect in vivo. © 2016 International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  12. Could Alerting Physicians for Low Alkaline Phosphatase Levels Be Helpful in Early Diagnosis of Hypophosphatasia?

    PubMed

    Deeb, Asma; Elfatih, Abubaker

    2018-03-01

    Hypophosphatasia (HPP) is an inborn error of metabolism with significant morbidity and mortality. Its presentation is nonspecific leading to delayed or missed diagnosis. Low alkaline phosphatase (ALP) is a diagnostic test. Unlike high ALP, low level is commonly not flagged by laboratories as abnormal. A new treatment was shown to be effective in HPP. In this study we aimed to establish the frequency of low ALP levels requiring notification to physicians by the laboratory and also to describe the clinical manifestations of patients presenting with low ALP for a possible diagnosis of HPP. Patients under age 18 years with low ALP levels were identified from biochemistry records over a period of 6 months. Reference ranges were used as per the Associated Regional and University Pathologists Reference Laboratory (Utah, USA). Electronic results for patients with low levels were checked for flagging as abnormal/low ALP results. Charts of identified patients were reviewed. Presenting features were categorized under groups of disorders. ALP levels were tested in 2890 patients. 702 had values less than 160 U/L. Of these patients, 226 (32%) had age/gender specific low ALP. None of the low ALP results was flagged as low. Twenty-one had more than one low reading and their charts were reviewed. Four patients in the neuromuscular and four in the miscellaneous group presented with features consistent with HPP despite these patients having no specific diagnoses. Laboratories do not alert physicians in cases with low ALP levels. A persistently low level in patients with unspecified diagnoses could be a key to diagnose HPP. Implementing lab-specific ranges and alerting for low levels could prompt physicians to investigate for undiagnosed HPP.

  13. Interdependency of fission yeast Alp14/TOG and coiled coil protein Alp7 in microtubule localization and bipolar spindle formation.

    PubMed

    Sato, Masamitsu; Vardy, Leah; Angel Garcia, Miguel; Koonrugsa, Nirada; Toda, Takashi

    2004-04-01

    The Dis1/TOG family plays a pivotal role in microtubule organization. In fission yeast, Alp14 and Dis1 share an essential function in bipolar spindle formation. Here, we characterize Alp7, a novel coiled-coil protein that is required for organization of bipolar spindles. Both Alp7 and Alp14 colocalize to the spindle pole body (SPB) and mitotic spindles. Alp14 localization to these sites is fully dependent upon Alp7. Conversely, in the absence of Alp14, Alp7 localizes to the SPBs, but not mitotic spindles. Alp7 forms a complex with Alp14, where the C-terminal region of Alp14 interacts with the coiled-coil domain of Alp7. Intriguingly, this Alp14 C terminus is necessary and sufficient for mitotic spindle localization. Overproduction of either full-length or coiled-coil region of Alp7 results in abnormal V-shaped spindles and stabilization of interphase microtubules, which is induced independent of Alp14. Alp7 may be a functional homologue of animal TACC. Our results shed light on an interdependent relationship between Alp14/TOG and Alp7. We propose a two-step model that accounts for the recruitment of Alp7 and Alp14 to the SPB and microtubules.

  14. Tumour-derived alkaline phosphatase regulates tumour growth, epithelial plasticity and disease-free survival in metastatic prostate cancer

    PubMed Central

    Rao, S R; Snaith, A E; Marino, D; Cheng, X; Lwin, S T; Orriss, I R; Hamdy, F C; Edwards, C M

    2017-01-01

    Background: Recent evidence suggests that bone-related parameters are the main prognostic factors for overall survival in advanced prostate cancer (PCa), with elevated circulating levels of alkaline phosphatase (ALP) thought to reflect the dysregulated bone formation accompanying distant metastases. We have identified that PCa cells express ALPL, the gene that encodes for tissue nonspecific ALP, and hypothesised that tumour-derived ALPL may contribute to disease progression. Methods: Functional effects of ALPL inhibition were investigated in metastatic PCa cell lines. ALPL gene expression was analysed from published PCa data sets, and correlated with disease-free survival and metastasis. Results: ALPL expression was increased in PCa cells from metastatic sites. A reduction in tumour-derived ALPL expression or ALP activity increased cell death, mesenchymal-to-epithelial transition and reduced migration. Alkaline phosphatase activity was decreased by the EMT repressor Snail. In men with PCa, tumour-derived ALPL correlated with EMT markers, and high ALPL expression was associated with a significant reduction in disease-free survival. Conclusions: Our studies reveal the function of tumour-derived ALPL in regulating cell death and epithelial plasticity, and demonstrate a strong association between ALPL expression in PCa cells and metastasis or disease-free survival, thus identifying tumour-derived ALPL as a major contributor to the pathogenesis of PCa progression. PMID:28006818

  15. Phosphate sorption and desorption on pyrite in primitive aqueous scenarios: relevance of acidic --> alkaline transitions.

    PubMed

    de Souza-Barros, Fernando; Braz-Levigard, Raphael; Ching-San, Yonder; Monte, Marisa M B; Bonapace, José A P; Montezano, Viviane; Vieyra, Adalberto

    2007-02-01

    Phosphate (P(i)) sorption assays onto pyrite in media simulating primeval aquatic scenarios affected by hydrothermal emissions, reveal that acidic conditions favour P(i) sorption whereas mild alkaline media--as well as those simulating sulfur oxidation to SO(2-) (4)--revert this capture process. Several mechanisms relevant to P(i) availability in prebiotic eras are implicated in the modulation of these processes. Those favouring sorption are: (a) hydrophobic coating of molecules, such as acetate that could be formed in the vicinity of hydrothermal vents; (b) water and Mg(2+) bridging in the interface mineral-aqueous media; (c) surface charge neutralization by monovalent cations (Na+ and K+). The increase of both the medium pH and the SO(2-) (4) trapping by the mineral interface would provoke the release of sorbed P(i) due to charge polarization. Moreover it is shown that P(i) self-modulates its sorption, a mechanism that depends on the abundance of SO(2-) (4) in the interface. The relevance of the proposed mechanisms of P(i) capture, release and trapping arises from the need of abundant presence of this molecule for primitive phosphorylations, since--similarly to contemporary aqueous media--inorganic phosphate concentrations in primitive seas should have been low. It is proposed that the presence of sulphide minerals with high affinity to P(i) could have trapped this molecule in an efficient manner, allowing its concentration in specific niches. In these niches, the conditions studied in the present work would have been relevant for its availability in soluble form, specially in primitive insulated systems with pH gradients across the wall.

  16. Is alkaline phosphatase the smoking gun for highly refractory primitive leukemic cells?

    PubMed Central

    Rico, Laura G.; Juncà, Jordi; Ward, Mike D.; Bradford, Jolene; Petriz, Jordi

    2016-01-01

    With the aim to detect candidate malignant primitive progenitor populations, we modified an original alkaline phosphatase (ALP) stem cell detection method based on the identification of alkaline phosphatase fluorescent cells in combination with flow cytometry immunophenotyping. Over a period of one year, we have been using this technique to study its activity in patients with leukemia and lymphoma, showing that changes in the alkaline phosphatase levels can be used to detect rare populations of highly refractory malignant cells. By screening different blood cancers, we have observed that this activity is not always restricted to CD34+ leukemic cells, and can be overexpressed in CD34 negative leukemia. We have verified that this method gives accurate and reproducible measurements and our preliminary results suggest that CD34+/ALPhigh cells appear to sustain leukemogenesis over time. PMID:27732563

  17. Interdependency of Fission Yeast Alp14/TOG and Coiled Coil Protein Alp7 in Microtubule Localization and Bipolar Spindle FormationD⃞

    PubMed Central

    Sato, Masamitsu; Vardy, Leah; Angel Garcia, Miguel; Koonrugsa, Nirada; Toda, Takashi

    2004-01-01

    The Dis1/TOG family plays a pivotal role in microtubule organization. In fission yeast, Alp14 and Dis1 share an essential function in bipolar spindle formation. Here, we characterize Alp7, a novel coiled-coil protein that is required for organization of bipolar spindles. Both Alp7 and Alp14 colocalize to the spindle pole body (SPB) and mitotic spindles. Alp14 localization to these sites is fully dependent upon Alp7. Conversely, in the absence of Alp14, Alp7 localizes to the SPBs, but not mitotic spindles. Alp7 forms a complex with Alp14, where the C-terminal region of Alp14 interacts with the coiled-coil domain of Alp7. Intriguingly, this Alp14 C terminus is necessary and sufficient for mitotic spindle localization. Overproduction of either full-length or coiled-coil region of Alp7 results in abnormal V-shaped spindles and stabilization of interphase microtubules, which is induced independent of Alp14. Alp7 may be a functional homologue of animal TACC. Our results shed light on an interdependent relationship between Alp14/TOG and Alp7. We propose a two-step model that accounts for the recruitment of Alp7 and Alp14 to the SPB and microtubules. PMID:14742702

  18. ALPS - A LINEAR PROGRAM SOLVER

    NASA Technical Reports Server (NTRS)

    Viterna, L. A.

    1994-01-01

    Linear programming is a widely-used engineering and management tool. Scheduling, resource allocation, and production planning are all well-known applications of linear programs (LP's). Most LP's are too large to be solved by hand, so over the decades many computer codes for solving LP's have been developed. ALPS, A Linear Program Solver, is a full-featured LP analysis program. ALPS can solve plain linear programs as well as more complicated mixed integer and pure integer programs. ALPS also contains an efficient solution technique for pure binary (0-1 integer) programs. One of the many weaknesses of LP solvers is the lack of interaction with the user. ALPS is a menu-driven program with no special commands or keywords to learn. In addition, ALPS contains a full-screen editor to enter and maintain the LP formulation. These formulations can be written to and read from plain ASCII files for portability. For those less experienced in LP formulation, ALPS contains a problem "parser" which checks the formulation for errors. ALPS creates fully formatted, readable reports that can be sent to a printer or output file. ALPS is written entirely in IBM's APL2/PC product, Version 1.01. The APL2 workspace containing all the ALPS code can be run on any APL2/PC system (AT or 386). On a 32-bit system, this configuration can take advantage of all extended memory. The user can also examine and modify the ALPS code. The APL2 workspace has also been "packed" to be run on any DOS system (without APL2) as a stand-alone "EXE" file, but has limited memory capacity on a 640K system. A numeric coprocessor (80X87) is optional but recommended. The standard distribution medium for ALPS is a 5.25 inch 360K MS-DOS format diskette. IBM, IBM PC and IBM APL2 are registered trademarks of International Business Machines Corporation. MS-DOS is a registered trademark of Microsoft Corporation.

  19. International symposium on cellular and molecular biology of phosphate and phosphorylated compounds in microorganisms: Proceedings

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    NONE

    1993-12-31

    This report contains the abstracts of papers presented at the conference. Attention is focused on the following topics: regulation of phosphate metabolism in bacteria; structure-function of alkaline phosphatase; regulation of phosphate metabolism in yeast; transport of phosphate and phosphorylated compounds; and phosphate regulation in pathogenesis and secondary metabolism.

  20. Baseline Serum Clinical Chemistry Values in African Green Monkeys Before and After Sulfur Mustard

    DTIC Science & Technology

    2007-05-01

    aspartate transaminase (189 %), blood urea nitrogen (75 %), creatine kinase (721 %), and lactate dehydrogenase (114 %) one day after HD exposure...ALT, 93 %), aspartate transaminase (AST, 189 %), blood urea nitrogen (BUN, 75 %), creatine kinase (CK, 721 %), and lactate dehydrogenase (LDH, 114...alkaline phosphate (ALP), alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin (TBIL), calcium (Ca2+), creatine kinase (CK

  1. Effect of alkaline earth modifier on the optical and structural properties of Cu2+ doped phosphate glasses as a bandpass filter

    NASA Astrophysics Data System (ADS)

    Farouk, M.; Samir, A.; El Okr, M.

    2018-02-01

    Glasses of composition [16RO-3Al2O3sbnd 6CuOsbnd 20Na2Osbnd 55P2O5], where R is the alkaline earth (R = Mg, Ca, Sr and Ba mol. %), were prepared by conventional melt quenching technique. The glass samples were characterized by X-ray diffraction, infrared spectroscopy, and spectrophotometer. XRD patterns show no sharp peaks indicating the non-crystalline nature of the prepared glasses. The density and molar volume of the glass systems were determined in order to study their structures. These results revealed that addition of alkaline earth elements leads to the formation of non-bridging oxygens (NBOs) and expands (opens up) the structure. The infrared spectra were analyzed to quantify the present phosphate groups. The optical absorption spectra of Cu2+ ions show the characteristic broadband single of Cu2+ ions in octahedral symmetry. The band gap was estimated following two methodologies. The first method considers the band edge of the transmission, while the second approach relays on the estimated values of the optical constants. A decent agreement for the band gap values using the two methods was obtained.

  2. Catalytic efficiency is a better predictor of arsenic toxicity to soil alkaline phosphatase.

    PubMed

    Wang, Ziquan; Tian, Haixia; Lu, Guannan; Zhao, Yiming; Yang, Rui; Megharaj, Mallavarapu; He, Wenxiang

    2018-02-01

    Arsenic (As) is an inhibitor of phosphatase, however, in the complex soil system, the substrate concentration effect and the mechanism of As inhibition of soil alkaline phosphatase (ALP) and its kinetics has not been adequately studied. In this work, we investigated soil ALP activity in response to As pollution at different substrate concentrations in various types of soils and explored the inhibition mechanism using the enzyme kinetics. The results showed that As inhibition of soil ALP activity was substrate concentration-dependent. Increasing substrate concentration decreased inhibition rate, suggesting reduced toxicity. This dependency was due to the competitive inhibition mechanism of As to soil ALP. The kinetic parameters, maximum reaction velocity (V max ) and Michaelis constant (K m ) in unpolluted soils were 0.012-0.267mMh -1 and 1.34-3.79mM respectively. The competitive inhibition constant (K ic ) was 0.17-0.70mM, which was lower than K m , suggesting higher enzyme affinity for As than for substrate. The ecological doses, ED 10 and ED 50 (concentration of As that results in 10% and 50% inhibition on enzyme parameter) for inhibition of catalytic efficiency (V max /K m ) were lower than those for inhibition of enzyme activity at different substrate concentrations. This suggests that the integrated kinetic parameter, catalytic efficiency is substrate concentration independent and more sensitive to As than ALP activity. Thus, catalytic efficiency was proposed as a more reliable indicator than ALP activity for risk assessment of As pollution. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Biocontrol activity of an alkaline serine protease from Aureobasidium pullulans expressed in Pichia pastoris against four postharvest pathogens on apple.

    PubMed

    Banani, Houda; Spadaro, Davide; Zhang, Dianpeng; Matic, Slavica; Garibaldi, Angelo; Gullino, Maria Lodovica

    2014-07-16

    The yeast-like fungus Aureobasidium pullulans PL5 is a microbial antagonist against postharvest pathogens of fruits. The strain is able to produce hydrolases, including glucanases, chitinases and proteases. The alkaline serine protease gene ALP5 from A. pullulans was cloned, inserted into the vector pPIC9 to construct pPIC9/ALP5, and then expressed in Pichia pastoris strain KM71. ALP5 had a molecular mass of 42.9kDa after 5days growth with 1% methanol induction at 28°C. The recombinant protease expressed in P. pastoris showed its highest activity under alkaline conditions (at pH10) and a temperature of 50°C. The antifungal activity of the recombinant protease was investigated against Penicillium expansum, Botrytis cinerea, Monilinia fructicola and Alternaria alternata in vitro and on apple. The recombinant protease reduced significantly the spore germination and the germ tube length of the tested pathogens in PDB medium. The highest level of protease efficacy was observed against M. fructicola and B. cinerea, whereas a lower efficacy was observed against P. expansum and A. alternata indicating a possible effect of the pathogen cell wall composition on the proteolytic activity of the recombinant protease. The presence of protease was able to cause the swelling of the hyphae of B. cinerea, under an optical microscope. The recombinant protease expressed in P. pastoris was more active against the pathogens in vitro than the same enzyme expressed in E. coli in previous studies. The efficacy of ALP5 was also evaluated against the pathogens in vivo on cv Golden Delicious apples. The protease was more efficient in controlling M. fructicola, B. cinerea and P. expansum than A. alternata. However, the extent of the activity was dependent on the enzyme concentration and the length of fruit storage. This study demonstrated the capacity of the alkaline serine protease to keep its enzymatic activity for some days in the unfavorable environment of the fruit wounds. The alkaline

  4. Host sensitized near-infrared emission in Nd3+ doped different alkaline-sodium-phosphate phosphors

    NASA Astrophysics Data System (ADS)

    Balakrishna, A.; Swart, H. C.; Kroon, R. E.; Ntwaeaborwa, O. M.

    2018-04-01

    Near-infrared (NIR) emitting phosphors of different alkaline based sodium-phosphate (MNa[PO4], where M = Mg, Ca, Sr and Ba were prepared by a conventional solution combustion method with fixed doping concentration of Nd3+ (1.0 mol%). The phosphors were characterized by powder X-ray diffraction, field emission scanning electron microscope, Fourier transform infrared spectroscopy, UV-vis spectroscopy and fluorescent spectrophotometry. The optical properties including reflectance, excitation and emission were investigated. The excitation spectra of the phosphors were characterized by a broadband extending from 450 to 900 nm. Upon excitation with a wavelength of 580 nm, the phosphor emits intensely infrared region at 872 nm, 1060 nm and 1325 nm which correspond to the 4F3/2 → 4I9/2, 4F3/2 → 4I11/2 and 4F3/2 → 4I13/2 transitions of Nd3+ ions and were found to vary for the different hosts. The strongest emission wavelength reaches 1060 nm. The most intense emission of Nd3+ was observed from Ca2+ incorporated host. The down conversion emissions of the material fall in the NIR region suggesting that the prepared phosphors have potential application in the development of photonic devices emitting in the NIR.

  5. The route of non-enzymic and enzymic breakdown of 5-phosphoribosyl 1-pyrophosphate to ribose 1-phosphate.

    PubMed Central

    Trembacz, H; Jezewska, M M

    1990-01-01

    Spontaneous decomposition of 5-phosphoribosyl 1-pyrophosphate at pH 5.5 was established to occur as follows: 5-Phosphoribosyl 1-pyrophosphate----5-phosphoribosyl 1,2-(cyclic)phosphate----ribose 1-phosphate----ribose Enzymic degradation of 5-phosphoribosyl 1-pyrophosphate by alkaline phosphatase from calf intestine and by acid phosphatases from potato and Aspergillus niger was found to proceed according to this pathway within the pH range 2.5-7.4 with accumulation of ribose 1-phosphate. In the case of alkaline phosphatase, Mg2+ ions inhibit the pyrophosphorolysis of 5-phosphoribosyl 1-pyrophosphate and stimulate the hydrolysis of ribose 1-phosphate. PMID:1700897

  6. 1-alpha,25-Dihydroxyvitamin D3 up-regulates the expression of 2 types of human intestinal alkaline phosphatase alternative splicing variants in Caco-2 cells and may be an important regulator of their expression in gut homeostasis.

    PubMed

    Noda, Seiko; Yamada, Asako; Nakaoka, Kanae; Goseki-Sone, Masae

    2017-10-01

    Vitamin D insufficiency is associated with a greater risk of osteoporosis and also influences skeletal muscle functions, differentiation, and development. The principal function of vitamin D in calcium homeostasis is to increase the absorption of calcium from the intestine, and the level of alkaline phosphatase (ALP) activity, a differentiation marker for intestinal epithelial cells, is regulated by vitamin D. Intestinal-type ALP is expressed at a high concentration in the brush border membrane of intestinal epithelial cells, and is known to be affected by several kinds of nutrients. Recent reviews have highlighted the importance of intestinal-type ALP in gut homeostasis. Intestinal-type ALP controls bacterial endotoxin-induced inflammation by dephosphorylating lipopolysaccharide and is a gut mucosal defense factor. In this study, we investigated the influence of vitamin D on the expression of 2 types of alternative mRNA variants encoding the human alkaline phosphatase, intestinal (ALPI) gene in human Caco-2 cells as an in vitro model of the small intestinal epithelium. After treatment with 1-alpha,25-dihydroxyvitamin D 3 , the biologically active form of vitamin D 3 , there were significant increases in the ALP activities of Caco-2 cells. Inhibitor and thermal inactivation experiments showed that the increased ALP had properties of intestinal-type ALP. Reverse transcription-polymerase chain reaction analysis revealed that expression of the 2 types of alternative mRNA variants from the ALPI gene was markedly enhanced by vitamin D in Caco-2 cells. In conclusion, these findings agree with the hypothesis: vitamin D up-regulated the expression of 2 types of human intestinal alkaline phosphatase alternative splicing variants in Caco-2 cells; vitamin D may be an important regulator of ALPI gene expression in gut homeostasis. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Mineral induced formation of sugar phosphates

    NASA Technical Reports Server (NTRS)

    Pitsch, S.; Eschenmoser, A.; Gedulin, B.; Hui, S.; Arrhenius, G.

    1995-01-01

    Glycolaldehyde phosphate, sorbed from highly dilute, weakly alkaline solution into the interlayer of common expanding sheet structure metal hydroxide minerals, condenses extensively to racemic aldotetrose-2, 4-diphophates, and aldohexose-2, 4, 6-triphosphates. The reaction proceeds mainly through racemic erythrose-2, 4-phosphate, and terminates with a large fraction of racemic altrose-2, 4, 6-phosphate. In the absence of an inductive mineral phase, no detectable homogeneous reaction takes place in the concentration- and pH range used. The reactant glycolaldehyde phosphate is practically completely sorbed within an hour from solutions with concentrations as low as 50 micron; the half-time for conversion to hexose phosphates is of the order of two days at room temperature and pH 9.5. Total production of sugar phosphates in the mineral interlayer is largely independent of the glycolaldehyde phosphate concentration in the external solution, but is determined by the total amount of GAP offered for sorption up to the capacity of the mineral. In the presence of equimolar amounts of rac-glyceraldehyde-2-phosphate, but under otherwise similar conditions, aldopentose-2, 4, -diphosphates also form, but only as a small fraction of the hexose-2, 4, 6-phosphates.

  8. ALPS: A Linear Program Solver

    NASA Technical Reports Server (NTRS)

    Ferencz, Donald C.; Viterna, Larry A.

    1991-01-01

    ALPS is a computer program which can be used to solve general linear program (optimization) problems. ALPS was designed for those who have minimal linear programming (LP) knowledge and features a menu-driven scheme to guide the user through the process of creating and solving LP formulations. Once created, the problems can be edited and stored in standard DOS ASCII files to provide portability to various word processors or even other linear programming packages. Unlike many math-oriented LP solvers, ALPS contains an LP parser that reads through the LP formulation and reports several types of errors to the user. ALPS provides a large amount of solution data which is often useful in problem solving. In addition to pure linear programs, ALPS can solve for integer, mixed integer, and binary type problems. Pure linear programs are solved with the revised simplex method. Integer or mixed integer programs are solved initially with the revised simplex, and the completed using the branch-and-bound technique. Binary programs are solved with the method of implicit enumeration. This manual describes how to use ALPS to create, edit, and solve linear programming problems. Instructions for installing ALPS on a PC compatible computer are included in the appendices along with a general introduction to linear programming. A programmers guide is also included for assistance in modifying and maintaining the program.

  9. Differences of bone alkaline phosphatase isoforms in metastatic bone disease and discrepant effects of clodronate on different skeletal sites indicated by the location of pain.

    PubMed

    Magnusson, P; Larsson, L; Englund, G; Larsson, B; Strang, P; Selin-Sjögren, L

    1998-08-01

    We compared clodronate with placebo administration in 42 primarily or secondarily hormone-refractory prostate cancer patients with skeletal metastases and persisting pain. Serum total alkaline phosphatase (ALP), bone ALP isoforms, osteocalcin, cross-linked carboxy-terminal telopeptide of type I collagen, and prostate-specific antigen were analyzed before and after 1 month of treatment. Six ALP isoforms were quantified by HPLC: one bone/intestinal, two bone (B1, B2), and three liver ALP isoforms. The most apparent difference compared with healthy males was observed for the bone ALP isoform B2. Patients and healthy males had a B2 activity corresponding to 75% and 35% of the total ALP activity, respectively (P <0.0001). We propose that the different bone ALP isoforms reflect different stages of osteoblast differentiation during the extracellular matrix maturation phase of osteogenesis. All bone markers except osteocalcin increased after 1 month of clodronate administration. These increases were associated with pain only in the upper part of the body. We suggest that the uptake of clodronate by the skeleton was not uniform during our treatment period.

  10. Increased serum alkaline phosphatase levels correlate with high disease activity and low bone mineral density in patients with axial spondyloarthritis.

    PubMed

    Kang, Kwi Young; Hong, Yeon Sik; Park, Sung-Hwan; Ju, Ji Hyeon

    2015-10-01

    Recent studies report an association between serum alkaline phosphatase (ALP) levels and inflammation. The present study examined the relationship between ALP and disease activity, bone mineral density (BMD), and radiological damage in axial spondyloarthritis (SpA). A total of 115 patients who fulfilled the ASAS axial SpA criteria were enrolled. Serum ALP, bone-specific ALP (BALP), serum cross-linked telopeptide of type-I collagen (sCTX), and inflammatory markers were measured. Clinical parameters, BMD, grade of sacroiliitis, and the modified Stoke AS Spinal Score (mSASSS) were also assessed. The Ankylosing Spondylitis Disease Activity Score (ASDAS) was also calculated. The associations between serum ALP, disease activity score, BMD, and radiologic damage were evaluated. The mean serum ALP level was 77 ± 26 U/l. Serum ALP levels increased in 14 patients (13%). Serum ALP levels increased along with ASDAS-CRP after adjusting for age and sex (p = 0.004), and were significantly correlated with ASDAS-ESR and ASDAS-CRP (p = 0.001 and p < 0.001, respectively), negatively correlated with BMD in the lumbar spine and femoral neck (p = 0.003 and 0.046, respectively), and positively correlated with sacroiliitis grade and the mSASSS (p < 0.001 and p < 0.002, respectively). BALP and sCTX were not associated with disease activity or BMD. Multivariate analysis showed that serum ALP was independently associated with ASDAS-CRP (p = 0.010). Increased serum ALP levels were associated with high disease activity, low BMD, and higher structural damage scores in SpA patients. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Raloxifene microsphere-embedded collagen/chitosan/β-tricalcium phosphate scaffold for effective bone tissue engineering.

    PubMed

    Zhang, Ming-Lei; Cheng, Ji; Xiao, Ye-Chen; Yin, Ruo-Feng; Feng, Xu

    2017-02-25

    Engineering novel scaffolds that can mimic the functional extracellular matrix (ECM) would be a great achievement in bone tissue engineering. This paper reports the fabrication of novel collagen/chitosan/β-tricalcium phosphate (CCTP) based tissue engineering scaffold. In order to improve the regeneration ability of scaffold, we have embedded raloxifene (RLX)-loaded PLGA microsphere in the CCTP scaffold. The average pore of scaffold was in the range of 150-200μm with ideal mechanical strength and swelling/degradation characteristics. The release rate of RLX from the microsphere (MS) embedded scaffold was gradual and controlled. Also a significantly enhanced cell proliferation was observed in RLX-MS exposed cell group suggesting that microsphere/scaffold could be an ideal biomaterial for bone tissue engineering. Specifically, RLX-MS showed a significantly higher Alizarin red staining indicating the higher mineralization capacity of this group. Furthermore, a high alkaline phosphatase (ALP) activity for RLX-MS exposed group after 15days incubation indicates the bone regeneration capacity of MC3T3-E1 cells. Overall, present study showed that RLX-loaded microsphere embedded scaffold has the promising potential for bone tissue engineering applications. Copyright © 2016. Published by Elsevier B.V.

  12. Biochemical and Thermodynamical Characterization of Glucose Oxidase, Invertase, and Alkaline Phosphatase Secreted by Antarctic Yeasts.

    PubMed

    Yuivar, Yassef; Barahona, Salvador; Alcaíno, Jennifer; Cifuentes, Víctor; Baeza, Marcelo

    2017-01-01

    The use of enzymes in diverse industries has increased substantially over past decades, creating a well-established and growing global market. Currently, the use of enzymes that work better at ambient or lower temperatures in order to decrease the temperatures of production processes is desirable. There is thus a continuous search for enzymes in cold environments, especially from microbial sources, with amylases, proteases, lipases and, cellulases being the most studied. Other enzymes, such as glucose oxidase (GOD), invertase (Inv), and alkaline phosphatase (ALP), also have a high potential for application, but have been much less studied in microorganisms living in cold-environments. In this work, secretion of these three enzymes by Antarctic yeast species was analyzed, and five, three, and five species were found to produce extracellular GOD, Inv, and ALP, respectively. The major producers of GOD, Inv, and ALP were Goffeauzyma gastrica, Wickerhamomyces anomalus , and Dioszegia sp., respectively, from which the enzymes were purified and characterized. Contrary to what was expected, the highest GOD and Inv activities were found at 64°C and 60°C, respectively, and at 47°C for ALP. However, the three enzymes maintained a significant percentage of activity at lower temperatures, especially ALP that kept a 67 and 43% of activity at 10°C and 4°C, respectively.

  13. Biochemical and Thermodynamical Characterization of Glucose Oxidase, Invertase, and Alkaline Phosphatase Secreted by Antarctic Yeasts

    PubMed Central

    Yuivar, Yassef; Barahona, Salvador; Alcaíno, Jennifer; Cifuentes, Víctor; Baeza, Marcelo

    2017-01-01

    The use of enzymes in diverse industries has increased substantially over past decades, creating a well-established and growing global market. Currently, the use of enzymes that work better at ambient or lower temperatures in order to decrease the temperatures of production processes is desirable. There is thus a continuous search for enzymes in cold environments, especially from microbial sources, with amylases, proteases, lipases and, cellulases being the most studied. Other enzymes, such as glucose oxidase (GOD), invertase (Inv), and alkaline phosphatase (ALP), also have a high potential for application, but have been much less studied in microorganisms living in cold-environments. In this work, secretion of these three enzymes by Antarctic yeast species was analyzed, and five, three, and five species were found to produce extracellular GOD, Inv, and ALP, respectively. The major producers of GOD, Inv, and ALP were Goffeauzyma gastrica, Wickerhamomyces anomalus, and Dioszegia sp., respectively, from which the enzymes were purified and characterized. Contrary to what was expected, the highest GOD and Inv activities were found at 64°C and 60°C, respectively, and at 47°C for ALP. However, the three enzymes maintained a significant percentage of activity at lower temperatures, especially ALP that kept a 67 and 43% of activity at 10°C and 4°C, respectively. PMID:29312954

  14. Steady-State ALPS for Real-Valued Problems

    NASA Technical Reports Server (NTRS)

    Hornby, Gregory S.

    2009-01-01

    The two objectives of this paper are to describe a steady-state version of the Age-Layered Population Structure (ALPS) Evolutionary Algorithm (EA) and to compare it against other GAs on real-valued problems. Motivation for this work comes from our previous success in demonstrating that a generational version of ALPS greatly improves search performance on a Genetic Programming problem. In making steady-state ALPS some modifications were made to the method for calculating age and the method for moving individuals up layers. To demonstrate that ALPS works well on real-valued problems we compare it against CMA-ES and Differential Evolution (DE) on five challenging, real-valued functions and on one real-world problem. While CMA-ES and DE outperform ALPS on the two unimodal test functions, ALPS is much better on the three multimodal test problems and on the real-world problem. Further examination shows that, unlike the other GAs, ALPS maintains a genotypically diverse population throughout the entire search process. These findings strongly suggest that the ALPS paradigm is better able to avoid premature convergence then the other GAs.

  15. 13 CFR 120.840 - Accredited Lenders Program (ALP).

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... (ALP). (a) General. Under the ALP program, SBA designates qualified CDCs as ALP CDCs, gives them... approval and servicing actions. (b) Application. A CDC must apply for ALP status to the Lead SBA Office.... (c) Eligibility. In order for a CDC to be eligible to receive ALP status, its application must show...

  16. 13 CFR 120.840 - Accredited Lenders Program (ALP).

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... (ALP). (a) General. Under the ALP program, SBA designates qualified CDCs as ALP CDCs, gives them... approval and servicing actions. (b) Application. A CDC must apply for ALP status to the Lead SBA Office.... (c) Eligibility. In order for a CDC to be eligible to receive ALP status, its application must show...

  17. 13 CFR 120.840 - Accredited Lenders Program (ALP).

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... (ALP). (a) General. Under the ALP program, SBA designates qualified CDCs as ALP CDCs, gives them... approval and servicing actions. (b) Application. A CDC must apply for ALP status to the Lead SBA Office.... (c) Eligibility. In order for a CDC to be eligible to receive ALP status, its application must show...

  18. 13 CFR 120.840 - Accredited Lenders Program (ALP).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... (ALP). (a) General. Under the ALP program, SBA designates qualified CDCs as ALP CDCs, gives them... approval and servicing actions. (b) Application. A CDC must apply for ALP status to the Lead SBA Office.... (c) Eligibility. In order for a CDC to be eligible to receive ALP status, its application must show...

  19. 13 CFR 120.840 - Accredited Lenders Program (ALP).

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... (ALP). (a) General. Under the ALP program, SBA designates qualified CDCs as ALP CDCs, gives them... approval and servicing actions. (b) Application. A CDC must apply for ALP status to the Lead SBA Office.... (c) Eligibility. In order for a CDC to be eligible to receive ALP status, its application must show...

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

    PubMed

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

    2013-03-01

    Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders. Copyright © 2012 Elsevier Inc. All rights reserved.

  1. Dietary phosphate restriction normalizes biochemical and skeletal abnormalities in a murine model of tumoral calcinosis.

    PubMed

    Ichikawa, Shoji; Austin, Anthony M; Gray, Amie K; Allen, Matthew R; Econs, Michael J

    2011-12-01

    Mutations in the GALNT3 gene cause tumoral calcinosis characterized by ectopic calcifications due to persistent hyperphosphatemia. We recently developed Galnt3 knockout mice in a mixed background, which had hyperphosphatemia with increased bone mineral density (BMD) and infertility in males. To test the effect of dietary phosphate intake on their phenotype, Galnt3 knockout mice were generated in the C57BL/6J strain and fed various phosphate diets: 0.1% (low), 0.3% (low normal), 0.6% (normal), and 1.65% (high). Sera were analyzed for calcium, phosphorus, alkaline phosphatase, creatinine, blood urine nitrogen, 1,25-dihydroxyvitamin D, osteocalcin, tartrate-resistant acid phosphatase 5b, and fibroblast growth factor 23 (Fgf23). Femurs were evaluated by dual-energy x-ray absorptiometry, dynamic histomorphometry, and/or microcomputed tomography. Galnt3 knockout mice in C57BL/6J had the same biochemical phenotype observed in our previous study: hyperphosphatemia, inappropriately normal 1,25-dihydroxyvitamin D level, decreased alkaline phosphatase activity, and low intact Fgf23 concentration but high Fgf23 fragments. Skeletal analyses of their femurs revealed significantly high BMD with increased cortical bone area and trabecular bone volume. On all four phosphate diets, Galnt3 knockout mice had consistently higher phosphorus levels and lower alkaline phosphatase and intact Fgf23 concentrations than littermate controls. The low-phosphate diet normalized serum phosphorus, alkaline phosphatase, and areal BMD but failed to correct male infertility in Galnt3 knockout mice. The high-phosphate diet did not increase serum phosphorus concentration in either mutant or control mice due to a compensatory increase in circulating intact Fgf23 levels. In conclusion, dietary phosphate restriction normalizes biochemical and skeletal phenotypes of Galnt3 knockout mice and, thus, can be an effective therapy for tumoral calcinosis.

  2. Dietary Phosphate Restriction Normalizes Biochemical and Skeletal Abnormalities in a Murine Model of Tumoral Calcinosis

    PubMed Central

    Austin, Anthony M.; Gray, Amie K.; Allen, Matthew R.; Econs, Michael J.

    2011-01-01

    Mutations in the GALNT3 gene cause tumoral calcinosis characterized by ectopic calcifications due to persistent hyperphosphatemia. We recently developed Galnt3 knockout mice in a mixed background, which had hyperphosphatemia with increased bone mineral density (BMD) and infertility in males. To test the effect of dietary phosphate intake on their phenotype, Galnt3 knockout mice were generated in the C57BL/6J strain and fed various phosphate diets: 0.1% (low), 0.3% (low normal), 0.6% (normal), and 1.65% (high). Sera were analyzed for calcium, phosphorus, alkaline phosphatase, creatinine, blood urine nitrogen, 1,25-dihydroxyvitamin D, osteocalcin, tartrate-resistant acid phosphatase 5b, and fibroblast growth factor 23 (Fgf23). Femurs were evaluated by dual-energy x-ray absorptiometry, dynamic histomorphometry, and/or microcomputed tomography. Galnt3 knockout mice in C57BL/6J had the same biochemical phenotype observed in our previous study: hyperphosphatemia, inappropriately normal 1,25-dihydroxyvitamin D level, decreased alkaline phosphatase activity, and low intact Fgf23 concentration but high Fgf23 fragments. Skeletal analyses of their femurs revealed significantly high BMD with increased cortical bone area and trabecular bone volume. On all four phosphate diets, Galnt3 knockout mice had consistently higher phosphorus levels and lower alkaline phosphatase and intact Fgf23 concentrations than littermate controls. The low-phosphate diet normalized serum phosphorus, alkaline phosphatase, and areal BMD but failed to correct male infertility in Galnt3 knockout mice. The high-phosphate diet did not increase serum phosphorus concentration in either mutant or control mice due to a compensatory increase in circulating intact Fgf23 levels. In conclusion, dietary phosphate restriction normalizes biochemical and skeletal phenotypes of Galnt3 knockout mice and, thus, can be an effective therapy for tumoral calcinosis. PMID:22009723

  3. Hot Alps (Invited)

    NASA Astrophysics Data System (ADS)

    Speranza, F.; Minelli, L.; Pignatelli, A.; Gilardi, M.

    2013-12-01

    Although it is frequently assumed that crust of Alpine orogens is hot due to the occurrence of thick and young (hence radiogenic) crust, evidence on the thermal ranking of orogens is contradictory. Heat flow measurements from shallow wells (depth ≤ 1 km) in the Alps yield a relatively cold thermal regime of 50-80 mW/m2, but data are likely biased by meteoric cold-water circulation. Here we report on the spectral analysis of the aeromagnetic residuals of northern Italy to derive the Curie point depth (CPD), assumed to represent the 600°C isotherm depth. Airborne magnetics were acquired on whole Italy during the 1970s by the national oil company AGIP (now Eni). Data were gathered by several surveys carried out at 1000-13,300 feet (300-4000 m) altitude, with flight line spacing of 2-10 km. Surveys of the Alps and Po Plain (northern Italy) were obtained both with a line spacing of 5 km (and 5 km tie lines), at an altitude of 4000-5000 and 13,300 feet, respectively. To evaluate CPDs we used the centroid method (routinely adopted in recent CPD studies on East Asia and central-southern Europe) on 72 square windows of 100-110 km edge, with a 50% degree of superposition. CPDs vary between 16 and 38 km (22 km on average) in the Po Plain, located south of the Alps and representing the Adriatic-African foreland area. Conversely, the Alps yield very shallow CPDs, ranging between 6 and 15 km (10 km on average). CPDs fall systematically above local Moho depths, implying that magnetic source bottoms documented in this study do not represent a lithological boundary over non-magnetic peridotitic mantle, but can be safely associated with CPDs and the 600°C isotherm. CPDs from the Po Plain are in rough agreement with reported heat flow values of 25-60 mW/m2, and imply and average thermal conductivity (k) of the Po Plain crust of 1.5 W/m°K, at the lower bound of k values measured and inferred for the crust. Conversely, the average 10 km CPD documented in the Alps translates into

  4. Synthesis and effect of Ce and Mn co-doping on photoluminescence characteristics of Ca6AlP5O20:Eu novel phosphors.

    PubMed

    Shinde, K N; Dhoble, S J

    2013-01-01

    A series of Ca6AlP5O20 doped with rare earths (Eu and Ce) and co-doped (Eu, Ce and Eu,Mn) were prepared by combustion synthesis. Under Hg-free excitation, Ca6AlP5O20:Eu exhibited Eu(2+) (486 nm) emission in the blue region of the spectrum and under near Hg excitation (245 nm), Ca6AlP5O20:Ce phosphor exhibited Ce(3+) emission (357 nm) in the UV range. Photoluminescence (PL) peak intensity increased in Ca6AlP5O20:Eu,Ce and Ca6AlP5O20:Eu, Mn phosphors due to co-activators of Ce(3+) and Mn(2+) ions. As a result, these ions played an important role in PL emission in the present matrix. Ca6AlP5O20:Eu, Ce and Ca6AlP5O20:Eu, Mn phosphors provided energy transfer mechanisms via Ce(3+) → Eu(2+) and Eu(2+) → Mn(2+), respectively. Eu ions acted as activators and Ce ions acted as sensitizers. Ce emission energy was well matched with Eu excitation energy in the case of Ca6AlP5O20:Eu, Ce and Eu ions acted as activators and Mn ions acted as sensitizers in Ca6AlP5O20:Eu, Mn. This study included synthesis of new and efficient phosphate phosphors. The impact of doping and co-doping on photoluminescence properties and energy transfer mechanisms were investigated and we propose a feasible interpretation. Copyright © 2012 John Wiley & Sons, Ltd.

  5. Predictors of severe hemolysis in patients with glucose-6-phosphate dehydrogenase deficiency following exposure to oxidant stresses.

    PubMed

    Al-Sweedan, Suleimman A; Jdaitawi, Hussein; Khriesat, Wadah M; Khader, Yousef Y; Al-Rimawi, Hala S

    2009-01-01

    Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a genetic enzymatic disorder that affects millions of people worldwide, and is a major health problem in Jordan. We studied factors that may predict severe hemolysis in children with G6PD deficiency. We reviewed the records of patients with low G6PD activity admitted to a teaching hospital be- tween 1996 to 2007. We collected demographic data, details of sign and symptoms, history and type of fava bean ingestion, blood and Rh group, history of neonatal jaundice, history and type of drug use, abdominal pain at admission and the results of tests for hemoglobin, white blood cells (WBC), and hepatic function. We classified patients into mild and severe groups based on hemoglobin levels at admission. Of 428 children with G6PD deficiency, 79 (18%) were severe cases and 349 (82%) patients with mild disease. There were no statistically significant differences in most factors between the two groups. Factors that achieved statistical significance for severe hemolysis included younger age (P<.05), male gender (P<.05), higher alkaline phosphatase (ALP) (P<.05), presence of fever at admission (P<.01), presence of vomiting during the at- tack (P=.006), and a negative family history for G6PD deficiency (P=.005). Severe hemolysis can be predicted during hemolytic episodes in children with low G6PD by young age, male gender, a negative family history of G6PD deficiency, the presence of fever and vomiting and a high ALP.

  6. Maxillary sinus floor elevation using a tissue-engineered bone with calcium-magnesium phosphate cement and bone marrow stromal cells in rabbits.

    PubMed

    Zeng, Deliang; Xia, Lunguo; Zhang, Wenjie; Huang, Hui; Wei, Bin; Huang, Qingfeng; Wei, Jie; Liu, Changsheng; Jiang, Xinquan

    2012-04-01

    The objective of this study was to assess the effects of maxillary sinus floor elevation with a tissue-engineered bone constructed with bone marrow stromal cells (bMSCs) and calcium-magnesium phosphate cement (CMPC) material. The calcium (Ca), magnesium (Mg), and phosphorus (P) ions released from calcium phosphate cement (CPC), magnesium phosphate cement (MPC), and CMPC were detected by inductively coupled plasma atomic emission spectroscopy (ICP-AES), and the proliferation and osteogenic differentiation of bMSCs seeded on CPC, MPC, and CMPC or cultured in CPC, MPC, and CMPC extracts were measured by MTT analysis, alkaline phosphatase (ALP) activity assay, alizarin red mineralization assay, and real-time PCR analysis of the osteogenic genes ALP and osteocalcin (OCN). Finally, bMSCs were combined with CPC, MPC, and CMPC and used for maxillary sinus floor elevation in rabbits, while CPC, MPC, or CMPC without cells served as control groups. The new bone formation in each group was detected by histological finding and fluorochrome labeling at weeks 2 and 8 after surgical operation. It was observed that the Ca ion concentrations of the CMPC and CPC scaffolds was significantly higher than that of the MPC scaffold, while the Mg ions concentration of CMPC and MPC was significantly higher than that of CPC. The bMSCs seeded on CMPC and MPC or cultured in their extracts proliferated more quickly than the cells seeded on CPC or cultured in its extract, respectively. The osteogenic differentiation of bMSCs seeded on CMPC and CPC or cultured in the corresponding extracts was significantly enhanced compared to that of bMSCs seeded on MPC or cultured in its extract; however, there was no significant difference between CMPC and CPC. As for maxillary sinus floor elevation in vivo, CMPC could promote more new bone formation and mineralization compared to CPC and MPC, while the addition of bMSCs could further enhance its new bone formation ability significantly. Our data suggest that

  7. Osteogenic differentiation of immature osteoblasts: Interplay of cell culture media and supplements.

    PubMed

    Brauer, A; Pohlemann, T; Metzger, W

    2016-01-01

    Differentiation of immature osteoblasts to mature osteoblasts in vitro initially was induced by supplementing the medium with β-gylcerophosphate and dexamethasone. Later, ascorbic acid, vitamin D3, vitamin K3 and TGFβ1 were used in varying concentrations as supplements to generate a mature osteoblast phenotype. We tested the effects of several combinations of cell culture media, seeding protocols and osteogenic supplements on osteogenic differentiation of human primary osteoblasts. Osteogenic differentiation was analyzed by staining alkaline phosphatase (ALP) with 5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium (BCIP/NBT) and by von Kossa staining of deposited calcium phosphate. The combinations of culture media and supplements significantly influenced osteogenic differentiation, but the seeding protocol did not. Staining of ALP and calcium phosphate could be achieved only if our own mix of osteogenic supplements was used in combination with Dulbecco's modified Eagle medium or if a commercial mix of osteogenic supplements was used in combination with osteoblast growth medium. Especially for von Kossa, we observed great variations in the staining intensity. Because osteogenic differentiation is a complex process, the origin of the osteoblasts, cell culture media and osteogenic supplements should be established by preliminary experiments to achieve optimal differentiation. Staining of ALP or deposited calcium phosphate should be supplemented with qRT-PCR studies to learn more about the influence of specific supplements on osteogenic markers.

  8. Serum bone alkaline phosphatase and calcaneus bone density predict fractures: a prospective study.

    PubMed

    Ross, P D; Kress, B C; Parson, R E; Wasnich, R D; Armour, K A; Mizrahi, I A

    2000-01-01

    The aim of this study was to assess the ability of serum bone-specific alkaline phosphatase (bone ALP), creatinine-corrected urinary collagen crosslinks (CTx) and calcaneus bone mineral density (BMD) to identify postmenopausal women who have an increased risk of osteoporotic fractures. Calcaneus BMD and biochemical markers of bone turnover (serum bone ALP and urinary CTx) were measured in 512 community-dwelling postmenopausal women (mean age at baseline 69 years) participating in the Hawaii Osteoporosis Study. New spine and nonspine fractures subsequent to the BMD and biochemical bone markers measurements were recorded over an average of 2.7 years. Lateral spinal radiographs were used to identify spine fractures. Nonspine fractures were identified by self-report at the time of each examination. During the 2.7-year follow-up, at least one osteoporotic fracture occurred in 55 (10.7%) of the 512 women. Mean baseline serum bone ALP and urinary CTx were significantly higher among women who experienced an osteoporotic fracture compared with those women who did not fracture. In separate age-adjusted logistic regression models, serum bone ALP, urinary CTx and calcaneus BMD were each significantly associated with new fractures (odds ratios of 1.53, 1.54 and 1.61 per SD, respectively). Multiple variable logistic regression analysis identified BMD and serum bone ALP as significant predictors of fracture (p = 0.002 and 0.017, respectively). The results from this investigation indicate that increased bone turnover is significantly associated with an increased risk of osteoporotic fracture in postmenopausal women. This association is similar in magnitude and independent of that observed for BMD.

  9. Delivering MC3T3-E1 cells into injectable calcium phosphate cement through alginate-chitosan microcapsules for bone tissue engineering*

    PubMed Central

    Qiao, Peng-yan; Li, Fang-fang; Dong, Li-min; Xu, Tao; Xie, Qiu-fei

    2014-01-01

    Objective: To deliver cells deep into injectable calcium phosphate cement (CPC) through alginate-chitosan (AC) microcapsules and investigate the biological behavior of the cells released from microcapsules into the CPC. Methods: Mouse osteoblastic MC3T3-E1 cells were embedded in alginate and AC microcapsules using an electrostatic droplet generator. The two types of cell-encapsulating microcapsules were then mixed with a CPC paste. MC3T3-E1 cell viability was investigated using a Wst-8 kit, and osteogenic differentiation was demonstrated by an alkaline phosphatase (ALP) activity assay. Cell attachment in CPC was observed by an environment scanning electron microscopy. Results: Both alginate and AC microcapsules were able to release the encapsulated MC3T3-E1 cells when mixed with CPC paste. The released cells attached to the setting CPC scaffolds, survived, differentiated, and formed mineralized nodules. Cells grew in the pores concomitantly created by the AC microcapsules in situ within the CPC. At Day 21, cellular ALP activity in the AC group was approximately four times that at Day 7 and exceeded that of the alginate microcapsule group (P<0.05). Pores formed by the AC microcapsules had a diameter of several hundred microns and were spherical compared with those formed by alginate microcapsules. Conclusions: AC microcapsule is a promising carrier to release seeding cells deep into an injectable CPC scaffold for bone engineering. PMID:24711359

  10. Effect of carbonated hydroxyapatite incorporated advanced platelet rich fibrin intrasulcular injection on the alkaline phosphatase level during orthodontic relapse

    NASA Astrophysics Data System (ADS)

    Alhasyimi, Ananto Ali; Pudyani, Pinandi Sri; Asmara, Widya; Ana, Ika Dewi

    2018-02-01

    Nowadays, relapse in orthodontic treatment is considered very important because of high incidence of relapse after the treatment. Alkaline phosphatase (ALP) as a biomarker of bone formation will decrease in compression sites during relapse after orthodontic tooth movement. In this situation, manipulating alveolar bone remodeling to increase ALP level is considered one of the new strategies to prevent relapse properly. In the field of tissue engineering, in this study, carbonated hydroxyapatite (CHA) is expected to have the ability to incorporate advanced platelet rich fibrin (aPRF). Next, CHA will retain the aPRF containing various growth factors (GF) until it reaches into a specific targeted area, gradually degraded, and deliver the GF in a controlled manner to prevent relapse. Here, gingival crevicular fluid (GCF) of 45 samples (n=45) were collected and levels of ALP were analyzed using UV-Vis 6300 Spectrophotometer at 405 nm wavelength. We found that there is a significant difference of ALP levels (p<0.05) in GCF between treatments and control groups. ALP level was elevated significantly in CHA and CHA-aPRF groups at days 7 and 14 after debonding compared with the control groups. The peak level of ALP was observed at days 14 after debonding in groups C (0.789 ± 0.039 U/mg). Therefore, it can be concluded that the application of hydrogel CHA with controlled release manner incorporated aPRF enhances bone regeneration by increasing ALP level.

  11. Metabolic effect of alkaline additives and guanosine/gluconate in storage solutions for red blood cells.

    PubMed

    D'Alessandro, Angelo; Reisz, Julie A; Culp-Hill, Rachel; Korsten, Herbert; van Bruggen, Robin; de Korte, Dirk

    2018-04-06

    Over a century of advancements in the field of additive solutions for red blood cell (RBC) storage has made transfusion therapy a safe and effective practice for millions of recipients worldwide. Still, storage in the blood bank results in the progressive accumulation of metabolic alterations, a phenomenon that is mitigated by storage in novel storage additives, such as alkaline additive solutions. While novel alkaline additive formulations have been proposed, no metabolomics characterization has been performed to date. We performed UHPLC-MS metabolomics analyses of red blood cells stored in SAGM (standard additive in Europe), (PAGGSM), or alkaline additives SOLX, E-SOL 5 and PAG3M for either 1, 21, 35 (end of shelf-life in the Netherlands), or 56 days. Alkaline additives (especially PAG3M) better preserved 2,3-diphosphoglycerate and adenosine triphosphate (ATP). Deaminated purines such as hypoxanthine were predictive of hemolysis and morphological alterations. Guanosine supplementation in PAGGSM and PAG3M fueled ATP generation by feeding into the nonoxidative pentose phosphate pathway via phosphoribolysis. Decreased urate to hypoxanthine ratios were observed in alkaline additives, suggestive of decreased generation of urate and hydrogen peroxide. Despite the many benefits observed in purine and redox metabolism, alkaline additives did not prevent accumulation of free fatty acids and oxidized byproducts, opening a window for future alkaline formulations including (lipophilic) antioxidants. Alkalinization via different strategies (replacement of chloride anions with either high bicarbonate, high citrate/phosphate, or membrane impermeant gluconate) results in different metabolic outcomes, which are superior to current canonical additives in all cases. © 2018 AABB.

  12. Alkaline phosphatase binds tenaciously to titanium; implications for biological surface evaluation following bone implant retrieval.

    PubMed

    Mansell, J P; Shiel, A I; Harwood, C; Stephens, D

    2017-07-01

    Enhancing the performance and longevity of titanium (Ti) implants continues to be a significant developmental theme in contemporary biomaterials design. Our specific focus pertains to the surface functionalisation of Ti using the bioactive lipid, lysophosphatidic acid (LPA) and certain phosphatase-resistant analogues of LPA. Coating survivorship to a plethora of testing regimens is required to align with due regulatory process before novel biomaterials can enter clinical trials. One of the key acceptance criteria is coating retention to the physical stresses experienced during implantation. In assessing coating stability to insertion into porcine bone we found that a subsequent in vitro assessment to confirm coating persistence was masked by abundant alkaline phosphatase (ALP) contamination adsorbed to the metal surface. Herein we report that ALP can bind to Ti in a matter of minutes by simply immersing Ti samples in aqueous solutions of the enzyme. We strongly discourage the in vitro monitoring of osteoblast and stromal cell ALP expression when assessing bioactive coating survivorship following Ti implant retrieval form native bone tissue. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Simultaneous removal of ammonium and phosphate by alkaline-activated and lanthanum-impregnated zeolite.

    PubMed

    He, Yinhai; Lin, Hai; Dong, Yingbo; Liu, Quanli; Wang, Liang

    2016-12-01

    Simultaneous ammonium and phosphate removal characteristics and mechanism, as well as the major influencing factors, such as pH, temperature and co-existing ions, onto NaOH-activated and lanthanum-impregnated zeolite (NLZ) were investigated. The phosphate adsorption increases from 0.2 mg g -1 for natural zeolite up to 8.96 mg g -1 for NLZ, while only a slight decrease on the ammonium adsorption capacity from 23.9 mg g -1 for NaOH-activated zeolite to 21.2 mg g -1 for NLZ was observed. The ammonium and phosphate adsorption showed little pH dependence in the range from pH 3 to 7, while it decreased sharply with the pH increased above pH 7. Adsorption of ammonium and phosphate could be well described by the pseudo-second-order model and the process was mainly governed by intra-particle diffusion. The Langmuir and Freundlich model can be acceptably applied to fit the experimental data, which suggested that adsorption was caused by both the monolayer and homogeneous coverage at specific and equal affinity sites available NLZ. The underlying mechanism for the specific adsorption of phosphate by NLZ was revealed with the aid of SEM-EDS, XPS, and FTIR analysis, and the formation of (LaO)(OH)PO 2 was verified to be the dominant pathway for selective phosphate adsorption by lanthanum-impregnated zeolite. While the removal mechanism of ammonium could be well interpreted by SEM-EDS, FTIR and ICP analysis, and ion-exchange was expected to be the main removal process for ammonium. The results indicate that NLZ could efficiently and simultaneously remove low concentration of ammonium and phosphate from contaminated waters. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Apatite mineralization in elasmobranch skeletons via a polyphosphate intermediate

    NASA Astrophysics Data System (ADS)

    Omelon, Sidney; Lacroix, Nicolas; Lildhar, Levannia; Variola, Fabio; Dean, Mason

    2014-05-01

    All vertebrate skeletons are stiffened with apatite, a calcium phosphate mineral. Control of apatite mineralization is essential to the growth and repair of the biology of these skeletons, ensuring that apatite is deposited in the correct tissue location at the desired time. The mechanism of this biochemical control remains debated, but must involve increasing the localized apatite saturation state. It was theorized in 1923 that alkaline phosphatase (ALP) activity provides this control mechanism by increasing the inorganic phosphate (Pi) concentration via dephosphorylation of phosphorylated molecules. The ALP substrate for biological apatite is not known. We propose that polyphosphates (polyPs) produced by mitochondria may be the substrate for biological apatite formation by ALP activity. PolyPs (PO3-)n, also known as condensed phosphates, represent a concentrated, bioavailable Pi-storage strategy. Mitochondria import Pi and synthesize phosphate polymers through an unknown biochemical mechanism. When chelated with calcium and/or other cations, the effective P-concentration of these neutrally charged, amorphous, polyP species can be very high (~ 0.5 M), without inducing phosphate mineral crystallization. This P-concentration in the low Pi-concentration biological environment offers a method of concentrating P well above an apatite supersaturation required for nucleation. Bone is the most studied mineralized skeletal tissue. However, locating and analyzing active mineralizing areas is challenging. We studied calcified cartilage skeletons of elasmobranch fishes (sharks, stingrays and relatives) to analyse the phosphate chemistry in this continually mineralizing skeleton. Although the majority of the elasmobranch skeleton is unmineralized cartilage, it is wrapped in an outer layer of mineralized tissue comprised of small tiles called tesserae. These calcified tesserae continually grow through the formation of new mineral on their borders. Co-localization of ALP and

  15. P wave anisotropic tomography of the Alps

    NASA Astrophysics Data System (ADS)

    Hua, Yuanyuan; Zhao, Dapeng; Xu, Yixian

    2017-06-01

    The first tomographic images of P wave azimuthal and radial anisotropies in the crust and upper mantle beneath the Alps are determined by joint inversions of arrival time data of local earthquakes and teleseismic events. Our results show the south dipping European plate with a high-velocity (high-V) anomaly beneath the western central Alps and the north dipping Adriatic plate with a high-V anomaly beneath the Eastern Alps, indicating that the subduction polarity changes along the strike of the Alps. The P wave azimuthal anisotropy is characterized by mountain chain-parallel fast-velocity directions (FVDs) in the western central Alps and NE-SW FVDs in the Eastern Alps, which may be caused by mantle flow induced by the slab subductions. Our results reveal a negative radial anisotropy (i.e., Vph < Vpv) within the subducting slabs and a positive radial anisotropy (i.e., Vph > Vpv) in the low-velocity mantle wedge, which may reflect the subvertical plate subduction and its induced mantle flow. The results of anisotropic tomography provide important new information on the complex mantle structure and dynamics of the Alps and adjacent regions.

  16. Analysis of the ectoenzymes ADA, ALP, ENPP1, and ENPP3, in the contents of ovarian endometriomas as candidate biomarkers of endometriosis.

    PubMed

    Trapero, Carla; Jover, Lluis; Fernández-Montolí, Maria Eulàlia; García-Tejedor, Amparo; Vidal, August; Gómez de Aranda, Inmaculada; Ponce, Jordi; Matias-Guiu, Xavier; Martín-Satué, Mireia

    2018-02-01

    The diagnosis of endometriosis, a prevalent chronic disease with a strong inflammatory component, is usually delayed due to the lack of noninvasive diagnostic tests. Purinergic signaling, a key cell pathway, is altered in many inflammatory disorders. The aim of the present work was to evaluate the levels of adenosine deaminase (ADA), alkaline phosphatase (ALP), ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), and ENPP3, elements of purinergic signaling, as biomarker candidates for endometriosis. A case-control comparative study was conducted to determine ADA, ALP, ENPP1 and ENPP3 levels in echo-guided aspirated fluids of endometriomas (case group) and simple ovarian cysts (control group) using the ELISA technique. Adenosine deaminase, ALP, ENPP1, and ENPP3 were present and quantifiable in the contents of endometriomas and simple cysts. There were significant differences in ADA and ENPP1 levels in endometriomas in comparison with simple cysts (2787 U/L and 103.9 ng/mL more in endometriomas, for ADA and ENPP1, respectively). Comparisons of ALP and ENPP3 levels between the two groups did not reveal significant differences. The ectoenzymes ADA and ENPP1 are biomarker candidates for endometriosis. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Beta-tricalcium phosphate granules improve osteogenesis in vitro and establish innovative osteo-regenerators for bone tissue engineering in vivo.

    PubMed

    Gao, Peng; Zhang, Haoqiang; Liu, Yun; Fan, Bo; Li, Xiaokang; Xiao, Xin; Lan, Pingheng; Li, Minghui; Geng, Lei; Liu, Dong; Yuan, Yulin; Lian, Qin; Lu, Jianxi; Guo, Zheng; Wang, Zhen

    2016-03-22

    The drawbacks of traditional bone-defect treatments have prompted the exploration of bone tissue engineering. This study aimed to explore suitable β-tricalcium phosphate (β-TCP) granules for bone regeneration and identify an efficient method to establish β-TCP-based osteo-regenerators. β-TCP granules with diameters of 1 mm and 1-2.5 mm were evaluated in vitro. The β-TCP granules with superior osteogenic properties were used to establish in vivo bioreactors, referred to as osteo-regenerators, which were fabricated using two different methods. Improved proliferation of bone mesenchymal stem cells (BMSCs), glucose consumption and ALP activity were observed for 1-2.5 mm β-TCP compared with 1-mm granules (P < 0.05). In addition, BMSCs incubated with 1-2.5 mm β-TCP expressed significantly higher levels of the genes for runt-related transcription factor-2, alkaline phosphatase, osteocalcin, osteopontin, and collagen type-1 and the osteogenesis-related proteins alkaline phosphatase, collagen type-1 and runt-related transcription factor-2 compared with BMSCs incubated with 1 mm β-TCP (P < 0.05). Fluorochrome labelling, micro-computed tomography and histological staining analyses indicated that the osteo-regenerator with two holes perforating the femur promoted significantly greater bone regeneration compared with the osteo-regenerator with a periosteum incision (P < 0.05). This study provides an alternative to biofunctionalized bioreactors that exhibits improved osteogenesis.

  18. Beta-tricalcium phosphate granules improve osteogenesis in vitro and establish innovative osteo-regenerators for bone tissue engineering in vivo

    PubMed Central

    Gao, Peng; Zhang, Haoqiang; Liu, Yun; Fan, Bo; Li, Xiaokang; Xiao, Xin; Lan, Pingheng; Li, Minghui; Geng, Lei; Liu, Dong; Yuan, Yulin; Lian, Qin; Lu, Jianxi; Guo, Zheng; Wang, Zhen

    2016-01-01

    The drawbacks of traditional bone-defect treatments have prompted the exploration of bone tissue engineering. This study aimed to explore suitable β-tricalcium phosphate (β-TCP) granules for bone regeneration and identify an efficient method to establish β-TCP-based osteo-regenerators. β-TCP granules with diameters of 1 mm and 1–2.5 mm were evaluated in vitro. The β-TCP granules with superior osteogenic properties were used to establish in vivo bioreactors, referred to as osteo-regenerators, which were fabricated using two different methods. Improved proliferation of bone mesenchymal stem cells (BMSCs), glucose consumption and ALP activity were observed for 1–2.5 mm β-TCP compared with 1-mm granules (P < 0.05). In addition, BMSCs incubated with 1–2.5 mm β-TCP expressed significantly higher levels of the genes for runt-related transcription factor-2, alkaline phosphatase, osteocalcin, osteopontin, and collagen type-1 and the osteogenesis-related proteins alkaline phosphatase, collagen type-1 and runt-related transcription factor-2 compared with BMSCs incubated with 1 mm β-TCP (P < 0.05). Fluorochrome labelling, micro-computed tomography and histological staining analyses indicated that the osteo-regenerator with two holes perforating the femur promoted significantly greater bone regeneration compared with the osteo-regenerator with a periosteum incision (P < 0.05). This study provides an alternative to biofunctionalized bioreactors that exhibits improved osteogenesis. PMID:27000963

  19. Alkaline Hydrothermal Vents as Electrochemical Reactors Driving an Auto-Trophic Origin of Life

    NASA Astrophysics Data System (ADS)

    Camprubi, E.; Lane, N.

    2017-07-01

    We report the reduction of CO2 to formaldehyde under simulated alkaline hydrothermal conditions. Formaldehyde is transformed into relevant sugars via the formose reaction. Acetyl phosphate can also be synthesised and phosphorylates organic molecules.

  20. Effect of biomimetic zinc-containing tricalcium phosphate (Zn-TCP) on the growth and osteogenic differentiation of mesenchymal stem cells.

    PubMed

    Chou, Joshua; Hao, Jia; Hatoyama, Hirokazu; Ben-Nissan, Besim; Milthorpe, Bruce; Otsuka, Makoto

    2015-07-01

    Several studies have shown the effectiveness of zinc-tricalcium phosphate (Zn-TCP) for bone tissue engineering. In this study, marine calcareous foraminifera possessing uniform pore size distribution were hydrothermally converted to Zn-TCP. The ability of a scaffold to combine effectively with mesenchymal stem cells (MSCs) is a key tissue-engineering aim. In order to demonstrate the osteogenic ability of MSCs with Zn-TCP, the scaffolds were cultured in an osteogenic induction medium to elicit an osteoblastic response. The physicochemical properties of Zn-TCP were characterized by XRD, FT-IR and ICP-MS. MSCs were aspirated from rat femurs and cultured for 3 days before indirectly placing four samples into each respective well. After culture for 7, 10 and 14 days, osteoblastic differentiation was evaluated using alizarin red S stain, measurement of alkaline phosphatase (ALP) levels, cell numbers and cell viability. XRD and FT-IR patterns both showed the replacement of CO(3)(2-) with PO(4)(3-). Chemical analysis showed zinc incorporation of 5 mol%. Significant increases in cell numbers were observed at 10 and 14 days in the Zn-TCP group, while maintaining high levels of cell viability (> 90%). ALP activity in the Zn-TCP group was statistically higher at 10 days. Alizarin red S staining also showed significantly higher levels of calcium mineralization in Zn-TCP compared with the control groups. This study showed that MSCs in the presence of biomimetically derived Zn-TCP can accelerate their differentiation to osteoblasts and could potentially be useful as a scaffold for bone tissue engineering. Copyright © 2014 John Wiley & Sons, Ltd.

  1. Ratiometric detection of copper ions and alkaline phosphatase activity based on semiconducting polymer dots assembled with rhodamine B hydrazide.

    PubMed

    Sun, Junyong; Mei, Han; Gao, Feng

    2017-05-15

    The rational surface functionalization of semiconducting polymer dots (Pdots) has attracted much attention to extend their applications in fabricating chemo/biosensing platform. In this study, a novel ratiometric fluorescent sensing platform using functionalized Pdots as probes for fluorescence signal transmission has been designed for sensing Cu(Ⅱ) and activity of alkaline phosphatase (ALP) with high selectivity and enhanced sensitivity. The highly fluorescent Pdots were firstly prepared with Poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-benzo-{2,1',3}-thiadiazole)] (PFBT) via nanoprecipitation method, and then assembled with non-fluorescent rhodamine B hydrazide (RB-hy), which shows special binding activity to Cu(Ⅱ), through adsorption process to obtain functionalized nanohybrids, Pdots@RB-hy. As thus, a FRET donors/acceptors pair, in which PFBT Pdots act as energy donors while RB-hy-Cu(II) complexes act as energy acceptors were constructed. On the basis of the varies in fluorescence intensities of donors/acceptors in the presence of different amounts of Cu(II), a ratiometric method for sensing Cu(II) has been proposed. The proposed ratiometric Cu(II) sensor shows a good linear detection range from 0.05 to 5μM with a detection limit of 15nM. Furthermore, using the Pdots@RB-hy-Cu(II) system as signal transducer, a ratiometric sensing for alkaline phosphatase (ALP) activity has also been established with pyrophosphate (PPi) as substrates. The constructed ratiometric sensor of ALP activity displays a linear detection range from 0.005 to 15UL -1 with a detection limit of 0.0018UL -1 . The sensor was further successfully used for ALP activity detection in human serum with satisfactory results. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    PubMed

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

    2016-01-01

    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. 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. 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. Independent variables for measurements over time were analyzed by using Wilcoxon signed rank test. 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. 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.

  3. Apoptosis may determine the release of skeletal alkaline phosphatase activity from human osteoblast-line cells.

    PubMed

    Farley, J R; Stilt-Coffing, B

    2001-01-01

    Although quantitative measurement of skeletal alkaline phosphatase (sALP) activity in serum can provide an index of the rate of bone formation, the metabolic process that determines the release of sALP - from the surface of osteoblasts, into circulation-is unknown. The current studies were intended to examine the hypothesis that the release of sALP from human osteoblasts is a consequence of apoptotic cell death. We measured the release of sALP activity from human osteosarcoma (SaOS-2) cells and normal human bone cells, under basal conditions and in response to agents that increased apoptosis (TNF-a, okadiac acid) and agents that inhibit apoptosis (IGF-I, calpain, and caspase inhibitors). Apoptosis was determined by the presence of nucleosomes (histone-associated DNA) in the cytoplasm of the cells by using a commercial kit. The results of these studies showed that TNF-a and okadiac acid caused dose- and time-dependent increases in apoptosis in the SaOS-2 cells (r = 0.78 for doses of TNF-a and r = 0.93 for doses of okadiac acid, P <0.005 for each), with associated decreases in cell layer protein (P <0.05 for each) and concomitant increases in the release of sALP activity (e.g., r = 0.89 for TNF-a and r = 0.75 for okadiac acid, P <0.001 for each). In contrast, caspase and calpain inhibitors reduced apoptosis, increased cell layer protein, and decreased the release of sALP activity (P <0.05 for each). Exposure to IGF-I also decreased apoptosis, in a time- and dose-dependent manner (e.g., r = 0.93, P <0.001 for IGF-I doses), with associated proportional effects to increase cell layer protein (P <0.001) and decrease the release of sALP activity (P <0.001). IGF-I also inhibited the actions of TNF-a and okadiac acid to increase apoptosis and sALP release. The associations between apoptosis and sALP release were not unique to osteosarcoma (i.e., SaOS-2) cells, but also seen with osteoblast-line cells derived from normal human bone. Together, these data demonstrate that the

  4. Heterologous expression and structure-function relationship of low-temperature and alkaline active protease from Acinetobacter sp. IHB B 5011(MN12).

    PubMed

    Salwan, Richa; Sharma, Vivek; Pal, Mohinder; Kasana, Ramesh Chand; Yadav, Sudesh Kumar; Gulati, Arvind

    2018-02-01

    The gene encoding protease from Acinetobacter sp. IHB B 5011(MN12) was cloned and expressed in Escherichia coli BL21(DE3). The nucleotide sequence revealed 1323bp ORF encoding 441 amino acids protein with molecular weight 47.2kDa. The phylogenetic analysis showed clustering of Alp protease with subtilisin-like serine proteases of S8 family. The amino acid sequence was comprised of N-terminal signal peptide 1-21 amino acids, pre-peptide 22-143 amino acids, peptidase S8 domain 144-434 amino acids, and pro-peptide 435-441 amino acids at C-terminus. Three constructs with signal peptide pET-Alp, without signal peptide pET-Alp1 and peptidase S8 domain pET-Alp2 were prepared for expression in E. coli BL21(DE3). The recombinant proteins Alp1 and Alp2 expressed as inclusion bodies showed ∼50kDa and ∼40kDa bands, respectively. The pre-propeptide ∼11kDa removed from Alp1 resulted in mature protein of ∼35kDa with 1738Umg -1 specific activity. The recombinant protease was optimally active at 40°C and pH 9, and stable over 10-70°C and 6-12pH. The activity at low-temperature and alkaline pH was supported by high R/(R+K) ratio, more glycine, less proline, negatively charged amino acids, less salt bridges and longer loops. These properties suggested the suitability of Alp as additive in the laundry. Copyright © 2017. Published by Elsevier B.V.

  5. Total and bone-specific alkaline phosphatase are associated with bone mineral density over time in end-stage renal disease patients starting dialysis.

    PubMed

    Bergman, Annelie; Qureshi, Abdul Rashid; Haarhaus, Mathias; Lindholm, Bengt; Barany, Peter; Heimburger, Olof; Stenvinkel, Peter; Anderstam, Björn

    2017-04-01

    Alkaline phosphatase (ALP) and bone-specific ALP (BALP) are implicated in the abnormal skeletal mineralization and accelerated vascular calcification in chronic kidney disease (CKD) patients. Whereas ALP and BALP may predict mortality in CKD, BALP is reported to have higher sensitivity and specificity than total ALP in reflecting histological alterations in bone; however, results on their associations with bone mineral density (BMD) are inconsistent. Here we evaluated associations of total ALP and BALP with BMD during up to 24 months in end-stage renal disease (ESRD) patients. In this longitudinal study, 194 ESRD patients (median age 57 years, 66 % male, 32 % diabetes mellitus, mean body mass index 24.8 kg/m 2 ) underwent measurements of total ALP and BALP and total and regional body BMD (by dual-energy X-ray absorptiometry) at dialysis initiation (n = 194), and after 12 (n = 98) and 24 months (n = 40) on dialysis. At baseline, patients had median total ALP 65.4 (43.3-126.4) U/l, BALP 13.5 (7.1-27.3) µg/l and BMD 1.14 (0.97-1.31) g/cm 2 . During the study period, serum concentrations of ALP and BALP increased significantly (p < 0.001), whereas total and regional BMD remained stable. BMD correlated inversely with total ALP (rho = -0.20, p = 0.005) and BALP (rho = -0.30, p < 0.001) at baseline, and correlations were similar also at 12 and 24 months. ALP and BALP are equally accurate albeit weak predictors of BMD in ESRD patients, both at baseline and longitudinally. The dissociation between stable BMD and increasing ALP and BALP may possibly reflect increased soft tissue calcifications with time on dialysis.

  6. Characterization and in vitro biological evaluation of mineral/osteogenic growth peptide nanocomposites synthesized biomimetically on titanium

    NASA Astrophysics Data System (ADS)

    Chen, Cen; Kong, Xiangdong; Zhang, Sheng-Min; Lee, In-Seop

    2015-04-01

    Nanocomposite layers of mineral/osteogenic growth peptide (OGP) were synthesized on calcium phosphate coated titanium substrates by immersing in calcium-phosphate buffer solution containing OGP. Peptide incorporated mineral was characterized by determining quantity loaded, effects on mineral morphology and structure. Also, the biological activity was investigated by cell adhesion, proliferation assay, and measurement of alkaline phosphatase (ALP) activity. X-ray photoelectron spectroscopy (XPS) and micro-bicinchoninic acid (BCA) assay revealed that OGP was successfully incorporated with mineral and the amount was increased with immersion time. Incorporated OGP changed the mineral morphology from sharp plate-like shape to more rounded one, and the octacalcium phosphate structure of the mineral was gradually transformed into apatite. With confocal microscopy to examine the incorporation of fluorescently labeled peptide, OGP was evenly distributed throughout mineral layers. Mineral/OGP nanocomposites promoted cell adhesion and proliferation, and also increased ALP activity of mesenchymal stem cells (MSCs). Results presented here indicated that the mineral/OGP nanocomposites formed on titanium substrates had the potential for applications in dental implants.

  7. Cloning, characterization, expression and antifungal activity of an alkaline serine protease of Aureobasidium pullulans PL5 involved in the biological control of postharvest pathogens.

    PubMed

    Zhang, Dianpeng; Spadaro, Davide; Valente, Silvia; Garibaldi, Angelo; Gullino, Maria Lodovica

    2012-02-15

    An alkaline protease gene was amplified from genomic DNA and cDNA of the antagonistic yeast-like fungus Aureobasidium pullulans PL5, a biocontrol agent effective against Monilinia laxa on stone fruit and Botrytis cinerea and Penicillium expansum on pome fruits. An open reading frame of 1248 bp encoding a 415-amino acid (aa) protein with a calculated molecular weight (M(r)) of 42.9 kDa and an isoelectric point (pI) of 4.5 was characterized. The cDNAALP5 gene had an 18-amino acid signal peptide, one N-gylcosylation, one histidine active site, and one serine active site. The ALP5 gene with a M(r) of 1351 bp contained two introns. One intron was of 54 bp, while the other was of 50 bp. Protein BLAST and phylogenetic tree analysis of the deduced amino sequences from the cDNAALP5 gene showed that the encoded protein had 100% homology to a protease enzyme (ALP2) of a sea strain of A. pullulans, suggesting that the protein ALP5 was an alkaline serine protease. Expression of ALP5 in Escherichia coli BL21 (DE3), followed by identification with Western-blotting, purification with Ni-NTA and analysis of enzymatic activity, yielded an homogeneous recombinant ALP5 which hydrolysed the substrate casein and inhibited the mycelial growth of the pathogens. At its optimal pH of 10.0 and reaction temperature of 50°C, the recombinant protease exhibited the highest activity towards the substrate casein, though the highest stability was at lower temperatures and pH between 7.0 and 9.0. This study provided the direct evidence that extracellular proteases secreted by the antagonist A. pullulans PL5 played a role in the biocontrol activities against some postharvest pathogens of apple and peach. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. Copper(II) complexes of methimazole, an anti Grave's disease drug. Synthesis, characterization and its potential biological behavior as alkaline phosphatase inhibitor.

    PubMed

    Urquiza, Nora M; Manca, Silvia G; Moyano, María A; Dellmans, Raquel Arrieta; Lezama, Luis; Rojo, Teófilo; Naso, Luciana G; Williams, Patricia A M; Ferrer, Evelina G

    2010-04-01

    Methimazole (MeimzH) is an anti-thyroid drug and the first choice for patients with Grave's disease. Two new copper(II) complexes of this drug: [Cu(MeimzH)(2)(NO(3))(2)]*0.5H(2)O and [Cu(MeimzH)(2)(H(2)O)(2)](NO(3))(2)*H(2)O were synthesized and characterized by elemental analysis, dissolution behavior, thermogravimetric analysis and UV-vis, diffuse reflectance, FTIR and EPR spectroscopies. As it is known that copper(II) cation can act as an inhibitor of alkaline phosphatase (ALP), the inhibitory effect of methimazole and its copper(II) complexes on ALP activity has also been investigated.

  9. Phosphate fertilizer impacts on glyphosate sorption by soil.

    PubMed

    Munira, Sirajum; Farenhorst, Annemieke; Flaten, Don; Grant, Cynthia

    2016-06-01

    This research examined the impact of field-aged phosphate and cadmium (Cd) concentrations, and fresh phosphate co-applications, on glyphosate sorption by soil. Soil samples were collected in 2013 from research plots that had received, from 2002 to 2009, annual applications of mono ammonium phosphate (MAP) at 20, 40 and 80 kg P ha(-1) and from products containing 0.4, 70 or 210 mg Cd kg(-1) as an impurity. A series of batch equilibrium experiments were carried out to quantify the glyphosate sorption distribution constant, Kd. Extractable Cd concentrations in soil had no significant effect on glyphosate sorption. Glyphosate Kd values significantly decreased with increasing Olsen-P concentrations in soil, regardless of the pH conditions studied. Experiments repeated with a commercially available glyphosate formulation showed statistically similar results as the experiments performed with analytical-grade glyphosate. Co-applications of MAP with glyphosate also reduced the available sorption sites to retain glyphosate, but less so when soils already contain large amounts of phosphate. Glyphosate Kd values in soils ranged from 173 to 939 L kg(-1) under very strong to strongly acidic condition but the Kd was always <100 L kg(-1) under moderately acidic to slightly alkaline conditions. The highest Olsen-P concentrations in soil reduced Kd values by 25-44% relative to control soils suggesting that, under moderately acidic to slightly alkaline conditions, glyphosate may become mobile by water in soils with high phosphate levels. Otherwise, glyphosate residues in agricultural soils are more likely to be transported off-site by wind and water-eroded sediments than by leaching or runoff. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Online assay of bone specific alkaline phosphatase with a flow injection-bead injection system.

    PubMed

    Hartwell, Supaporn Kradtap; Somprayoon, Duangporn; Kongtawelert, Prachya; Ongchai, Siriwan; Arppornchayanon, Olarn; Ganranoo, Lucksagoon; Lapanantnoppakhun, Somchai; Grudpan, Kate

    2007-09-26

    Alkaline phosphatase (ALP) has been used as one of the biomarkers for bone resorption and liver diseases. Normally, total alkaline phosphatase is quantified along with other symptoms to determine the releasing source of the alkaline phosphatase. A semi-automated flow injection-bead injection system was proposed to conveniently and selectively assay bone alkaline phosphatase (BALP) based on its specific binding to wheat germ coated beads. Amount of BALP in serum was determined from the intensity of the yellow product produced from bound BALP on the retained beads and its substrate pNPP. The used beads were discarded and the fresh ones were introduced for the next analysis. The reaction cell was designed to be opened and closed using a computer controlled solenoid valve for a precise incubation time. The performance of the proposed system was evaluated by using it to assay BALP in human serum. The results were compared to those obtained by using a commercial ELISA kit. The system is proposed to be an easy and cost effective system for quantification of BALP as an alternative to batch wise wheat germ specific binding technique.

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

    PubMed

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

    2016-12-01

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

  12. Gingival crevicular fluid alkaline phosphatase activity reflects periodontal healing/recurrent inflammation phases in chronic periodontitis patients.

    PubMed

    Perinetti, Giuseppe; Paolantonio, Michele; Femminella, Beatrice; Serra, Emanuela; Spoto, Giuseppe

    2008-07-01

    Roles for host enzymes as diagnostic indicators of periodontal status in gingival crevicular fluid (GCF) have been proposed. One of these host enzymes is alkaline phosphatase (ALP), the GCF activity of which has been associated with periodontal inflammation. Thus, the present study aimed to improve our understanding of how the healing of chronic periodontitis following scaling and root planing (SRP) affects GCF ALP activity after 15 and 60 days. Sixteen systemically healthy subjects (aged 35 to 61 years) with moderate to advanced generalized chronic periodontitis were recruited. In each subject, paired pockets with probing depths (PDs) > or =4 mm that were located in two symmetric quadrants were chosen. These sites were randomized at the split-mouth level, with half receiving SRP treatment and the other half left untreated. Ninety-two pockets were included in the study. Clinical examinations were performed at baseline (prior to SRP) and after 15 and 60 days; information recorded included the presence of plaque, PD, clinical attachment level (CAL), and bleeding on probing. GCF was collected from each pocket included in the study at the three time points. A large and significant decrease in GCF ALP activity was seen 15 days after SRP, concomitant with an improvement in clinical parameters. After 60 days, an increase in GCF ALP activity back to baseline levels was recorded along with further improvements in clinical parameters. Moreover, in the SRP pockets with initial PDs >6 mm, the CAL gains between days 15 and 60 were significantly associated with changes in GCF ALP activity over the same time interval. The decrease in GCF ALP activity at 15 days corresponded to a decrease in clinical signs of inflammation; in contrast, the increase in GCF ALP activity at 60 days seemed to be related to subclinical recurrent inflammation or further healing/remodeling of the periodontal tissue. Therefore, GCF ALP reflects the short-term periodontal healing/recurrent inflammation

  13. Evaluation of non-specific binding suppression schemes for neutravidin and alkaline phosphatase at the surface of reticulated vitreous carbon electrodes.

    PubMed

    Shedge, Hemangi Y; Creager, Stephen E

    2010-01-11

    Non-specific binding (NSB) of high-molecular-weight proteins onto electrode surfaces can complicate the application of electroanalytical techniques to clinical and environmental research, particularly in biosensor applications. We present herein various strategies to modify the surface of reticulated vitreous carbon (RVC) electrodes to suppress non-specific binding of biomolecules onto its surface. Non-specific binding and specific binding (SB) of two enzyme conjugates, neutravidin-alkaline phosphatase (NA-ALP) and biotinylated alkaline phosphatase (B-ALP), and also neutravidin itself, were studied using hydroquinone diphosphate (HQDP) as an enzyme substrate for ALP inside the pores of RVC electrodes that had been subjected to various modification schemes. The extent of NSB and SB of these biomolecules inside RVC pores was assessed by measuring the initial rate of generation of an electroactive product, hydroquinone (HQ), of the enzyme-catalyzed reaction, using linear scan voltammetry (LSV) for HQ detection. Electrodes functionalized with phenylacetic acid and poly(ethylene glycol) (PEG) showed low NSB and high SB (when biotin capture ligands were included in the modification scheme) in comparison with unmodified electrodes and RVC electrodes modified in other ways. A simple sandwich bioassay for neutravidin was performed on the RVC electrode with the lowest NSB. A concentration detection limit of 52+/-2 ng mL(-1) and an absolute detection limit of 5.2+/-0.2 ng were achieved for neutravidin when this assay was performed using a 100 microL sample size.

  14. Isolation and characterization of two immunochemically distinct alkaline phosphatases from Pseudomonas aeruginosa.

    PubMed

    Tan, A S; Worobec, E A

    1993-02-01

    We have isolated two alkaline phosphatases (H-AP and L-AP, for high and low molecular mass, respectively) from Pseudomonas aeruginosa PA01. These two enzymes were found to differ in mobility on sodium dodecyl sulphate polyacrylamide gels (H-AP, M(r) = 51,000 and L-AP, M(r) = 39,500), amino-terminal amino acid sequence and did not cross-react. Both enzymes were active as phosphomonoesterases while only L-AP demonstrated any phosphodiesterase activity. Both enzymes were purified from P. aeruginosa grown in phosphate limiting conditions using the same protocol and were identified in both periplasmic and extracellular locations. A low level of H-AP was produced constitutively whereas L-AP was produced only after induction by reduced phosphate concentration in the growth medium. An L-AP-like enzyme has been previously described, however, this is the first report of a second P. aeruginosa alkaline phosphatase.

  15. Effects of tunicamycin, mannosamine, and other inhibitors of glycoprotein processing on skeletal alkaline phosphatase in human osteoblast-like cells.

    PubMed

    Farley, J R; Magnusson, P

    2005-01-01

    Skeletal alkaline phosphatase (sALP) is a glycoprotein- approximately 20% carbohydrate by weight, with five presumptive sites for N-linked glycosylation, as well as a carboxy-terminal site for attachment of the glycolipid structure (glycosylphosphatidylinositol, GPI), which anchors sALP to the outer surface of osteoblasts. The current studies were intended to characterize the effects of inhibiting glycosylation and glycosyl-processing on the synthesis, plasma membrane attachment, cellular-extracellular distribution, and reaction kinetics of sALP in human osteosarcoma (SaOS-2) cells. sALP synthesis, glycosylation, and GPI-anchor attachment were assessed as total protein synthesis/immunospecific sALP synthesis, sialic acid content (i.e., wheat germ agglutinin precipitation), and insolubility (i.e., temperature-dependent phase-separation), respectively. sALP reaction kinetics were characterized by analysis of dose-dependent initial velocity data, with a phosphoryl substrate. The results of these studies revealed that the inhibition of either N-linked glycosylation or oligosaccharide synthesis for GPI-anchor addition could affect the synthesis and the distribution of sALP, but not the kinetics of the phosphatase reaction. Tunicamycin-which blocks N-linked glycosylation by inhibiting core oligosaccharide synthesis-decreased cell layer protein and the total amount of sALP in the cells, while increasing the relative level of sALP in the cell-conditioned culture medium (CM, i.e., the amount of sALP released). These effects were attributed to dose- and time-dependent decreases in sALP synthesis and N-linked glycosylation, and an increase in apoptotic cell death (P <0.001 for each). In contrast to the effects of tunicamycin on N-linked glycosylation, the effects of mannosamine, which inhibits GPI-anchor glycosylation/formation, included (1) an increase in cell layer protein; (2) decreases in sALP specific activity, in the cells and in the CM; and (3) increases in the

  16. Serum alkaline phosphatase as a predictor of worsening renal function in patients with acute decompensated heart failure.

    PubMed

    Yamazoe, Masahiro; Mizuno, Atsushi; Nishi, Yutaro; Niwa, Koichiro; Isobe, Mitsuaki

    2016-05-01

    Venous congestion has come into focus as an important hemodynamic factor for worsening renal function (WRF) in patients with acute decompensated heart failure (ADHF). Serum alkaline phosphatase (ALP) was reported as a biological marker of liver congestion in ADHF. The purpose of this study was to determine whether ALP is a predictor of WRF in patients with ADHF. We enrolled consecutive patients admitted to a single cardiovascular center with ADHF, and defined WRF as an increase in creatinine of >0.3 mg/dl during hospitalization and chronic kidney disease (CKD) as an estimated glomerular filtration rate (eGFR) <60 ml/min/1.73 m(2). The patients were classified into tertiles by ALP level (<203, 203-278, and >278 IU/L). A total of 972 patients (mean age, 76±13 years; 54% male) were retrospectively analyzed. WRF was identified in 132 patients (13.6%). In multivariate logistic regression analysis, baseline CKD [odds ratio (OR) 2.46, 95% confidence interval (CI) 1.48-4.08, p<0.001], serum albumin (OR 0.52, 95% CI 0.35-0.77, p=0.001), and diabetes (OR 2.07, 95% CI 1.37-3.12, p<0.001) were associated with WRF. Compared with the lowest tertile (ALP <203 IU/L), an adjusted OR of WRF was 1.69 (95% CI 1.02-2.79, p=0.04) in the middle tertile (ALP, 203-278 IU/L) and 1.95 (95% CI 1.20-3.21, p=0.008) in the highest tertile (ALP >278 IU/L). Serum ALP is an independent predictor of WRF in the clinical course of ADHF. Copyright © 2015 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sakisaka, Yukihiko; Tsuchiya, Masahiro; Tohoku Fukushi University, Sendai 989-3201

    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 genemore » 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.« less

  18. The ALP miracle: unified inflaton and dark matter

    NASA Astrophysics Data System (ADS)

    Daido, Ryuji; Takahashi, Fuminobu; Yin, Wen

    2017-05-01

    We propose a scenario where both inflation and dark matter are described by a single axion-like particle (ALP) in a unified manner. In a class of the minimal axion hilltop inflation, the effective masses at the maximum and mimimum of the potential have equal magnitude but opposite sign, so that the ALP inflaton is light both during inflation and in the true vacuum. After inflation, most of the ALPs decay and evaporate into plasma through a coupling to photons, and the remaining ones become dark matter. We find that the observed CMB and matter power spectrum as well as the dark matter abundance point to an ALP of mass mphi = Script O(0.01) eV and the axion-photon coupling gphi γ γ = Script O(10-11) GeV-1: the ALP miracle. The suggested parameter region is within the reach of the next generation axion helioscope, IAXO, and high-intensity laser experiments in the future. Furthermore, thermalized ALPs contribute to hot dark matter and its abundance is given in terms of the effective number of extra neutrino species, Δ Neff simeq 0.03, which can be tested by the future CMB and BAO observations. We also discuss a case with multiple ALPs, where the coupling to photons can be enhanced in the early Universe by an order of magnitude or more, which enlarges the parameter space for the ALP miracle. The heavy ALP plays a role of the waterfall field in hybrid inflation, and reheats the Universe, and it can be searched for in various experiments such as SHiP.

  19. Enrichment of enzymatically mineralized gellan gum hydrogels with phlorotannin-rich Ecklonia cava extract Seanol(®) to endow antibacterial properties and promote mineralization.

    PubMed

    Douglas, Timothy E L; Dokupil, Agnieszka; Reczyńska, Katarzyna; Brackman, Gilles; Krok-Borkowicz, Malgorzata; Keppler, Julia K; Božič, Mojca; Van Der Voort, Pascal; Pietryga, Krzysztof; Samal, Sangram Keshari; Balcaen, Lieve; van den Bulcke, Jan; Van Acker, Joris; Vanhaecke, Frank; Schwarz, Karin; Coenye, Tom; Pamuła, Elżbieta

    2016-08-10

    Hydrogels offer several advantages as biomaterials for bone regeneration, including ease of incorporation of soluble substances such as mineralization-promoting enzymes and antibacterial agents. Mineralization with calcium phosphate (CaP) increases bioactivity, while antibacterial activity reduces the risk of infection. Here, gellan gum (GG) hydrogels were enriched with alkaline phosphatase (ALP) and/or Seanol(®), a seaweed extract rich in phlorotannins (brown algae-derived polyphenols), to induce mineralization with CaP and increase antibacterial activity, respectively. The sample groups were unmineralized hydrogels, denoted as GG, GG/ALP, GG/Seanol and GG/Seanol/ALP, and hydrogels incubated in mineralization medium (0.1 M calcium glycerophosphate), denoted as GG/ALP_min, GG/Seanol_min and GG/Seanol/ALP_min. Seanol(®) enhanced mineralization with CaP and also increased compressive modulus. Seanol(®) and ALP interacted in a non-covalent manner. Release of Seanol(®) occurred in a burst phase and was impeded by ALP-mediated mineralization. Groups GG/Seanol and GG/ALP/Seanol exhibited antibacterial activity against methicillin-resistant Staphylococcus aureus. GG/Seanol/ALP_min, but not GG/Seanol_min, retained some antibacterial activity. Eluates taken from groups GG/ALP_min, GG/Seanol_min and GG/ALP/Seanol_min displayed comparable cytotoxicity towards MG-63 osteoblast-like cells. These results suggest that enrichment of hydrogel biomaterials with phlorotannin-rich extracts is a promising strategy to increase mineralizability and antibacterial activity.

  20. Osteoblast Differentiation on Collagen Scaffold with Immobilized Alkaline Phosphatase.

    PubMed

    Jafary, F; Hanachi, P; Gorjipour, K

    2017-01-01

    In tissue engineering, scaffold characteristics play an important role in the biological interactions between cells and the scaffold. Cell adhesion, proliferation, and activation depend on material properties used for the fabrication of scaffolds. In the present investigation, we used collagen with proper characteristics including mechanically stability, biodegradability and low antigenicity. Optimization of the scaffold was done by immobilization of alkaline phosphatase on the collagen surface via cross-linking method, because this enzyme is one of the most important markers of osteoblast, which increases inorganic phosphate concentration and promote mineralization of bone formation. Alkaline phosphatase was immobilized on a collagen surface by 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride, as a reagent. Then, rat mesenchymal stem cells were cultured in osteogenic medium in control and treated groups. The osteogenesis-related genes were compared between treatments (differentiated cells with immobilized alkaline phosphatase/collagen scaffold) and control groups (differentiated cells on collagen surface without alkaline phosphatase) on days 3 and 7 by quantitative real-time PCR (QIAGEN software). Several genes, including alkaline phosphatase, collagen type I and osteocalcine associated with calcium binding and mineralization, showed upregulation in expression during the first 3 days, whereas tumor necrosis factor-α, acting as an inhibitor of differentiation, was down-regulated during osteogenesis. Collagen scaffold with immobilized alkaline phosphatase can be utilized as a good candidate for enhancing the differentiation of osteoblasts from mesenchymal stem cells.

  1. Age and prematurity of the Alps

    NASA Astrophysics Data System (ADS)

    Hergarten, Stefan; Stüwe, Kurt; Wagner, Thomas

    2010-05-01

    Although the Alps are among the best studied mountain ranges on Earth, the age of their topography is almost unknown. Even their relative stage of evolution is unclear: Are the Alps still growing, in a steady state or even decaying? Using the mean slope at given catchment size as a new geomorphic parameter we analyse the topography of the Alps. Our analysis provides one of the first quantitative constraints that shows that the range is still in its infancy: In contrast to several other mountain ranges, the Alps have still more than half of their evolution to a geomorphic steady state to go. Combining our results with sediment data from the surrounding accumulation spaces we infer that the formation of substantial topography began only 5-6 million years ago. Our results challenge a general consensus that the topographic evolution is distributed over much of the Miocene.

  2. Injectable TEMPO-oxidized nanofibrillated cellulose/biphasic calcium phosphate hydrogel for bone regeneration.

    PubMed

    Safwat, Engie; Hassan, Mohammad L; Saniour, Sayed; Zaki, Dalia Yehia; Eldeftar, Mervat; Saba, Dalia; Zazou, Mohamed

    2018-05-01

    Nanofibrillated cellulose, obtained from rice straw agricultural wastes was used as a substrate for the preparation of a new injectable and mineralized hydrogel for bone regeneration. Tetramethyl pyridine oxyl (TEMPO) oxidized nanofibrillated cellulose, was mineralized through the incorporation of a prepared and characterized biphasic calcium phosphate at a fixed ratio of 50 wt%. The TEMPO-oxidized rice straw nanofibrillated cellulose was characterized using transmission electron microscopy, Fourier transform infrared, and carboxylic content determination. The injectability and viscosity of the prepared hydrogel were evaluated using universal testing machine and rheometer testing, respectively. Cytotoxicity and alkaline phosphatase level tests on osteoblast like-cells for in vitro assessment of the biocompatibility were investigated. Results revealed that the isolated rice straw nanofibrillated cellulose is a nanocomposite of the cellulose nanofibers and silica nanoparticles. Rheological properties of the tested materials are suitable for use as injectable material and of nontoxic effect on osteoblast-like cells, as revealed by the positive alkaline phosphate assay. However, nanofibrillated cellulose/ biphasic calcium phosphate hydrogel showed higher cytotoxicity and lower bioactivity test results when compared to that of nanofibrillated cellulose.

  3. Ivrea mantle wedge and arc of the Western Alps (II): Kinematic evolution of the Alps-Apennines orogenic system

    NASA Astrophysics Data System (ADS)

    Schmid, Stefan; Kissling, Eduard; van Hinsbergen, Douwe J. J.; Molli, Giancarlo

    2017-04-01

    Integration of geological and geophysical data on the deep structure of the Alps (Kissling et al. 2017) reveals that the deep-seated Ivrea mantle played a crucial role during the formation of the arc of the Western Alps. Exhumation of the mantle beneath the Ivrea Zone to shallow crustal depths during Mesozoic rifting led to the formation of a strong Ivrea mantle wedge; its strength exceeds that of surrounding mostly quartz-bearing units, and consequently allows for indentation and wedging of a quasi-rigid Ivrea mantle wedge into the Western Alps during Alpine orogeny. A first early stage (pre-35 Ma) of the West-Alpine orogenic evolution is characterized by top-NNW thrusting in sinistral transpression causing at least some 260km displacement of internal Western Alps and E-W-striking Alps farther east, together with the Adria micro-plate, towards N to NNW with respect to stable Europe. It is during the second stage (35-25 Ma) that the Ivrea mantle wedge played a crucial role by accentuating the arc. This stage is associated with top-WNW thrusting in the external zones of the central portion of the arc and lateral indentation and wedging of the Ivrea mantle slice beneath the already existing nappe pile towards WNW by some 100-150km. The final stage of arc formation (25-0 Ma) is associated with orogeny in the Apennines leading to oroclinal bending in the southernmost Western Alps that by now became parts of the Apenninic orogen, in connection with the 50° counterclockwise rotation of the Corsica-Sardinia block and the Ligurian Alps. The lithological composition of some tectonic units originating from the Alpine Tethys (Piemont-Liguria Ocean) found in the Alps and the northern Apennines has much in common. The non-metamorphic parts of the Piemont-Liguria derived units form the upper plate of the Western Alps that is devoid of Austroalpine elements, while the lower plate includes highly metamorphic units derived from the same Piemont-Liguria Ocean. This points to a

  4. Gingival crevicular fluid alkaline phosphatase activity as a non-invasive biomarker of skeletal maturation.

    PubMed

    Perinetti, G; Baccetti, T; Contardo, L; Di Lenarda, R

    2011-02-01

    To evaluate the gingival crevicular fluid (GCF) alkaline phosphatase (ALP) activity in growing subjects in relation to the stages of individual skeletal maturation. The Department of Biomedicine, University of Trieste. Seventy-two healthy growing subjects (45 women and 27 men; range, 7.8-17.7 years). Double-blind, prospective, cross-sectional design. Samples of GCF were collected from each subject at the mesial and distal sites of both of the central incisors, in the maxilla and mandible. Skeletal maturation phase was assessed through the cervical vertebral maturation (CVM) method. Enzymatic activity was determined spectrophotometrically. The relationship between GCF ALP activity and CVM stages was significant. In particular, a twofold peak in enzyme activity was seen at the CS3 and CS4 pubertal stages, compared to the pre-pubertal stages (CS1 and CS2) and post-pubertal stages (CS5 and CS6), at both the maxillary and mandibular sites. No differences were seen between the maxillary and mandibular sites, or between the sexes. As an adjunct to standard methods based upon radiographic parameters, the GCF ALP may be a candidate as a non-invasive clinical biomarker for the identification of the pubertal growth spurt in periodontally healthy subjects scheduled for orthodontic treatment. © 2010 John Wiley & Sons A/S.

  5. Studies on the interactions between purified bovine caseins and alkaline-earth-metalions

    PubMed Central

    Dickson, I. R.; Perkins, D. J.

    1971-01-01

    1. Alkaline-earth-metal cations at low concentrations form soluble complexes with bovine caseins. The relative order of binding capacities is: Mg2+>Ca2+>Ba2+>Sr2+. 2. The cations interact with both free ionized carboxyl groups of aspartic acid and glutamic acid and with monoester phosphate groups covalently bound to serine and threonine; at low concentrations of the cations interactions are predominantly with the phosphate groups. 3. The order of binding capacities for purified components of the casein complex is: αs1-casein>β-casein>κ-casein. PMID:5166590

  6. A Development Testbed for ALPS-Based Systems

    DTIC Science & Technology

    1988-10-01

    alloted to tile application because of size or power constraints). Given an underlying support ALPS architecture such as the d-ALPS architecture, a...resource on which it is assigned at runtime. A second representation problem is that most graph analysis algorithms treat either graphs with weighted links...subtask) associated with it but is treated like other links. In d-ALPS, as a priority precedence link, it would cause the binding of a pro- cessor: as a

  7. Osteoinduction and survival of osteoblasts and bone-marrow stromal cells in 3D biphasic calcium phosphate scaffolds under static and dynamic culture conditions.

    PubMed

    Rath, Subha N; Strobel, Leonie A; Arkudas, Andreas; Beier, Justus P; Maier, Anne-Kathrin; Greil, Peter; Horch, Raymund E; Kneser, Ulrich

    2012-10-01

    In many tissue engineering approaches, the basic difference between in vitro and in vivo conditions for cells within three-dimensional (3D) constructs is the nutrition flow dynamics. To achieve comparable results in vitro, bioreactors are advised for improved cell survival, as they are able to provide a controlled flow through the scaffold. We hypothesize that a bioreactor would enhance long-term differentiation conditions of osteogenic cells in 3D scaffolds. To achieve this either primary rat osteoblasts or bone marrow stromal cells (BMSC) were implanted on uniform-sized biphasic calcium phosphate (BCP) scaffolds produced by a 3D printing method. Three types of culture conditions were applied: static culture without osteoinduction (Group A); static culture with osteoinduction (Group B); dynamic culture with osteoinduction (Group C). After 3 and 6 weeks, the scaffolds were analysed by alkaline phosphatase (ALP), dsDNA amount, SEM, fluorescent labelled live-dead assay, and real-time RT-PCR in addition to weekly alamarBlue assays. With osteoinduction, increased ALP values and calcium deposition are observed; however, under static conditions, a significant decrease in the cell number on the biomaterial is observed. Interestingly, the bioreactor system not only reversed the decreased cell numbers but also increased their differentiation potential. We conclude from this study that a continuous flow bioreactor not only preserves the number of osteogenic cells but also keeps their differentiation ability in balance providing a suitable cell-seeded scaffold product for applications in regenerative medicine. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.

  8. Phosphate Solubilization Potential and Phosphatase Activity Of Rhizospheric Trichoderma Spp.

    PubMed Central

    Anil, Kapri; Lakshmi, Tewari

    2010-01-01

    Trichoderma sp., a well known biological control agent against several phytopathogens, was tested for its phosphate (P) solubilizing potential. Fourteen strains of Trichoderma sp. were isolated from the forest tree rhizospheres of pinus, deodar, bamboo, guava and oak on Trichoderma selective medium. The isolates were tested for their in-vitro P-solubilizing potential using National Botanical Research Institute Phosphate (NBRIP) broth containing tricalcium phosphate (TCP) as the sole P source, and compared with a standard culture of T. harzianum. All the cultures were found to solubilize TCP but with varying potential. The isolate DRT-1 showed maximum amount of soluble phosphate (404.07 εg.ml-1), followed by the standard culture of T. harzianum (386.42 εg.ml-1) after 96 h of incubation at 30±10C. Extra-cellular acid and alkaline phosphatases of the fungus were induced only in the presence of insoluble phosphorus source (TCP). High extra-cellular alkaline phosphatase activity was recorded for the isolate DRT-1 (14.50 U.ml-1) followed by the standard culture (13.41 U.ml-1) at 72h. The cultures showed much lesser acid phosphatase activities. Under glasshouse conditions, Trichoderma sp. inoculation increased chickpea (Cicer arietinum) growth parameters including shoot length, root length, fresh and dry weight of shoot as well as roots, in P-deficient soil containing only bound phosphate (TCP). Shoot weight was increased by 23% and 33% by inoculation with the isolate DRT-1 in the soil amended with 100 and 200 mg TCP kg-1 soil, respectively, after 60 d of sowing. The study explores high P-solubilizing potential of Trichoderma sp., which can be exploited for the solubilization of fixed phosphates present in the soil, thereby enhancing soil fertility and plant growth. PMID:24031556

  9. Phosphatase activity in Antarctica soil samples as a biosignature of extant life

    NASA Astrophysics Data System (ADS)

    Sato, Shuji; Itoh, Yuki; Takano, Yoshinori; Fukui, Manabu; Kaneko, Takeo; Kobayashi, Kensei

    Microbial activities have been detected in such extreme terrestrial environments as deep lithosphere, a submarine hydrothermal systems, stratosphere, and Antarctica. Microorganisms have adapted to such harsh environments by evolving their biomolecules. Some of these biomolecules such as enzymes might have different characteristics from those of organisms in ordinary environments. Many biosignatures (or biomarkers) have been proposed to detect microbial activities in such extreme environments. A number of techniques are proposed to evaluate biological activities in extreme environments including cultivation methods, assay of metabolism, and analysis of bioorganic compounds like amino acids and DNA. Enzyme activities are useful signature of extant life in extreme environments. Among many enzymes, phosphatase could be a good indicator of biological activities, since phosphate esters are essential for all the living terrestrial organisms. In addition, alkaline phosphatase is known as a typical zinc-containing metalloenzyme and quite stable in environments. We analyzed phosphatase activities in Antarctica soil samples to see whether they can be used as biosignatures for extant life. In addition, we characterized phosphatases extracted from the Antarctica soil samples, and compared with those obtained from other types of environments. Antarctica surface environments are quite severe environments for life since it is extremely cold and dry and exposed to strong UV and cosmic rays. We tried to evaluate biological activities in Antarctica by measuring phosphatase activities. Surface soil samples are obtained at the Sites 1-8 near Showa Base in Antarctica during the 47th Japan Antarctic exploration mission in 2005-6. Activities of acid phosphatase (ACP) and alkaline phosphatase (ALP) are measured spectrophotometrically after mixing the powdered sample and p-nitrophenyl phosphate solution (pH 6.5 for ACP, pH 8.0 for ALP). ALP was characterized after extraction from soils with

  10. Degradation of Chlorpyrifos by an alkaline phosphatase from the cyanobacterium Spirulina platensis.

    PubMed

    Thengodkar, Rutwik Ravindra Mandakini; Sivakami, S

    2010-07-01

    Spirulina is a photosynthetic, filamentous, spiral-shaped, multicellular, blue-green microalga. The two most important species are Spirulina maxima and Spirulina platensis. Spirulina is considered an excellent food, lacking toxicity and having corrective properties against viral attacks, anemia, tumor growth and malnutrition. We have observed that cultures of Spirulina platensis grow in media containing up to 80 ppm of the organophosphorous pesticide, Chlorpyrifos. It was found to be due to an alkaline phosphatase (ALP) activity that was detected in cell free extracts of Spirulina platensis. This activity was purified from the cell free extracts using ammonium sulphate precipitation and gel filtration and shown to belong to the class of EC 3.1.3.1 ALP. The purified enzyme degrades 100 ppm Chlorpyrifos to 20 ppm in 1 h transforming it into its primary metabolite 3, 5, 6-trichloro-2-pyridinol. This is the first report of degradation of Chlorpyrifos by Spirulina platensis whose enzymic mechanism has been clearly identified. These findings have immense potential for harnessing Spirulina platensis in bioremediation of polluted ecosystems.

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

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

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

  12. Tricalcium phosphate and glutaraldehyde crosslinked gelatin incorporating bone morphogenetic protein--a viable scaffold for bone tissue engineering.

    PubMed

    Yang, Shu-Hua; Hsu, Chung-King; Wang, Kuo-Cheng; Hou, Sheng-Mou; Lin, Feng-Huei

    2005-07-01

    Bone defects caused by various etiologies must be filled with suitable substances to promote bone repair. Autogenous iliac crest graft is most frequently used, but is often associated with morbidities. Several bone graft substitutes have been developed to provide osteoconductive matrices as well as to enhance osteoinductivity. A tricalcium phosphate and glutaraldehyde crosslinked gelatin (GTG) scaffold, incorporated with bone morphogenetic proteins (BMPs), was developed to provide an alternative mean of bone tissue engineering. This study investigated differences between GTG and BMP-4 immobilized GTG (GTG-BMP) scaffolds on neonatal rat calvaria osteoblast activities. The GTG scaffold possessed an average pore size of 200 microm and a porosity of 75%. HE staining revealed uniform cell distribution throughout the scaffold 24 h post cell seeding. Alkaline phosphatase (ALP) activity of the GTG samples increased initially and then stabilized at 3 weeks postseeding. ALP activity of the GTG-BMP samples was similar to that of the GTG samples in the second and third weeks, but it continued increasing and became significantly greater than that of the GTG samples by the fourth week. Gla-type osteocalcin (Gla-OC) activity of the GTG-BMP samples was initially lower, but also became significantly greater than that of the GTG samples by the fourth week. An HE stain revealed greater numbers of attached cells and a richer matrix deposits in the GTG-BMP samples. A von Kossa stain showed larger mineralizing nodules, in greater numbers, after 4 weeks of in vitro cultivation. These findings suggest that the GTG scaffold provides an excellent porous structure, conductive to greater cell attachment and osteoblast differentiation, and that utility can be significantly enhanced by the inclusion of BMPs. A GTG-BMP scaffold holds promise as a superior bioactive material for bone tissue engineering. Copyright 2005 Wiley Periodicals, Inc.

  13. Ovarian cancer stem-like cells with induced translineage-differentiation capacity and are suppressed by alkaline phosphatase inhibitor

    PubMed Central

    Liu, Kuei-Chun; Yo, Yi-Te; Huang, Rui-Lan; Wang, Yu-Chi; Liao, Yu-Ping; Huang, Tien-Shuo; Chao, Tai-Kuang; Lin, Chi-Kang; Weng, Shao-Ju; Ma, Kuo-Hsing; Chang, Cheng-Chang; Yu, Mu-Hsien; Lai, Hung-Cheng

    2013-01-01

    Spheroid formation is one property of stem cells—such as embryo-derived or neural stem cells—that has been used for the enrichment of cancer stem-like cells (CSLCs). However, it is unclear whether CSLC-derived spheroids are heterogeneous or whether they share common embryonic stemness properties. Understanding these features might lead to novel therapeutic approaches. Ovarian carcinoma is a deadly disease of women. We identified two types of spheroids (SR1 and SR2) from ovarian cancer cell lines and patients' specimens according to their morphology. Both types expressed stemness markers and could self-renew and initiate tumors when a low number of cells were used. Only SR1 could differentiate into multiple-lineage cell types under specific induction conditions. SR1 spheroids could differentiate to SR2 spheroids through epithelial–mesenchymal transition. Alkaline phosphatase (ALP) was highly expressed in SR1 spheroids, decreased in SR2 spheroids, and was absent in differentiated progenies in accordance with the loss of stemness properties. We verified that ALP can be a marker for ovarian CSLCs, and patients with greater ALP expression is related to advanced clinical stages and have a higher risk of recurrence and lower survival rate. The ALP inhibitor, levamisole, disrupted the self-renewal of ovarian CSLCs in vitro and tumor growth in vivo. In summary, this research provides a plastic ovarian cancer stem cell model and a new understanding of the cross-link between stem cells and cancers. This results show that ovarian CSLCs can be suppressed by levamisole. Our findings demonstrated that some ovarian CSLCs may restore ALP activity, and this suggests that inhibition of ALP activity may present a new opportunity for treatment of ovarian cancer. PMID:24280306

  14. Alendronate-Eluting Biphasic Calcium Phosphate (BCP) Scaffolds Stimulate Osteogenic Differentiation

    PubMed Central

    Kim, Sung Eun; Lee, Deok-Won; Kang, Eun Young; Jeong, Won Jae; Lee, Boram; Jeong, Myeong Seon; Kim, Hak Jun; Park, Kyeongsoon; Song, Hae-Ryong

    2015-01-01

    Biphasic calcium phosphate (BCP) scaffolds have been widely used in orthopedic and dental fields as osteoconductive bone substitutes. However, BCP scaffolds are not satisfactory for the stimulation of osteogenic differentiation and maturation. To enhance osteogenic differentiation, we prepared alendronate- (ALN-) eluting BCP scaffolds. The coating of ALN on BCP scaffolds was confirmed by scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDS), and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). An in vitro release study showed that release of ALN from ALN-eluting BCP scaffolds was sustained for up to 28 days. In vitro results revealed that MG-63 cells grown on ALN-eluting BCP scaffolds exhibited increased ALP activity and calcium deposition and upregulated gene expression of Runx2, ALP, OCN, and OPN compared with the BCP scaffold alone. Therefore, this study suggests that ALN-eluting BCP scaffolds have the potential to effectively stimulate osteogenic differentiation. PMID:26221587

  15. Influence of triethyl phosphate on phosphatase activity in shooting range soil: Isolation of a zinc-resistant bacterium with an acid phosphatase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Story, Sandra; Brigmon, Robin L.

    Phosphatase-mediated hydrolysis of organic phosphate may be a viable means of stabilizing heavy metals via precipitation as a metal phosphate in bioremediation applications. We investigated the effect of triethyl phosphate (TEP) on soil microbial-phosphatase activity in a heavy-metal contaminated soil. Gaseous TEP has been used at subsurface sites for bioremediation of organic contaminants but not applied in heavy-metal contaminated areas. Little is known about how TEP affects microbial activity in soils and it is postulated that TEP can serve as a phosphate source in nutrient-poor groundwater and soil/sediments. Over a 3-week period, TEP amendment to microcosms containing heavy-metal contaminated soilmore » resulted in increased activity of soil acid-phosphatase and repression of alkaline phosphatase, indicating a stimulatory effect on the microbial population. A soil-free enrichment of microorganisms adapted to heavy-metal and acidic conditions was derived from the TEP-amended soil microcosms using TEP as the sole phosphate source and the selected microbial consortium maintained a high acid-phosphatase activity with repression of alkaline phosphatase. Addition of 5 mM zinc to soil-free microcosms had little effect on acid phosphatase but inhibited alkaline phosphatase. One bacterial member from the consortium, identified as Burkholderia cepacia sp., expressed an acid-phosphatase activity uninhibited by high concentrations of zinc and produced a soluble, indigo pigment under phosphate limitation. The pigment was produced in a phosphate-free medium and was not produced in the presence of TEP or phosphate ion, indicative of purple acid-phosphatase types that are pressed by bioavailable phosphate. Finally, these results demonstrate that TEP amendment was bioavailable and increased overall phosphatase activity in both soil and soil-free microcosms supporting the possibility of positive outcomes in bioremediation applications.« less

  16. Influence of triethyl phosphate on phosphatase activity in shooting range soil: Isolation of a zinc-resistant bacterium with an acid phosphatase

    DOE PAGES

    Story, Sandra; Brigmon, Robin L.

    2016-12-19

    Phosphatase-mediated hydrolysis of organic phosphate may be a viable means of stabilizing heavy metals via precipitation as a metal phosphate in bioremediation applications. We investigated the effect of triethyl phosphate (TEP) on soil microbial-phosphatase activity in a heavy-metal contaminated soil. Gaseous TEP has been used at subsurface sites for bioremediation of organic contaminants but not applied in heavy-metal contaminated areas. Little is known about how TEP affects microbial activity in soils and it is postulated that TEP can serve as a phosphate source in nutrient-poor groundwater and soil/sediments. Over a 3-week period, TEP amendment to microcosms containing heavy-metal contaminated soilmore » resulted in increased activity of soil acid-phosphatase and repression of alkaline phosphatase, indicating a stimulatory effect on the microbial population. A soil-free enrichment of microorganisms adapted to heavy-metal and acidic conditions was derived from the TEP-amended soil microcosms using TEP as the sole phosphate source and the selected microbial consortium maintained a high acid-phosphatase activity with repression of alkaline phosphatase. Addition of 5 mM zinc to soil-free microcosms had little effect on acid phosphatase but inhibited alkaline phosphatase. One bacterial member from the consortium, identified as Burkholderia cepacia sp., expressed an acid-phosphatase activity uninhibited by high concentrations of zinc and produced a soluble, indigo pigment under phosphate limitation. The pigment was produced in a phosphate-free medium and was not produced in the presence of TEP or phosphate ion, indicative of purple acid-phosphatase types that are pressed by bioavailable phosphate. Finally, these results demonstrate that TEP amendment was bioavailable and increased overall phosphatase activity in both soil and soil-free microcosms supporting the possibility of positive outcomes in bioremediation applications.« less

  17. Influence of triethyl phosphate on phosphatase activity in shooting range soil: Isolation of a zinc-resistant bacterium with an acid phosphatase.

    PubMed

    Story, Sandra; Brigmon, Robin L

    2017-03-01

    Phosphatase-mediated hydrolysis of organic phosphate may be a viable means of stabilizing heavy metals via precipitation as a metal phosphate in bioremediation applications. We investigated the effect of triethyl phosphate (TEP) on soil microbial-phosphatase activity in a heavy-metal contaminated soil. Gaseous TEP has been used at subsurface sites for bioremediation of organic contaminants but not applied in heavy-metal contaminated areas. Little is known about how TEP affects microbial activity in soils and it is postulated that TEP can serve as a phosphate source in nutrient-poor groundwater and soil/sediments. Over a 3-week period, TEP amendment to microcosms containing heavy-metal contaminated soil resulted in increased activity of soil acid-phosphatase and repression of alkaline phosphatase, indicating a stimulatory effect on the microbial population. A soil-free enrichment of microorganisms adapted to heavy-metal and acidic conditions was derived from the TEP-amended soil microcosms using TEP as the sole phosphate source and the selected microbial consortium maintained a high acid-phosphatase activity with repression of alkaline phosphatase. Addition of 5mM zinc to soil-free microcosms had little effect on acid phosphatase but inhibited alkaline phosphatase. One bacterial member from the consortium, identified as Burkholderia cepacia sp., expressed an acid-phosphatase activity uninhibited by high concentrations of zinc and produced a soluble, indigo pigment under phosphate limitation. The pigment was produced in a phosphate-free medium and was not produced in the presence of TEP or phosphate ion, indicative of purple acid-phosphatase types that are pressed by bioavailable phosphate. These results demonstrate that TEP amendment was bioavailable and increased overall phosphatase activity in both soil and soil-free microcosms supporting the possibility of positive outcomes in bioremediation applications. Copyright © 2016. Published by Elsevier Inc.

  18. How steep are the Alps?

    NASA Astrophysics Data System (ADS)

    Robl, Jörg; Prasicek, Günther; Stüwe, Kurt; Hergarten, Stefan

    2014-05-01

    The topography of the European Alps reflects continental collision, crustal thickening and buoyancy driven surface uplift, overprinted by erosional processes. Topographic gradients generally steepen from the valley floors up to about 1500 m - 2000 m followed by an unexpected decrease in slope up to about 2900 m and a further increase to the highest summits of the range. Several studies have interpreted this pattern and the accompanied maximum in the hypsometric curve in terms of either the critical slope stability angle, the prematurity of the Alps caused by recent tectonic uplift, or the effect of the glacial "buzz saw" related to the Pleistocene glaciation cycles. There is consensus that the lithological inventory represents a first order parameter for the steepness of fluvial channels and the angle of hillslopes in steady state and that the response time of a transient landscape is controlled by lithology. In this study we systematically explore the slope-elevation distributions for several hundred continuous domains of the major structural units of the Alps. For this, we apply a novel numerical code to determine the predominant cause for the observed peculiar topography. We compare adjacent alpine domains with contrasting lithology to explore lithological effects on the limiting slope stability angle. We analyze domains with different lithology in the non-glaciated parts of the orogen to highlight the state of maturity related to a recent uplift event. We evaluate the glacial effects on the landscape by the comparison of areas belonging to the same structural units but affected by a variable amount of glacial imprint. The results show that lithology has a major impact on the morphometric characteristics of the European Alps. Adjacent but different structural units show a significant variability in their slope-elevation distributions although they have experienced the same uplift history and the same amount of glacial imprint. This suggests that the response

  19. A beta-N-acetylglucosaminyl phosphate diester residue is attached to the glycosylphosphatidylinositol anchor of human placental alkaline phosphatase: a target of the channel-forming toxin aerolysin.

    PubMed

    Fukushima, Keiko; Ikehara, Yukio; Kanai, Michiko; Kochibe, Naohisa; Kuroki, Masahide; Yamashita, Katsuko

    2003-09-19

    Glycosylphosphatidylinositol (GPI)-anchored proteins are ubiquitous in eukaryotes. The minimum conserved GPI core structure of all GPI-anchored glycans has been determined as EtN-PO4-6Manalpha1-2Manalpha1-6Manalpha1-4GlcN-myo-inositol-PO3H. Human placental alkaline phosphatase (AP) has been reported to be a GPI-anchored membrane protein. AP carries one N-glycan, (NeuAcalpha2-->3)2Gal2GlcNAc2Man3GlcNAc(+/-Fuc)GlcNAc, and a GPI anchor, which contains an ethanolamine phosphate diester group, as a side chain. However, we found that both sialidase-treated soluble AP (sAP) and its GPI-anchored glycan bound to a Psathyrella velutina lectin (PVL)-Sepharose column, which binds beta-GlcNAc residues. PVL binding of asialo-sAP and its GPI-anchored glycan was diminished by digestion with diplococcal beta-N-acetylhexosaminidase or by mild acid treatment. After sequential digestion of asialo-sAP with beta-N-acetylhexosaminidase and acid phosphatase, the elution patterns on chromatofocusing gels were changed in accordance with the negative charges of phosphate residues. Trypsin-digested sAP was analyzed by liquid chromatography/electrospray ionization mass spectrometry, and the structures of two glycopeptides with GPI-anchored glycans were confirmed as peptide-EtN-PO4-6Manalpha1-->2(GlcNAcbeta1-PO4-->6)Manalpha1-6(+/-EtN-PO4-->)Manalpha1-->4GlcN, which may be produced by endo-alpha-glucosaminidase. In addition to AP, GPI-anchored carcinoembryonic antigen, cholinesterase, and Tamm-Horsfall glycoprotein also bound to a PVL-Sepharose column, suggesting that the beta-N-acetylglucosaminyl phosphate diester residue is widely distributed in human GPI-anchored glycans. Furthermore, we found that the beta-N-acetylglucosaminyl phosphate diester residue is important for GPI anchor recognition of aerolysin, a channel-forming toxin derived from Aeromonas hydrophila.

  20. Outer membrane protein e of Escherichia coli K-12 is co-regulated with alkaline phosphatase.

    PubMed

    Tommassen, J; Lugtenberg, B

    1980-07-01

    Outer membrane protein e is induced in wild-type cells, just like alkaline phosphatase and some other periplasmic proteins, by growth under phosphatase limitation. nmpA and nmpB mutants, which synthesize protein e constitutively, are shown also to produce the periplasmic enzyme alkaline phosphatase constitutively. Alternatively, individual phoS, phoT, and phoR mutants as well as pit pst double mutants, all of which are known to produce alkaline phosphatase constitutively, were found to be constitutive for protein e. Also, the periplasmic space of most nmpA mutants and of all nmpB mutants grown in excess phosphate was found to contain, in addition to alkaline phosphatase, at least two new proteins, a phenomenon known for individual phoT and phoR mutants as well as for pit pst double mutants. The other nmpA mutants as well as phoS mutants lacked one of these extra periplasmic proteins, namely the phosphate-binding protein. From these data and from the known positions of the mentioned genes on the chromosomal map, it is concluded that nmpB mutants are identical to phoR mutants. Moreover, some nmpA mutants were shown to be identical to phoS mutants, whereas other nmpA mutants are likely to contain mutations in one of the genes phoS, phoT, or pst.

  1. [Alkaline phosphatase in Amoeba proteus].

    PubMed

    Sopina, V A

    2005-01-01

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

  2. Intracellular accumulation and oligosaccharide processing of alkaline phosphatase under disassembly of the Golgi complex caused by brefeldin A.

    PubMed

    Takami, N; Oda, K; Fujiwara, T; Ikehara, Y

    1990-12-27

    Electron microscopic observations showed that the fungal metabolite brefeldin A caused disassembly of the Golgi complex in human choriocarcinoma cells and accumulation of alkaline phosphatase (ALP) in the endoplasmic reticulum (ER) and nuclear envelope, where ALP was not apparently detectable in control cells. Pulse/chase experiments with [35S]methionine demonstrated that in the control cells, ALP synthesized as a 63-kDa precursor form was rapidly converted to a 66-kDa form, by processing of its N-linked oligosaccharides from the high-mannose type to the complex type, which was expressed on the cell surface after 30 min of chase. In contrast, in the brefeldin-A-treated cells the precursor was gradually converted to a 65-kDa form, slightly smaller than the control mature form, which was not expressed on the cell surface even after a prolonged time of chase. Kinetics of the ALP processing in the brefeldin-A-treated cells demonstrated that the precursor was initially converted to an intermediate form, partially sensitive to endo-beta-N-acetylglucosaminidase H (endo H), then to an endo-H-resistant 65-kDa form. In addition, this form was found to be sensitive to neuraminidase digestion, though its sialylation was not so complete as that of the control mature form. Taken together, these results suggest that under disassembly of the Golgi complex caused by brefeldin A, oligosaccharide-processing enzymes including sialyltransferase, an enzyme in the trans Golgi cisterna(e) and/or the trans Golgi network, might be redistributed into the ER and involved in processing of the oligosaccharides of ALP accumulating there.

  3. In vitro and in vivo evaluation of a new nanocomposite, containing high density polyethylene, tricalcium phosphate, hydroxyapatite, and magnesium oxide nanoparticles.

    PubMed

    Pourdanesh, Fereydoun; Jebali, Ali; Hekmatimoghaddam, Seyedhossein; Allaveisie, Azra

    2014-07-01

    In this study, a new nanocomposite, which contained high density polyethylene (HDPE), tricalcium phosphate (Ca3(PO4)2) nanoparticles (TCP NPs), hydroxyapatite nanoparticles (HA NPs), and magnesium oxide nanoparticles (MgO NPs) was prepared. As in vitro experiment, human osteoblasts (HOB) cells were exposed to pristine HDPE and its nanocomposite for a period of 1, 4, and 7 days at 37 °C, and then different assays were carried out, including osteoblast cell proliferation, Trypan blue staining, cell viability, alkaline phosphatase (ALP), and cell adhesion. Antibacterial property of pristine HDPE and its nanocomposite was evaluated, and also their mechanical properties were measured after 2 and 4 months. As in vivo experiment, pristine HDPE and its nanocomposite were separately implanted on calvarium bone of rabbits, and tissue inflammation and osteogenesis were investigated after 2, 4, and 6 months. In case of HOB cells treated with HDPE or nanocomposite, as incubation time was increased, cell proliferation, live/dead ratio, and cell viability were decreased. But, the ALP activity and cell adhesion of HOB cells which treated with nanocomposite were raised after increase of incubation time. This study demonstrated that although the mechanical properties of nanocomposite were similar to HDPE sheet, but their antibacterial property was not similar. The in vivo experiment showed that both pristine HDPE and its nanocomposite had same inflammation responses. Interestingly, osteogenesis was observed after 2 months at bone/nanocomposite interface, and was highly increased after 4 and 6 months. It must be noted that such pattern was not seen at bone/HDPE interface. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Magnesium substitution in the structure of orthopedic nanoparticles: A comparison between amorphous magnesium phosphates, calcium magnesium phosphates, and hydroxyapatites.

    PubMed

    Nabiyouni, Maryam; Ren, Yufu; Bhaduri, Sarit B

    2015-01-01

    As biocompatible materials, magnesium phosphates have received a lot of attention for orthopedic applications. During the last decade multiple studies have shown advantages for magnesium phosphate such as lack of cytotoxicity, biocompatibility, strong mechanical properties, and high biodegradability. The present study investigates the role of Mg(+2) and Ca(+2) ions in the structure of magnesium phosphate and calcium phosphate nanoparticles. To directly compare the effect of Mg(+2) and Ca(+2) ions on structure of nanoparticles and their biological behavior, three groups of nanoparticles including amorphous magnesium phosphates (AMPs) which release Mg(+2), calcium magnesium phosphates (CMPs) which release Mg(+2) and Ca(+2), and hydroxyapatites (HAs) which release Ca(+2) were studied. SEM, TEM, XRD, and FTIR were used to evaluate the morphology, crystallinity, and chemical properties of the particles. AMP particles were homogeneous nanospheres, whereas CMPs were combinations of heterogeneous nanorods and nanospheres, and HAs which contained heterogeneous nanosphere particles. Cell compatibility was monitored in all groups to determine the cytotoxicity effect of particles on studied MC3T3-E1 preosteoblasts. AMPs showed significantly higher attachment rate than the HAs after 1 day and both AMPs and CMPs showed significantly higher proliferation rate when compared to HAs after 7days. Gene expression level of osteoblastic markers ALP, COL I, OCN, OPN, RUNX2 were monitored and they were normalized to GAPDH housekeeping gene. Beta actin expression level was monitored as the second housekeeping gene to confirm the accuracy of results. In general, AMPs and CMPs showed higher expression level of osteoblastic genes after 7 days which can further confirm the stimulating role of Mg(+2) and Ca(+2) ions in increasing the proliferation rate, differentiation, and mineralization of MC3T3-E1 preosteoblasts. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Phosphate-solubility and phosphatase activity in Gangetic alluvial soil as influenced by organophosphate insecticide residues.

    PubMed

    Majumder, Shyam Prasad; Das, Amal Chandra

    2016-04-01

    An experiment was conducted under laboratory conditions to investigate the effect of four organophosphate insecticides, viz. monocrotophos, profenophos, quinalphos and triazophos at their field application rates (0.75, 1.0, 0.5 and 0.6 kg a.i.ha(-1), respectively), on the growth and activities of phosphate solubilizing microorganisms in relation to availability of insoluble phosphates in the Gangetic alluvial soil of West Bengal, India. The proliferation of phosphate solubilizing microorganisms was highly induced with profenophos (38.3%), while monocrotophos exerted maximum stimulation (20.8%) towards the solubility of insoluble phosphates in soil. The phosphatase activities of the soil (both acid phosphatase and alkaline phosphatase) were significantly increased due to the incorporation of the insecticides in general, and the augmentation was more pronounced with quinalphos (43.1%) followed by profenophos (27.6%) for acid phosphatase, and with monocrotophos (25.2%) followed by profenophos (16.1%) for alkaline phosphatase activity in soil. The total phosphorus was highly retained by triazophos (19.9%) followed by monocrotophos (16.5%), while incorporation of triazophos and quinalphos manifested greater availability of water soluble phosphorus in soil. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. A magnetotelluric feasibility study of the Alps

    NASA Astrophysics Data System (ADS)

    Ritter, O.; Weckmann, U.

    2016-12-01

    The Alps are a famous and extensive mountain range system in central Europe. The mountains were formed as the African and Eurasian tectonic plates collided and they have been a prime target for geological and geophysical investigations since the beginning of modern geosciences. Consequently, the Alps have been investigated with active and passive seismological methods and extensive sets of potential field data exist. Hardly anything is known, however, about the deep electrical conductivity structure, as it has been notoriously difficult to acquire magnetotelluric (MT) data in the Alps. The Alps are densely populated and a lot of infrastructure for tourism has been built over the years. MT measurements, which rely on natural variations of the electromagnetic background fields, are severely hampered by this man-made noise. Here, we report on a feasibility study to acquire MT data in the Alps, where all stations are deployed outside the valleys, on high mountain ranges and alpine pastures. Overall we recorded MT data at 7 stations, along an approximately north-south profile centred on Mayrhofen in the Austrian Alps. The average station spacing was 5 kilometers. The data were processed using robust remote-reference processing and the results clearly show that MT measurements are feasible. We used Mare2DEM for 2D inversion to include a somewhat realistic topography. The 2D section indicates moderate resistivity for the top 2 - 5 km, consistent with the regional geology, which suggests (meta-) sedimentary sequences. From depths of 5 km and below resistivities exceed 5,000 Ohmm. This means we can sense very deep with MT but also, that we should be cautious with an interpretation of this short profile. The data also clearly indicate 3D effects. We therefore propose to deploy an array of stations covering the entire Alps in USArray style, e.g. with a station spacing of approximately 50 km, to derive a 3D model of the deep electrical resistivity structure of the Alps. Such a

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

    PubMed

    Sun, Longhao; He, Xianghui; Liu, Tong

    2014-01-01

    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. 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. 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. 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.

  8. Phosphates behaviours in conversion of FP chlorides

    NASA Astrophysics Data System (ADS)

    Amamoto, I.; Kofuji, H.; Myochin, M.; Takasaki, Y.; Terai, T.

    2009-06-01

    The spent electrolyte of the pyroprocessing by metal electrorefining method should be considered for recycling after removal of fission products (FP) such as, alkali metals (AL), alkaline earth metals (ALE), and/or rare earth elements (REE), to reduce the volume of high-level radioactive waste. Among the various methods suggested for this purpose is precipitation by converting FP from chlorides to phosphates. Authors have been carrying out the theoretical analysis and experiment showing the behaviours of phosphate precipitates so as to estimate the feasibility of this method. From acquired results, it was found that AL except lithium and ALE are unlikely to form phosphate precipitates. However their conversion behaviours including REE were compatible with the theoretical analysis; in the case of LaPO 4 as one of the REE precipitates, submicron-size particles could be observed while that of Li 3PO 4 was larger; the precipitates were apt to grow larger at higher temperature; etc.

  9. Dentition phase and chronological age in relation to gingival crevicular fluid alkaline phosphatase activity in growing subjects.

    PubMed

    Perinetti, Giuseppe; Baccetti, Tiziano; Di Leonardo, Bruno; Di Lenarda, Roberto; Contardo, Luca

    2011-11-01

    Identification of skeletal maturation phases is of primary importance in terms of individual responsiveness to nearly all dentofacial orthopaedic treatments. In this regard, dentition phase and chronological age are still widely used to define the timing of and responsiveness to orthodontic treatments. Recently, gingival crevicular fluid (GCF) alkaline phosphatase (ALP) activity has been shown to be a reliable biomarker of skeletal maturation in growing subjects. Here, for the first time, circumpubertal dentition phases and chronological age were evaluated for correlations with GCF ALP activity, as a biomarker of skeletal maturation. Eighty-five healthy growing subjects (51 females, 34 males; mean age, 11.7±2.3 years) were enrolled into this double-blind, prospective, cross-sectional-design study. Samples of GCF were collected from each subject at the mesial and distal sites of both of the central incisors, at the maxillary and mandibular arches. Their dentition phases were recorded as intermediate mixed, late mixed, or permanent. GCF ALP enzymatic activity was determined spectrophotometrically. The dentition phases showed median GCF ALP activities from 42.0 to 67.5 mU/sample. Although these were slightly greater for the permanent dentition, no significant differences were seen. Also, the chronological age did not correlate significantly with GCF ALP activity, and no significant differences were seen between maxillary and mandibular sites in any of the comparisons. Assessment for treatment timing of dentofacial disharmonies in individual patients that require monitoring of their skeletal maturation phases should not rely on their circumpubertal dentition phase and chronological age. Copyright © 2011 Società Italiana di Ortodonzia SIDO. Published by Elsevier Srl. All rights reserved.

  10. An electrochemical microRNAs biosensor with the signal amplification of alkaline phosphatase and electrochemical-chemical-chemical redox cycling.

    PubMed

    Xia, Ning; Zhang, Youjuan; Wei, Xin; Huang, Yaping; Liu, Lin

    2015-06-09

    MicroRNAs (MiRNAs) have been regarded as clinically important biomarkers and drug discovery targets. In this work, we reported a simple and ultrasensitive electrochemical method for miRNAs detection based on single enzyme amplification and electrochemical-chemical-chemical (ECC) redox cycling. Specifically, upon contact with the target miRNAs, the hairpin structure of biotinylated DNA immobilized on gold electrode was destroyed and the biotin group in DNA was forced away from the electrode surface, allowing for the coupling of streptavidin-conjugated alkaline phosphatase (SA-ALP). Then, ascorbic acid (AA, the enzymatic product of ALP) triggered the ECC redox cycling with ferrocene methanol (FcM) and tris(2-carboxyethyl)phosphine (TCEP) as the redox mediator and the chemical reducing reagent, respectively. The method was more sensitive than that with horseradish peroxidase (HRP) or glucose oxidase (GOx) triggered recycling since one ALP molecule captured by one target miRNA molecule promoted the production of thousands of AA. Analytical merits (e.g., detection limit, dynamic range, specificity, regeneration and reproducibility) were evaluated. The feasibility of the method for analysis of miRNA-21 in human serum has also been demonstrated. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Extragalactic photon-ALP conversion at CTA energies

    DOE PAGES

    Kartavtsev, A.; Raffelt, G.; Vogel, H.

    2017-01-12

    Magnetic fields in extragalactic space between galaxy clusters may induce conversions between photons and axion-like particles (ALPs), thereby shielding the photons from absorption on the extragalactic background light. For TeV gamma rays, the oscillation length (l osc) of the photon-ALP system becomes inevitably of the same order as the coherence length of the magnetic field l and the length over which the field changes significantly (transition length l t) due to refraction on background photons. We derive exact statistical evolution equations for the mean and variance of the photon and ALP transfer functions in the non-adiabatic regime (l osc ~more » l >> l t). We also make analytical predictions for the transfer functions in the quasi-adiabatic regime (l osc ALP masses.« less

  12. Present-day uplift of the western Alps.

    PubMed

    Nocquet, J-M; Sue, C; Walpersdorf, A; Tran, T; Lenôtre, N; Vernant, P; Cushing, M; Jouanne, F; Masson, F; Baize, S; Chéry, J; van der Beek, P A

    2016-06-27

    Collisional mountain belts grow as a consequence of continental plate convergence and eventually disappear under the combined effects of gravitational collapse and erosion. Using a decade of GPS data, we show that the western Alps are currently characterized by zero horizontal velocity boundary conditions, offering the opportunity to investigate orogen evolution at the time of cessation of plate convergence. We find no significant horizontal motion within the belt, but GPS and levelling measurements independently show a regional pattern of uplift reaching ~2.5 mm/yr in the northwestern Alps. Unless a low viscosity crustal root under the northwestern Alps locally enhances the vertical response to surface unloading, the summed effects of isostatic responses to erosion and glaciation explain at most 60% of the observed uplift rates. Rock-uplift rates corrected from transient glacial isostatic adjustment contributions likely exceed erosion rates in the northwestern Alps. In the absence of active convergence, the observed surface uplift must result from deep-seated processes.

  13. Dual spectra band emissive Eu2+/Mn2+ co-activated alkaline earth phosphates for indoor plant growth novel phosphor converted-LEDs.

    PubMed

    Yun, Young Jun; Kim, Jin Kyu; Ju, Ji Young; Choi, Seul Ki; Park, Woon Ik; Suh, Jae Yong; Jung, Ha-Kyun; Kim, Yongseon; Choi, Sungho

    2017-05-10

    This paper reports designing a novel single composition blue/red color illuminating phosphor followed by fabricating "smart" agricultural/horticultural LED lighting. Color-tunable Eu 2+ /Mn 2+ co-activated alkaline earth phosphates, Na(Sr,Ba)PO 4 and Ca 3 Mg 3 (PO 4 ) 4 , are considered, and the stable doping sites for the corresponding activators are identified by using first-principle DFT calculations. We can realize the designated color purity with stable thermal quenching preserved luminescence behavior is induced by the Eu 2+ center positioned at different coordination states with intermixed Sr 2+ /Ba 2+ sites in Na(Sr,Ba)PO 4 hosts. Moreover, we demonstrate that the resultant LED lighting adopting the proposed novel phosphor composition stimulates the enhanced photosynthesis reaction for indoor hydroponics plants, such as oats and onions, which is superior to the narrow line emission band induced by the mixture of conventional red/green/blue LEDs. Thus, using the color-tunable single composition luminescent material may produce an innovative energy-efficient artificial lighting for indoor plant growth.

  14. A sinter-resistant catalyst using an alumina support recycled from AlP fumigation residue: trash to treasure.

    PubMed

    Dong, Jinshi; Wang, Jun; Wang, Jianqiang; Cheng, Guanghao; Huang, Tianming; Shen, Meiqing

    2018-05-07

    Sintering is a long-standing issue especially in high temperature catalytic applications. In this paper, we report an effective method to slow down metal particle migration and coalescence (PMC) by using a thermally stable alumina support. Noteworthily, the alumina sample was developed from AlP fumigation residue, which is a very dangerous substance for living creatures and environment protection. By optimizing the heated hydrolysis and ball-milling conditions, we recycled a phosphate-stabilized alumina material that retained a 117 m 2 g -1 surface area after 1050 °C hydrothermal aging. The catalyst using this newly developed alumina support had Pd dispersion 1.7 times higher than that using a commercial alumina support after aging. The kinetics and XPS experiments showed that phosphate neither participated in the catalytic reaction process nor changed the active sites. This catalyst also exhibited extraordinary water tolerance and durability, making it a promising material in automotive exhaust purification and other catalytic applications.

  15. Alkaline phosphatase as a screening test for osteomalacia.

    PubMed

    Chinoy, Muhammad Amin; Javed, Muhammad Imran; Khan, Alamzeb; Sadruddin, Nooruddin

    2011-01-01

    Vitamin D deficiency remains common in children and adults in Pakistan despite adequate sunlight exposure. Diagnosis in adults is usually delayed and is made following pathological fractures that result in significant morbidity. The objective of this study was to see whether Serum Alkaline Phosphatase levels could be used as a screening test for osteomalacia. The Study was conducted at Fatima Hospital, Baqai Medical University, Gadap, Karachi, between July 2002 and June 2005. Serum calcium levels are commonly used to screen patients suspected of osteomalacia, and raised serum alkaline phosphatase (SALP) is considered a diagnostic finding. We used SALP to screen patients who presented with back or non-specific aches and pain of more than six months duration. Three hundred thirty-four (334) patients were screened of which 116 (35%) had raised SALP. Osteomalacia was diagnosed in 92 (79.3%) of these 116 either by plain radiographs, bone biopsy or isotope bone scan. Fifty-four (53.4%) of the 101 cases had a normal level of serum calcium. Osteomalacia is likely to be missed if only serum calcium is used to screen patients. Serum Alkaline Phosphate should be used as the preferred method for screening these patients.

  16. VSL#3 probiotic upregulates intestinal mucosal alkaline sphingomyelinase and reduces inflammation.

    PubMed

    Soo, I; Madsen, K L; Tejpar, Q; Sydora, B C; Sherbaniuk, R; Cinque, B; Di Marzio, L; Cifone, M Grazia; Desimone, C; Fedorak, R N

    2008-03-01

    Alkaline sphingomyelinase, an enzyme found exclusively in bile and the intestinal brush border, hydrolyzes sphingomyelin into ceramide, sphingosine and sphingosine-1-phosphate, thereby inducing epithelial apoptosis. Reduced levels of alkaline sphingomyelinase have been found in premalignant and malignant intestinal epithelia and in ulcerative colitis tissue. Probiotic bacteria can be a source of sphingomyelinase. To determine the effect of VSL#3 probiotic therapy on mucosal levels of alkaline sphingomyelinase, both in a mouse model of colitis and in patients with ulcerative colitis. Interleukin-10 gene-deficient (IL10KO) and wild type control mice were treated with VSL#3 (10(9) colony-forming units per day) for three weeks, after which alkaline sphingomyelinase activity was measured in ileal and colonic tissue. As well, 15 patients with ulcerative colitis were treated with VSL#3 (900 billion bacteria two times per day for five weeks). Alkaline sphingomyelinase activity was measured through biopsies and comparison of ulcerative colitis disease activity index scores obtained before and after treatment. Lowered alkaline sphingomyelinase levels were seen in the colon (P=0.02) and ileum (P=0.04) of IL10KO mice, as compared with controls. Treatment of these mice with VSL#3 resulted in upregulation of mucosal alkaline sphingomyelinase activity in both the colon (P=0.04) and the ileum (P=0.01). VSL#3 treatment of human patients who had ulcerative colitis decreased mean (+/- SEM) ulcerative colitis disease activity index scores from 5.3+/-1.8946 to 0.70+/-0.34 (P=0.02) and increased mucosal alkaline sphingomyelinase activity. Mucosal alkaline sphingomyelinase activity is reduced in the intestine of IL10KO mice with colitis and in humans with ulcerative colitis. VSL#3 probiotic therapy upregulates mucosal alkaline sphingomyelinase activity.

  17. An Alkaline Phosphatase Paradox in a Shelf Sea

    NASA Astrophysics Data System (ADS)

    Davis, C. E.; Mahaffey, C.

    2016-02-01

    Alkaline phosphatase (AP) is an ubiquitous hydrolytic phosphoenzyme that hydrolyses phosphomonester bonds. In the open ocean, the generally accepted paradigm is that when phosphate concentrations are sufficiently depleted (less than 50 nM), AP is produced by organisms to enable utilisation of dissolved organic phosphorus to meet the phosphorus demands of biological processes such as growth and carbon fixation. At higher phosphate concentrations (greater than 100 nM), AP is repressed implying that the excess product competes for active sites at enzyme surfaces. However, our ongoing work on phosphorus cycling in the Celtic Sea, a temperate shelf sea, has challenged this paradigm. We find elevated rates of AP below the thermocline where phosphate concentrations are greater than 700 nM, and a significant correlation between AP and total dissolved phosphorus. Using enzyme labelled fluorescence (ELF) and particle concentrate bioassays, we show that the AP is associated with large detrital and sinking particulate matter, suggesting that rather than AP being induced by the lack of phosphate, it plays an important role in organic matter cycling in this nitrogen limited environment. At the shelf edge, AP was found to be associated with diatoms, which have been found in culture studies to express AP under silica limitation. Our study highlights the need to consider the environmental conditions under which AP is induced or repressed and presents an opportunity to use AP as an indicator of organic phosphorus recycling in high phosphate environments.

  18. Sphingosine 1-phosphate receptor activation enhances BMP-2-induced osteoblast differentiation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sato, Chieri; Iwasaki, Tsuyoshi, E-mail: tsuyo-i@huhs.ac.jp; Kitano, Sachie

    Highlights: Black-Right-Pointing-Pointer We investigated the role of S1P signaling for osteoblast differentiation. Black-Right-Pointing-Pointer Both S1P and FTY enhanced BMP-2-stimulated osteoblast differentiation by C2C12 cells. Black-Right-Pointing-Pointer S1P signaling enhanced BMP-2-stimulated Smad and ERK phosphorylation by C2C12 cells. Black-Right-Pointing-Pointer MEK/ERK signaling is a pathway underlying S1P signaling for osteoblast differentiation. -- Abstract: We previously demonstrated that sphingosine 1-phosphate (S1P) receptor-mediated signaling induced proliferation and prostaglandin productions by synovial cells from rheumatoid arthritis (RA) patients. In the present study we investigated the role of S1P receptor-mediated signaling for osteoblast differentiation. We investigated osteoblast differentiation using C2C12 myoblasts, a cell line derived from murinemore » satellite cells. Osteoblast differentiation was induced by the treatment of bone morphogenic protein (BMP)-2 in the presence or absence of either S1P or FTY720 (FTY), a high-affinity agonist of S1P receptors. Osteoblast differentiation was determined by osteoblast-specific transcription factor, Runx2 mRNA expression, alkaline phosphatase (ALP) activity and osteocalcin production by the cells. Smad1/5/8 and extracellular signal-regulated kinase (ERK) 1/2 phosphorylation was examined by Western blotting. Osteocalcin production by C2C12 cells were determined by ELISA. Runx2 expression and ALP activity by BMP-2-stimulated C2C12 cells were enhanced by addition of either S1P or FTY. Both S1P and FTY enhanced BMP-2-induced ERK1/2 and Smad1/5/8 phosphorylation. The effect of FTY was stronger than that of S1P. S1P receptor-mediated signaling on osteoblast differentiation was inhibited by addition of mitogen-activated protein kinase/ERK kinase (MEK) 1/2 inhibitor, indicating that the S1P receptor-mediated MEK1/2-ERK1/2 signaling pathway enhanced BMP-2-Smad signaling. These results indicate that S1P

  19. Urbanization and depopulation in the Alps.

    PubMed

    Batzing, W; Perlik, M; Dekleva, M

    1996-11-01

    Demographic developments in the European Alpine region are analyzed over the period 1870-1990. The region is defined as including parts of Germany, France, Italy, Liechtenstein, Austria, Switzerland, and Slovenia. "Studies of growth, stagnation, decline, commune size, and altitude in almost 6,000 communes are presented on three colored maps.... It is apparent that two highly divergent processes are at work and, accordingly, statistical mean values reveal little of importance. Approximately one-half of Alpine Europe is undergoing general economic and demographic growth and has experienced significant increase in population since the end of the agricultural era. This development has taken place primarily in low-lying valleys and basins and in areas bordering the Alps that have good access to transport routes. Tourism is not as widespread as generally assumed and is usually characterized by a punctate pattern. Only in the western part of the Eastern Alps does tourism account for widespread population growth at higher altitudes; elsewhere the Alps have not been affected by modern development and the economy and population are declining, with some areas in danger of becoming completely abandoned. The results challenge the earlier concept of the Alps as a rural region, once populated by peasants, where tourism now plays a major role." (EXCERPT)

  20. Fluorescence turn-on detection of alkaline phosphatase activity based on controlled release of PEI-capped Cu nanoclusters from MnO2 nanosheets.

    PubMed

    Zhang, Yunyi; Li, Yongxin; Zhang, Cuiyun; Zhang, Qingfeng; Huang, Xinan; Yang, Meiding; Shahzad, Sohail Anjum; Lo, Kenneth Kam-Wing; Yu, Cong; Jiang, Shichun

    2017-08-01

    A fluorescence turn-on assay for alkaline phosphatase (ALP) activity is developed through the controlled release of polyethyleneimine-capped copper nanoclusters (PEI-capped CuNCs) from the MnO 2 nanosheets. In an aqueous solution, the positively charged PEI-capped CuNCs could be adsorbed onto the surface of the negatively charged MnO 2 nanosheets. Such adsorption through favorable electrostatic interactions could efficiently quench the nanocluster fluorescence emission via resonance energy transfer from the PEI-capped CuNCs to the MnO 2 nanosheets. 2-Phospho-L-ascorbic acid (AAP) could be hydrolyzed to L-ascorbic acid (AA) in the presence of ALP. AA could reduce MnO 2 into Mn 2+ and trigger the disintegration of the MnO 2 nanosheets. As a result, the CuNCs were released and the quenched fluorescence was recovered efficiently. The detection strategy is simple, inexpensive, sensitive, selective, with low toxicity, and has better biocompatibility. The newly fabricated biosensor for ALP activity will potentially make it a robust candidate for numerous biological and biomedical applications.

  1. Preferable adsorption of phosphate using lanthanum-incorporated porous zeolite: Characteristics and mechanism

    NASA Astrophysics Data System (ADS)

    He, Yinhai; Lin, Hai; Dong, Yingbo; Wang, Liang

    2017-12-01

    The adsorbent, where lanthanum oxide was incorporated onto porous zeolite (La-Z), of preferable adsorption towards phosphate was prepared by hydrothermal synthesis. Based on pH effect results, La-Z would effectively sequestrate phosphate over wider pH range of 3.0-7.0, alkaline conditions were unfavorable for phosphate. The adsorption of phosphate was not significantly influenced by ionic strength and by coexisting anions of chloride, nitrate and sulfate but bicarbonate showed slightly greater negative effects, indicating La-Z possessed highly selectivity to phosphate. Adsorption of phosphate could be well fitted by pseudo-second-order model and the process was mainly controlled by intra-particle diffusion. Equilibrium adsorption demonstrated that Langmuir model was more suitable than Freundlich model for description phosphate adsorption and the adsorption capacity was 17.2 mg P g-1, which exhibited 95% utilization of incorporated La. Over 95% phosphate was eliminated in real effluent treatment when the dose was 2 g L-1. The underlying mechanism for phosphate capture was probed with Zeta potential and X-ray photoelectron spectroscope analysis, and the formation of La-P inner-sphere complexation was testified to be the dominant pathway. All the results suggested that the porous zeolite-supported lanthanum oxide can serve as a promising adsorbent for phosphate removal in realistic application.

  2. Gene Expression of Lytic Endopeptidases AlpA and AlpB from Lysobacter sp. XL1 in Pseudomonads.

    PubMed

    Tsfasman, Irina M; Lapteva, Yulia S; Krasovskaya, Ludmila A; Kudryakova, Irina V; Vasilyeva, Natalia V; Granovsky, Igor E; Stepnaya, Olga A

    2015-01-01

    Development of an efficient expression system for (especially secreted) bacterial lytic enzymes is a complicated task due to the specificity of their action. The substrate for such enzymes is peptidoglycan, the main structural component of bacterial cell walls. For this reason, expression of recombinant lytic proteins is often accompanied with lysis of the producing bacterium. This paper presents data on the construction of an inducible system for expression of the lytic peptidases AlpA and AlpB from Lysobacter sp. XL1 in Pseudomonas fluorescens Q2-87, which provides for the successful secretion of these proteins into the culture liquid. In this system, the endopeptidase gene under control of the T7lac promoter was integrated into the bacterial chromosome, as well as the Escherichia coli lactose operon repressor protein gene. The T7 pol gene under lac promoter control, which encodes the phage T7 RNA polymerase, is maintained in Pseudomonas cells on the plasmids. Media and cultivation conditions for the recombinant strains were selected to enable the production of AlpA and AlpB by a simple purification protocol. Production of recombinant lytic enzymes should contribute to the development of new-generation antimicrobial drugs whose application will not be accompanied by selection of resistant microorganisms. © 2015 S. Karger AG, Basel.

  3. Impact of a Genetically Engineered Bacterium with Enhanced Alkaline Phosphatase Activity on Marine Phytoplankton Communities

    PubMed Central

    Sobecky, P. A.; Schell, M. A.; Moran, M. A.; Hodson, R. E.

    1996-01-01

    An indigenous marine Achromobacter sp. was isolated from coastal Georgia seawater and modified in the laboratory by introduction of a plasmid with a phoA hybrid gene that directed constitutive overproduction of alkaline phosphatase. The effects of this "indigenous" genetically engineered microorganism (GEM) on phosphorus cycling were determined in seawater microcosms following the addition of a model dissolved organic phosphorus compound, glycerol 3-phosphate, at a concentration of 1 or 10 (mu)M. Within 48 h, a 2- to 10-fold increase in the concentration of inorganic phosphate occurred in microcosms containing the GEM (added at an initial density equivalent to 8% of the total bacterial population) relative to controls containing only natural microbial populations, natural populations with the unmodified Achromobacter sp., or natural populations with the Achromobacter sp. containing the plasmid but not the phoA gene. Secondary effects of the GEM on the phytoplankton community were observed after several days, evident as sustained increases in phytoplankton biomass (up to 14-fold) over that in controls. Even in the absence of added glycerol 3-phosphate, a numerically stable GEM population (averaging 3 to 5% of culturable bacteria) was established within 2 to 3 weeks of introduction into seawater. Moreover, alkaline phosphatase activity in microcosms with the GEM was substantially higher than that in controls for up to 25 days, and microcosms containing the GEM maintained the potential for net phosphate accumulation above control levels for longer than 1 month. PMID:16535222

  4. Age and Prematurity of the Alps Derived from Topography

    NASA Astrophysics Data System (ADS)

    Hergarten, S.; Wagner, T.; Stüwe, K.

    2010-09-01

    The European Alps are one of the best studied mountain ranges on Earth, but yet the age of their topography is almost unknown. Even their relative stage of evolution is unclear: Are the Alps still growing, in a steady state or already decaying, and is there a significant difference between Western and Eastern Alps? Using a new geomorphic parameter we analyze the topography of the Alps and provide one of the first quantitative constraints demonstrating that the range is still in its infancy: In contrast to several other mountain ranges, the Alps have still more than half of their evolution to a geomorphic steady state to go. Combining our results with sediment budget data from the surrounding basins we infer that the formation of the present topography began only 5-6 million years ago. Our results question the apparent consensus that the topographic evolution is distributed over much of the Miocene and might give new impulses to the reconstruction of paleoclimate in Central Europe.

  5. Characterization of iron-phosphate-silicate chemical garden structures.

    PubMed

    Barge, Laura M; Doloboff, Ivria J; White, Lauren M; Stucky, Galen D; Russell, Michael J; Kanik, Isik

    2012-02-28

    Chemical gardens form when ferrous chloride hydrate seed crystals are added or concentrated solutions are injected into solutions of sodium silicate and potassium phosphate. Various precipitation morphologies are observed depending on silicate and phosphate concentrations, including hollow plumes, bulbs, and tubes. The growth of precipitates is controlled by the internal osmotic pressure, fluid buoyancy, and membrane strength. Additionally, rapid bubble-led growth is observed when silicate concentrations are high. ESEM/EDX analysis confirms compositional gradients within the membranes, and voltage measurements across the membranes during growth show a final potential of around 150-200 mV, indicating that electrochemical gradients are maintained across the membranes as growth proceeds. The characterization of chemical gardens formed with iron, silicate, and phosphate, three important components of an early earth prebiotic hydrothermal system, can help us understand the properties of analogous structures that likely formed at submarine alkaline hydrothermal vents in the Hadean-structures offering themselves as the hatchery of life. © 2011 American Chemical Society

  6. Selective flotation of phosphate minerals with hydroxamate collectors

    DOEpatents

    Miller, Jan D.; Wang, Xuming; Li, Minhua

    2002-01-01

    A method is disclosed for separating phosphate minerals from a mineral mixture, particularly from high-dolomite containing phosphate ores. The method involves conditioning the mineral mixture by contacting in an aqueous in environment with a collector in an amount sufficient for promoting flotation of phosphate minerals. The collector is a hydroxamate compound of the formula; ##STR1## wherein R is generally hydrophobic and chosen such that the collector has solubility or dispersion properties it can be distributed in the mineral mixture, typically an alkyl, aryl, or alkylaryl group having 6 to 18 carbon atoms. M is a cation, typically hydrogen, an alkali metal or an alkaline earth metal. Preferably, the collector also comprises an alcohol of the formula, R'--OH wherein R' is generally hydrophobic and chosen such that the collector has solubility or dispersion properties so that it can be distributed in the mineral mixture, typically an alkyl, aryl, or alkylaryl group having 6 to 18 carbon atoms.

  7. Coordinate responses to alkaline pH stress in budding yeast

    PubMed Central

    Serra-Cardona, Albert; Canadell, David; Ariño, Joaquín

    2015-01-01

    Alkalinization of the medium represents a stress condition for the budding yeast Saccharomyces cerevisiae to which this organism responds with profound remodeling of gene expression. This is the result of the modulation of a substantial number of signaling pathways whose participation in the alkaline response has been elucidated within the last ten years. These regulatory inputs involve not only the conserved Rim101/PacC pathway, but also the calcium-activated phosphatase calcineurin, the Wsc1-Pkc1-Slt2 MAP kinase, the Snf1 and PKA kinases and oxidative stress-response pathways. The uptake of many nutrients is perturbed by alkalinization of the environment and, consequently, an impact on phosphate, iron/copper and glucose homeostatic mechanisms can also be observed. The analysis of available data highlights cases in which diverse signaling pathways are integrated in the gene promoter to shape the appropriate response pattern. Thus, the expression of different genes sharing the same signaling network can be coordinated, allowing functional coupling of their gene products. PMID:28357292

  8. Medium pH, carbon and nitrogen concentrations modulate the phosphate solubilization efficiency of Penicillium purpurogenum through organic acid production.

    PubMed

    Scervino, J M; Papinutti, V L; Godoy, M S; Rodriguez, M A; Della Monica, I; Recchi, M; Pettinari, M J; Godeas, A M

    2011-05-01

    To study phosphate solubilization in Penicillium purpurogenum as function of medium pH, and carbon and nitrogen concentrations. Tricalcium phosphate (CP) solubilization efficiency of P. purpurogenum was evaluated at acid or alkaline pH using different C and N sources. Glucose- and (NH(4) )(2) SO(4) -based media showed the highest P solubilization values followed by fructose. P. purpurogenum solubilizing ability was higher in cultures grown at pH 6·5 than cultures at pH 8·5. Organic acids were detected in both alkaline and neutral media, but the relative percentages of each organic acid differed. Highest P release coincided with the highest organic acids production peak, especially gluconic acid. When P. purpurogenum grew in alkaline media, the nature and concentration of organic acids changed at different N and C concentrations. A factorial categorical experimental design showed that the highest P-solubilizing activity, coinciding with the highest organic acid production, corresponded to the highest C concentration and lowest N concentration. The results described in the present study show that medium pH and carbon and nitrogen concentrations modulate the P solubilization efficiency of P. purpurogenum through the production of organic acids and particularly that of gluconic acid. In the P solubilization optimization studies, glucose and (NH(4) )(2) SO(4) as C and N sources allowed a higher solubilization efficiency at high pH. This organism is a potentially proficient soil inoculant, especially in P-poor alkaline soils where other P solubilizers fail to release soluble P. Further work is necessary to elucidate whether these results can be extrapolated to natural soil ecosystems, where different pH values are present. Penicillium purpurogenum could be used to develop a bioprocess for the manufacture of phosphatic fertilizer with phosphate calcium minerals. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  9. MAPK Signaling Pathway Alters Expression of Midgut ALP and ABCC Genes and Causes Resistance to Bacillus thuringiensis Cry1Ac Toxin in Diamondback Moth

    PubMed Central

    Wu, Qingjun; Wang, Shaoli; Xie, Wen; Zhu, Xun; Baxter, Simon W.; Zhou, Xuguo; Jurat-Fuentes, Juan Luis; Zhang, Youjun

    2015-01-01

    Insecticidal crystal toxins derived from the soil bacterium Bacillus thuringiensis (Bt) are widely used as biopesticide sprays or expressed in transgenic crops to control insect pests. However, large-scale use of Bt has led to field-evolved resistance in several lepidopteran pests. Resistance to Bt Cry1Ac toxin in the diamondback moth, Plutella xylostella (L.), was previously mapped to a multigenic resistance locus (BtR-1). Here, we assembled the 3.15 Mb BtR-1 locus and found high-level resistance to Cry1Ac and Bt biopesticide in four independent P. xylostella strains were all associated with differential expression of a midgut membrane-bound alkaline phosphatase (ALP) outside this locus and a suite of ATP-binding cassette transporter subfamily C (ABCC) genes inside this locus. The interplay between these resistance genes is controlled by a previously uncharacterized trans-regulatory mechanism via the mitogen-activated protein kinase (MAPK) signaling pathway. Molecular, biochemical, and functional analyses have established ALP as a functional Cry1Ac receptor. Phenotypic association experiments revealed that the recessive Cry1Ac resistance was tightly linked to down-regulation of ALP, ABCC2 and ABCC3, whereas it was not linked to up-regulation of ABCC1. Silencing of ABCC2 and ABCC3 in susceptible larvae reduced their susceptibility to Cry1Ac but did not affect the expression of ALP, whereas suppression of MAP4K4, a constitutively transcriptionally-activated MAPK upstream gene within the BtR-1 locus, led to a transient recovery of gene expression thereby restoring the susceptibility in resistant larvae. These results highlight a crucial role for ALP and ABCC genes in field-evolved resistance to Cry1Ac and reveal a novel trans-regulatory signaling mechanism responsible for modulating the expression of these pivotal genes in P. xylostella. PMID:25875245

  10. Orbital fluid shear stress promotes osteoblast metabolism, proliferation and alkaline phosphates activity in vitro

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Aisha, M.D.; Nor-Ashikin, M.N.K.; DDH, Universiti Teknologi MARA, ShahAlam 40450, Selangor

    Prolonged disuse of the musculoskeletal system is associated with reduced mechanical loading and lack of anabolic stimulus. As a form of mechanical signal, the multidirectional orbital fluid shear stress transmits anabolic signal to bone forming cells in promoting cell differentiation, metabolism and proliferation. Signals are channeled through the cytoskeleton framework, directly modifying gene and protein expression. For that reason, we aimed to study the organization of Normal Human Osteoblast (NHOst) cytoskeleton with regards to orbital fluid shear (OFS) stress. Of special interest were the consequences of cytoskeletal reorganization on NHOst metabolism, proliferation, and osteogenic functional markers. Cells stimulated at 250more » RPM in a shaking incubator resulted in the rearrangement of actin and tubulin fibers after 72 h. Orbital shear stress increased NHOst mitochondrial metabolism and proliferation, simultaneously preventing apoptosis. The ratio of RANKL/OPG was reduced, suggesting that orbital shear stress has the potential to inhibit osteoclastogenesis and osteoclast activity. Increase in ALP activity and OCN protein production suggests that stimulation retained osteoblast function. Shear stress possibly generated through actin seemed to hold an anabolic response as osteoblast metabolism and functional markers were enhanced. We hypothesize that by applying orbital shear stress with suitable magnitude and duration as a non-drug anabolic treatment can help improve bone regeneration in prolonged disuse cases. - Highlights: • OFS stress transmits anabolic signals to osteoblasts. • Actin and tubulin fibers are rearranged under OFS stress. • OFS stress increases mitochondrial metabolism and proliferation. • Reduced RANKL/OPG ratio in response to OFS inhibits osteoclastogenesis. • OFS stress prevents apoptosis and stimulates ALP and OCN.« less

  11. The XMAP215 Ortholog Alp14 Promotes Microtubule Nucleation in Fission Yeast.

    PubMed

    Flor-Parra, Ignacio; Iglesias-Romero, Ana Belén; Chang, Fred

    2018-06-04

    The organization and number of microtubules (MTs) in a cell depend on the proper regulation of MT nucleation. Currently, the mechanism of nucleation is the most poorly understood aspect of MT dynamics. XMAP215/chTOG/Alp14/Stu2 proteins are MT polymerases that stimulate MT polymerization at MT plus ends by binding and releasing tubulin dimers. Although these proteins also localize to MT organizing centers and have nucleating activity in vitro, it is not yet clear whether these proteins participate in MT nucleation in vivo. Here, we demonstrate that in the fission yeast Schizosaccharomyces pombe, the XMAP215 ortholog Alp14 is critical for efficient MT nucleation in vivo. In multiple assays, loss of Alp14 function led to reduced nucleation rate and numbers of interphase MT bundles. Conversely, activation of Alp14 led to increased nucleation frequency. Alp14 associated with Mto1 and γ-tubulin complex components, and artificially targeting Alp14 to the γ-tubulin ring complexes (γ-TuRCs) stimulated nucleation. In imaging individual nucleation events, we found that Alp14 transiently associated with a γ-tubulin particle shortly before the appearance of a new MT. The transforming acidic coiled-coil (TACC) ortholog Alp7 mediated the localization of Alp14 at nucleation sites but not plus ends, and was required for efficient nucleation but not for MT polymerization. Our findings provide the strongest evidence to date that Alp14 serves as a critical MT nucleation factor in vivo. We suggest a model in which Alp14 associates with the γ-tubulin complex in an Alp7-dependent manner to facilitate the assembly or stabilization of the nascent MT. Copyright © 2018 Elsevier Ltd. All rights reserved.

  12. Enhanced attenuation of septic system phosphate in noncalcareous sediments.

    PubMed

    Robertson, W D

    2003-01-01

    Review of phosphate behavior in four mature septic system plumes on similar textured sand has revealed a strong correlation between carbonate mineral content and phosphate concentrations. A plume on calcareous sand (Cambridge site, 27 wt % CaCO3 equiv.) has proximal zone PO4 concentrations (4.8 mg/L P average) that are about 75% of the septic tank effluent value, whereas three plumes on noncalcareous sand (Muskoka, L. Joseph, and Nobel sites, <1 wt % CaCO3 equiv.) have proximal zone phosphate concentrations (<0.1 mg/L P) that are consistently less than 2% of the effluent values. Phosphate attenuation at the noncalcareous sites appears to be an indirect result of the development of acidic conditions (site average pH 3.5 to 5.9) and elevated Al concentrations (up to 24 mg/L), which subsequently causes the precipitation of Al-P minerals such as variscite (AlPO4 x 2H2O). This is supported by scanning electron microscope analyses, which show the widespread occurrence of (Al+P)--rich secondary mineral coatings on sand grains below the infiltration beds. All of these septic systems are more than 10 years old, indicating that these attenuation reactions have substantial longevity. A field lysimeter experiment demonstrated that this reaction sequence can be readily incorporated into engineered waste water treatment systems. We feel this important P removal mechanism has not been adequately recognized, particularly for its potential significance in reducing P loading from septic systems in lakeshore environments.

  13. An exploratory analysis of alkaline phosphatase, lactate dehydrogenase, and prostate-specific antigen dynamics in the phase 3 ALSYMPCA trial with radium-223

    PubMed Central

    Sartor, O.; Coleman, R. E.; Nilsson, S.; Heinrich, D.; Helle, S. I.; O’Sullivan, J. M.; Vogelzang, N. J.; Bruland, Ø.; Kobina, S.; Wilhelm, S.; Xu, L.; Shan, M.; Kattan, M. W.; Parker, C.

    2017-01-01

    Background Baseline clinical variables are prognostic for overall survival (OS) in patients with castration-resistant prostate cancer (CRPC). Their prognostic and predictive value with agents targeting bone metastases, such as radium-223, is not established. Patients and methods The radium-223 ALSYMPCA trial enrolled patients with CRPC and symptomatic bone metastases. Prognostic potential of baseline variables was assessed using Cox models. Percentage changes in biomarker levels from baseline were evaluated during the trial period; changes from baseline to week 12 were evaluated for association with OS and surrogacy. Results Eastern Cooperative Oncology Group performance status, total alkaline phosphatase (tALP), lactate dehydrogenase (LDH), and prostate-specific antigen (PSA) at baseline were associated with OS (P ≤ 0.0003) in the intent-to-treat population (radium-223, N = 614; placebo, N = 307). tALP declined from baseline within 4 weeks after beginning radium-223, by week 12 declining in 87% of radium-223 and 23% of placebo patients (P < 0.001). LDH declined in 51% and 34% (P = 0.003), whereas PSA declined in 27% and 14% (P = 0.160). Mean tALP change from baseline was 32.2% decrease with radium-223 and 37.2% increase with placebo. Radium-223 patients with tALP decline from baseline to week 12 (confirmed ≥3 weeks from week 12) had 55% lower risk of death (hazard ratio = 0.45; 95% CI 0.34–0.61) versus those with no confirmed tALP decline. Proportional treatment effect (PTE) values for tALP, LDH, and PSA changes from baseline at week 12 as OS surrogate markers were 0.34 (95% CI: 0–0.746), 0.07 (95% CI: 0–0.211), and 0 (95% CI: 0–0.082), respectively. Conclusions Significant tALP declines (versus placebo) occurred as early as 4 weeks after beginning radium-223 therapy. tALP or LDH declines at 12 weeks correlated with longer OS, but did not meet statistical surrogacy requirements. Dynamic changes in tALP and LDH during

  14. New Zealand Southern Alps

    NASA Image and Video Library

    2001-06-20

    This anaglyph from the MISR instrument aboard NASA Terra spacecraft shows the rugged Southern Alps extending some 650 kilometers along the western side of New Zealand South Island. 3D glasses are necessary to view this image.

  15. Phosphate homeostasis in Bartter syndrome: a case-control study.

    PubMed

    Bettinelli, Alberto; Viganò, Cristina; Provero, Maria Cristina; Barretta, Francesco; Albisetti, Alessandra; Tedeschi, Silvana; Scicchitano, Barbara; Bianchetti, Mario G

    2014-11-01

    Bartter patients may be hypercalciuric. Additional abnormalities in the metabolism of calcium, phosphate, and calciotropic hormones have occasionally been reported. The metabolism of calcium, phosphate, and calciotropic hormones was investigated in 15 patients with Bartter syndrome and 15 healthy subjects. Compared to the controls, Bartter patients had significantly reduced plasma phosphate {mean [interquartile range]:1.29 [1.16-1.46] vs. 1.61 [1.54-1.67] mmol/L} and maximal tubular phosphate reabsorption (1.16 [1.00-1.35] vs. 1.41 [1.37-1.47] mmol/L) and significantly increased parathyroid hormone (PTH) level (6.1 [4.5-7.7] vs. 2.8 [2.2-4.4] pmol/L). However, patients and controls did not differ in blood calcium, 25-hydroxyvitamin D, alkaline phosphatase, and osteocalcin levels. In patients, an inverse correlation (P < 0.05) was noted between total plasma calcium or glomerular filtration rate and PTH concentration. A positive correlation was also noted between PTH and osteocalcin concentrations (P < 0.005), as well as between chloriduria or natriuria and phosphaturia (P < 0.001). No correlation was noted between calciuria and PTH concentration or between urinary or circulating phosphate and PTH. The results of this study demonstrate a tendency towards renal phosphate wasting and elevated circulating PTH levels in Bartter patients.

  16. Two-Dimensional Magnesium Phosphate Nanosheets Form Highly Thixotropic Gels That Up-Regulate Bone Formation.

    PubMed

    Laurenti, Marco; Al Subaie, Ahmed; Abdallah, Mohamed-Nur; Cortes, Arthur R G; Ackerman, Jerome L; Vali, Hojatollah; Basu, Kaustuv; Zhang, Yu Ling; Murshed, Monzur; Strandman, Satu; Zhu, Julian; Makhoul, Nicholas; Barralet, Jake E; Tamimi, Faleh

    2016-08-10

    Hydrogels composed of two-dimensional (2D) nanomaterials have become an important alternative to replace traditional inorganic scaffolds for tissue engineering. Here, we describe a novel nanocrystalline material with 2D morphology that was synthesized by tuning the crystallization of the sodium-magnesium-phosphate system. We discovered that the sodium ion can regulate the precipitation of magnesium phosphate by interacting with the crystal's surface causing a preferential crystal growth that results in 2D morphology. The 2D nanomaterial gave rise to a physical hydrogel that presented extreme thixotropy, injectability, biocompatibility, bioresorption, and long-term stability. The nanocrystalline material was characterized in vitro and in vivo and we discovered that it presented unique biological properties. Magnesium phosphate nanosheets accelerated bone healing and osseointegration by enhancing collagen formation, osteoblasts differentiation, and osteoclasts proliferation through up-regulation of COL1A1, RunX2, ALP, OCN, and OPN. In summary, the 2D magnesium phosphate nanosheets could bring a paradigm shift in the field of minimally invasive orthopedic and craniofacial interventions because it is the only material available that can be injected through high gauge needles into bone defects in order to accelerate bone healing and osseointegration.

  17. Analysis of human bone alkaline phosphatase isoforms: comparison of isoelectric focusing and ion-exchange high-performance liquid chromatography.

    PubMed

    Sharp, Christopher A; Linder, Cecilia; Magnusson, Per

    2007-04-01

    Several isoforms of alkaline phosphatase (ALP) can be identified in human tissues and serum after separation by anion-exchange HPLC and isoelectric focusing (IEF). We purified four soluble bone ALP (BALP) isoforms (B/I, B1x, B1 and B2) from human SaOS-2 cells, determined their specific pI values by broad range IEF (pH 3.5-9.5), compared these with commercial preparations of bone, intestinal and liver ALPs and established the effects of neuraminidase and wheat germ lectin (WGA) on enzyme activity. Whilst the isoforms B1x (pI=4.48), B1 (pI=4.32) and B2 (pI=4.12) resolved as well-defined bands, B/I resolved as a complex (pI=4.85-6.84). Neuraminidase altered the migration of all BALP isoforms to pI=6.84 and abolished their binding to the anion-exchange matrix, but increased their enzymatic activities by 11-20%. WGA precipitated the BALP isoforms in IEF gels and the HPLC column and attenuated their enzymatic activities by 54-73%. IEF resolved the commercial BALP into 2 major bands (pI=4.41 and 4.55). Migration of BALP isoforms is similar in IEF and anion-exchange HPLC and dependent on sialic acid content. HPLC is preferable in smaller scale research applications where samples containing mixtures of BALP isoforms are analysed. Circulating liver ALP (pI=3.85) can be resolved from BALP by either method. IEF represents a simpler approach for routine purposes even though some overlapping of the isoforms may occur.

  18. Theoretical Studies of the Spin Hamiltonian Parameters and Local Distortions for Cu2+ in Alkaline Earth Lead Zinc Phosphate Glasses

    NASA Astrophysics Data System (ADS)

    Wang, Bo-Kun; Wu, Shao-Yi; Yuan, Zi-Yi; Liu, Zi-Xuan; Jiang, Shi-Xin; Liu, Zheng; Yao, Zi-Jian; Teng, Bao-Hua; Wu, Ming-He

    2016-08-01

    The spin Hamiltonian parameters and local structures are theoretically studied for Cu2+-doped alkaline earth lead zinc phosphate (RPPZ, R=Mg, Ca, Sr, and Ba) glasses based on the high-order perturbation calculations for a tetragonally elongated octahedral 3d9 cluster. The relative elongation ratios are found to be ρ≈3.2%, 4.4%, 4.6%, and 3.3% for R=Mg, Ca, Sr, and Ba, respectively, because of the Jahn-Teller effect. The whole decreasing crystal-field strength Dq and orbital reduction factor k from Mg to Sr are ascribed to the weakening electrostatic coulombic interactions and the increasing probability of productivity of nonbridge oxygen (and hence increasing Cu2+-O2- electron cloud admixtures) under PbO addition, respectively, with increasing alkali earth ionic radius. The anomalies (the largest Dq and the next highest k among the systems) for R=Ba are attributed to the cross linkage of this large cation in the network. The overall increasing order (Mg≤Ba

  19. Bismuth citrate in the quantification of inorganic phosphate and its utility in the determination of membrane-bound phosphatases.

    PubMed

    Cariani, L; Thomas, L; Brito, J; del Castillo, J R

    2004-01-01

    This paper describes a rapid and sensitive method to determine inorganic phosphate, even in the presence of labile organic phosphate compounds and large quantities of proteins. The method eliminates the use of sodium arsenite, a highly toxic compound, substituting bismuth citrate for it to stabilize the phosphomolybdic acid complex formed during the interaction of inorganic phosphate and molybdate reduced by ascorbic acid. This method has also been adapted to microplates and has been used to determine the activities of Na/K ATPase and alkaline phosphatase of intestinal basolateral and luminal plasma membranes.

  20. SALMON-TRINITY ALPS WILDERNESS, CALIFORNIA.

    USGS Publications Warehouse

    Hotz, Preston E.; Thurber, Horace K.

    1984-01-01

    The Salmon-Trinity Alps Wilderness in the Klamath Mountains province occupies an area of about 648 sq mi in parts of Trinity, Siskiyou, and Humboldt Counties, northwestern California. As a result of field studies it was determined that the Salmon-Trinity Alps Wilderness has an area with substantiated potential for gold resources in known lode deposits. Small amounts of quicksilver have been produced from one mine but there is little promise for the discovery of additional mercury resources. Geochemical sampling showed that anomalously high amounts of several other metals occur in a few places, but there is little promise for the discovery of energy or mineral resources other than mercury and gold.

  1. Fission yeast Alp14 is a dose-dependent plus end–tracking microtubule polymerase

    PubMed Central

    Al-Bassam, Jawdat; Kim, Hwajin; Flor-Parra, Ignacio; Lal, Neeraj; Velji, Hamida; Chang, Fred

    2012-01-01

    XMAP215/Dis1 proteins are conserved tubulin-binding TOG-domain proteins that regulate microtubule (MT) plus-end dynamics. Here we show that Alp14, a XMAP215 orthologue in fission yeast, Schizosaccharomyces pombe, has properties of a MT polymerase. In vivo, Alp14 localizes to growing MT plus ends in a manner independent of Mal3 (EB1). alp14-null mutants display short interphase MTs with twofold slower assembly rate and frequent pauses. Alp14 is a homodimer that binds a single tubulin dimer. In vitro, purified Alp14 molecules track growing MT plus ends and accelerate MT assembly threefold. TOG-domain mutants demonstrate that tubulin binding is critical for function and plus end localization. Overexpression of Alp14 or only its TOG domains causes complete MT loss in vivo, and high Alp14 concentration inhibits MT assembly in vitro. These inhibitory effects may arise from Alp14 sequestration of tubulin and effects on the MT. Our studies suggest that Alp14 regulates the polymerization state of tubulin by cycling between a tubulin dimer–bound cytoplasmic state and a MT polymerase state that promotes rapid MT assembly. PMID:22696680

  2. Any Light Particle Search (ALPS)

    NASA Astrophysics Data System (ADS)

    Spector, Aaron; Any Light Particle Search (ALPS) Collaboration

    2016-03-01

    High power laser fields enabled by technologies developed for ground-based gravitational-wave observatories open up new opportunities for fundamental physics studies. One of these options is the search for axions and axion-like particles in a pure laboratory experiment. The axion is a solution to the strong CP-problem and a potential dark matter candidate. The axion has also been proposed as an additional channel to cool stars as well as a potential explanation for the TeV transparency problem. The German-US ALPS collaboration is setting up a light-shining-through-walls (LSW) experiment at DESY. LSW experiments are based on the simple idea that a high power laser field traversing a static magnetic field will transform partly into a relativistic axion field. This axion field will travel through an opaque wall into a second static magnetic field region where it turns partly back into an electromagnetic wave field with the same frequency as the laser. The ALPS collaboration is working towards a large scale LSW experiment at DESY in Hamburg, Germany. I will report on the status of the ALPS experiment. This work is supported by the Deutsche Forschungsgemeinschaft, PRISMA, the Helmholtz Association, the National Science Foundation and the Heising-Simons Foundation.

  3. Scanning electron microscopic and X-ray micro analysis on tooth enamel exposed to alkaline agents.

    PubMed

    Taubee, Fabian; Steiniger, Frank; Nietzsche, Sandor; Norén, Jörgen G

    2010-01-01

    The background of this study comprises two clinical cases, where patients exposed to aerosols of an alkaline and surface active cleaning agent developed loss of enamel substance on their teeth, further resulting in loss of teeth and partially destroyed soft tissues. The alkaline cleaning agent consisted of potassium hydroxide and various surfactants. The purpose of this study was to investigate possible changes in morphology and composition in human teeth enamel exposed to alkaline solutions, by means of X-ray micro analysis (XRMA), FTIR-spectroscopic analyses and scanning electron microscopy (SEM). Extracted premolars, exposed to potassium hydroxide solutions and alkaline cleaning solution,were analyzed by means of XRMA and SEM. Enamel powder, exposed to cleaning solution, was analyzed by means of FTIR. The SEM analysis revealed an increased porosity of the enamel surface and partially loss of enamel substance after exposure to alkaline solutions. The XRMA analyses revealed a decrease in carbon concentration while phosphorous and calcium showed no marked changes. The FTIR analyses showed no significant changes in peak heights or peak positions for phosphate, carbonate or hydroxide. It was concluded that human teeth enamel exposed to alkaline solutions showed loss of organic substance, marked pores in enamel surface and loss of substance in the enamel surface.

  4. Effects of lanthanum carbonate and calcium carbonate on fibroblast growth factor 23 and hepcidin levels in chronic hemodialysis patients.

    PubMed

    Chang, Yu-Ming; Tsai, Shih-Ching; Shiao, Chih-Chung; Liou, Hung-Hsiang; Yang, Chuan-Lan; Tung, Nai-Yu; Hsu, Kua-Sui; Chen, I-Ling; Liu, Mei-Chyn; Kao, Jsun-Liang; Jhen, Rong-Na; Huang, Ya-Ting

    2017-10-01

    Phosphate binders have an impact on fibroblast growth factor 23 (FGF23); however, the effect of phosphate binders on serum hepcidin has not been explored. We conducted a 24-week multicenter randomized controlled trial to investigate the effects of lanthanum carbonate or calcium carbonate monotherapy on serum phosphate, FGF23, and hepcidin levels in chronic hemodialysis patients. Forty-six patients were recruited, and daily dietary phosphorus was controlled between 600-800 mg. Serum calcium, phosphate, albumin, alkaline phosphatase (ALP), FGF23, intact parathyroid hormone (iPTH), hepcidin, high-sensitivity CRP (hsCRP), 25(OH)D, 1,25(OH) 2 D, fetuin-A, and osteopontin were checked as scheduled. Twenty-five patients completed the study. Mean serum FGF23 level was significantly decreased after a 24-week treatment with lanthanum (8677.5 ± 7490.0 vs. 4692.8 ± 5348.3 pg/mL, p = 0.013, n = 13), but not with calcium (n = 12). The reduction of serum hepcidin in lanthanum group was positively correlated with the decrement of serum phosphate (r = 0.631, p = 0.021) and serum hsCRP (r = 0.670, p = 0.012) levels, respectively. Serum ALP, iPTH, vitamin D, fetuin-A, and osteopontin revealed no significant inter- or intragroup differences. In summary, a decrease in serum FGF23 levels and a trend of decline in hepcidin levels were observed only in lanthanum group.

  5. [Influence of cobalt-chromium alloy ceramics crown on aspartate transaminase and alkaline phosphatase of gingival crevicular fluid].

    PubMed

    Miao, Yu; Liu, Ling-Jun; Zhang, Xiao-Min; Li, Li

    2010-12-01

    The purpose of this article was to evaluate the influence of cobalt-chromium (Co-Cr) alloy as the material of inner crown on periodontal tissue through detecting the weight of diseased teeth and the concentration of aspartate transaminase (AST) and alkaline phosphatase (ALP) of the gingival crevicular fluid (GCF) after Co-Cr alloy ceramics repairing. In this study, thirty cases of clinical diseased teeth were chosen from thirty patients based on patients' consent. Each tooth conformed to the inclusion criteria. All of the thirty teeth were repaired with Co-Cr alloy ceramics according to the indications. Then GCF of each diseased tooth was collected and weighed at the time of the day before prosthesis, the first and third month after repairing respectively in order to detect the concentration of AST and ALP. Furthermore, comparative analysis for different periods was performed after the data statistics. To the weight of GCF and the concentration of AST after the respective comparison of three indexes which was of different periods, numerous of each index increased in accordance with the rule of preoperation, postoperative first month, and postoperative third month (P < 0.05). To the concentration of ALP at three time points, the compared results showed that the preoperative numerous was less than that of third month after operation and there was significant difference between them (P < 0.05). To the weight of GCF and the concentration of AST and ALP, after the respective comparison of three indexes which was the diseased teeth and the contralateral teeth with the same name in the periods of pre-operation, all the differences had no statistical significance (P > 0.05), but all the differences had statistical significance in the periods of postoperative third month (P < 0.05). During the next three months after operation, GCF weight, concentration of AST and ALP of diseased teeth was gradually increased after their Co-Cr alloy ceramics crown repairing. This increase

  6. Gadolinium and didymium (praseodymium/neodymium) cations as capture agents in lightmicroscopical histochemistry of acid and alkaline phosphatase.

    PubMed

    Halbhuber, K J; Zimmermann, N

    1987-01-01

    In previous papers, cerium and lanthanum based methods for light-microscopical detection of acid and alkaline phosphatase activity were proposed. In this paper, the usefulness of other lanthanide cations such as gadolinium and praseodymium/neodymium cations as capture agents in phosphatase histochemistry is tested. It is evident that phosphate ions were sufficiently trapped by these cations. According to the lead and silver multistep procedures earlier described it is possible to visualize alkaline phosphatase activity in the brush borders of the intestine or kidney as well as acid phosphatase activity in the lysosomes. These methods can be recommended.

  7. Evaluation of alkaline electrolyzed water to replace traditional phosphate enhancement solutions: Effects on water holding capacity, tenderness, and sensory characteristics.

    PubMed

    Rigdon, Macc; Hung, Yen-Con; Stelzleni, Alexander M

    2017-01-01

    Sixty-four pork loins were randomly assigned to one of four treatments to evaluate the use of alkaline electrolyzed reduced water as a replacement for traditional enhancement solutions. Treatments included: alkaline electrolyzed reduced water (EOH; pH≈11.5), EOH plus 2.5% potassium-lactate (EOK), industry standard (IS; 0.35% sodium tri-polyphosphate, 0.14% sodium chloride, 2.5% potassium-lactate), and no enhancement (CON). After enhancement (targeting 110%) and rest period, chops were cut (2.54-cm) to test treatment effects on water holding capacity, Warner-Bratzler shear force (WBSF), and sensory attributes. Despite its alkaline nature EOH chops exuded more water (P<0.05) than EOK, IS, or CON chops. Control chops were similar (P>0.05) to EOK, however CON and EOK both lost more moisture (P<0.05) than IS. The use of alkaline electrolyzed reduced water did not improve WBSF or sensory characteristics compared to IS treated chops. As a stand-alone enhancement solution alkaline electrolyzed reduced water was not a suitable replacement for industry standard solutions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Structure of Accelerated Learning Program (ALP) Efforts, 2000-01.

    ERIC Educational Resources Information Center

    Baenen, Nancy; Yaman, Kimberly

    This report focuses on the structure of instructional assistance available through the Accelerated Learning Program (ALP) to students who show low achievement in the Wake County Public School System (WCPSS), North Carolina. Context information is also provided on other programs available to these students. Reports on ALP student participation,…

  9. [How to manage mineral metabolism disorders in renal failure].

    PubMed

    Jean, Guillaume

    2011-11-01

    Mineral metabolism abnormalities are frequently observed in patients with chronic kidney disease (CKD). The bone and cardiovascular consequences should lead to the implementation of some adapted strategies for the prevention and treatment on the basis of the physiopathology of the disease and international recommendations. Biological bone markers such as serum parathyroid hormone (PTH) and alkaline phosphatase (ALP) are necessary to classify bone diseases without the need for bone biopsy. Elevated levels of bone markers are detected in cases of secondary hyperparathyroidism (SHPT), whereas decreased levels are observed in cases of adynamic bone disease (ABD). Bone mineral density, however, is not useful for the diagnosis. Vitamin D supplementation and reducing hyperphosphataemia by dietary phosphate-intake restriction, phosphate binders, and dialysis, are the main steps for the prevention of SHPT. Calcitriol analogs and calcimimetics should be used in second line in cases of SHPT. For the treatment of ABD, excess use of calcium salts and calcitriol analogs need to be avoided. Managing these therapies adequately can help maintain the main biological values (i.e. serum PTH, calcium, phosphorus, and ALP) within their recommended ranges. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  10. Nutrient Limitation Dynamics of a Coastal Cape Cod Pond: Seasonal Trends in Alkaline Phosphatase Activity

    DTIC Science & Technology

    2000-11-13

    Collection and Nutrient Analyses Ashumet Pond water column profiles and samples were taken by the School for Marine Science and Technology (SMAST) at the...Collection & Analysis ........................................ .......... 77 4.3.1 SMAST Water Sampling Plan/Collection and Nutrient Analyses...suited as an indicator of phosphate limitation in natural waters . In this study alkaline phosphatase is used to understand the nutrient limitation

  11. Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration

    PubMed Central

    FENG, ZIHAO; LIU, JIAQI; SHEN, CONGCONG; LU, NANHANG; ZHANG, YONG; YANG, YANWEN; QI, FAZHI

    2016-01-01

    The present study aimed to investigate the properties of a promising bone scaffold for bone repair, which consisted of a novel composite of adipose-derived stem cells (ADSCs) attached to a porous β-tricalcium phosphate (β-TCP) scaffold with platelet-rich plasma (PRP). The β-TCP powder was synthesized and its composition was determined using X-ray diffraction and Fourier transform infrared spectroscopy. The surface morphology and microstructure of the fabricated porous β-TCP scaffold samples were analyzed using light and scanning electron microscopy, and their porosity and compressive strength were also evaluated. In addition, the viability of rabbit ADSCs incubated with various concentrations of the β-TCP extraction fluid was analyzed. The rate of attachment and the morphology of biotinylated ADSCs (Bio-ADSCs) on avidin-coated β-TCP (Avi-β-TCP), and untreated ADSCs on β-TCP, were compared. Furthermore, in vivo bone-forming abilities were determined following the implantation of group 1 (Bio-ADSCs/Avi-β-TCP) and group 2 (Bio-ADSCs/Avi-β-TCP/PRP) constructs using computed tomography, and histological osteocalcin (OCN) and alkaline phosphatase (ALP) expression analyses in a rabbit model of mandibulofacial defects. The β-TCP scaffold exhibited a high porosity (71.26±0.28%), suitable pore size, and good mechanical strength (7.93±0.06 MPa). Following incubation with β-TCP for 72 h, 100% of viable ADSCs remained. The avidin-biotin binding system significantly increased the initial attachment rate of Bio-ADSCs to Avi-β-TCP in the first hour (P<0.01). Following the addition of PRP, group 2 exhibited a bony-union and mandibular body shape, newly formed bone and increased expression levels of OCN and ALP in the mandibulofacial defect area, as compared with group 1 (P<0.05). The results of the present study suggested that the novel Bio-ADSCs/Avi-β-TCP/PRP composite may have potential application in bone repair and bone tissue engineering. PMID:26997987

  12. Crystallization and preliminary X-ray diffraction analysis of a high-affinity phosphate-binding protein endowed with phosphatase activity from Pseudomonas aeruginosa PAO1

    PubMed Central

    Djeghader, Ahmed; Gotthard, Guillaume; Suh, Andrew; Gonzalez, Daniel; Scott, Ken; Chabriere, Eric; Elias, Mikael

    2013-01-01

    In prokaryotes, phosphate starvation induces the expression of numerous phosphate-responsive genes, such as the pst operon including the high-affinity phosphate-binding protein (PBP or pstS) and alkaline phosphatases such as PhoA. This response increases the cellular inorganic phosphate import efficiency. Notably, some Pseudomonas species secrete, via a type-2 secretion system, a phosphate-binding protein dubbed LapA endowed with phosphatase activity. Here, the expression, purification, crystallization and X-ray data collection at 0.87 Å resolution of LapA are described. Combined with biochemical and enzymatic characterization, the structure of this intriguing phosphate-binding protein will help to elucidate the molecular origin of its phosphatase activity and to decipher its putative role in phosphate uptake. PMID:24100568

  13. Liming induces carbon dioxide (CO2) emission in PSB inoculated alkaline soil supplemented with different phosphorus sources.

    PubMed

    Adnan, Muhammad; Shah, Zahir; Sharif, Muhammad; Rahman, Hidayatur

    2018-04-01

    Agricultural land is a major sink of global organic carbon (C). Its suitable management is crucial for improving C sequestration and reducing soil CO 2 emission. Incubation experiments were performed to assess the impact of phosphate solubilizing bacterial (PSB) inoculation (inoculated and uninoculated) and soil calcification (4.78, 10, 15, and 20% crushed CaCO 3 ) with phosphorus (P) sources [single superphosphate (SSP), rock phosphate (RP), farm yard manure (FYM), and poultry manure (PM)] in experiment 1 and with various rates of PM (4, 8, and 12 kg ha -1 ) in experiment 2 on cumulative soil respiration. These experiments were arranged in three factorial, complete randomize design (CRD) with three replications. Interactively, lime with P sources (at day 1 and 3) and lime with PSB (at day 1) significantly expedited soil respiration. Mainly, PSB inoculation, liming, PM fertilization, and its various rates significantly enhanced soil respiration with time over control/minimum in alkaline soil at all incubation periods. Higher CO 2 emission was detected in soil supplemented with organic P sources (PM and FYM) than mineral sources (SSP and RP). CO 2 emission was noted to increase with increasing PM content. Since liming intensified CO 2 discharge from soil, therefore addition of lime to an alkaline soil should be avoided; instead, integrated approaches must be adopted for P management in alkaline calcareous soils for climate-smart agriculture.

  14. Consortium inoculum of five thermo-tolerant phosphate solubilizing Actinomycetes for multipurpose biofertilizer preparation.

    PubMed

    Nandimath, Arusha P; Karad, Dilip D; Gupta, Shantikumar G; Kharat, Arun S

    2017-10-01

    Alkaline pH of the soil facilitates the conversion of phosphate present in phosphate fertilizer applied in the field to insoluble phosphate which is not available to plants. Problem of soluble phosphate deficiency arises, primarily due to needless use of phosphate fertilizer. We sought to biofertilizer with the thermo-tolerant phosphate solubilizing actinomycetes consortium that could convert insoluble phosphate to soluble phosphate at wider temperature range. In the present investigation consortium of five thermo-tolerant phosphate solubilizing actinomycetes was applied for preparation of inoculum to produce multipurpose bio-fertilizer. Phosphates solubilizing thermo-tolerant 32 actinomycetes strains were processed for identification with the use of PIBWIN software and were screened for phosphate solubilizing activity. Amongst these five actinomycetes were selected on the basis of their ability to produce cellulase, chitinase, pectinase, protease, lipase, amylase and phosphate solubilizing enzymes. Ability to produce these enzymes at 28°C and 50°C were examined. Biofertilizer was prepared by using agricultural waste as a raw material. While preparation of bio-fertilizer the pH decreased from 7.5 to 4.3 and temperature increased up to 74°C maximum at the end of 4 th week and in subsequent week it started to decline gradually till it reached around 50°C, which was found to be stable up to eighth week. This thermo-tolerant actinomycetes consortium released soluble phosphate of up to 46.7 μg ml -1 . As the mesophilic organisms die out at high temperature of composting hence thormo-tolerant actinomycetes would be the better substitute for preparation of phosphate solubilizing bio-fertilizer with added potential to degrade complex macromolecules in composting.

  15. Circadian and longitudinal variation of serum C-telopeptide, osteocalcin, and skeletal alkaline phosphatase in C3H/HeJ mice.

    PubMed

    Srivastava, A K; Bhattacharyya, S; Li, X; Mohan, S; Baylink, D J

    2001-10-01

    Inbred strains of mice are increasingly being used as an animal model to investigate skeletal disorders relevant to humans. In the bone field, one of the most convenient endpoints for evaluating genetic, physiological, or pharmaceutical perturbations is the use of biochemical markers. To apply biochemical markers in an effective manner, it is of key importance to establish the biological variation and appropriate sampling time. In this study, we evaluate two components: (i) circadian changes, and (ii) longitudinal variation for three serum markers, osteocalcin, C-telopeptide, and skeletal alkaline phosphatase (sALP), using 6-week-old C3H/HeJ (C3H) mice. To study circadian rhythms, the mice were randomly divided into eight groups of 15 mice each. Blood was collected at 3 h intervals, starting at 9:00 A.M. and continuing until 6:00 A.M. the next day. To determine whether circadian rhythm is intrinsically regulated or influenced by restricted food intake, it was also studied after a 12 h fasting period. Serum osteocalcin and C-telopeptide levels were measured by enzyme-linked immunoassay (ELISA) and skeletal alkaline phosphatase by a kinetic assay. The results demonstrated significant circadian variations in osteocalcin and C-telopeptide levels with a peak value between 0900 and 1200 h during daytime and a nadir between 15:00 and 18:00 h. The peak levels of C-telopeptide and osteocalcin were 26%-66% higher as compared with 24 h mean values. The pattern of the circadian variation of C-telopeptide and osteocalcin was similar in female and male animals and was not significantly affected by restricted food intake. The sALP levels were only marginally affected by the circadian rhythm. Longitudinal variations, expressed as coefficient of variation (CV), for osteocalcin, C-telopeptide, and sALP concentrations were 17%, 14%, and 16%, respectively. In addition, the longitudinal variations were not significantly influenced by the time of blood collection in sALP and osteocalcin

  16. A nanohybrid of platinum nanoparticles-porous ZnO-hemin with electrocatalytic activity to construct an amplified immunosensor for detection of influenza.

    PubMed

    Yang, Zhe-Han; Zhuo, Ying; Yuan, Ruo; Chai, Ya-Qin

    2016-04-15

    In this work, a nanohybrid of platinum nanoparticles-porous ZnO spheres-hemin (Pt-pZnO-hemin) was synthesized for construction of alkaline phosphatase-based immunosensor for detection of influenza. Briefly, porous ZnO spheres (pZnO) were prepared using soluble starches as the capping agent, followed by surface functionalization of platinum nanoparticles via a hydrothermal method (Pt-pZnO). Then, hemin with carboxylic functionality was spontaneously adsorbed onto Pt-pZnO by ester-like binding between carboxylic group of hemin and ZnO. Compared with platinum nanoparticles and hemin, the resulting Pt-pZnO-hemin nanohybrid showed more excellent electrocatalysis activity toward 1-naphthol (1-NP). Taking advantage of the Pt-pZnO-hemin, we have developed an amplified electrochemical immunosensor based on in situ generation of redox probe by alkaline phosphatase (ALP) and Pt-pZnO-hemin as signal enhancer. Herein, electrochemically active 1-NP was generated by enzymatic hydrolysis of inactive 1-naphthyl phosphate by ALP, then Pt-pZnO-hemin was used as catalyst to catalytically oxidize 1-NP, resulting in electrochemical signal amplification. Furthermore, in comparison with other nanomaterials including Au-pZnO, Pt-pZnO and Au-pZnO-hemin, the excellent catalytical property of Pt-pZnO-hemin make it a promising nanohybrid material for ALP-based immunosensor for signal amplification. Copyright © 2015. Published by Elsevier B.V.

  17. Low-intensity pulsed ultrasound (LIPUS) stimulates mineralization of MC3T3-E1 cells through calcium and phosphate uptake.

    PubMed

    Tassinary, João Alberto Fioravante; Lunardelli, Adroaldo; Basso, Bruno de Souza; Dias, Henrique Bregolin; Catarina, Anderson Velasque; Stülp, Simone; Haute, Gabriela Viegas; Martha, Bianca Andrade; Melo, Denizar Alberto da Silva; Nunes, Fernanda Bordignon; Donadio, Márcio Vinícius Fagundes; Oliveira, Jarbas Rodrigues de

    2018-03-01

    The present study aimed to evaluate the effect of low-intensity pulsed ultrasound (LIPUS) on pre-osteoblast mineralization using in vitro bioassays. Pre-osteoblastic MC3T3-E1 cells were exposed to LIPUS at 1 MHz frequency, 0.2 W/cm 2 intensity and 20% duty cycle for 30 min. The analyses were carried out up to 336 h (14 days) after exposure. The concentration of collagen, phosphate, alkaline phosphatase, calcium and transforming growth factor beta 1 (TGF-β1) in cell supernatant and the presence of calcium deposits in the cells were analyzed. Our results showed that LIPUS promotes mineralized nodules formation. Collagen, phosphate, and calcium levels were decreased in cell supernatant at 192 h after LIPUS exposure. However, alkaline phosphatase and TGF-β1 concentrations remained unchanged. Therapeutic pulsed ultrasound is capable of stimulating differentiation and mineralization of pre-osteoblastic MC3T3-E1 cells by calcium and phosphate uptake with consequent hydroxyapatite formation. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Comparison of osteoblast-like cell responses to calcium silicate and tricalcium phosphate ceramics in vitro.

    PubMed

    Ni, Siyu; Chang, Jiang; Chou, Lee; Zhai, Wanyin

    2007-01-01

    Calcium silicate ceramics have been proposed as new bone repair biomaterials, since they have proved to be bioactive, degradable, and biocompatible. Beta-tricalcium phosphate ceramic is a well-known degradable material for bone repair. This study compared the effects of CaSiO3 (alpha-, and beta-CaSiO3) and beta-Ca3(PO4)2 (beta-TCP) ceramics on the early stages of rat osteoblast-like cell attachment, proliferation, and differentiation. Osteoblast-like cells were cultured directly on CaSiO3 (alpha-, and beta-CaSiO3) and beta-TCP ceramics. Attachment of a greater number of cells was observed on CaSiO3 (alpha-, and beta-CaSiO3) ceramics compared with beta-TCP ceramics after incubation for 6 h. SEM observations showed an intimate contact between cells and the substrates, significant cells adhesion, and that the cells spread and grew on the surfaces of all the materials. In addition, the proliferation rate and alkaline phosphatase (ALP) activity of the cells on the CaSiO3 (alpha-, and beta-CaSiO3) ceramics were improved when compared with the beta-TCP ceramics. In the presence of CaSiO3, elevated levels of calcium and silicon in the culture medium were observed throughout the 7-day culture period. In conclusion, the results of the present study revealed that CaSiO3 ceramics showed greater ability to support cell attachment, proliferation, and differentiation than beta-TCP ceramic. 2006 Wiley Periodicals, Inc.

  19. Amorphous cobalt potassium phosphate microclusters as efficient photoelectrochemical water oxidation catalyst

    NASA Astrophysics Data System (ADS)

    Zhang, Ye; Zhao, Chunsong; Dai, Xuezeng; Lin, Hong; Cui, Bai; Li, Jianbao

    2013-12-01

    A novel amorphous cobalt potassium phosphate hydrate compound (KCoPO4·H2O) is identified to be active photocatalyst for oxygen evolution reaction (OER) to facilitate hydrogen generation from water photolysis. It has been synthesized through a facile and cost-effective solution-based precipitation method using earth-abundant materials. Its highly porous structure and large surface areas are found to be responsible for the excellent electrochemical performance featuring a low OER onset at ∼550 mVSCE and high current density in alkaline condition. Unlike traditional cobalt-based spinel oxides (Co3O4, NiCo2O4) and phosphate (Co-Pi, Co(PO3)2) electrocatalysts, with proper energy band alignment for light-assisted water oxidation, cobalt potassium phosphate hydrate also exhibits robust visible-light response, generating a photocurrent density of ∼200 μA cm-2 at 0.7 VSCE. This catalyst could thus be considered as a promising candidate to perform photoelectrochemical water splitting.

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

    NASA Astrophysics Data System (ADS)

    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

  1. Evaluation of pink-pigmented facultative methylotrophic bacteria for phosphate solubilization.

    PubMed

    Jayashree, Shanmugam; Vadivukkarasi, Ponnusamy; Anand, Kirupanithi; Kato, Yuko; Seshadri, Sundaram

    2011-08-01

    Thirteen pink-pigmented facultative methylotrophic (PPFM) strains isolated from Adyar and Cooum rivers in Chennai and forest soil samples in Tamil Nadu, India, along with Methylobacterium extorquens, M. organophilum, M. gregans, and M. komagatae were screened for phosphate solubilization in plates. P-solubilization index of the PPFMs grown on NBRIP-BPB plates for 7 days ranged from 1.1 to 2.7. The growth of PPFMs in tricalcium phosphate amended media was found directly proportional to the glucose concentration. Higher phosphate solubilization was observed in four strains MSF 32 (415 mg l(-l)), MDW 80 (301 mg l(-l)), M. komagatae (279 mg l(-l)), and MSF 34 (202 mg l(-l)), after 7 days of incubation. A drop in the media pH from 6.6 to 3.4 was associated with an increase in titratable acidity. Acid phosphatase activity was more pronounced in the culture filtrate than alkaline phosphatase activity. Adherence of phosphate to densely grown bacterial surface was observed under scanning electron microscope after 7-day-old cultures. Biochemical characterization and screening for methanol dehydrogenase gene (mxaF) confirmed the strains as methylotrophs. The mxaF gene sequence from MSF 32 clustered towards M. lusitanum sp. with 99% similarity. This study forms the first detailed report on phosphate solubilization by the PPFMs.

  2. Nuclear translocation of the cytoskeleton-associated protein, smALP, upon induction of skeletal muscle differentiation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cambier, Linda; Pomies, Pascal, E-mail: pascal.pomies@crbm.cnrs.fr

    2011-06-17

    Highlights: {yields} The cytoskeleton-associated protein, smALP, is expressed in differentiated skeletal muscle. {yields} smALP is translocated from the cytoplasm to the nucleus of C2C12 myoblasts upon induction of myogenesis. {yields} The differentiation-dependent nuclear translocation of smALP occurs in parallel with the nuclear accumulation of myogenin. {yields} The LIM domain of smALP is essential for the nuclear accumulation of the protein. {yields} smALP might act in the nucleus to control some critical aspect of the muscle differentiation process. -- Abstract: The skALP isoform has been shown to play a critical role in actin organization and anchorage within the Z-discs of skeletalmore » muscles, but no data is available on the function of the smALP isoform in skeletal muscle cells. Here, we show that upon induction of differentiation a nuclear translocation of smALP from the cytoplasm to the nucleus of C2C12 myoblasts, concomitant to an up-regulation of the protein expression, occurs in parallel with the nuclear accumulation of myogenin. Moreover, we demonstrate that the LIM domain of smALP is essential for the nuclear translocation of the protein.« less

  3. The effect of phosphomonoesterases on the oxygen isotope composition of phosphate

    NASA Astrophysics Data System (ADS)

    von Sperber, Christian; Kries, Hajo; Tamburini, Federica; Bernasconi, Stefano M.; Frossard, Emmanuel

    2014-01-01

    Plants and microorganisms under phosphorus (P) stress release extracellular phosphatases as a strategy to acquire inorganic phosphate (Pi). These enzymes catalyze the hydrolysis of phosphoesters leading to a release of Pi. During the enzymatic hydrolysis an isotopic fractionation (ε) occurs leaving an imprint on the oxygen isotope composition of the released Pi which might be used to trace phosphorus in the environment. Therefore, enzymatic assays with acid phosphatases from wheat germ and potato tuber and alkaline phosphatase from Escherichia coli were prepared in order to determine the oxygen isotope fractionation caused by these enzymes. Adenosine 5‧ monophosphate and glycerol phosphate were used as substrates. The oxygen isotope fractionation caused by acid phosphatases is 20-30‰ smaller than for alkaline phosphatases, resulting in a difference of 5-7.5‰ in δ18O of Pi depending on the enzyme. We attribute the enzyme dependence of the isotopic fractionation to distinct reaction mechanisms of the two types of phosphatases. The observed difference is large enough to distinguish between the two enzymatic processes in environmental samples. These findings show that the oxygen isotope composition of Pi can be used to trace different enzymatic processes, offering an analytical tool that might contribute to a better understanding of the P-cycle in the environment.

  4. Separating and stabilizing phosphate from high-level radioactive waste: process development and spectroscopic monitoring.

    PubMed

    Lumetta, Gregg J; Braley, Jenifer C; Peterson, James M; Bryan, Samuel A; Levitskaia, Tatiana G

    2012-06-05

    Removing phosphate from alkaline high-level waste sludges at the Department of Energy's Hanford Site in Washington State is necessary to increase the waste loading in the borosilicate glass waste form that will be used to immobilize the highly radioactive fraction of these wastes. We are developing a process which first leaches phosphate from the high-level waste solids with aqueous sodium hydroxide, and then isolates the phosphate by precipitation with calcium oxide. Tests with actual tank waste confirmed that this process is an effective method of phosphate removal from the sludge and offers an additional option for managing the phosphorus in the Hanford tank waste solids. The presence of vibrationally active species, such as nitrate and phosphate ions, in the tank waste processing streams makes the phosphate removal process an ideal candidate for monitoring by Raman or infrared spectroscopic means. As a proof-of-principle demonstration, Raman and Fourier transform infrared (FTIR) spectra were acquired for all phases during a test of the process with actual tank waste. Quantitative determination of phosphate, nitrate, and sulfate in the liquid phases was achieved by Raman spectroscopy, demonstrating the applicability of Raman spectroscopy for the monitoring of these species in the tank waste process streams.

  5. Safety and Efficacy of Stosstherapy in Nutritional Rickets.

    PubMed

    Chatterjee, Daipayan; Swamy, Mathad K S; Gupta, Vikas; Sharma, Vasu; Sharma, Akshat; Chatterjee, Krishti

    2017-03-01

    Stosstherapy has been used since early 19 th century for treating nutritional rickets. However, there are no clear cut guidelines for the biochemical monitoring of this treatment. Repeated blood tests at short intervals increase the cost of therapy and noncompliance. A prospective study was conducted on 191 cases of nutritional rickets below 10 years of age to evaluate the effectivity of stosstherapy. All cases were treated with a single intramuscular injection of vitamin D (600.000 IU) along with oral calcium (50 mg/kg) and vitamin D (400 IU per day) until radiological resolution. Dietary modifications and adequate sunlight exposure were also recommended. The mean age of presentation was 2 years 9 months. Mean sunlight exposure was 17 minutes/week with 90% having low sunlight exposure (<30 minutes/week). Prolonged breast feeding (>6 months) was found in 93.7% of the cases. With treatment, the clinical features started resolving by 1 month with complete resolution of most of the features over a period of 1 year. By 6 months, all the study subjects had complete radiological resolution. Serum levels of calcium and alkaline phosphatase (ALP) were restored by 6 months in most cases while phosphate and vitamin D levels normalized by 6 weeks. Stosstherapy is a safe, cheap and effective method of treating nutritional rickets. Biochemical tests at initial presentation followed by vitamin D assay at 6 weeks and calcium, phosphate and ALP assays at 6 months is recommended in the monitoring of these patients. For regular monitoring, only ALP assay is recommended, provided one abstains from repeat injection of vitamin D based on high ALP levels.

  6. Algorithms used in the Airborne Lidar Processing System (ALPS)

    USGS Publications Warehouse

    Nagle, David B.; Wright, C. Wayne

    2016-05-23

    The Airborne Lidar Processing System (ALPS) analyzes Experimental Advanced Airborne Research Lidar (EAARL) data—digitized laser-return waveforms, position, and attitude data—to derive point clouds of target surfaces. A full-waveform airborne lidar system, the EAARL seamlessly and simultaneously collects mixed environment data, including submerged, sub-aerial bare earth, and vegetation-covered topographies.ALPS uses three waveform target-detection algorithms to determine target positions within a given waveform: centroid analysis, leading edge detection, and bottom detection using water-column backscatter modeling. The centroid analysis algorithm detects opaque hard surfaces. The leading edge algorithm detects topography beneath vegetation and shallow, submerged topography. The bottom detection algorithm uses water-column backscatter modeling for deeper submerged topography in turbid water.The report describes slant range calculations and explains how ALPS uses laser range and orientation measurements to project measurement points into the Universal Transverse Mercator coordinate system. Parameters used for coordinate transformations in ALPS are described, as are Interactive Data Language-based methods for gridding EAARL point cloud data to derive digital elevation models. Noise reduction in point clouds through use of a random consensus filter is explained, and detailed pseudocode, mathematical equations, and Yorick source code accompany the report.

  7. Consortium inoculum of five thermo-tolerant phosphate solubilizing Actinomycetes for multipurpose biofertilizer preparation

    PubMed Central

    Nandimath, Arusha P.; Karad, Dilip D.; Gupta, Shantikumar G.; Kharat, Arun S.

    2017-01-01

    Background and Objectives: Alkaline pH of the soil facilitates the conversion of phosphate present in phosphate fertilizer applied in the field to insoluble phosphate which is not available to plants. Problem of soluble phosphate deficiency arises, primarily due to needless use of phosphate fertilizer. We sought to biofertilizer with the thermo-tolerant phosphate solubilizing actinomycetes consortium that could convert insoluble phosphate to soluble phosphate at wider temperature range. Materials and Methods: In the present investigation consortium of five thermo-tolerant phosphate solubilizing actinomycetes was applied for preparation of inoculum to produce multipurpose bio-fertilizer. Phosphates solubilizing thermo-tolerant 32 actinomycetes strains were processed for identification with the use of PIBWIN software and were screened for phosphate solubilizing activity. Results: Amongst these five actinomycetes were selected on the basis of their ability to produce cellulase, chitinase, pectinase, protease, lipase, amylase and phosphate solubilizing enzymes. Ability to produce these enzymes at 28°C and 50°C were examined. Biofertilizer was prepared by using agricultural waste as a raw material. While preparation of bio-fertilizer the pH decreased from 7.5 to 4.3 and temperature increased up to 74°C maximum at the end of 4th week and in subsequent week it started to decline gradually till it reached around 50°C, which was found to be stable up to eighth week. This thermo-tolerant actinomycetes consortium released soluble phosphate of up to 46.7 μg ml−1. Conclusion: As the mesophilic organisms die out at high temperature of composting hence thormo-tolerant actinomycetes would be the better substitute for preparation of phosphate solubilizing bio-fertilizer with added potential to degrade complex macromolecules in composting. PMID:29296275

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

    PubMed

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

    2015-02-03

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

  9. Si3 AlP: A New Promising Material for Solar Cell Absorber

    NASA Astrophysics Data System (ADS)

    Yang, Jihui; Zhai, Yingteng; Liu, Hengrui; Xiang, Hongjun; Gong, Xingao; Wei, Suhuai

    2014-03-01

    First-principles calculations are performed to study the structural and optoelectronic properties of the newly synthesized nonisovalent and lattice-matched (Si2)0.6(AlP)0.4 alloy [T. Watkins et al., J. Am. Chem. Soc. 2011, 133, 16212.] The most stable structure of Si3AlP is a superlattice along the <111>direction with separated AlP and Si layers, which has a similar optical absorption spectrum to silicon. The ordered C1c1-Si3AlP is found to be the most stable one among all the structures with -AlPSi3- motifs, in agreement with the experimental suggestions. We predict that C1c1-Si3AlP has good optical properties, i.e., it has a larger fundamental band gap and a smaller direct band gap than Si, thus it has much higher absorption in the visible light region, making it a promising candidate for improving the performance of the existing Si-based solar cells.

  10. Younger Dryas equilibrium line altitudes and precipitation patterns in the Alps

    NASA Astrophysics Data System (ADS)

    Kerschner, Hanns; Moran, Andrew; Ivy-Ochs, Susan

    2016-04-01

    Moraine systems of the "Egesen Stadial" are widespread and easily identifiable features in the Alps. Absolute dating with terrestrial cosmogenic radionuclides shows that the maximum extent was reached during the early Younger Dryas (YD), probably as a reaction to the intense climatic downturn subsequent to Lateglacial Interstadial. In recent years, several new studies and the availability of high-quality laser-scan hillshades and orthophotos allowed a significant extension of the database of YD glaciers as "palaeoprecipitation gauges" to large hitherto unmapped regions in the Austrian and Swiss Alps. The equilibrium line altitude (ELA) of the glaciers and its lowering relative to the Little Ice Age ELA (dELA) shows a distinct and systematic spatial pattern. Along the northern slope of the Alps, dELAs are usually large (around 400 m and perhaps even more), while dELAs range around 200 m in the well sheltered areas of the central Alps, e.g. in the Engadine and in western Tyrol. Both stochastic glacier-climate models (e.g. Ohmura et al. 1992) and the heat- and mass balance equation (Kuhn 1981) allow the reconstruction of precipitation change under the assumption of a spatially constant summer temperature depression, which in turn can be estimated from biological proxies. This allows to draw the spatial pattern of precipitation change with considerable detail. Precipitation change is clearly controlled by the local relief like high mountain chains, deeply incised and long valleys and mountain passes. Generally the contrast between the northern fringe of the Alps and the interior was more pronounced than today. Climate in the Northern and and Northwestern Alps was rather wet with precipitation totals eventually exceeding modern annual sums. The central Alps received 20 - 30% less precipitation than today, mainly due to reduced winter precipitation. In the southern Alps, still scarce spatial information points to precipitation sums which were approximately similar to

  11. Phenology in the Western Alps: first results of the PhenoALP project

    NASA Astrophysics Data System (ADS)

    Cremonese, Edoardo; Tracol, Yann

    2010-05-01

    PHENOALP is a EU co-funded Interreg Project under the operational programme for cross-border cooperation "Italy-France (Alps-ALCOTRA)" 2007 - 2013, started in 2009, aiming to get a better understanding of phenological changes in the Alps and build a long term monitoring network. The results obtained after the first year of the project are mainly related to the definition of observation protocols and to the implementation of the observation networks. In particular, we focused on the comparison of different approaches for monitoring alpine grasslands phenology. We developed a new protocol for vegetative and reproductive phases of the seven most common plant growth life forms of alpine pastures: cyperaceae, poaceae (palatable and non palatable), evergreen and deciduous shrubs, forbs and leguminous. For each group quantitative and qualitative variables (e.g. leaves length, bud number, fruits number and phenophases) are monitored during the growing season. Study sites are located along an elevation gradient from 1560 to 2580 m asl and measurements are carried out on marked individuals in permanent plots. The other techniques used to monitor grassland phenology are: analysis of webcam images, weekly nadiral digital images, visual estimations of greening percentage, canopy structural measurements (i.e. height, fraction of absorbed photosynthetically active radiation, leaf are index, etc..) and high frequency radiometric measurements of vegetation indexes related to canopy structure. All methods are providing promising results and our goal is to define a protocol suitable for long term observation based on a reasonable trade-off between the quality and robustness of collected data and the heaviness of the observations. For animal phenology we are focusing on many animal taxa among birds, mammals, amphibians and insects. First results are coming from birds and amphibians. In the case of birds, observations of reproductive phenology of some common alpine species are done

  12. The Grand St Bernard-Briançonnais Nappe System and the Paleozoic Inheritance of the Western Alps Unraveled by Zircon U-Pb Dating

    NASA Astrophysics Data System (ADS)

    Bergomi, M. A.; Dal Piaz, G. V.; Malusà, M. G.; Monopoli, B.; Tunesi, A.

    2017-12-01

    The continental crust involved in the Alpine orogeny was largely shaped by Paleozoic tectono-metamorphic and igneous events during oblique collision between Gondwana and Laurussia. In order to shed light on the pre-Alpine basement puzzle disrupted and reamalgamated during the Tethyan rifting and the Alpine orogeny, we provide sensitive high-resolution ion microprobe U-Pb zircon and geochemical whole rock data from selected basement units of the Grand St Bernard-Briançonnais nappe system in the Western Alps and from the Penninic and Lower Austroalpine units in the Central Alps. Zircon U-Pb ages, ranging from 459.0 ± 2.3 Ma to 279.1 ± 1.1 Ma, provide evidence of a complex evolution along the northern margin of Gondwana including Ordovician transtension, Devonian subduction, and Carboniferous-to-Permian tectonic reorganization. Original zircon U-Pb ages of 371 ± 0.9 Ma and 369.3 ± 1.5 Ma, from calc-alkaline granitoids of the Grand Nomenon and Gneiss del Monte Canale units, provide the first compelling evidence of Late Devonian orogenic magmatism in the Alps. We propose that rocks belonging to these units were originally part of the Moldanubian domain and were displaced toward the SW by Late Carboniferous strike-slip faulting. The resulting assemblage of basement units was disrupted by Permian tectonics and by Mesozoic opening of the Alpine Tethys. Remnants of the Moldanubian domain became either part of the European paleomargin (Grand Nomenon unit) or part of the Adriatic paleomargin (Gneiss del Monte Canale unit), to be finally accreted into the Alpine orogenic wedge during the Cenozoic.

  13. Overestimation of organic phosphorus in wetland soils by alkaline extraction and molybdate colorimetry.

    PubMed

    Turner, Benjamin L; Newman, Susan; Reddy, K Ramesh

    2006-05-15

    Accurate information on the chemical nature of soil phosphorus is essential for understanding its bioavailability and fate in wetland ecosystems. Solution phosphorus-31 nuclear magnetic resonance (31P NMR) spectroscopy was used to assess the conventional colorimetric procedure for phosphorus speciation in alkaline extracts of organic soils from the Florida Everglades. Molybdate colorimetry markedly overestimated organic phosphorus by between 30 and 54% compared to NMR spectroscopy. This was due in large part to the association of inorganic phosphate with organic matter, although the error was exacerbated in some samples by the presence of pyrophosphate, an inorganic polyphosphate that is not detected by colorimetry. The results have important implications for our understanding of phosphorus biogeochemistry in wetlands and suggest that alkaline extraction and solution 31p NMR spectroscopy is the only accurate method for quantifying organic phosphorus in wetland soils.

  14. Synthesis and evaluation of benzo[b]thiophene derivatives as inhibitors of alkaline phosphatases.

    PubMed

    Li, Lina; Chang, Lei; Pellet-Rostaing, Stéphane; Liger, François; Lemaire, Marc; Buchet, René; Wu, Yuqing

    2009-10-15

    Presence of basic calcium phosphate in knee joints of osteoarthritis patients could be prevented by inhibiting tissue non-specific alkaline phosphatase (TNAP) activity. Levamisole or the L stereoisomer of tetramisole (a known TNAP inhibitor) has been used as a treatment for curing rheumatoid arthritis but its therapeutical use is limited due to side effects. We report the synthesis and the TNAP inhibition property of benzo[b]thiophene derivatives, among which benzothiopheno-tetramisole and benzothiopheno-2,3-dehydrotetramisole, which could be involved in a drug therapy for osteoarthritis. Two water soluble racemic benzothiopheno-tetramisole and -2,3-dehydrotetramisole with apparent inhibition constants K(i)=85+/-6 microM and 135+/-3 microM (n=3) comparable to that of enantiomeric levamisole 93+/-4 microM were found. Several novel derivatives showed more pronounced inhibition properties towards intestinal alkaline phosphatase than TNAP.

  15. Low temperature thermochronology in the Easter Alps. New data, interpretations and perspectives.

    NASA Astrophysics Data System (ADS)

    Wölfler, Andreas

    2015-04-01

    The aim of this study is to evaluate new and published low temperature thermochronological data of the Eastern Alps in terms of its Mesozoic and Cenozoic tectonic evolution and the possible connection with deep seated lithospheric processes. In the Eastern Alps, the tectonic units that originate from the Penninic domain are buried beneath the Austroalpine nappe stack. Overthrusting of the Austroalpine nappes over the Penninic units occurred throughout the Cretaceous and lasted until the Eocene. During lateral tectonic extrusion in Oligocene to Miocene times the footwall penninic units exposed in the Tauern Window, were tectonically exhumed from below the Austroalpine hanging wall. This is well documented by Miocene to Pliocene zircon- and apatite fission track (ZFT, AFT) and (U-Th)/He data. However, the Austroalpine hanging wall shows a more complex age pattern. Late Cretaceous ZFT data reflect post-metamorphic exhumational cooling after Eo-Alpine metamorphism that goes along with an extensional phase that affected large parts of the Eastern Alps. Paleogene AFT and apatite (U-Th)/He data of the Austroalpine units to the east of the Tauern Window reflect exhumation of this area that supplied clastic material, the so-called Augenstein formation. Exhumation and erosion of the area left a probably hilly surface in Early Miocene times that was only moderately uplifted since then. These areas are well known for paleosurfaces exposed in the Gurktal- Kor- and Seckauer Alps to the east of the Tauern Window and in the central and eastern Northern Calcareous Alps. However, distinct parts of the Austroalpine hanging wall experienced substantial exhumation and surface uplift in the Miocene, contemporaneous to the exhumation of Penninic units and lateral extrusion of the Eastern Alps. These areas are restricted to the south and northeast of the Tauern Window and are characterized by steep and rugged reliefs that contrast the hilly and moderately shaped reliefs of the

  16. Seismotectonics investigations in the internal Cottian Alps (Italian Western Alps)

    NASA Astrophysics Data System (ADS)

    Perrone, Gianluigi; Eva, Elena; Solarino, Stefano; Cadoppi, Paola; Balestro, Gianni; Fioraso, Gianfranco; Tallone, Sergio

    2010-05-01

    The inner Cottian Alps represent an area of a low- to moderate- magnitude seismicity (Eva et al., 1990) even though some historical earthquakes reached VIII degree of the Mercalli's scale. Although the frame of seismicity is quite well known, the relation between faults and earthquake sources is still under debate. The low deformation rates and the occurrence of several glacial-interglacial cycles during the Pleistocene partly masked the geomorphological evidences of the recent tectonic activity. Recent studies based on field mapping and structural analysis (Balestro et al., 2009; Perrone et al., 2009) allowed characterizing the size and extension of the regional-scale faults dissecting this area of the Western Alps. Here, we combine the results of these novel studies and updated seismological data with the aim to investigate the relations between mapped faults and seismic activity. In the analyzed area both continental crust and oceanic tectonic units, belonging to the Penninic Domain of the Western Alps, crop out. The main brittle tectonic feature of this area is represented by the Lis-Trana Deformation Zone (LTZ), an N-S striking, steep structure that extends for about 35 km from the Lower Lanzo valleys to the Lower Sangone Valley. The occurrence of steep faults displacing the metamorphic basement, showed in seismic sections carried out for oil exploration (Bertotti & Mosca, 2009), suggests that the LTZ may be prolonged Southward beneath the Plio-Quaternary deposits of the Po Plain. West of the LTZ some other minor E-W and N-S faults are also present. Zircon and apatite fission-track data indicate that the activity of these faults started since the Oligocene. Two main faulting stages characterize the post-metamorphic structural evolution of this area: the earlier (faulting stage A; Oligocene?-Early Miocene?) is associated to right-lateral movements along the LTZ and sinistral movements along E-W faults; the subsequent faulting stage (faulting stage B; post

  17. M13 phage peptide ZL4 exerts its targeted binding effect on schistosoma japonicum via alkaline phosphatase.

    PubMed

    Liu, Yan; Yang, Shenghui; Xiao, Jianhua; Yu, Liang; Chen, Li; Zou, Ju; Wang, Kegeng; Tan, Sijie; Yu, Zhengyang; Zeng, Qingren

    2015-01-01

    The present study was to determine the targeting effect of M13 phage peptide ZL4 (MppZL4) on Schistosoma japonicum (S.j). Mice infected with S.j were injected with MppZL4. Real-time PCR was used to detect the distribution and metabolism of MppZL4 in the livers and lungs of mice. In vivo refusion test was performed to detect the targeting of MppZL4. Western blotting was employed to determine the expression of MppZL4. Live imaging was used to detect the distribution of oligopeptide MppZL4. Immunohistochemistry was employed to determine MppZL4 location on adult S.j body surface. Gomori method was employed to detect the influence of oligopeptide MppZL4 on alkaline phosphatase activity. The distribution and metabolism of MppZL4 and M13KE are not significantly different from each other at each time point. The abundance of MppZL4 is changed as S.j migrates in mice. The targeted binding effect of MppZL4 varies at different stages. ZL4 oligopeptide targets S.j in mice. The specific binding sites of MppZL4 on S.j body are mainly located in syncytial cells. The binding sites of MppZL4 on S.j body surface might be ALP or ALP-related proteins. MppZL4 had targeted binding effect on S.j with its binding site being associated with proteins related to S.j alkaline phosphatase. S.j tegument had a specifically binding site with exogenous peptides, offering new means to explore the interactions between hosts and parasites. Additionally, MppZL4 can possibly be used as targeting molecules in worm-resistant drugs or as tracing molecules in imaging diagnosis technologies.

  18. Effect of controlled zinc release on bone mineral density from injectable Zn-containing beta-tricalcium phosphate suspension in zinc-deficient diseased rats.

    PubMed

    Otsuka, Makoto; Ohshita, Yuko; Marunaka, Sunao; Matsuda, Yoshihia; Ito, Atsuo; Ichinose, Noboru; Otsuka, Kuniko; Higuchi, William I

    2004-06-01

    The purpose of this study was to evaluate the efficacy of zinc (Zn)-containing beta-tricalcium phosphate (Zn-TCP) in correcting the bone mineral deficiency noted in osteoporosis using ovariectomized rat model. Four rats were used for each of the four experimental groups: D0, D10, D20, and N10. The rats in D0, D10, and D20 groups were ovariectomized, and fed a vitamin D-, Ca-, and Zn-deficient diet, and induced Zn-deficient osteoporoses for 9 weeks. In contrast, the N10 group was the normal rats fed normal healthy diet for 9 weeks. D0 group was injected with pure beta-TCP suspension, D10 and D20 groups were injected with suspensions containing 10 mg of 10 mol % (6.17 wt % Zn) and 20 mol % (12.05 wt % Zn) Zn-TCP, respectively, and the healthy group, N10 were injected with 10 mol %. Zn-TCP suspensions. Injections were administered intramuscularly in the left thigh once a week in all rats, and fed a vitamin D- and Zn-deficient diet for 9 weeks. The plasma calcium (Ca) and Zn levels, plasma alkaline phosphatase activity (ALP) and bone mineral density (BMD) of the lumbar vertebra and femora were measured. The plasma Zn levels in all the rats were between 1.1 and 2.8 microg/mL. The areas under the curves for the Ca, Zn, and ALP (Ca-AUC, Zn-AUC, and ALP-AUC) levels between 0 and 63 days were calculated. Results for the AUCs were as follows: (1) the Zn-AUCs were in the order of N10 = D20 > D10 > D0; (2) the Ca-AUCs for D0, D10 groups were significantly lower than that for the N10 group; (3) the ALP-AUCs for the D10 and D20 groups were significantly higher than that for the N10 group, and that of the D0 group was in between those. The body weight of D10 and D20 groups significantly increased with time, that of the D0 group increased slightly, and that of the N10 group remained unchanged for the entire experimental period. The BMD of the lumbar vertebrae of the D10 and D20 groups (about 100 mg/cm(2)) was significantly higher than that of the D0 group but lower than that of

  19. Tricyclic coumarin sulphonate derivatives with alkaline phosphatase inhibitory effects: in vitro and docking studies.

    PubMed

    Iqbal, Jamshed; El-Gamal, Mohammed I; Ejaz, Syeda Abida; Lecka, Joanna; Sévigny, Jean; Oh, Chang-Hyun

    2018-12-01

    Tissue-nonspecific alkaline phosphatase (TNAP) is an important isozyme of alkaline phosphatases, which plays different pivotal roles within the human body. Most importantly, it is responsible for maintaining the balanced ratio of phosphate and inorganic pyrophosphate, thus regulates the extracellular matrix calcification during bone formation and growth. The elevated level of TNAP has been linked to vascular calcification and end-stage renal diseases. Consequently, there is a need to search for highly potent and selective inhibitors of alkaline phosphatases (APs) for treatment of disorders associated with the over-expression of APs. Herein, a series of tricyclic coumarin sulphonate 1a-za with known antiproliferative activity, was evaluated for AP inhibition against human tissue nonspecific alkaline phosphatase (h-TNAP) and human intestinal alkaline phosphatase (h-IAP). The methylbenzenesulphonate derivative 1f (IC 50  = 0.38 ± 0.01 μM) was found to be the most active h-TNAP inhibitor. Another 4-fluorobenzenesulphonate derivative 1i (IC 50  = 0.45 ± 0.02 μM) was found as the strongest inhibitor of h-IAP. Some of the derivatives were also identified as highly selective inhibitors of APs. Detailed structure-activity relationship (SAR) was investigated to identify the functional groups responsible for the effective inhibition of AP isozymes. The study was also supported by the docking studies to rationalise the most possible binding site interactions of the identified inhibitors with the targeted enzymes.

  20. New Zealand's Southern Alps

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The rugged Southern Alps extend some 650 kilometers along the western side of New Zealand's South Island. The mountains are often obscured by clouds, which is probably why the Maoris called New Zealand 'Aotearoa', the long white cloud. The higher peaks are snow-covered all year round. Westerly winds bring clouds that drop over 500 centimeters of rain annually on luxuriant rain forest along the west coast. The drier eastern seaboard is home to the majority of the island's population.

    This pair of MISR images is from April 13, 2000 (Terra orbit 1712). The upper image is a natural color view from the instrument's vertical-viewing (nadir) camera. It is presented at a resolution of 550 meters per pixel. The lower image is a stereo anaglyph generated from the instrument's 46-degree and 26-degree forward-viewing cameras, and is presented at 275-meter per pixel resolution to show the portion of the image containing the Southern Alps in greater detail. Viewing the anaglyph in 3-D requires the use of red/blue glasses with the red filter over your left eye. To facilitate stereoscopic viewing, both images have been oriented with north at the left.

    The tallest mountain in the Southern Alps is Mt. Cook, at an elevation of 3754 meters. Its snow-covered peak is visible to the left of center in each of these MISR images. From the high peaks, glaciers have gouged long, slender mountain lakes and coastal fiords. Immediately to the southeast of Mt. Cook (to the right in these images), the glacial pale-blue water of Lake Pukaki stands out. Further to the south in adjacent valleys you can easily see Lakes Hawea and Wanaka, between which (though not visible here) is the Haast Pass Road, the most southerly of the few links between the east and west coast road systems. Further to the south is the prominent 'S' shape of Lake Wakatipu, 83 kilometers long, on the northern shore of which is Queenstown, the principal resort town of the island. The remote and spectacular Fiordland National

  1. Alkaline thermal sludge hydrolysis.

    PubMed

    Neyens, E; Baeyens, J; Creemers, C

    2003-02-28

    The waste activated sludge (WAS) treatment of wastewater produces excess sludge which needs further treatment prior to disposal or incineration. A reduction in the amount of excess sludge produced, and the increased dewaterability of the sludge are, therefore, subject of renewed attention and research. A lot of research covers the nature of the sludge solids and associated water. An improved dewaterability requires the disruption of the sludge cell structure. Previous investigations are reviewed in the paper. Thermal hydrolysis is recognized as having the best potential to meet the objectives and acid thermal hydrolysis is most frequently used, despite its serious drawbacks (corrosion, required post-neutralization, solubilization of heavy metals and phosphates, etc.). Alkaline thermal hydrolysis has been studied to a lesser extent, and is the subject of the detailed laboratory-scale research reported in this paper. After assessing the effect of monovalent/divalent cations (respectively, K(+)/Na(+) and Ca(2+)/Mg(2+)) on the sludge dewaterability, only the use of Ca(2+) appears to offer the best solution. The lesser effects of K(+), Na(+) and Mg(2+) confirm previous experimental findings. As a result of the experimental investigations, it can be concluded that alkaline thermal hydrolysis using Ca(OH)(2) is efficient in reducing the residual sludge amounts and in improving the dewaterability. The objectives are fully met at a temperature of 100 degrees C; at a pH approximately 10 and for a 60-min reaction time, where all pathogens are moreover killed. Under these optimum conditions, the rate of mechanical dewatering increases (the capillary suction time (CST) value is decreased from approximately 34s for the initial untreated sample to approximately 22s for the hydrolyzed sludge sample) and the amount of DS to be dewatered is reduced to approximately 60% of the initial untreated amount. The DS-content of the dewatered cake will be increased from 28 (untreated) to 46

  2. Nematode grazing promotes bacterial community dynamics in soil at the aggregate level

    PubMed Central

    Jiang, Yuji; Liu, Manqiang; Zhang, Jiabao; Chen, Yan; Chen, Xiaoyun; Chen, Lijun; Li, Huixin; Zhang, Xue-Xian; Sun, Bo

    2017-01-01

    Nematode predation has important roles in determining bacterial community composition and dynamics, but the extent of the effects remains largely rudimentary, particularly in natural environment settings. Here, we investigated the complex microbial–microfaunal interactions in the rhizosphere of maize grown in red soils, which were derived from four long-term fertilization regimes. Root-free rhizosphere soil samples were separated into three aggregate fractions whereby the abundance and community composition were examined for nematode and total bacterial communities. A functional group of alkaline phosphomonoesterase (ALP) producing bacteria was included to test the hypothesis that nematode grazing may significantly affect specific bacteria-mediated ecological functions, that is, organic phosphate cycling in soil. Results of correlation analysis, structural equation modeling and interaction networks combined with laboratory microcosm experiments consistently indicated that bacterivorous nematodes enhanced bacterial diversity, and the abundance of bacterivores was positively correlated with bacterial biomass, including ALP-producing bacterial abundance. Significantly, such effects were more pronounced in large macroaggregates than in microaggregates. There was a positive correlation between the most dominant bacterivores Protorhabditis and the ALP-producing keystone 'species' Mesorhizobium. Taken together, these findings implicate important roles of nematodes in stimulating bacterial dynamics in a spatially dependent manner. PMID:28742069

  3. Nematode grazing promotes bacterial community dynamics in soil at the aggregate level.

    PubMed

    Jiang, Yuji; Liu, Manqiang; Zhang, Jiabao; Chen, Yan; Chen, Xiaoyun; Chen, Lijun; Li, Huixin; Zhang, Xue-Xian; Sun, Bo

    2017-12-01

    Nematode predation has important roles in determining bacterial community composition and dynamics, but the extent of the effects remains largely rudimentary, particularly in natural environment settings. Here, we investigated the complex microbial-microfaunal interactions in the rhizosphere of maize grown in red soils, which were derived from four long-term fertilization regimes. Root-free rhizosphere soil samples were separated into three aggregate fractions whereby the abundance and community composition were examined for nematode and total bacterial communities. A functional group of alkaline phosphomonoesterase (ALP) producing bacteria was included to test the hypothesis that nematode grazing may significantly affect specific bacteria-mediated ecological functions, that is, organic phosphate cycling in soil. Results of correlation analysis, structural equation modeling and interaction networks combined with laboratory microcosm experiments consistently indicated that bacterivorous nematodes enhanced bacterial diversity, and the abundance of bacterivores was positively correlated with bacterial biomass, including ALP-producing bacterial abundance. Significantly, such effects were more pronounced in large macroaggregates than in microaggregates. There was a positive correlation between the most dominant bacterivores Protorhabditis and the ALP-producing keystone 'species' Mesorhizobium. Taken together, these findings implicate important roles of nematodes in stimulating bacterial dynamics in a spatially dependent manner.

  4. Systemic Manifestations in Pyridox(am)ine 5'-Phosphate Oxidase Deficiency.

    PubMed

    Guerriero, Réjean M; Patel, Archana A; Walsh, Brian; Baumer, Fiona M; Shah, Ankoor S; Peters, Jurriaan M; Rodan, Lance H; Agrawal, Pankaj B; Pearl, Phillip L; Takeoka, Masanori

    2017-11-01

    Pyridoxine is converted to its biologically active form pyridoxal-5-phosphate (P5P) by the enzyme pyridox(am)ine 5'-phosphate oxidase and serves as a cofactor in nearly 200 reactions in the central nervous system. Pyridox(am)ine 5'-phosphate oxidase deficiency leads to P5P dependent epilepsy, typically a neonatal- or infantile-onset epileptic encephalopathy treatable with P5P or in some cases, pyridoxine. Following identification of retinopathy in a patient with pyridox(am)ine 5'-phosphate oxidase deficiency that was reversible with P5P therapy, we describe the systemic manifestations of pyridox(am)ine 5'-phosphate oxidase deficiency. A series of six patients with homozygous mutations of PNPO, the gene coding pyridox(am)ine 5'-phosphate oxidase, were evaluated in our center over the course of two years for phenotyping of neurological and systemic manifestations. Five of six were born prematurely, three had anemia and failure to thrive, and two had elevated alkaline phosphatase. A movement disorder was observed in two children, and a reversible retinopathy was observed in the most severely affected infant. All patients had neonatal-onset epilepsy and were on a continuum of developmental delay to profound encephalopathy. Electroencephalographic features included background slowing and disorganization, absent sleep features, and multifocal and generalized epileptiform discharges. All the affected probands carried a homozygous PNPO mutation (c.674 G>T, c.686 G>A and c.352G>A). In addition to the well-described epileptic encephalopathy, pyridox(am)ine 5'-phosphate oxidase deficiency causes a range of neurological and systemic manifestations. A movement disorder, developmental delay, and encephalopathy, as well as retinopathy, anemia, and failure to thrive add to the broadening clinical spectrum of P5P dependent epilepsy. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Phosphate Solubilization Potentials of Rhizosphere Isolates from Central Anatolia (Turkey)

    NASA Astrophysics Data System (ADS)

    Ogut, M.; Er, F.

    2009-04-01

    Plant available-phosphorus (P) is usually low in Anatolian soils due mainly to the precipitation as calcium (Ca) and magnesium (Mg) phosphates in alkaline conditions. Phosphate solubilizing microorganisms (PSM) can enhance plant P-availability by dissolving the hardly soluble-P within the rhizosphere, which is the zone that surrounds the plant roots. PSM's can be used as seed- or soil-inocula to increase plant P-uptake and the overall growth. A total of 162 PSM's were isolated from the rhizosphere of wheat plants excavated from different fields located along a 75 km part of a highway in Turkey. The mean, the standart deviation, and the median for solubilized-P (ppm) in a 24 h culture in a tricalcium phosphate broth were 681, 427, and 400 for glucose; 358, 266, and 236 for sucrose; and 102, 117, and 50 for starch, respectively. There was not a linear relationship between the phosphate solubilized in the liquid cultures and the solubilization index obtained in the Pikovskaya's agar. Nine isolates representing both weak and strong solubilizers [Bacillus megaterium (5), Bacillus pumilis (1), Pseudomonas syringae pv. phaseolica (1), Pseudomonas fluorescens (1), Arthrobacter aurescens (1) as determined by the 16S rRNA gene sequence analysis] were further studied in a five day incubation. Pseudomonas syringae pv. phaseolica solubilized statistically (P<0.05) higher phosphate (409 ppm) than all the other strains did. There was not a statistically significant (P<0.05) difference in solubilized-P among the Bacillus strains. The pH of the medium fell to the levels between 4 and 5 from the initial neutrality. The phosphate solubilizing strains variably produced gluconic, 2-keto-D-gluconic, glycolic, acetic and butyric acids. The organic acids produced by these microorganisms seem to be the major source of phosphate solubilization in vitro.

  6. Phosphates in some Missouri refractory clays

    USGS Publications Warehouse

    Halley, Robert B.; Foord, Eugene E.; Keller, David J.; Keller, Walter D.

    1997-01-01

    host. The wavellite and variscite-strengite veinlets are secondary, precipitated from ion complexes in ground water percolating along cracks in the flint clay. The flint clay host of the variscite-strengite veinlet contains strontian crandallite. All of the phosphates contain significant amounts of strontium. The source of P, Ca and Sr was the marine carbonates. Dissolution of these carbonates produced the argillic residue that became the primordial Cheltenham paludal mud, which ultimately altered to fireclay.Preliminary firing tests show that the presence of phosphates lowers fusion temperature. However, it is not clear whether poor refractoriness is due to the presence of phosphates, per se, or to Ca, Sr and other alkaline elements present in the phosphates.

  7. Preparation of in situ hardening composite microcarriers: Calcium phosphate cement combined with alginate for bone regeneration

    PubMed Central

    Park, Jung-Hui; Lee, Eun-Jung; Knowles, Jonathan C

    2014-01-01

    Novel microcarriers consisting of calcium phosphate cement and alginate were prepared for use as three-dimensional scaffolds for the culture and expansion of cells that are effective for bone tissue engineering. The calcium phosphate cement-alginate composite microcarriers were produced by an emulsification of the composite aqueous solutions mixed at varying ratios (calcium phosphate cement powder/alginate solution = 0.8–1.2) in an oil bath and the subsequent in situ hardening of the compositions during spherodization. Moreover, a porous structure could be easily created in the solid microcarriers by soaking the produced microcarriers in water and a subsequent freeze-drying process. Bone mineral-like apatite nanocrystallites were shown to rapidly develop on the calcium phosphate cement–alginate microcarriers under moist conditions due to the conversion of the α-tricalcium phosphate phase in the calcium phosphate cement into a carbonate–hydroxyapatite. Osteoblastic cells cultured on the microspherical scaffolds were proven to be viable, with an active proliferative potential during 14 days of culture, and their osteogenic differentiation was confirmed by the determination of alkaline phosphatase activity. The in situ hardening calcium phosphate cement–alginate microcarriers developed herein may be used as potential three-dimensional scaffolds for cell delivery and tissue engineering of bone. PMID:23836845

  8. Influence of acidic and alkaline waste solution properties on uranium migration in subsurface sediments.

    PubMed

    Szecsody, Jim E; Truex, Mike J; Qafoku, Nikolla P; Wellman, Dawn M; Resch, Tom; Zhong, Lirong

    2013-08-01

    This study shows that acidic and alkaline wastes co-disposed with uranium into subsurface sediments have significant impact on changes in uranium retardation, concentration, and mass during downward migration. For uranium co-disposal with acidic wastes, significant rapid (i.e., hours) carbonate and slow (i.e., 100 s of hours) clay dissolution resulted, releasing significant sediment-associated uranium, but the extent of uranium release and mobility change was controlled by the acid mass added relative to the sediment proton adsorption capacity. Mineral dissolution in acidic solutions (pH2) resulted in a rapid (<10 h) increase in aqueous carbonate (with Ca(2+), Mg(2+)) and phosphate and a slow (100 s of hours) increase in silica, Al(3+), and K(+), likely from 2:1 clay dissolution. Infiltration of uranium with a strong acid resulted in significant shallow uranium mineral dissolution and deeper uranium precipitation (likely as phosphates and carbonates) with downward uranium migration of three times greater mass at a faster velocity relative to uranium infiltration in pH neutral groundwater. In contrast, mineral dissolution in an alkaline environment (pH13) resulted in a rapid (<10h) increase in carbonate, followed by a slow (10 s to 100 s of hours) increase in silica concentration, likely from montmorillonite, muscovite, and kaolinite dissolution. Infiltration of uranium with a strong base resulted in not only uranium-silicate precipitation (presumed Na-boltwoodite) but also desorption of natural uranium on the sediment due to the high ionic strength solution, or 60% greater mass with greater retardation compared with groundwater. Overall, these results show that acidic or alkaline co-contaminant disposal with uranium can result in complex depth- and time-dependent changes in uranium dissolution/precipitation reactions and uranium sorption, which alter the uranium migration mass, concentration, and velocity. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Asfotase-α improves bone growth, mineralization and strength in mouse models of neurofibromatosis type-1

    PubMed Central

    de la Croix Ndong, Jean; Makowski, Alexander James; Uppuganti, Sasidhar; Vignaux, Guillaume; Ono, Koichiro; Perrien, Daniel S.; Joubert, Simon; Baglio, Serena R.; Granchi, Donatella; Stevenson, David A.; Rios, Jonathan J.; Nyman, Jeffry S.; Elefteriou, Florent

    2014-01-01

    Mineralization of the skeleton depends on the balance between levels of pyrophosphate (PPi), an inhibitor of hydroxyapatite formation, and phosphate generated from PPi breakdown by alkaline phosphatase (ALP). We report here that ablation of Nf1, encoding the RAS/GTPase–activating protein neurofibromin, in bone–forming cells leads to supraphysiologic PPi accumulation, caused by a chronic ERK–dependent increase in genes promoting PPi synthesis and extracellular transport, namely Enpp1 and Ank. It also prevents BMP2–induced osteoprogenitor differentiation and, consequently, expression of ALP and PPi breakdown, further contributing to PPi accumulation. The short stature, impaired bone mineralization and strength in mice lacking Nf1 in osteochondroprogenitors or osteoblasts could be corrected by enzyme therapy aimed at reducing PPi concentration. These results establish neurofibromin as an essential regulator of bone mineralization, suggest that altered PPi homeostasis contributes to the skeletal dysplasiae associated with neurofibromatosis type-1 (NF1), and that some of the NF1 skeletal conditions might be preventable pharmacologically. PMID:24997609

  10. Near-infrared fluorescence probes for enzymes based on binding affinity modulation of squarylium dye scaffold.

    PubMed

    Oushiki, Daihi; Kojima, Hirotatsu; Takahashi, Yuki; Komatsu, Toru; Terai, Takuya; Hanaoka, Kenjiro; Nishikawa, Makiya; Takakura, Yoshinobu; Nagano, Tetsuo

    2012-05-15

    We present a novel design strategy for near-infrared (NIR) fluorescence probes utilizing dye-protein interaction as a trigger for fluorescence enhancement. The design principle involves modification of a polymethine dye with cleavable functional groups that reduce the dye's protein-binding affinity. When these functional groups are removed by specific interaction with the target enzymes, the dye's protein affinity is restored, protein binding occurs, and the dye's fluorescence is strongly enhanced. To validate this strategy, we first designed and synthesized an alkaline phosphatase (ALP) sensor by introducing phosphate into the squarylium dye scaffold; this sensor was able to detect ALP-labeled secondary antibodies in Western blotting analysis. Second, we synthesized a probe for β-galactosidase (widely used as a reporter of gene expression) by means of β-galactosyl substitution of the squarylium scaffold; this sensor was able to visualize β-galactosidase activity both in vitro and in vivo. Our strategy should be applicable to obtain NIR fluorescence probes for a wide range of target enzymes.

  11. The influence of different alkaline earth oxides on the structural and optical properties of undoped, Ce-doped, Sm-doped, and Sm/Ce co-doped lithium alumino-phosphate glasses

    NASA Astrophysics Data System (ADS)

    Othman, H. A.; Arzumanyan, G. M.; Möncke, D.

    2016-12-01

    Undoped, singly Sm doped, Ce doped, and Sm/Ce co-doped lithium alumino-phosphate glasses with different alkaline earth modifiers were prepared by melt quenching. The structure of the prepared glasses was investigated by FT-IR and Raman, as well as by optical spectroscopy. The effect of the optical basicity of the host glass matrix on the added active dopants was studied, as was the effect doping had on the phosphate structural units. The optical edge shifts toward higher wavelengths with an increase in the optical basicity due to the increased polarizability of the glass matrix, but also with increasing CeO2 concentration as a result of Ce3+/Ce4+ inter valence charge transfer (IV-CT) absorption. The optical band gap for direct and indirect allowed transitions was calculated for the undoped glasses. The glass sample containing Mg2+ modifier ions is found to have the highest value (4.16 eV) for the optical band gap while Ba2+ has the lowest value (3.61 eV). The change in the optical band gap arises from the structural changes and the overall polarizability (optical basicity). Refractive index, molar refractivity Rm and molar polarizability αm values increase with increasing optical basicity of the glasses. The characteristic absorption peaks of Sm3+ were also investigated. For Sm/Ce co-doped glasses, especially at high concentration of CeO2, the absorption of Ce3+ hinders the high energy absorption of Sm3+ and this effect becomes more obvious with increasing optical basicity.

  12. Interaction between Nitrogen and Phosphate Stress Responses in Sinorhizobium meliloti

    DOE PAGES

    Hagberg, Kelly L.; Yurgel, Svetlana N.; Mulder, Monika; ...

    2016-11-30

    Bacteria have developed various stress response pathways to improve their assimilation and allocation of limited nutrients, such as nitrogen and phosphate. While both the nitrogen stress response (NSR) and phosphate stress response (PSR) have been studied individually, there are few experiments reported that characterize effects of multiple stresses on one or more pathways in Sinorhizobium meliloti, a facultatively symbiotic, nitrogen-fixing bacteria. The PII proteins, GlnB and GlnK, regulate the NSR activity, but analysis of global transcription changes in a PII deficient mutant suggest that the S. meliloti PII proteins may also regulate the PSR. PII double deletion mutants grow verymore » slowly and pseudoreversion of the slow growth phenotype is common. In order to understand this phenomenon better, transposon mutants were isolated that had a faster growing phenotype. One mutation was in phoB, the response regulator for a two component regulatory system that is important in the PSR. phoB::Tn5 mutants had different phenotypes in the wild type compared to a PII deficient background. This led to the hypothesis that phosphate stress affects the NSR and conversely, that nitrogen stress affects the PSR. These results show that phosphate availability affects glutamine synthetase activity and expression, which are often used as indicators of NSR activity, but that nitrogen availability did not affect alkaline phosphatase activity and expression, which are indicators of PSR activity. Finally, we conclude that the NSR is co-regulated by nitrogen and phosphate, whereas the PSR does not appear to be co-regulated by nitrogen in addition to its known phosphate regulation.« less

  13. Interaction between Nitrogen and Phosphate Stress Responses in Sinorhizobium meliloti

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hagberg, Kelly L.; Yurgel, Svetlana N.; Mulder, Monika

    Bacteria have developed various stress response pathways to improve their assimilation and allocation of limited nutrients, such as nitrogen and phosphate. While both the nitrogen stress response (NSR) and phosphate stress response (PSR) have been studied individually, there are few experiments reported that characterize effects of multiple stresses on one or more pathways in Sinorhizobium meliloti, a facultatively symbiotic, nitrogen-fixing bacteria. The PII proteins, GlnB and GlnK, regulate the NSR activity, but analysis of global transcription changes in a PII deficient mutant suggest that the S. meliloti PII proteins may also regulate the PSR. PII double deletion mutants grow verymore » slowly and pseudoreversion of the slow growth phenotype is common. In order to understand this phenomenon better, transposon mutants were isolated that had a faster growing phenotype. One mutation was in phoB, the response regulator for a two component regulatory system that is important in the PSR. phoB::Tn5 mutants had different phenotypes in the wild type compared to a PII deficient background. This led to the hypothesis that phosphate stress affects the NSR and conversely, that nitrogen stress affects the PSR. These results show that phosphate availability affects glutamine synthetase activity and expression, which are often used as indicators of NSR activity, but that nitrogen availability did not affect alkaline phosphatase activity and expression, which are indicators of PSR activity. Finally, we conclude that the NSR is co-regulated by nitrogen and phosphate, whereas the PSR does not appear to be co-regulated by nitrogen in addition to its known phosphate regulation.« less

  14. The prevalence of and risk factors for acute mountain sickness in the Eastern and Western Alps.

    PubMed

    Mairer, Klemens; Wille, Maria; Burtscher, Martin

    2010-01-01

    Acute mountain sickness (AMS) is the most common condition of high altitude illnesses. Its prevalence varies between 15% and 80% depending on the speed of ascent, absolute altitude reached, and individual susceptibility. Additionally, we assumed that the more experienced mountaineers of the Western Alps are less susceptible to developing AMS than recreational mountaineers of the Eastern Alps or tourist populations. Therefore, the main goals of the present study were the collection of data regarding the AMS prevalence and triggers in both the Eastern and Western Alps using identical methods. A total of 162 mountaineers, 79 in the Eastern Alps (3454 m) and 83 in the Western Alps (3817 m) were studied on the morning after their first night at high altitude. A diagnosis of AMS was based on a Lake Louise Score (LLS) ≥4, the presence of headache, and at least one additional symptom. Thirty of 79 subjects (38.0%) suffered from AMS at 3454 m in the Eastern Alps as did 29 of 83 (34.9%) at 3817 m in the Western Alps. After adjustment for altitude, the prevalence in the Western Alps constituted 24.5%, which differed significantly (p = 0.04) from that found in the Eastern Alps. The lower mountaineering experience of mountaineers in the Eastern Alps turned out to be the only factor for explaining their higher AMS prevalence. Thus, expert advice by mountain guides or experienced colleagues could help to reduce the AMS risk in these subjects.

  15. Square-wave voltammetry assays for glycoproteins on nanoporous gold

    PubMed Central

    Pandey, Binod; Bhattarai, Jay K.; Pornsuriyasak, Papapida; Fujikawa, Kohki; Catania, Rosa; Demchenko, Alexei V.; Stine, Keith J.

    2014-01-01

    Electrochemical enzyme-linked lectinsorbent assays (ELLA) were developed using nanoporous gold (NPG) as a solid support for protein immobilization and as an electrode for the electrochemical determination of the product of the reaction between alkaline phosphatase (ALP) and p-aminophenyl phosphate (p-APP), which is p-aminophenol (p-AP). Glycoproteins or concanavalin A (Con A) and ALP conjugates were covalently immobilized onto lipoic acid self-assembled monolayers on NPG. The binding of Con A – ALP (or soybean agglutinin – ALP) conjugate to glycoproteins covalently immobilized on NPG and subsequent incubation with p-APP substrate was found to result in square-wave voltammograms whose peak difference current varied with the identity of the glycoprotein. NPG presenting covalently bound glycoproteins was used as the basis for a competitive electrochemical assay for glycoproteins in solution (transferrin and IgG). A kinetic ELLA based on steric hindrance of the enzyme-substrate reaction and hence reduced enzymatic reaction rate after glycoprotein binding is demonstrated using immobilized Con A–ALP conjugates. Using the immobilized Con A-ALP conjugate, the binding affinity of immunoglobulin G (IgG) was found to be 105 nM, and that for transferrin was found to be 650 nM. Minimal interference was observed in the presence of 5 mg mL−1 BSA as a model serum protein in both the kinetic and competitive ELLA. Inhibition studies were performed with methyl D-mannoside for the binding of TSF and IgG to Con A-ALP; IC50 values were found to be 90 μM and 286 μM, respectively. Surface coverages of proteins were estimated using solution depletion and the BCA protein concentration assay. PMID:24611035

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

    PubMed

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

    2012-02-01

    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 in patients undergoing coronary artery bypass grafting. A total of 63 patients undergoing coronary artery bypass grafting were enrolled and prospectively randomized. Bovine intestinal alkaline phosphatase (n=32) or placebo (n=31) was administered as an intravenous bolus followed by continuous infusion for 36 hours. The primary endpoint was to evaluate alkaline phosphatase levels in both groups and to find out if administration of bIAP to patients undergoing CABG would lead to endogenous alkaline phosphatase release. No significant adverse effects were identified in either group. In all the 32 patients of the bIAP-treated group, we found an initial rise of plasma alkaline phosphatase levels due to bolus administration (464.27±176.17 IU/L). A significant increase of plasma alkaline phosphatase at 4-6 hours postoperatively was observed (354.97±95.00 IU/L) as well. Using LHA inhibition, it was shown that this second peak was caused by the generation of tissue non specific alkaline phosphatase (TNSALP-type alkaline phosphatase). Intravenous bolus administration plus 8 hours continuous infusion of alkaline phosphatase in patients undergoing coronary artery bypass grafting with cardiopulmonary bypass results in endogenous alkaline phosphatase release. This endogenous alkaline phosphatase may play a role in the immune defense system.

  17. Design and applications of biomimetic anthraquinone dyes. Purification of calf intestinal alkaline phosphatase with immobilised terminal ring analogues of C.I. reactive blue 2.

    PubMed

    Lindner, N M; Jeffcoat, R; Lowe, C R

    1989-06-28

    A 330-fold one-step purification of alkaline phosphatase from a crude calf intestinal extract has been achieved using specific elution with inorganic phosphate (5 mM) from a purpose designed adsorbent comprising a terminal ring phosphonate analogue of C.I. Reactive Blue 2 coupled to Sepharose CL-6B-200. The resulting alkaline phosphatase preparation displayed a specific activity in excess of 1000 U/mg and was of equivalent purity to commercial "high purity" preparations as deduced by sodium dodecyl sulphate polyacrylamide gel electrophoresis and specific activity comparisons.

  18. Determination of alkaline phosphatase based on affinity adsorption solid-substrate room temperature phosphorimetry using rhodamine 6G-dibromoluciferin luminescent nanoparticle to label lectin and prediction of diseases.

    PubMed

    Liu, Jia-Ming; Liu, Zhen-Bo; Hu, Li-Xiang; He, Hang-Xia; Yang, Min-Lan; Zhou, Ping; Chen, Xin-Hua; Zheng, Min-Min; Zeng, Xiao-Yi; Xu, Yue-Long

    2006-10-15

    In the presence of heavy atom perturber LiAc, the silicon dioxide nanoparticle containing rhodamine 6G (R) and dibromoluciferin (D) (R-D-SiO(2)) can emit strong and stable solid-substrate room temperature phosphorescence signal of R (lambda(ex)/lambda(em)=481/648 nm) and D (lambda(ex)/lambda(em)=457/622 nm) on the surface of acetyl cellulose membrane (ACM). R-D-SiO(2) is used to label triticum vulgare lectin (WGA). Then two types of affinity adsorption reactions, R-D-SiO(2)-WGA- alkaline phosphatase (ALP) (direct method) and WGA-ALP-WGA-R-D-SiO(2) (sandwich method), are carried out on ACM. The conditions and the analytical characteristics for the determination of ALP using affinity adsorption solid-substrate room temperature phosphorimetry (AA-SS-RTP) were studied. For a 0.40-microl drop of sample, results show that the detection limits of the sandwich method are 0.16 ag spot(-1)(457/622 nm) and 0.17 ag spot(-1)(481/648 nm), and the detection limits of the direct method are 0.41 ag spot(-1) (457/622 nm) and 0.44 ag spot(-1) (481/648 nm). The contents of ALP in human serum correlated well with those obtained by enzyme-linked immunoassay. This study shows that AA-SS-RTP whether by the sandwich method or the direct method, can combine very well the characteristics of both high sensitivity of SS-RTP and specificity of the immunoreaction. Simultaneously, whether the phosphorescence excitation/emission wavelength of either R or D in R-D-SiO(2) is chosen to determine ALP, this can promote the agility and widen the adaptability of AA-SS-RTP.

  19. Salivary alkaline phosphatase and calcium in caries-active type II diabetes mellitus patients: An in vivo study

    PubMed Central

    Hegde, Mithra N.; Tahiliani, Divya; Shetty, Shilpa; Devadiga, Darshana

    2014-01-01

    Background: Diabetes Mellitus is a metabolic syndrome, affecting the oral health in various ways with dental caries being one of the most common problems encountered. Saliva is one of the most abundant secretions in the human body with a variety of natural protective and defence molecules bathing the oral cavity maintaining equilibrium. Its collection is easy and non-invasive. Aims: To compare and evaluate salivary alkaline phosphatase levels and calcium ion levels between caries active type II diabetes mellitus patients and non-diabetics. Materials and Methods: This study was carried out on caries-active age and gender matched 60 non-diabetic and 60 patients with known Type II diabetes mellitus subjects of age group 25-50 years with DMFT index >10. Saliva sample was collected to analyse for alkaline phosphatase enzyme and concentration of calcium ions using Agappe kits. Statistical Analysis: Student ‘t’ test was used to correlate the salivary electrolyte concentration in non- diabetic and diabetic patients with dental caries. A ‘P’ value of 0.05 or less was considered significant. Results are presented as mean ± standard deviation (X ± SD). Results: The alkaline phosphatase (ALP) activity in saliva was higher in diabetic patients when compared to that of non-diabetic patients with salivary calcium ions were significantly higher in non-diabetic individuals. Conclusion: Diabetes Mellitus patients are more prone to dental caries, hence require intervention to improve the quality of saliva. PMID:25395756

  20. The mechanism of hydrolysis of beta-glycerophosphate by kidney alkaline phosphatase.

    PubMed Central

    Ahlers, J

    1975-01-01

    1. To identify the functional groups that are involved in the conversion of beta-glycerophosphate by alkaline phosphatase (EC 3.1.3.1) from pig kidney, the kinetics of alkaline phosphatase were investigated in the pH range 6.6-10.3 at substrate concentrations of 3 muM-30 mM. From the plots of log VH+ against pH and log VH+/KH+m against pH one functional group with pK = 7.0 and two functional groups with pK = 9.1 were identified. These groups are involved in substrate binding. Another group with pK = 8.8 was found, which in its unprotonated form catalyses substrate conversion. 2. GSH inhibits the alkaline phosphatase reversibly and non-competitively by attacking the bound Zn(II). 3. The influence of the H+ concentration on the activation by Mg2+ ions of alkaline pig kidney phosphate was investigated between pH 8.4 and 10.0. The binding of substrate and activating Mg2+ ions occurs independently at all pH values between 8.4 and 10.0. The activation mechanism is not affected by the H+ concentration. The Mg2+ ions are bound by a functional group with a pK of 10.15. 4. A scheme is proposed for the reaction between enzyme, substrate, Mg2+ and H+ and the overall rate equation is derived. 5. The mechanism of substrate binding and splitting by the functional groups of the active centre is discussed on the basis of a model. Mg2+ seems to play a role as an autosteric effector. PMID:995

  1. Effect of altitude on brain intracellular pH and inorganic phosphate levels

    PubMed Central

    Shi, Xian-Feng; Carlson, Paul J.; Kim, Tae-Suk; Sung, Young-Hoon; Hellem, Tracy L.; Fiedler, Kristen K.; Kim, Seong-Eun; Glaeser, Breanna; Wang, Kristina; Zuo, Chun S.; Jeong, Eun-Kee; Renshaw, Perry F.; Kondo, Douglas G.

    2015-01-01

    Normal brain activity is associated with task-related pH changes. Although central nervous system syndromes associated with significant acidosis and alkalosis are well understood, the effects of less dramatic and chronic changes in brain pH are uncertain. One environmental factor known to alter brain pH is the extreme, acute change in altitude encountered by mountaineers. However, the effect of long-term exposure to moderate altitude has not been studied. The aim of this two-site study was to measure brain intracellular pH and phosphate-bearing metabolite levels at two altitudes in healthy volunteers, using phosphorus-31 magnetic resonance spectroscopy (31P-MRS). Increased brain pH and reduced inorganic phosphate (Pi) levels were found in healthy subjects who were long-term residents of Salt Lake City, UT (4720 ft/1438 m), compared with residents of Belmont, MA (20 ft/6 m). Brain intracellular pH at the altitude of 4720 ft was more alkaline than that observed near sea level. In addition, the ratio of inorganic phosphate to total phosphate signal also shifted toward lower values in the Salt Lake City region compared with the Belmont area. These results suggest that long-term residence at moderate altitude is associated with brain chemical changes. PMID:24768210

  2. 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).

    PubMed

    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 (p<0.01) increase in SCC, Fe, Zn, Co and LDH occurred in SCM milk containing gram positive bacterial agents only. ALP was found to be elevated in milk infected by both gram positive and negative bacteria. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and SCC≥2×10(5) cells/ml of milk as the benchmark. 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.

  3. Slab detachment under the Eastern Alps seen by seismic anisotropy

    PubMed Central

    Qorbani, Ehsan; Bianchi, Irene; Bokelmann, Götz

    2015-01-01

    We analyze seismic anisotropy for the Eastern Alpine region by inspecting shear-wave splitting from SKS and SKKS phases. The Eastern Alpine region is characterized by a breakdown of the clear mountain-chain-parallel fast orientation pattern that has been previously documented for the Western Alps and for the western part of the Eastern Alps. The main interest of this paper is a more detailed analysis of the anisotropic character of the Eastern Alps, and the transition to the Carpathian–Pannonian region. SK(K)S splitting measurements reveal a rather remarkable lateral change in the anisotropy pattern from the west to the east of the Eastern Alps with a transition area at about 12°E. We also model the backazimuthal variation of the measurements by a vertical change of anisotropy. We find that the eastern part of the study area is characterized by the presence of two layers of anisotropy, where the deeper layer has characteristics similar to those of the Central Alps, in particular SW–NE fast orientations of anisotropic axes. We attribute the deeper layer to a detached slab from the European plate. Comparison with tomographic studies of the area indicates that the detached slab might possibly connect with the lithosphere that is still in place to the west of our study area, and may also connect with the slab graveyard to the East, at the depth of the upper mantle transition zone. On the other hand, the upper layer has NW–SE fast orientations coinciding with a low-velocity layer which is found above a more-or-less eastward dipping high-velocity body. The anisotropy of the upper layer shows large-scale NW–SE fast orientation, which is consistent with the presence of asthenospheric flow above the detached slab foundering into the deeper mantle. PMID:25843968

  4. Slab detachment under the Eastern Alps seen by seismic anisotropy

    NASA Astrophysics Data System (ADS)

    Qorbani, Ehsan; Bianchi, Irene; Bokelmann, Götz

    2015-01-01

    We analyze seismic anisotropy for the Eastern Alpine region by inspecting shear-wave splitting from SKS and SKKS phases. The Eastern Alpine region is characterized by a breakdown of the clear mountain-chain-parallel fast orientation pattern that has been previously documented for the Western Alps and for the western part of the Eastern Alps. The main interest of this paper is a more detailed analysis of the anisotropic character of the Eastern Alps, and the transition to the Carpathian-Pannonian region. SK(K)S splitting measurements reveal a rather remarkable lateral change in the anisotropy pattern from the west to the east of the Eastern Alps with a transition area at about 12°E. We also model the backazimuthal variation of the measurements by a vertical change of anisotropy. We find that the eastern part of the study area is characterized by the presence of two layers of anisotropy, where the deeper layer has characteristics similar to those of the Central Alps, in particular SW-NE fast orientations of anisotropic axes. We attribute the deeper layer to a detached slab from the European plate. Comparison with tomographic studies of the area indicates that the detached slab might possibly connect with the lithosphere that is still in place to the west of our study area, and may also connect with the slab graveyard to the East, at the depth of the upper mantle transition zone. On the other hand, the upper layer has NW-SE fast orientations coinciding with a low-velocity layer which is found above a more-or-less eastward dipping high-velocity body. The anisotropy of the upper layer shows large-scale NW-SE fast orientation, which is consistent with the presence of asthenospheric flow above the detached slab foundering into the deeper mantle.

  5. Spectrophotometric determination of copper in alkaline solutions and evaluation of some hydroxy-substituted 1,10-phenanthrolines as chromogenic reagents.

    PubMed

    Dunbar, W E; Schilt, A A

    1972-09-01

    Seven new hydroxy-substituted 1,10-phenanthroline derivatives have been evaluated as chromogenic reagents for the determination of copper in strongly alkaline solution. The most sensitive of these, 2,9-dimethyl-4,7-dihydroxy-1,10-phenanthroline, has proven to be highly effective in a simple, rapid procedure for determining trace amounts of copper in sodium hydroxide, potassium carbonate, sodium phosphate or ammonium hydroxide.

  6. Regulation of the Synthesis of the Angucyclinone Antibiotic Alpomycin in Streptomyces ambofaciens by the Autoregulator Receptor AlpZ and Its Specific Ligand▿

    PubMed Central

    Bunet, Robert; Mendes, Marta V.; Rouhier, Nicolas; Pang, Xiuhua; Hotel, Laurence; Leblond, Pierre; Aigle, Bertrand

    2008-01-01

    Streptomyces ambofaciens produces an orange pigment and the antibiotic alpomycin, both of which are products of a type II polyketide synthase gene cluster identified in each of the terminal inverted repeats of the linear chromosome. Five regulatory genes encoding Streptomyces antibiotic regulatory proteins (alpV, previously shown to be an essential activator gene; alpT; and alpU) and TetR family receptors (alpZ and alpW) were detected in this cluster. Here, we demonstrate that AlpZ, which shows high similarity to γ-butyrolactone receptors, is at the top of a pathway-specific regulatory hierarchy that prevents synthesis of the alp polyketide products. Deletion of the two copies of alpZ resulted in the precocious production of both alpomycin and the orange pigment, suggesting a repressor role for AlpZ. Consistent with this, expression of the five alp-located regulatory genes and of two representative biosynthetic structural genes (alpA and alpR) was induced earlier in the alpZ deletion strain. Furthermore, recombinant AlpZ was shown to bind to specific DNA sequences within the promoter regions of alpZ, alpV, and alpXW, suggesting direct transcriptional control of these genes by AlpZ. Analysis of solvent extracts of S. ambofaciens cultures identified the existence of a factor which induces precocious production of alpomycin and pigment in the wild-type strain and which can disrupt the binding of AlpZ to its DNA targets. This activity is reminiscent of γ-butyrolactone-type molecules. However, the AlpZ-interacting molecule(s) was shown to be resistant to an alkali treatment capable of inactivating γ-butyrolactones, suggesting that the AlpZ ligand(s) does not possess a lactone functional group. PMID:18296523

  7. Regulation of the synthesis of the angucyclinone antibiotic alpomycin in Streptomyces ambofaciens by the autoregulator receptor AlpZ and its specific ligand.

    PubMed

    Bunet, Robert; Mendes, Marta V; Rouhier, Nicolas; Pang, Xiuhua; Hotel, Laurence; Leblond, Pierre; Aigle, Bertrand

    2008-05-01

    Streptomyces ambofaciens produces an orange pigment and the antibiotic alpomycin, both of which are products of a type II polyketide synthase gene cluster identified in each of the terminal inverted repeats of the linear chromosome. Five regulatory genes encoding Streptomyces antibiotic regulatory proteins (alpV, previously shown to be an essential activator gene; alpT; and alpU) and TetR family receptors (alpZ and alpW) were detected in this cluster. Here, we demonstrate that AlpZ, which shows high similarity to gamma-butyrolactone receptors, is at the top of a pathway-specific regulatory hierarchy that prevents synthesis of the alp polyketide products. Deletion of the two copies of alpZ resulted in the precocious production of both alpomycin and the orange pigment, suggesting a repressor role for AlpZ. Consistent with this, expression of the five alp-located regulatory genes and of two representative biosynthetic structural genes (alpA and alpR) was induced earlier in the alpZ deletion strain. Furthermore, recombinant AlpZ was shown to bind to specific DNA sequences within the promoter regions of alpZ, alpV, and alpXW, suggesting direct transcriptional control of these genes by AlpZ. Analysis of solvent extracts of S. ambofaciens cultures identified the existence of a factor which induces precocious production of alpomycin and pigment in the wild-type strain and which can disrupt the binding of AlpZ to its DNA targets. This activity is reminiscent of gamma-butyrolactone-type molecules. However, the AlpZ-interacting molecule(s) was shown to be resistant to an alkali treatment capable of inactivating gamma-butyrolactones, suggesting that the AlpZ ligand(s) does not possess a lactone functional group.

  8. The rising greenhouse effect: experiments and observations in and around the Alps

    NASA Astrophysics Data System (ADS)

    Philipona, R.

    2010-09-01

    The rapid temperature increase of more than 1°C in central Europe over the last three decades is larger than expected from anthropogenic greenhouse warming. Surface radiation flux measurements in and around the Alps in fact confirm that not only thermal longwave radiation but also solar shortwave radiation increased since the 1980s. Surface energy budget analyses reveal the rising surface temperature to be well correlated with the radiative forcing, and also show an increase of the kinetic energy fluxes explaining the rise of atmospheric water vapor. Solar radiation mainly increased due to a strong decline of anthropogenic aerosols since mid of the 1980s. While anthropogenic aerosols were mainly accumulated in the boundary layer, this reduction let solar radiation to recover (solar brightening after several decades of solar dimming) mainly at low altitudes around the Alps. At high elevations in the Alps, solar forcing is much smaller and the respective temperature rise is also found to be smaller than in the lowlands. The fact that temperature increases less in the Alps than at low elevations is unexpected in the concept of greenhouse warming, but the radiation budget analyses clearly shows that in the plains solar forcing due to declining aerosols additionally increased surface temperature, whereas in the Alps temperature increased primarily due to greenhouse warming that is particularly manifested by a strong water vapor feedback.

  9. ALP - blood test

    MedlinePlus

    ... Renal cell carcinoma Alternative Names Alkaline phosphatase References Berk PD, Korenblat KM. Approach to the patient with ... Review Date 5/21/2017 Updated by: Laura J. Martin, MD, MPH, ABIM Board Certified in Internal ...

  10. ALP isoenzyme test

    MedlinePlus

    ... Alkaline phosphatase isoenzyme test Images Blood test References Berk PD, Korenblat KM. Approach to the patient with ... Review Date 5/21/2017 Updated by: Laura J. Martin, MD, MPH, ABIM Board Certified in Internal ...

  11. No erosional control on the lateral growth of the Alps

    NASA Astrophysics Data System (ADS)

    Rosenberg, C. L.; Berger, A.

    2009-04-01

    On the base of literature data, we estimated the paleowidth of the Central Alps and the changing location of the inferred active fronts of the orogen from the Oligocene to the present. These compilations indicate that the absolute change of width, defined as the distance between the most external, but not necessarily active thrusts of the orogen was modest, amounting to less than 15 %, from 32 Ma to the present. This value lies within the error of estimate, and hence it is no sound evidence for lateral growth or retreat of the orogen. On the other hand the width of the active orogen, defined as the distance between the most external active thrusts, did increase in the early Miocene. This increase started already in the Oligocene as also concluded on the base of sedimentological findings, suggesting a continuous growth of thrusts through the Oligocene-Early Miocene interval (e.g. Schumacher et al., 1996). In the Late Miocene the active width of the Southern Alps decreased, as documented by a pre-Messinian out-of-sequence phase of thrusting (Lecco thrust; Schönborn, 1992) younger than the Milan Belt (Schönborn 1992). Increasing erosion rates are expected to reduce the width of the orogen, whereas decreasing rates are expected to increase its width (Beaumont et al., 1992). Therefore, following the example of previous investigations (Schlunegger et al., 2001; Schlunegger and Simpson, 2002; Willett et al., 2006) we compare the reconstructed changes of width of the Alps with the depositional budgets of the Alpine foreland basins (Kuhlemann, 2000) inferred to be a proxy for the erosion rates of the Alpine belt. This comparison shows that the most significant increases in erosion efficiency do not lead to a decrease in the active width of the orogen. This is indicated by the pronounced foreland-directed growth of the Alps after the Messinian, i.e., during the phase of greatest increase in the erosion rates of the orogen. The best regional examples are the northward shift

  12. Seismic anisotropy and large-scale deformation of the Eastern Alps

    NASA Astrophysics Data System (ADS)

    Bokelmann, Götz; Qorbani, Ehsan; Bianchi, Irene

    2013-12-01

    Mountain chains at the Earth's surface result from deformation processes within the Earth. Such deformation processes can be observed by seismic anisotropy, via the preferred alignment of elastically anisotropic minerals. The Alps show complex deformation at the Earth's surface. In contrast, we show here that observations of seismic anisotropy suggest a relatively simple pattern of internal deformation. Together with earlier observations from the Western Alps, the SKS shear-wave splitting observations presented here show one of the clearest examples yet of mountain chain-parallel fast orientations worldwide, with a simple pattern nearly parallel to the trend of the mountain chain. In the Eastern Alps, the fast orientations do not connect with neighboring mountain chains, neither the present-day Carpathians, nor the present-day Dinarides. In that region, the lithosphere is thin and the observed anisotropy thus resides within the asthenosphere. The deformation is consistent with the eastward extrusion toward the Pannonian basin that was previously suggested based on seismicity and surface geology.

  13. 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

    PubMed Central

    Jakka, Siva R. K.; Gong, Liang; Hasler, James; Banerjee, Rahul; Sheets, Joel J.; Narva, Kenneth; Blanco, Carlos A.

    2015-01-01

    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

  14. [Bone metabolism, biochemical markers of bone resorption and formation processes and interleukine 6 cytokin level during coeliac disease].

    PubMed

    Fekih, Monia; Sahli, Hela; Ben Mustapha, Nadia; Mestiri, Imen; Fekih, Moncef; Boubaker, Jalel; Kaabachi, Naziha; Sellami, Mohamed; Kallel, Lamia; Filali, Azza

    2013-01-01

    Celiac disease (CD) is characterized by a malabsorption syndrom. The bone anomalies are one of the principal complications of this disease. The osteoporosis frequency is high: 3.4% among patients having with CD versus 0.2% in the general population. To study the bone mineral density during the CD, to compare it to a control group and to determine the anomalies of biochemical markers of bone turn over and level of interleukin 6 cytokin (IL6) in these patients. All patients with CD have a measurement of bone mineral density by dual-energy x-ray absorptiometry (DXA), a biological exam with dosing calcemia, vitamin D, parathormone (PTH), the osteoblastic bone formation markers (serum osteocalcin, ALP phosphates alkaline), bone osteoclastic activity (C Télopeptide: CTX) and of the IL6. 42 patients were included, with a median age of 33.6 years. 52. 8% of the patients had a low level of D vitamine associated to a high level of PTH. An osteoporosis was noted in 21.5% of patients. No case of osteoporosis was detected in the control group. The mean level of the CTX, ostéocalcine and the IL6 was higher among patients having an osteoporosis or ostéopenia compared to patients with normal bone (p = 0,017). The factors associated with an bone loss (osteopenia or osteoporosis) were: an age > 30 years, a weight <50 kg, a level of ALP phosphates alkaline > 90 UI/ml, an hypo albuminemia < 40 g/l and a level of CTX higher than 1.2. Our study confirms the impact of the CD on the bone mineral statute. The relative risk to have an osteopenia or an osteoporosis was 5 in our series. The measurement of the osseous mineral density would be indicated among patients having a CD.

  15. Determinants of bone mineral density in patients on haemodialysis or peritoneal dialysis--a cross-sectional, longitudinal study.

    PubMed

    Nybo, Mads; Jespersen, Bente; Aarup, Michael; Ejersted, Charlotte; Hermann, Anne Pernille; Brixen, Kim

    2013-01-01

    The aim of the study was to identify biomarkers of alteration in bone mineral density (BMD) in patients on haemodialysis (HD) and peritoneal dialysis (PD). In a cross-sectional, longitudinal study dual-energy X-ray absorptiometry scans were performed in 146 HD-patients and 28 PD-patients. Follow-up after 14 months (mean) was conducted in 73 patients. As potential biomarkers we investigated parathyroid hormone (PTH), 25-hydroxy vitamin-D, ionised calcium, albumin, phosphate, and total alkaline phosphatases (t-ALP). Both groups of dialysis patients had lower BMD in the femoral neck (BMD(neck)) (P < 0.001) and forearm (BMD(forearm)) (P < 0.001) compared to healthy controls, but comparable BMD in the lumbar spine (BMD(spine)). BMD did not differ between dialysis types, but patients ever-treated with glucocorticoids had significantly lower BMD, while patients with polycystic kidney disease had higher BMD. BMD correlated with body weight, actual age, age at initiation of dialysis, duration of dialysis and levels of PTH and t-ALP. However, t-ALP only remained associated with low BMD(spine) after adjusting for other factors (P = 0.001). In the follow-up study all patients had decreased BMD in all three locations, but only for the lumbar spine there was a significant association between BMD and the bone markers t-ALP (P = 0.009) and PTH (P = 0.013). Both HD and PD patients have low BMD, and increased concentrations of t-ALP is associated BMD(spine) after adjustment, while PTH and t-ALP is associated with decrease in BMD(spine) over time. This substantiates the use of these biomarkers in both types of dialysis patients.

  16. The European Alps as an interrupter of the Earth's conductivity structures

    NASA Astrophysics Data System (ADS)

    Al-Halbouni, D.

    2013-07-01

    Joint interpretation of magnetotelluric and geomagnetic depth sounding results in the period range of 10-105 s in the Western European Alps offer new insights into the conductivity structure of the Earth's crust and mantle. This first large scale electromagnetic study in the Alps covers a cross-section from Germany to northern Italy and shows the importance of the alpine mountain chain as an interrupter of continuous conductors. Poor data quality due to the highly crystalline underground is overcome by Remote Reference and Robust Processing techniques and the combination of both electromagnetic methods. 3-D forward modeling reveals on the one hand interrupted dipping crustal conductors with maximum conductances of 4960 S and on the other hand a lithosphere thickening up to 208 km beneath the central Western Alps. Graphite networks arising from Palaeozoic sedimentary deposits are considered to be accountable for the occurrence of high conductivity and the distribution pattern of crustal conductors. The influence of huge sedimentary Molasse basins on the electromagnetic data is suggested to be minor compared with the influence of crustal conductors. Dipping direction (S-SE) and maximum angle (10.1°) of the northern crustal conductor reveal the main thrusting conditions beneath the Helvetic Alps whereas the existence of a crustal conductor in the Briançonnais supports theses about its belonging to the Iberian Peninsula. In conclusion the proposed model arisen from combined 3-D modeling of noise corrected electromagnetic data is able to explain the geophysical influence of various structural features in and around the Western European Alps and serves as a background for further upcoming studies.

  17. Magnetite solubility and phase stability in alkaline media at elevated temperatures

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ziemniak, S.E.; Jones, M.E.; Combs, K.E.S.

    Magnetite, Fe{sub 3}O{sub 4}, is the dominant oxide constituent of the indigenous corrosion layers that form on iron base alloys in high purity, high temperature water. The apparent simultaneous stability of two distinct oxidation states of iron in this metal oxide is responsible for its unique solubility behavior. The present work was undertaken to extend the experimental and theoretical bases for estimating solubilities of an iron corrosion product (Fe{sub 3}O{sub 4}/Fe(OH){sub 2}) over a broader temperature range and in the presence of complexing, pH-controlling reagents. These results indicate that a surface layer of ferrous hydroxide controls magnetite solubility behavior atmore » low temperatures in much the same manner as a surface layer of nickel(II) hydroxide was previously reported to control the low temperature solubility behavior of NiO. The importance of Fe(III) ion complexes implies not only that most previously-derived thermodynamic properties of the Fe(OH){sub 3}{sup {minus}} ion are incorrect, but that magnetite phase stability probably shifts to favor a sodium ferric hydroxyphosphate compound in alkaline sodium phosphate solutions at elevated temperatures. The test methodology involved pumping alkaline solutions of known composition through a bed of Fe{sub 3}O{sub 4} granules and analyzing the emerging solution for Fe. Two pH-controlling reagents were tested: sodium phosphate and ammonia. Equilibria for the following reactions were described in thermodynamic terms: (a) Fe(OH){sub 2}/Fe{sub 3}O{sub 4} dissolution and transformation, (b) Fe(II) and Fe(III) ion hydroxocomplex formation (hydrolysis), (c) Fe(II) ion amminocomplex formation, and (d) Fe(II) and Fe(III) ion phosphatocomplex formation. 36 refs.« less

  18. [Effect of carbon and nitrogen sources and complex B vitamins on the synthesis of alkaline protease by different strains of Bacillus mesentericus and Bacillus subtilis].

    PubMed

    Emtseva, T V

    1975-01-01

    The effect of different sources of carbon, nitrogen, amino acids and vitamins on the synthesis of alkaline proteases by the stock and mutant strains of Bacillus mesentericus and by the natural strain of Bacillus subtilis-12 has been investigated. The maximum synthesis of alkaline protease has been obtained in the media containing starch or its hydrolysates--dextrine and maltose as the carbon source. Ammonium phosphate and casein as the nitrogen source prove to be optimal for Bac. mesentericus and Bac. subtilis, respectively. Complex B vitamins added to the nutrient medium accelerate the enzyme synthesis 2.5-4-fold.

  19. Overexpression of {alpha}-catenin increases osteoblastic differentiation in mouse mesenchymal C3H10T1/2 cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kim, Dohee; Yang, Jae-Yeon; Shin, Chan Soo, E-mail: csshin@snu.ac.kr

    2009-05-15

    {alpha}- and {beta}-Catenin link cadherins to the actin-based cytoskeleton at adherens junctions and regulate cell-cell adhesion. Although roles of cadherins and canonical Wnt-/{beta}-catenin-signaling in osteoblastic differentiation have been extensively studied, the role of {alpha}-catenin is not known. Murine embryonic mesenchymal stem cells, C3H10T1/2 cells, were transduced with retrovirus encoding {alpha}-catenin (MSCV-{alpha}-catenin-HA-GFP). In the presence of Wnt-3A conditioned medium or osteogenic medium ({beta}-glycerol phosphate and ascorbic acid), cells overexpressing {alpha}-catenin showed enhanced osteoblastic differentiation as measured by alkaline phosphatase (ALP) staining and ALP activity assay compared to cells transduced with empty virus (MSCV-GFP). In addition, mRNA expression of osteocalcin and Runx2more » was significantly increased compared to control. Cell aggregation assay revealed that {alpha}-catenin overexpression has significantly increased cell-cell aggregation. However, cellular {beta}-catenin levels (total, cytoplasmic-nuclear ratio) and {beta}-catenin-TCF/LEF transcriptional activity did not change by overexpression of {alpha}-catenin. Knock-down of {alpha}-catenin using siRNA decreased osteoblastic differentiation as measured by ALP assay. These results suggest that {alpha}-catenin overexpression increases osteoblastic differentiation by increasing cell-cell adhesion rather than Wnt-/{beta}-catenin-signaling.« less

  20. The AlpArray Seismic Network: current status and next steps

    NASA Astrophysics Data System (ADS)

    Hetényi, György; Molinari, Irene; Clinton, John; Kissling, Edi

    2016-04-01

    The AlpArray initiative (http://www.alparray.ethz.ch) is a large-scale European collaboration to study the entire Alpine orogen at high resolution and in 3D with a large variety of geoscientific methods. The core element of the initiative is an extensive and dense broadband seismological network, the AlpArray Seismic Network (AASN), which complements the permanent seismological stations to ensure homogeneous coverage of the greater Alpine area. The some 260 temporary stations of the AlpArray Seismic Network are operated as a joint effort by a number of institutions from Austria, Bosnia-Herzegovina, Croatia, Czech Republic, France, Germany, Hungary, Italy, Slovakia and Switzerland. The first stations were installed in Spring 2015 and the full AASN is planned to be operational by early Summer 2016. In this poster we present the actual status of the deployment, the effort undertaken by the contributing groups, station performance, typical noise levels, best practices in installation as well as in data management, often encountered challenges, and planned next steps including the deployment of ocean bottom seismometers in the Ligurian Sea.

  1. Glacial isostatic uplift of the European Alps

    PubMed Central

    Mey, Jürgen; Scherler, Dirk; Wickert, Andrew D.; Egholm, David L.; Tesauro, Magdala; Schildgen, Taylor F.; Strecker, Manfred R.

    2016-01-01

    Following the last glacial maximum (LGM), the demise of continental ice sheets induced crustal rebound in tectonically stable regions of North America and Scandinavia that is still ongoing. Unlike the ice sheets, the Alpine ice cap developed in an orogen where the measured uplift is potentially attributed to tectonic shortening, lithospheric delamination and unloading due to deglaciation and erosion. Here we show that ∼90% of the geodetically measured rock uplift in the Alps can be explained by the Earth’s viscoelastic response to LGM deglaciation. We modelled rock uplift by reconstructing the Alpine ice cap, while accounting for postglacial erosion, sediment deposition and spatial variations in lithospheric rigidity. Clusters of excessive uplift in the Rhône Valley and in the Eastern Alps delineate regions potentially affected by mantle processes, crustal heterogeneity and active tectonics. Our study shows that even small LGM ice caps can dominate present-day rock uplift in tectonically active regions. PMID:27830704

  2. Glacial isostatic uplift of the European Alps.

    PubMed

    Mey, Jürgen; Scherler, Dirk; Wickert, Andrew D; Egholm, David L; Tesauro, Magdala; Schildgen, Taylor F; Strecker, Manfred R

    2016-11-10

    Following the last glacial maximum (LGM), the demise of continental ice sheets induced crustal rebound in tectonically stable regions of North America and Scandinavia that is still ongoing. Unlike the ice sheets, the Alpine ice cap developed in an orogen where the measured uplift is potentially attributed to tectonic shortening, lithospheric delamination and unloading due to deglaciation and erosion. Here we show that ∼90% of the geodetically measured rock uplift in the Alps can be explained by the Earth's viscoelastic response to LGM deglaciation. We modelled rock uplift by reconstructing the Alpine ice cap, while accounting for postglacial erosion, sediment deposition and spatial variations in lithospheric rigidity. Clusters of excessive uplift in the Rhône Valley and in the Eastern Alps delineate regions potentially affected by mantle processes, crustal heterogeneity and active tectonics. Our study shows that even small LGM ice caps can dominate present-day rock uplift in tectonically active regions.

  3. Bone Alkaline Phosphatase and Tartrate-Resistant Acid Phosphatase: Potential Co-regulators of Bone Mineralization.

    PubMed

    Halling Linder, Cecilia; Ek-Rylander, Barbro; Krumpel, Michael; Norgård, Maria; Narisawa, Sonoko; Millán, José Luis; Andersson, Göran; Magnusson, Per

    2017-07-01

    Phosphorylated osteopontin (OPN) inhibits hydroxyapatite crystal formation and growth, and bone alkaline phosphatase (BALP) promotes extracellular mineralization via the release of inorganic phosphate from the mineralization inhibitor inorganic pyrophosphate (PPi). Tartrate-resistant acid phosphatase (TRAP), produced by osteoclasts, osteoblasts, and osteocytes, exhibits potent phosphatase activity towards OPN; however, its potential capacity as a regulator of mineralization has not previously been addressed. We compared the efficiency of BALP and TRAP towards the endogenous substrates for BALP, i.e., PPi and pyridoxal 5'-phosphate (PLP), and their impact on mineralization in vitro via dephosphorylation of bovine milk OPN. TRAP showed higher phosphatase activity towards phosphorylated OPN and PPi compared to BALP, whereas the activity of TRAP and BALP towards PLP was comparable. Bovine milk OPN could be completely dephosphorylated by TRAP, liberating all its 28 phosphates, whereas BALP dephosphorylated at most 10 phosphates. OPN, dephosphorylated by either BALP or TRAP, showed a partially or completely attenuated phosphorylation-dependent inhibitory capacity, respectively, compared to native OPN on the formation of mineralized nodules. Thus, there are phosphorylations in OPN important for inhibition of mineralization that are removed by TRAP but not by BALP. In conclusion, our data indicate that both BALP and TRAP can alleviate the inhibitory effect of OPN on mineralization, suggesting a potential role for TRAP in skeletal mineralization. Further studies are warranted to explore the possible physiological relevance of TRAP in bone mineralization.

  4. Geological setting of the southern termination of Western Alps

    NASA Astrophysics Data System (ADS)

    d'Atri, Anna; Piana, Fabrizio; Barale, Luca; Bertok, Carlo; Martire, Luca

    2016-09-01

    A revision of the stratigraphic and tectonic setting of the southern termination of the Western Alps, at the junction of the Maritime Alps with the westernmost Ligurian Alps, is proposed. In response to the Alpine kinematic evolution, a number of tectonic units formed on the deformed palaeo-European continental margin and were arranged in a NW-SE striking anastomosed pattern along the north-eastern boundary of the Argentera Massif. Because these tectonic units often cut across the palaeogeographic subdivision of the Alpine literature and show only partial affinity with their distinctive stratigraphic features, new attributions are proposed. The Subbriançonnais domain is here intended as a "deformation zone", and its tectonic units have been attributed to Dauphinois and Provençal domains; furthermore, the Eocene Alpine Foreland Basin succession has been interpreted, based on the affinity of its lithologic characters and age, as a single feature resting above all the successions of the different Mesozoic domains. The Cretaceous tectono-sedimentary evolution of the studied domains was characterized by intense tectonic controls on sedimentation inducing lateral variations of stratigraphic features and major hydrothermal phenomena. Since the early Oligocene, transpressional tectonics induced a NE-SW shortening, together with significant left-lateral movements followed by (late Oligocene-middle Miocene) right-lateral movements along E-W to SE-NW striking shear zones. This induced the juxtaposition and/or stacking of Briançonnais, Dauphinois and Ligurian tectonic units characterized by different metamorphic histories, from anchizonal to lower greenschist facies. This evolution resulted in the arrangement of the tectonostratigraphic units in a wide "transfer zone" accommodating the Oligocene WNW-ward movement of portions of the palaeo-European margin placed at the south-western termination of Western Alps and the Miocene dextral shearing along SE striking faults that

  5. Kinetic behaviour of calf intestinal alkaline phosphatase with pNPP.

    PubMed

    Chaudhuri, Gouri; Chatterjee, Saswata; Venu-Babu, P; Ramasamy, K; Thilagaraj, W Richard

    2013-02-01

    The hydrolysis of p-nitrophenyl phosphate (pNPP) by calf intestinal alkaline phosphatase (CIAP) was investigated with respect to kinetic parameters such as V(max), K(m) and K(cat) under varying pH, buffers, substrate concentration, temperature and period of incubation. Highest activity was obtained with Tris-HCl at pH 11, while in the case of glycine-NaOH buffer the peak activity was recorded at pH 9.5. The enzyme showed the following kinetic characteristics with pNPP in 50 mM Tris-HCl at pH 11 and 100 mM glycine-NaOH at pH 9.5 at an incubation temperature of 37 degrees C: V(max), 3.12 and 1.6 micromoles min(-1) unit(-1); K(m), 7.6 x 10(-4) M and 4 x 10(-4) M; and K(cat), 82.98 s(-1) and 42.55 s(-1), respectively. CIAP displayed a high temperature optimum of 45 degrees C at pH 11. The kinetic behaviour of the enzyme under different parameters suggested that the enzyme might undergo subtle conformational changes in response to the buffers displaying unique characteristics. Bioprecipitation of Cu2+ from 50 ppm of CuCl2 solution was studied where 64.3% of precipitation was obtained. P(i) generated from CIAP-mediated hydrolysis of pNPP was found to bind with copper and precipitated as copper-phosphate. Thus, CIAP could be used as a test candidate in bioremediation of heavy metals from industrial wastes through generation of metal-phosphate complexes.

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

    PubMed Central

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

    2012-01-01

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

  7. Effect of micro-nano-hybrid structured hydroxyapatite bioceramics on osteogenic and cementogenic differentiation of human periodontal ligament stem cell via Wnt signaling pathway.

    PubMed

    Mao, Lixia; Liu, Jiaqiang; Zhao, Jinglei; Chang, Jiang; Xia, Lunguo; Jiang, Lingyong; Wang, Xiuhui; Lin, Kaili; Fang, Bing

    2015-01-01

    The surface structure of bioceramic scaffolds is crucial for its bioactivity and osteoinductive ability, and in recent years, human periodontal ligament stem cells have been certified to possess high osteogenic and cementogenic differential ability. In the present study, hydroxyapatite (HA) bioceramics with micro-nano-hybrid surface (mnHA [the hybrid of nanorods and microrods]) were fabricated via hydrothermal reaction of the α-tricalcium phosphate granules as precursors in aqueous solution, and the effects of mnHA on the attachment, proliferation, osteogenic and cementogenic differentiations of human periodontal ligament stem cells as well as the related mechanisms were systematically investigated. The results showed that mnHA bioceramics could promote cell adhesion, proliferation, alkaline phosphatase (ALP) activity, and expression of osteogenic/cementogenic-related markers including runt-related transcription factor 2 (Runx2), ALP, osteocalcin (OCN), cementum attachment protein (CAP), and cementum protein (CEMP) as compared to the HA bioceramics with flat and dense surface. Moreover, mnHA bioceramics stimulated gene expression of low-density lipoprotein receptor-related protein 5 (LRP5) and β-catenin, which are the key genes of canonical Wnt signaling. Moreover, the stimulatory effect on ALP activity and osteogenic and cementogenic gene expression, including that of ALP, OCN, CAP, CEMP, and Runx2 of mnHA bioceramics could be repressed by canonical Wnt signaling inhibitor dickkopf1 (Dkk1). The results suggested that the HA bioceramics with mnHA could act as promising grafts for periodontal tissue regeneration.

  8. Effect of micro-nano-hybrid structured hydroxyapatite bioceramics on osteogenic and cementogenic differentiation of human periodontal ligament stem cell via Wnt signaling pathway

    PubMed Central

    Mao, Lixia; Liu, Jiaqiang; Zhao, Jinglei; Chang, Jiang; Xia, Lunguo; Jiang, Lingyong; Wang, Xiuhui; Lin, Kaili; Fang, Bing

    2015-01-01

    The surface structure of bioceramic scaffolds is crucial for its bioactivity and osteoinductive ability, and in recent years, human periodontal ligament stem cells have been certified to possess high osteogenic and cementogenic differential ability. In the present study, hydroxyapatite (HA) bioceramics with micro-nano-hybrid surface (mnHA [the hybrid of nanorods and microrods]) were fabricated via hydrothermal reaction of the α-tricalcium phosphate granules as precursors in aqueous solution, and the effects of mnHA on the attachment, proliferation, osteogenic and cementogenic differentiations of human periodontal ligament stem cells as well as the related mechanisms were systematically investigated. The results showed that mnHA bioceramics could promote cell adhesion, proliferation, alkaline phosphatase (ALP) activity, and expression of osteogenic/cementogenic-related markers including runt-related transcription factor 2 (Runx2), ALP, osteocalcin (OCN), cementum attachment protein (CAP), and cementum protein (CEMP) as compared to the HA bioceramics with flat and dense surface. Moreover, mnHA bioceramics stimulated gene expression of low-density lipoprotein receptor-related protein 5 (LRP5) and β-catenin, which are the key genes of canonical Wnt signaling. Moreover, the stimulatory effect on ALP activity and osteogenic and cementogenic gene expression, including that of ALP, OCN, CAP, CEMP, and Runx2 of mnHA bioceramics could be repressed by canonical Wnt signaling inhibitor dickkopf1 (Dkk1). The results suggested that the HA bioceramics with mnHA could act as promising grafts for periodontal tissue regeneration. PMID:26648716

  9. Development of magnesium calcium phosphate biocement for bone regeneration.

    PubMed

    Jia, Junfeng; Zhou, Huanjun; Wei, Jie; Jiang, Xin; Hua, Hong; Chen, Fangping; Wei, Shicheng; Shin, Jung-Woog; Liu, Changsheng

    2010-08-06

    Magnesium calcium phosphate biocement (MCPB) with rapid-setting characteristics was fabricated by using the mixed powders of magnesium oxide (MgO) and calcium dihydrogen phosphate (Ca(H(2)PO(4))(2).H(2)O). The results revealed that the MCPB hardened after mixing the powders with water for about 7 min, and the compressive strength reached 43 MPa after setting for 1 h, indicating that the MCPB had a short setting time and high initial mechanical strength. After the acid-base reaction of MCPB containing MgO and Ca(H(2)PO(4))(2).H(2)O in a molar ratio of 2 : 1, the final hydrated products were Mg(3)(PO(4))(2) and Ca(3)(PO(4))(2). The MCPB was degradable in Tris-HCl solution and the degradation ratio was obviously higher than calcium phosphate biocement (CPB) because of its fast dissolution. The attachment and proliferation of the MG(63) cells on the MCPB were significantly enhanced in comparison with CPB, and the alkaline phosphatase activity of MG(63) cells on the MCPB was significantly higher than on the CPB at 7 and 14 days. The MG(63) cells with normal phenotype spread well on the MCPB surfaces, and were attached in close proximity to the substrate, as seen by scanning electron microscopy (SEM). The results demonstrated that the MCPB had a good ability to support cell attachment, proliferation and differentiation, and exhibited good cytocompatibility.

  10. Mineralogical, petrological and geochemical aspects of alkaline and alkaline-carbonatite associations from Brazil

    NASA Astrophysics Data System (ADS)

    Morbidelli, L.; Gomes, C. B.; Beccaluva, L.; Brotzu, P.; Conte, A. M.; Ruberti, E.; Traversa, G.

    1995-12-01

    A general description of Mesozoic and Tertiary (Fortaleza) Brazilian alkaline and alkaline-carbonatite districts is presented with reference to mineralogy, petrology, geochemistry and geochronology. It mainly refers to scientific results obtained during the last decade by an Italo-Brazilian research team. Alkaline occurrences are distributed across Brazilian territory from the southern (Piratini, Rio Grande do Sul State) to the northeastern (Fortaleza, Ceará State) regions and are mainly concentrated along the borders of the Paraná Basin generally coinciding with important tectonic lineaments. The most noteworthy characteristics of these alkaline and alkaline-carbonatite suites are: (i) prevalence of intrusive forms; (ii) abundance of cumulate assemblages (minor dunites, frequent clinopyroxenites and members of the ijolite series) and (iii) abundance of evolved rock-types. Many data demonstrate that crystal fractionation was the main process responsible for magma evolution of all Brazilian alkaline rocks. A hypothesis is proposed for the genesis of carbonatite liquids by immiscibility processes. The incidence of REE and trace elements for different major groups of lithotypes, belonging both to carbonatite-bearing and carbonatite-free districts, are documented. Sr and preliminary Nd isotopic data are indicative of a mantle origin for the least evolved magmas of all the studied occurrences. Mantle source material and melting models for the generation of the Brazilian alkaline magma types are also discussed.

  11. Alkaline "Permanent" Paper.

    ERIC Educational Resources Information Center

    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…

  12. Changes of forest cover and disturbance regimes in the mountain forests of the Alps.

    PubMed

    Bebi, P; Seidl, R; Motta, R; Fuhr, M; Firm, D; Krumm, F; Conedera, M; Ginzler, C; Wohlgemuth, T; Kulakowski, D

    2017-03-15

    Natural disturbances, such as avalanches, snow breakage, insect outbreaks, windthrow or fires shape mountain forests globally. However, in many regions over the past centuries human activities have strongly influenced forest dynamics, especially following natural disturbances, thus limiting our understanding of natural ecological processes, particularly in densely-settled regions. In this contribution we briefly review the current understanding of changes in forest cover, forest structure, and disturbance regimes in the mountain forests across the European Alps over the past millennia. We also quantify changes in forest cover across the entire Alps based on inventory data over the past century. Finally, using the Swiss Alps as an example, we analyze in-depth changes in forest cover and forest structure and their effect on patterns of fire and wind disturbances, based on digital historic maps from 1880, modern forest cover maps, inventory data on current forest structure, topographical data, and spatially explicit data on disturbances. This multifaceted approach presents a long-term and detailed picture of the dynamics of mountain forest ecosystems in the Alps. During pre-industrial times, natural disturbances were reduced by fire suppression and land-use, which included extraction of large amounts of biomass that decreased total forest cover. More recently, forest cover has increased again across the entire Alps (on average +4% per decade over the past 25-115 years). Live tree volume (+10% per decade) and dead tree volume (mean +59% per decade) have increased over the last 15-40 years in all regions for which data were available. In the Swiss Alps secondary forests that established after 1880 constitute approximately 43% of the forest cover. Compared to forests established previously, post-1880 forests are situated primarily on steep slopes (>30°), have lower biomass, a more aggregated forest structure (primarily stem-exclusion stage), and have been more strongly

  13. Synthesis and evaluation of thiophenyl derivatives as inhibitors of alkaline phosphatase.

    PubMed

    Chang, Lei; Duy, Do Le; Mébarek, Saida; Popowycz, Florence; Pellet-Rostaing, Stéphane; Lemaire, Marc; Buchet, René

    2011-04-15

    Pathological calcifications induced by deposition of basic phosphate crystals or hydroxyapatite (HA) on soft tissues are a large family of diseases comprising of ankylosing spondylitis (AS), end-stage osteoarthritis (OA) and vascular calcification. High activity of tissue non-specific alkaline phosphatase (TNAP) is a hallmark of pathological calcifications induced by HA deposition. The use of TNAP inhibitor is a possible therapeutic option to address calcific diseases produced by HA deposition on soft tissues. We report the synthesis of a series of thiopheno-imidazo[2,1-b]thiazole derivatives which were evaluated as potential inhibitors of TNAP displaying a large range of IC(50) at pH 10.4 (from 42±13 μM to more than 800 μM). Copyright © 2011. Published by Elsevier Ltd.

  14. Fingerprint deposition on nitrocellulose and polyvinylidene difluoride membranes using alkaline phosphatase.

    PubMed

    Kurien, Biji T; Danda, Debashish; Scofield, R Hal

    2015-01-01

    Dactyloscopy or fingerprint identification is a vital part of forensic evidence. Identification with fingerprints has been known since the finding of finger impressions on the clay surface of Babylonian legal contracts almost 4,000 years ago. The skin on the fingers and palms appears as grooves and ridges when observed under a microscope. A unique fingerprint is produced by the patterns of these friction skin ridges. Visible fingerprints can be deposited on solid surfaces. Colored inks have been used to deposit fingermarks on documents. Herein, we show that alkaline phosphatase can be used to transfer prints from fingers or palm to nitrocellulose or polyvinylidene difluoride membranes. The prints can be detected by using the nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate method of detection.

  15. Phosphate-Linked Silibinin Dimers (PLSd): New Promising Modified Metabolites.

    PubMed

    Romanucci, Valeria; Gravante, Raffaele; Cimafonte, Martina; Marino, Cinzia Di; Mailhot, Gilles; Brigante, Marcello; Zarrelli, Armando; Fabio, Giovanni Di

    2017-08-11

    By exploiting the regioselective protection of the hydroxyl groups of silibinin along with the well-known phosphoramidite chemistry, we have developed an efficient strategy for the synthesis of new silibinin-modified species, which we have named Phosphate-Linked Silibinin Dimers (PLSd), in which the monomer units are linked by phosphodiester bonds. The antioxidant abilities of the new PLSd were estimated on HepG2 cells using DPPH free radical scavenging and xanthine/xanthine oxidase assays. The new phosphate-metabolites showed a higher anti-oxidant activity than the silibinin, as well as very low toxicity. The ability to scavenge reactive oxygen species (ROS) such as singlet oxygen () and hydroxyl radical () reveals that the two dimers are able to scavenge about two times more effectively than silibinin. Finally, solubility studies have shown that the PLSd present good water solubility (more than 20 mg·L -1 ) under circumneutral pH values, whereas the silibinin was found to be very poorly soluble (less than 0.4 mg·L -1 ) and not stable under alkaline conditions. Together, the above promising results warrant further investigation of the future potential of the PLSd as anti-oxidant metabolites within the large synthetic polyphenols field.

  16. Characterization and Manipulation of the Pathway-Specific Late Regulator AlpW Reveals Streptomyces ambofaciens as a New Producer of Kinamycins ▿ †

    PubMed Central

    Bunet, Robert; Song, Lijiang; Mendes, Marta Vaz; Corre, Christophe; Hotel, Laurence; Rouhier, Nicolas; Framboisier, Xavier; Leblond, Pierre; Challis, Gregory L.; Aigle, Bertrand

    2011-01-01

    The genome sequence of Streptomyces ambofaciens, a species known to produce the congocidine and spiramycin antibiotics, has revealed the presence of numerous gene clusters predicted to be involved in the biosynthesis of secondary metabolites. Among them, the type II polyketide synthase-encoding alp cluster was shown to be responsible for the biosynthesis of a compound with antibacterial activity. Here, by means of a deregulation approach, we gained access to workable amounts of the antibiotics for structure elucidation. These compounds, previously designated as alpomycin, were shown to be known members of kinamycin family of antibiotics. Indeed, a mutant lacking AlpW, a member of the TetR regulator family, was shown to constitutively produce kinamycins. Comparative transcriptional analyses showed that expression of alpV, the essential regulator gene required for activation of the biosynthetic genes, is strongly maintained during the stationary growth phase in the alpW mutant, a stage at which alpV transcripts and thereby transcripts of the biosynthetic genes normally drop off. Recombinant AlpW displayed DNA binding activity toward specific motifs in the promoter region of its own gene and that of alpV and alpZ. These recognition sequences are also targets for AlpZ, the γ-butyrolactone-like receptor involved in the regulation of the alp cluster. However, unlike that of AlpZ, the AlpW DNA-binding ability seemed to be insensitive to the signaling molecules controlling antibiotic biosynthesis. Together, the results presented in this study reveal S. ambofaciens to be a new producer of kinamycins and AlpW to be a key late repressor of the cellular control of kinamycin biosynthesis. PMID:21193612

  17. Dried plum products as a substitute for phosphate in chicken marinade.

    PubMed

    Jarvis, Nathan; Clement, Ashley R; O'Bryan, Corliss A; Babu, Dinesh; Crandall, Philip G; Owens, Casey M; Meullenet, Jean-Francois; Ricke, Steven C

    2012-06-01

    In order to address the growing demand for more natural poultry products, alkaline phosphates commonly used in chicken breast meat marinades were replaced with plum ingredients and evaluated. For initial sensory evaluation, 200 consumers of chicken were served a small portion of the chicken breast on a plate and were asked to evaluate the product for overall impression, flavor, and texture on a 9-point hedonic scale with 1 = "dislike extremely" and 9 = "like extremely." Also, a 5-point just-about-right (JAR) scale was used on questions about tenderness, juiciness, overall flavor, and saltiness. Both hedonic and JAR demonstrated that the marinades of plum concentrate and the blend of plum fiber and powder were not distinguishable from the control (P > 0.05). Using two different percentages of fiber/powder blend, two different percentages of concentrate, sodium tripolyphosphate (STPP), and no marinade, measurements were made for marinade per cent pickup, thaw loss, and cook loss. Plum concentrate at 1.1% was most similar to STPP in marinade per cent pickup, thaw loss, drip loss, and cook loss. These results show that plum ingredients can potentially be used as a substitute in standard phosphate marinades. Consumers are increasingly demanding more natural foods with less artificial additives. This research presents the results of experiments using dried plum ingredients as a substitute for phosphates commonly used in marinades for chicken. Results indicate that dried plum ingredients may be a suitable substitute for phosphates in marinades. Journal of Food Science © 2012 Institute of Food Technologists® No claim to original US government works.

  18. P16/p53 expression and telomerase activity in immortalized human dental pulp cells

    PubMed Central

    Egbuniwe, Obi; Idowu, Bernadine D; Funes, Juan M; Grant, Andrew D; Renton, Tara

    2011-01-01

    Introduction Residing within human dental pulp are cells of an ectomesenchymal origin that have the potential to differentiate into odontoblast-like cells. These cells have a limited growth potential owing to the effects of cell senescence. This study examines the effects of immortalizing odontoblast-like cells on cell proliferation and mineralization by comparing transformed dental pulp stem cells (tDPSCs) and non-transformed dental pulp stem cells (nDPSCs). Results With the exogenous expression of hTERT, tDPSCs maintained a continued expression of odontogenic markers for cell proliferation and mineralization (ALP, COL-1, DMP-1, DSPP, OCN and OPN), as did nDPSCs. Oncoprotein expression was seen in both groups except for a noted absence of p16 in the tDPSCs. nDPSCs also showed lower levels of total ALP and DNA activity in comparison to tDPSCs when assayed, as well as low telomerase activity readings. Methods Using a retroviral vector, exogenous human telomerase reverse transcriptase (hTERT) was expressed in tDPSCs. Both cell groups were cultured, and their telomerase activities were determined using a telomerase quantification assay. Also examined, were the expression of genes involved in proliferation and mineralization, such as human alkaline phosphatase (ALP), β-actin, collagen I (col-1), core binding factor (cbfa)-1, dentin matrix protein (DMP-1), dentin sialophosphoprotein (DSPP), GAPDH, hTERT, osteocalcin (OCN), osteopontin (OPN) as well as oncoproteins involved in senescence (p16, p21 and p53) using RT-PCR. DNA and alkaline phosphate activity was also assayed in both cell groups. Conclusion These results indicate maintenance of odontoblast-like differentiation characteristics after retroviral transformation with hTERT and suggest a possible link with a reduced p16 expression. PMID:22067611

  19. Proteoliposomes harboring alkaline phosphatase and nucleotide pyrophosphatase as matrix vesicle biomimetics.

    PubMed

    Simão, Ana Maria S; Yadav, Manisha C; Narisawa, Sonoko; Bolean, Mayte; Pizauro, Joao Martins; Hoylaerts, Marc F; Ciancaglini, Pietro; Millán, José Luis

    2010-03-05

    We have established a proteoliposome system as an osteoblast-derived matrix vesicle (MV) biomimetic to facilitate the study of the interplay of tissue-nonspecific alkaline phosphatase (TNAP) and NPP1 (nucleotide pyrophosphatase/phosphodiesterase-1) during catalysis of biomineralization substrates. First, we studied the incorporation of TNAP into liposomes of various lipid compositions (i.e. in pure dipalmitoyl phosphatidylcholine (DPPC), DPPC/dipalmitoyl phosphatidylserine (9:1 and 8:2), and DPPC/dioctadecyl-dimethylammonium bromide (9:1 and 8:2) mixtures. TNAP reconstitution proved virtually complete in DPPC liposomes. Next, proteoliposomes containing either recombinant TNAP, recombinant NPP1, or both together were reconstituted in DPPC, and the hydrolysis of ATP, ADP, AMP, pyridoxal-5'-phosphate (PLP), p-nitrophenyl phosphate, p-nitrophenylthymidine 5'-monophosphate, and PP(i) by these proteoliposomes was studied at physiological pH. p-Nitrophenylthymidine 5'-monophosphate and PLP were exclusively hydrolyzed by NPP1-containing and TNAP-containing proteoliposomes, respectively. In contrast, ATP, ADP, AMP, PLP, p-nitrophenyl phosphate, and PP(i) were hydrolyzed by TNAP-, NPP1-, and TNAP plus NPP1-containing proteoliposomes. NPP1 plus TNAP additively hydrolyzed ATP, but TNAP appeared more active in AMP formation than NPP1. Hydrolysis of PP(i) by TNAP-, and TNAP plus NPP1-containing proteoliposomes occurred with catalytic efficiencies and mild cooperativity, effects comparable with those manifested by murine osteoblast-derived MVs. The reconstitution of TNAP and NPP1 into proteoliposome membranes generates a phospholipid microenvironment that allows the kinetic study of phosphosubstrate catabolism in a manner that recapitulates the native MV microenvironment.

  20. Proteoliposomes Harboring Alkaline Phosphatase and Nucleotide Pyrophosphatase as Matrix Vesicle Biomimetics*

    PubMed Central

    Simão, Ana Maria S.; Yadav, Manisha C.; Narisawa, Sonoko; Bolean, Mayte; Pizauro, Joao Martins; Hoylaerts, Marc F.; Ciancaglini, Pietro; Millán, José Luis

    2010-01-01

    We have established a proteoliposome system as an osteoblast-derived matrix vesicle (MV) biomimetic to facilitate the study of the interplay of tissue-nonspecific alkaline phosphatase (TNAP) and NPP1 (nucleotide pyrophosphatase/phosphodiesterase-1) during catalysis of biomineralization substrates. First, we studied the incorporation of TNAP into liposomes of various lipid compositions (i.e. in pure dipalmitoyl phosphatidylcholine (DPPC), DPPC/dipalmitoyl phosphatidylserine (9:1 and 8:2), and DPPC/dioctadecyl-dimethylammonium bromide (9:1 and 8:2) mixtures. TNAP reconstitution proved virtually complete in DPPC liposomes. Next, proteoliposomes containing either recombinant TNAP, recombinant NPP1, or both together were reconstituted in DPPC, and the hydrolysis of ATP, ADP, AMP, pyridoxal-5′-phosphate (PLP), p-nitrophenyl phosphate, p-nitrophenylthymidine 5′-monophosphate, and PPi by these proteoliposomes was studied at physiological pH. p-Nitrophenylthymidine 5′-monophosphate and PLP were exclusively hydrolyzed by NPP1-containing and TNAP-containing proteoliposomes, respectively. In contrast, ATP, ADP, AMP, PLP, p-nitrophenyl phosphate, and PPi were hydrolyzed by TNAP-, NPP1-, and TNAP plus NPP1-containing proteoliposomes. NPP1 plus TNAP additively hydrolyzed ATP, but TNAP appeared more active in AMP formation than NPP1. Hydrolysis of PPi by TNAP-, and TNAP plus NPP1-containing proteoliposomes occurred with catalytic efficiencies and mild cooperativity, effects comparable with those manifested by murine osteoblast-derived MVs. The reconstitution of TNAP and NPP1 into proteoliposome membranes generates a phospholipid microenvironment that allows the kinetic study of phosphosubstrate catabolism in a manner that recapitulates the native MV microenvironment. PMID:20048161

  1. The Effect of Acid Neutralization on Analytical Results Produced from SW846 Method 8330 after the Alkaline Hydrolysis of Explosives in Soil

    DTIC Science & Technology

    2012-09-01

    basic form of phosphoric acid or sodium phosphate NO2- Nitrite OH- Hydroxide ion ERDC/EL TR-12-14 1 1 Introduction Alkaline hydrolysis has...into amber sample vials and refrigerated until analyzed. TNT analyses were conducted by high performance liquid chromatography (HPLC) with a C-18...The explosives concentrations of the different soils were quantified using a DIONEX HPLC system equipped with a C-18 reverse phase column and a

  2. Enzyme replacement therapy for murine hypophosphatasia.

    PubMed

    Millán, José Luis; Narisawa, Sonoko; Lemire, Isabelle; Loisel, Thomas P; Boileau, Guy; Leonard, Pierre; Gramatikova, Svetlana; Terkeltaub, Robert; Camacho, Nancy Pleshko; McKee, Marc D; Crine, Philippe; Whyte, Michael P

    2008-06-01

    Hypophosphatasia (HPP) is the inborn error of metabolism that features rickets or osteomalacia caused by loss-of-function mutation(s) within the gene that encodes the tissue-nonspecific isozyme of alkaline phosphatase (TNALP). Consequently, natural substrates for this ectoenzyme accumulate extracellulary including inorganic pyrophosphate (PPi), an inhibitor of mineralization, and pyridoxal 5'-phosphate (PLP), a co-factor form of vitamin B6. Babies with the infantile form of HPP often die with severe rickets and sometimes hypercalcemia and vitamin B6-dependent seizures. There is no established medical treatment. Human TNALP was bioengineered with the C terminus extended by the Fc region of human IgG for one-step purification and a deca-aspartate sequence (D10) for targeting to mineralizing tissue (sALP-FcD10). TNALP-null mice (Akp2-/-), an excellent model for infantile HPP, were treated from birth using sALP-FcD10. Short-term and long-term efficacy studies consisted of once daily subcutaneous injections of 1, 2, or 8.2 mg/kg sALP-FcD10 for 15, 19, and 15 or 52 days, respectively. We assessed survival and growth rates, circulating levels of sALP-FcD10 activity, calcium, PPi, and pyridoxal, as well as skeletal and dental manifestations using radiography, microCT, and histomorphometry. Akp2-/- mice receiving high-dose sALP-FcD10 grew normally and appeared well without skeletal or dental disease or epilepsy. Plasma calcium, PPi, and pyridoxal concentrations remained in their normal ranges. We found no evidence of significant skeletal or dental disease. Enzyme replacement using a bone-targeted, recombinant form of human TNALP prevents infantile HPP in Akp2-/- mice.

  3. Revealing heterogeneous nucleation of primary Si and eutectic Si by AlP in hypereutectic Al-Si alloys.

    PubMed

    Li, Jiehua; Hage, Fredrik S; Liu, Xiangfa; Ramasse, Quentin; Schumacher, Peter

    2016-04-28

    The heterogeneous nucleation of primary Si and eutectic Si can be attributed to the presence of AlP. Although P, in the form of AlP particles, is usually observed in the centre of primary Si, there is still a lack of detailed investigations on the distribution of P within primary Si and eutectic Si in hypereutectic Al-Si alloys at the atomic scale. Here, we report an atomic-scale experimental investigation on the distribution of P in hypereutectic Al-Si alloys. P, in the form of AlP particles, was observed in the centre of primary Si. However, no significant amount of P was detected within primary Si, eutectic Si and the Al matrix. Instead, P was observed at the interface between the Al matrix and eutectic Si, strongly indicating that P, in the form of AlP particles (or AlP 'patch' dependent on the P concentration), may have nucleated on the surface of the Al matrix and thereby enhanced the heterogeneous nucleation of eutectic Si. The present investigation reveals some novel insights into heterogeneous nucleation of primary Si and eutectic Si by AlP in hypereutectic Al-Si alloys and can be used to further develop heterogeneous nucleation mechanisms based on adsorption.

  4. The onset of alpine pastoral systems in the Eastern Alps

    NASA Astrophysics Data System (ADS)

    Oeggl, Klaus; Festi, Daniela; Putzer, Andreas

    2015-04-01

    Since the discovery of the Neolithic glacier mummy "Ötzi" in the nival belt of the main Alpine ridge, the onset of alpine pasture is matter of a highly controversial debate both in archaeology and in palaeo-ecology of the Eastern Alps. The implication is that his sojourn in the high-altitudes of the Alps is considered to be connected with pastoral nomadism. Regrettably any archaeological evidence for the existence of such Neolithic alpine pastoral systems is missing up to now and the assumption is based on palynological data only. However, also the palynological record is ambiguous, because pasture indicators in the alpine regions react positive on grazing as well as on fertilization induced by a higher runoff of precipitation. Thus alpine pasture indicators reflect both grazing pressure and climatic change. Anyhow, alpine pastoral systems are a common practice in Alpine animal husbandry, but from an economic point of view such a seasonal vertical transhumance is costly. There are three main reasons for its practice: i) climatic, ii) economic (mainly in connection with population pressure or mining activities), and iii) cultural ideology. In this study we tested the above mentioned reasons in an interdisciplinary study on the beginning of pastoral activities in high altitudes in the central part of the Eastern Alps. This is conducted by palynological analyses of peat deposits situated in the vicinity of the timberline (1600 - 2400 m a.s.l.) combined with archaeological surveys. The investigated sites are located in traditional Alpine transhumance regions and aligned on a transect through the central part of the Eastern Alps. The studies reveal that grazing pressure is reflected since the Bronze Age, which is corroborated by archaeological findings in the vicinity of the investigated sites.

  5. Last Glacial Maximum Dated by Means of 10Be in the Maritime Alps (Italy)

    NASA Astrophysics Data System (ADS)

    Granger, D. E.; Spagnolo, M.; Federici, P.; Pappalardo, M.; Ribolini, A.; Cyr, A. J.

    2006-12-01

    Relatively few exposure dates of LGM moraines boulders are available for the European Alps, and none on the southern flank. Ponte Murato (PM) is a frontal moraine at 860 m asl in the Gesso Basin (Maritime Alps, SW European Alps). The PM moraine dams the 157 km2 Gesso della Barra Valley and it represents the lowermost frontal moraine of the entire Gesso Valley, near the outlet of the valley in the Po Plain. Its ELA, determined from the paleo- shape of the supposed Gesso della Barra glacier, is 1746 m asl. Tetti Bandito (TB) is a small and badly preserved glacial deposit, tentatively attributed to a lateral-frontal moraine, that is positioned 5 km downvalley from the PM deposit at 800 m asl. There are no other glacial deposits downvalley from the TB moraine in the Gesso Basin or farther NE in the piedmont region of the upper Po Plain. Boulders sampled on the PM and on the TB moraine crests gave a 10Be cosmogenic age of respectively 16300 ± 880 ka (average value) and 18798 ± 973 ka. This result constrains the PM frontal moraine within the LGM interval but also suggests that the maximum expansion of the Gesso Basin glacier was more downvalley at some point during the last glaciation. If the TB is a lateral-frontal moraine as supposed, the two TB and PM moraines would represent the outer and inner moraine crests of the same LGM stadial, with the outer moraine much less pronounced than the inner moraine, similarly to the maximalstand and the hochstand described in the Eastern Alps (Van Husen, 1997). Within this perspective, the PM and TB dates are consistent with a European Alps LGM corresponding to MIS 2 (Ivy-Ochs et al., 2004). This study of the Maritime Alps moraines is also in agreement with the Upper Würm climatic theory (Florineth and Schlüchter, 2000) of a stronger influence of the W and SW incoming humid airflows in the European Alps, differently from the nearby Vosges and Pyrenees mountain chains where more dry conditions were probably responsible for a very

  6. Dysregulation of phosphate metabolism and conditions associated with phosphate toxicity

    PubMed Central

    Brown, Ronald B; Razzaque, Mohammed S

    2015-01-01

    Phosphate homeostasis is coordinated and regulated by complex cross-organ talk through delicate hormonal networks. Parathyroid hormone (PTH), secreted in response to low serum calcium, has an important role in maintaining phosphate homeostasis by influencing renal synthesis of 1,25-dihydroxyvitamin D, thereby increasing intestinal phosphate absorption. Moreover, PTH can increase phosphate efflux from bone and contribute to renal phosphate homeostasis through phosphaturic effects. In addition, PTH can induce skeletal synthesis of another potent phosphaturic hormone, fibroblast growth factor 23 (FGF23), which is able to inhibit renal tubular phosphate reabsorption, thereby increasing urinary phosphate excretion. FGF23 can also fine-tune vitamin D homeostasis by suppressing renal expression of 1-alpha hydroxylase (1α(OH)ase). This review briefly discusses how FGF23, by forming a bone–kidney axis, regulates phosphate homeostasis, and how its dysregulation can lead to phosphate toxicity that induces widespread tissue injury. We also provide evidence to explain how phosphate toxicity related to dietary phosphorus overload may facilitate incidence of noncommunicable diseases including kidney disease, cardiovascular disease, cancers and skeletal disorders. PMID:26131357

  7. ALP conversion and the soft X-ray excess in the outskirts of the Coma cluster

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kraljic, David; Rummel, Markus; Conlon, Joseph P., E-mail: David.Kraljic@physics.ox.ac.uk, E-mail: Markus.Rummel@physics.ox.ac.uk, E-mail: j.conlon1@physics.ox.ac.uk

    2015-01-01

    It was recently found that the soft X-ray excess in the center of the Coma cluster can be fitted by conversion of axion-like-particles (ALPs) of a cosmic axion background (CAB) to photons. We extend this analysis to the outskirts of Coma, including regions up to 5 Mpc from the center of the cluster. We extract the excess soft X-ray flux from ROSAT All-Sky Survey data and compare it to the expected flux from ALP to photon conversion of a CAB. The soft X-ray excess both in the center and the outskirts of Coma can be simultaneously fitted by ALP tomore » photon conversion of a CAB. Given the uncertainties of the cluster magnetic field in the outskirts we constrain the parameter space of the CAB. In particular, an upper limit on the CAB mean energy and a range of allowed ALP-photon couplings are derived.« less

  8. Site-directed mutagenesis maps interactions that enhance cognate and limit promiscuous catalysis by an alkaline phosphatase superfamily phosphodiesterase.

    PubMed

    Wiersma-Koch, Helen; Sunden, Fanny; Herschlag, Daniel

    2013-12-23

    Catalytic promiscuity, an evolutionary concept, also provides a powerful tool for gaining mechanistic insights into enzymatic reactions. Members of the alkaline phosphatase (AP) superfamily are highly amenable to such investigation, with several members having been shown to exhibit promiscuous activity for the cognate reactions of other superfamily members. Previous work has shown that nucleotide pyrophosphatase/phosphodiesterase (NPP) exhibits a >10⁶-fold preference for the hydrolysis of phosphate diesters over phosphate monoesters, and that the reaction specificity is reduced 10³-fold when the size of the substituent on the transferred phosphoryl group of phosphate diester substrates is reduced to a methyl group. Here we show additional specificity contributions from the binding pocket for this substituent (herein termed the R' substituent) that account for an additional ~250-fold differential specificity with the minimal methyl substituent. Removal of four hydrophobic side chains suggested on the basis of structural inspection to interact favorably with R' substituents decreases phosphate diester reactivity 10⁴-fold with an optimal diester substrate (R' = 5'-deoxythymidine) and 50-fold with a minimal diester substrate (R' = CH₃). These mutations also enhance the enzyme's promiscuous phosphate monoesterase activity by nearly an order of magnitude, an effect that is traced by mutation to the reduction of unfavorable interactions with the two residues closest to the nonbridging phosphoryl oxygen atoms. The quadruple R' pocket mutant exhibits the same activity toward phosphate diester and phosphate monoester substrates that have identical leaving groups, with substantial rate enhancements of ~10¹¹-fold. This observation suggests that the Zn²⁺ bimetallo core of AP superfamily enzymes, which is equipotent in phosphate monoester and diester catalysis, has the potential to become specialized for the hydrolysis of each class of phosphate esters via addition

  9. Controlling the strontium-doping in calcium phosphate microcapsules through yeast-regulated biomimetic mineralization.

    PubMed

    Huang, Miaojun; Li, Tianjie; Pan, Ting; Zhao, Naru; Yao, Yongchang; Zhai, Zhichen; Zhou, Jiaan; Du, Chang; Wang, Yingjun

    2016-10-01

    Yeast cells have controllable biosorption on metallic ions during metabolism. However, few studies were dedicated to using yeast-regulated biomimetic mineralization process to control the strontium-doped positions in calcium phosphate microcapsules. In this study, the yeast cells were allowed to pre-adsorb strontium ions metabolically and then served as sacrificing template for the precipitation and calcination of mineral shell. The pre-adsorption enabled the microorganism to enrich of strontium ions into the inner part of the microcapsules, which ensured a slow-release profile of the trace element from the microcapsule. The co-culture with human marrow stromal cells showed that gene expressions of alkaline phosphatase and Collagen-I were promoted. The promotion of osteogenic differentiation was further confirmed in the 3D culture of cell-material complexes. The strategy using living microorganism as 'smart doping apparatus' to control incorporation of trace element into calcium phosphate paved a pathway to new functional materials for hard tissue regeneration.

  10. Alkaline volcanic rocks from the Columbia Hills, Gusev crater, Mars

    USGS Publications Warehouse

    McSween, H.Y.; Ruff, S.W.; Morris, R.V.; Bell, J.F.; Herkenhoff, K.; Gellert, Ralf; Stockstill, K.R.; Tornabene, L.L.; Squyres, S. W.; Crisp, J.A.; Christensen, P.R.; McCoy, T.J.; Mittlefehldt, D. W.; Schmidt, M.

    2006-01-01

    Irvine, Backstay, and Wishstone are the type specimens for three classes of fine-grained or fragmental, relatively unaltered rocks with distinctive thermal emission spectra, found as float on the flanks of the Columbia Hills. Chemical analyses indicate that these rocks are mildly alkaline basalt, trachybasalt, and tephrite, respectively. Their mineralogy consists of Na- and K-rich feldspar(s), low- and high-Ca pyroxenes, ferroan olivine, Fe-Ti (and possibly Cr) oxides, phosphate, and possibly glass. The texture of Wishstone is consistent with a pyroclastic origin, whereas Irvine and Backstay are lavas or possibly dike rocks. Chemical compositions of these rocks plot on or near liquid lines of descent for most elements calculated for Adirondack class rocks (olivine-rich basalts from the Gusev plains) at various pressures from 0.1 to 1.0 GPa. We infer that Wishstone-, Backstay-, and Irvine-class magmas may have formed by fractionation of primitive, oxidized basaltic magma similar to Adirondack-class rocks. The compositions of all these rocks reveal that the Gusev magmatic province is alkaline, distinct from the subalkaline volcanic rocks thought to dominate most of the planet's surface. The fact that differentiated volcanic rocks were not encountered on the plains prior to ascending Husband Hill may suggest a local magma source for volcanism beneath Gusev crater. Copyright 2006 by the American Geophysical Union.

  11. Reaching and abandoning the furthest ice extent during the Last Glacial Maximum in the Alps

    NASA Astrophysics Data System (ADS)

    Ivy-Ochs, Susan; Wirsig, Christian; Zasadni, Jerzy; Hippe, Kristina; Christl, Marcus; Akçar, Naki; Schluechter, Christian

    2016-04-01

    During the Last Glacial Maximum (LGM) in the European Alps (late Würm) local ice caps and extensive ice fields in the high Alps fed huge outlet glaciers that occupied the main valleys and extended onto the forelands as piedmont lobes. Records from numerous sites suggest advance of glaciers beyond the mountain front by around 30 ka (Ivy-Ochs 2015 and references therein). Reaching of the maximum extent occurred by about 27-26 ka, as exemplified by dates from the Rhein glacier area (Keller and Krayss, 2005). Abandonment of the outermost moraines at sites north and south of the Alps was underway by about 24 ka. In the high Alps, systems of transection glaciers with transfluences over many of the Alpine passes dominated, for example, at Grimsel Pass in the Central Alps (Switzerland). 10Be exposure ages of 23 ± 1 ka for glacially sculpted bedrock located just a few meters below the LGM trimline in the Haslital near Grimsel Pass suggest a pulse of ice surface lowering at about the same time that the foreland moraines were being abandoned (Wirsig et al., 2016). Widespread ice surface lowering in the high Alps was underway by no later than 18 ka. Thereafter, glaciers oscillated at stillstand and minor re-advance positions on the northern forelands for several thousand years forming the LGM stadial moraines. Final recession back within the mountain front took place by 19-18 ka. Recalculation to a common basis of all published 10Be exposure dates for boulders situated on LGM moraines suggests a strong degree of synchrony for the timing of onset of ice decay both north and south of the Alps. Ivy-Ochs, S., 2015, Cuadernos de investigación geográfica 41: 295-315. Keller, O., Krayss, E., 2005, Vierteljahrschr. Naturforsch. Gesell. Zürich 150: 69-85. Wirsig, C. et al., 2016, J. Quat. Sci. 31: 46-59.

  12. Quantifying the Eocene to Pleistocene topographic evolution of the southwestern Alps, France and Italy

    NASA Astrophysics Data System (ADS)

    Fauquette, Séverine; Bernet, Matthias; Suc, Jean-Pierre; Grosjean, Anne-Sabine; Guillot, Stéphane; van der Beek, Peter; Jourdan, Sébastien; Popescu, Speranta-Maria; Jiménez-Moreno, Gonzalo; Bertini, Adele; Pittet, Bernard; Tricart, Pierre; Dumont, Thierry; Schwartz, Stéphane; Zheng, Zhuo; Roche, Emile; Pavia, Giulio; Gardien, Véronique

    2015-02-01

    We evaluate the topographic evolution of the southwestern Alps using Eocene to Pleistocene pollen data combined with existing sedimentological, petrographic and detrital geo- and thermochronological data. We report 32 new pollen analyses from 10 sites completed by an existing dataset of 83 samples from 14 localities situated across the southwestern Alps, including both the pro- and the retro-foreland basins. The presence of microthermic tree pollen (mainly Abies, Picea) indicates that this part of the mountain belt attained elevations over 1900 m as early as the Oligocene. Inferred rapid surface uplift during the mid-Oligocene coincided with a previously documented brief phase of rapid erosional exhumation, when maximum erosion rates may have reached values of up to 1.5-2 km/Myr. Slower long-term average exhumation rates of ∼0.3 km/Myr since the Late Oligocene helped maintaining the high Alpine topography of the southwestern Alps until today. The relative abundances of meso-microthermic tree pollen (Cathaya, Cedrus and Tsuga) and microthermic tree pollen (Abies, Picea) in the pro- and retro-foreland basin deposits, indicate that the present-day asymmetric topography, with a relatively gentle western flank and steeper eastern flank, was established early in the southwestern Alps, at least since the Early Miocene, and possibly since the Oligocene or Late Eocene. Therefore, the high topography and asymmetric morphology of this part of the Alps has been maintained throughout the past ∼30 Ma.

  13. Nutrient and Trace Metal Controls on Alkaline Phosphatase in the Subtropical Ocean: Insights from Bioassays and Gene Expression

    NASA Astrophysics Data System (ADS)

    Mahaffey, C.; Reynolds, S.; Davis, C. E.; Lohan, M. C.

    2016-02-01

    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 in the surface ocean and limit biological activity. In response to phosphate limitation, organisms produce phosphohydrolytic enzymes, such as alkaline phosphatases (AP), that enable them to utilize the more replete dissolved organic phosphorus (DOP) pool to meet their cellular phosphorus demands. Synthesis of data from the surface ocean from 14 open ocean studies reveals an inverse hyperbolic relationship between phosphate and AP, where AP is significantly induced at phosphate concentrations below 50 nM and DOP concentrations decrease as AP increases. AP activity was 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 rates of AP, we demonstrate that other factors influence AP activity. AP are metalloenzymes and zinc and iron are co-factors of the AP proteins PhoA and PhoX, respectively. Using bioassay experiments, we show that the addition of Saharan dust and zinc significantly increases the rate of AP. To our knowledge, our results are the first direct field-based evidence that AP activity is limited by zinc in the subtropical ocean. In colonies of nitrogen fixer, Trichodesmium, we found enhanced expression of the phoA gene in a region of elevated zinc concentrations and enhanced expression of the phoX gene in a region of elevated iron concentrations around the intertropical convergence zone. Our study highlights the potential link between the phosphorus cycle and trace metals, specifically zinc and iron, and implies that there is potential for zinc-phosphorus and iron-phosphorus co-limitation in the ocean via AP.

  14. QUANTIFICATION OF GLACIAL EROSION IN THE ALPS USING VERY LOW-TEMPERATURE THERMOCHRONOLOGY (OSL & AHe)

    NASA Astrophysics Data System (ADS)

    Champagnac, J.; Herman, F.; Rhodes, E. J.; Fellin, M.; Jaiswal, M.; Schwenninger, J.; Reverman, R. L.

    2009-12-01

    The impact of glaciations on the topography of the Alps is still unclear: Long-term denudation rate determined by low-T thermochronology are in the range of 0.2 to 1 mm/yr, and increased during the Plio-Quaternary by 3 fold (Vernon et al., 2008). Such an increase is also documented by peri-alpine sediment budget (Kuhleman, 2000), with a similar increase in sediment yields since 5-3 Ma. This increase was considered as evidence of a climatically-driven surface process change, attributed to increased precipitation (Cederbom et al., 2004) and erosion by glacial processes (Champagnac et al., 2007). The timing of the onset of intense glacial erosion as well as its rates are still ambiguous. The glacial erosion seems to have accelerated around 0.9 Ma as suggested by the ten fold increase of incision rates of a valley in the Central Alps (Häuselmann et al., 2007), and by information about vegetation and sedimentologic changes (Muttoni et al., 2003). There is however no direct quantification of topographic change during the Plio-Quaternary. We present here how we use OSL-thermochronology, a new thermochronometer of exceptionally low closure temperature (about 30°-40°C) (Herman et al subm.), new {U-Th}/He on apatites data, and a glacial erosion model (Herman and Braun 2008) to estimate topographic changes in the Alps in response to glaciations. Because of their low closure temperature, OSL and AHe thermochronology enables quantification of events of less than 1 Ma at very small wavelength of the topography. We collected two vertical profiles, one in the Zermatt Valley (Valais) and one in Maurienne Valley (Savoy). We infer from these results changes in topography, date and quantify relief creation under glacial-interglacial cycles. Cederbom, C.E, et al., Climate induced rebound and exhumation of the European Alps. Geology 32, 709-712 (2000). Champagnac, J.-D., et al., Quaternary erosion-induced isostatic rebound in the western Alps. Geology 35, 195-198 (2007). Ha

  15. Study geomorphology, past and present, linear trench, tectonics relationship between Pyrenees and Alps

    NASA Technical Reports Server (NTRS)

    Guillemot, J. (Principal Investigator)

    1974-01-01

    The author has identified the following significant results. ERTS-1 images obviously show up some large linear features trending N 80 E or N 30 E common to both Alps and Pyrenees. One of them, the Ligurian Fault, had been previously forecast by Laubscher in an interpretation of the Alps by the plate tectonic theory, but it extends westward farthest from the Alps, cutting the Pyrenees axis. These lineaments have been interpreted as reflections of deep seated wrench faults in the surficial part of the sedimentary series. A large set of such lineaments is perceptible in western Europe, such as the Guadalquivir Fault in southern Spain, Ligurian Fault, Insubrian Fault, Northern-Jura Fault, Metz Fault. Perhaps these may be interpreted as transform faults of the mid-Atlantic ridge or of a paleo-rift seated in the Rhine-Rhone graben.

  16. Structurally controlled 'teleconnection' of large-scale mass wasting (Eastern Alps)

    NASA Astrophysics Data System (ADS)

    Ostermann, Marc; Sanders, Diethard

    2015-04-01

    In the Brenner Pass area (Eastern Alps) , closely ahead of the most northward outlier ('nose') of the Southern-Alpine continental indenter, abundant deep-seated gravitational slope deformations and a cluster of five post-glacial rockslides are present. The indenter of roughly triangular shape formed during Neogene collision of the Southern-Alpine basement with the Eastern-Alpine nappe stack. Compression by the indenter activated a N-S striking, roughly W-E extensional fault northward of the nose of the indenter (Brenner-normal fault; BNF), and lengthened the Eastern-Alpine edifice along a set of major strike-slip faults. These fault zones display high seismicity, and are the preferred locus of catastrophic rapid slope failures (rockslides, rock avalanches) and deep-seated gravitational slope deformations. The seismotectonic stress field, earthquake activity, and structural data all indicate that the South-Alpine indenter still - or again - exerts compression; in consequence, the northward adjacent Eastern Alps are subject mainly to extension and strike-slip. For the rockslides in the Brenner Pass area, and for the deep-seated gravitational slope deformations, the fault zones combined with high seismic activity predispose massive slope failures. Structural data and earthquakes mainly record ~W-E extension within an Eastern Alpine basement block (Oetztal-Stubai basement complex) in the hangingwall of the BNF. In the Northern Calcareous Alps NW of the Oetztal-Stubai basement complex, dextral faults provide defacement scars for large rockfalls and rockslides. Towards the West, these dextral faults merge into a NNW-SSE striking sinistral fault zone that, in turn, displays high seismic activity and is the locus of another rockslide cluster (Fern Pass cluster; Prager et al., 2008). By its kinematics dictated by the South-Alpine indenter, the relatively rigid Oetztal-Stubai basement block relays faulting and associated mass-wasting over a N-S distance of more than 60

  17. Orogen-scale anticline revealed in the Southern Alps of New Zealand by structural thermochronology

    NASA Astrophysics Data System (ADS)

    Zhou, Renjie; Brandon, Mark

    2017-04-01

    A dense set of cooling ages from the Southern Alps reveals an orogen-scale anticline of cooling-age isosurfaces (isochrones) and provides an interesting example of structural thermochronology, where isochrones are used as structural markers. The isochrone concept is an integral aspect of the age-elevation method, but the latter implicitly assumes that all isochrones are horizontal. Our experience in New Zeland and elsewhere is that isochrones are commonly tilted after formation. We use a more general approach that solves for orientation of the isochrone surfaces, and also the slope of the age-elevation trend, where "elevation" is measured normal to the isochrone surfaces. In New Zealand, collision and convergence between the Pacific and Australian plates have resulted in the formation and continuing growth of the Southern Alps, a prototypical orogenic wedge. In the western side, the Southern Alps is bounded by the Alpine fault, along with deeply exhumed rocks from depths up to 25 km. There are 150 apatite and 200 zircon fission-track (AFT, ZFT) ages that cover the vast region of the South Island of New Zealand from Lake Summer to Lake Wanaka. The AFT ages range from <0.5 to 140 Ma, and the ZFT ages, from <0.5 to 400 Ma. Our approach was initiated by McPhillips and Brandon (Earth and Planetary Science Letters, 2010, doi: 10.1016/j.epsl.2010.05.022). We use a least-squares method to solve for a best-fit sequence of dipping isochrone surfaces. The solution specifies the strike, dip and spacing of the parallel isochrones, the last of which indicates the velocity of the isochrones passing through the closure depth. We find that the calculation of the entire dataset failed to yield reasonable results, implying nonplanar structures at the regional scale. Using subsets of data, we observed three distinct zones of isochrones from E to W across the South Island. 1) The large area east of the Southern Alps in the central South Island contains ZFT isochrones that dip shallowly

  18. Antimicrobial properties and dentin bonding strength of magnesium phosphate cements.

    PubMed

    Mestres, G; Abdolhosseini, M; Bowles, W; Huang, S-H; Aparicio, C; Gorr, S-U; Ginebra, M-P

    2013-09-01

    The main objective of this work was to assess the antimicrobial properties and the dentin-bonding strength of novel magnesium phosphate cements (MPC). Three formulations of MPC, consisting of magnesium oxide and a phosphate salt, NH4H2PO4, NaH2PO4 or a mixture of both, were evaluated. As a result of the setting reaction, MPC transformed into either struvite (MgNH4PO4·6H2O) when NH4H2PO4 was used or an amorphous magnesium sodium phosphate when NaH2PO4 was used. The MPC had appropriate setting times for hard tissue applications, high early compressive strengths and higher strength of bonding to dentin than commercial mineral trioxide aggregate cement. Bacteriological studies were performed with fresh and aged cements against three bacterial strains, Escherichia coli, Pseudomonas aeruginosa (planktonic and in biofilm) and Aggregatibacter actinomycetemcomitans. These bacteria have been associated with infected implants, as well as other frequent hard tissue related infections. Extracts of different compositions of MPC had bactericidal or bacteriostatic properties against the three bacterial strains tested. This was associated mainly with a synergistic effect between the high osmolarity and alkaline pH of the MPC. These intrinsic antimicrobial properties make MPC preferential candidates for applications in dentistry, such as root fillers, pulp capping agents and cavity liners. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  19. 13 CFR 120.841 - Qualifications for the ALP.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    .... An applicant for ALP status must show that it substantially meets the following criteria: (a) CDC staff experience. The CDC's staff must have well-trained, qualified loan officers who are knowledgeable concerning SBA's lending policies and procedures for the 504 program. The CDC must have at least one loan...

  20. 13 CFR 120.841 - Qualifications for the ALP.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    .... An applicant for ALP status must show that it substantially meets the following criteria: (a) CDC staff experience. The CDC's staff must have well-trained, qualified loan officers who are knowledgeable concerning SBA's lending policies and procedures for the 504 program. The CDC must have at least one loan...

  1. 13 CFR 120.841 - Qualifications for the ALP.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    .... An applicant for ALP status must show that it substantially meets the following criteria: (a) CDC staff experience. The CDC's staff must have well-trained, qualified loan officers who are knowledgeable concerning SBA's lending policies and procedures for the 504 program. The CDC must have at least one loan...

  2. 13 CFR 120.841 - Qualifications for the ALP.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    .... An applicant for ALP status must show that it substantially meets the following criteria: (a) CDC staff experience. The CDC's staff must have well-trained, qualified loan officers who are knowledgeable concerning SBA's lending policies and procedures for the 504 program. The CDC must have at least one loan...

  3. Production of alkaline protease from Cellulosimicrobium cellulans

    PubMed Central

    Ferracini-Santos, Luciana; Sato, Hélia H

    2009-01-01

    Cellulosimicrobium cellulans is one of the microorganisms that produces a wide variety of yeast cell wall-degrading enzymes, β-1,3-glucanase, protease and chitinase. Dried cells of Saccharomyces cerevisiae were used as carbon and nitrogen source for cell growth and protease production. The medium components KH2PO4, KOH and dried yeast cells showed a significant effect (p<0.05) on the factorial fractional design. A second design was prepared using two factors: pH and percentage of dried yeast cells. The results showed that the culture medium for the maximum production of protease was 0.2 g/l of MgSO4.7H2O, 2.0 g/l of (NH4)2SO4 and 8% of dried yeast cells in 0.15M phosphate buffer at pH 8.0. The maximum alkaline protease production was 7.0 ± 0.27 U/ml over the center point. Crude protease showed best activity at 50ºC and pH 7.0-8.0, and was stable at 50ºC. PMID:24031317

  4. Cutting-edge Kinetic Physics with Parker Solar Probe and Solar Orbiter: The Arbitrary Linear Plasma Solver (ALPS)

    NASA Astrophysics Data System (ADS)

    Verscharen, D.; Klein, K. G.; Chandran, B. D. G.; Stevens, M. L.; Salem, C. S.; Bale, S. D.

    2017-12-01

    The Arbitrary Linear Plasma Solver (ALPS) is a parallelized numerical code that solves the dispersion relation in a hot (even relativistic) magnetized plasma with an arbitrary number of particle species with arbitrary gyrotropic equilibrium distribution functions for any direction of wave propagation with respect to the background field. In this way, ALPS retains generality and overcomes the shortcomings of previous (bi-)Maxwellian solvers for the plasma dispersion relations. The unprecedented high-resolution particle and field data products from Parker Solar Probe (PSP) and Solar Orbiter (SO) will require novel theoretical tools. ALPS is one such tool, and its use will make possible new investigations into the role of non-Maxwellian distributions in the near-Sun solar wind. It can be applied to numerous high-velocity-resolution systems, ranging from current space missions to numerical simulations. We will briefly discuss the ALPS algorithm and demonstrate its functionality based on previous solar-wind measurements. We will then highlight our plans for future applications of ALPS to PSP and SO observations.

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

    PubMed Central

    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

  6. Improved osteoblasts growth on osteomimetic hydroxyapatite/BaTiO3 composites with aligned lamellar porous structure.

    PubMed

    Liu, Beilei; Chen, Liangjian; Shao, Chunsheng; Zhang, Fuqiang; Zhou, Kechao; Cao, Jun; Zhang, Dou

    2016-04-01

    Osteoblasts growing into bone substitute is an important step of bone regeneration. This study prepared porous hydroxyapatite (HA)/BaTiO3 piezoelectric composites with porosity of 40%, 50% and 60% by ice-templating method. Effects of HA/BaTiO3 composites with different porosities, with and without polarizing treatment on adhesion, proliferation and differentiation of osteoblasts were investigated in vitro. Results revealed that cell densities of the porous groups were significantly higher than those of the dense group (p<0.05), so did the alkaline phosphate (ALP) and bone gla protein (BGP) activities. Porosity of 50% group exhibited higher ALP activity and BGP activity than those of the 40% and 60% groups. Scanning electron microscopy (SEM) observations revealed that osteoblasts adhered and stretched better on porous HA/BaTiO3 than on the dense one, especially HA/BaTiO3 with porosity of 50% and 60%. However, there was no significant difference in the cell morphology, cell densities, ALP and BGP activities between the polarized group and the non-polarized group (p>0.05). The absence of mechanical loading on the polarized samples may account for this. The results indicated that hierarchically porous HA/BaTiO3 played a favorable part in osteoblasts proliferation, differentiation and adhesion process and is a promising bone substitute material. Copyright © 2015. Published by Elsevier B.V.

  7. Use of Rituximab for Refractory Cytopenias Associated with Autoimmune Lymphoproliferative Syndrome (ALPS)

    PubMed Central

    Rao, V. Koneti; Price, Susan; Perkins, Katie; Aldridge, Patricia; Tretler, Jean; Davis, Joie; Dale, Janet K.; Gill, Fred; Hartman, Kip R.; Stork, Linda C.; Gnarra, David J.; Krishnamurti, Lakshmanan; Newburger, Peter E.; Puck, Jennifer; Fleisher, Thomas

    2009-01-01

    Background ALPS is a disorder of apoptosis resulting in accumulation of autoreactive lymphocytes, leading to marked lymphadenopathy, hepatosplenomegaly and multilineage cytopenias due to splenic sequestration and/or autoimmune destruction often presenting in childhood. We summarize our experience of rituximab use during the last 8 years in twelve patients, 9 children and 3 adults, out of 259 individuals with ALPS, belonging to 166 families currently enrolled in studies at the National Institutes of Health. Methods Refractory immune thrombocytopenia (platelet count <20,000) in 9 patients and autoimmune hemolytic anemia (AIHA) in 3 patients led to treatment with rituximab. Among them, 7 patients had undergone prior surgical splenectomy; 3 had significant splenomegaly; and 2 had no palpable spleen. Results In 7 out of 9 patients with ALPS and thrombocytopenia, rituximab therapy led to median response duration of 21months (range 14–36 months). In contrast, none of the 3 children treated with rituximab for AIHA responded. Noted toxicities included profound and prolonged hypogammaglobulinemia in 3 patients requiring replacement IVIG, total absence of antibody response to polysaccharide vaccines lasting up to 4 years after rituximab infusions in 1 patient and prolonged neutropenia in 1 patient. Conclusion Toxicities including hypogammaglobulinemia and neutropenia constitute an additional infection risk burden, especially in asplenic individuals, and may warrant avoidance of rituximab until other immunosuppressive medication options are exhausted. Long term follow up of ALPS patients with cytopenias after any treatment is necessary to determine relative risks and benefits. PMID:19214977

  8. Structure-solubility relationships in fluoride-containing phosphate based bioactive glasses

    NASA Astrophysics Data System (ADS)

    Shaharyar, Yaqoot

    The dissolution of fluoride-containing bioactive glasses critically affects their biomedical applications. Most commercial fluoride-releasing bioactive glasses have been designed in the soda-lime-silica system. However, their relatively slow chemical dissolution and the adverse effect of fluoride on their bioactivity are stimulating the study of novel biodegradable materials with higher bioactivity, such as biodegradable phosphate-based bioactive glasses, which can be a viable alternative for applications where a fast release of active ions is sought. In order to design new biomaterials with controlled degradability and high bioactivity, it is essential to understand the connection between chemical composition, molecular structure, and solubility in physiological fluids.Accordingly, in this work we have combined the strengths of various experimental techniques with Molecular Dynamics (MD) simulations, to elucidate the impact of fluoride ions on the structure and chemical dissolution of bioactive phosphate glasses in the system: 10Na2O - (45-x) CaO - 45P2O5 - xCaF2, where x varies between 0 -- 10 mol.%. NMR and MD data reveal that the medium-range atomic-scale structure of thse glasses is dominated by Q2 phosphate units followed by Q1 units, and the MD simulations further show that fluoride tends to associate with network modifier cations to form alkali/alkaline-earth rich ionic aggregates. On a macroscopic scale, we find that incorporating fluoride in phosphate glasses does not affect the rate of apatite formation on the glass surface in simulated body fluid (SBF). However, fluoride has a marked favorable impact on the glass dissolution in deionized water. Similarly, fluoride incorporation in the glasses results in significant weight gain due to adsorption of water (in the form of OH ions). These macroscopic trends are discussed on the basis of the F effect on the atomistic structure of the glasses, such as the F-induced phosphate network re-polymerization, in a

  9. Effects of atmospheric and climate change at the timberline of the Central European Alps

    PubMed Central

    Wieser, Gerhard; Matyssek, Rainer; Luzian, Roland; Zwerger, Peter; Pindur, Peter; Oberhuber, Walter; Gruber, Andreas

    2011-01-01

    This review considers potential effects of atmospheric change and climate warming within the timberline ecotone of the Central European Alps. After focusing on the impacts of ozone (O3) and rising atmospheric CO2 concentration, effects of climate warming on the carbon and water balance of timberline trees and forests will be outlined towards conclusions about changes in tree growth and treeline dynamics. Presently, ambient ground-level O3 concentrations do not exert crucial stress on adult conifers at the timberline of the Central European Alps. In response to elevated atmospheric CO2 Larix decidua showed growth increase, whereas no such response was found in Pinus uncinata. Overall climate warming appears as the factor responsible for the observed growth stimulation of timberline trees. Increased seedling re-establishment in the Central European Alps however, resulted from invasion into potential habitats rather than upward migration due to climate change, although seedlings will only reach tree size upon successful coupling with the atmosphere and thus loosing the beneficial microclimate of low stature vegetation. In conclusion, future climate extremes are more likely than the gradual temperature increase to control treeline dynamics in the Central European Alps. PMID:21379395

  10. Chemiluminescence-based pesticide biosensor utilizing the intelligent evolved properties of the enzyme alkaline phosphatase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ayyagari, M.; Kamtekar, S.; Pande, R.

    A methodology is described for immobilizing the enzyme alkaline phosphatase onto a glass surface using a novel biotinylated copolymer, poly(3-undecylthiophene-co-3- methanoithiophene). A streptavidin conjugate of alkaline phosphatase is used in this study. The biotinylated polymer is attached to the silanized glass surface via hydrophobic interactions and the enzyme is interfaced with the polymer through the classical biotin- streptavidin interaction. Alkaline phosphatase catalyzes the dephosphorylation of a macrocyclic compound, chloro-3-(4-methoxy spiro) (1,2 dioxetane-3-2`-tricyclo-) (3.3.1.1 )-(decani-4-yl) phenyl phosphate, to a species which emits energy by chemiluminescence. This chemiluminescence signal can be detected with a photomultiplier tube for enzymatic catalysis with the biocatalystmore » both in solution and immobilized on a glass surface. The signal generation is inhibited by the organophosphorus based insecticides such as paraoxon as well as nerve agents. We demonstrate in this study that a number of organophosphorus based insecticides inhibit the enzyme-mediated generation of chemiluminescence signal. This is true for the enzyme conjugate both free in solution and immobilized on a glass surface. In solution, the inhibition resembles the case of a partially uncompetitive system. By this type of inhibition we are able to detect pesticides down to about 50 ppb for the enzyme in solution. The pesticide detection limit of immobilized enzyme is currently being investigated. The enzyme is capable of a number of measurement cycles without significant loss of signal level.« less

  11. Alkaline pH sensor molecules.

    PubMed

    Murayama, Takashi; Maruyama, Ichiro N

    2015-11-01

    Animals can survive only within a narrow pH range. This requires continual monitoring of environmental and body-fluid pH. Although a variety of acidic pH sensor molecules have been reported, alkaline pH sensor function is not well understood. This Review describes neuronal alkaline pH sensors, grouped according to whether they monitor extracellular or intracellular alkaline pH. Extracellular sensors include the receptor-type guanylyl cyclase, the insulin receptor-related receptor, ligand-gated Cl- channels, connexin hemichannels, two-pore-domain K+ channels, and transient receptor potential (TRP) channels. Intracellular sensors include TRP channels and gap junction channels. Identification of molecular mechanisms underlying alkaline pH sensing is crucial for understanding how animals respond to environmental alkaline pH and how body-fluid pH is maintained within a narrow range. © 2015 Wiley Periodicals, Inc.

  12. Isotopic composition of sulfate accumulations, Northern Calcareous Alps, Austria

    NASA Astrophysics Data System (ADS)

    Bojar, Ana-Voica; Halas, Stanislaw; Bojar, Hans-Peter; Trembaczowski, Andrzej

    2015-04-01

    The Eastern Alps are characterised by the presence of three main tectonic units, such as the Lower, Middle and Upper Austroalpine, which overlie the Penninicum (Tollmann, 1977). The Upper Austroalpine unit consists of the Northern Calcareous Alps (NCA) overlying the Greywacke zone and corresponding to the Graz Paleozoic, Murau Paleozoic and the Gurktal Nappe. Evaporitic rocks are lacking in the later ones. The Northern Calcareous Alps are a detached fold and thrust belt. The sedimentation started in the Late Carboniferous or Early Permian, the age of the youngest sediments being Eocene. The NCA are divided into the Bajuvaric, Tirolic and Juvavic nappe complexes. The evaporitic Haselgebirge Formation occurs in connection with the Juvavic nappe complex at the base of the Tirolic units (Leitner et al., 2013). The Haselgebirge Formation consists mainly of salt, shales, gypsum and anhydrite and includes the oldest sediments of the NCA. The age of the Haselgebirge Formation, established by using spors and geochronological data, is Permian to Lower Triassic. For the Northern Calcareous Alps, the mineralogy of sulphate accumulations consists mainly of gypsum and anhydrite and subordonates of carbonates. The carbonates as magnesite, dolomite and calcite can be found either as singular crystals or as small accumulations within the hosting gypsum. Sulfides (sphalerite, galena, pyrite), sulfarsenides (enargite, baumhauerite) and native sulphur enrichments are known from several deposits (Kirchner, 1987; Postl, 1990). The investigated samples were selected from various gypsum and halite rich deposits of the Northern Calcareous Alps. A total of over 20 samples were investigated, and both oxygen and sulfur isotopic composition were determined for anhydrite, gyps, polyhalite, blödite and langbeinite. The sulfur isotopic values vary between 10.1 to 14 ‰ (CDT), with three values higher than 14 ‰. The Oxygen isotopic values show a range from 9 to 23 ‰ (SMOW). The sulfur

  13. Alkaline battery operational methodology

    DOEpatents

    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.

  14. Low temperature thermochronology in the Eastern Alps: Implications for structural and topographic evolution

    PubMed Central

    Wölfler, Andreas; Stüwe, Kurt; Danišík, Martin; Evans, Noreen J.

    2012-01-01

    According to new apatite fission track, zircon- and apatite (U–Th)/He data, we constrain the near-surface history of the southeastern Tauern Window and adjacent Austrolapine units. The multi-system thermochronological data demonstrate that age-elevation correlations may lead to false implications about exhumation and cooling in the upper crust. We suggest that isothermal warping in the Penninic units that are in the position of a footwall, is due to uplift, erosion and the buildup of topography. Additionally we propose that exhumation rates in the Penninic units did not increase during the Middle Miocene, thus during the time of lateral extrusion. In contrast, exhumation rates of the Austroalpine hangingwall did increase from the Paleogene to the Neogene and the isotherms in this unit were not warped. The new zircon (U–Th)/He ages as well as zircon fission track ages from the literature document a Middle Miocene exhumation pulse which correlates with a period of enhanced sediment accumulation during that time. However, enhanced sedimentation- and exhumation rates at the Miocene/Pliocene boundary, as observed in the Western- and Central Alps, cannot be observed in the Eastern Alps. This contradicts a climatic trigger for surface uplift, and makes a tectonic trigger and/or deep-seated mechanism more obvious to explain surface uplift in the Eastern Alps. In combination with already published geochronological ages, our new data demonstrate Oligocene to Late Miocene fault activity along the Möll valley fault that constitutes a major shear zone in the Eastern Alps. In this context we suggest a geometrical and temporal relationship of the Katschberg-, Polinik–Möll valley- and Mur–Mürz faults that define the extruding wedge in the eastern part of the Eastern Alps. Equal deformation- and fission track cooling ages along the Katschberg–Brenner- and Simplon normal faults demonstrate overall Middle Miocene extension in the whole alpine arc. PMID:27065501

  15. Magnesium ions facilitate integrin alpha 2- and alpha 3-mediated proliferation and enhance alkaline phosphatase expression and activity in hBMSCs.

    PubMed

    Leem, Yea-Hyun; Lee, Kang-Sik; Kim, Jung-Hwa; Seok, Hyun-Kwang; Chang, Jae-Suk; Lee, Dong-Ho

    2016-10-01

    Magnesium metal and its alloys have been proposed as a novel class of bone implant biomaterials because of their biodegradability and mechanical properties. The purpose of this study was to determine whether magnesium ions, which are released abundantly from alloys, affect proliferation and differentiation of human bone marrow-derived stromal cells (hBMSCs). High levels of magnesium ions did not induce cytotoxicity in hBMSCs, but treatment with 2.5-10 mm magnesium ions for 48-72 h significantly increased hBMSC proliferation. The expression of integrins α2 and α3, but not β1, was upregulated compared with the control and shifted from α3 to α2 in hBMSCs treated with magnesium ions. Knockdown of integrins α2 and/or α3 significantly reduced magnesium-induced proliferation of hBMSCs. Magnesium exposure profoundly enhanced alkaline phosphatase (ALP) gene expression and activity even at a relatively low magnesium concentration (2.5 mm). Exposure to magnesium ions facilitated hBMSC proliferation via integrin α2 and α3 expression and partly promoted differentiation into osteoblasts via the alteration of ALP expression and activity. Accordingly, magnesium could be a useful biomaterial for orthopaedic applications such as bone implant biomaterials for repair and regeneration of bone defects in orthopaedic and dental fields. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

  16. Ivrea mantle wedge and arc of the Western Alps (I): Geophysical evidence for the deep structure

    NASA Astrophysics Data System (ADS)

    Kissling, Edi; Schmid, Stefan M.; Diehl, Tobias

    2017-04-01

    The construction of five crustal-scale profiles across the Western Alps and the Ivrea mantle wedge integrates up-to-date geological and geophysical information and reveals important along strike changes in the overall structure of the crust of the Western Alpine arc (Schmid et al. 2017). The 3D crustal model of the Western Alps represented by these cross sections is based on recent P-velocity local earthquake tomography that compliments the previously existing wealth of geophysical information about lithosphere structure in the region. As part of Adria mantle lithosphere exhibiting strong upward bending toward the plate boundary along the inner arc of the Western Alps, the well-known Ivrea body plays a crucial role in our tectonic model. Until recently, however, the detailed 3D geometry of this key structure was only poorly constrained. In this study we present a review of the many seismic data in the region and we document the construction of our 3D lithosphere model by principles of multidisciplinary seismic tomography. Reference: Stefan M. Schmid, Edi Kissling, Douwe J.J. van Hinsbergen, Giancarlo Molli (2017). Ivrea mantle wedge and arc of the Western Alps (2): Kinematic evolution of the Alps-Apennines orogenic system. Abstract Volume EGU 2017.

  17. Holocene geological records of flood regime in French Alps

    NASA Astrophysics Data System (ADS)

    Arnaud, Fabien; Wilhelm, Bruno; Giguet-Covex, Charline; Jenny, Jean-Philippe; Fouinat, Laurent; Sabatier, Pierre; Debret, Maxime; Révillon, Sidonie; Chapron, Emmanuel; Revel, Marie

    2014-05-01

    In this paper we present a review of a ca. 10-years research effort (1-9) aiming at reconstructing floods dynamics in in French Alps through the Holocene, based on lake sediment records. We will particularly discuss how such geological records can be considered as representative of past climate. This implies a wise interpretation of data in order to really understand "what does the core really says". Namely, we showed that different lake systems record different types of flood events. Low altitude lakes, fed by large-scale catchment areas are more sensitive to regional heavy rainfall events (2-5), whereas high altitude small lakes record local extreme rainfall events (6). Moreover, human societies' development must be taken into account as it is susceptible to modulate the climate-geological record relationship (7). Altogether our data permit the establishment of a Holocene-long perspective upon both regional heavy rainfall and torrential activities in high elevation sites. We hence show that both types of events frequency co-evolve in Northern as well as Southern French Alps where Holocene colder spells generally present higher flood frequencies (6-9). On the other hand, intensities of torrential events present a North-South opposite pattern: during warm spells (e.g. the Medieval Warm Period or nowadays), northern Alps are subject to rare but extremely intense heavy rainfall events, whereas in the southern Alps torrential floods are both rare and weak. During cold spells (e.g. the Little Ice Age), the inverse pattern is observed: torrential floods are more frequent everywhere and above-average intensity in Southern Alps. This point is particularly important for risk management in mountain areas in a context of global warming. Our results point out how complex can be the response of regional system to global climate changes. We are hence far from completely understanding this complexity which is moreover imperfectly simulated by climate models. As geological

  18. Alkaline phosphatase and OCT-3/4 as useful markers for predicting susceptibility of human deciduous teeth-derived dental pulp cells to reprogramming factor-induced iPS cells.

    PubMed

    Inada, Emi; Saitoh, Issei; Kubota, Naoko; Soda, Miki; Matsueda, Kazunari; Murakami, Tomoya; Sawami, Tadashi; Kagoshima, Akiko; Yamasaki, Youichi; Sato, Masahiro

    2017-11-01

    The aim of the present study was to prove that primary cells enriched with stem cells are more easily reprogrammed to generate induced pluripotent stem (iPS) cells than those with scarce numbers of stem cells. We surveyed the alkaline phosphatase (ALP) activity in five primarily-isolated human deciduous teeth-derived dental pulp cells (HDDPC) with cytochemical staining to examine the possible presence of stem cells. Next, the expression of stemness-specific factors, such as OCT(Octumer-binding transcription factor)3/4, NANOG, SOX2(SRY (sex determining region Y)-box 2), CD90, muscle segment homeodomain homeobox (MSX) 1, and MSX2, was assessed with a reverse transcription polymerase chain reaction method. Finally, these isolated HDDPC were transfected with plasmids carrying genes coding Yamanaka factors to determine whether these cells could be reprogrammed to generate iPS cells. Of the five primarily-isolated HDDPC, two (HDDPC-1 and -5) exhibited higher degrees of ALP activity. OCT-3/4 expression was also prominent in those two lines. Furthermore, these two lines proliferated faster than the other three lines. The transfection of HDDPC with Yamanaka factors resulted in the generation of iPS cells from HDDPC-1 and -5. The number of cells with the stemness property of HDDPC differs among individuals, which suggests that HDDPC showing an increased expression of both ALP and OCT-3/4 can be more easily reprogrammed to generate iPS cells after the forced expression of reprogramming factors. © 2016 John Wiley & Sons Australia, Ltd.

  19. Phosphate Solubilization and Gene Expression of Phosphate-Solubilizing Bacterium Burkholderia multivorans WS-FJ9 under Different Levels of Soluble Phosphate.

    PubMed

    Zeng, Qingwei; Wu, Xiaoqin; Wang, Jiangchuan; Ding, Xiaolei

    2017-04-28

    Phosphate-solubilizing bacteria (PSB) have the ability to dissolve insoluble phosphate and enhance soil fertility. However, the growth and mineral phosphate solubilization of PSB could be affected by exogenous soluble phosphate and the mechanism has not been fully understood. In the present study, the growth and mineral phosphate-solubilizing characteristics of PSB strain Burkholderia multivorans WS-FJ9 were investigated at six levels of exogenous soluble phosphate (0, 0.5, 1, 5, 10, and 20 mM). The WS-FJ9 strain showed better growth at high levels of soluble phosphate. The phosphate-solubilizing activity of WS-FJ9 was reduced as the soluble phosphate concentration increased, as well as the production of pyruvic acid. Transcriptome profiling of WS-FJ9 at three levels of exogenous soluble phosphate (0, 5, and 20 mM) identified 446 differentially expressed genes, among which 44 genes were continuously up-regulated when soluble phosphate concentration was increased and 81 genes were continuously down-regulated. Some genes related to cell growth were continuously up-regulated, which would account for the better growth of WS-FJ9 at high levels of soluble phosphate. Genes involved in glucose metabolism, including glycerate kinase, 2-oxoglutarate dehydrogenase, and sugar ABC-type transporter, were continuously down-regulated, which indicates that metabolic channeling of glucose towards the phosphorylative pathway was negatively regulated by soluble phosphate. These findings represent an important first step in understanding the molecular mechanisms of soluble phosphate effects on the growth and mineral phosphate solubilization of PSB.

  20. Alginate-hydroxypropylcellulose hydrogel microbeads for alkaline phosphatase encapsulation.

    PubMed

    Karewicz, A; Zasada, K; Bielska, D; Douglas, T E L; Jansen, J A; Leeuwenburgh, S C G; Nowakowska, M

    2014-01-01

    There is a growing interest in using proteins as therapeutics agents. Unfortunately, they suffer from limited stability and bioavailability. We aimed to develop a new delivery system for proteins. ALP, a model protein, was successfully encapsulated in the physically cross-linked sodium alginate/hydroxypropylcellulose (ALG-HPC) hydrogel microparticles. The obtained objects had regular, spherical shape and a diameter of ∼4 µm, as confirmed by optical microscopy and SEM analysis. The properties of the obtained microbeads could be controlled by temperature and additional coating or crosslinking procedures. The slow, sustained release of ALP in its active form with no initial burst effect was observed for chitosan-coated microspheres at pH = 7.4 and 37 °C. Activity of ALP released from ALG/HPC microspheres was confirmed by the occurance of effectively induced mineralization. SEM and AFM images revealed formation of the interpenetrated three-dimensional network of mineral, originating from the microbeads' surfaces. FTIR and XRD analyses confirmed formation of hydroxyapatite.

  1. Differentiating phosphate-dependent and phosphate-independent systemic phosphate-starvation response networks in Arabidopsis thaliana through the application of phosphite

    PubMed Central

    Jost, Ricarda; Pharmawati, Made; Lapis-Gaza, Hazel R.; Rossig, Claudia; Berkowitz, Oliver; Lambers, Hans; Finnegan, Patrick M.

    2015-01-01

    Phosphite is a less oxidized form of phosphorus than phosphate. Phosphite is considered to be taken up by the plant through phosphate transporters. It can mimic phosphate to some extent, but it is not metabolized into organophosphates. Phosphite could therefore interfere with phosphorus signalling networks. Typical physiological and transcriptional responses to low phosphate availability were investigated and the short-term kinetics of their reversion by phosphite, compared with phosphate, were determined in both roots and shoots of Arabidopsis thaliana. Phosphite treatment resulted in a strong growth arrest. It mimicked phosphate in causing a reduction in leaf anthocyanins and in the expression of a subset of the phosphate-starvation-responsive genes. However, the kinetics of the response were slower than for phosphate, which may be due to discrimination against phosphite by phosphate transporters PHT1;8 and PHT1;9 causing delayed shoot accumulation of phosphite. Transcripts encoding PHT1;7, lipid-remodelling enzymes such as SQD2, and phosphocholine-producing NMT3 were highly responsive to phosphite, suggesting their regulation by a direct phosphate-sensing network. Genes encoding components associated with the ‘PHO regulon’ in plants, such as At4, IPS1, and PHO1;H1, generally responded more slowly to phosphite than to phosphate, except for SPX1 in roots and MIR399d in shoots. Two uncharacterized phosphate-responsive E3 ligase genes, PUB35 and C3HC4, were also highly phosphite responsive. These results show that phosphite is a valuable tool to identify network components directly responsive to phosphate. PMID:25697796

  2. Carbonatite and alkaline intrusion-related rare earth element deposits–A deposit model

    USGS Publications Warehouse

    Verplanck, Philip L.; Van Gosen, Bradley S.

    2011-01-01

    The rare earth elements are not as rare in nature as their name implies, but economic deposits with these elements are not common and few deposits have been large producers. In the past 25 years, demand for rare earth elements has increased dramatically because of their wide and diverse use in high-technology applications. Yet, presently the global production and supply of rare earth elements come from only a few sources. China produces more than 95 percent of the world's supply of rare earth elements. Because of China's decision to restrict exports of these elements, the price of rare earth elements has increased and industrial countries are concerned about supply shortages. As a result, understanding the distribution and origin of rare earth elements deposits, and identifying and quantifying our nation's rare earth elements resources have become priorities. Carbonatite and alkaline intrusive complexes, as well as their weathering products, are the primary sources of rare earth elements. The general mineral deposit model summarized here is part of an effort by the U.S. Geological Survey's Mineral Resources Program to update existing models and develop new descriptive mineral deposit models to supplement previously published models for use in mineral-resource and mineral-environmental assessments. Carbonatite and alkaline intrusion-related REE deposits are discussed together because of their spatial association, common enrichment in incompatible elements, and similarities in genesis. A wide variety of commodities have been exploited from carbonatites and alkaline igneous rocks, such as rare earth elements, niobium, phosphate, titanium, vermiculite, barite, fluorite, copper, calcite, and zirconium. Other enrichments include manganese, strontium, tantalum, thorium, vanadium, and uranium.

  3. Anodes for alkaline electrolysis

    DOEpatents

    Soloveichik, Grigorii Lev [Latham, NY

    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.

  4. Continuous micro-earthquake catalogue of the central Southern Alps, New Zealand

    NASA Astrophysics Data System (ADS)

    Michailos, Konstantinos; Townend, John; Savage, Martha; Chamberlain, Calum

    2017-04-01

    The Alpine Fault is one of the most prominent tectonic features in the South Island, New Zealand, and is inferred to be late in its seismic cycle of M 8 earthquakes based on paleoseismological evidence. Despite this, the Alpine Fault displays low levels of contemporary seismic activity, with little documented on-fault seismicity. This low magnitude seismicity, often below the completeness level of the GeoNet national seismic catalogue, may inform us of changes in fault character along-strike and might be used for rupture simulations and hazard planning. Thus, compiling a micro-earthquake catalogue for the Southern Alps prior to an expected major earthquake is of great interest. Areas of low seismic activity, like the central part of the Alpine Fault, require data recorded over a long duration to reveal temporal and spatial seismicity patterns and provide a better understanding for the processes controlling seismogenesis. The continuity and density of the Southern Alps Microearthquake Borehole Array (SAMBA; deployed in late 2008) allows us to study seismicity in the Southern Alps over a more extended time period than has ever been done previously. Furthermore, by using data from other temporary networks (e.g. WIZARD, ALFA08, DFDP-10) we are able to extend the region covered. To generate a spatially and temporally continuous catalogue of seismicity in New Zealand's central Southern Alps, we used automatic detection and phase-picking methods. We used an automatic phase-picking method for both P- and S- wave arrivals (kPick; Rawles and Thurber, 2015). Using almost 8 years of seismic data we calculated about 9,000 preliminary earthquake. The seismicity is clustered and scattered and a previously observed seismic gap between the Wanganui and Whataroa rivers is also identified.

  5. Bergsteigen in den Alpen (Mountain Climbing in the Alps).

    ERIC Educational Resources Information Center

    Hawrysz, Ilse; Budzinski, Elisabeth

    German second language instructional materials contain a short text in German on mountain climbing in the Alps, a vocabulary list with translation, a simple German climbing song, a recipe for goulash soup in English, and a short text in English on mountain climbing. (MSE)

  6. Effect of a Particulate and a Putty-Like Tricalcium Phosphate-Based Bone-grafting Material on Bone Formation, Volume Stability and Osteogenic Marker Expression after Bilateral Sinus Floor Augmentation in Humans

    PubMed Central

    Knabe, Christine; Adel Khattab, Doaa; Kluk, Esther; Struck, Rainer; Stiller, Michael

    2017-01-01

    This study examines the effect of a hyaluronic acid (HyAc) containing tricalcium phosphate putty scaffold material (TCP-P) and of a particulate tricalcium phosphate (TCP-G) graft on bone formation, volume stability and osteogenic marker expression in biopsies sampled 6 months after bilateral sinus floor augmentation (SFA) in 7 patients applying a split-mouth design. 10% autogenous bone chips were added to the grafting material during surgery. The grain size of the TCP granules was 700 to 1400 µm for TCP-G and 125 to 250 µm and 500 to 700 µm (ratio 1:1) for TCP-P. Biopsies were processed for immunohistochemical analysis of resin-embedded sections. Sections were stained for collagen type I (Col I), alkaline phosphatase (ALP), osteocalcin (OC) and bone sialoprotein (BSP). Furthermore, the bone area and biomaterial area fraction were determined histomorphometrically. Cone-beam CT data recorded after SFA and 6 months later were used for calculating the graft volume at these two time points. TCP-P displayed more advantageous surgical handling properties and a significantly greater bone area fraction and smaller biomaterial area fraction. This was accompanied by significantly greater expression of Col I and BSP and in osteoblasts and osteoid and a less pronounced reduction in grafting volume with TCP-P. SFA using both types of materials resulted in formation of sufficient bone volume for facilitating stable dental implant placement with all dental implants having been in function without any complications for 6 years. Since TCP-P displayed superior surgical handling properties and greater bone formation than TCP-G, without the HyAc hydrogel matrix having any adverse effect on bone formation or graft volume stability, TCP-P can be regarded as excellent grafting material for SFA in a clinical setting. The greater bone formation observed with TCP-P may be related to the difference in grain size of the TCP granules and/or the addition of the HyAc. PMID:28758916

  7. Surfactant-enhanced alkaline flooding: Buffering at intermediate alkaline pH

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rudin, J.; Wasan, D.T.

    1993-11-01

    The alkaline flooding process involves injecting alkaline agents into the reservoir to produce more oil than is produced through conventional waterflooding. The interaction of the alkali in the flood water with the naturally occurring acids in the reservoir oil results in in-situ formation of soaps, which are partially responsible for lowering IFT and improving oil recovery. The extent to which IFT is lowered depends on the specific oil and injection water properties. Numerous investigators have attempted to clarify the relationship between system chemical composition and IFT. An experimental investigation of buffered alkaline flooding system chemistry was undertaken to determine themore » influence of various species present on interfacial tension (IFT) as a function of pH and ionic strength. IFT was found to go through an ultralow minimum in certain pH ranges. This synergism results from simultaneous adsorption of un-ionized and ionized acid species on the interface.« less

  8. Enzyme Replacement Therapy for Murine Hypophosphatasia*

    PubMed Central

    Millán, José Luis; Narisawa, Sonoko; Lemire, Isabelle; Loisel, Thomas P; Boileau, Guy; Leonard, Pierre; Gramatikova, Svetlana; Terkeltaub, Robert; Camacho, Nancy Pleshko; McKee, Marc D; Crine, Philippe; Whyte, Michael P

    2008-01-01

    Introduction Hypophosphatasia (HPP) is the inborn error of metabolism that features rickets or osteomalacia caused by loss-of-function mutation(s) within the gene that encodes the tissue-nonspecific isozyme of alkaline phosphatase (TNALP). Consequently, natural substrates for this ectoenzyme accumulate extracellulary including inorganic pyrophosphate (PPi), an inhibitor of mineralization, and pyridoxal 5`-phosphate (PLP), a co-factor form of vitamin B6. Babies with the infantile form of HPP often die with severe rickets and sometimes hypercalcemia and vitamin B6-dependent seizures. There is no established medical treatment. Materials and Methods Human TNALP was bioengineered with the C terminus extended by the Fc region of human IgG for one-step purification and a deca-aspartate sequence (D10) for targeting to mineralizing tissue (sALP-FcD10). TNALP-null mice (Akp2−/−), an excellent model for infantile HPP, were treated from birth using sALP-FcD10. Short-term and long-term efficacy studies consisted of once daily subcutaneous injections of 1, 2, or 8.2 mg/kg sALP-FcD10 for 15, 19, and 15 or 52 days, respectively. We assessed survival and growth rates, circulating levels of sALP-FcD10 activity, calcium, PPi, and pyridoxal, as well as skeletal and dental manifestations using radiography, μCT, and histomorphometry. Results Akp2−/− mice receiving high-dose sALP-FcD10 grew normally and appeared well without skeletal or dental disease or epilepsy. Plasma calcium, PPi, and pyridoxal concentrations remained in their normal ranges. We found no evidence of significant skeletal or dental disease. Conclusions Enzyme replacement using a bone-targeted, recombinant form of human TNALP prevents infantile HPP in Akp2−/− mice. PMID:18086009

  9. Electrochemical Branched-DNA Assay for Polymerase Chain Reaction-Free Detection and Quantification of Oncogenes in Messenger RNA

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lee, Ai Cheng; Dai, Ziyu; Chen, Baowei

    2008-12-01

    We describe a novel electrochemical branched-DNA (bDNA) assay for polymerase chain reaction (PCR)-free detection and quantification of p185 BCR-ABL leukemia fusion transcript in the population of messenger RNA (mRNA) extracted from cell lines. The bDNA amplifier carrying high loading of alkaline phosphatase (ALP) tracers was used to amplify targets signal. The targets were captured on microplate well surfaces through cooperative sandwich hybridization prior to the labeling of bDNA. The activity of captured ALP was monitored by square-wave voltammetric (SWV) analysis of the electroactive enzymatic product in the presence of 1-napthyl-phosphate. The specificity and sensitivity of assay enabled direct detection ofmore » target transcript in as little as 4.6 ng mRNA without PCR amplification. In combination with the use of a well-quantified standard, the electrochemical bDNA assay was capable of direct use for a PCR-free quantitative analysis of target transcript in total mRNA population. The approach thus provides a simple, sensitive, accurate and quantitative tool alternate to the RQ-PCR for early disease diagnosis.« less

  10. Microwave assisted preparation of magnesium phosphate cement (MPC) for orthopedic applications: a novel solution to the exothermicity problem.

    PubMed

    Zhou, Huan; Agarwal, Anand K; Goel, Vijay K; Bhaduri, Sarit B

    2013-10-01

    There are two interesting features of this paper. First, we report herein a novel microwave assisted technique to prepare phosphate based orthopedic cements, which do not generate any exothermicity during setting. The exothermic reactions during the setting of phosphate cements can cause tissue damage during the administration of injectable compositions and hence a solution to the problem is sought via microwave processing. This solution through microwave exposure is based on a phenomenon that microwave irradiation can remove all water molecules from the alkaline earth phosphate cement paste to temporarily stop the setting reaction while preserving the active precursor phase in the formulation. The setting reaction can be initiated a second time by adding aqueous medium, but without any exothermicity. Second, a special emphasis is placed on using this technique to synthesize magnesium phosphate cements for orthopedic applications with their enhanced mechanical properties and possible uses as drug and protein delivery vehicles. The as-synthesized cements were evaluated for the occurrences of exothermic reactions, setting times, presence of Mg-phosphate phases, compressive strength levels, microstructural features before and after soaking in (simulated body fluid) SBF, and in vitro cytocompatibility responses. The major results show that exposure to microwaves solves the exothermicity problem, while simultaneously improving the mechanical performance of hardened cements and reducing the setting times. As expected, the cements are also found to be cytocompatible. Finally, it is observed that this process can be applied to calcium phosphate cements system (CPCs) as well. Based on the results, this microwave exposure provides a novel technique for the processing of injectable phosphate bone cement compositions. © 2013.

  11. Hypogeous fungi at tree line in the Australian Alps

    Treesearch

    James M. Trappe; Andrew W Claridge

    2006-01-01

    The tree line of the continental Australian Alps yielded eighteen species of hypogeous fungi, all probably . forming ectomycorrhizae with Eucalyptus niphophila, the tree species characteristic of that habitat. Six of the species were undescribed. These collections represented six families and twelve genera: Boletaceae (with Chamonixia), Gallaceaceae...

  12. Preliminary research on a novel bioactive silicon doped calcium phosphate coating on AZ31 magnesium alloy via electrodeposition.

    PubMed

    Qiu, Xun; Wan, Peng; Tan, Lili; Fan, Xinmin; Yang, Ke

    2014-03-01

    A silicon doped calcium phosphate coating was obtained successfully on AZ31 alloy substrate via pulse electrodeposition. A novel dual-layer structure was observed with a porous lamellar-like and outer block-like apatite layer. In vitro immersion tests were adopted in simulated body fluid within 28 days of immersion. Slow degradation rate obtained from weight loss was observed for the Si-doped Ca-P coating, which was also consistent with the results of electrochemical experiments showing an enhanced corrosion resistance for the coating. Further formation of an apatite-like layer on the surface after immersion proved better integrity and biomineralization performance of the coating. Biological characterization was carried out for viability, proliferation and differentiation of MG63 osteoblast-like cells. The coating showed a good cell growth and an enhanced cell proliferation. Moreover, an increased activity of osteogenic marker ALP was found. All the results demonstrated that the Si-doped calcium phosphate was perspective to be used as a coating for magnesium alloy implants to control the degradation rate and enhance the bioactivity, which would facilitate the rapidity of bone tissue repair. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Effect of alkaline addition on anaerobic sludge digestion with combined pretreatment of alkaline and high pressure homogenization.

    PubMed

    Fang, Wei; Zhang, Panyue; Zhang, Guangming; Jin, Shuguang; Li, Dongyi; Zhang, Meixia; Xu, Xiangzhe

    2014-09-01

    To improve anaerobic digestion efficiency, combination pretreatment of alkaline and high pressure homogenization was applied to pretreat sewage sludge. Effect of alkaline dosage on anaerobic sludge digestion was investigated in detail. SCOD of sludge supernatant significantly increased with the alkaline dosage increase after the combined pretreatment because of sludge disintegration. Organics were significantly degraded after the anaerobic digestion, and the maximal SCOD, TCOD and VS removal was 73.5%, 61.3% and 43.5%, respectively. Cumulative biogas production, methane content in biogas and biogas production rate obviously increased with the alkaline dosage increase. Considering both the biogas production and alkaline dosage, the optimal alkaline dosage was selected as 0.04 mol/L. Relationships between biogas production and sludge disintegration showed that the accumulative biogas was mainly enhanced by the sludge disintegration. The methane yield linearly increased with the DDCOD increase as Methane yield (ml/gVS)=4.66 DDCOD-9.69. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. A Hannibal's Treck Across The Alps: Geomorphological Analysis Of Sites Of Geoarchaeologicals Interest

    NASA Astrophysics Data System (ADS)

    Mahaney, William C.; Kalm, Volli; Dirszowsky, Randy W.; Milner, Michael W.; Sodhi, Rana; Beukens, Roelf; Dorn, Ron; Tricart, Pierre; Schwartz, Stéphane; Chamorro-Perez, Eva; Boccia, Sal; Barendregt, René W.; Krinsley, D. H.; Seaquist, E. R.; Merrick, David; Kapran, Barbara

    A ~2200 year-old question related to Hannibal's invasion route across the Alps into Italia, has been argued by classicists without recovery of material evidence. A comparison of topographical descriptions in the ancient literature with environmental parameters in the Alps, attempted here for the first time, provides a database against which various pathways can be assessed. Identification of sites using geological, geomorphological, astronomical, chemical and petrological methods leads to the exclusion of certain transit points and targeting of others where geoarchaeological excavation might yield important evidence related to the military culture of ancient Carthage.

  15. Pediatric reference intervals for alkaline phosphatase.

    PubMed

    Zierk, Jakob; Arzideh, Farhad; Haeckel, Rainer; Cario, Holger; Frühwald, Michael C; Groß, Hans-Jürgen; Gscheidmeier, Thomas; Hoffmann, Reinhard; Krebs, Alexander; Lichtinghagen, Ralf; Neumann, Michael; Ruf, Hans-Georg; Steigerwald, Udo; Streichert, Thomas; Rascher, Wolfgang; Metzler, Markus; Rauh, Manfred

    2017-01-01

    Interpretation of alkaline phosphatase activity in children is challenging due to extensive changes with growth and puberty leading to distinct sex- and age-specific dynamics. Continuous percentile charts from birth to adulthood allow accurate consideration of these dynamics and seem reasonable for an analyte as closely linked to growth as alkaline phosphatase. However, the ethical and practical challenges unique to pediatric reference intervals have restricted the creation of such percentile charts, resulting in limitations when clinical decisions are based on alkaline phosphatase activity. We applied an indirect method to generate percentile charts for alkaline phosphatase activity using clinical laboratory data collected during the clinical care of patients. A total of 361,405 samples from 124,440 patients from six German tertiary care centers and one German laboratory service provider measured between January 2004 and June 2015 were analyzed. Measurement of alkaline phosphatase activity was performed on Roche Cobas analyzers using the IFCC's photometric method. We created percentile charts for alkaline phosphatase activity in girls and boys from birth to 18 years which can be used as reference intervals. Additionally, data tables of age- and sex-specific percentile values allow the incorporation of these results into laboratory information systems. The percentile charts provided enable the appropriate differential diagnosis of changes in alkaline phosphatase activity due to disease and changes due to physiological development. After local validation, integration of the provided percentile charts into result reporting facilitates precise assessment of alkaline phosphatase dynamics in pediatrics.

  16. Phosphates at the Surface of Mars: Primary Deposits and Alteration Products

    NASA Technical Reports Server (NTRS)

    Yen, Albert S.; Gellert, Ralf; Clark, Benton C.; Ming, Douglas W.; Mittlefehldt, David W.; Arvidson, Raymond E.; McSween, Harry Y., Jr.; Schroder, Christian

    2014-01-01

    Phosphorus is an essential element in terrestrial organisms and thus characterizing the occurrences of phosphate phases at the martian surface is crucial in the assessment of habitability. The Alpha Particle X-Ray Spectrometers onboard Spirit, Opportunity and Curiosity discovered a variety of primary and secondary phosphate phases allowing direct comparisons across the three landing sites. The Spirit rover at Gusev Crater encountered the "Wishstone/Watchtower" class of P-rich (up to 5.2 wt% P2O5) rocks interpreted to be alkaline volcanic rocks with a physical admixture of approximately 10 to 20% merrillite [Usui et al 2008]. These rocks are characterized by elevated Ti and Y and anomalously low Cr and Ni, which could largely reflect the nature of the protoliths: Evolved magmatic rocks. Many of these chemical signatures are also found in pyroclastic deposits at nearby "Home Plate" and in phosphate precipitates derived from fluid interactions with these rocks ("Paso Robles" soils). The Opportunity rover at Meridiani Planum recently analyzed approximately 4 cm clast in a fine-grained matrix, one of numerous rocks of similar appearance at the rim of Endeavour Crater. This clast, "Sarcobatus," has minor enrichments in Ca and P relative to the matrix, and like the P-rich rocks at Gusev, Sarcobatus also shows elevated Al and Ti. On the same segment of the Endeavour rim, subsurface samples were found with exceptional levels of Mn (approximately 3.5 wt% MnO). These secondary and likely aqueous deposits contain strong evidence for associated Mg-sulfate and Ca-phosphate phases. Finally, the Curiosity traverse at Gale crater encountered P-rich rocks compositionally comparable to Wishstone at Gusev, including elevated Y. Phosphorous-rich rocks with similar chemical characteristics are prevalent on Mars, and the trace and minor element signatures provide constraints on whether these are primary deposits, secondary products of physical weathering or secondary products of chemical

  17. Bioavailable dietary phosphate, a mediator of cardiovascular disease, may be decreased with plant-based diets, phosphate binders, niacin, and avoidance of phosphate additives.

    PubMed

    McCarty, Mark F; DiNicolantonio, James J

    2014-01-01

    Increased fasting serum phosphate within the normal physiological range has been linked to increased cardiovascular risk in prospective epidemiology; increased production of fibroblast growth factor 23, and direct vascular effects of phosphate, may mediate this risk. Although dietary phosphate intake does not clearly influence fasting serum phosphate in individuals with normal renal function, increased phosphate intake can provoke a rise in fibroblast growth factor 23, and in diurnal phosphate levels, and hence may adversely influence vascular health. Dietary phosphate absorption can be moderated by emphasizing plant-based dietary choices (which provide phosphate in less bioavailable forms); avoidance of processed foods containing inorganic phosphate food additives; and by ingestion of phosphate-binder drugs, magnesium supplements, or niacin, which precipitate phosphate or suppress its gastrointestinal absorption. The propensity of dietary phosphate to promote vascular calcification may be opposed by optimal intakes of magnesium, vitamin K, and vitamin D; the latter should also counter the tendency of phosphate to elevate parathyroid hormone. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. The Alkaline Diet: Is There Evidence That an Alkaline pH Diet Benefits Health?

    PubMed Central

    Schwalfenberg, Gerry K.

    2012-01-01

    This review looks at the role of an alkaline diet in health. Pubmed was searched looking for articles on pH, potential renal acid loads, bone health, muscle, growth hormone, back pain, vitamin D and chemotherapy. Many books written in the lay literature on the alkaline diet were also reviewed and evaluated in light of the published medical literature. There may be some value in considering an alkaline diet in reducing morbidity and mortality from chronic diseases and further studies are warranted in this area of medicine. PMID:22013455

  19. Thin visous sheet modelling of orogen scale deformation. The Eastern Alps in plan view

    NASA Astrophysics Data System (ADS)

    Robl, J.; Stuewe, K.

    2003-04-01

    We present first results of a new numerical model to describe the dynamic evolution of the eastern Alps in plan view on the orogen scale. We investigate the influence of boundary conditions, gravitational potential energy, rheology contrast of major tectonic units and internal structures on the deformation field. We aim at estimating the Argan number of the Eastern Alps and to calculate slip rates along big lineaments that represent the northern and southern border of the Austro-Alpine extrusion corridor. A further aim is to predict the position and the activity of major faults where they disappear below quartenary sediments. All calculation are perfomed with the mechanical finite element code BASIL that allows computation on a thin visous sheet. The starting conditons are controlled by the varying crustal thickness of the region and by the rheolgy of the Adriatic indenter in the south, the Bohemian massif in the north and the Eastern Alps inbetween. We assume that the Eastern Alps are fixed to the north and the west while the southern boundary moves northward at a rate of 6-8 mm /y. The geodynamic setting in the east changed over the last 5 my. While a roll back subduction zone beneath the Carparthian belt accompanied by extension, crustal thinning and basin formation controlled the deformation of the Eastern Alps until the Miocene. Later on subduction stopped and the overall stress field changed from extension to compression resulting in uplift of many basins and the lack sediments younger than Pliocene. This well known variation of plate tectonic scenarios over the last 5 my allows us to place tight constraints for the boundary conditions of our model. Although our work is only now in progress, we can report some promising results: some of the kinematics and mechaniscs predicted by our modelling are consistent with field observations of the structural geologists and geodeticists.

  20. Ice-borne prehistoric finds in the Swiss Alps reflect Holocene glacier fluctuations

    NASA Astrophysics Data System (ADS)

    Grosjean, Martin; Suter, Peter J.; Trachsel, Mathias; Wanner, Heinz

    2007-03-01

    During the hot summer of 2003, reduction of an ice field in the Swiss Alps (Schnidejoch) uncovered spectacular archaeological hunting gear, fur, leather and woollen clothing and tools from four distinct windows of time: Neolithic Age (4900 to 4450 cal. yr BP), early Bronze Age (4100-3650 cal. yr BP), Roman Age (1st-3rd century AD), and Medieval times (8-9th century AD and 14-15th century AD). Transalpine routes connecting northern Italy with the northern Alps during these slots is consistent with late Holocene maximum glacier retreat. The age cohorts of the artefacts are separated which is indicative of glacier advances when the route was difficult and not used for transit. The preservation of Neolithic leather indicates permanent ice cover at that site from ca. 4900 cal. yr BP until AD 2003, implying that the ice cover was smaller in 2003 than at any time during the last 5000 years. Current glacier retreat is unprecedented since at least that time. This is highly significant regarding the interpretation of the recent warming and the rapid loss of ice in the Alps. Copyright

  1. The Ortles ice cores: uncovering an extended climate archive from the Eastern Alps

    NASA Astrophysics Data System (ADS)

    Dreossi, Giuliano; Barbante, Carlo; Bertò, Michele; Carturan, Luca; De Blasi, Fabrizio; Gabrieli, Jacopo; Gabrielli, Paolo; Seppi, Roberto; Spolaor, Andrea; Stenni, Barbara; Zanoner, Thomas

    2017-04-01

    During the last half century, oxygen and hydrogen stable isotope content of ice cores has been extensively used for air temperature reconstructions. The most suitable glaciers of the Alpine area, most exclusively in the Western Alps, have been utilized for ice coring for more than four decades. The paleoclimatic potential of the Eastern Alps is still largely unexploited and was scarcely utilized in the past mainly because of the lower elevation (compared to Western Alps) and hence the difficulty to find glaciers in cold conditions. The warming temperature trend appears to be particularly pronounced in the Alps, threatening the preservation of the glaciated areas and creating a sense of urgency in retrieving climatic archives before it is too late. In autumn 2011, four deep cores were drilled on Mt Ortles, South Tyrol, Italy, at 3859 m a.s.l. An extensive reconstructed temperature record for the Ortles summit, based on the surrounding meteorological station data, is available for the last 150 years, while an automatic weather station had been operating from 2011 to 2015 in proximity of the drilling site. The new ice core chronology, based on 210Pb, tritium, beta emissions analysis and 14C measurements of the particulate organic carbon, indicates that the bottom ice is 7000 years old, making it the second most extended glaciological archive ever retrieved in the Alps. The three equally long ice cores have been analyzed for oxygen and hydrogen stable isotopes throughout their length, and the goal is to create an Ortles stacked record for d18O and dD and compare the isotopic data to instrumental temperatures and to other Alpine records. Since 2008, several snow pits were dug in proximity of the drilling site during summer, when the temperature can often exceed the melting point. The isotopic profiles of the 2015 snow pit, dug at the end of an exceptionally warm summer, show how the isotope signal is now affected by the post-depositional processes that have occurred

  2. Reconstructing the Alps-Carpathians-Dinarides as a key to understanding switches in subduction polarity, slab gaps and surface motion

    NASA Astrophysics Data System (ADS)

    Handy, Mark R.; Ustaszewski, Kamil; Kissling, Eduard

    2015-01-01

    Palinspastic map reconstructions and plate motion studies reveal that switches in subduction polarity and the opening of slab gaps beneath the Alps and Dinarides were triggered by slab tearing and involved widespread intracrustal and crust-mantle decoupling during Adria-Europe collision. In particular, the switch from south-directed European subduction to north-directed "wrong-way" Adriatic subduction beneath the Eastern Alps was preconditioned by two slab-tearing events that were continuous in Cenozoic time: (1) late Eocene to early Oligocene rupturing of the oppositely dipping European and Adriatic slabs; these ruptures nucleated along a trench-trench transfer fault connecting the Alps and Dinarides; (2) Oligocene to Miocene steepening and tearing of the remaining European slab under the Eastern Alps and western Carpathians, while subduction of European lithosphere continued beneath the Western and Central Alps. Following the first event, post-late Eocene NW motion of the Adriatic Plate with respect to Europe opened a gap along the Alps-Dinarides transfer fault which was filled with upwelling asthenosphere. The resulting thermal erosion of the lithosphere led to the present slab gap beneath the northern Dinarides. This upwelling also weakened the upper plate of the easternmost part of the Alpine orogen and induced widespread crust-mantle decoupling, thus facilitating Pannonian extension and roll-back subduction of the Carpathian oceanic embayment. The second slab-tearing event triggered uplift and peneplainization in the Eastern Alps while opening a second slab gap, still present between the Eastern and Central Alps, that was partly filled by northward counterclockwise subduction of previously unsubducted Adriatic continental lithosphere. In Miocene time, Adriatic subduction thus jumped westward from the Dinarides into the heart of the Alpine orogen, where northward indentation and wedging of Adriatic crust led to rapid exhumation and orogen-parallel escape of

  3. 2-Substituted 7-trifluoromethyl-thiadiazolopyrimidones as alkaline phosphatase inhibitors. Synthesis, structure activity relationship and molecular docking study.

    PubMed

    Jafari, Behzad; Ospanov, Meirambek; Ejaz, Syeda Abida; Yelibayeva, Nazym; Khan, Shafi Ullah; Amjad, Sayyeda Tayyeba; Safarov, Sayfidin; Abilov, Zharylkasyn A; Turmukhanova, Mirgul Zh; Kalugin, Sergey N; Ehlers, Peter; Lecka, Joanna; Sévigny, Jean; Iqbal, Jamshed; Langer, Peter

    2018-01-20

    Alkaline Phosphatases (APs) play a key role in maintaining a ratio of phosphate to inorganic pyrophosphate (P i /PP i ) and thus regulate extracellular matrix calcification during bone formation and growth. Among different isozymes of AP, aberrant increase in the level of tissue non-specific alkaline phosphatase (TNAP) is strongly associated with vascular calcification and end-stage renal diseases. In this context, we synthesized a novel series of fluorinated pyrimidone derivatives, i.e., 2-bromo-7-trifluoromethyl-5-oxo-5H-1,3,4-thiadiazolepyrimidones. The bromine functionality was further used for derivatisation by nucleophilic aromatic substitution using amines as nucleophiles as well as by Palladium catalysed Suzuki-Miyaura reactions. The synthesized derivatives were found potent but non-selective inhibitors of both isozymes of AP. Arylated thiadiazolopyrimidones exhibited stronger inhibitory activities than 2-amino-thiadiazolopyrimidones. The binding modes and possible interactions of the most active inhibitor within the active site of the enzyme were observed by molecular docking studies. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  4. Downscaling Alkaline Phosphatase Activity in a Subtropical Reservoir

    NASA Astrophysics Data System (ADS)

    Tseng, Y.

    2011-12-01

    This research was conducted by downscaling study to understand phosphorus (P)-deficient status of different plankton and the role of alkaline phosphatase activity (APA) in subtropical Feitsui Reservoir. Results from field survey showed that bulk APA (1.6~95.2 nM h-1) was widely observed in the epilimnion (0~20 m) with an apparent seasonal variations, suggesting that plankton in the system were subjected to P-deficient seasonally. Mixed layer depth (an index of phosphate availability) is the major factor influencing the variation of bulk APA and specific APA (124~1,253 nmol mg C-1 h-1), based on multiple linear regression analysis. Size-fractionated APA assays showed that picoplankton (size 0.2~3 um) contributed most of the bulk APA in the system. In addition, single-cell APA detected by enzyme-labeled fluorescence (ELF) assay indicated that heterotrophic bacteria are the major contributors of APA. Thus, we can infer that bacteria play an important role in accelerating P-cycle within P-deficient systems. Light/nutrient manipulation bioassays showed that bacterial growth was directly controlled by phosphate, while picocyanobacterial growth is controlled by light and can out-compete bacteria under P-limited condition with the aid of light. Further analysis revealed that the strength of summer typhoon is a factor responsible for the inter-annual variability of bulk and specific APA. APA study demonstrated the episodic events (e.g. strong typhoon and extreme precipitation) had significant influence on APA variability in sub-tropical to tropical aquatic ecosystems. Hence, the results herein will allow future studies on monitoring typhoon disturbance (intensity and frequency) as well as the APA of plankton during summer-to-autumn in subtropical systems.

  5. Characterization of phosphorus leaching from phosphate waste rock in the Xiangxi River watershed, Three Gorges Reservoir, China.

    PubMed

    Jiang, Li-Guo; Liang, Bing; Xue, Qiang; Yin, Cheng-Wei

    2016-05-01

    Phosphate mining waste rocks dumped in the Xiangxi River (XXR) bay, which is the largest backwater zone of the Three Gorges Reservoir (TGR), are treated as Type I industry solid wastes by the Chinese government. To evaluate the potential pollution risk of phosphorus leaching from phosphate waste rocks, the phosphorus leaching behaviors of six phosphate waste rock samples with different weathering degrees under both neutral and acidic conditions were investigated using a series of column leaching experiments, following the Method 1314 standard of the US EPA. The results indicate that the phosphorus release mechanism is solubility-controlled. Phosphorus release from waste rocks increases as pH decreases. The phosphorus leaching concentration and cumulative phosphorus released in acidic leaching conditions were found to be one order of magnitude greater than that in neutral leaching conditions. In addition, the phosphorus was released faster during the period when environmental pH turned from weak alkalinity to slight acidity, with this accelerated release period appearing when L/S was in the range of 0.5-2.0 mL/g. In both neutral and acidic conditions, the average values of Total Phosphorus (TP), including orthophosphates, polyphosphates and organic phosphate, leaching concentration exceed the availability by regulatory (0.5 mg/L) in the whole L/S range, suggesting that the phosphate waste rocks stacked within the XXR watershed should be considered as Type II industry solid wastes. Therefore, the phosphate waste rocks deposited within the study area should be considered as phosphorus point pollution sources, which could threaten the adjacent surface-water environment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Characterization and quantification of biochar alkalinity.

    PubMed

    Fidel, Rivka B; Laird, David A; Thompson, Michael L; Lawrinenko, Michael

    2017-01-01

    Lack of knowledge regarding the nature of biochar alkalis has hindered understanding of pH-sensitive biochar-soil interactions. Here we investigate the nature of biochar alkalinity and present a cohesive suite of methods for its quantification. Biochars produced from cellulose, corn stover and wood feedstocks had significant low-pK a organic structural (0.03-0.34 meq g -1 ), other organic (0-0.92 meq g -1 ), carbonate (0.02-1.5 meq g -1 ), and other inorganic (0-0.26 meq g -1 ) alkalinities. All four categories of biochar alkalinity contributed to total biochar alkalinity and are therefore relevant to pH-sensitive soil processes. Total biochar alkalinity was strongly correlated with base cation concentration, but biochar alkalinity was not a simple function of elemental composition, soluble ash, fixed carbon, or volatile matter content. More research is needed to characterize soluble biochar alkalis other than carbonates and to establish predictive relationships among biochar production parameters and the composition of biochar alkalis. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Mussel-inspired bioceramics with self-assembled Ca-P/polydopamine composite nanolayer: preparation, formation mechanism, improved cellular bioactivity and osteogenic differentiation of bone marrow stromal cells.

    PubMed

    Wu, Chengtie; Han, Pingping; Liu, Xiaoguo; Xu, Mengchi; Tian, Tian; Chang, Jiang; Xiao, Yin

    2014-01-01

    The nanostructured surface of biomaterials plays an important role in improving their in vitro cellular bioactivity as well as stimulating in vivo tissue regeneration. Inspired by the mussel's adhesive versatility, which is thought to be due to the plaque-substrate interface being rich in 3,4-dihydroxy-l-phenylalamine (DOPA) and lysine amino acids, in this study we developed a self-assembly method to prepare a uniform calcium phosphate (Ca-P)/polydopamine composite nanolayer on the surface of β-tricalcium phosphate (β-TCP) bioceramics by soaking β-TCP bioceramics in Tris-dopamine solution. It was found that the addition of dopamine, reaction temperature and reaction time are three key factors inducing the formation of a uniform Ca-P/polydopamine composite nanolayer. The formation mechanism of a Ca-P/polydopamine composite nanolayer involved two important steps: (i) the addition of dopamine to Tris-HCl solution decreases the pH value and accelerates Ca and P ionic dissolution from the crystal boundaries of β-TCP ceramics; (ii) dopamine is polymerized to form self-assembled polydopamine film and, at the same time, nanosized Ca-P particles are mineralized with the assistance of polydopamine, in which the formation of polydopamine occurs simultaneously with Ca-P mineralization (formation of nanosized microparticles composed of calcium phosphate-based materials), and finally a self-assembled Ca-P/polydopamine composite nanolayer forms on the surface of the β-TCP ceramics. Furthermore, the formed self-assembled Ca-P/polydopamine composite nanolayer significantly enhances the surface roughness and hydrophilicity of β-TCP ceramics, and stimulates the attachment, proliferation, alkaline phosphate (ALP) activity and bone-related gene expression (ALP, OCN, COL1 and Runx2) of human bone marrow stromal cells. Our results suggest that the preparation of self-assembled Ca-P/polydopamine composite nanolayers is a viable method to modify the surface of biomaterials by

  8. Vulnerability and adaptation to water scarcity in the European Alps

    NASA Astrophysics Data System (ADS)

    Isoard, S.; McCallum, S.; Prutsch, A.; Benno Hain, B.; Schauser, I.

    2009-04-01

    The European Environment Agency (EEA) has recently undertaken a project addressing vulnerability and adaptation to water availability in the European Alps. Mountains are indeed one of the most vulnerable regions to climate change in Europe (EEA 2008, IPCC 2007).The Alps, in particular, can be presented as the ‘water towers' of Europe (the amount of water delivered by the Alps allocates 40% of EU consumption) where changes in water availability affect all socio-economical sectors. This therefore makes adaptation actions a regional topic with an outstanding European dimension. The specific objectives of the study were to highlight the importance of the Alps in their function as ‘water towers' for Europe and analyse the vulnerability of the Alpine Region with regard to impacts of climate change (but also to global change as a whole) focussing on water availability. Given the EU and regional policy background with regard to adaptation and water issues, the study assessed the possible needs, constraints and opportunities for adaptation to the adverse impacts for various sectors pending on water resources. Findings of this activity expanded the knowledge base, fed into the preparation of European Commission's 2009 White Paper and the Alpine Convention 2009 Report on the State of the Alps, and complemented other recent studies (e.g. OECD 2007, European Parliament Committee on Agriculture & Rural Development 2008). The method used for the study relied on the one hand on findings from recent key publications on climate change impacts (EEA 2008, IPCC 2007) and EU research projects (e.g. ClimChAlp, ProClim); on the other side it was based on selected case studies chosen within the four climatic regions of the eight Alpine countries for which an extensive series of interviews with local and regional stakeholders and decision makers has been undertaken. The interviewees had been directly involved in designing and implementing water availability-related adaptation measures

  9. The AlpArray Seismic Network: status and operation

    NASA Astrophysics Data System (ADS)

    Hetényi, György; Molinari, Irene; Clinton, John; Kissling, Edi

    2017-04-01

    The AlpArray initiative (http://www.alparray.ethz.ch) is a large-scale European collaboration to study the entire Alpine orogen at high resolution and in 3D with a large variety of geoscientific methods. The core element of the initiative is an extensive and dense broadband seismological network, the AlpArray Seismic Network (AASN). Over 300 temporary stations complement the permanent seismological stations to ensure homogeneous coverage of the greater Alpine area. The AASN has officially started operation in January 2016 and is now complete on land. It is operated in a joint effort by a number of institutions from Austria, Bosnia-Herzegovina, Croatia, Czech Republic, France, Germany, Hungary, Italy, Slovakia and Switzerland. In the Ligurian Sea, a 32-station OBS campaign is planned from June 2017 until March 2018. This will complete the coverage of the greater Alpine area at an unprecedented resolution. In this poster we present the actual status of the deployment, the effort undertaken by the contributing groups, station performance, best practices, data management as well as often encountered challenges, and provide a meeting and discussion point during the conference.

  10. Intestinal alkaline phosphatase regulates protective surface microclimate pH in rat duodenum.

    PubMed

    Mizumori, Misa; Ham, Maggie; Guth, Paul H; Engel, Eli; Kaunitz, Jonathan D; Akiba, Yasutada

    2009-07-15

    Regulation of localized extracellular pH (pH(o)) maintains normal organ function. An alkaline microclimate overlying the duodenal enterocyte brush border protects the mucosa from luminal acid. We hypothesized that intestinal alkaline phosphatase (IAP) regulates pH(o) due to pH-sensitive ATP hydrolysis as part of an ecto-purinergic pH regulatory system, comprised of cell-surface P2Y receptors and ATP-stimulated duodenal bicarbonate secretion (DBS). To test this hypothesis, we measured DBS in a perfused rat duodenal loop, examining the effect of the competitive alkaline phosphatase inhibitor glycerol phosphate (GP), the ecto-nucleoside triphosphate diphosphohydrolase inhibitor ARL67156, and exogenous nucleotides or P2 receptor agonists on DBS. Furthermore, we measured perfusate ATP concentration with a luciferin-luciferase bioassay. IAP inhibition increased DBS and luminal ATP output. Increased luminal ATP output was partially CFTR dependent, but was not due to cellular injury. Immunofluorescence localized the P2Y(1) receptor to the brush border membrane of duodenal villi. The P2Y(1) agonist 2-methylthio-ADP increased DBS, whereas the P2Y(1) antagonist MRS2179 reduced ATP- or GP-induced DBS. Acid perfusion augmented DBS and ATP release, further enhanced by the IAP inhibitor l-cysteine, and reduced by the exogenous ATPase apyrase. Furthermore, MRS2179 or the highly selective P2Y(1) antagonist MRS2500 co-perfused with acid induced epithelial injury, suggesting that IAP/ATP/P2Y signalling protects the mucosa from acid injury. Increased DBS augments IAP activity presumably by raising pH(o), increasing the rate of ATP degradation, decreasing ATP-mediated DBS, forming a negative feedback loop. The duodenal epithelial brush border IAP-P2Y-HCO(3-) surface microclimate pH regulatory system effectively protects the mucosa from acid injury.

  11. Investigations on socio economic indicators of French Alps ski industry from an explicit spatial modelling of managed snow on ski slopes

    NASA Astrophysics Data System (ADS)

    Spandre, Pierre; François, Hugues; Morin, Samuel; George-Marcelpoil, Emmanuelle; Lafaysse, Matthieu

    2017-04-01

    Investigations of the capacity of ski resorts to anticipate, cope with and recover from the impact of natural snow scarcity through snow management (grooming, snowmaking) have been realized in most of the major regions in terms of international ski offer although not in the French Alps hitherto. The present work therefore introduces an innovative approach for the investigation of socio economic implications of changes in snow conditions for the French Alps ski resorts based on a panel of 129 resorts representing 96% of the total French Alps ski lifts infrastructures. We integrated detailed spatial representations of ski resorts (including priority areas for snowmaking equipment) along with physically based snowpack modelling (including the physical impact of grooming and snowmaking). The viability of ski resorts was further adressed thanks to a commonly used rule based on the snow season duration at the village and ski lifts average elevations along with the development of original viability indicators of snow conditions in the French Alps ski resorts based on the specific periods for the economic success of winter sports: Christmas and February school holidays. Such indicators were correlated to the number of ski lifts tickets sales over the 2001 - 2014 period and proved to be relevant to investigate and predict the evolutions of ski lifts tickets sales under the current ski market conditions in the French Alps. Our results outlined the contrast of snow conditions between French Alps ski resorts, even when accounting for snow management, particularly regarding the geographical location of resorts (Southern versus Northern Alps), the size and related elevation range of ski resorts. Our physically based approach also allowed to compute the water and energy requirements for the production of Machine Made snow since the start of the development of snowguns in the French Alps. Our computations proved to be strongly correlated to the observed amounts of water from the

  12. Seismic signature of the Alpine indentation, evidence from the Eastern Alps

    PubMed Central

    Bianchi, I.; Bokelmann, G.

    2014-01-01

    The type of collision between the European and the Adriatic plates in the easternmost Alps is one of the most interesting questions regarding the Alpine evolution. Tectonic processes such as compression, escape and uplift are interconnected and shape this area. We can understand these ongoing processes better, if we look for signs of the deformation within the Earth's deep crust of the region. By collecting records from permanent and temporary seismic networks, we assemble a receiver function dataset, and analyze it with the aim of giving new insights on the structure of the lower crust and of the shallow portion of the upper mantle, which are inaccessible to direct observation. Imaging is accomplished by performing common conversion depth stacks along three profiles that crosscut the Eastern Alpine orogen, and allow isolating features consistently persistent in the area. The study shows a moderately flat Moho underlying a seismically anisotropic middle-lower crust from the Southern Alps to the Austroalpine nappes. The spatial progression of anisotropic axes reflects the orientation of the relative motion and of the stress field detected at the surface. These observations suggest that distributed deformation is due to the effect of the Alpine indentation. In the shallow upper mantle right below the Moho interface, a further anisotropic layer is recognized, extended from the Bohemian Massif to the Northern Calcareous Alps. PMID:26525181

  13. Seismic signature of the Alpine indentation, evidence from the Eastern Alps.

    PubMed

    Bianchi, I; Bokelmann, G

    2014-12-01

    The type of collision between the European and the Adriatic plates in the easternmost Alps is one of the most interesting questions regarding the Alpine evolution. Tectonic processes such as compression, escape and uplift are interconnected and shape this area. We can understand these ongoing processes better, if we look for signs of the deformation within the Earth's deep crust of the region. By collecting records from permanent and temporary seismic networks, we assemble a receiver function dataset, and analyze it with the aim of giving new insights on the structure of the lower crust and of the shallow portion of the upper mantle, which are inaccessible to direct observation. Imaging is accomplished by performing common conversion depth stacks along three profiles that crosscut the Eastern Alpine orogen, and allow isolating features consistently persistent in the area. The study shows a moderately flat Moho underlying a seismically anisotropic middle-lower crust from the Southern Alps to the Austroalpine nappes. The spatial progression of anisotropic axes reflects the orientation of the relative motion and of the stress field detected at the surface. These observations suggest that distributed deformation is due to the effect of the Alpine indentation. In the shallow upper mantle right below the Moho interface, a further anisotropic layer is recognized, extended from the Bohemian Massif to the Northern Calcareous Alps.

  14. PHENOALP: a new project on phenology in the Western Alps

    NASA Astrophysics Data System (ADS)

    Cremonese, E.

    2009-04-01

    PHENOALP is a new EU co-funded Interreg Project under the operational programme for cross-border cooperation "Italy-France (Alps-ALCOTRA)" 2007 - 2013, aiming to get a better understanding of phenological changes in the Alps. The major goals of the project are: 1- The implementation of an observation network in the involved territories (i.e. the Aosta Valley and the Savoies in the Western Alps); 2- The definition of a common observation strategy and common protocols; 3- The involvement of local community members (e.g. through schools) in the observation activities as a way to increase the awareness on the issue of the effects of climate change. Project leader is the Environmental Protection Agency of Aosta Valley (ARPA Valle d'Aosta - IT) and the partners are the Research Center on High Altitude Ecosystem (CREA - FR), Mont Avic Regional Parc (IT), Bauges Massif Regional Natural Parc (FR) and the Protected Area Service of Aosta Valley (IT). Project activities are: 1. Pheno-plantes: definition of common observation protocols (e.g. field observation and webcams) of different alpine species (trees and herbaceous) and implementation of the observation network; analysis of the relations between climate and phenological events; application and evaluation of phenological models. 2. Pheno-detection: remote sensing of European larch and high elevation pastures with MODIS data; multitemporal analysis (2000-2011) of phenological variations in the Western Alps. 3. Pheno-flux: analysis of the relation between the seasonal and interannual variability of plant phenology and productivity, assessed measuring CO2 fluxes (eddy-covariance technique), radiometric indexes and phenological events at specific (European larch stand and alpine pastures) monitoring site. 4. Pheno-zoo: definition of common observation protocols for the phenology of animal taxa (birds, mammals, amphibians and insects) along altitudinal gradients; implementation of the observation network. 5. Inter

  15. Association between serum alkaline phosphatase and primary resistance to erythropoiesis stimulating agents in chronic kidney disease: a secondary analysis of the HERO trial.

    PubMed

    Badve, Sunil V; Zhang, Lei; Coombes, Jeff S; Pascoe, Elaine M; Cass, Alan; Clarke, Philip; Ferrari, Paolo; McDonald, Stephen P; Morrish, Alicia T; Pedagogos, Eugenie; Perkovic, Vlado; Reidlinger, Donna; Scaria, Anish; Walker, Rowan; Vergara, Liza A; Hawley, Carmel M; Johnson, David W

    2015-01-01

    Erythropoiesis stimulating agent (ESA)-resistant anemia is common in chronic kidney disease (CKD). To evaluate the determinants of severity of ESA resistance in patients with CKD and primary ESA-resistance. Secondary analysis of a randomized controlled trial (the Handling Erythropoietin Resistance with Oxpentifylline, HERO). 53 adult patients with CKD stage 4 or 5 and primary ESA-resistant anemia (hemoglobin ≤120 g/L, ESA resistance index [ERI] ≥1.0 IU/kg/week/gHb for erythropoietin or ≥0.005 μg/kg/week/gHb for darbepoeitin, no cause for ESA-resistance identified). Iron studies, parathyroid hormone, albumin, liver enzymes, phosphate or markers of oxidative stress and inflammation. Participants were divided into tertiles of ERI. Multinomial logistic regression was used to analyse the determinants of ERI tertiles. All patients, except one, were receiving dialysis for end-stage kidney disease. The mean ± SD ERI values in the low (n = 18), medium (n = 18) and high (n = 17) ERI tertiles were 1.4 ± 0.3, 2.3 ± 0.2 and 3.5 ± 0.8 IU/kg/week/gHb, respectively (P < 0.001). There were no significant differences observed in age, gender, ethnicity, cause of kidney disease, diabetes, iron studies, parathyroid hormone, albumin, liver enzymes, phosphate or markers of oxidative stress and inflammation between the ERI tertiles. The median [inter-quartile range] serum alkaline phosphatase concentrations in the low, medium and high ERI tertiles were 89 [64,121], 99 [76,134 and 148 [87,175] U/L, respectively (P = 0.054). There was a weak but statistically significant association between ERI and serum alkaline phosphatase (R(2) = 0.06, P = 0.03). Using multinomial logistic regression, the risk of being in the high ERI tertile relative to the low ERI tertile increased with increasing serum alkaline phosphatase levels (P = 0.02). No other variables were significantly associated with ERI. Small sample size; bone

  16. Adapting the ALP Model for Student and Institutional Needs

    ERIC Educational Resources Information Center

    Sides, Meredith

    2016-01-01

    With the increasing adoption of accelerated models of learning comes the necessary step of adapting these models to fit the unique needs of the student population at each individual institution. One such college adapted the ALP (Accelerated Learning Program) model and made specific changes to the target population, structure and scheduling, and…

  17. [A novel tissue-engineered bone constructed by using human adipose-derived stem cells and biomimetic calcium phosphate scaffold coprecipitated with bone morphogenetic protein-2].

    PubMed

    Jiang, W R; Zhang, X; Liu, Y S; Wu, G; Ge, Y J; Zhou, Y S

    2017-02-18

    To construct a novel biomimetic calcium phosphate (BioCaP) scaffold loaded with bone morphogenetic protein-2 (BMP-2), and to investigate its role in the osteogenesis of human adipose-derived stem cells (hASCs) in vitro and in vivo. The BioCaP scaffold coprecipitated with BMP-2 (BMP-2-BioCaP) was constructed in this study. Field emission scanning electron microscopy (SEM) was used to analyze the morphology of the surfaces. The release kinetics was measured to evaluate the slow-release characteristics in vitro. BMP-2-BioCaP was immersed in proliferation medium (PM) or osteogenic medium (OM), respectively. The supernatants were collected and used to culture hASCs in vitro. Cell numbers were determined using the cell-counting kit-8 (CCK-8) to assess the cell proliferation. After 7 and 14 days, alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP. After 14 and 21 days, the calcification deposition was determined by alizarin red S (ARS) staining and quantification. The expressions of the osteoblast-related genes were tested on day 4 and day 14. In the in vivo study, 6 nude mice were used and implanted subcutaneously into the back of the nude mice for 4 groups: (1) BioCaP scaffold only, (2) BioCaP scaffold+hASCs, (3) BMP-2-BioCaP scaffold, (4) BMP-2-BioCaP scaffold+hASCs (test group). After 4 weeks of implantation, hematoxylin-eosin (HE) staining was performed to evaluate the in vivo osteogenesis of hASCs. SEM observations showed that BioCaP and BMP-2-BioCaP scaffold were entirely composed of straight, plate-like and sharp-edged crystal units, and the length of the crystal units varied between 5 and 10 μm. Release kinetics analysis demonstrated that BMP-2 incorporated with BioCaP could be released at certain concentration and last for more than 21 days, and the accumulative protein release could reach 20%. CCK-8 assays showed that cell proliferation was not significantly affected by BMP-2-BioCaP. ALP activity was higher by the

  18. Collision in the Central Alps: 1. Thermal Modelling

    NASA Astrophysics Data System (ADS)

    Engi, M.; Roselle, G. T.; Brouwer, F. M.; Berger, A.

    2003-04-01

    Recent tectonic reconstructions for the Central Alps, based in part on seismic profiles across of the orogen, have produced fairly robust kinematic scenarios for the Tertiary evolution. We have used these to set up 2D finite element models [1] to simulate the thermal evolution at orogenic scales. Results are helpful to understand the metamorphic and geochronological record in the Central Alps. Several features recognized as crucial in collisional orogens have been incorporated in our models: Adaptive grids are used to accommodate tectonic mass flow; properties of a tectonic accretion channel (TAC), situated near the footwall of the upper (Apulian) plate, are incorporated (TAC: 5-10 km wide [2]); a mobile fragment (pit: 5-10 km thick, 25-50 km long) at the plate interface is allowed to first be subducted, then to be extruded along the subduction channel to mid-crustal levels during the nappe stacking phase, and finally to be exhumed by backthrusting and erosion; partial melting and its thermal effects are computed. The thermal evolution in crucial parts of the model orogen is depicted in P-T and T-t trajectories, and in time slices showing the evolution of metamorphic facies and degrees of late partial melting. Comparison of simulation results with the regional distribution of (Eocene) high pressure fragments in the Lepontine Alps and of their (Oligo/Miocene) Barrovian overprint indicate that (a) decompression is near-isothermal along a very imited part of the path only; (b) the highest temperatures attained following collision do not reach the observed ˜700^oC unless the TAC is fairly radiogenic (heat production ge˜2 μW/m^3) or there is substantial heat advected by asthenospheric melts migrating up the subduction channel; (c) moderate amounts of partial melting occurred within the the TAC during decompression, following the assembly (at mid-crustal levels) of various crustal and mantle fragments with very diverse P-T-t histories. [1] Roselle et al. (2002) Amer

  19. Medical ethnobotany of the Albanian Alps in Kosovo.

    PubMed

    Mustafa, Behxhet; Hajdari, Avni; Krasniqi, Feriz; Hoxha, Esat; Ademi, Hatixhe; Quave, Cassandra L; Pieroni, Andrea

    2012-01-28

    Ethnobotanical studies are crucial in South-Eastern Europe for fostering local development and also for investigating the dynamics of Traditional Environmental Knowledge (TEK) related to plants in one of the most crucial European hotspots for biocultural diversity. The current medico-ethnobotanical survey was conducted in rural alpine communities in Kosovo. The aims of the study were twofold: 1) to document the state of TEK of medicinal plants in these communities; 2) to compare these findings with that of similar field studies previously conducted among local populations inhabiting the Montenegrin and Albanian side of the same Alpine range. Field research was conducted in 36 villages on the Kosovar side of the Albanian Alps. Snowball sampling techniques were used to recruit 91 elderly informants (≥ 50 years-old) for participation in semi-structured interviews and structured surveys regarding the use of the local flora for medicinal and food purposes. Standard ethnobotanical methods were employed and prior informed consent was obtained for all study participants. The uses of 98 plants species belonging to 39 families were recorded; the most quoted botanical families were Rosaceae, Asteraceae, and Lamiaceae. Mainly decoctions and infusions were quoted as folk medicinal preparations and the most common uses referred to gastrointestinal and respiratory disorders, as well as illnesses of the uro-genital system. Among the most uncommon medicinal taxa quoted by the informants, Carduus nutans L., Echinops bannaticus Rochel ex Schrad., and Orlaya grandiflora Hoffm. may merit phytochemical and phytopharmacological investigations.Comparison of the data with other ethnobotanical field studies recently conducted on the Albanian and Montenegrin sides of the same Alps has shown a remarkable link between the medical ethnobotany of Montenegrin and Kosovar side of the Albanian Alps. Moreover, folk uses of the most quoted wild medicinal taxa recorded in Kosovo often include those

  20. Microbial solubilization of phosphate

    DOEpatents

    Rogers, R.D.; Wolfram, J.H.

    1993-10-26

    A process is provided for solubilizing phosphate from phosphate containing ore by treatment with microorganisms which comprises forming an aqueous mixture of phosphate ore, microorganisms operable for solubilizing phosphate from the phosphate ore and maintaining the aqueous mixture for a period of time and under conditions operable to effect the microbial solubilization process. An aqueous solution containing soluble phosphorus can be separated from the reacted mixture by precipitation, solvent extraction, selective membrane, exchange resin or gravity methods to recover phosphate from the aqueous solution. 6 figures.

  1. Microbial solubilization of phosphate

    DOEpatents

    Rogers, Robert D.; Wolfram, James H.

    1993-01-01

    A process is provided for solubilizing phosphate from phosphate containing ore by treatment with microorganisms which comprises forming an aqueous mixture of phosphate ore, microorganisms operable for solubilizing phosphate from the phosphate ore and maintaining the aqueous mixture for a period of time and under conditions operable to effect the microbial solubilization process. An aqueous solution containing soluble phosphorous can be separated from the reacted mixture by precipitation, solvent extraction, selective membrane, exchange resin or gravity methods to recover phosphate from the aqueous solution.

  2. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 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 (granular...

  3. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 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 (granular...

  4. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 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 (granular...

  5. Use of Fe3O4 Nanoparticles for Enhancement of Biosensor Response to the Herbicide 2,4-Dichlorophenoxyacetic Acid

    PubMed Central

    Loh, Kee-Shyuan; Lee, Yook Heng; Musa, Ahmad; Salmah, Abdul Aziz; Zamri, Ishak

    2008-01-01

    Magnetic nanoparticles of Fe3O4 were synthesized and characterized using transmission electron microscopy and X-ray diffraction. The Fe3O4 nanoparticles were found to have an average diameter of 5.48 ±1.37 nm. An electrochemical biosensor based on immobilized alkaline phosphatase (ALP) and Fe3O4 nanoparticles was studied. The amperometric biosensor was based on the reaction of ALP with the substrate ascorbic acid 2-phosphate (AA2P). The incorporation of the Fe3O4 nanoparticles together with ALP into a sol gel/chitosan biosensor membrane has led to the enhancement of the biosensor response, with an improved linear response range to the substrate AA2P (5-120 μM) and increased sensitivity. Using the inhibition property of the ALP, the biosensor was applied to the determination of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). The use of Fe3O4 nanoparticles gives a two-fold improvement in the sensitivity towards 2,4-D, with a linear response range of 0.5-30 μgL-1. Exposure of the biosensor to other toxicants such as heavy metals demonstrated only slight interference from metals such as Hg2+, Cu2+, Ag2+ and Pb2+. The biosensor was shown to be useful for the determination of the herbicide 2, 4-D because good recovery of 95-100 percent was obtained, even though the analysis was performed in water samples with a complex matrix. Furthermore, the results from the analysis of 2,4-D in water samples using the biosensor correlated well with a HPLC method. PMID:27873839

  6. The lichens of the Alps – an annotated checklist

    PubMed Central

    Nimis, Pier Luigi; Hafellner, Josef; Roux, Claude; Clerc, Philippe; Helmut Mayrhofer; Martellos, Stefano; Bilovitz, Peter O.

    2018-01-01

    Abstract This is the first attempt to provide an overview of the lichen diversity of the Alps, one of the biogegraphically most important and emblematic mountain systems worldwide. The checklist includes all lichenised species, plus a set of non- or doubtfully lichenised taxa frequently treated by lichenologists, excluding non-lichenised lichenicolous fungi. Largely based on recent national or regional checklists, it provides a list of all infrageneric taxa (with synonyms) hitherto reported from the Alps, with data on their distribution in eight countries (Austria, France, Germany, Liechtenstein, Monaco, Italy, Slovenia, Switzerland) and in 42 Operational Geographic Units, mostly corresponding to administrative subdivisions within the countries. Data on the main substrates and on the altitudinal distribution are also provided. A short note points to the main ecological requirements of each taxon and/or to open taxonomic problems. Particularly poorly known taxa are flagged and often provided with a short description, to attract the attention of specialists. The total number of infrageneric taxa is 3,163, including 117 non- or doubtfully lichenised taxa. The richness of the lichen biota fairly well corresponds with the percent of the Alpine area occupied by each country: Austria (2,337 taxa), Italy (2,169), France (2,028), Switzerland (1,835), Germany (1,168), Slovenia (890) and Lichtenstein (152), no lichen having ever been reported from Monaco. The number of poorly known taxa is quite high (604, 19.1% of the total), which indicates that, in spite of the Alps being one of the lichenologically most studied mountain systems worldwide, much work is still needed to reach a satisfactory picture of their real lichen diversity. Thirteen new combinations are proposed in the genera Agonimia, Aspicilia, Bagliettoa, Bellemerea, Carbonea, Lepra, Miriquidica, Polysporina, Protothelenella, Pseudosagedia and Thelidium. PMID:29706791

  7. [Reconstruction of maxillary sinus superior wall fractures with calcium phosphate cement/recombinant human bonemorphogenetic protein 7 compound implanted material in rabbit].

    PubMed

    Zhang, Qunhui; Yu, Feng; Zhang, Haoliang; Gong, Huicheng; Lin, Ying

    2015-11-01

    To evaluate the osteogenetic character and repairing maxillary sinus superior wall fractures capability of calcium phosphate cement (CPC) before and after combined with recombinant human bone morphogenetie protein-7(rhBMP-7). A 10 mmX5 mm bone defect in the maxillary sinus superior wall was induced by surgery in all 24 New Zealand white rabbits. These 24 rabbits were randomly divided into two groups. The defects were repaired with CPC group (n = 12) and CPC/rhBMP-7 group (n = 12). The osteogenesis of bone defect was monitored by gro'ss observation, histological examination, observation under scanning electron microscope and measurement of ALP activity at 6 and 12 weeks after the implantation. In group CPC,new bone was found to form slowly and little by little. In group CPC/rhBMP-7, however, new bone was observed to form early and massively. The ALP activity in group CPC showed significant statistical difference with that of group CPC/rhBMP-7 (P < 0.05). The CPC/rhBMP-7 composite has osteoconductibility and osteoinductibility, comparing the use of CPC/rhBMP-7 with CPC for the repair of orbital fracture, the former show obvious advantage repairing ability in maxillary sinus superior wall defect.

  8. Application of perfusion culture system improves in vitro and in vivo osteogenesis of bone marrow-derived osteoblastic cells in porous ceramic materials.

    PubMed

    Wang, Yichao; Uemura, Toshimasa; Dong, Jian; Kojima, Hiroko; Tanaka, Junzo; Tateishi, Tetsuya

    2003-12-01

    Composites of bone marrow-derived osteoblasts (BMOs) and porous ceramics have been widely used as a bone graft model for bone tissue engineering. Perfusion culture has potential utility for many cell types in three-dimensional (3D) culture. Our hypothesis was that perfusion of medium would increase the cell viability and biosynthetic activity of BMOs in porous ceramic materials, which would be revealed by increased levels of alkaline phosphate (ALP) activity and osteocalcin (OCN) and enhanced bone formation in vivo. For testing in vitro, BMO/beta-tricalcium phosphate composites were cultured in a perfusion container (Minucells and Minutissue, Bad Abbach, Germany) with fresh medium delivered at a rate of 2 mL/h by a peristaltic pump. The ALP activity and OCN content of composites were measured at the end of 1, 2, 3, and 4 weeks of subculture. For testing in vivo, after subculturing for 2 weeks, the composites were subcutaneously implanted into syngeneic rats. These implants were harvested 4 or 8 weeks later. The samples then underwent a biochemical analysis of ALP activity and OCN content and were observed by light microscopy. The levels of ALP activity and OCN in the composites were significantly higher in the perfusion group than in the control group (p < 0.01), both in vitro and in vivo. Histomorphometric analysis of the hematoxylin- and eosin-stained sections revealed a higher average ratio of bone to pore in BMO/beta-TCP composites of the perfusion group after implantation: 47.64 +/- 6.16 for the perfusion group and 26.22 +/- 4.84 for control at 4 weeks (n = 6, p < 0.01); 67.97 +/- 3.58 for the perfusion group and 47.39 +/- 4.10 for control at 8 weeks (n = 6, p < 0.05). These results show that the application of a perfusion culture system during the subculture of BMOs in a porous ceramic scaffold is beneficial to their osteogenesis. After differentiation culture in vitro with the perfusion culture system, the activity of the osteoblastic cells and the

  9. French Alps, Mont Blanc, French/Italian Border

    NASA Image and Video Library

    1992-04-02

    In this southeast looking view, Mont Blanc, on the French/Italian border, (48.0N, 4.5E) the highest mountain peak in all of Europe, is just below and right of center (below the end of the prominent valley of the Aosta River, in the center of the photo. The rivers flow out of the Alps into Italy toward Turin. Chamonix, the famous resort town and center of Alpine mountain climbing, lies in the valley just below Mont Blanc.

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

    NASA Astrophysics Data System (ADS)

    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.

  11. Nucleotide sequences encoding a thermostable alkaline protease

    DOEpatents

    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.

  12. Experimental and first-principles calculation study of the pressure-induced transitions to a metastable phase in GaP O4 and in the solid solution AlP O4-GaP O4

    NASA Astrophysics Data System (ADS)

    Angot, E.; Huang, B.; Levelut, C.; Le Parc, R.; Hermet, P.; Pereira, A. S.; Aquilanti, G.; Frapper, G.; Cambon, O.; Haines, J.

    2017-08-01

    α -Quartz-type gallium phosphate and representative compositions in the AlP O4-GaP O4 solid solution were studied by x-ray powder diffraction and absorption spectroscopy, Raman scattering, and by first-principles calculations up to pressures of close to 30 GPa. A phase transition to a metastable orthorhombic high-pressure phase along with some of the stable orthorhombic C m c m CrV O4 -type material is found to occur beginning at 9 GPa at 320 ∘C in GaP O4 . In the case of the AlP O4-GaP O4 solid solution at room temperature, only the metastable orthorhombic phase was obtained above 10 GPa. The possible crystal structures of the high-pressure forms of GaP O4 were predicted from first-principles calculations and the evolutionary algorithm USPEX. A predicted orthorhombic structure with a P m n 21 space group with the gallium in sixfold and phosphorus in fourfold coordination was found to be in the best agreement with the combined experimental data from x-ray diffraction and absorption and Raman spectroscopy. This method is found to very powerful to better understand competition between different phase transition pathways at high pressure.

  13. SKS splitting results in central Italy and Dinaric region inside the AlpArray-CASE project

    NASA Astrophysics Data System (ADS)

    Salimbeni, S.; Prevolnik, S.; Pondrelli, S.; Molinari, I.; Stipcevic, J.; Kissling, E.; Šipka, V.; Herak, M.

    2017-12-01

    In the framework of the AlpArray project (AlpArray Seismic Network, 2015), the complementary "Central Adriatic Seismic Experiment" (CASE; AlpArray Seismic Network, 2016) was established as collaboration between ETH Zürich, University of Zagreb, INGV and Republic Hydrometeorological Service of Republic of Srpska. The CASE project consists of 9 temporary stations, installed in October 2016, located in Bosnia and Herzegovina, Croatia and Italy. Temporary broadband seismic stations, with the permanent stations present in the region shared by the Croatian Seismological Service and INGV, make an almost continuous transect cutting the Central-Southern Appenines, the central Adriatic region, central External Dinarides and finishing at the eastern margin of the Internal Dinarides. The presence of the the Apenninic and Dinarides slabs, verging in opposite directions and plunging along the opposite sides of the Adriatic plate, make this area a peculiar spot to understand the complex dynamic of the region. Various tomographic images (e.g. Bijwaard and Spakman, 2000; Piromallo and Morelli, 2003) shows not continuous slabs under the Appenines and the Dinarides, suggesting the presence of slab-gaps right beneath the region covered by the CASE experiment. Here we present the preliminary results of the SKS splitting analysis performed on the data recorded by the temporary and permanent seismic stations included in the CASE project. The new results, in combination with previous interpretation, will provide clues about how Northern and Southern Apennines are connected at depth, how the slab rollback of the Apennines thrust belt acted and if and how the Apennines are in relation with the Dinaric region. Together with the measurements from previous studies and from the AlpArray project, our new data will support the mapping of the seismic anisotropy deformation pattern from Western Alps to Pannonian region.

  14. Availability of phosphate for phytoplankton and bacteria and of labile organic carbon for bacteria at different pCO2 levels in a mesocosm study

    NASA Astrophysics Data System (ADS)

    Tanaka, T.; Thingstad, T. F.; Løvdal, T.; Grossart, H.-P.; Larsen, A.; Schulz, K. G.; Riebesell, U.

    2007-11-01

    Availability of phosphate for phytoplankton and bacteria and of labile organic carbon for bacteria at different pCO2 levels were studied in a mesocosm experiment (PeECE III). Using nutrient-depleted SW Norwegian fjord waters, three different levels of pCO2 (350 μatm: 1×CO2; 750 μatm: 2×CO2; 1050 μatm: 3×CO2) were set up, and nitrate and phosphate were added at the start of the experiment in order to induce a phytoplankton bloom. Despite similar responses of total particulate P concentration and phosphate turnover time at the three different pCO2 levels, the size distribution of particulate P and 33PO4 uptake suggested that phosphate transferred to the >10 μm fraction was greater in the 3×CO2 mesocosm during the first 6-10 days when phosphate concentration was high. During the period of phosphate depletion (after Day 12), specific phosphate affinity and specific alkaline phosphatase activity (APA) suggested a P-deficiency (i.e. suboptimal phosphate supply) but not a P-limitation for the phytoplankton and bacterial community at the three different pCO2 levels. Although specific phosphate affinity and specific APA tended to be higher in 3×CO2 than in 2×CO2 and 1×CO2 mesocosms during the phosphate depletion period, no statistical differences were found. Responses of specific glucose affinity for bacteria were similar at the three different pCO2 levels. Measured specific glucose affinities were consistently much lower than the theoretical maximum predicted from the diffusion-limited model, suggesting that bacterial growth was not limited by the availability of labile dissolved organic carbon. These results suggest that availability of phosphate and glucose was similar at the three different pCO2 levels.

  15. Nucleotide sequences encoding a thermostable alkaline protease

    DOEpatents

    Wilson, D.B.; Lao, G.

    1998-01-06

    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. 3 figs.

  16. Analog Modeling of the Interplay between Subduction and Lateral Extrusion in the European Alps

    NASA Astrophysics Data System (ADS)

    van Gelder, I. E.; Willingshofer, E.; Sokoutis, D.

    2014-12-01

    In the European Alps lateral extrusion is traditionally viewed as a lithospheric scale process that is related to northward indentation of a weak orogenic wedge (the eastern Alps) by a rigid indenter in upper plate position (the Adriatic plate). Critical for the efficiency of the extrusion process is the presence of a 'free boundary' at high angle to the indentation direction. The 'free boundary' in the eastern Alps is the result of the eastward extending Pannonian realm synchronous to indentation. However, indentation has become debatable as recent high-resolution tomography suggests that the Adriatic mantle lithosphere subducted under the extruding Alps. These findings raise first order questions related to: (a) the partitioning of deformation between lateral extrusion of the upper plate and coeval subduction of Adria, (b) the rheology of the lower and upper plates, and (c) the rheology of the plate contact controlling the amount of extrusion on the upper plate vs. accretion on the lower plate.In this analog modeling study, we couple for the first time lateral extrusion tectonics to subduction of the lower plate; thus, extrusion taking place in the upper plate. Within the lithospheric scale models, the lithospheres of the two plates are weakly coupled along an inclined boundary and have contrasting mantle lithosphere strength (stronger in the subducting plate). The interplay of extrusion vs subduction is inferred by varying the mechanical boundary conditions, e.g. the degree of resistance at the 'unconstrained' margin, the strength contrast between the upper and the lower plates and the width of the indented region.The experimental results emphasize that extrusion in the eastern Alps is compatible with coeval subduction of the Adriatic plate. The first experimental series suggests that the following mechanical conditions play a key role in the interplay between extrusion and subduction: (a) the extruding plate is weaker than the subducting plate, (b) the plate

  17. On-demand Simulation of Atmospheric Transport Processes on the AlpEnDAC Cloud

    NASA Astrophysics Data System (ADS)

    Hachinger, S.; Harsch, C.; Meyer-Arnek, J.; Frank, A.; Heller, H.; Giemsa, E.

    2016-12-01

    The "Alpine Environmental Data Analysis Centre" (AlpEnDAC) develops a data-analysis platform for high-altitude research facilities within the "Virtual Alpine Observatory" project (VAO). This platform, with its web portal, will support use cases going much beyond data management: On user request, the data are augmented with "on-demand" simulation results, such as air-parcel trajectories for tracing down the source of pollutants when they appear in high concentration. The respective back-end mechanism uses the Compute Cloud of the Leibniz Supercomputing Centre (LRZ) to transparently calculate results requested by the user, as far as they have not yet been stored in AlpEnDAC. The queuing-system operation model common in supercomputing is replaced by a model in which Virtual Machines (VMs) on the cloud are automatically created/destroyed, providing the necessary computing power immediately on demand. From a security point of view, this allows to perform simulations in a sandbox defined by the VM configuration, without direct access to a computing cluster. Within few minutes, the user receives conveniently visualized results. The AlpEnDAC infrastructure is distributed among two participating institutes [front-end at German Aerospace Centre (DLR), simulation back-end at LRZ], requiring an efficient mechanism for synchronization of measured and augmented data. We discuss our iRODS-based solution for these data-management tasks as well as the general AlpEnDAC framework. Our cloud-based offerings aim at making scientific computing for our users much more convenient and flexible than it has been, and to allow scientists without a broad background in scientific computing to benefit from complex numerical simulations.

  18. Mountaineering accidents in the European Alps: have the numbers increased in recent years?

    PubMed

    Lischke, V; Byhahn, C; Westphal, K; Kessler, P

    2001-01-01

    Media reports convey the impression that the incidence of fatal accidents in the European Alps has increased. Because more specific data are lacking, we analyzed available data from the mountain rescue services in Germany, Austria, southern Tirol, Zermatt/Switzerland, and Chamonix/France from 1987 until 1997. Information was gathered from the annual reports of the Austrian Mountain Rescue Service, the Swiss Alpine Club Rescue Station in Zermatt, the Mountain Rescue Service of the Southern Tirol Alpine Club, the Mountain Rescue Service of the Bavarian Red Cross, and the Department of Mountain Medicine and Traumatology from the Hospital in Chamonix. Although the total number of rescue missions and injured alpinists increased significantly during the period, the number of fatalities retrieved during such rescue missions showed no significant increase. Even taking into account the varying definitions of "mountain accident" used in these countries, available data from the analyzed areas of the European Alps do not demonstrate a drastic increase in the number of fatalities. In the future, data concerning mountain accidents in the European Alps should be monitored according to standard definitions and stored by the International Commission for Alpine Rescue.

  19. The AlpArray Seismic Network: A Large-Scale European Experiment to Image the Alpine Orogen

    NASA Astrophysics Data System (ADS)

    Hetényi, György; Molinari, Irene; Clinton, John; Bokelmann, Götz; Bondár, István; Crawford, Wayne C.; Dessa, Jean-Xavier; Doubre, Cécile; Friederich, Wolfgang; Fuchs, Florian; Giardini, Domenico; Gráczer, Zoltán; Handy, Mark R.; Herak, Marijan; Jia, Yan; Kissling, Edi; Kopp, Heidrun; Korn, Michael; Margheriti, Lucia; Meier, Thomas; Mucciarelli, Marco; Paul, Anne; Pesaresi, Damiano; Piromallo, Claudia; Plenefisch, Thomas; Plomerová, Jaroslava; Ritter, Joachim; Rümpker, Georg; Šipka, Vesna; Spallarossa, Daniele; Thomas, Christine; Tilmann, Frederik; Wassermann, Joachim; Weber, Michael; Wéber, Zoltán; Wesztergom, Viktor; Živčić, Mladen

    2018-04-01

    The AlpArray programme is a multinational, European consortium to advance our understanding of orogenesis and its relationship to mantle dynamics, plate reorganizations, surface processes and seismic hazard in the Alps-Apennines-Carpathians-Dinarides orogenic system. The AlpArray Seismic Network has been deployed with contributions from 36 institutions from 11 countries to map physical properties of the lithosphere and asthenosphere in 3D and thus to obtain new, high-resolution geophysical images of structures from the surface down to the base of the mantle transition zone. With over 600 broadband stations operated for 2 years, this seismic experiment is one of the largest simultaneously operated seismological networks in the academic domain, employing hexagonal coverage with station spacing at less than 52 km. This dense and regularly spaced experiment is made possible by the coordinated coeval deployment of temporary stations from numerous national pools, including ocean-bottom seismometers, which were funded by different national agencies. They combine with permanent networks, which also required the cooperation of many different operators. Together these stations ultimately fill coverage gaps. Following a short overview of previous large-scale seismological experiments in the Alpine region, we here present the goals, construction, deployment, characteristics and data management of the AlpArray Seismic Network, which will provide data that is expected to be unprecedented in quality to image the complex Alpine mountains at depth.

  20. A fluorescent graphitic carbon nitride nanosheet biosensor for highly sensitive, label-free detection of alkaline phosphatase.

    PubMed

    Xiang, Mei-Hao; Liu, Jin-Wen; Li, Na; Tang, Hao; Yu, Ru-Qin; Jiang, Jian-Hui

    2016-02-28

    Graphitic C3N4 (g-C3N4) nanosheets provide an attractive option for bioprobes and bioimaging applications. Utilizing highly fluorescent and water-dispersible ultrathin g-C3N4 nanosheets, a highly sensitive, selective and label-free biosensor has been developed for ALP detection for the first time. The developed approach utilizes a natural substrate of ALP in biological systems and thus affords very high catalytic efficiency. This novel biosensor is demonstrated to enable quantitative analysis of ALP in a wide range from 0.1 to 1000 U L(-1) with a low detection limit of 0.08 U L(-1), which is among the most sensitive assays for ALP. It is expected that the developed method may provide a low-cost, convenient, rapid and highly sensitive platform for ALP-based clinical diagnostics and biomedical applications.

  1. The alkaline solution to the emergence of life: energy, entropy and early evolution.

    PubMed

    Russell, Michael J

    2007-01-01

    The Earth agglomerates and heats. Convection cells within the planetary interior expedite the cooling process. Volcanoes evolve steam, carbon dioxide, sulfur dioxide and pyrophosphate. An acidulous Hadean ocean condenses from the carbon dioxide atmosphere. Dusts and stratospheric sulfurous smogs absorb a proportion of the Sun's rays. The cooled ocean leaks into the stressed crust and also convects. High temperature acid springs, coupled to magmatic plumes and spreading centers, emit iron, manganese, zinc, cobalt and nickel ions to the ocean. Away from the spreading centers cooler alkaline spring waters emanate from the ocean floor. These bear hydrogen, formate, ammonia, hydrosulfide and minor methane thiol. The thermal potential begins to be dissipated but the chemical potential is dammed. The exhaling alkaline solutions are frustrated in their further attempt to mix thoroughly with their oceanic source by the spontaneous precipitation of biomorphic barriers of colloidal iron compounds and other minerals. It is here we surmise that organic molecules are synthesized, filtered, concentrated and adsorbed, while acetate and methane--separate products of the precursor to the reductive acetyl-coenzyme-A pathway-are exhaled as waste. Reactions in mineral compartments produce acetate, amino acids, and the components of nucleosides. Short peptides, condensed from the simple amino acids, sequester 'ready-made' iron sulfide clusters to form protoferredoxins, and also bind phosphates. Nucleotides are assembled from amino acids, simple phosphates carbon dioxide and ribose phosphate upon nanocrystalline mineral surfaces. The side chains of particular amino acids register to fitting nucleotide triplet clefts. Keyed in, the amino acids are polymerized, through acid-base catalysis, to alpha chains. Peptides, the tenuous outer-most filaments of the nanocrysts, continually peel away from bound RNA. The polymers are concentrated at cooler regions of the mineral compartments through

  2. Catalytically active alkaline molten globular enzyme: Effect of pH and temperature on the structural integrity of 5-aminolevulinate synthase*

    PubMed Central

    Stojanovski, Bosko M.; Breydo, Leonid; Hunter, Gregory A.; Uversky, Vladimir N.; Ferreira, Gloria C.

    2014-01-01

    5-Aminolevulinate synthase (ALAS), a pyridoxal-5′phosphate (PLP)-dependent enzyme, catalyzes the first step of heme biosynthesis in mammals. Circular dichroism (CD) and fluorescence spectroscopies were used to examine the effects of pH (1.0–3.0 and 7.5–10.5) and temperature (20 and 37 °C) on the structural integrity of ALAS. The secondary structure, as deduced from far-UV CD, is mostly resilient to pH and temperature changes. Partial unfolding was observed at pH 2.0, but further decreasing pH resulted in acid-induced refolding of the secondary structure to nearly native levels. The tertiary structure rigidity, monitored by near-UV CD, is lost under acidic and specific alkaline conditions (pH 10.5 and pH 9.5/37 °C), where ALAS populates a molten globule state. As the enzyme becomes less structured with increased alkalinity, the chiral environment of the internal aldimine is also modified, with a shift from a 420 nm to 330 nm dichroic band. Under acidic conditions, the PLP cofactor dissociates from ALAS. Reaction with 8-anilino-1-naphtalenesulfonic acid corroborates increased exposure of hydrophobic clusters in the alkaline and acidic molten globules, although the reaction is more pronounced with the latter. Furthermore, quenching the intrinsic fluorescence of ALAS with acrylamide at pH 1.0 and 9.5 yielded subtly different dynamic quenching constants. The alkaline molten globule state of ALAS is catalytically active (pH 9.5/37 °C), although the kcat value is significantly decreased. Finally, the binding of 5-aminolevulinate restricts conformational fluctuations in the alkaline molten globule. Overall, our findings prove how the structural plasticity of ALAS contributes to reaching a functional enzyme. PMID:25240868

  3. Catalytically active alkaline molten globular enzyme: Effect of pH and temperature on the structural integrity of 5-aminolevulinate synthase.

    PubMed

    Stojanovski, Bosko M; Breydo, Leonid; Hunter, Gregory A; Uversky, Vladimir N; Ferreira, Gloria C

    2014-12-01

    5-Aminolevulinate synthase (ALAS), a pyridoxal-5'phosphate (PLP)-dependent enzyme, catalyzes the first step of heme biosynthesis in mammals. Circular dichroism (CD) and fluorescence spectroscopies were used to examine the effects of pH (1.0-3.0 and 7.5-10.5) and temperature (20 and 37°C) on the structural integrity of ALAS. The secondary structure, as deduced from far-UV CD, is mostly resilient to pH and temperature changes. Partial unfolding was observed at pH2.0, but further decreasing pH resulted in acid-induced refolding of the secondary structure to nearly native levels. The tertiary structure rigidity, monitored by near-UV CD, is lost under acidic and specific alkaline conditions (pH10.5 and pH9.5/37°C), where ALAS populates a molten globule state. As the enzyme becomes less structured with increased alkalinity, the chiral environment of the internal aldimine is also modified, with a shift from a 420nm to 330nm dichroic band. Under acidic conditions, the PLP cofactor dissociates from ALAS. Reaction with 8-anilino-1-naphthalenesulfonic acid corroborates increased exposure of hydrophobic clusters in the alkaline and acidic molten globules, although the reaction is more pronounced with the latter. Furthermore, quenching the intrinsic fluorescence of ALAS with acrylamide at pH1.0 and 9.5 yielded subtly different dynamic quenching constants. The alkaline molten globule state of ALAS is catalytically active (pH9.5/37°C), although the kcat value is significantly decreased. Finally, the binding of 5-aminolevulinate restricts conformational fluctuations in the alkaline molten globule. Overall, our findings prove how the structural plasticity of ALAS contributes to reaching a functional enzyme. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Effects of pH on the production of phosphate and pyrophosphate by matrix vesicles' biomimetics.

    PubMed

    Simão, Ana Maria S; Bolean, Maytê; Hoylaerts, Marc F; Millán, José Luis; Ciancaglini, Pietro

    2013-09-01

    During endochondral bone formation, chondrocytes and osteoblasts synthesize and mineralize the extracellular matrix through a process that initiates within matrix vesicles (MVs) and ends with bone mineral propagation onto the collagenous scaffold. pH gradients have been identified in the growth plate of long bones, but how pH changes affect the initiation of skeletal mineralization is not known. Tissue-nonspecific alkaline phosphatase (TNAP) degrades extracellular inorganic pyrophosphate (PPi), a mineralization inhibitor produced by ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1), while contributing Pi from ATP to initiate mineralization. TNAP and NPP1, alone or combined, were reconstituted in dipalmitoylphosphatidylcholine liposomes to mimic the microenvironment of MVs. The hydrolysis of ATP, ADP, AMP, and PPi was studied at pH 8 and 9 and compared to the data determined at pH 7.4. While catalytic efficiencies in general were higher at alkaline pH, PPi hydrolysis was maximal at pH 8 and indicated a preferential utilization of PPi over ATP at pH 8 versus 9. In addition, all proteoliposomes induced mineral formation when incubated in a synthetic cartilage lymph containing 1 mM ATP as substrate and amorphous calcium phosphate or calcium-phosphate-phosphatidylserine complexes as nucleators. Propagation of mineralization was significantly more efficient at pH 7.5 and 8 than at pH 9. Since a slight pH elevation from 7.4 to 8 promotes considerably more hydrolysis of ATP, ADP, and AMP primarily by TNAP, this small pH change facilitates mineralization, especially via upregulated PPi hydrolysis by both NPP1 and TNAP, further elevating the Pi/PPi ratio, thus enhancing bone mineralization.

  5. Metabolic changes associated with tumor metastasis, part 1: tumor pH, glycolysis and the pentose phosphate pathway.

    PubMed

    Payen, Valéry L; Porporato, Paolo E; Baselet, Bjorn; Sonveaux, Pierre

    2016-04-01

    Metabolic adaptations are intimately associated with changes in cell behavior. Cancers are characterized by a high metabolic plasticity resulting from mutations and the selection of metabolic phenotypes conferring growth and invasive advantages. While metabolic plasticity allows cancer cells to cope with various microenvironmental situations that can be encountered in a primary tumor, there is increasing evidence that metabolism is also a major driver of cancer metastasis. Rather than a general switch promoting metastasis as a whole, a succession of metabolic adaptations is more likely needed to promote different steps of the metastatic process. This review addresses the contribution of pH, glycolysis and the pentose phosphate pathway, and a companion paper summarizes current knowledge regarding the contribution of mitochondria, lipids and amino acid metabolism. Extracellular acidification, intracellular alkalinization, the glycolytic enzyme phosphoglucose isomerase acting as an autocrine cytokine, lactate and the pentose phosphate pathway are emerging as important factors controlling cancer metastasis.

  6. Average snowcover density values in Eastern Alps mountain

    NASA Astrophysics Data System (ADS)

    Valt, M.; Moro, D.

    2009-04-01

    The Italian Avalanche Warning Services monitor the snow cover characteristics through networks evenly distributed all over the alpine chain. Measurements of snow stratigraphy and density are very frequently performed with sampling rates of 1 -2 times per week. Snow cover density values are used to compute the dimensions of the building roofs as well as to design avalanche barriers. Based on the measured snow densities the Electricity Board can predict the amount of water resources deriving from snow melt in high relieves drainage basins. In this work it was possible to compute characteristic density values of the snow cover in the Eastern Alps using the information contained in the database from the ARPA (Agenzia Regionale Protezione Ambiente)-Centro Valanghe di Arabba, and Ufficio Valanghe- Udine. Among the other things, this database includes 15 years of stratigraphic measurements. More than 6,000 snow stratigraphic logs were analysed, in order to derive typical values as for geographical area, altitude, exposure, snow cover thickness and season. Computed values were compared to those established by the current Italian laws. Eventually, experts identified and evaluated the correlations between the seasonal variations of the average snow density and the variations related to the snowfall rate in the period 1994-2008 in the Eastern Alps mountain range

  7. Migration and transformation of different phosphorus forms in rainfall runoff in bioretention system.

    PubMed

    Song, Yujia; Song, Shoufa

    2018-06-04

    Artificial bioretention system consisting of Ophiopogon japonicus infiltration medium was used to simulate an infiltration experiment of rainfall runoff. Continuous extraction method was used to detect contents of inorganic phosphorus (P) under exchangeable state (Ex-P) and aluminium phosphate (Al-P) and iron phosphate (Fe-P) at different depths (0, 5, 15 and 35 cm) of soil infiltration medium in bioretention system. Effluent total P (TP) concentration of the system was also monitored. Results indicated that the adsorption of inorganic P, Al-P and Fe-P by soil infiltration medium was implemented layer by layer from top to bottom and gradually weakened. Moreover, Ex-P was gradually transformed into Al-P and Fe-P, whereas Al-P was gradually transformed into Fe-P; thus, Ex-P content reduced layer by layer, whereas Al-P and Fe-P gradually accumulated. The TP removal rate in runoff rainwater by the system was more than 90%, where the TP that was not used by plants was under dynamic equilibrium in water-soil-root system/biological system.

  8. The effect of reaction conditions on formation of wet precipitated calcium phosphates

    NASA Astrophysics Data System (ADS)

    Huang, Chen; Cao, Peng

    2015-03-01

    The precipitation process discussed in the present study involves the addition of alkaline solutions to an acidic calcium phosphate suspension. Several parameters (pH, pH buffer reagent, ageing and stirring) were investigated. The synthesized powders were calcined at 1000°C for 1 h in air, in order to study the thermal stability and crystalline phase compositions. X-ray diffraction (XRD) and ESEM analysis were used for sample characterization. It is found that all these processing parameters affect the crystalline phases evolved and resultant microstructures. Phase evolution occurred at an elevated pH level. The pH buffer reagent would affect both the phase composition and microstructure. Ageing was essential for the phase maturation. Stirring accelerated the reaction process by providing a homogeneous medium for precipitation.

  9. Quinone Methide Signal Amplification: Covalent Reporter Labeling of Cancer Epitopes using Alkaline Phosphatase Substrates.

    PubMed

    Polaske, Nathan W; Kelly, Brian D; Ashworth-Sharpe, Julia; Bieniarz, Christopher

    2016-03-16

    Diagnostic assays with the sensitivity required to improve cancer therapeutics depend on the development of new signal amplification technologies. Herein, we report the development and application of a novel amplification system which utilizes latent quinone methides (QMs) activated by alkaline phosphatase (AP) for signal amplification in solid-phase immunohistochemical (IHC) assays. Phosphate-protected QM precursor substrates were prepared and conjugated to either biotin or a fluorophore through an amine-functionalized linker group. Upon reaction with AP, the phosphate group is cleaved, followed by elimination of the leaving group and formation of the highly reactive and short-lived QM. The QMs either react with tissue nucleophiles in close proximity to their site of generation, or are quenched by nucleophiles in the reaction media. The reporter molecules that covalently bind to the tissue were then detected visually by fluorescence microscopy in the case of fluorophore reporters, or brightfield microscopy using diaminobenzidine (DAB) in the case of biotin reporters. With multiple reporters deposited per enzyme, significant signal amplification was observed utilizing QM precursor substrates containing either benzyl difluoro or benzyl monofluoro leaving group functionalities. However, the benzyl monofluoro leaving group gave superior results with respect to both signal intensity and discretion, the latter of which was found to be imperative for use in diagnostic IHC assays.

  10. Ice cap melting and low viscosity crustal root explain narrow geodetic uplift of the Western Alps

    NASA Astrophysics Data System (ADS)

    Chery, Jean; Genti, Manon; Vernant, Philippe

    2016-04-01

    More than 10 years of geodetic measurements demonstrate an uplift rate of 1-3 mm/yr of the high topography region of the Western Alps. By contrast, no significant horizontal motion has been detected. Three uplift mechanisms have been proposed so far: (1) the isostatic response to denudation. However this process is responsible for only a fraction of the observed uplift and (2) the rebound induced by the Wurmian ice cap melting. This process leads to a broader uplifting region than the one evidenced by geodetic observations. (3) a deep source motion associated with slab motion or some deep isostatic unbalance. Using a numerical model accounting for crustal and mantle rheology of the Alps and its foreland, we model the response to Wurmian ice cap melting. We show that a crustal viscosity contrast between the foreland and the central part of the Alps, the later being weaker with a viscosity of 1021 Pa.s, is needed to produce a narrow uplift. The vertical rates are enhanced if the strong uppermost mantle beneath the Moho is interrupted across the Alps, therefore allowing a weak vertical rheological anomaly thanks to the continuity between the low viscosity parts of the crust and mantle. References: Champagnac, J.-D., F. Schlunegger, K. Norton, F. von Blanckenburg, L. M. Abbühl, and M. Schwab (2009), Erosion-driven uplift of the modern Central Alps, Tectonophysics, 474(1-2), 236-249. Vernant, P., F. Hivert, J. Chéry, P. Steer, R. Cattin, and A. Rigo (2013), Erosion-induced isostatic rebound triggers extension in low convergent mountain ranges, geology, 41(4), 467-470.

  11. Net alkalinity and net acidity 2: Practical considerations

    USGS Publications Warehouse

    Kirby, C.S.; Cravotta, C.A.

    2005-01-01

    The pH, alkalinity, and acidity of mine drainage and associated waters can be misinterpreted because of the chemical instability of samples and possible misunderstandings of standard analytical method results. Synthetic and field samples of mine drainage having various initial pH values and concentrations of dissolved metals and alkalinity were titrated by several methods, and the results were compared to alkalinity and acidity calculated based on dissolved solutes. The pH, alkalinity, and acidity were compared between fresh, unoxidized and aged, oxidized samples. Data for Pennsylvania coal mine drainage indicates that the pH of fresh samples was predominantly acidic (pH 2.5-4) or near neutral (pH 6-7); ??? 25% of the samples had pH values between 5 and 6. Following oxidation, no samples had pH values between 5 and 6. The Standard Method Alkalinity titration is constrained to yield values >0. Most calculated and measured alkalinities for samples with positive alkalinities were in close agreement. However, for low-pH samples, the calculated alkalinity can be negative due to negative contributions by dissolved metals that may oxidize and hydrolyze. The Standard Method hot peroxide treatment titration for acidity determination (Hot Acidity) accurately indicates the potential for pH to decrease to acidic values after complete degassing of CO2 and oxidation of Fe and Mn, and it indicates either the excess alkalinity or that required for neutralization of the sample. The Hot Acidity directly measures net acidity (= -net alkalinity). Samples that had near-neutral pH after oxidation had negative Hot Acidity; samples that had pH < 6.3 after oxidation had positive Hot Acidity. Samples with similar pH values before oxidation had dissimilar Hot Acidities due to variations in their alkalinities and dissolved Fe, Mn, and Al concentrations. Hot Acidity was approximately equal to net acidity calculated based on initial pH and dissolved concentrations of Fe, Mn, and Al minus the

  12. Evaluation of Alkaline Cleaner Materials

    NASA Technical Reports Server (NTRS)

    Partz, Earl

    1998-01-01

    Alkaline cleaners used to process aluminum substrates have contained chromium as the corrosion inhibitor. Chromium is a hazardous substance whose use and control are described by environmental laws. Replacement materials that have the characteristics of chromated alkaline cleaners need to be found that address both the cleaning requirements and environmental impacts. This report will review environmentally friendly candidates evaluated as non-chromium alkaline cleaner replacements and methods used to compare those candidates one versus another. The report will also list characteristics used to select candidates based on their declared contents. It will also describe and evaluate methods used to discriminate among the large number of prospective candidates.

  13. Coregulated Expression of the Na+/Phosphate Pho89 Transporter and Ena1 Na+-ATPase Allows Their Functional Coupling under High-pH Stress

    PubMed Central

    Serra-Cardona, Albert; Petrezsélyová, Silvia; Canadell, David; Ramos, José

    2014-01-01

    The yeast Saccharomyces cerevisiae has two main high-affinity inorganic phosphate (Pi) transporters, Pho84 and Pho89, that are functionally relevant at acidic/neutral pH and alkaline pH, respectively. Upon Pi starvation, PHO84 and PHO89 are induced by the activation of the PHO regulon by the binding of the Pho4 transcription factor to specific promoter sequences. We show that PHO89 and PHO84 are induced by alkalinization of the medium with different kinetics and that the network controlling Pho89 expression in response to alkaline pH differs from that of other members of the PHO regulon. In addition to Pho4, the PHO89 promoter is regulated by the transcriptional activator Crz1 through the calcium-activated phosphatase calcineurin, and it is under the control of several repressors (Mig2, Nrg1, and Nrg2) coordinately regulated by the Snf1 protein kinase and the Rim101 transcription factor. This network mimics the one regulating expression of the Na+-ATPase gene ENA1, encoding a major determinant for Na+ detoxification. Our data highlight a scenario in which the activities of Pho89 and Ena1 are functionally coordinated to sustain growth in an alkaline environment. PMID:25266663

  14. Acid Gradient across Plasma Membrane Can Drive Phosphate Bond Synthesis in Cancer Cells: Acidic Tumor Milieu as a Potential Energy Source

    PubMed Central

    Dhar, Gautam; Sen, Suvajit; Chaudhuri, Gautam

    2015-01-01

    Aggressive cancers exhibit an efficient conversion of high amounts of glucose to lactate accompanied by acid secretion, a phenomenon popularly known as the Warburg effect. The acidic microenvironment and the alkaline cytosol create a proton-gradient (acid gradient) across the plasma membrane that represents proton-motive energy. Increasing experimental data from physiological relevant models suggest that acid gradient stimulates tumor proliferation, and can also support its energy needs. However, direct biochemical evidence linking extracellular acid gradient to generation of intracellular ATP are missing. In this work, we demonstrate that cancer cells can synthesize significant amounts of phosphate-bonds from phosphate in response to acid gradient across plasma membrane. The noted phenomenon exists in absence of glycolysis and mitochondrial ATP synthesis, and is unique to cancer. Biochemical assays using viable cancer cells, and purified plasma membrane vesicles utilizing radioactive phosphate, confirmed phosphate-bond synthesis from free phosphate (Pi), and also localization of this activity to the plasma membrane. In addition to ATP, predominant formation of pyrophosphate (PPi) from Pi was also observed when plasma membrane vesicles from cancer cells were subjected to trans-membrane acid gradient. Cancer cytosols were found capable of converting PPi to ATP, and also stimulate ATP synthesis from Pi from the vesicles. Acid gradient created through glucose metabolism by cancer cells, as observed in tumors, also proved critical for phosphate-bond synthesis. In brief, these observations reveal a role of acidic tumor milieu as a potential energy source and may offer a novel therapeutic target. PMID:25874623

  15. Variation trend of snowfall in the Kamikochi region of the Japanese Alps

    NASA Astrophysics Data System (ADS)

    Suzuki, K.

    2017-12-01

    The Japanese Alps experience exceptionally heavy snowfall, extreme even by global standards, and in spring and summer the melting snow becomes a valuable water resource. The snow effectively acts as a natural dam when it accumulates in watersheds during winter. However, there have been no observations of the amount of snow in high-altitude regions of Japan. Therefore, we cannot discuss the effect of global warming on the change in the amount of snow in these regions based on direct observation data. We were, however, able to obtain climatic and hydrologic data for high-altitude sites in the Japanese Alps, and discuss the variations in these conditions in the Kamikochi region (altitude 1490 m-3190 m) of the Japanese Alps over a 68-year period using these observed data. No long-term trends are observed in the annual mean, maximum, or minimum temperatures at Taisho-ike from 1945 to 2012; the total annual precipitation shows a statistically significant decreasing trend. The annual total snowfall at Taisho-ike from 1969 to 2012 shows a statistically significant increasing trend. The annual total runoff of the Azusa River from 1945 to 2012 shows a statistically significant increasing trend, as does the snowmelt runoff to the river (which occurs from May to July). We can thus conclude that the annual snowfall in the Azusa River catchment has increased in recent years.

  16. Two Pore Channel 2 (TPC2) Inhibits Autophagosomal-Lysosomal Fusion by Alkalinizing Lysosomal pH*

    PubMed Central

    Lu, Yingying; Hao, Bai-Xia; Graeff, Richard; Wong, Connie W. M.; Wu, Wu-Tian; Yue, Jianbo

    2013-01-01

    Autophagy is an evolutionarily conserved lysosomal degradation pathway, yet the underlying mechanisms remain poorly understood. Nicotinic acid adenine dinucleotide phosphate (NAADP), one of the most potent Ca2+ mobilizing messengers, elicits Ca2+ release from lysosomes via the two pore channel 2 (TPC2) in many cell types. Here we found that overexpression of TPC2 in HeLa or mouse embryonic stem cells inhibited autophagosomal-lysosomal fusion, thereby resulting in the accumulation of autophagosomes. Treatment of TPC2 expressing cells with a cell permeant-NAADP agonist, NAADP-AM, further induced autophagosome accumulation. On the other hand, TPC2 knockdown or treatment of cells with Ned-19, a NAADP antagonist, markedly decreased the accumulation of autophagosomes. TPC2-induced accumulation of autophagosomes was also markedly blocked by ATG5 knockdown. Interestingly, inhibiting mTOR activity failed to increase TPC2-induced autophagosome accumulation. Instead, we found that overexpression of TPC2 alkalinized lysosomal pH, and lysosomal re-acidification abolished TPC2-induced autophagosome accumulation. In addition, TPC2 overexpression had no effect on general endosomal-lysosomal degradation but prevented the recruitment of Rab-7 to autophagosomes. Taken together, our data demonstrate that TPC2/NAADP/Ca2+ signaling alkalinizes lysosomal pH to specifically inhibit the later stage of basal autophagy progression. PMID:23836916

  17. Alkaline Phosphatases in the Complex Chronic Kidney Disease-Mineral and Bone Disorders.

    PubMed

    Bover, Jordi; Ureña, Pablo; Aguilar, Armando; Mazzaferro, Sandro; Benito, Silvia; López-Báez, Víctor; Ramos, Alejandra; daSilva, Iara; Cozzolino, Mario

    2018-02-14

    Alkaline phosphatases (APs) remove the phosphate (dephosphorylation) needed in multiple metabolic processes (from many molecules such as proteins, nucleotides, or pyrophosphate). Therefore, APs are important for bone mineralization but paradoxically they can also be deleterious for other processes, such as vascular calcification and the increasingly known cross-talk between bone and vessels. A proper balance between beneficial and harmful activities is further complicated in the context of chronic kidney disease (CKD). In this narrative review, we will briefly update the complexity of the enzyme, including its different isoforms such as the bone-specific alkaline phosphatase or the most recently discovered B1x. We will also analyze the correlations and potential discrepancies with parathyroid hormone and bone turnover and, most importantly, the valuable recent associations of AP's with cardiovascular disease and/or vascular calcification, and survival. Finally, a basic knowledge of the synthetic and degradation pathways of APs promises to open new therapeutic strategies for the treatment of the CKD-Mineral and Bone Disorder (CKD-MBD) in the near future, as well as for other processes such as sepsis, acute kidney injury, inflammation, endothelial dysfunction, metabolic syndrome or, in diabetes, cardiovascular complications. However, no studies have been done using APs as a primary therapeutic target for clinical outcomes, and therefore, AP's levels cannot yet be used alone as an isolated primary target in the treatment of CKD-MBD. Nonetheless, its diagnostic and prognostic potential should be underlined.

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

    PubMed Central

    2016-01-01

    Naively one might have expected an early division between phosphate monoesterases and diesterases of the alkaline phosphatase (AP) superfamily. On the contrary, prior results and our structural and biochemical analyses of phosphate monoesterase PafA, from Chryseobacterium meningosepticum, indicate similarities to a superfamily phosphate diesterase [Xanthomonas citri nucleotide pyrophosphatase/phosphodiesterase (NPP)] and distinct differences from the three metal ion AP superfamily monoesterase, from Escherichia coli AP (EcAP). We carried out a series of experiments to map out and learn from the differences and similarities between these enzymes. First, we asked why there would be independent instances of monoesterases in the AP superfamily? PafA has a much weaker product inhibition and slightly higher activity relative to EcAP, suggesting that different metabolic evolutionary pressures favored distinct active-site architectures. Next, we addressed the preferential phosphate monoester and diester catalysis of PafA and NPP, respectively. We asked whether the >80% sequence differences throughout these scaffolds provide functional specialization for each enzyme’s cognate reaction. In contrast to expectations from this model, PafA and NPP mutants with the common subset of active-site groups embedded in each native scaffold had the same monoesterase:diesterase specificities; thus, the >107-fold difference in native specificities appears to arise from distinct interactions at a single phosphoryl substituent. We also uncovered striking mechanistic similarities between the PafA and EcAP monoesterases, including evidence for ground-state destabilization and functional active-site networks that involve different active-site groups but may play analogous catalytic roles. Discovering common network functions may reveal active-site architectural connections that are critical for function, and identifying regions of functional modularity may facilitate the design of new enzymes

  19. Intra-continental subduction and contemporaneous lateral extrusion of the upper plate: insights into Alps-Adria interactions

    NASA Astrophysics Data System (ADS)

    van Gelder, Inge; Willingshofer, Ernst; Sokoutis, Dimitrios; Cloetingh, Sierd

    2017-04-01

    A series of physical analogue experiments were performed to simulate intra-continental subduction contemporaneous with lateral extrusion of the upper plate to study the interferences between these two processes at crustal levels and in the lithospheric mantle. The lithospheric-scale models are specifically designed to represent the collision of the Adriatic microplate with the Eastern Alps, simulated by an intra-continental weak zone to initiate subduction and a weak confined margin perpendicular to the direction of convergence in order to allow for extrusion of the lithosphere. The weak confined margin is the analog for the opening of the Pannonian back-arc basin adjacent to the Eastern Alps with the direction of extension perpendicular to the strike of the orogen. The models show that intra-continental subduction and coeval lateral extrusion of the upper plate are compatible processes. The obtained deformation structures within the extruding region are similar compared to the classical setup where lateral extrusion is provoked by lithosphere-scale indentation. In the models a strong coupling across the subduction boundary allows for the transfer of abundant stresses to the upper plate, leading to laterally varying strain regimes that are characterized by crustal thickening near a confined margin and dominated by lateral displacement of material near a weak lateral confinement. During ongoing convergence the strain regimes propagate laterally, thereby creating an area of overlap characterized by transpression. In models with oblique subduction, with respect to the convergence direction, less deformation of the upper plate is observed and as a consequence the amount of lateral extrusion decreases. Additionally, strain is partitioned along the oblique plate boundary leading to less subduction in expense of right lateral displacement close to the weak lateral confinement. Both oblique and orthogonal subduction models have a strong resemblance to lateral extrusion

  20. Phosphate removal and hemodialysis conditions.

    PubMed

    Pohlmeier, R; Vienken, J

    2001-02-01

    Hyperphosphatemia is frequently found in hemodialysis patients, and the association with an increased risk of mortality has been demonstrated. Other authors have linked hyperphosphatemia to increased cardiovascular mortality. The normalization of phosphate plasma levels is therefore an important goal in the treatment of end-stage renal disease patients. Absorption of phosphate from the food exceeds the elimination through a hemodialysis treatment, and this leads to a chronic phosphate load for the majority of hemodialysis patients. This imbalance should be improved by either a reduction of phosphate absorption or an increased removal of phosphate. A reduction of phosphate absorption can be achieved by reducing the amount of phosphate in the diet or by the administration of phosphate binders. Unfortunately, these measures imply practical difficulties, for example, a lack of patient compliance or other side effects. When considering modifications of the hemodialysis treatment, an essential understanding of the kinetics of dialytic phosphate removal is mandatory. Phosphate is unevenly distributed in different compartments of the body. Only a very small amount of phosphate is present in the easily accessible plasma compartment. The major part of phosphate removed during hemodialysis originates from the cytoplasm of cells. A transfer from intracellular space to the plasma and further from the plasma to the dialysate is necessary. However, if we consider improvement to phosphate removal by dialysis procedures, full dialyzer clearance is effective in only the initial phase of the dialysis treatment. After this initial phase, the transfer rate for phosphate from the intracellular space to the plasma becomes the rate-limiting step for phosphate transport. Attempts to improve this transfer rate have recently been investigated by acidosis correction, but turned out not to be consistently successful. Furthermore, modifications of the treatment schedule have been described in

  1. Substantial differences between human and ovine mesenchymal stem cells in response to osteogenic media: how to explain and how to manage?

    PubMed

    Kalaszczynska, Ilona; Ruminski, Slawomir; Platek, Anna E; Bissenik, Igor; Zakrzewski, Piotr; Noszczyk, Maria; Lewandowska-Szumiel, Malgorzata

    2013-10-01

    It is expected that use of adult multipotential mesenchymal stem cells (MSCs) for bone tissue engineering (TE) will lead to improvement of TE products. Prior to clinical application, biocompatibility of bone TE products need to be tested in vitro and in vivo. In orthopedic research, sheep are a well-accepted model due to similarities with humans and are assumed to be predictive of human outcomes. In this study we uncover differences between human and ovine bone marrow-derived MSCs (BMSCs) and adipose tissue-derived MSCs (ADSCs) in response to osteogenic media. Osteogenic differentiation of BMSCs and ADSCs was monitored by alkaline phosphatase (ALP) activity and calcium deposition. Mineralization of ovine BMSC was achieved in medium containing NaH2PO4 as a source of phosphate ions (Pi), but not in medium containing β-glycerophosphate (β-GP), which is most often used. In a detailed study we found no induction of ALP activity in ovine BMSCs and ADSCs upon osteogenic stimulation, which makes β-GP an unsuitable source of phosphate ions for ovine cells. Moreover, mineralization of human ADSCs was more efficient in osteogenic medium containing NaH2PO4. These results indicate major differences between ovine and human MSCs and suggest that standard in vitro osteogenic differentiation techniques may not be suitable for all types of cells used in cell-based therapies. Since mineralization is a widely accepted marker of the osteogenic differentiation and maturation of cells in culture, it may lead to potentially misleading results and should be taken into account at the stage of planning and interpreting preclinical observations performed in animal models. We also present a cell culture protocol for ovine ADSCs, which do not express ALP activity and do not mineralize under routine pro-osteogenic conditions in vitro. We plan to apply it in preclinical experiments of bone tissue-engineered products performed in an ovine model.

  2. Substantial Differences Between Human and Ovine Mesenchymal Stem Cells in Response to Osteogenic Media: How to Explain and How to Manage?

    PubMed Central

    Kalaszczynska, Ilona; Ruminski, Slawomir; Platek, Anna E.; Bissenik, Igor; Zakrzewski, Piotr; Noszczyk, Maria

    2013-01-01

    Abstract It is expected that use of adult multipotential mesenchymal stem cells (MSCs) for bone tissue engineering (TE) will lead to improvement of TE products. Prior to clinical application, biocompatibility of bone TE products need to be tested in vitro and in vivo. In orthopedic research, sheep are a well-accepted model due to similarities with humans and are assumed to be predictive of human outcomes. In this study we uncover differences between human and ovine bone marrow–derived MSCs (BMSCs) and adipose tissue–derived MSCs (ADSCs) in response to osteogenic media. Osteogenic differentiation of BMSCs and ADSCs was monitored by alkaline phosphatase (ALP) activity and calcium deposition. Mineralization of ovine BMSC was achieved in medium containing NaH2PO4 as a source of phosphate ions (Pi), but not in medium containing β-glycerophosphate (β-GP), which is most often used. In a detailed study we found no induction of ALP activity in ovine BMSCs and ADSCs upon osteogenic stimulation, which makes β-GP an unsuitable source of phosphate ions for ovine cells. Moreover, mineralization of human ADSCs was more efficient in osteogenic medium containing NaH2PO4. These results indicate major differences between ovine and human MSCs and suggest that standard in vitro osteogenic differentiation techniques may not be suitable for all types of cells used in cell-based therapies. Since mineralization is a widely accepted marker of the osteogenic differentiation and maturation of cells in culture, it may lead to potentially misleading results and should be taken into account at the stage of planning and interpreting preclinical observations performed in animal models. We also present a cell culture protocol for ovine ADSCs, which do not express ALP activity and do not mineralize under routine pro-osteogenic conditions in vitro. We plan to apply it in preclinical experiments of bone tissue–engineered products performed in an ovine model. PMID:24083091

  3. Medical ethnobotany of the Albanian Alps in Kosovo

    PubMed Central

    2012-01-01

    Background Ethnobotanical studies are crucial in South-Eastern Europe for fostering local development and also for investigating the dynamics of Traditional Environmental Knowledge (TEK) related to plants in one of the most crucial European hotspots for biocultural diversity. The current medico-ethnobotanical survey was conducted in rural alpine communities in Kosovo. The aims of the study were twofold: 1) to document the state of TEK of medicinal plants in these communities; 2) to compare these findings with that of similar field studies previously conducted among local populations inhabiting the Montenegrin and Albanian side of the same Alpine range. Methods Field research was conducted in 36 villages on the Kosovar side of the Albanian Alps. Snowball sampling techniques were used to recruit 91 elderly informants (≥ 50 years-old) for participation in semi-structured interviews and structured surveys regarding the use of the local flora for medicinal and food purposes. Standard ethnobotanical methods were employed and prior informed consent was obtained for all study participants. Results and Conclusion The uses of 98 plants species belonging to 39 families were recorded; the most quoted botanical families were Rosaceae, Asteraceae, and Lamiaceae. Mainly decoctions and infusions were quoted as folk medicinal preparations and the most common uses referred to gastrointestinal and respiratory disorders, as well as illnesses of the uro-genital system. Among the most uncommon medicinal taxa quoted by the informants, Carduus nutans L., Echinops bannaticus Rochel ex Schrad., and Orlaya grandiflora Hoffm. may merit phytochemical and phytopharmacological investigations. Comparison of the data with other ethnobotanical field studies recently conducted on the Albanian and Montenegrin sides of the same Alps has shown a remarkable link between the medical ethnobotany of Montenegrin and Kosovar side of the Albanian Alps. Moreover, folk uses of the most quoted wild medicinal

  4. Phosphate-a poison for humans?

    PubMed

    Komaba, Hirotaka; Fukagawa, Masafumi

    2016-10-01

    Maintenance of phosphate balance is essential for life, and mammals have developed a sophisticated system to regulate phosphate homeostasis over the course of evolution. However, due to the dependence of phosphate elimination on the kidney, humans with decreased kidney function are likely to be in a positive phosphate balance. Phosphate excess has been well recognized as a critical factor in the pathogenesis of mineral and bone disorders associated with chronic kidney disease, but recent investigations have also uncovered toxic effects of phosphate on the cardiovascular system and the aging process. Compelling evidence also suggests that increased fibroblastic growth factor 23 and parathyroid hormone levels in response to a positive phosphate balance contribute to adverse clinical outcomes. These insights support the current practice of managing serum phosphate in patients with advanced chronic kidney disease, although definitive evidence of these effects is lacking. Given the potential toxicity of excess phosphate, the general population may also be viewed as a target for phosphate management. However, the widespread implementation of dietary phosphate intervention in the general population may not be warranted due to the limited impact of increased phosphate intake on mineral metabolism and clinical outcomes. Nonetheless, the increasing incidence of kidney disease or injury in our aging society emphasizes the potential importance of this issue. Further work is needed to more completely characterize phosphate toxicity and to establish the optimal therapeutic strategy for managing phosphate in patients with chronic kidney disease and in the general population. Copyright © 2016 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.

  5. Environmental Characteristics of Carbonatite and Alkaline Intrusion-related Rare Earth Element (REE) Deposits

    NASA Astrophysics Data System (ADS)

    Seal, R. R., II; Piatak, N. M.

    2017-12-01

    Carbonatites and alkaline intrusions are important sources of REEs. Environmental risks related to these deposit types have been assessed through literature review and evaluation of the geochemical properties of representative samples of mill tailings and their leachates. The main ore mineral in carbonatite deposits is bastnasite [(Ce,La)(CO3)F], which is found with dolomite and calcite ( 65 %), barite (20 - 25 %), plus a number of minor accessory minerals including sulfides such as galena and pyrite. Generally, alkaline intrusion-related REE deposits either occur in layered complexes or with dikes and veins cutting alkaline intrusions. Such intrusions have a more diverse group of REE ore minerals that include fluorcarbonates, oxides, silicates, and phosphates. Ore also can include minor calcite and iron (Fe), lead (Pb), and zinc (Zn) sulfides. The acid-generating potential of both deposit types is low because of a predominance of carbonate minerals in the carbonatite deposits, the presence of feldspars and minor calcite in alkaline intrusion-related deposits, and to only minor to trace occurrence of potentially acid-generating sulfide minerals. Both deposit types, however, are produced by igneous and hydrothermal processes that enrich high-field strength, incompatible elements, which typically are excluded from common rock-forming minerals. Elements such as yttrium (Y), niobium Nb), zirconium (Zr), hafnium (Hf), tungsten (W), titanium (Ti), tantalum (Ta), scandium (Sc), thorium (Th), and uranium (U) can be characteristic of these deposits and may be of environmental concern. Most of these elements, including the REEs, but with the exception of U, have low solubilities in water at the near-neutral pH values expected around these deposits. Mill tailings from carbonatite deposits can exceed residential soil and sediment criteria for Pb, and leachates from mill tailings can exceed drinking water guidelines for Pb. The greatest environmental challenges, however, are

  6. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    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,more » 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.« less

  7. Cretaceous to Tertiary paleogeographic reconstructions of the Alps-Pyrenees linking zone

    NASA Astrophysics Data System (ADS)

    Frasca, Gianluca; Dielforder, Armin; Ford, Mary; Vergés, Jaume

    2017-04-01

    The northwestern Mediterranean subduction systems underwent an important phase of reorganization between Late Cretaceous and Eocene. The mode and timing of this reorganization are still under debate. Great uncertainties mainly derive from the poorly preserved record of the early phases of orogenic evolution in both the Alps and Pyrenees and the distruction of the orogenic system between the Pyrenees and Alps by the Oligo-Miocene opening of the Gulf of Lion due to backarc rifting. Vestiges are nevertheless preserved in the Pyreneo-Provençal fold-and-thrust belt and associated basins in southern France and Corsica-Sardinia. In this work we first review published plate kinematic models for Iberia, Apulia and Europe from 83 Ma, focusing in particular on the restoration of the Corso-Sardinia block using the free software GPlates. Second, we characterize the Upper Cretaceous to Eocene depositional systems at the junction between the Alps, Pyrenees and Apennines, reviewing previous paleogeographic restorations for the Western Alpine and Eastern Pyrenean foreland basins. Last, we compare the kinematic models with reconstructed basin dynamics. We critically assess the implications of newly proposed paleogeographic reconstructions (at 83, 65, 50, 37 and 30 Ma) for the validity of various plate kinematic models. The information derived from the sedimentary basins help to define the mode and timing of the subduction reorganization that occurred between 83 and 30 Ma in the northwestern Mediterranean. This study is part of the Orogen research program funded by Total, the BRGM (Bureau de Recherches Géologiques et Minières), the CNRS (Centre National de la Recherche Scientifique).

  8. Increased river alkalinization in the Eastern U.S.

    PubMed

    Kaushal, Sujay S; Likens, Gene E; Utz, Ryan M; Pace, Michael L; Grese, Melissa; Yepsen, Metthea

    2013-09-17

    The interaction between human activities and watershed geology is accelerating long-term changes in the carbon cycle of rivers. We evaluated changes in bicarbonate alkalinity, a product of chemical weathering, and tested for long-term trends at 97 sites in the eastern United States draining over 260,000 km(2). We observed statistically significant increasing trends in alkalinity at 62 of the 97 sites, while remaining sites exhibited no significant decreasing trends. Over 50% of study sites also had statistically significant increasing trends in concentrations of calcium (another product of chemical weathering) where data were available. River alkalinization rates were significantly related to watershed carbonate lithology, acid deposition, and topography. These three variables explained ~40% of variation in river alkalinization rates. The strongest predictor of river alkalinization rates was carbonate lithology. The most rapid rates of river alkalinization occurred at sites with highest inputs of acid deposition and highest elevation. The rise of alkalinity in many rivers throughout the Eastern U.S. suggests human-accelerated chemical weathering, in addition to previously documented impacts of mining and land use. Increased river alkalinization has major environmental implications including impacts on water hardness and salinization of drinking water, alterations of air-water exchange of CO2, coastal ocean acidification, and the influence of bicarbonate availability on primary production.

  9. Phosphate and calcium are required for TGFbeta-mediated stimulation of ANK expression and function during chondrogenesis.

    PubMed

    Oca, Paulina; Zaka, Raihana; Dion, Arnold S; Freeman, Theresa A; Williams, Charlene J

    2010-08-01

    The expression of ANK, a key player in biomineralization, is stimulated by treatment with TGFbeta. The purpose of this study was to determine whether TGFbeta stimulation of ANK expression during chondrogenesis was dependent upon the influx of calcium and phosphate into cells. Treatment of ATDC5 cells with TGFbeta increased ANK expression during all phases of chondrogenic differentiation, particularly at day 14 (proliferation) and day 32 (mineralizing hypertrophy) of culture. Phosphate uptake studies in the presence and absence of phosphonoformic acid (PFA), a competitive inhibitor of the type III Na(+)/Pi channels Pit-1 and Pit-2, indicated that the stimulation of ANK expression by TGFbeta required the influx of phosphate, specifically by the Pit-1 transporter, at all phases of differentiation. At hypertrophy, when alkaline phosphatase is highly expressed, inhibition of its activity with levamisole also abrogated the stimulatory effect of TGFbeta on ANK expression, further illustrating that Pi availability and uptake by the cells is necessary for stimulation of ANK expression in response to TGFbeta. Since previous studies of endochondral ossification in the growth plate have shown that L-type calcium channels are essential for chondrogenesis, we investigated their role in the TGFbeta-stimulated ANK response in ATDC5 cells. Treatment with nifedipine to inhibit calcium influx via the L-type channel Cav1.2 (alpha(1C)) inhibited the TGFbeta stimulated increase in ANK expression at all phases of chondrogenesis. Our findings indicate that TGFbeta stimulation of ANK expression is dependent upon the influx of phosphate and calcium into ATDC5 cells at all stages of differentiation.

  10. Osteogenic differentiation of equine adipose tissue derived mesenchymal stem cells using CaCl2.

    PubMed

    Elashry, Mohamed I; Baulig, Nadine; Heimann, Manuela; Bernhardt, Caroline; Wenisch, Sabine; Arnhold, Stefan

    2018-04-01

    Adipose tissue derived mesenchymal stem cells (ASCs) may be used to cure bone defects after osteogenic differentiation. In this study we tried to optimize osteogenic differentiation for equine ASCs using various concentrations of CaCl 2 in comparison to the standard osteogenic protocol. ASCs were isolated from subcutaneous adipose tissue from mixed breed horses. The osteogenic induction protocols were (1) the standard osteogenic medium (OM) composed of dexamethasone, ascorbic acid and β-glycerol phosphate; (2) CaCl 2 based protocol composed of 3, 5 and 7.5mM CaCl 2 . Differentiation and proliferation were evaluated at 7, 10, 14 and 21days post-differentiation induction using the alizarin red staining (ARS) detecting matrix calcification. Semi-quantification of cell protein content, ARS and alkaline phosphatase activity (ALP) were performed using an ELISA reader. Quantification of the transcription level for the common osteogenic markers alkaline phosphatase (ALP) and Osteopontin (OP) was performed using RT-qPCR. In the presence of CaCl 2 , a concentration dependent effect on the osteogenic differentiation capacity was evident by the ARS evaluation and OP gene expression. We provide evidence that 5 and 7mM CaCl 2 enhance the osteogenic differentiation compared to the OM protocol. Although, there was a clear commitment of ASCs to the osteogenic fate in the presence of 5 and 7mM CaCl 2 , cell proliferation was increased compared to OM. We report that an optimized CaCl 2 protocol reliably influences ASCs osteogenesis while conserving the proliferation capacity. Thus, using these protocols provide a platform for using ASCs as a cell source in bone tissue engineering. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Coupling alkaline pre-extraction with alkaline-oxidative post-treatment of corn stover to enhance enzymatic hydrolysis and fermentability.

    PubMed

    Liu, Tongjun; Williams, Daniel L; Pattathil, Sivakumar; Li, Muyang; Hahn, Michael G; Hodge, David B

    2014-04-03

    A two-stage chemical pretreatment of corn stover is investigated comprising an NaOH pre-extraction followed by an alkaline hydrogen peroxide (AHP) post-treatment. We propose that conventional one-stage AHP pretreatment can be improved using alkaline pre-extraction, which requires significantly less H2O2 and NaOH. To better understand the potential of this approach, this study investigates several components of this process including alkaline pre-extraction, alkaline and alkaline-oxidative post-treatment, fermentation, and the composition of alkali extracts. Mild NaOH pre-extraction of corn stover uses less than 0.1 g NaOH per g corn stover at 80°C. The resulting substrates were highly digestible by cellulolytic enzymes at relatively low enzyme loadings and had a strong susceptibility to drying-induced hydrolysis yield losses. Alkaline pre-extraction was highly selective for lignin removal over xylan removal; xylan removal was relatively minimal (~20%). During alkaline pre-extraction, up to 0.10 g of alkali was consumed per g of corn stover. AHP post-treatment at low oxidant loading (25 mg H2O2 per g pre-extracted biomass) increased glucose hydrolysis yields by 5%, which approached near-theoretical yields. ELISA screening of alkali pre-extraction liquors and the AHP post-treatment liquors demonstrated that xyloglucan and β-glucans likely remained tightly bound in the biomass whereas the majority of the soluble polymeric xylans were glucurono (arabino) xylans and potentially homoxylans. Pectic polysaccharides were depleted in the AHP post-treatment liquor relative to the alkaline pre-extraction liquor. Because the already-low inhibitor content was further decreased in the alkaline pre-extraction, the hydrolysates generated by this two-stage pretreatment were highly fermentable by Saccharomyces cerevisiae strains that were metabolically engineered and evolved for xylose fermentation. This work demonstrates that this two-stage pretreatment process is well suited for

  12. Coupling alkaline pre-extraction with alkaline-oxidative post-treatment of corn stover to enhance enzymatic hydrolysis and fermentability

    PubMed Central

    2014-01-01

    Background A two-stage chemical pretreatment of corn stover is investigated comprising an NaOH pre-extraction followed by an alkaline hydrogen peroxide (AHP) post-treatment. We propose that conventional one-stage AHP pretreatment can be improved using alkaline pre-extraction, which requires significantly less H2O2 and NaOH. To better understand the potential of this approach, this study investigates several components of this process including alkaline pre-extraction, alkaline and alkaline-oxidative post-treatment, fermentation, and the composition of alkali extracts. Results Mild NaOH pre-extraction of corn stover uses less than 0.1 g NaOH per g corn stover at 80°C. The resulting substrates were highly digestible by cellulolytic enzymes at relatively low enzyme loadings and had a strong susceptibility to drying-induced hydrolysis yield losses. Alkaline pre-extraction was highly selective for lignin removal over xylan removal; xylan removal was relatively minimal (~20%). During alkaline pre-extraction, up to 0.10 g of alkali was consumed per g of corn stover. AHP post-treatment at low oxidant loading (25 mg H2O2 per g pre-extracted biomass) increased glucose hydrolysis yields by 5%, which approached near-theoretical yields. ELISA screening of alkali pre-extraction liquors and the AHP post-treatment liquors demonstrated that xyloglucan and β-glucans likely remained tightly bound in the biomass whereas the majority of the soluble polymeric xylans were glucurono (arabino) xylans and potentially homoxylans. Pectic polysaccharides were depleted in the AHP post-treatment liquor relative to the alkaline pre-extraction liquor. Because the already-low inhibitor content was further decreased in the alkaline pre-extraction, the hydrolysates generated by this two-stage pretreatment were highly fermentable by Saccharomyces cerevisiae strains that were metabolically engineered and evolved for xylose fermentation. Conclusions This work demonstrates that this two

  13. Statistical downscaling of regional climate scenarios for the French Alps : Impacts on snow cover

    NASA Astrophysics Data System (ADS)

    Rousselot, M.; Durand, Y.; Giraud, G.; Mérindol, L.; Déqué, M.; Sanchez, E.; Pagé, C.; Hasan, A.

    2010-12-01

    Mountain areas are particularly vulnerable to climate change. Owing to the complexity of mountain terrain, climate research at scales relevant for impacts studies and decisive for stakeholders is challenging. A possible way to bridge the gap between these fine scales and those of the general circulation models (GCMs) consists of combining high-resolution simulations of Regional Climate Models (RCMs) to statistical downscaling methods. The present work is based on such an approach. It aims at investigating the impacts of climate change on snow cover in the French Alps for the periods 2021-2050 and 2071-2100 under several IPCC hypotheses. An analogue method based on high resolution atmospheric fields from various RCMs and climate reanalyses is used to simulate local climate scenarios. These scenarios, which provide meteorological parameters relevant for snowpack evolution, subsequently feed the CROCUS snow model. In these simulations, various sources of uncertainties are thus considered (several greenhouse gases emission scenarios and RCMs). Results are obtained for different regions of the French Alps at various altitudes. For all scenarios, temperature increase is relatively uniform over the Alps. This regional warming is larger than that generally modeled at the global scale (IPCC, 2007), and particularly strong in summer. Annual precipitation amounts seem to decrease, mainly as a result of decreasing precipitation trends in summer and fall. As a result of these climatic evolutions, there is a general decrease of the mean winter snow depth and seasonal snow duration for all massifs. Winter snow depths are particularly reduced in the Northern Alps. However, the impact on seasonal snow duration is more significant in the Southern and Extreme Southern Alps, since these regions are already characterized by small winter snow depths at low elevations. Reference : IPCC (2007a). Climate change 2007 : The physical science basis. Contribution of working group I to the

  14. Quality specifications for the extra-analytical phase of laboratory testing: Reference intervals and decision limits.

    PubMed

    Ceriotti, Ferruccio

    2017-07-01

    Reference intervals and decision limits are a critical part of the clinical laboratory report. The evaluation of their correct use represents a tool to verify the post analytical quality. Four elements are identified as indicators. 1. The use of decision limits for lipids and glycated hemoglobin. 2. The use, whenever possible, of common reference values. 3. The presence of gender-related reference intervals for at least the following common serum measurands (besides obviously the fertility relate hormones): alkaline phosphatase (ALP), alanine aminotransferase (ALT), creatine kinase (CK), creatinine, gamma-glutamyl transferase (GGT), IgM, ferritin, iron, transferrin, urate, red blood cells (RBC), hemoglobin (Hb) and hematocrit (Hct). 4. The presence of age-related reference intervals. The problem of specific reference intervals for elderly people is discussed, but their use is not recommended; on the contrary it is necessary the presence of pediatric age-related reference intervals at least for the following common serum measurands: ALP, amylase, creatinine, inorganic phosphate, lactate dehydrogenase, aspartate aminotransferase, urate, insulin like growth factor 1, white blood cells, RBC, Hb, Hct, alfa-fetoprotein and fertility related hormones. The lack of such reference intervals may imply significant risks for the patients. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  15. Effects of titanium surface anodization with CaP incorporation on human osteoblastic response

    PubMed Central

    OLIVEIRA, Natássia Cristina Martins; MOURA, Camilla Christian Gomes; ZANETTA-BARBOSA, Darceny; MENDONÇA, Daniela Baccelli Silveira; MENDONÇA, Gustavo; DECHICHI, Paula

    2015-01-01

    In this study we investigated whether anodization with calcium phosphate (CaP) incorporation (Vulcano®) enhances growth factors secretion, osteoblast-specific gene expression, and cell viability, when compared to acid etched surfaces (Porous®) and machined surfaces (Screw®) after 3 and 7 days. Results showed significant cell viability for Porous and Vulcano at day 7, when compared with Screw (p=0.005). At the same time point, significant differences regarding runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and bone sialoprotein (BSP) expression were found for all surfaces (p<0.05), but with greater fold induction for Porous and Vulcano. The secretion of transforming growth factor β1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) was not significantly affected by surface treatment in any experimental time (p>0.05). Although no significant correlation was found for growth factors secretion and Runx2 expression, a significant positive correlation between this gene and ALP/BSP expression showed that their strong association is independent on the type of surface. The incorporation of CaP affected the biological parameters evaluated similar to surfaces just acid etched. The results presented here support the observations that roughness also may play an important role in determining cell response. PMID:23498218

  16. ALPS: the Dark Matter Generator (coming in 2019)

    NASA Astrophysics Data System (ADS)

    Barke, Simon; Bush, Zachary; Baum, Claire; Hollis, Hal; Mueller, Guido; Tanner, David

    2017-01-01

    Very promising dark matter candidates are axion-like particles: sub-eV particles that are expected to (weakly) interact with photons in the presence of a static electric or magnetic field. This interaction can turn photons into axions and back into photons. Hence, in order to generate axions, we will set up a 100 meter long Fabry-Perot cavity that can hold a 150,000 watt laser field and have a 5.3 tesla magnetic field along the entire length. If the theory holds up, a fraction of the photons should transform into relativistic axions. These axions would then propagate through any optical barrier and enter a matched cavity that is situated within an identical magnetic field. Here, some of the axions should turn back into photons of equal energy. Thus these photons resonate in the otherwise empty cavity where they can be detected. It is unknown if axion-like particles exist in the targeted mass range. However, the ALPS detection principle is very convenient because we will know the exact energy of the regenerated photons beforehand thus making a detection much easier.The final stage of the ALPS experiment will be completed by 2019 at the German Electron Synchrotron (DESY) site in Hamburg, Germany. This work is supported by grants from the Heising-Simons Foundation and the National Science Foundation.

  17. Using phosphate supplementation to reverse hypophosphatemia and phosphate depletion in neurological disease and disturbance.

    PubMed

    Håglin, Lena

    2016-06-01

    Hypophosphatemia (HP) with or without intracellular depletion of inorganic phosphate (Pi) and adenosine triphosphate has been associated with central and peripheral nervous system complications and can be observed in various diseases and conditions related to respiratory alkalosis, alcoholism (alcohol withdrawal), diabetic ketoacidosis, malnutrition, obesity, and parenteral and enteral nutrition. In addition, HP may explain serious muscular, neurological, and haematological disorders and may cause peripheral neuropathy with paresthesias and metabolic encephalopathy, resulting in confusion and seizures. The neuropathy may be improved quickly after proper phosphate replacement. Phosphate depletion has been corrected using potassium-phosphate infusion, a treatment that can restore consciousness. In severe ataxia and tetra paresis, complete recovery can occur after adequate replacement of phosphate. Patients with multiple risk factors, often with a chronic disease and severe HP that contribute to phosphate depletion, are at risk for neurologic alterations. To predict both risk and optimal phosphate replenishment requires assessing the nutritional status and risk for re-feeding hypophosphatemia. The strategy for correcting HP depends on the severity of the underlying disease and the goal for re-establishing a phosphate balance to limit the consequences of phosphate depletion.

  18. Identification and characterization of miRNAs and targets in flax (Linum usitatissimum) under saline, alkaline, and saline-alkaline stresses.

    PubMed

    Yu, Ying; Wu, Guangwen; Yuan, Hongmei; Cheng, Lili; Zhao, Dongsheng; Huang, Wengong; Zhang, Shuquan; Zhang, Liguo; Chen, Hongyu; Zhang, Jian; Guan, Fengzhi

    2016-05-27

    MicroRNAs (miRNAs) play a critical role in responses to biotic and abiotic stress and have been characterized in a large number of plant species. Although flax (Linum usitatissimum L.) is one of the most important fiber and oil crops worldwide, no reports have been published describing flax miRNAs (Lus-miRNAs) induced in response to saline, alkaline, and saline-alkaline stresses. In this work, combined small RNA and degradome deep sequencing was used to analyze flax libraries constructed after alkaline-salt stress (AS2), neutral salt stress (NSS), alkaline stress (AS), and the non-stressed control (CK). From the CK, AS, AS2, and NSS libraries, a total of 118, 119, 122, and 120 known Lus-miRNAs and 233, 213, 211, and 212 novel Lus-miRNAs were isolated, respectively. After assessment of differential expression profiles, 17 known Lus-miRNAs and 36 novel Lus-miRNAs were selected and used to predict putative target genes. Gene ontology term enrichment analysis revealed target genes that were involved in responses to stimuli, including signaling and catalytic activity. Eight Lus-miRNAs were selected for analysis using qRT-PCR to confirm the accuracy and reliability of the miRNA-seq results. The qRT-PCR results showed that changes in stress-induced expression profiles of these miRNAs mirrored expression trends observed using miRNA-seq. Degradome sequencing and transcriptome profiling showed that expression of 29 miRNA-target pairs displayed inverse expression patterns under saline, alkaline, and saline-alkaline stresses. From the target prediction analysis, the miR398a-targeted gene codes for a copper/zinc superoxide dismutase, and the miR530 has been shown to explicitly target WRKY family transcription factors, which suggesting that these two micRNAs and their targets may significant involve in the saline, alkaline, and saline-alkaline stress response in flax. Identification and characterization of flax miRNAs, their target genes, functional annotations, and gene

  19. Biomediated continuous release phosphate fertilizer

    DOEpatents

    Goldstein, A.H.; Rogers, R.D.

    1999-06-15

    A composition is disclosed for providing phosphate fertilizer to the root zone of plants. The composition comprises a microorganism capable of producing and secreting a solubilization agent, a carbon source for providing raw material for the microorganism to convert into the solubilization agent, and rock phosphate ore for providing a source of insoluble phosphate that is solubilized by the solubilization agent and released as soluble phosphate. The composition is provided in a physical form, such as a granule, that retains the microorganism, carbon source, and rock phosphate ore, but permits water and soluble phosphate to diffuse into the soil. A method of using the composition for providing phosphate fertilizer to plants is also disclosed. 13 figs.

  20. Biomediated continuous release phosphate fertilizer

    DOEpatents

    Goldstein, Alan H.; Rogers, Robert D.

    1999-01-01

    A composition is disclosed for providing phosphate fertilizer to the root zone of plants. The composition comprises a microorganism capable of producing and secreting a solubilization agent, a carbon source for providing raw material for the microorganism to convert into the solubilization agent, and rock phosphate ore for providing a source of insoluble phosphate that is solubilized by the solubilization agent and released as soluble phosphate. The composition is provided in a physical form, such as a granule, that retains the microorganism, carbon source, and rock phosphate ore, but permits water and soluble phosphate to diffuse into the soil. A method of using the composition for providing phosphate fertilizer to plants is also disclosed.

  1. Role of Phosphate Transport System Component PstB1 in Phosphate Internalization by Nostoc punctiforme

    PubMed Central

    Hudek, L.; Premachandra, D.; Webster, W. A. J.

    2016-01-01

    ABSTRACT In bacteria, limited phosphate availability promotes the synthesis of active uptake systems, such as the Pst phosphate transport system. To understand the mechanisms that facilitate phosphate accumulation in the cyanobacterium Nostoc punctiforme, phosphate transport systems were identified, revealing a redundancy of Pst phosphate uptake systems that exists across three distinct operons. Four separate PstB system components were identified. pstB1 was determined to be a suitable target for creating phenotypic mutations that could result in the accumulation of excessive levels of phosphate through its overexpression or in a reduction of the capacity to accumulate phosphate through its deletion. Using quantitative real-time PCR (qPCR), it was determined that pstB1 mRNA levels increased significantly over 64 h in cells cultured in 0 mM added phosphate and decreased significantly in cells exposed to high (12.8 mM) phosphate concentrations compared to the level in cells cultured under normal (0.8 mM) conditions. Possible compensation for the loss of PstB1 was observed when pstB2, pstB3, and pstB4 mRNA levels increased, particularly in cells starved of phosphate. The overexpression of pstB1 increased phosphate uptake by N. punctiforme and was shown to functionally complement the loss of PstB in E. coli PstB knockout (PstB−) mutants. The knockout of pstB1 in N. punctiforme did not have a significant effect on cellular phosphate accumulation or growth for the most part, which is attributed to the compensation for the loss of PstB1 by alterations in the pstB2, pstB3, and pstB4 mRNA levels. This study provides novel in vivo evidence that PstB1 plays a functional role in phosphate uptake in N. punctiforme. IMPORTANCE Cyanobacteria have been evolving over 3.5 billion years and have become highly adept at growing under limiting nutrient levels. Phosphate is crucial for the survival and prosperity of all organisms. In bacteria, limited phosphate availability promotes

  2. 21 CFR 184.1434 - Magnesium phosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Magnesium phosphate. 184.1434 Section 184.1434... Listing of Specific Substances Affirmed as GRAS § 184.1434 Magnesium phosphate. (a) Magnesium phosphate includes both magnesium phosphate, dibasic, and magnesium phosphate, tribasic. Magnesium phosphate, dibasic...

  3. 21 CFR 184.1434 - Magnesium phosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Magnesium phosphate. 184.1434 Section 184.1434... Listing of Specific Substances Affirmed as GRAS § 184.1434 Magnesium phosphate. (a) Magnesium phosphate includes both magnesium phosphate, dibasic, and magnesium phosphate, tribasic. Magnesium phosphate, dibasic...

  4. 21 CFR 184.1434 - Magnesium phosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Magnesium phosphate. 184.1434 Section 184.1434... Listing of Specific Substances Affirmed as GRAS § 184.1434 Magnesium phosphate. (a) Magnesium phosphate includes both magnesium phosphate, dibasic, and magnesium phosphate, tribasic. Magnesium phosphate, dibasic...

  5. Immediate and delayed signal of slab breakoff in Oligo/Miocene Molasse deposits from the European Alps

    PubMed Central

    Schlunegger, Fritz; Castelltort, Sébastien

    2016-01-01

    High-resolution 32–20 Ma-old stratigraphic records from the Molasse foreland basin situated north of the Alps, and Gonfolite Lombarda conglomerates deposited on the southern Alpine margin, document two consecutive sedimentary responses - an immediate and delayed response - to slab breakoff beneath the central Alps c. 32–30 Ma ago. The first signal, which occurred due to rebound and surface uplift in the Alps, was a regional and simultaneous switch from basin underfill to overfill at 30 Ma paired with shifts to coarse-grained depositional environments in the foreland basin. The second signal, however, arrived several million years after slab breakoff and was marked by larger contributions of crystalline clasts in the conglomerates, larger clast sizes, larger sediment fluxes and shifts to more proximal facies. We propose that this secondary pulse reflects a delayed whiplash-type erosional response to surface uplift, where erosion and sediment flux became amplified through positive feedbacks once larger erosional thresholds of crystalline bedrock were exceeded. PMID:27510939

  6. Alkaline polymer electrolyte membranes for fuel cell applications.

    PubMed

    Wang, Yan-Jie; Qiao, Jinli; Baker, Ryan; Zhang, Jiujun

    2013-07-07

    In this review, we examine the most recent progress and research trends in the area of alkaline polymer electrolyte membrane (PEM) development in terms of material selection, synthesis, characterization, and theoretical approach, as well as their fabrication into alkaline PEM-based membrane electrode assemblies (MEAs) and the corresponding performance/durability in alkaline polymer electrolyte membrane fuel cells (PEMFCs). Respective advantages and challenges are also reviewed. To overcome challenges hindering alkaline PEM technology advancement and commercialization, several research directions are then proposed.

  7. Environmental benefits of using magnesium carbonate minerals as new wildfire retardants instead of commercially available, phosphate-based compounds.

    PubMed

    Liodakis, S; Tsoukala, M

    2010-10-01

    A serial batch leaching experiment has been carried out to evaluate the release of elements from the ash of Pinus halepensis needles burned under two test conditions-with and without treatment of the forest species with the carbonate minerals (huntite and hydromagnesite) in aqueous solution (pH 6). The ash (before and after leaching) and leachates were analyzed using atomic absorption spectroscopy and X-ray diffraction. Compared with data from samples treated with the commercially available, phosphate-based fire retardant diammonium phosphate (DAP), we found that use of huntite or hydromagnesite was much more successful in obstructing the release of the toxic elements present in the ash, probably because of the alkaline conditions resulting from decomposition of the minerals during burning. In contrast, DAP tended to be more able to facilitate the extraction of some toxic metals (e.g., Zn, Cu, Mn), probably because of the acidic conditions resulting from its decomposition to phosphoric acid. Data from this study thus lend strong support to the use of magnesium carbonate minerals as new wildfire retardants, because they were shown to be more friendly to the environment (e.g., soil, ground, and underground water streams) than those currently in use (e.g., phosphate or sulfate salt type).

  8. pH-Dependent Binding of Chloride to a Marine Alkaline Phosphatase Affects the Catalysis, Active Site Stability, and Dimer Equilibrium.

    PubMed

    Hjörleifsson, Jens G; Ásgeirsson, Bjarni

    2017-09-26

    The effect of ionic strength on enzyme activity and stability varies considerably between enzymes. Ionic strength is known to affect the catalytic activity of some alkaline phosphatases (APs), such as Escherichia coli AP, but how ions affect APs is debated. Here, we studied the effect of various ions on a cold-adapted AP from Vibrio splendidus (VAP). Previously, we have found that the active form of VAP is extremely unstable at low ionic strengths. Here we show that NaCl increased the activity and stability of VAP and that the effect was pH-dependent in the range of pH 7-10. The activity profile as a function of pH formed two maxima, indicating a possible conformational change. Bringing the pH from the neutral to the alkaline range was accompanied by a large increase in both the K i for inorganic phosphate (product inhibition) and the K M for p-nitrophenyl phosphate. The activity transitions observed as the pH was varied correlated with structural changes as monitored by tryptophan fluorescence. Thermal and urea-induced inactivation was shown to be accompanied by neither dissociation of the active site metal ions nor dimer dissociation. This would suggest that the inactivation involved subtle changes in active site conformation. Furthermore, the VAP dimer equilibrium was studied for the first time and shown to highly favor dimerization, which was dependent on pH and NaCl concentration. Taken together, the data support a model in which anions bind to some specific acceptor in the active site of VAP, resulting in great stabilization and catalytic rate enhancement, presumably through a different mechanism.

  9. How do arbuscular mycorrhizal fungi handle phosphate? New insight into fine-tuning of phosphate metabolism.

    PubMed

    Ezawa, Tatsuhiro; Saito, Katsuharu

    2018-04-27

    Contents Summary I. Introduction II. Foraging for phosphate III. Fine-tuning of phosphate homeostasis IV. The frontiers: phosphate translocation and export V. Conclusions and outlook Acknowledgements References SUMMARY: Arbuscular mycorrhizal fungi form symbiotic associations with most land plants and deliver mineral nutrients, in particular phosphate, to the host. Therefore, understanding the mechanisms of phosphate acquisition and delivery in the fungi is critical for full appreciation of the mutualism in this association. Here, we provide updates on physical, chemical, and biological strategies of the fungi for phosphate acquisition, including interactions with phosphate-solubilizing bacteria, and those on the regulatory mechanisms of phosphate homeostasis based on resurveys of published genome sequences and a transcriptome with reference to the latest findings in a model fungus. For the mechanisms underlying phosphate translocation and export to the host, which are major research frontiers in this field, not only recent advances but also testable hypotheses are proposed. Lastly, we briefly discuss applicability of the latest tools to gene silencing in the fungi, which will be breakthrough techniques for comprehensive understanding of the molecular basis of fungal phosphate metabolism. © 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

  10. 21 CFR 184.1434 - Magnesium phosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Magnesium phosphate. 184.1434 Section 184.1434 Food... Specific Substances Affirmed as GRAS § 184.1434 Magnesium phosphate. (a) Magnesium phosphate includes both magnesium phosphate, dibasic, and magnesium phosphate, tribasic. Magnesium phosphate, dibasic (MgHPO4·3H2O...

  11. 21 CFR 184.1434 - Magnesium phosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Magnesium phosphate. 184.1434 Section 184.1434... GRAS § 184.1434 Magnesium phosphate. (a) Magnesium phosphate includes both magnesium phosphate, dibasic, and magnesium phosphate, tribasic. Magnesium phosphate, dibasic (MgHPO4·3H2O, CAS Reg. No. 7782-0975...

  12. Phosphated Cellulose as an Efficient Biomaterial for Aqueous Drug Ranitidine Removal

    PubMed Central

    Bezerra, Roosevelt D. S.; Silva, Márcia M. F.; Morais, Alan I. S.; Osajima, Josy A.; Santos, Maria R. M. C.; Airoldi, Claudio; Silva Filho, Edson C.

    2014-01-01

    Crystalline cellulose chemically modified through a reaction with sodium trimetaphosphate (STMP) in an acidic or basic condition yielded Cel-P4 and Cel-P10. These phosphated solids were characterized by elemental analysis, X-ray diffraction (XRD), infrared (IR) spectroscopy, scanning electron microscopy (SEM), nuclear magnetic resonance (NMR) at the solid state for phosphorus nucleus and dispersive X-ray energy. The elemental results demonstrated that the phosphorylation reaction was more efficient in the basic medium, as supported by the amount of phosphorous content. The synthesized biomaterials decreased in crystallinity in comparison to the precursor cellulose, with an increase in roughness and present two distinct phosphorus environments in the formed structure. The phosphated cellulose in an alkaline condition was applied to sorb the drug ranitidine. This process was applied in varying pH, time, temperature and concentration. The best sorption kinetic model to fit the experimental data was the pseudo-second-order with a coefficient correlation of 0.8976, and the Langmuir isotherm model was the most adjusted to the variation in concentration. The efficient drug sorption has a low dependence on temperature, with maximum values of 85.0, 82.0 mg and 85.7 mg·g−1 for Cel-P10 at 298, 308 and 318 K, respectively. The best sorption occurred at pH = 6 with a saturation time of 210 min. PMID:28788283

  13. Role of Phosphate Transport System Component PstB1 in Phosphate Internalization by Nostoc punctiforme.

    PubMed

    Hudek, L; Premachandra, D; Webster, W A J; Bräu, L

    2016-11-01

    In bacteria, limited phosphate availability promotes the synthesis of active uptake systems, such as the Pst phosphate transport system. To understand the mechanisms that facilitate phosphate accumulation in the cyanobacterium Nostoc punctiforme, phosphate transport systems were identified, revealing a redundancy of Pst phosphate uptake systems that exists across three distinct operons. Four separate PstB system components were identified. pstB1 was determined to be a suitable target for creating phenotypic mutations that could result in the accumulation of excessive levels of phosphate through its overexpression or in a reduction of the capacity to accumulate phosphate through its deletion. Using quantitative real-time PCR (qPCR), it was determined that pstB1 mRNA levels increased significantly over 64 h in cells cultured in 0 mM added phosphate and decreased significantly in cells exposed to high (12.8 mM) phosphate concentrations compared to the level in cells cultured under normal (0.8 mM) conditions. Possible compensation for the loss of PstB1 was observed when pstB2, pstB3, and pstB4 mRNA levels increased, particularly in cells starved of phosphate. The overexpression of pstB1 increased phosphate uptake by N. punctiforme and was shown to functionally complement the loss of PstB in E. coli PstB knockout (PstB - ) mutants. The knockout of pstB1 in N. punctiforme did not have a significant effect on cellular phosphate accumulation or growth for the most part, which is attributed to the compensation for the loss of PstB1 by alterations in the pstB2, pstB3, and pstB4 mRNA levels. This study provides novel in vivo evidence that PstB1 plays a functional role in phosphate uptake in N. punctiforme IMPORTANCE: Cyanobacteria have been evolving over 3.5 billion years and have become highly adept at growing under limiting nutrient levels. Phosphate is crucial for the survival and prosperity of all organisms. In bacteria, limited phosphate availability promotes the

  14. Antiresorption implant coatings based on calcium alendronate and octacalcium phosphate deposited by matrix assisted pulsed laser evaporation.

    PubMed

    Boanini, Elisa; Torricelli, Paola; Forte, Lucia; Pagani, Stefania; Mihailescu, Natalia; Ristoscu, Carmen; Mihailescu, Ion N; Bigi, Adriana

    2015-12-01

    The integration of an implant material with bone tissue depends on the chemistry and physics of the implant surface. In this study we applied matrix assisted pulsed laser evaporation (MAPLE) in order to synthesize calcium alendronate monohydrate (a bisphosphonate obtained by calcium sequestration from octacalcium phosphate by alendronate) and calcium alendronate monohydrate/octacalcium phosphate composite thin films on titanium substrates. Octacalcium phosphate coatings were prepared as reference material. The powders, which were synthesized in aqueous medium, were suspended in deionised water, frozen at liquid nitrogen temperature and used as targets for MAPLE experiments. The transfer was conducted with a KrF* excimer laser source (λ = 248 nm, τFWHM ≤ 25 ns) in mild conditions of temperature and pressure. XRD, FTIR and SEM analyses confirmed that the coatings contain the same crystalline phases as the as-prepared powder samples. Osteoblast derived from stem cells and osteoclast derived from monocytes of osteoporotic subjects were co-cultured on the coatings up to 14 days. Osteoclast displayed significantly reduced proliferation and differentiation in the presence of calcium alendronate monohydrate, pointing to a clear role of the coatings containing this bisphosphonate on inhibiting excessive bone resorption. At variance, osteoblast production of alkaline phosphatase and type I pro-collagen were promoted by the presence of bisphosphonate, which also decreased the production of interleukin 6. The positive influence towards osteoblast differentiation was even more enhanced in the composite coatings, thanks to the presence of octacalcium phosphate. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Fission track ages and Exhumation mechanisms of the Tauern Window, Eastern Alps

    NASA Astrophysics Data System (ADS)

    Bertrand, Audrey; Rosenberg, Claudio; Garcia, Sebastian

    2010-05-01

    The Tauern Window (TW) is a thermal and structural dome which exposes Penninic basement, its cover units as well as parts of the overlying Austroalpine basement in the central part of the Eastern Alps. The peak of metamorphism was attained approximately at 30Ma (Selverstone et .al, 1992), followed by cooling and exhumation throughout Miocene time. Most of the tertiary exhumation of the Eastern Alps was localized in the TW, from Early Oligocene to late Miocene time. A current debate centers on the exhumation mechanisms of Penninic rocks in the core of the TW, namely to assess whether orogen-parallel extension (e.g., Selverstone, 1988) or a combination of folding and erosion (eg., Rosenberg et al., 2004) with subordinate extension were the controlling processes. E-W extension is well documented at the western (Brenner Fault) and eastern (Katschberg Fault) margins of the window (e.g., Behrmann, 1988; Selverstone, 1988; Genser and Neubauer, 1989). In contrast, upright folding dominates the internal structure of the dome, and in particular along its western part, where fold amplitudes, mostly eroded during folding, attained up to 10 km. This study attempts to assess the relative importance of folding and erosion and of orogen-parallel extension during exhumation by analyzing the spatial and temporal cooling patterns of apatite and zircon fission track ages. The compilation of published apatite and zircon fission track ages indicates a concentric younging of both the apatite and zircon ages toward the core of the TW. The concentric isochrones follow the map trace of the axial planes of the upright folds of the western and eastern TW. This cooling pattern is in contrast to the one expected by a process of extensional unroofing, which in map view would results in isochrons parallel to the extensional faults and progressively younging towards them (e.g., Foster et al., 2001). We therefore propose that folding and erosion were primarily responsible for exhuming the Penninic

  16. Net alkalinity and net acidity 1: Theoretical considerations

    USGS Publications Warehouse

    Kirby, C.S.; Cravotta, C.A.

    2005-01-01

    Net acidity and net alkalinity are widely used, poorly defined, and commonly misunderstood parameters for the characterization of mine drainage. The authors explain theoretical expressions of 3 types of alkalinity (caustic, phenolphthalein, and total) and acidity (mineral, CO2, and total). Except for rarely-invoked negative alkalinity, theoretically defined total alkalinity is closely analogous to measured alkalinity and presents few practical interpretation problems. Theoretically defined "CO 2-acidity" is closely related to most standard titration methods with an endpoint pH of 8.3 used for determining acidity in mine drainage, but it is unfortunately named because CO2 is intentionally driven off during titration of mine-drainage samples. Using the proton condition/mass- action approach and employing graphs to illustrate speciation with changes in pH, the authors explore the concept of principal components and how to assign acidity contributions to aqueous species commonly present in mine drainage. Acidity is defined in mine drainage based on aqueous speciation at the sample pH and on the capacity of these species to undergo hydrolysis to pH 8.3. Application of this definition shows that the computed acidity in mg L -1 as CaCO3 (based on pH and analytical concentrations of dissolved FeII, FeIII, Mn, and Al in mg L -1):aciditycalculated=50{1000(10-pH)+[2(FeII)+3(FeIII)]/56+2(Mn)/ 55+3(Al)/27}underestimates contributions from HSO4- and H+, but overestimates the acidity due to Fe3+ and Al3+. However, these errors tend to approximately cancel each other. It is demonstrated that "net alkalinity" is a valid mathematical construction based on theoretical definitions of alkalinity and acidity. Further, it is shown that, for most mine-drainage solutions, a useful net alkalinity value can be derived from: (1) alkalinity and acidity values based on aqueous speciation, (2) measured alkalinity minus calculated acidity, or (3) taking the negative of the value obtained in a

  17. Bacterial Composition and Survival on Sahara Dust Particles Transported to the European Alps

    PubMed Central

    Meola, Marco; Lazzaro, Anna; Zeyer, Josef

    2015-01-01

    Deposition of Sahara dust (SD) particles is a frequent phenomenon in Europe, but little is known about the viability and composition of the bacterial community transported with SD. The goal of this study was to characterize SD-associated bacteria transported to the European Alps, deposited and entrapped in snow. During two distinct events in February and May 2014, SD particles were deposited and promptly covered by falling snow, thus preserving them in distinct ochre layers within the snowpack. In June 2014, we collected samples at different depths from a snow profile at the Jungfraujoch (Swiss Alps; 3621 m a.s.l.). After filtration, we performed various microbiological and physicochemical analyses of the snow and dust particles therein that originated in Algeria. Our results show that bacteria survive and are metabolically active after the transport to the European Alps. Using high throughput sequencing, we observed distinct differences in bacterial community composition and structure in SD-layers as compared to clean snow layers. Sporulating bacteria were not enriched in the SD-layers; however, phyla with low abundance such as Gemmatimonadetes and Deinococcus-Thermus appeared to be specific bio-indicators for SD. Since many members of these phyla are known to be adapted to arid oligotrophic environments and UV radiation, they are well suited to survive the harsh conditions of long-range airborne transport. PMID:26733988

  18. Acetyl Phosphate as a Primordial Energy Currency at the Origin of Life

    NASA Astrophysics Data System (ADS)

    Whicher, Alexandra; Camprubi, Eloi; Pinna, Silvana; Herschy, Barry; Lane, Nick

    2018-03-01

    Metabolism is primed through the formation of thioesters via acetyl CoA and the phosphorylation of substrates by ATP. Prebiotic equivalents such as methyl thioacetate and acetyl phosphate have been proposed to catalyse analogous reactions at the origin of life, but their propensity to hydrolyse challenges this view. Here we show that acetyl phosphate (AcP) can be synthesised in water within minutes from thioacetate (but not methyl thioacetate) under ambient conditions. AcP is stable over hours, depending on temperature, pH and cation content, giving it an ideal poise between stability and reactivity. We show that AcP can phosphorylate nucleotide precursors such as ribose to ribose-5-phosphate and adenosine to adenosine monophosphate, at modest ( 2%) yield in water, and at a range of pH. AcP can also phosphorylate ADP to ATP in water over several hours at 50 °C. But AcP did not promote polymerization of either glycine or AMP. The amino group of glycine was preferentially acetylated by AcP, especially at alkaline pH, hindering the formation of polypeptides. AMP formed small stacks of up to 7 monomers, but these did not polymerise in the presence of AcP in aqueous solution. We conclude that AcP can phosphorylate biologically meaningful substrates in a manner analogous to ATP, promoting the origins of metabolism, but is unlikely to have driven polymerization of macromolecules such as polypeptides or RNA in free solution. This is consistent with the idea that a period of monomer (cofactor) catalysis preceded the emergence of polymeric enzymes or ribozymes at the origin of life.

  19. Ice cap melting and low-viscosity crustal root explain the narrow geodetic uplift of the Western Alps

    NASA Astrophysics Data System (ADS)

    Chéry, J.; Genti, M.; Vernant, P.

    2016-04-01

    More than 10 years of geodetic measurements demonstrate an uplift rate of 1-3 mm/yr of the high topography region of the Western Alps. By contrast, no significant horizontal motion has been detected. Two uplift mechanisms have been proposed: (1) the isostatic response to denudation responsible for only a fraction of the observed uplift and (2) the rebound induced by the Wurmian ice cap melting which predicts a broader uplifting region than the one evidenced by geodetic observations. Using a numerical model to fit the geodetic data, we show that a crustal viscosity contrast between the foreland and the central part of the Alps, the latter being weaker with a viscosity of 1021 Pa s, is needed. The vertical rates are enhanced if the strong uppermost mantle beneath the Moho is interrupted across the Alps, therefore allowing a weak vertical rheological anomaly over the entire lithosphere.

  20. Francisella DnaK Inhibits Tissue-nonspecific Alkaline Phosphatase*

    PubMed Central

    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