Science.gov

Sample records for alkaline phosphate alp

  1. Alkaline phosphatase revisited: hydrolysis of alkyl phosphates.

    PubMed

    O'Brien, Patrick J; Herschlag, Daniel

    2002-03-05

    Escherichia coli alkaline phosphatase (AP) is the prototypical two metal ion catalyst with two divalent zinc ions bound approximately 4 A apart in the active site. Studies spanning half a century have elucidated many structural and mechanistic features of this enzyme, rendering it an attractive model for investigating the potent catalytic power of bimetallic centers. Unfortunately, fundamental mechanistic features have been obscured by limitations with the standard assays. These assays generate concentrations of inorganic phosphate (P(i)) in excess of its inhibition constant (K(i) approximately 1 muM). This tight binding by P(i) has affected the majority of published kinetic constants. Furthermore, binding limits k(cat)/K(m) for reaction of p-nitrophenyl phosphate, the most commonly employed substrate. We describe a sensitive (32)P-based assay for hydrolysis of alkyl phosphates that avoids the complication of product inhibition. We have revisited basic mechanistic features of AP with these alkyl phosphate substrates. The results suggest that the chemical step for phosphorylation of the enzyme limits k(cat)/K(m). The pH-rate profile and additional results suggest that the serine nucleophile is active in its anionic form and has a pK(a) of < or = 5.5 in the free enzyme. An inactivating pK(a) of 8.0 is observed for binding of both substrates and inhibitors, and we suggest that this corresponds to ionization of a zinc-coordinated water molecule. Counter to previous suggestions, inorganic phosphate dianion appears to bind to the highly charged AP active site at least as strongly as the trianion. The dependence of k(cat)/K(m) on the pK(a) of the leaving group follows a Brønsted correlation with a slope of beta(lg) = -0.85 +/- 0.1, differing substantially from the previously reported value of -0.2 obtained from data with a less sensitive assay. This steep leaving group dependence is consistent with a largely dissociative transition state for AP-catalyzed hydrolysis of

  2. A sensitive enzyme-catalytic nanogold-resonance scattering spectral assay for alkaline phosphate.

    PubMed

    Jiang, Zhiliang; Wu, Meng; Liu, Gaosan; Liang, Aihui

    2012-06-01

    In pH 8.9 Tris-HCl buffer solutions, alkaline phosphatase (ALP) catalyzed the hydrolysis of ascorbic acid 2-phosphate (AAP) substrate to form ascorbic acid. Then H(3)PO(4) was added to stop the enzymatic reaction and HAuCl(4) was used to react with ascorbic acid to generate gold nanoparticles that exhibited a resonance scattering (RS) peak at 600 nm. Under the selected conditions, when the activity of ALP increased, the formed ascorbic acid and gold nanoparticles also increased. Thus, the RS intensity at 600 nm enhanced linearly. The linear range was 0.06-22 U/L, with a detection limit of 0.03 U/L. The ALP in serum was analyzed, and the results were in agreement with those of the fluorescence method.

  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. 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...

  5. 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...

  6. Cloning & Characterization of the Cry1Ac-binding Alkaline Phosphatase (HvALP) from Heliothis virescens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Membrane bound alkaline phosphatases (mALPs) in the insect midgut have been reported as functional receptors for Cry toxins from the bacterium Bacillus thuringiensis. We previously reported the identification of HvALP in the midgut of Heliothis virescens larvae as a Cry1Ac binding protein that is d...

  7. 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.

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

    PubMed

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

    2016-11-15

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

  9. 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.

  10. Alkaline Phosphatase Revisited:  Hydrolysis of Alkyl Phosphates (†).

    PubMed

    O'Brien, Patrick J; Herschlag, Daniel

    2002-03-05

    Escherichia coli alkaline phosphatase (AP) is the prototypical two metal ion catalyst with two divalent zinc ions bound ∼4 Å apart in the active site. Studies spanning half a century have elucidated many structural and mechanistic features of this enzyme, rendering it an attractive model for investigating the potent catalytic power of bimetallic centers. Unfortunately, fundamental mechanistic features have been obscured by limitations with the standard assays. These assays generate concentrations of inorganic phosphate (Pi) in excess of its inhibition constant (K i ≈ 1 μM). This tight binding by Pi has affected the majority of published kinetic constants. Furthermore, binding limits k cat/K m for reaction of p-nitrophenyl phosphate, the most commonly employed substrate. We describe a sensitive (32)P-based assay for hydrolysis of alkyl phosphates that avoids the complication of product inhibition. We have revisited basic mechanistic features of AP with these alkyl phosphate substrates. The results suggest that the chemical step for phosphorylation of the enzyme limits k cat/K m. The pH-rate profile and additional results suggest that the serine nucleophile is active in its anionic form and has a pK a of ≤5.5 in the free enzyme. An inactivating pK a of 8.0 is observed for binding of both substrates and inhibitors, and we suggest that this corresponds to ionization of a zinc-coordinated water molecule. Counter to previous suggestions, inorganic phosphate dianion appears to bind to the highly charged AP active site at least as strongly as the trianion. The dependence of k cat/K m on the pK a of the leaving group follows a Brønsted correlation with a slope of βlg = -0.85 ± 0.1, differing substantially from the previously reported value of -0.2 obtained from data with a less sensitive assay. This steep leaving group dependence is consistent with a largely dissociative transition state for AP-catalyzed hydrolysis of phosphate monoesters. The new (32)P

  11. L-Phenylalanine inhibition of human alkaline phosphatases with p-nitrophenyl phosphate as substrate.

    PubMed

    Komoda, T; Hokari, S; Sonoda, M; Sakagishi, Y; Tamura, T

    1982-12-01

    With p-nitrophenyl phosphate as the substrate, there reportedly is no organ-specific inhibition of alkaline phosphatase (EC 3.1.3.1) activity by L-phenylalanine. However, we found that at pH 10.0, with p-nitrophenyl phosphate as the substrate, L-phenylalanine obviously inhibits the alkaline phosphatase isoenzyme from human placenta, whereas there is little if any inhibition of the isoenzyme from human intestine. Because of the differing effects of substrates (p-nitrophenyl phosphate and phenyl phosphate) and their enzymic products (p-nitrophenol and phenol) for L-phenylalanine action on the placental alkaline phosphatase isoenzyme, we suggest that the isoenzyme--inhibitor--substrate complex and the effect of released phosphate on L-phenylalanine inhibition of the isoenzyme activity differ from each other.

  12. Spatial Patterns of Alkaline Phosphatase Expression within Bacterial Colonies and Biofilms in Response to Phosphate Starvation

    PubMed Central

    Huang, Ching-Tsan; Xu, Karen D.; McFeters, Gordon A.; Stewart, Philip S.

    1998-01-01

    The expression of alkaline phosphatase in response to phosphate starvation was shown to be spatially and temporally heterogeneous in bacterial biofilms and colonies. A commercial alkaline phosphatase substrate that generates a fluorescent, insoluble product was used in conjunction with frozen sectioning techniques to visualize spatial patterns of enzyme expression in both Klebsiella pneumoniae and Pseudomonas aeruginosa biofilms. Some of the expression patterns observed revealed alkaline phosphatase activity at the boundary of the biofilm opposite the place where the staining substrate was delivered, indicating that the enzyme substrate penetrated the biofilm fully. Alkaline phosphatase accumulated linearly with time in K. pneumoniae colonies transferred from high-phosphate medium to low-phosphate medium up to specific activities of 50 μmol per min per mg of protein after 24 h. In K. pneumoniae biofilms and colonies, alkaline phosphatase was initially expressed in the region of the biofilm immediately adjacent to the carbon and energy source (glucose). In time, the region of alkaline phosphatase expression expanded inward until it spanned most, but not all, of the biofilm or colony depth. In contrast, expression of alkaline phosphatase in P. aeruginosa biofilms occurred in a thin, sharply delineated band at the biofilm-bulk fluid interface. In this case, the band of activity never occupied more than approximately one-sixth of the biofilm. These results are consistent with the working hypothesis that alkaline phosphatase expression patterns are primarily controlled by the local availability of either the carbon and energy source or the electron acceptor. PMID:9546188

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

    PubMed

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

    2015-10-01

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

  14. 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.

  15. Phosphate monoester hydrolysis by trinuclear alkaline phosphatase; DFT study of transition States and reaction mechanism.

    PubMed

    Chen, Shi-Lu; Liao, Rong-Zhen

    2014-08-04

    Alkaline phosphatase (AP) is a trinuclear metalloenzyme that catalyzes the hydrolysis of a broad range of phosphate monoesters to form inorganic phosphate and alcohol (or phenol). In this paper, by using density functional theory with a model based on a crystal structure, the AP-catalyzed hydrolysis of phosphate monoesters is investigated by calculating two substrates, that is, methyl and p-nitrophenyl phosphates, which represent alkyl and aryl phosphates, respectively. The calculations confirm that the AP reaction employs a "ping-pong" mechanism involving two chemical displacement steps, that is, the displacement of the substrate leaving group by a Ser102 alkoxide and the hydrolysis of the phosphoseryl intermediate by a Zn2-bound hydroxide. Both displacement steps proceed via a concerted associative pathway no matter which substrate is used. Other mechanistic aspects are also studied. Comparison of our calculations with linear free energy relationships experiments shows good agreement.

  16. High-Strength / High Alkaline Resistant Fe-Phosphate Glass Fibers as Concrete Reinforcement

    SciTech Connect

    Mariano Velez

    2008-03-31

    Calcium-iron-phosphate glasses were developed whose chemical durabilities in alkaline solutions (pH 13) were comparable or superior to those of commercial alkaline-resistant (AR) silica-based glasses. However, the tensile strength of Ca-Fe-phosphate fibers, after being exposed to alkaline environments, including wet Portland cement pastes, is lower than that of current AR silicate fibers. Another series of Ca-Fe-phosphate glasses were developed with excellent chemical durability in strong acidic solutions (H2SO4, HF), indicating potential applications where silica-based fibers degrade very quickly, including E-glass. The new Ca-Fe-phosphate glasses can be melted and processed 300 to 500°C lower than silica-based glasses. This offers the possibility of manufacturing glass fibers with lower energy costs by 40-60% and the potential to reduce manufacturing waste and lower gas emissions. It was found that Ca-Fe-phosphate melts can be continuously pulled into fibers depending on the slope of the viscosity-temperature curve and with viscosity ~100 poise, using multi-hole Pt/Rh bushings.

  17. Supermolecule density functional calculations suggest a key role for solvent in alkaline hydrolysis of p-nitrophenyl phosphate.

    PubMed

    Zhang, Lidong; Xie, Daiqian; Xu, Dingguo; Guo, Hua

    2007-04-28

    Supermolecule density functional theory calculations show that solvent is responsible for the concerted transition state in alkaline hydrolysis of p-nitrophenyl phosphate suggested by heavy atom kinetic isotope effects.

  18. ALP (Alkaline Phosphatase) Test

    MedlinePlus

    PLEASE NOTE: Your web browser does not have JavaScript enabled. Unless you enable Javascript , your ability to navigate and access the features of this website will be limited. ... Proceeds from website advertising help sustain Lab Tests Online. AACC is a not-for-profit organization and does not endorse non-AACC products and services. Advertising & Sponsorship: ...

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

    PubMed

    Upadhyay, Lata Sheo Bachan; Verma, Nishant

    2015-06-15

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

  20. Enzymatic kinetic parameters for polyfluorinated alkyl phosphate hydrolysis by alkaline phosphatase.

    PubMed

    Jackson, Derek A; Mabury, Scott A

    2012-09-01

    The hydrolysis kinetics of three polyfluorinated alkyl phosphate monoesters (monoPAPs), differing in fluorinated chain length, were measured using bovine intestinal alkaline phosphatase to catalyze the reaction. Kinetic values were also measured for analogous hydrogenated phosphate monoesters to elucidate the effects of the fluorinated chain on the rate of enzymatic hydrolysis. Michaelis constants (K(m)) were obtained by a competition kinetics technique in the presence of p-nitrophenyl phosphate (PNPP) using UV-vis spectroscopy. Compared with K(m) (PNPP), Michaelis constants for monoPAPs ranged from 0.9 to 2.1 compared with hydrogenated phosphates, which ranged from 4.0 to 13.0. Apparent bimolecular rate constants (k(cat)/K(m)) were determined by monitoring rates of product alcohol formation at low substrate concentrations using gas chromatography-mass spectrometry. The experimental values for k(cat)/K(m) averaged as 1.1 × 10(7) M(-1) s(-1) for monoPAPs compared with 3.8 × 10(5) M(-1) s(-1) for hexyl phosphate. This suggests that the electron-withdrawing nature of the fluorinated chain enhanced the alcohol leaving group ability. The results were used in a simple model to suggest that monoPAPs in a typical mammalian digestive tract would hydrolyze in approximately 100 s, supporting a previous study that showed its absence after a dosing study in rats.

  1. Acid inactivation of and incorporation of phosphate into alkaline phosphatase from Escherichia coli

    PubMed Central

    Pigretti, M. M.; Milstein, C.

    1965-01-01

    1. Alkaline phosphatase of Escherichia coli undergoes below pH 6·0 a reversible acid inactivation that has been studied and related to the extent of uptake of inorganic phosphate occurring below pH 6·0. 2. The rate of inactivation is rapid in the first few minutes but later it decreases markedly. Temperature, pH, composition of buffer and other factors have an important effect on the inactivation. 3. About 60% of the activity lost at pH values above 3·5 is rapidly recovered when the enzyme is taken back to pH 8·0, independently (within certain limits) of the extent of the inactivation. 4. Phosphate and Zn2+, although very good protectors of the inactivation by acid, are not by themselves able to reverse the acid inactivation. 5. Inorganic phosphate seems not to be incorporated into the acid-inactivated enzyme. 6. Incorporation of more than one mole of phosphate/mole of enzyme has been obtained, but the phosphate residues seem to be incorporated to serine residues with a common sequence, suggesting two identical active serine residues/molecule of active enzyme. ImagesFig. 3.Fig. 4. PMID:14342215

  2. Recovery of phosphorus and nitrogen from alkaline hydrolysis supernatant of excess sludge by magnesium ammonium phosphate.

    PubMed

    Bi, Wei; Li, Yiyong; Hu, Yongyou

    2014-08-01

    Magnesium ammonium phosphate (MAP) method was used to recover orthophosphate (PO₄(3-)-P) and ammonium nitrogen (NH4(+)-N) from the alkaline hydrolysis supernatant of excess sludge. To reduce alkali consumption and decrease the pH of the supernatant, two-stage alkaline hydrolysis process (TSAHP) was designed. The results showed that the release efficiencies of PO₄(3-)-P and NH₄(+)-N were 41.96% and 7.78%, respectively, and the pH of the supernatant was below 10.5 under the running conditions with initial pH of 13, volume ratio (sludge dosage/water dosage) of 1.75 in second-stage alkaline hydrolysis reactor, 20 g/L of sludge concentration in first-stage alkaline hydrolysis reactor. The order of parameters influencing MAP reaction was analyzed and the optimized conditions of MAP reaction were predicted through the response surface methodology. The recovery rates of PO₄(3-)-P and NH₄(+)-N were 46.88% and 16.54%, respectively under the optimized conditions of Mg/P of 1.8, pH 9.7 and reaction time of 15 min.

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

    PubMed

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

    1977-09-01

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

  4. [A study on the appropriate fixation for the procedures for the better preservation of cellular antigenicity and morphology of the blood smear in immunocytochemistry: an improvement of the immunostain technique using alkaline-phosphatase (ALP) as a labeling enzyme].

    PubMed

    Aoki, J; Sasaki, N; Hino, N; Nanba, K

    1991-01-01

    The authors previously reported a new coloration method which utilized hexazotized newfuchsin as a coupler for the immuno-enzyme-cytochemistry. This procedure used alkaline-phosphatase (ALP) as the labeling enzyme. The insolubility of the reaction product to organic solvents made it possible to prepare permanent slides. However, this suffered from several drawbacks, due to the fixation procedures, in the preservation of better morphology and antigenicity of the cell. The present study was undertaken to overcome such problems by modifying the fixation procedure. The study utilized twenty monoclonal and polyclonal antibodies commonly used in immunohematological staining. Various fixative solutions and timing of fixation were evaluated. The results indicated that; 1) the best fixative solution was a mixture of buffered paraformaldehyde (PFA) and acetone (10 ml 40% PFA solution, 10 ml pH 6.6 0.02 M phosphate buffer, 20 ml distilled water, 60 ml acetone, with pH adjusted to 6.6-7.4 with HCl) and 2) the fixation should be performed just before the immunostain. The results further showed that unstained smear slides, when freshly air dried and stored in a desicator, could maintain various differentiation antigens (CD2, 3, 4, 5, 8, 10, 14, 15, 16, 19, 25, L26, HLA-DR) for at least 4 weeks without any change in the immuno-reactivity. Thus, we conclude that this improved fixation procedure is an optimum fixative and should be used in routine application of the immunostain method for blood smears.

  5. Improving phosphate removal of sand infiltration system using alkaline fly ash.

    PubMed

    Cheung, K C; Venkitachalam, T H

    2000-07-01

    Septic tank effluent is customarily disposed of by soil infiltration. Coarse, sandy soil such as those found in Perth, Western Australia, exhibit low attenuation capabilities for phosphate (PO4(3-)) during effluent infiltration. Amendment of such soil with different amounts of alkaline precipitator and lagoon fly ashes was investigated as a means of reducing phosphorus (P) leakage to ground water. Alkaline precipitator fly ash possessed the highest P sorption capacity in terms of its Langmuir and Freundlich isotherm parameters during initial batch tests. The test materials were repeatedly contacted with fresh PO4(3-) solutions over 90 contacting cycles to gain a better indication of long-term P sorption capability. Again, precipitator fly ash exhibited higher P sorption capacity than lagoon fly ash and Spearwood sand. Column studies assessed the influence of various application rates of alkaline precipitator and lagoon fly ashes on the P removal of septic tank effluent. Septic tank effluent was applied at the rate of 4 cm/day to the column for 12 weeks. Concentrations of P were monitored in the column effluent. All the fly ash columns were more efficient in reducing P migration compared to the sand column. Increased levels of fly ash in the soil columns resulted in increased P attenuation. Lagoon fly ash was inferior to precipitator fly ash for P removal; high application rates of fly ash caused clogging of the infiltration bed apparently due to their lower permeability. It is reasoned that 5-15% precipitator fly ash, and less than 30% lagoon fly ash could be added to coarse sands to produce an infiltration bed, which would result in a better quality effluent than can be obtained with untreated sand alone.

  6. Sensitive terbium probes for luminescent determination of both alkaline phosphatase and codeine phosphate.

    PubMed

    Duerkop, Axel; Aleksandrova, Darya; Scripinets, Yuliya; Yegorova, Alla; Vityukova, Ekateryna

    2008-01-01

    New terbium complexes of 2-oxo-4-hydroxy-quinoline-3-carboxylic acid derivatives are reported for determination of alkaline phosphatase (aPase) and codeine phosphate (CP). Complexation with terbium occurs by way of the oxygen atoms of the 4-hydroxyl group and the carbonyl group of the 3-carboxy function, respectively. The complexes are highly luminescent and do not require luminescence enhancers. The excitation maxima of the terbium chelates are at 320 nm. The assay is based on the quenching of the hypersensitive 545-nm luminescence of the terbium complexes with the ligands 4-hydroxy-1-methyl-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid (5-ethyl-[1,3,4]thiadiazol-2-yl)-amide (L(1)) and 1-ethyl-4-hydroxy-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid (4-trifluoromethyl-phenyl)-amide (L(2)) by phosphate ions. The assays require equimolar (1.0 microM) concentrations of Tb(3+) and of the ligand at pH 8.0 (Tris-HCl buffer). The luminescence quenching is proportional to the concentration of aPase and CP within the range of 0.1-70 mU mL(-1) and 0.3-20 microg mL(-1), respectively. The detection limits were 0.05 mU mL(-1)=40.0 pmol L(-1) of aPase and 0.120 microg mL(-1) of CP.

  7. Zinc(II) oxide stability in alkaline sodium phosphate solutions at elevated temperatures

    SciTech Connect

    Ziemniak, S.E.; Opalka, E.P.

    1993-04-01

    Zinc oxide (ZnO) is shown to transform into either of two phosphate-containing compounds in relatively dilute alkaline sodium phosphate solutions at elevated temperatures via ZnO(s) + Na{sup +} + H{sub 2}PO{sub 4}{sup {minus}} {r_reversible} NaZnPO{sub 4}(s) + H{sub 2}O or 2 ZnO(s) + H{sub 3}PO{sub 4}(aq) {r_reversible} Zn{sub 2}(OH)PO{sub 4}(s) + H{sub 2}O. X-ray diffraction analyses indicate that NaZnPO{sub 4} possesses an orthorhombic unit cell having lattice parameters a = 8.710 {+-} 0.013, b = 15.175 {+-} 0.010, and c = 8.027 {+-} 0.004 {angstrom}. The thermodynamic equilibria for these reactions were defined in the system ZnO-Na{sub 2}O-P{sub 2}O{sub 5}-H{sub 2}O for Na/P molar ratios between 2.1 and 3. Based on observed reaction threshold values for sodium phosphate concentration and temperature, the standard entropy (S{degrees}) and free energy of formation ({Delta}G{sub f}{degrees}) for NaZnPO{sub 4} were calculated to be 169.0 J/mol-K and {minus}1510.6 kJ/mol, respectively; similar values for Zn{sub 2}(OH)PO{sub 4} (tarbuttite) were 235.9 J/mol-K and {minus}1604.6 kJ/mol. Additions of sodium sulfite and sulfate did not alter the above reactions.

  8. Alkaline aluminum phosphate glasses for thermal ion-exchanged optical waveguide

    NASA Astrophysics Data System (ADS)

    Wang, Fei; Chen, Baojie; Pun, Edwin Yue Bun; Lin, Hai

    2015-04-01

    Alkaline aluminum phosphate glasses (NMAP) with excellent chemical durability for thermal ion-exchanged optical waveguide have been designed and investigated. The transition temperature Tg (470 °C) is higher than the ion-exchange temperature (390 °C), which is favorable to sustain the stability of the glass structure for planar waveguide fabrication. The effective diffusion coefficient De of K+-Na+ ion exchange in NMAP glasses is 0.110 μm2/min, indicating that ion exchange can be achieved efficiently in the optical glasses. Single-mode channel waveguide has been fabricated on Er3+/Yb3+ doped NMAP glass substrate by standard micro-fabrication and K+-Na+ ion exchange. The mode field diameter is 9.6 μm in the horizontal direction and 6.0 μm in the vertical direction, respectively, indicating an excellent overlap with a standard single-mode fiber. Judd-Ofelt intensity parameter Ω2 is 5.47 × 10-20 cm2, implying a strong asymmetrical and covalent environment around Er3+ in the optical glasses. The full width at half maximum and maximum stimulated emission cross section of the 4I13/2 → 4I15/2 are 30 nm and 6.80 × 10-21 cm2, respectively, demonstrating that the phosphate glasses are potential glass candidates in developing compact optoelectronic devices. Pr3+, Tm3+ and Ho3+ doped NMAP glasses are promising candidates to fabricate waveguide amplifiers and lasers operating at special telecommunication windows.

  9. Alkaline phosphatase encapsulated in gellan-chitosan hybrid capsules.

    PubMed

    Fujii, Toshihiro; Ogiwara, Daisuke; Ohkawa, Kousaku; Yamamoto, Hiroyuki

    2005-05-23

    Alkaline phosphatase (ALP) was encapsulated in gellan-chitosan polyion complex (PIC) capsules using a convenient procedure. The recovery of ALP was about 50% when the capsules were prepared by dropping a solution of ALP and gellan mixture (ALP/gellan) into a chitosan solution. When p-nitrophenyl phosphate (p-NPP) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP) were incubated with ALP/gellan-chitosan capsules as substrates for ALP, the transparent colorless capsules changed to yellow and blue, respectively. The encapsulation of ALP into the PIC capsules was also confirmed by SDS-PAGE and immunoblot analyses. The ALP and polypeptides of more than 30 kDa remained without release even after incubation at 4 degrees C for 14 d. The biochemical properties of the encapsulated ALP activity were similar to those of the intact enzyme. When the solution containing p-NPP was loaded on a column packed with ALP/gellan-chitosan capsules at 27 degrees C, approximately 75% of p-NPP was hydrolyzed by passing through the column. No significant leakage of ALP was observed during the procedure, indicating that the capsules were resistant to pressure in the chromatographic operation. Furthermore, 70% of the hydrolytic activity of the packed capsules remained after storage at 4 degrees C for one month. These results suggest that the polyion complex capsules could be useful materials for protein fixation without chemical modification. [Diagram: see text] Encapsulation of ALP into PIC capsules and the morphological changes seen in the absence of the ALP substrate and in the presence of p-NPP and BICP.

  10. Rat serum alkaline phosphatase electrophoretic fractions: variations with feeding, starvation and cellulose fibre ingestion.

    PubMed

    Martins, M J; Dias, P O; Hipólito-Reis, C

    1998-12-01

    The effect of feeding, starvation and fibre ingestion on alkaline phosphatase (ALP) activity (E.C. 3.1.3.1) was studied in Wistar rat serum. Using identical assay conditions for total ALP activity determination and for electrophoretic ALP isoenzymes/fractions activity calculation, alpha- and beta-naphthyl phosphates and p-nitrophenyl phosphate were used as substrates and 2-amino-2-methyl-1-propanol/HCI was used as buffer, respectively. Total activity with beta-naphthyl phosphate was significantly higher than with alpha-naphthyl phosphate and p-nitrophenyl phosphate; with alpha-naphthyl phosphate it was significantly higher than with p-nitrophenyl phosphate. With all substrates, fed animals had significantly higher total activity than starving ones. Electrophoresis allowed the separation of two fractions. The second fraction activity was significantly higher in the fed group than in the starving ones, irrespective of the substrate used. Starving animals with fibre showed higher values of this fraction than starving animals without fibre, the difference reaching statistical significance with alpha-naphthyl phosphate. The first fraction predominated in both starved groups and the second in the fed group. The second fraction was identified as intestinal ALP. We conclude that the mechanical stimulation of the digestive tract appears to influence the passage of intestinal ALP to serum. The experimental conditions used enable quantification of electrophoretic fractions based on total activity. Activity depends on the substrate used.

  11. Rapid hydrolysis of model phosphate diesters by alkaline-earth cations in aqueous DMSO: speciation and kinetics.

    PubMed

    Taran, Olga; Medrano, Felipe; Yatsimirsky, Anatoly K

    2008-12-14

    Kinetics of the cleavage of two phosphate diesters, bis(4-nitrophenyl) phosphate and 2-hydroxypropyl 4-nitrophenyl phosphate and a triester, 4-nitrophenyl diphenyl phosphate, in the presence of Mg(II), Ca(II) and Sr(II) were studied in 90% vol. DMSO at 37 degrees C. The alkaline hydrolysis of the triester was inhibited by all cations, but with both phosphodiesters strong catalytic effects were observed. Potentiometric titrations of metal perchlorates by Bu4N(OH) revealed formation of M2(OH)3+, M(OH)+, M(OH)2 and M2(OH)5- species. Rate constants for phosphodiester cleavage by individual species were obtained from analysis of rate-concentration profiles. Observed first-order rate constants in the presence of 1-2 mM Mg(II) or Ca(II) in neutral and weakly basic solutions were 10(8)-10(11) times higher than those for background hydrolysis at the same pH while in water additions of up to 50 mM metal produced <100-fold accelerations. Possible structures of DMSO solvated catalyst-substrate complexes were modeled by DFT calculations with Mg(II). The increased catalytic activity in 90% DMSO is attributed to stronger association of hydroxide ions and anionic phosphodiesters with metal ions and to preferable solvation of cations by DMSO, which creates favorable for reaction anhydrous microenvironment in the coordination sphere of the catalyst.

  12. Lily pollen alkaline phytase is a histidine phosphatase similar to mammalian multiple inositol polyphosphate phosphatase (MINPP).

    PubMed

    Mehta, Bakul Dhagat; Jog, Sonali P; Johnson, Steven C; Murthy, Pushpalatha P N

    2006-09-01

    Phytic acid is the most abundant inositol phosphate in cells; it constitutes 1-5% of the dry weight of cereal grains and legumes. Phytases are the primary enzymes responsible for the hydrolysis of phytic acid and thus play important roles in inositol phosphate metabolism. A novel alkaline phytase in lily pollen (LlALP) was recently purified in our laboratory. In this paper, we describe the cloning and characterization of LlALP cDNA from lily pollen. Two isoforms of alkaline phytase cDNAs, LlAlp1 and LlAlp2, which are 1467 and 1533 bp long and encode proteins of 487 and 511 amino acids, respectively, were identified. The deduced amino acid sequences contains the signature heptapeptide of histidine phosphatases, -RHGXRXP-, but shares < 25% identity to fungal histidine acid phytases. Phylogenetic analysis reveals that LlALP is most closely related to multiple inositol polyphosphate phosphatase (MINPP) from humans (25%) and rats (23%). mRNA corresponding to LlAlp1 and LlAlp2 were expressed in leaves, stem, petals and pollen grains. The expression profiles of LlAlp isoforms in anthers indicated that mRNA corresponding to both isoforms were present at all stages of flower development. The expression of LlAlp2 cDNA in Escherichia coli revealed the accumulation of the active enzyme in inclusion bodies and confirmed that the cDNA encodes an alkaline phytase. In summary, plant alkaline phytase is a member of the histidine phosphatase family that includes MINPP and exhibits properties distinct from bacterial and fungal phytases.

  13. 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

  14. Inhibition of renal alkaline phosphatase by cimetidine.

    PubMed

    Minai-Tehrani, Dariush; Khodai, Somayeh; Aminnaseri, Somayeh; Minoui, Saeed; Sobhani-Damavadifar, Zahra; Alavi, Sana; Osmani, Raheleh; Ahmadi, Shiva

    2011-08-01

    Alkaline phosphatase (ALP) belongs to hydrolase group of enzymes. It is responsible for removing phosphate groups from many types of molecules, including nucleotides and proteins. Cimetidine (trade name Tagamet) is an antagonist of histamine H2-receptor that inhibits the production of gastric acid. Cimetidine is used for the treatment of gastrointestinal diseases. In this study the inhibitory effect of cimetidine on mouse renal ALP activity was investigated. Our results showed that cimetidine can inhibit ALP by uncompetitive inhibition. In the absence of inhibitor the V(max) and K(m) of the enzyme were found to be 13.7 mmol/mg prot.min and 0.25 mM, respectively. Both the Vmax and Km of the enzyme decreased with increasing cimetidine concentrations (0- 1.2 mM). The Ki and IC(50) of cimetidine were determined to be about 0.5 mM and 0.52 mM, respectively.

  15. 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)PO2 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.

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

    SciTech Connect

    Aisha, M.D.; Nor-Ashikin, M.N.K.; Sharaniza, A.B.R.; Nawawi, H.; Froemming, G.R.A.

    2015-09-10

    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 250 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.

  17. Effectiveness of recovered magnesium phosphates as fertilizers in neutral and slightly alkaline soils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Magnesium phosphates such as struvite (MgNH4PO4x6H2O) can be recovered from municipal, industrial and agricultural wastewaters. However, minimal research has been conducted on the beneficial reuse of these recovered products; conducted research has focused on low pH soils. This study determined wh...

  18. 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.

  19. 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.

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

    PubMed Central

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

    2017-01-01

    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. PMID:28181569

  1. Identification of alkaline phosphatase genes for utilizing a flame retardant, tris(2-chloroethyl) phosphate, in Sphingobium sp. strain TCM1.

    PubMed

    Takahashi, Shouji; Katanuma, Hiroshi; Abe, Katsumasa; Kera, Yoshio

    2017-03-01

    Tris(2-chloroethyl) phosphate (TCEP) is a haloalkyl phosphate flame retardant and plasticizer that has been recognized as a global environmental contaminant. Sphingobium sp. strain TCM1 can utilize TCEP as a phosphorus source. To identify the phosphomonoesterase involved in TCEP utilization, we identified four putative alkaline phosphatase (APase) genes, named SbphoA, SbphoD1, SbphoD2, and SbphoX-II, in the genome sequence. Following expression of these genes in Escherichia coli, APase activity was confirmed for the SbphoA and SbphoX-II gene products but was not clearly observed for the SbphoD1 and SbphoD2 gene products, owing to their accumulation in inclusion bodies. The single deletion of either SbphoA or SbphoX-II retarded the growth and reduced the APase activity of strain TCM1 cells on medium containing TCEP as the sole phosphorus source; these changes were more marked in cells with the SbphoX-II gene deletion. In contrast, the deletion of either SbphoD1 or SbphoD2 had no effect on cell growth or APase activity. The double deletion of SbphoA and SbphoX-II resulted in the complete loss of cell growth on TCEP. These results show that SbPhoA and SbPhoX-II are involved in the utilization of TCEP as a phosphorus source and that SbPhoX-II is the major phosphomonoesterase involved in TCEP utilization.

  2. Ratiometric fluorescent probe for alkaline phosphatase based on betaine-modified polyethylenimine via excimer/monomer conversion.

    PubMed

    Zheng, Fangyuan; Guo, Sihua; Zeng, Fang; Li, Jun; Wu, Shuizhu

    2014-10-07

    Alkaline phosphatase (ALP) is an important diagnostic indicator for a number of human diseases since abnormal level of ALP is closely related to a variety of pathological processes; hence, the development of convenient and reliable assay methods for monitoring ALP is of great significance for medical sciences as well as biological diagnostics. Herein, we report the first ratiometric fluorescent sensing system for ALP. This sensing system consists of two components: the betaine-modified and positively charged polyethylenimine (PEI) and the negatively charged pyrene derivative containing one ALP-responsive phosphate group (Py-P, an aliphatic phosphate ester). In the absence of ALP, the two-component sensing system shows the excimer's emission of Py-P, since Py-P molecules complex with the positively charged polyelectrolyte via electrostatic interactions, leading to the formation of pyrene excimers. While in the presence of ALP, the phosphate moieties are cleaved from Py-P molecules due to the enzymatic reaction, thereby destroying the electrostatic interactions; as a result, the system displays the monomer emission of Py-P. This assay system is operable in aqueous media with a very low detection limit of 0.1 U/mL. The system is capable of detecting ALP in such biological fluid as serum, and this strategy may provide a new and effective approach for designing ratiometric sensing systems for detecting other biomolecules.

  3. 4-Nitrophenol in 4-nitrophenyl phosphate, a substrate for alkaline phosphatase, as measured by paired-ion high-performance liquid chromatography.

    PubMed

    Culbreth, P H; Duncan, I W; Burtis, C A

    1977-12-01

    We used paired-ion high-performance liquid chromatography to determine the 4-nitrophenol content of 4-nitrophenyl phosphate, a substrate for alkaline phosphatase analysis. This was done on a reversed-phase column with a mobile phase of methanol/water, 45/55 by vol, containing 3 ml of tetrabutylammonium phosphate reagent per 200 ml of solvent. At a flow rate of 1 ml/min, 4-nitrophenol was eluted at 9 min and monitored at 404 nm; 4-nitrophenyl phosphate was eluted at 5 min and could be monitored at 311 nm. Samples of 4-nitrophenyl phosphate obtained from several sources contained 0.3 to 7.8 mole of 4-nitrophenol per mole of 4-nitrophenyl phosphate.

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

    PubMed

    Aisha, M D; Nor-Ashikin, M N K; Sharaniza, A B R; Nawawi, H; Froemming, G R A

    2015-09-10

    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 250 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.

  5. Acid and alkaline phosphatases of Capnocytophaga species. II. Isolation, purification, and characterization of the enzymes from Capnocytophaga ochracea.

    PubMed

    Poirier, T P; Holt, S C

    1983-10-01

    Capnocytophaga ochracea acid (AcP; EC 3.1.3.2) and alkaline (AlP; EC 3.1.3.1) phosphatase was isolated by Ribi cell disruption and purified by sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE.) Both phosphatases eluted from Sephadex G-150 consistent with molecular weights (migration) of 140 000 and 110 000. SDS-PAGE demonstrated a 72 000 and 55 000 subunit molecular migration for AcP and AlP, respectively. The kinetics of activity of purified AcP and AlP on p-nitrophenol phosphate and phosphoseryl residues of the phosphoproteins are presented.

  6. Glucosamine: fructose-6-phosphate amidotransferase in the white shrimp Litopenaeus vannamei: characterization and regulation under alkaline and cadmium stress.

    PubMed

    Liu, Y; Cai, D X; Wang, L; Li, J Z; Wang, W N

    2015-10-01

    Heavy metal residues and chemical contaminators considered as relevant sources of aquatic environmental pollutants have a generally immunosuppressive effect on aquatic organisms, depressing metabolic activities and immune response. Glutamine: fructose-6-phosphate aminotransferase (GFAT, EC2.6.1.16) is the first, and rate-limiting, enzyme in the hexosamine biosynthetic pathway, and is involved in the regulation of chitin biosynthesis and glycosylation of proteins. We have isolated and characterized GFAT from the white shrimp Litopenaeus vannamei. Amino acid sequence similarity of the Lv-GFAT (L.vannamei-GFAT) was highest to GFATs isolated from insects and mammals (83 % similarity to that of Haemaphysalis longicornis). The open-reading frame of the Lv-GFAT codes for a protein of 41.6 kDa with a calculated isoelectric point of 5.03. RT-PCR assays showed that endogenous Lv-GFAT mRNA is most strongly expressed in the intestine. Further analysis of Lv-GFAT gene expression in hepatopancreas by quantitative real-time PCR demonstrated that Lv-GFAT transcript levels increased when the shrimp were exposed to alkaline pH (9.3) and cadmium stress, but the time when its mRNA expression level peaked differed under these stresses. We also first expressed the recombinant protein of GFAT from shrimps in Escherichia coli. Western blot analyses confirmed that the Lv-GFAT protein was strongly expressed in the hepatopancreas after exposure to the LC-Cd stress. These results suggest that Lv-GFAT expression is stimulated by alkaline pH and cadmium stress and that it may play important roles in resistance of shrimp to environmental stresses.

  7. Heterologous expression and functional characterization of a plant alkaline phytase in Pichia pastoris.

    PubMed

    Johnson, Steven C; Yang, Mimi; Murthy, Pushpalatha P N

    2010-12-01

    Phytases catalyze the sequential hydrolysis of phytic acid (myo-insositol hexakisphosphate), the most abundant inositol phosphate in cells. Phytic acid constitutes 3-5% of the dry weight of cereal grains and legumes such as corn and soybean. The high concentration of phytates in animal feed and the inability of non-ruminant animals such as swine and poultry to digest phytates leads to phosphate contamination of soil and water bodies. The supplementation of animal feed with phytases results in increased bioavailability to animals and decreased environmental contamination. Therefore, phytases are of great commercial importance. Phytases with a range of properties are needed to address the specific digestive needs of different animals. Alkaline phytase (LlALP1 and LlALP2) which possess unique catalytic properties that have the potential to be useful as feed and food supplement has been identified in lily pollen. Substantial quantities of alkaline phytase are needed for animal feed studies. In this paper, we report the heterologous expression of LlALP2 from lily pollen in Pichia pastoris. The expression of recombinant LlALP2 (rLlALP2) was optimized by varying the cDNA coding for LlALP2, host strain and growth conditions. The catalytic properties of recombinant LlALP2 were investigated extensively (substrate specificity, pH- and temperature dependence, and the effect of Ca(2+), EDTA and inhibitors) and found to be very similar to that of the native LlALP2 indicating that rLlALP2 from P. pastoris can serve as a potential source for structural and animal feed studies.

  8. 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.

  9. 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

  10. Kinetics of the alkaline phosphatase catalyzed hydrolysis of disodium p-nitrophenyl phosphate in frozen model systems.

    PubMed

    Terefe, Netsanet Shiferaw; Mokwena, Kereilemang Khanah; Loey, Ann Van; Hendrickx, Marc E

    2002-01-01

    The alkaline phosphatase catalyzed hydrolysis of disodium-p-nitrophenyl phosphate was studied in four model systems comprising sucrose, maltodextrin, carboxymethylcellulose (CMC), and CMC-lactose in a temperature range of -28 to 20 degrees C. In the maltodextrin and CMC-lactose model systems, the reaction rate decreased to a very low value as the glass transition temperature was approached. In the CMC and CMC-lactose systems with low initial solute concentration, as a consequence of freeze-concentration, a rate maximum around the initial freezing temperature was observed. The Arrhenius equation described the temperature dependence of the reaction rate both in the liquid and the glassy states in all systems studied, while a slightly curved Arrhenius plot was observed in the "rubbery" state of the CMC and CMC-lactose systems. The WLF equation with system-dependent coefficients described the kinetics in the rubbery state of all the model systems except sucrose, excluding the short temperature range where reaction rate enhancement with decreasing temperature was observed.

  11. 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

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

    PubMed

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

    2016-01-01

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

  13. Characterization of the Proteomic Profiles of the Brown Tide Alga Aureoumbra lagunensis under Phosphate- and Nitrogen-Limiting Conditions and of Its Phosphate Limitation-Specific Protein with Alkaline Phosphatase Activity

    PubMed Central

    Sun, Ming-Ming; Sun, Jin; Qiu, Jian-Wen; Jing, Hongmei

    2012-01-01

    The persistent bloom of the brown tide alga Aureoumbra lagunensis has been reported in coastal embayments along southern Texas, but the molecular mechanisms that sustain such algal bloom are unknown. We compared the proteome and physiological parameters of A. lagunensis grown in phosphate (P)-depleted, P- and nitrogen (N)-depleted, and nutrient-replete cultures. For the proteomic analysis, samples from three conditions were subjected to two-dimensional electrophoresis and tandem mass spectrometry analysis. Because of the paucity of genomic resources in this species, a de novo cross-species protein search was used to identify the differentially expressed proteins, which revealed their involvement in several key biological processes, such as chlorophyll synthesis, antioxidative protection, and protein degradation, suggesting that A. lagunensis may adopt intracellular nutrient compensation, extracellular organic nutrient regeneration, and damage protection to thrive in P-depleted environments. A highly abundant P limitation-specific protein, tentatively identified as a putative alkaline phosphatase, was further characterized by enzyme activity assay on nondenaturing gel and confocal microscopy, which confirmed that this protein has alkaline phosphatase activity, is a cytoplasmic protein, and is closely associated with the cell membrane. The abundance, location, and functional expression of this alkaline phosphatase all indicate the importance of organic P utilization for A. lagunensis under P limitation and the possible role of this alkaline phosphatase in regenerating phosphate from extra- or intracellular organic phosphorus. PMID:22247172

  14. 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.

  15. 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.

  16. 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

    2017-03-08

    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.

  17. 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.

  18. Alkaline phosphatase based amperometric biosensor immobilized by cysteamine-glutaraldehyde modified self-assembled monolayer.

    PubMed

    Yorganci, Emine; Akyilmaz, Erol

    2011-10-01

    Alkaline phosphatase (ALP) was immobilized with cross-linking agents glutaraldehyde and cysteamine by forming a self-assembled monolayer on a screen printed gold electrode. ALP converts p-nitrophenyl phosphate to p-nitrophenol and phosphate. p-Nitrophenol loses H(+) ion and turns into the negatively charged compound p-nitrophenolate at medium pH. As a result, the unstable product formed is measured chronoamperometrically at an application potential of + 0.95 V. The biosensor response depends linearly on p-nitrophenyl phosphate concentration between 0.05 - 0.6 mM with a response time of 40 seconds. Detection limit of the biosensor is 0.033 mM.

  19. Ingestion of potato starch containing esterified phosphorus increases alkaline phosphatase activity in the small intestine in rats.

    PubMed

    Mineo, Hitoshi; Morikawa, Nao; Ohmi, Sayako; Ishida, Kyo; Machida, Ayaka; Kanazawa, Takumi; Chiji, Hideyuki; Fukushima, Michihiro; Noda, Takahiro

    2010-05-01

    Alkaline phosphatase (ALP) hydrolyzes a variety of monophosphate esters and plays an important role in phosphorus (P) metabolism. Several nutrients in food have been reported to affect intestinal ALP activity in animal models. Previous reports indicated that high levels of P or phosphate in diets decreased intestinal ALP activity in rats. Because potato starch contains considerable amounts of esterified P, unlike other starch-derived plants, we hypothesized that the feeding of potato starch would decrease ALP activity in the intestinal tract. Male Sprague-Dawley rats (7 weeks old) were fed 3 different types of diet containing 60% corn starch or 1 of 2 types of potato starch with different esterified P content for 1 or 5 weeks. Body weight and food intake of each rat were measured every day throughout the experimental periods. At the end of the feeding periods, the small intestine was removed to determine ALP activity in the mucosal tissues. Significant differences were observed in ALP activity in the small intestine between the 2 feeding periods, among the 4 segments of the small intestine, and among the 3 diet groups. Significant positive linear correlations between the amount of P derived from the starch and mucosal ALP activity were obtained in the jejunum and jejunoileum in rats after feeding for 5 weeks. We concluded, contrary to our hypotheses, that the ingestion of potato starch adaptively increases ALP activity in the upper part of the small intestine of growing rats in an esterified P content-dependent manner.

  20. 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

  1. 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.

  2. Kinetics of calcium phosphate nucleation and growth on calcite: implications for predicting the fate of dissolved phosphate species in alkaline soils.

    PubMed

    Wang, Lijun; Ruiz-Agudo, Encarnación; Putnis, Christine V; Menneken, Martina; Putnis, Andrew

    2012-01-17

    Unraveling the kinetics of calcium orthophosphate (Ca-P) precipitation and dissolution is important for our understanding of the transformation and mobility of dissolved phosphate species in soils. Here we use an in situ atomic force microscopy (AFM) coupled with a fluid reaction cell to study the interaction of phosphate-bearing solutions with calcite surfaces. We observe that the mineral surface-induced formation of Ca-P phases is initiated with the aggregation of clusters leading to the nucleation and subsequent growth of Ca-P phases on calcite, at various pH values and ionic strengths relevant to soil solution conditions. A significant decrease in the dissolved phosphate concentration occurs due to the promoted nucleation of Ca-P phases on calcite surfaces at elevated phosphate concentrations and more significantly at high salt concentrations. Also, kinetic data analyses show that low concentrations of citrate caused an increase in the nucleation rate of Ca-P phases. However, at higher concentrations of citrate, nucleation acceleration was reversed with much longer induction times to form Ca-P nuclei. These results demonstrate that the nucleation-modifying properties of small organic molecules may be scaled up to analyze Ca-P dissolution-precipitation processes that are mediated by a more complex soil environment. This in situ observation, albeit preliminary, may contribute to an improved understanding of the fate of dissolved phosphate species in diverse soil systems.

  3. 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 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.

  4. 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.

  5. Osteoregenerative capacities of dicalcium phosphate-rich calcium phosphate bone cement.

    PubMed

    Ko, Chia-Ling; Chen, Jian-Chih; Tien, Yin-Chun; Hung, Chun-Cheng; Wang, Jen-Chyan; Chen, Wen-Cheng

    2015-01-01

    Calcium phosphate cement (CPC) is a widely used bone substitute. However, CPC application is limited by poor bioresorption, which is attributed to apatite, the stable product. This study aims to systematically survey the biological performance of dicalcium phosphate (DCP)-rich CPC. DCP-rich CPC exhibited a twofold, surface-modified DCP anhydrous (DCPA)-to-tetracalcium phosphate (TTCP) molar ratio, whereas conventional CPC (c-CPC) showed a onefold, surface unmodified DCPA-to-TTCP molar ratio. Cell adhesion, morphology, viability, and alkaline phosphatase (ALP) activity in the two CPCs were examined with bone cell progenitor D1 cultured in vitro. Microcomputed tomography and histological observation were conducted after CPC implantation in vivo to analyze the residual implant ratio and new bone formation rate. D1 cells cultured on DCP-rich CPC surfaces exhibited higher cell viability, ALP activity, and ALP quantity than c-CPC. Histological evaluation indicated that DCP-rich CPC showed lesser residual implant and higher new bone formation rate than c-CPC. Therefore, DCP-rich CPC can improve bioresorption. The newly developed DCP-rich CPC exhibited potential therapeutic applications for bone reconstruction.

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

    NASA Astrophysics Data System (ADS)

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

    2016-01-01

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

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

    PubMed

    Douglas, Timothy E L; Skwarczynska, Agata; Modrzejewska, Zofia; Balcaen, Lieve; Schaubroeck, David; Lycke, Sylvia; Vanhaecke, Frank; Vandenabeele, Peter; Dubruel, Peter; Jansen, John A; Leeuwenburgh, Sander C G

    2013-05-01

    Thermosensitive chitosan hydrogels containing sodium beta-glycerophosphate (β-GP), whose gelation is induced by increasing temperature to body temperature, were functionalized by incorporation of alkaline phosphatase (ALP), an enzyme involved in mineralization of bone. ALP incorporation led to acceleration of gelation upon increase of temperature for four different chitosan preparations of differing molecular weight, as demonstrated by rheometric time sweeps at 37 °C. Hydrogels containing ALP were subsequently incubated in calcium glycerophosphate (Ca-GP) solution to induce their mineralization with calcium phosphate (CaP) in order to improve their suitability as materials for bone replacement. Incorporated ALP retained its bioactivity and induced formation of CaP mineral, as confirmed by SEM, FTIR, Raman spectroscopy, XRD, ICP-OES, and increases in dry mass percentage, which rose with increasing ALP concentration and incubation time in Ca-GP solution. The results demonstrate that ALP accelerates formation of thermosensitive chitosan/β-GP hydrogels and induces their mineralization with CaP, which paves the way for applications as injectable bone replacement materials.

  8. The effects of retinoic acid on alkaline phosphatase activity and tissue-non-specific alkaline phosphatase gene expression in human periodontal ligament cells and gingival fibroblasts.

    PubMed

    San Miguel, S M; Goseki-Sone, M; Sugiyama, E; Watanabe, H; Yanagishita, M; Ishikawa, I

    1998-10-01

    Alkaline phosphatase (ALP) in human periodontal ligament (HPDL) cells is classified as a tissue-non-specific alkaline phosphatase (TNSALP) by its enzymatic and immunological properties. Since retinoic acid (RA) has been shown as a potent inducer of TNSALP expression in various osteoblastic and fibroblastic cells, we investigated the effects of RA on the level of ALP activity and expression of TNSALP mRNAs in HPDL cells. Cultured cells were treated with desired RA concentrations (0, 10(-7), 10(-6), 10(-5) M) in medium containing 1% bovine serum albumin without serum. ALP activity was determined by the rate of hydrolysis of p-nitrophenyl phosphate and was also assayed in the presence of specific inhibitors. In order to identify the TNSALP mRNA type expressed by HPDL, a set of oligonucleotide primers corresponding to 2 types of human TNSALP mRNA (i.e. bone-type and liver-type) were designed, and mRNA isolated from HPDL was amplified by means of reverse transcription-polymerase chain reaction (RT-PCR). After treatment with RA (10(-6) M) for 4 d, there was a significant increase in the ALP activity of HPDL cells. The use of inhibitors and thermal inactivation experiments showed that the increased ALP activity had properties of the TNSALP type. RT-PCR analysis revealed that bone-type mRNA was highly stimulated in HPDL cells by RA treatment, but the expression of liver-type mRNA was not detected. These results indicated that the upregulation of ALP activity in HPDL cells by RA was due to the increased transcription of bone-type mRNA of the TNSALP gene.

  9. 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.

  10. Structure of iron phosphate glasses modified by alkali and alkaline earth additions: neutron and x-ray diffraction studies.

    PubMed

    Bingham, P A; Barney, E R

    2012-05-02

    The structure of iron phosphate glasses modified by additions of K(2)O and BaO, with nominal molar compositions [(1 - x)(0.6P(2)O(5)-0.4Fe(2)O(3))]xMe(y)O, where x = 0-0.4 in increments of 0.1; Me=K or Ba; and y = 1 or 2, has been investigated using neutron diffraction and x-ray diffraction techniques. Fitted coordination numbers for P-O and Fe-O showed a notable change in the P-O(NBO) and P-O(BO) contributions at greater than 20 mol% modifier addition, with barium producing a markedly larger increase in P-O(NBO) contribution than potassium. Fitting of T(N)(r) and T(X)(r) provided average n(BaO) = 9 and n(KO) = 6. Iron occurs in a range of coordination sites in these glasses: ([6])Fe(2+), ([4])Fe(3+), ([5])Fe(3+) and ([6])Fe(3+). The partitioning between these sites gives average n(FeO) = 5.2-5.8, with barium-doped glasses exhibiting higher average n(FeO) than potassium-doped glasses. The stronger depolymerizing effect of Ba(2+) than K(+) on the phosphate network, coupled with its greater ionic charge and higher Me-O, Fe-O and O···O coordination numbers, explain previously observed divergences in physical properties between the barium-doped and the potassium-doped glasses.

  11. [Changes of serum alkaline phosphatase and electrolytes during 21 d head down bed-rest].

    PubMed

    Yao, Yong-jie; Sun, Xi-qing; Wang, Zhong-bo; Zhao, Shuang-bao; Yang, Chang-bin; Wu, Xing-yu

    2002-06-01

    Objective. To investigate the effect of simulated weightlessness on serum alkaline phosphatase (ALP), calcium, magnesium, chlorine and phosphorus. Method. 6 healthy males, aged 24.8 +/- 6.1, were exposed to -6 degrees HDT bed rest for 21 d. Activity of serum alkaline phosphatase, serum contents of calcium (Ca), magnesium (Mg), chlorine (Cl) and phosphorus (P) ions were assayed before HDT (d-3), on the 3rd, 10th and 21st day during HDT and after HDT (d+2). Ca was measured by methyl thymol blue method, P was determined with ultraviolet spectrophotography, determination of Mg and Cl were made with enzyme method, ALP was examined with 4-nitrobenzene phosphate method. Result. Serum Ca2+ levels were significantly higher at d10, d2l and d+2 than the value of d-3 (P<0.01). P3+ levels declined significantly on d2l as compared with d-3 (P<0.01). During the HDT and after HDT, Mg2+ declined to a level below that before HDT (P<0.05 or P<0.01). Cl- were significantly higher at d2l and d+2 than the value of d-3 (P<0.01). ALP level was higher on d2l than on d-3 (P<0.01). Conclusion. 21 d HDT induced increase of Ca, Cl, ALP, and decline of Mg and P. The changes may reflect the imbalance of metabolism.

  12. Kinetics of the alkaline phosphatase catalyzed hydrolysis of disodium p-nitrophenyl phosphate: effects of carbohydrate additives, low temperature, and freezing.

    PubMed

    Terefe, Netsanet Shiferaw; Arimi, Joshua Mbaabu; Van Loey, Ann; Hendrickx, Marc

    2004-01-01

    The kinetics of the alkaline phosphatase catalyzed hydrolysis of disodium p-nitrphenyl phosphate was studied at 25 degrees C in the presence of the carbohydrates sucrose, fructose, lactose, maltodextrin (DE = 13-17), carboxymethylcellulose (CMC), and CMC-lactose (in 1:1 proportion) at different concentrations and in the presence of sucrose at two different concentrations in a temperature range between 25 and -10 degrees C in subcooled and frozen systems. The objective was to determine whether the reaction is diffusion-controlled, to gain an insight about the factors that determine the diffusion of the reaction species, to understand the mechanism through which the different carbohydrate additives affect the kinetics of the reaction, and to determine the effect of low temperature and freezing on the structural conformation of the enzyme. It was found that the alkaline phosphatase catalyzed hydrolysis of DNPP under the condition studied is at least partially diffusion-controlled. The results also indicate that the diffusion is not controlled by the macroviscosity of the reaction media. The concentration and type of the molecules that constitute the background matrix seem to be the main factors governing the reaction. The results indicate that the different carbohydrates affect the kinetics of the reaction through the excluded volume effect of molecular crowding and decreased substrate and product diffusion rate and not through nonspecific solute effects, which may cause protein denaturation and alteration in enzyme activity. Low temperature does not seem to affect the structural conformation of the enzyme in the temperature range studied, whereas freezing affected the catalytic properties of the enzyme perhaps through its effect on the structural conformation of the enzyme.

  13. 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.

  14. Inhibition of dsDNA-templated copper nanoparticles by pyrophosphate as a label-free fluorescent strategy for alkaline phosphatase assay.

    PubMed

    Zhang, Liangliang; Zhao, Jingjin; Duan, Min; Zhang, Hua; Jiang, Jianhui; Yu, Ruqin

    2013-04-16

    On the basis of the inhibition of double strand DNA (dsDNA)-templated fluorescent copper nanoparticles (CuNPs) by pyrophosphate (PPi), a novel label-free turn-on fluorescent strategy to detect alkaline phosphatase (ALP) under physiological conditions has been developed. This method relies on the strong interaction between PPi and Cu(2+), which would hamper the effective formation of fluorescent CuNPs, leading to low fluorescence intensity. The ALP-catalyzed PPi hydrolysis would disable the complexation between Cu(2+) and PPi, facilitating the formation of fluorescent CuNPs through the reduction by ascorbate in the presence of dsDNA templates. Thus, the fluorescence intensity was recovered, and the fluorescence enhancement was related to the concentration of ALP. This method is cost-effective and convenient without any labels or complicated operations. The present strategy exhibits a high sensitivity and the turn-on mode provides a high selectivity for the ALP assay. Additionally, the inhibition effect of phosphate on the ALP activity was also studied. The proposed method using a PPi substrate may hold a potential application in diagnosis of ALP-related diseases or evaluation of ALP functions in biological systems.

  15. ALPs at colliders

    NASA Astrophysics Data System (ADS)

    Mimasu, Ken; Sanz, Verónica

    2015-06-01

    New pseudo-scalars, often called axion-like particles (ALPs), abound in model-building and are often associated with the breaking of a new symmetry. Traditional searches and indirect bounds are limited to light axions, typically in or below the KeV range for ALPs coupled to photons. We present collider bounds on ALPs from mono-γ, tri-γ and mono-jet searches in a model independent fashion, as well as the prospects for the LHC and future machines. We find that they are complementary to existing searches, as they are sensitive to heavier ALPs and have the capability to cover an otherwise inaccessible region of parameter space. We also show that, assuming certain model dependent correlations between the ALP coupling to photons and gluons as well as considering the validity of the effective description of ALP interactions, mono-jet searches are in fact more suitable and effective in indirectly constraining ALP scenarios.

  16. Colocation and role of polyphosphates and alkaline phosphatase in apatite biomineralization of elasmobranch tesserae.

    PubMed

    Omelon, Sidney; Georgiou, John; Variola, Fabio; Dean, Mason N

    2014-09-01

    Elasmobranchs (e.g. sharks and rays), like all fishes, grow continuously throughout life. Unlike other vertebrates, their skeletons are primarily cartilaginous, comprising a hyaline cartilage-like core, stiffened by a thin outer array of mineralized, abutting and interconnected tiles called tesserae. Tesserae bear active mineralization fronts at all margins and the tesseral layer is thin enough to section without decalcifying, making this a tractable but largely unexamined system for investigating controlled apatite mineralization, while also offering a potential analog for endochondral ossification. The chemical mechanism for tesserae mineralization has not been described, but has been previously attributed to spherical precursors, and alkaline phosphatase (ALP) activity. Here, we use a variety of techniques to elucidate the involvement of phosphorus-containing precursors in the formation of tesserae at their mineralization fronts. Using Raman spectroscopy, fluorescence microscopy and histological methods, we demonstrate that ALP activity is located with inorganic phosphate polymers (polyP) at the tessera-uncalcified cartilage interface, suggesting a potential mechanism for regulated mineralization: inorganic phosphate (Pi) can be cleaved from polyP by ALP, thus making Pi locally available for apatite biomineralization. The application of exogenous ALP to tissue cross-sections resulted in the disappearance of polyP and the appearance of Pi in uncalcified cartilage adjacent to mineralization fronts. We propose that elasmobranch skeletal cells control apatite biomineralization by biochemically controlling polyP and ALP production, placement and activity. Previous identification of polyP and ALP shown previously in mammalian calcifying cartilage supports the hypothesis that this mechanism may be a general regulating feature in the mineralization of vertebrate skeletons.

  17. Phosphate taxis in Pseudomonas aeruginosa.

    PubMed

    Kato, J; Ito, A; Nikata, T; Ohtake, H

    1992-08-01

    Pseudomonas aeruginosa was shown to be attracted to phosphate. The chemotactic response was induced by phosphate starvation. The specificity of chemoreceptors for phosphate was high so that no other tested phosphorus compounds elicited a chemotactic response as strong as that elicited by phosphate. Competition experiments showed that the chemoreceptors for phosphate appeared to be different from those for the common amino acids. Mutants constitutive for alkaline phosphatase showed the chemotactic response to phosphate regardless of whether the cells were starved for phosphate.

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

    PubMed

    Ball, Vincent

    2014-09-01

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

  19. 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.

  20. Electrochemical sandwich-type biosensors for α-1 antitrypsin with carbon nanotubes and alkaline phosphatase labeled antibody-silver nanoparticles.

    PubMed

    Zhu, Gangbing; Lee, Hye Jin

    2017-03-15

    A novel sandwich-type biosensor was developed for the electrochemical detection of α-1 antitrypsin (AAT, a recognized biomarker for Alzheimer's disease). The biosensor was composed of 3, 4, 9, 10-perylene tetracarboxylic acid/carbon nanotubes (PTCA-CNTs) as a sensing platform and alkaline phosphatase-labeled AAT antibody functionalized silver nanoparticles (ALP-AAT Ab-Ag NPs) as a signal enhancer. CNTs offer high surface area and good electrical conductivity. Importantly, Ag NPs could increase the amount of ALP on the sensing surface and the ALP could dephosphorylate 4-amino phenyl phosphate (APP) enzymatically to produce electroactive species 4-aminophenol (AP). For detecting AAT based on the sandwich-type biosensor, the results show that the peak current value of AP using ALP-AAT Ab-Ag NPs as signal enhancer is much higher than that by using ALP-AAT Ab bioconjugate (without Ag NPs), the biosensor exhibited desirable performance for AAT determination with a wide linearity in the range from 0.05 to 20.0pM and a low detection limit of 0.01pM. Finally, the developed sensor was successfully applied to the analysis of AAT concentration in serum samples.

  1. Fabrications of zinc-releasing biocement combining zinc calcium phosphate to calcium phosphate cement.

    PubMed

    Horiuchi, Shinya; Hiasa, Masahiro; Yasue, Akihiro; Sekine, Kazumitsu; Hamada, Kenichi; Asaoka, Kenzo; Tanaka, Eiji

    2014-01-01

    Recently, zinc-releasing bioceramics have been the focus of much attention owing to their bone-forming ability. Thus, some types of zinc-containing calcium phosphate (e.g., zinc-doped tricalcium phosphate and zinc-substituted hydroxyapatite) are examined and their osteoblastic cell responses determined. In this investigation, we studied the effects of zinc calcium phosphate (ZCP) derived from zinc phosphate incorporated into calcium phosphate cement (CPC) in terms of its setting reaction and MC3T3-E1 osteoblast-like cell responses. Compositional analysis by powder X-ray diffraction analysis revealed that HAP crystals were precipitated in the CPC containing 10 or 30wt% ZCP after successfully hardening. However, the crystal growth observed by scanning electron microscopy was delayed in the presence of additional ZCP. These findings indicate that the additional zinc inhibits crystal growth and the conversion of CPC to the HAP crystals. The proliferation of the cells and alkaline phosphatase (ALP) activity were enhanced when 10wt% ZCP was added to CPC. Taken together, ZCP added CPC at an appropriate fraction has a potent promotional effect on bone substitute biomaterials.

  2. Conjugated polyelectrolyte-based real-time fluorescence assay for alkaline phosphatase with pyrophosphate as substrate.

    PubMed

    Liu, Yan; Schanze, Kirk S

    2008-11-15

    The fluorescence of the anionic, carboxylate-substituted poly(phenylene ethynylene) polymer PPECO2 is quenched very efficiently via the addition of 1 equiv of Cu(2+). Addition of pyrophosphate (PPi) into the weakly fluorescent solution of PPECO2 and Cu(2+) induces recovery of the polymer's fluorescence; the recovery occurs because PPi complexes with Cu(2+), effectively sequestering the ion so it cannot bind to the carboxylate groups of the polymer. A calibration curve was developed that relates the extent of fluorescence recovery to [PPi], making the PPECO2-Cu(2+) system a sensitive and selective turn-on sensor for PPi. Using the PPECO2-Cu(2+) system as the signal transducer, a real-time fluorescence turn-off assay for the enzyme alkaline phosphatase (ALP) using PPi as the substrate is developed. The assay operates with [PPi] in the micromolar range, and it offers a straightforward and rapid detection of ALP activity with the enzyme present in the nanomolar concentration range, operating either in an end point or real-time format. Kinetic and product inhibition parameters are derived by converting time-dependent fluorescence intensity into PPi (substrate) concentration, thus allowing calculation of the initial reaction rates (v(o)). Weak, nonspecific fluorescence responses are observed concomitant to addition of other proteins to the assay solution; however, the signal response to ALP is demonstrated to arise from the ALP catalyzed hydrolysis of PPi to phosphate (Pi).

  3. Light triggered detection of aminophenyl phosphate with a quantum dot based enzyme electrode

    PubMed Central

    2011-01-01

    An electrochemical sensor for p-aminophenyl phosphate (pAPP) is reported. It is based on the electrochemical conversion of 4-aminophenol (4AP) at a quantum dot (QD) modified electrode under illumination. Without illumination no electron transfer and thus no oxidation of 4AP can occur. pAPP as substrate is converted by the enzyme alkaline phosphatase (ALP) to generate 4AP as a product. The QDs are coupled via 1,4-benzenedithiol (BDT) linkage to the surface of a gold electrode and thus allow potential-controlled photocurrent generation. The photocurrent is modified by the enzyme reaction providing access to the substrate detection. In order to develop a photobioelectrochemical sensor the enzyme is immobilized on top of the photo-switchable layer of the QDs. Immobilization of ALP is required for the potential possibility of spatially resolved measurements. Geometries with immobilized ALP are compared versus having the ALP in solution. Data indicate that functional immobilization with layer-by-layer assembly is possible. Enzymatic activity of ALP and thus the photocurrent can be described by Michaelis- Menten kinetics. pAPP is detected as proof of principle investigation within the range of 25 μM - 1 mM. PMID:21982200

  4. Sensitive and selective colorimetric assay of alkaline phosphatase activity with Cu(II)-phenanthroline complex.

    PubMed

    Hu, Qiong; He, Minhui; Mei, Yaqi; Feng, Wenjie; Jing, Su; Kong, Jinming; Zhang, Xueji

    2017-01-15

    Alkaline phosphatase (ALP) plays a vital role in dephosphorylation- and phosphorylation-related cellular regulation and signaling processes. Accordingly, the development of efficient methods for ALP activity assay is of significant importance in clinical diagnosis. In this work, a simple and practical method is reported for the first time for the sensitive and selective colorimetric assay of ALP activity by exploiting a water-soluble Cu(II)-phenanthroline complex as the probe, on the basis of the distinctive metal-to-ligand charge-transfer (MLCT) properties. This method is simply built on a two-step chromogenic reaction: the enzymatic hydrolysis of the substrate ascorbic acid 2-phosphate to ascorbic acid (AA), followed by the reduction of the colorimetric probe Cu(BPDS)2(2-) (BPDS=bathophenanthroline disulfonate) by AA to its cuprous form. The latter process triggers a turn-on spectral absorption at 424nm and a striking color change of the solution from colorless to blackish-green. Needless of complicated protocols and instrumentation, this method allows a sensitive readout of ALP activity within a wide linear range of 0-200mUmL(-)(1), with a detection limit down to 1.25mUmL(-1). Results also reveal that it is highly selective and holds great potential in ALP inhibitor efficiency evaluation. In addition, quantitative analysis of ALP activity in spiked serum samples has been realized successfully in the linear range of 0-200mUmL(-1), with a detection limit of 1.75mUmL(-1). Advantages of simplicity, wide linear range, high sensitivity and selectivity, low cost, and little background interference render this method great potential in practical applications.

  5. Facile colorimetric assay of alkaline phosphatase activity using Fe(II)-phenanthroline reporter.

    PubMed

    Hu, Qiong; Zhou, Baojing; Dang, Pengyun; Li, Lianzhi; Kong, Jinming; Zhang, Xueji

    2017-01-15

    We report a versatile approach for the colorimetric assay of alkaline phosphatase (ALP) activity based on the distinctive metal-to-ligand charge-transfer (MLCT) absorption properties of Fe(II)-phenanthroline reporter. In the presence of ALP, the applied substrate ascorbic acid 2-phosphate is enzymatically hydrolyzed to produce ascorbic acid, which then reduces Fe(3+) to Fe(2+). The complexation of Fe(2+) with the bathophenanthroline disulfonate (BPS) ligand generates a blood-red Fe(BPS)3(4-) reporter, which is characterized by an intense MLCT absorption band at 535 nm in the visible range. Under optimal conditions, the spectral output exhibits a good quantitative relationship with ALP activity over the range of 0-220 mU mL(-1) with a detection limit of 0.94 mU mL(-1). Moreover, the activity of ALP can also be conveniently judged through naked-eye observations. Results indicate that it is highly selective and can be applied to the screening of ALP inhibitors. In addition, it has been successfully employed to detect the endogenous ALP level of undiluted human serum samples, with a detection limit of 1.05 mU mL(-1) being achieved. This approach avoids any elaborately designed substrates and holds considerable simplicity and flexibility for reporter design. This study broadens the horizon of the applications of phenanthroline-based transition metal complexes. Furthermore, an efficient and practical method like this has the potential to be widely used in clinical applications and in the point-of-care testing.

  6. Poly(vinyl chloride) tubing with covalently bound alkaline phosphatase and alternative approach for investigations of open-tubular bioreactors.

    PubMed

    Rozum, Beata; Gajownik, Kamil; Tymecki, Lukasz; Koncki, Robert

    2010-05-01

    A one-step carbodiimide method was found to allow covalent binding of enzymes to the inner wall of poly(vinyl chloride) (PVC) tubing. The immobilization is performed under mild conditions without laborious pretreatment or activation of reactor surface. In these preliminary studies, alkaline phosphatase (ALP, EC 3.1.3.1) and p-nitrophenyl phosphate (NPP) were applied as a model enzyme and substrate, respectively. The resulting open-tubular bioreactor exhibits satisfactory operational and storage stability. In addition, a novel and very simple instrumental concept for optical monitoring of the biocatalytic process directly inside the microbioreactor using a system of paired emitter-detector diodes is presented.

  7. Amplified thrombin aptasensor based on alkaline phosphatase and hemin/G-quadruplex-catalyzed oxidation of 1-naphthol.

    PubMed

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

    2015-05-20

    An alkaline phosphatase (ALP)-based biosensor can in situ generate an electroactive product by enzymatic hydrolysis of inactive substrates. To obtain a higher signal-to-background ratio, a chemical redox cycling signal-amplified strategy based on the addition of a strong reducing agent has often be applied in the construction of ALP-based biosensors. However, the strong reducing agent not only affects the activity of ALP but also readily reacts with dissolved oxygen, leading to inaccurate results. In this work, a new signal-amplified strategy for a thrombin (TB) aptasensor based on the catalytic oxidation of ALP-generated products, 1-naphthol (NP), using hemin/G-quadruplex DNAzymes was reported. We implemented gold-nanoparticle-decorated zinc oxide nanoflowers (Au-ZnO) as the matrix for immobilizing ALP and TB aptamer (TBA) and then labeled it with hemin to form hemin/G-quadruplex/ALP/Au-ZnO bioconjugates (TBA II bioconjugates). Through a "sandwich" reaction, TBA II bioconjugates were captured on the electrode surface. The amplified signal was carried out in two steps: (i) an ALP-catalyzed inactive substrate, 1-naphthyl phosphate (NPP), in situ produces NP on the surface of the electrode; (ii) on the one hand, NP as a new reactant could be directly electrooxidized and generated an electrochemical signal, but, on the other hand, NP could be oxidized by hemin/G-quadruplex in the presence of H2O2, resulting in amplification of the electrochemical signal. The proposed TB aptasensor achieved a linear range of 1 pM to 30 nM with a detection limit of 0.37 pM (defined as S/N = 3).

  8. Fluorescence Regulation of Poly(thymine)-Templated Copper Nanoparticles via an Enzyme-Triggered Reaction toward Sensitive and Selective Detection of Alkaline Phosphatase.

    PubMed

    Li, Junyao; Si, Ling; Bao, Jianchun; Wang, Zhaoyin; Dai, Zhihui

    2017-02-28

    The activity of alkaline phosphatase (ALP) is a crucial index of blood routine examinations, since the concentration of ALP is highly associated with various human diseases. To address the demands of clinical tests, efforts should be made to develop more approaches that can sense ALP in real samples. Recently, we find that fluorescence of poly(30T)-templated copper nanoparticles (CuNPs) can be directly and effectively quenched by pyrophosphate ion (PPi), providing new perspective in designing sensitive biosensors based on DNA-templated CuNPs. In addition, it has been confirmed that phosphate ion (Pi), product of PPi hydrolysis, does not affect the intense fluorescence of CuNPs. Since ALP can specifically hydrolyze PPi into Pi, fluorescence of CuNPs is thus regulated by an ALP-triggered reaction, and a novel ALP biosensor is successfully developed. As a result, ALP is sensitively and selectively quantified with a wide linear range of 6.0 × 10(-2) U/L to 6.0 × 10(2) U/L and a low detection limit of 3.5 × 10(-2) U/L. Besides, two typical inhibitors of ALP are evaluated by this analytical method, and different inhibitory effects are indicated. More importantly, by challenging this biosensor with real human serums, the obtained results get a fine match with the data from clinical tests, and the serum sample from a patient with liver disease is clearly distinguished, suggesting promising applications of this biosensor in clinical diagnosis.

  9. Detection of zeptomolar concentrations of alkaline phosphatase based on a tyrosinase and horse-radish peroxidase bienzyme biosensor.

    PubMed

    Ruan, C; Li, Y

    2001-07-06

    A bienzyme biosensor based on tyrosinase and horse-radish peroxidase is described in a flow injection analysis and cyclic voltammetry for measurement of phenol. Tyrosinase and horse-radish peroxidase were immobilized on the surface of a glassy carbon electrode by bovine serum albumin and glutaric dialdehyde. Phenol was oxidized by tyrosinase and horse-radish peroxidase via catechol to o-quinone in the presence of oxygen and hydrogen peroxide. The o-quinone was reduced to produce catechol (the substrate recycling) on the electrode surface. The enhanced sensitivity of the bienzyme electrode to phenol was observed in the flow injection system comparing with tyrosinase and horse-radish peroxidase monoenzyme electrodes. The mechanisms for enhanced amperometric response to phenol of bienzyme electrode were discussed. The biosensor was used to detect alkaline phosphatase (ALP). A detection limit of 1.4x10(-15) M ALP (140 zmol/100 mul) was obtained after 1 h incubation with phenyl phosphate.

  10. Investigation of the enzyme hydrolysis products of the substrates of alkaline phosphatase in electrochemical immunosensing.

    PubMed

    Preechaworapun, Anchana; Dai, Zong; Xiang, Yun; Chailapakul, Orawon; Wang, Joseph

    2008-07-15

    In this paper, we have critically evaluated the electrochemical behavior of the products of seven substrates of the enzyme label, alkaline phosphate, commonly used in electrochemical immunosensors. These products (and the corresponding substrates) include indigo carmine (3-indoyl phosphate), hydroquinone (hydroquinone diphosphate), 4-nitrophenol (4-nitrophenol phosphate), 4-aminophenol (p-aminophenyl phosphate), 1-naphthol (1-naphthyl phosphate), phenol (phenyl phosphate), and L-ascorbic acid (2-phospho-L-ascorbic acid). Cyclic voltammetry and amperometry of these products were carried out at glassy carbon (GC), screen-printed carbon (SPC) and gold (Au) electrodes, respectively. Among the products, L-ascorbic acid showed the most sensitive (24.8 microA cm(-2), 12.0 microA cm(-2), and 48.0 microA cm(-2) of 100 microM ascorbic acid at GC, SPC, and Au electrodes, respectively) and well-defined amperometric response at all electrodes used, making 2-phospho-l-ascorbic acid the best substrate in electrochemical detection involving an alkaline phosphatase (ALP) enzyme label. The 2-phospho-L-ascorbic acid is also commercially available and inexpensive. Therefore, it was the best choice for electrochemical detection using ALP as label. Using mouse IgG as a model, an ALP enzyme-amplified sandwich-type amperometric immunosensor was constructed. The immunosensor was designed by electropolymerization of o-aminobenzoic acid (o-ABA) conductive polymer on the surface of GC, SPC, and Au electrodes. The anti-mouse IgG was subsequently attached on the electrode surface through covalent bonding between IgG antibody and the carboxyl groups from poly(o-ABA). Using 2-phospho-L-ascorbic acid as a substrate, the poly(o-ABA)/Au immunosensor produced the best signal (about 297 times of current density response ratio between 1000 ng mL(-1) and 0 ng mL(-1) of mouse IgG), demonstrating that amperometric immunosensors based on a conducting polymer electrode system were sensitive to

  11. PAPSS2 Promotes Alkaline Phosphates Activity and Mineralization of Osteoblastic MC3T3-E1 Cells by Crosstalk and Smads Signal Pathways

    PubMed Central

    Wang, Weizhuo; Li, Fang; Wang, Kunzheng; Cheng, Bin; Guo, Xiong

    2012-01-01

    Several studies have indicated that PAPSS2 (3′-phosphoadenosine-5′-phosphosulfate synthetase 2) activity is important to normal skeletal development. Mouse PAPSS2 is predominantly expressed during the formation of the skeleton and cartilaginous elements of the mouse embryo and in newborn mice. However, the role and mechanism of PAPSS2 in bone formation remains largely unidentified. By analyzing the expression pattern of the PAPSS2 gene, we have found that PAPSS2 is expressed in bone tissue and bone formation. PAPSS2 transcripts increase during osteoblast differentiation and are in less level in RANKL-induced osteoclast like cells. By using lentivirus-mediated RNA interference (RNAi) technology, we knocked down PAPSS2 expression in MC3T3-E1 osteoblast. Silencing of PAPSS2 expression significantly decreases ALP activity and cell mineralization, inhibits expression of osteoblast marker osteopontin (OPN) and collagen I. Conversely, overexpression of PAPSS2 promotes the MC3T3-E1 to differentiate into osteoblast and mineralization. Moreover, compared to that in the control cells, the mRNA level and protein expression of phosphorylated Smad 2/3, which is a key transcriptional factor in the Smad osteoblast differentiation pathway, showed significant decreases in PAPSS2-silenced cells and increases in PAPSS2-overexpression cells. These results suggest that PAPSS2 might regulate osteoblast ALP activity and cell mineralization, probably through Smads signal pathways. PMID:22916269

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

    PubMed

    Obayashi, Yusuke; Iino, Ryota; Noji, Hiroyuki

    2015-08-07

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

  13. Simultaneous demonstration of bone alkaline and acid phosphatase activities in plastic-embedded sections and differential inhibition of the activities.

    PubMed

    Liu, C; Sanghvi, R; Burnell, J M; Howard, G A

    1987-01-01

    Bone alkaline (AlP) and acid phosphatase (AcP) activities were simultaneously demonstrated in tissue sections obtained from mice, rats, and humans. The method involved tissue fixation in ethanol, embedding in glycol methacrylate (GMA), and demonstration of AlP and AcP activities employing a simultaneous coupling azo dye technique using substituted naphthol phosphate as a substrate. AlP activity was demonstrated first followed by AcP activity. Both enzyme activities were demonstrated in tissue sections from bones fixed and/or stored in acetone or 70% ethanol for up to 14 days or stored in GMA for 2 months. AlP activity in tissue sections from bones fixed in 10% formalin, 2% glutaraldehyde, or formal-calcium, however, was markedly inhibited after 3-7 days and was no longer detectable after 14 days of fixation. Moreover, AlP activity was diminished in tissue sections from bones fixed in 70% ethanol or 10% formalin and subsequently demineralized in 10% EDTA (pH 7) for 2 days, and the activity was completely abolished in tissue sections from bones subsequently demineralized in 5% formic acid: 20% sodium citrate (1:1, pH 4.2) for 2 days. Methyl methacrylate (MMA) embedding at concentrations above 66% completely inhibited AlP activity. AcP activity, however, was only partially inhibited by formalin, glutaraldehyde, or formal-calcium after 7 or 14 days of fixation or by MMA embedding and was unaffected by the demineralizing agent formic acid-citrate for 2 days. While AcP activity was preserved in bones fixed in formalin and subsequently demineralized in EDTA, the activity was completely abolished when EDTA demineralization was carried out on bones previously fixed in 70% ethanol.(ABSTRACT TRUNCATED AT 250 WORDS)

  14. 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.

  15. QM/MM analysis suggests that Alkaline Phosphatase (AP) and Nucleotide pyrophosphatase/phosphodiesterase slightly tighten the transition state for phosphate diester hydrolysis relative to solution: implication for catalytic promiscuity in the AP superfamily

    PubMed Central

    Hou, Guanhua

    2011-01-01

    Several members of the Alkaline Phosphatase (AP) superfamily exhibit a high level of catalytic proficiency and promiscuity in structurally similar active sites. A thorough characterization of the nature of transition state for different substrates in these enzymes is crucial for understanding the molecular mechanisms that govern those remarkable catalytic properties. In this work, we study the hydrolysis of a phosphate diester, MpNPP−, in solution, two experimentally well-characterized variants of AP (R166S AP, R166S/E322Y AP) and wild type Nucleotide pyrophosphatase/phosphodiesterase (NPP) by QM/MM calculations in which the QM method is an approximate density functional theory previously parameterized for phosphate hydrolysis (SCC-DFTBPR). The general agreements found between these calculations and available experimental data for both solution and enzymes support the use of SCC-DFTBPR/MM for a semi-quantitative analysis of the catalytic mechanism and nature of transition state in AP and NPP. Although phosphate diesters are cognate substrates for NPP but promiscuous substrates for AP, the calculations suggest that their hydrolysis reactions catalyzed by AP and NPP feature similar synchronous transition states that are slightly tighter in nature compared to that in solution, due in part to the geometry of the bimetallic zinc motif. Therefore, this study provides the first direct computational support to the hypothesis that enzymes in the AP superfamily catalyze cognate and promiscuous substrates via similar transition states to those in solution. Our calculations do not support the finding of recent QM/MM studies by López-Canut and coworkers, who suggested that the same diester substrate goes through a much looser transition state in NPP/AP than in solution, a result likely biased by the large structural distortion of the bimetallic zinc site in their simulations. Finally, our calculations for different phosphate diester orientations and phosphorothioate diesters

  16. QM/MM analysis suggests that Alkaline Phosphatase (AP) and nucleotide pyrophosphatase/phosphodiesterase slightly tighten the transition state for phosphate diester hydrolysis relative to solution: implication for catalytic promiscuity in the AP superfamily.

    PubMed

    Hou, Guanhua; Cui, Qiang

    2012-01-11

    Several members of the Alkaline Phosphatase (AP) superfamily exhibit a high level of catalytic proffciency and promiscuity in structurally similar active sites. A thorough characterization of the nature of transition state for different substrates in these enzymes is crucial for understanding the molecular mechanisms that govern those remarkable catalytic properties. In this work, we study the hydrolysis of a phosphate diester, MpNPP(-), in solution, two experimentally well-characterized variants of AP (R166S AP, R166S/E322Y AP) and wild type Nucleotide pyrophosphatase/phosphodiesterase (NPP) by QM/MM calculations in which the QM method is an approximate density functional theory previously parametrized for phosphate hydrolysis (SCC-DFTBPR). The general agreements found between these calculations and available experimental data for both solution and enzymes support the use of SCC-DFTBPR/MM for a semiquantitative analysis of the catalytic mechanism and nature of transition state in AP and NPP. Although phosphate diesters are cognate substrates for NPP but promiscuous substrates for AP, the calculations suggest that their hydrolysis reactions catalyzed by AP and NPP feature similar synchronous transition states that are slightly tighter in nature compared to that in solution, due in part to the geometry of the bimetallic zinc motif. Therefore, this study provides the first direct computational support to the hypothesis that enzymes in the AP superfamily catalyze cognate and promiscuous substrates via similar transition states to those in solution. Our calculations do not support the finding of recent QM/MM studies by López-Canut and co-workers, who suggested that the same diester substrate goes through a much looser transition state in NPP/AP than in solution, a result likely biased by the large structural distortion of the bimetallic zinc site in their simulations. Finally, our calculations for different phosphate diester orientations and phosphorothioate diesters

  17. 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

  18. Fabrication of calcium phosphate films for coating on titanium substrates heated up to 773 K by RF magnetron sputtering and their evaluations.

    PubMed

    Ueda, Kyosuke; Narushima, Takayuki; Goto, Takashi; Taira, Masayuki; Katsube, Tomoyuki

    2007-09-01

    Calcium phosphate films were fabricated on titanium substrates heated up to 773 K using radiofrequency (RF) magnetron sputtering. The deposition rate, phase and preferred orientation of the calcium phosphate films were studied. Immersion tests for the films were conducted using Hanks' solution and PBS(-), and the surface reactions on the specimens coated with the calcium phosphate films were investigated. The bonding strength between the coating films and the titanium substrates before and after the immersion tests was evaluated; the bonding strength decreased after the immersion tests. The alkaline phosphatase (ALP) activity of SaOS-2 cells on a titanium plate coated with a calcium phosphate film was examined by conducting a culture test. Calcium phosphate coating increased the ALP activity of SaOS-2 cells cultured for 3 and 7 days. Titanium cylinders were coated with an amorphous calcium phosphate film and implanted into the mandibles of beagle dogs. An increase in the extent of bone-implant contact for the coated titanium cylinders was confirmed 8 to 12 weeks after implantation and compared with the case for uncoated titanium cylinders.

  19. Biological performance of titania containing phosphate-based glasses for bone tissue engineering applications.

    PubMed

    Abou Neel, Ensanya Ali; Chrzanowski, Wojciech; Knowles, Jonathan Campbell

    2014-02-01

    The interplay between glass chemistry, structure, degradation kinetics, and biological activity provides flexibility for the development of scaffolds with highly specific cellular response. The aim of this study was therefore to investigate the role of titania inclusion into the phosphate-based glass on its ability to stimulate osteoblast-like human osteosarcoma (HOS) cells to adhere, proliferate and differentiate. In depth morphological and biochemical characterisation was performed on HOS cells cultured on the surface of glass discs. Cell proliferation was also studied in the presence of the glass extract. Cell differentiation, through osteoblast phenotype genes, alkaline phosphatase (ALP) activity and osteocalcin production, was carried out using normal or osteogenic media. Both Thermanox® and titania free glass were used as controls. The data demonstrated that titania inclusion provides desired cytocompatible surface that supported initial cell attachment, sustained viability, and increased cell proliferation similar or significantly higher than Thermanox®. The modified glasses regulated osteoblastic cell differentiation as detected by osteoblast phenotype gene transcription and upregulated ALP and osteocalcin expression. Using osteogenic media had no significant effect on ALP activity and osteocalcin expression. Therefore, titania modified phosphate glasses may have future use as bone tissue engineering scaffolds.

  20. A rapid, quantitative assay for measuring alkaline phosphatase activity in osteoblastic cells in vitro.

    PubMed

    Sabokbar, A; Millett, P J; Myer, B; Rushton, N

    1994-10-01

    Alkaline phosphatase (ALP) is the most widely recognized biochemical marker for osteoblast activity. Although its precise function is poorly understood, it is believed to play a role in skeletal mineralization. The aim of this study was to develop an assay suitable for measuring the activity of this enzyme in microtiter plate format. Using the well-characterized osteoblast-like cell line Saos-2, this paper describes an optimized biochemical assay suitable for measuring ALP activity in tissue culture samples. We have determined that a p-nitrophenyl phosphate substrate concentration of 9 mM provides highest enzyme activities. We have found that cell concentration, and hence enzyme concentration, affects both the kinetics and precision of the assay. We also tested several methods of enzyme solubilization and found that freeze-thawing the membrane fractions twice at -70 degrees C/37 degrees C or freeze-thawing once with sonication yielded highest enzyme activities. The activity of the enzyme decreased by 10% after 7 days storage. This assay provides a sensitive and reproducible method that is ideally suited for measuring ALP activity in isolated osteoblastic cells, although sample preparation and storage can influence results.

  1. ALP isoenzyme test

    MedlinePlus

    Alkaline phosphatase isoenzyme test ... anything for 10 to 12 hours before the test, unless your health care provider tells you to do so. Many medicines can interfere with blood test results. Your health care provider will tell you ...

  2. 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.

  3. Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO2/g-C3N4, PAMAM Dendrimer, and Alkaline Phosphatase.

    PubMed

    Li, Xue; Zhu, Lusheng; Zhou, Yunlei; Yin, Huanshun; Ai, Shiyun

    2017-02-21

    A novel photoelectrochemical (PEC) assay is developed for sensitive detection of protein kinase A (PKA) activity based on PKA-catalyzed phosphorylation reaction in solution and signal amplification strategy triggered by PAMAM dendrimer and alkaline phosphatase (ALP). In this strategy, it is noteworthy at this point that PKA phosphorylation was achieved in solution instead of on the surface of the electrode, which has advantages of the good contact in reactants and simple experimental procedure. For immobilizing the phosphorylated peptide (P-peptide) on electrode surface, graphite-like carbon nitride (g-C3N4) and titanium dioxide (TiO2) complex is synthesized and characterized, which plays a significant role for TiO2 conjugating phosphate groups and g-C3N4 providing PEC signal. Subsequently, PAMAM dendrimer and ALP can be captured on P-peptide and TiO2/g-C3N4 modified ITO electrode via interaction between the -COOH groups on the surface of PAMAM dendrimer and the -NH2 groups of peptide and ALP, which can lead to the increase of ALP amount on the modified electrode surface assisted with the PAMAM dendrimer. As a result, the amount of ALP catalyzes of L-ascorbic acid 2-phosphate trisodium salt (AAP) to produce electron donor of ascorbic acid (AA), resulting in an increased photocurrent. The proposed detection assay displays high selectivity and low detection limit of 0.048 U/mL (S/N = 3) for PKA activity. This biosensor can also be applied for the evaluation of PKA inhibition and PKA activity assay in cell samples. Therefore, the fabricated PEC biosensor is potentionally well in PKA activity detection and inhibitor screening.

  4. Fibronectin-calcium phosphate composite layer on hydroxyapatite to enhance adhesion, cell spread and osteogenic differentiation of human mesenchymal stem cells in vitro.

    PubMed

    Sogo, Yu; Ito, Atsuo; Matsuno, Tomonori; Oyane, Ayako; Tamazawa, Gaku; Satoh, Tazuko; Yamazaki, Atsushi; Uchimura, Eiji; Ohno, Tadao

    2007-06-01

    Fibronectin (Fn) and type I collagen (Col) were immobilized on a surface of a hydroxyapatite (HAP) ceramic by coprecipitation with calcium phosphate in a supersaturated calcium phosphate solution prepared by mixing clinically approved infusion fluids. These proteins and the calcium phosphate precipitate formed a composite surface layer. As a result, the proteins were immobilized firmly as not to be released completely for 3 d in a physiological salt solution. When human mesenchymal stem cells (hMSCs) were cultured on a HAP ceramic in a differentiation medium supplemented with dexamethasone, beta-glycerophosphate and ascorbic acid, hMSCs spread well within 1 h. The alkaline phosphatase (ALP) activity of hMSCs cultured on the Fn-calcium phosphate composite layer significantly increased compared with that of hMSCs cultured on the untreated HAP ceramic. On the other hand, Col did not increase the ALP activity of hMSCs and no synergy between Fn and Col was observed. Therefore, the Fn-calcium phosphate composite layer formed on the HAP is useful for the enhancement of the spreading and osteogenic differentiation of hMSCs in vitro.

  5. 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

  6. Enzymatic mineralization of hydrogels for bone tissue engineering by incorporation of alkaline phosphatase.

    PubMed

    Douglas, Timothy E L; Messersmith, Philip B; Chasan, Safak; Mikos, Antonios G; de Mulder, Eric L W; Dickson, Glenn; Schaubroeck, David; Balcaen, Lieve; Vanhaecke, Frank; Dubruel, Peter; Jansen, John A; Leeuwenburgh, Sander C G

    2012-08-01

    Alkaline phosphatase (ALP), an enzyme involved in mineralization of bone, is incorporated into three hydrogel biomaterials to induce their mineralization with calcium phosphate (CaP). These are collagen type I, a mussel-protein-inspired adhesive consisting of PEG substituted with catechol groups, cPEG, and the PEG/fumaric acid copolymer OPF. After incubation in Ca-GP solution, FTIR, EDS, SEM, XRD, SAED, ICP-OES, and von Kossa staining confirm CaP formation. The amount of mineral formed decreases in the order cPEG > collagen > OPF. The mineral:polymer ratio decreases in the order collagen > cPEG > OPF. Mineralization increases Young's modulus, most profoundly for cPEG. Such enzymatically mineralized hydrogel/CaP composites may find application as bone regeneration materials.

  7. Enzymatic mineralization of hydrogels for bone tissue engineering by incorporation of alkaline phosphatase

    PubMed Central

    Douglas, Timothy E.L.; Messersmith, Philip B.; Chasan, Safak; Mikos, Antonios G.; de Mulder, Eric L.W.; Dickson, Glenn; Schaubroeck, David; Balcaen, Lieve; Vanhaecke, Frank; Dubruel, Peter; Jansen, John A.

    2013-01-01

    Alkaline Phosphatase (ALP), an enzyme involved in mineralization of bone, was incorporated into three hydrogel biomaterials to induce their mineralization with calcium phosphate (CaP). These were collagen type I, a mussel protein-inspired adhesive consisting of PEG substituted with catechol groups, cPEG, and the PEG-fumaric acid copolymer OPF. After incubation in calcium glycerophosphate (Ca-GP) solution, FTIR, EDS, SEM, XRD, SAED, ICP-OES and von Kossa staining confirmed CaP formation. The amount of mineral formed decreased in the order cPEG > collagen > OPF. Mineral:polymer ratio decreased in the order collagen > cPEG > OPF. Mineralization increased Young’s modulus, most profoundly for cPEG. Such enzymatically mineralized hydrogel-CaP composites could find application as bone regeneration materials. PMID:22648976

  8. 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.

  9. 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

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

    PubMed

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

    2016-03-05

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

  12. 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.

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

    PubMed

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

    2016-09-15

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

  14. On wet chemical phosphorus recovery from sewage sludge ash by acidic or alkaline leaching and an optimized combination of both.

    PubMed

    Petzet, Sebastian; Peplinski, Burkhard; Cornel, Peter

    2012-08-01

    The advantages and drawbacks of existing wet chemical phosphorus (P) recovery technologies, their applicability to different types of sewage sludge ash (SSA) and the role of the decay products of detergent zeolites as a source of reactive Al in SSA are analyzed. Since neither a purely acidic nor a purely alkaline treatment are able to provide satisfactory technical solutions a wet chemical phosphorus (P) recovery process for sewage sludge ashes (SSAs) is investigated in detail that is based on a sequential treatment of SSA with an acid and a base. As a result of an acidic pre-treatment, the P fraction of the raw SSA that was bound as - alkaline-insoluble - calcium phosphate (Ca-P) is converted into aluminum phosphate (Al-P). This newly formed Al-P can be easily dissolved via alkaline treatment and then easily separated from the alkaline leachate via precipitation of Ca-P. The Al-component can be reused as precipitant for P-removal in waste water treatment plants (WWTPs). The investigated process requires fewer chemicals than the direct acidic dissolution of all P-compounds contained in the SSA. This is due to the described rearrangement of the P component from Ca-P to Al-P. That such a rearrangement of P occurs indeed was confirmed through a combination of XRD, ICP and XRF analyses together with mass balance calculations. The present investigation proves that the process works for very different types of SSAs: For Al-rich SSAs that come from WWTPs where Al-salt is used for chemical P-removal the described sequential treatment process works best and yields P-recovery rates as high as 70-77%. But even for SSAs from WWTPs where only iron salt is used for chemical P-removal, a considerable amount of the reactive Al necessary for the described P-rearrangement is supplied by decay products of detergent zeolites, a hidden Al-source present in most SSAs produced in Europe.

  15. Genetic responses to nanostructured calcium-phosphate-coated implants.

    PubMed

    Jimbo, R; Xue, Y; Hayashi, M; Schwartz-Filho, H O; Andersson, M; Mustafa, K; Wennerberg, A

    2011-12-01

    Nanostructured calcium phosphate (CaP) has been histologically and biomechanically proven to enhance osseointegration of implants; however, conventional techniques were not sufficiently sensitive to capture its biological effects fully. Here, we compared the conventional removal torque (RTQ) evaluation and gene expression in tissues around nanostructured CaP-coated implants, using real-time RT-PCR, with those of uncoated implants, in a rabbit model. At 2 wks, RTQ values were significantly higher, alkaline phosphatase (ALP) expression was significantly higher, and runt-related transcription factor 2 and tumor necrosis factor-α expressions were significantly lower in the coated than in the uncoated implants. This indicates that inflammatory responses were suppressed and osteoprogenitor activity increased around the CaP-coated surface. At 4 wks, although RTQ values did not significantly differ between the 2 groups, ALP and osteocalcin (OCN) were significantly up-regulated in the coated group, indicating progressive mineralization of the bone around the implant. Moreover, an osteoclast marker, adenosine triphosphatase, which indicates acidification of the resorption lacunae, was significantly higher for the coated implants, suggesting gradual resorption of the CaP coating. This study reveals detailed genetic responses to nanostructured CaP-coated implants and provides evidence that the effect of nanotopography is significant during the osseointegration cascade.

  16. ALPS: Advanced Learning Packages, 1978-1979.

    ERIC Educational Resources Information Center

    San Juan Unified School District, Carmichael, CA.

    The document describes the ALPS (Advanced Learning Packages) program for teaching gifted students. Introductory materials provide information on teacher requirements, school requirements, ALPS teacher orientation responsibilities, orientation week, field trip procedures, gifted money available, ALPS costs, ALPS evaluations, the Structure of…

  17. 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.

  18. Adult Learning Program Service (ALPS).

    ERIC Educational Resources Information Center

    Carlisle, Robert

    The Adult Learning Program Service (ALPS) aims to reach eight and a half million adults between ages 25 and 44 and teach them reading and math skills they can use at home and on the job. ALPS proposes to reach those who have never finished high school but do have at least a sixth-grade reading level. They could use their new skills to prepare for…

  19. Bone cell responses to the composite of Ricinus communis polyurethane and alkaline phosphatase.

    PubMed

    Beloti, Marcio Mateus; de Oliveira, Paulo Tambasco; Tagliani, Marcela Martini; Rosa, Adalberto Luiz

    2008-02-01

    The aim of this study was to evaluate the response of osteoblastic cells to the composite of Ricinus communis polyurethane (RCP) and alkaline phosphatase (ALP) incubated in synthetic body fluid (SBF). RCP pure (RCPp) and RCP blended with ALP 6 mg/mL polymer (RCP+ALP) were incubated in SBF for 17 days. Four groups of RCP were tested: RCPp, RCP+ALP, and RCPp and RCP+ALP incubated in SBF (RCPp/SBF and RCP+ALP/SBF). Stem cells from rat bone marrow were cultured in conditions that allowed osteoblastic differentiation on RCP discs and were evaluated: cell adhesion, culture growth, cell viability, total protein content, ALP activity, and bone-like nodule formation. Data were compared by ANOVA or Kruskal-Wallis test. The group RCP+ALP was highly cytotoxic and, therefore, was not considered here. Cell adhesion (p = 0.14), culture growth (p = 0.39), viability (p = 0.46) and total protein content (p = 0.12) were not affected by either RCP composition or incubation in SBF. ALP activity was affected (p = 0.0001) as follows: RCPp < RCPp/SBF < RCP+ALP/SBF. Bone-like nodule formation was not observed on all evaluated groups. The composite RCP+ALP prior to SBF incubation is cytotoxic and must not be considered as biomaterial, but the incorporation of ALP to the RCP followed by SBF incubation could be a useful alternative to improve the biological properties of the RCP.

  20. Phosphate removal from domestic wastewater using thermally modified steel slag.

    PubMed

    Yu, Jian; Liang, Wenyan; Wang, Li; Li, Feizhen; Zou, Yuanlong; Wang, Haidong

    2015-05-01

    This study was performed to investigate the removal of phosphate from domestic wastewater using a modified steel slag as the adsorbent. The adsorption effects of alkalinity, salt, water, and thermal modification were investigated. The results showed that thermal activation at 800°C for 1 hr was the optimum operation to improve the adsorption capacity. The adsorption process of the thermally modified slag was well described by the Elovich kinetic model and the Langmuir isotherm model. The maximum adsorption capacity calculated from the Langmuir model reached 13.62 mg/g. Scanning electron microscopy indicated that the surface of the modified slag was cracked and that the texture became loose after heating. The surface area and pore volume did not change after thermal modification. In the treatment of domestic wastewater, the modified slag bed (35.5 kg) removed phosphate effectively and operated for 158 days until the effluent P rose above the limit concentration of 0.5 mg/L. The phosphate fractionation method, which is often applied in soil research, was used to analyze the phosphate adsorption behavior in the slag bed. The analysis revealed that the total contents of various Ca-P forms accounted for 81.4%-91.1%, i.e., Ca10-P 50.6%-65.1%, Ca8-P 17.8%-25.0%, and Ca2-P 4.66%-9.20%. The forms of Al-P, Fe-P, and O-P accounted for only 8.9%-18.6%. The formation of Ca10-P precipitates was considered to be the main mechanism of phosphate removal in the thermally modified slag bed.

  1. 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.

  2. Small interfering RNA of alkaline phosphatase inhibits matrix mineralization.

    PubMed

    Kotobuki, Noriko; Matsushima, Asako; Kato, Youichi; Kubo, Yoko; Hirose, Motohiro; Ohgushi, Hajime

    2008-05-01

    To investigate the cascade of matrix mineralization, cells expressing high and low alkaline phosphatase (ALP) were separated from human osteoblast-like (HOS) cells by fluorescence-activated cell sorting with an ALP antibody. After these cells had been recloned from single cells and then cultured under osteogenic conditions, high-ALP-expressing HOS (H-HOS) cells showed matrix mineralization, but low-ALP-expressing HOS (L-HOS) cells did not. The interaction among osteogenic-related genes, such as runt-related transcription factor 2 (RUNX2), collagen type I alpha1 chain (COL1A1), tissue non-specific ALP, and osteocalcin (OCN), is well known as being related to matrix mineralization. Quantitative real-time polymerase chain reaction revealed that the gene expression of ALP was higher in H-HOS cells than in L-HOS, whereas the gene expression of RUNX2, COL1A1, and OCN was lower in H-HOS cells than in L-HOS cells. When small interfering RNAs (siRNAs) of these osteogenic-related genes were introduced into H-HOS cells by transfection, only ALP siRNA inhibited matrix mineralization. Furthermore, the expression of not only the ALP gene, but also the COL1A1 and RUNX2 genes was influenced by the inhibition of ALP, although the expression of OCN was not affected by the inhibition of ALP. We have been able to confirm that the ALP gene is a strong candidate as the trigger of matrix mineralization. These results indicate the usefulness of cloned osteogenic cells in investigating the molecular mechanisms of matrix mineralization, the function of which can be modulated by using a variety of siRNAs.

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

    PubMed

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

    2011-10-01

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

  4. 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.

  5. Osteogenic differentiation of osteoblasts induced by calcium silicate and calcium silicate/β-tricalcium phosphate composite bioceramics.

    PubMed

    Fei, Lisha; Wang, Chen; Xue, Yang; Lin, Kaili; Chang, Jiang; Sun, Jiao

    2012-07-01

    In this study, calcium silicate (CS) and CS/β-tricalcium phosphate (CS/β-TCP) composites were investigated on their mechanism of osteogenic proliferation and differentiation through regulating osteogenic-related gene and proteins. Osteoblast-like cells were cultured in the extracts of these CS-based bioceramics and pure β-TCP, respectively. The main ionic content in extracts was analyzed by inductively coupled plasma-atomic emission spectroscopy. The cell viability, mineralization, and differentiation were evaluated by MTT assay, Alizarin Red-S staining and alkaline phosphatase (ALP) activity assay. The expressions of BMP-2, transforming growth factor-β (TGF-β), Runx2, ALP, and osteocalcin (OCN) at both gene and protein level were detected by real-time polymerase chain reaction analysis and Western blot. The result showed that the extracts of CS-based bioceramics promoted cells proliferation, differentiation, and mineralization when compared with pure β-TCP. Accordingly, pure CS and CS/β-TCP composites stimulated osteoblast-like cells to express BMP-2/TGF-β gene and proteins, and further regulate the expression of Runx2 gene and protein, and ultimately affect the ALP activity and OCN deposition. This study suggested that the CS-based bioceramics could not only promote the expression of osteogenic-related genes but also enhance the genes to encode the corresponding proteins, which could finally control osteoblast-like cells proliferation and differentiation.

  6. The mechanical and biological properties of an injectable calcium phosphate cement-fibrin glue composite for bone regeneration.

    PubMed

    Cui, Geng; Li, Jie; Lei, Wei; Bi, Long; Tang, Peifu; Liang, Yutian; Tao, Sheng; Wang, Yan

    2010-02-01

    Calcium phosphate cement (CPC) that can be injected to form a scaffold in situ has promise for the repair of bone defects. However, its low-strength limits the CPC to non-stress-bearing repairs. Fibrin glue (FG) with good sticking property and biocompatibility is possible used to reinforce the CPC. The objective of this study was to investigate the effects of FG on the mechanical and biological properties of CPC in an injectable CPC-FG composite. The initial setting time of this CPC-FG was delayed compared with the CPC control at different powder/liquid (P/L) mass ratio (p > 0.05). At a P/L of 5, the strength was (38.41 +/- 4.32) MPa for the CPC-FG, much higher than (27.42 +/- 2.85) MPa for the CPC alone (p < 0.05). SEM showed bone marrow stromal cells (BMSCs) with healthy spreading and anchored on the CPC-FG composite. After 14 days, the alkaline phosphatase (ALP) activity was (538 +/- 33) for the BMSCs on the CPC-FG and (517 +/- 27) for the BMSCs on the CPC alone. Both ALPs were higher than the baseline ALP (93 +/- 10) for the undifferentiated BMSCs (p < 0.05). The results demonstrate that this stronger CPC-FG scaffold may be useful for stem cell-based bone regeneration in moderate load-bearing orthopedic applications.

  7. Isozyme profile and tissue-origin of alkaline phosphatases in mouse serum

    PubMed Central

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

    2013-01-01

    Mouse serum alkaline phosphatase (ALP) is frequently measured and interpreted in mammalian bone research, however; little is known about the circulating ALPs in mice and their relation to human ALP isozymes and isoforms. Mouse ALP was extracted from liver, kidney, intestine, and bone from vertebra, femur and calvaria tissues. Serum from mixed strains of wild-type (WT) mice and from individual ALP knockout strains were investigated, i.e., Alpl−/− (a.k.a. Akp2 encoding tissue-nonspecific ALP or TNALP), Akp3−/− (encoding duodenum-specific intestinal ALP or dIALP), and Alpi−/− (a.k.a. Akp6 encoding global intestinal ALP or gIALP). The ALP isozymes and isoforms were identified by various techniques and quantified by high-performance liquid chromatography. Results from the WT and knockout mouse models revealed identical bone-specific ALP isoforms (B/I, B1, and B2) as found in human serum, but in addition mouse serum contains the B1x isoform only detected earlier in patients with chronic kidney disease and in human bone tissue. The two murine intestinal isozymes, dIALP and gIALP, were also identified in mouse serum. All four bone-specific ALP isoforms (B/I, B1x, B1, and B2) were identified in bone tissues from mice, in good correspondence with those found in human bones. All mouse tissues, except liver and colon, contained significant ALP activities. This is a notable difference as human liver contains vast amounts of ALP. Histochemical staining, Northern and Western blot analysis confirmed undetectable ALP expression in liver tissue. ALP activity staining showed some positive staining in the bile canaliculi for BALB/c and FVB/N WT mice, but not in C57Bl/6 and ICR mice. Taken together, while the main source of ALP in human serum originates from bone and liver, and a small fraction from intestine (<5%), mouse serum consists mostly of bone ALP, including all four isoforms, B/I, B1x, B1, and B2, and two intestinal ALP isozymes dIALP and gIALP. We suggest that the

  8. Significantly Elevated Liver Alkaline Phosphatase in Congestive Heart Failure

    PubMed Central

    Shamban, Leonid; Patel, Brijesh; Williams, Michael

    2014-01-01

    Congestive hepatopathy can have a mildly elevated liver profile, which should normalize with appropriate therapy. Liver specific alkaline phosphatase (ALP) in decompensated heart failure (HF) can be mildly elevated. The levels exceeding beyond the expected rise should be a concern and lead to further investigation. The literature reports insubstantial number of cases regarding significantly elevated levels of ALP and congestive hepatopathy. We report a case of a 45-year-old female with known history of severe cardiomyopathy that had persistently elevated levels of ALP. The extensive workup was negative for any specific pathology. The liver biopsy was consistent with congestive hepatopathy. The patient’s ALP levels decreased with aggressive diuretic therapy but still remained elevated. PMID:27785272

  9. Canine serum thermostable alkaline phosphatase isoenzyme from a dog with hepatocellular carcinoma.

    PubMed

    Fukui, Yu-ichi; Sato, Jun; Sato, Reeko; Yasuda, Jun; Naito, Yoshihisa

    2006-10-01

    A dog histopathologically diagnosed with hepatocellular carcinoma (HCC) showed very high serum alkaline phosphatase (ALP) activity. A supernatant of ascitic fluid and tumor tissue extracted from the dog also showed much higher ALP activity than normal. ALP isoenzyme analysis of samples was performed using polyacrylamide gel disk electrophoresis, and a wide, broad abnormal band was observed. By various treatments, the abnormal band showed thermostability, which is a characteristic of tumor-associated ALP that has only been reported in humans. The thermostable ALP isoenzyme was not found in sera from 39 dogs with several types of tumor that originated from the liver, except for HCC, nor was it found in 10 dogs with hepatic diseases that did not include hepatic tumors. The thermostable ALP isoenzyme seemed to be associated with canine HCC.

  10. Proteomic responses of oceanic Synechococcus WH8102 to phosphate and zinc scarcity and cadmium additions

    PubMed Central

    Cox, Alysia D.; Saito, Mak A.

    2013-01-01

    Synechococcus sp. WH 8102 is a motile marine cyanobacterium isolated originally from the Sargasso Sea. To test the response of this organism to cadmium (Cd), generally considered a toxin, cultures were grown in a matrix of high and low zinc (Zn) and phosphate (PO43−) and were then exposed to an addition of 4.4 pM free Cd2+ at mid-log phase and harvested after 24 h. Whereas Zn and PO43− had little effect on overall growth rates, in the final 24 h of the experiment three growth effects were noticed: (i) low PO43− treatments showed increased growth rates relative to high PO43− treatments, (ii) the Zn/high PO43− treatment appeared to enter stationary phase, and (iii) Cd increased growth rates further in both the low PO43− and Zn treatments. Global proteomic analysis revealed that: (i) Zn appeared to be critical to the PO43− response in this organism, (ii) bacterial metallothionein (SmtA) appears correlated with PO43− stress-associated proteins, (iii) Cd has the greatest influence on the proteome at low PO43− and Zn, (iv) Zn buffered the effects of Cd, and (v) in the presence of both replete PO43− and added Cd the proteome showed little response to the presence of Zn. Similar trends in alkaline phosphate (ALP) and SmtA suggest the possibility of a Zn supply system to provide Zn to ALP that involves SmtA. In addition, proteome results were consistent with a previous transcriptome study of PO43− stress (with replete Zn) in this organism, including the greater relative abundance of ALP (PhoA), ABC phosphate binding protein (PstS) and other proteins. Yet with no Zn in this proteome experiment the PO43− response was quite different including the greater relative abundance of five hypothetical proteins with no increase in PhoA or PstS, suggesting that Zn nutritional levels are connected to the PO43− response in this cyanobacterium. Alternate ALP PhoX (Ca) was found to be a low abundance protein, suggesting that PhoA (Zn, Mg) may be more

  11. Configurational effects of collagen/ALP coatings on enzyme immobilization and surface mineralization

    NASA Astrophysics Data System (ADS)

    Bosco, R.; Leeuwenburgh, S. C. G.; Jansen, J. A.; van den Beucken, J. J. J. P.

    2014-08-01

    The ultimate goal for surface modifications in bone implants is to achieve biologically active surface able to control and trigger specific tissue response. In this study was evaluated the effects of organic compound, derived from extracellular matrix, involved in tissue mineralization. Alkaline phosphatase (ALP) plays a fundamental role in bone mineralization concurrently with collagen, the main organic components of bones. Electrospray deposition (ESD) was used to coat titanium disks with ALP and collagen at room temperature. To verify the synergistic role of ALP and collagen different conformations of coatings (mixed and layered) were obtained and their mineralization capacity was tested in vitro. The mineralization tests indicated the fundamental role of collagen to increase ALP coating retention. Analyses indicated that the coating conformation has a role; in fact the mixed group showed improved ALP retention, enzymatic activity and unique mineralized surface morphology. ESD demonstrated to be a successful method to deposit organic molecules preserving their properties as indicated by the in vitro results. These findings proved the synergistic effect of ALP and collagen in inducing mineralization offering an intriguing coating constituent for medical device that aim to trigger surface mineralization such as bone implants.

  12. 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.

  13. Matrix gla protein and alkaline phosphatase are differently modulated in human dermal fibroblasts from PXE patients and controls.

    PubMed

    Boraldi, Federica; Annovi, Giulia; Vermeer, Cees; Schurgers, Leon J; Trenti, Tommaso; Tiozzo, Roberta; Guerra, Deanna; Quaglino, Daniela

    2013-04-01

    Mineralization of elastic fibers in pseudoxanthoma elasticum (PXE) has been associated with low levels of carboxylated matrix gla protein (MGP), most likely as a consequence of reduced vitamin K (vit K) availability. Unexpectedly, vit K supplementation does not exert beneficial effects on soft connective tissue mineralization in the PXE animal model. To understand the effects of vit K supplementation and in the attempt to interfere with pathways leading to the accumulation of calcium and phosphate within PXE-mineralized soft connective tissues, we have conducted in vitro studies on dermal fibroblasts isolated from control subjects and from PXE patients. Cells were cultured in standard conditions and in calcifying medium (CM) in the presence of vit K1 and K2, or levamisole, an alkaline phosphatase (ALP) inhibitor. Control and PXE fibroblasts were characterized by a similar dose-dependent uptake of both vit K1 and vit K2, thus promoting a significant increase of total protein carboxylation in all cell lines. Nevertheless, MGP carboxylation remained much less in PXE fibroblasts. Interestingly, PXE fibroblasts exhibited a significantly higher ALP activity. Consistently, the mineralization process induced in vitro by a long-term culture in CM appeared unaffected by vit K, whereas it was abolished by levamisole.

  14. Snow chemistry of high altitude glaciers in the French Alps

    NASA Astrophysics Data System (ADS)

    Maupetit, François; Delmas, Robert J.

    1994-09-01

    Snow samples were collected as snowcores in the accumulation zone of four high altitude glaciers (2980 3540m.a.s.l.) from each of the 4 highest mountain areas of the French Alps, during 3 consecutive years: 1989, 1990 and 1991. Sampling was performed in spring (˜ May), before the onset of late spring summer percolation. The accumulated snow therefore reflects winter and spring conditions. A complementary sampling of fresh-snow was performed on an event basis, on one of the studied glaciers, in 1990 and 1991. All samples were analysed for major ions (but also for total formate and acetate in fresh-snow samples) using ion chromatography. The acidity-alkalinity was accurately determined with a titration technique. The ion balance of alpine snow has been achieved from those analyses. High alpine snow is slightly acid (H+~ 3 20 μeq 1-1), but is episodically affected by alkaline saharan dust events. The different sources (pollution, seasalt and soil dust) affecting the impurity content of snow were identified using principal component analysis. The measured free acidity, mainly from anthropogenic origin, originates from nitric acid scavenging while sulfuric acidity is partially neutralized by atmospheric ammonia and by alkaline soil dust derived species, the contribution of hydrochloric acid being negligible. All ions exhibit higher concentrations in spring than in winter snow, indicating most likely the influence of increased vertical transport from the lower troposphere at this time. The transport of saharan dust is described through three major events reaching the Alps during March 1990 and 1991. Very high concentrations of Ca2+ and HCO3- were measured in corresponding samples, indicating that

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

    SciTech Connect

    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%, 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).

  16. 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.

  17. 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. Alps

    Atmospheric Science Data Center

    2013-04-17

    ... that run the length of Italy in the center, and the islands of Corsica and the larger Sardinia off Italy's west coast. The city of ... of the boot at the northwest end of the Adriatic Sea. The barrier islands across the Gulf of Venice are barely visible. Trieste is on the ...

  3. Effect of apatite formation of biphasic calcium phosphate ceramic (BCP) on osteoblastogenesis using simulated body fluid (SBF) with or without bovine serum albumin (BSA).

    PubMed

    Huang, Li; Zhou, Bo; Wu, Huayu; Zheng, Li; Zhao, Jinmin

    2017-01-01

    Although biphasic calcium phosphate ceramic (BCP) holds promise in therapy of bone defect, surface mineralization prior to implantation may improve the bioactivity to better integrate with the host. Immersion in simulated body fluid (SBF) and bovine serum albumin-simulated body fluid (BSA-SBF) are common methods to form apatite interface layer. This study was intended to investigate the effect of SBF and BSA-SBF treatment on the bioactivity of BCP in vitro. In this study, osteoblasts were grown on BCP with or without treatment of SBF or BSA-SBF, and detected with general observation, scanning electron microscope (SEM), cell proliferation assay, morphology observation, viability assay, alkaline phosphatase (ALP) activity assay, and osteogenic specific gene expression of alkaline phosphatase (ALPL), bone gamma-carboxyglutamate (gla) protein (BGLAP), bone morphogenetic protein 2 (BMP2), bone sialoprotein (BSP), type I collagen (COLI) and runt-related transcription factor 2 (RUNX2) after culture of 2, 5 and 8days. As the results shown, BCP pre-incubated in SBF and BSA-SBF up-regulated ALP activity and osteogenic related genes and proteins, which testified the positive effect of SBF and BSA-SBF. Especially, BSA-SBF enhanced the cell growth significantly. This study indicated that treatment by BSA-SBF is of importance for BCP before clinical application.

  4. 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.

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

    PubMed

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

    2015-12-01

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

  6. Cells responding to surface structure of calcium phosphate ceramics for bone regeneration.

    PubMed

    Zhang, Jingwei; Sun, Lanying; Luo, Xiaoman; Barbieri, Davide; de Bruijn, Joost D; van Blitterswijk, Clemens A; Moroni, Lorenzo; Yuan, Huipin

    2017-02-08

    Surface structure largely affects the inductive bone-forming potential of calcium phosphate (CaP) ceramics in ectopic sites and bone regeneration in critical-sized bone defects. Surface-dependent osteogenic differentiation of bone marrow stromal cells (BMSCs) partially explained the improved bone-forming ability of submicron surface structured CaP ceramics. In this study, we investigated the possible influence of surface structure on different bone-related cells, which may potentially participate in the process of improved bone formation in CaP ceramics. Besides BMSCs, the response of human brain vascular pericytes (HBVP), C2C12 (osteogenic inducible cells), MC3T3-E1 (osteogenic precursors), SV-HFO (pre-osteoblasts), MG63 (osteoblasts) and SAOS-2 (mature osteoblasts) to the surface structure was evaluated in terms of cell proliferation, osteogenic differentiation and gene expression. The cells were cultured on tricalcium phosphate (TCP) ceramics with either micron-scaled surface structure (TCP-B) or submicron-scaled surface structure (TCP-S) for up to 14 days, followed by DNA, alkaline phosphatase (ALP) and quantitative polymerase chain reaction gene assays. HBVP were not sensitive to surface structure with respect to cell proliferation and osteogenic differentiation, but had downregulated angiogenesis-related gene expression (i.e. vascular endothelial growth factor) on TCP-S. Without additional osteogenic inducing factors, submicron-scaled surface structure enhanced ALP activity and osteocalcin gene expression of human (h)BMSCs and C2C12 cells, favoured the proliferation of MC3T3-E1, MG63 and SAOS-2, and increased ALP activity of MC3T3-E1 and SV-HFO. The results herein indicate that cells with osteogenic potency (either osteogenic inducible cells or osteogenic cells) could be sensitive to surface structure and responded to osteoinductive submicron-structured CaP ceramics in cell proliferation, ALP production or osteogenic gene expression, which favour bone

  7. 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

  8. Updated Understanding of Autoimmune Lymphoproliferative Syndrome (ALPS).

    PubMed

    Li, Pu; Huang, Ping; Yang, Ye; Hao, Mu; Peng, Hongwei; Li, Fei

    2016-02-01

    Autoimmune lymphoproliferative syndrome (ALPS), a disorder characterized by immune dysregulation due to disrupted lymphocyte homeostasis, is mainly resulted from the mutations in FAS-mediated apoptotic pathway. In addition, other mutations of the genes such as Fas-ligand (FASLG), Caspase 10 (CASP10) and Caspase 8 (CASP8), NRAS and KRAS have also been observed in a small number of patients with ALPS or ALPS-related disorders. However, approximately 20-30% of patients with ALPS have unidentified defect. Its clinical manifestations observed in multiple family members include unexplained lymphadenopathy, hepatosplenomegaly, autoimmune cytopenias such as thrombocytopenia, neutropenia, and anemia due to excessive production of antibodies by lymphocytes, elevated number of double-negative T (DNT) cells, and increased risk of lymphoma. As a very rare disease, ALPS was first characterized in the early 1990s. More than 300 families with hereditary ALPS have been reported till now; nearly 500 patients from these families have been studied and followed worldwide over the last 20 years. ALPS has historically considered as a primary immune defect presenting in early childhood, however, recent studies have shown that it may be more common than previous thought because adult onset presentation is increasingly becoming recognized and more adult ALPS patients are diagnosed. The new genetic and biological insights have improved the understanding of ALPS and a number of targeted therapeutic strategies such as mycophenolate mofetil, sirolimus, and pentostatin have been successfully applied in ALPS patients with promising treatment efficacy. This article comprehensively reviews the clinical and laboratory manifestations, new research advances in the molecular pathogenesis, diagnosis and treatments of this disorder.

  9. Osteogenic and angiogenic potentials of monocultured and co-cultured human-bone-marrow-derived mesenchymal stem cells and human-umbilical-vein endothelial cells on three-dimensional porous beta-tricalcium phosphate scaffold.

    PubMed

    Kang, Yunqing; Kim, Sungwoo; Fahrenholtz, Monica; Khademhosseini, Ali; Yang, Yunzhi

    2013-01-01

    The use of biodegradable beta-tricalcium phosphate (β-TCP) scaffolds holds great promise for bone tissue engineering. However, the effects of β-TCP on bone and endothelial cells are not fully understood. This study aimed to investigate cell proliferation and differentiation of mono- or co-cultured human-bone-marrow-derived mesenchymal stem cells (hBMSCs) and human-umbilical-vein endothelial cells (HUVECs) on a three-dimensional porous, biodegradable β-TCP scaffold. In co-culture studies, the ratios of hBMSCs:HUVECs were 5:1, 1:1 and 1:5. Cellular morphologies of HUVECs, hBMSCs and co-cultured HUVECs/hBMSCs on the β-TCP scaffolds were monitored using confocal and scanning electron microscopy. Cell proliferation was monitored by measuring the amount of double-stranded DNA (dsDNA) whereas hBMSC and HUVEC differentiation was assessed using the osteogenic and angiogenic markers, alkaline phosphatase (ALP) and PECAM-1 (CD31), respectively. Results show that HUVECs, hBMSCs and hBMSCs/HUVECs adhered to and proliferated well on the β-TCP scaffolds. In monoculture, hBMSCs grew faster than HUVECs on the β-TCP scaffolds after 7 days, but HUVECs reached similar levels of proliferation after 14 days. In monoculture, β-TCP scaffolds promoted ALP activities of both hBMSCs and HUVECs when compared to those grown on tissue culture well plates. ALP activity of cells in co-culture was higher than that of hBMSCs in monoculture. Real-time polymerase chain reaction results indicate that runx2 and alp gene expression in monocultured hBMSCs remained unchanged at days 7 and 14, but alp gene expression was significantly increased in hBMSC co-cultures when the contribution of individual cell types was not distinguished.

  10. Tertiary LCT and NYF pegmatites of the central Alps

    NASA Astrophysics Data System (ADS)

    Guastoni, Alessandro; Pennacchioni, Giorgio

    2014-05-01

    A large field (about 100 km in E-W length and 15 km in N-S thickness) of Oligocene pegmatites extends in the central Alps from the Bergell pluton (to the east) to the Ossola Valley (to the west) within the Alpine nappes north of the Periadriatic Lineament. The pegmatite field geographically overlaps (i) the highest temperature domain of the Lepontine Barrovian metamorphic dome, and (ii) the zone of Alpine migmatization. Most pegmatites have a simple mineralogy consisting of K-feldspar, quartz and muscovite, but a minor amount (< 5%) includes Sn-Nb-Ta-Y-REE-U oxides, Y-REE phosphates, Mn-Fe-phosphates, Ti-Zr-silicates, Be-Y-REE-silicates, garnet (almandine-spessartine), and schorl-dravite-fluorelbaite tourmaline. Major and trace elements geochemistry of pegmatite bulk rock, rock-forming and accessory minerals allows the distinction of different pegmatite populations ranging from NYF (niobium, yttrium, fluorine) to LCT (lithium, cesium, tantalum) pegmatites, or mixed LCT-NYF ones. Actually, LCT pegmatites of the Central Alps did not reach a high degree of geochemical evolution. In the Codera Valley (on the western side of the Bergell pluton) LCT and NYF pegmatites are respectively hosted in tonalites and granodiorites; these pegmatites include the most evolved types which contain Mn-fluorelbaite, Mn-phosphates, pink-beryl and Cs-Rb-rich feldspar. From the structural point of view 2 main types of pegmatites can be distinguished: (i) pegmatites that were involved in ductile deformation, and (ii) pegmatite crosscutting the ductile structures of the SSB. Many pegmatites from Codera Valley belong to the first structural type: they were at emplaced at relatively high ambient temperatures (> 450 °C) and locally show pervasive recrystallization of quartz. More to the east (Mesolcina and Bodengo Valleys) the main set of pegmatites crosscut the ductile deformation structures of the SSB, but the area also includes an earlier generation of boudinaged and folded dykes. The

  11. 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

  12. 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.

  13. Histochemical localization of alkaline phosphatase activity in decalcified bone and cartilage.

    PubMed

    Miao, Dengshun; Scutt, Andrew

    2002-03-01

    We have developed methodology that enables alkaline phosphatase (ALP) to be histochemically stained reproducibly in decalcified paraffin-embedded bone and cartilage of rodents. Proximal tibiae and fourth lumbar vertebrae were fixed in periodate-lysine-paraformaldehyde (PLP) fixative, decalcified in an EDTA-G solution, and embedded in paraffin. In the articular cartilage of the proximal tibia, ALP activity was localized to the hypertrophic chondrocytes and cartilage matrix of the deep zone and the maturing chondrocytes of the intermediate zone. The cells and matrix in the superficial zone did not exhibit any enzyme activity. In tibial and vertebral growth plates, a progressive increase in ALP expression was seen in chondrocytes and cartilage matrix, with activity being weakest in the proliferative zone, higher in the maturing zone, and highest in the hypertrophic zone. In bone tissue, ALP activity was detected widely in pre-osteoblasts, osteoblasts, lining cells on the surface of trabeculae, some newly embedded osteocytes, endosteal cells, and subperiosteal cells. In areas of new bone formation, ALP activity was detected in osteoid. In the bone marrow, about 20% of bone marrow cells expressed ALP activity. In adult rats, the thickness of the growth plates was less and ALP activity was enhanced in maturing and hypertrophic chondrocytes, cartilage matrix in the hypertrophic zone, and primary spongiosa. This is the first time that ALP activity has been successfully visualized histochemically in decalcified, paraffin-embedded mineralized tissues. This technique should prove to be a very convenient adjunct for studying the behavior of osteoblasts during osteogenesis.

  14. Lectin affinity electrophoresis of serum alkaline phosphatase in metastasized breast cancer.

    PubMed

    Le Bricon, Thierry; Gay-Bellile, Cécile; Cottu, Paul; Benlakehal, Mourad; Guillon, Hélène; Houzé, Pascal

    2010-01-01

    The use of serum alkaline phosphatase (ALP) isoenzymes as markers of breast cancer metastases and treatment efficacy has received little attention. Twenty-six breast cancer women (56+/-13 years, all post-menopausal) were prospectively evaluated during their first and third course of chemotherapy (4-week interval). Serum samples were analyzed for ALP isoenzymes (bone, liver, and intestine) using a lectin affinity electrophoresis kit (Hydragel 15 ISO-PAL, Sebia) adapted on a semi-automated Hydrasys system (Sebia). Results were compared with imaging techniques for the presence of metastases; bone ALP isoenzyme (B-ALP) results were compared with C-Terminal degradation products of type I collagen (S-CTX) (CrossLaps, IDS Nordic). Serum B-ALP, but not S-CTX, confirmed the presence of bone metastases (BM) (n=15) with 67/100% sensitivity/specificity (using a 69 UI/L ROC cut-off); ROC AUC was 0.806 (P=0.0004) (NS for S-CTX). Chemotherapy reduced serum B-ALP by 24% over 4 weeks (P=0.0012); there was no change for S-CTX. There was no specific clinical pattern for other ALP isoenzymes (liver and intestine). In conclusion, serum B-ALP, but not S-CTX, could help confirm the presence of BM in breast cancer patients.

  15. Functional interrelationships in the alkaline phosphatase superfamily: phosphodiesterase activity of Escherichia coli alkaline phosphatase.

    PubMed

    O'Brien, P J; Herschlag, D

    2001-05-15

    Escherichia coli alkaline phosphatase (AP) is a proficient phosphomonoesterase with two Zn(2+) ions in its active site. Sequence homology suggests a distant evolutionary relationship between AP and alkaline phosphodiesterase/nucleotide pyrophosphatase, with conservation of the catalytic metal ions. Furthermore, many other phosphodiesterases, although not evolutionarily related, have a similar active site configuration of divalent metal ions in their active sites. These observations led us to test whether AP could also catalyze the hydrolysis of phosphate diesters. The results described herein demonstrate that AP does have phosphodiesterase activity: the phosphatase and phosphodiesterase activities copurify over several steps; inorganic phosphate, a strong competitive inhibitor of AP, inhibits the phosphodiesterase and phosphatase activities with the same inhibition constant; a point mutation that weakens phosphate binding to AP correspondingly weakens phosphate inhibition of the phosphodiesterase activity; and mutation of active site residues substantially reduces both the mono- and diesterase activities. AP accelerates the rate of phosphate diester hydrolysis by 10(11)-fold relative to the rate of the uncatalyzed reaction [(k(cat)/K(m))/k(w)]. Although this rate enhancement is substantial, it is at least 10(6)-fold less than the rate enhancement for AP-catalyzed phosphate monoester hydrolysis. Mutational analysis suggests that common active site features contribute to hydrolysis of both phosphate monoesters and phosphate diesters. However, mutation of the active site arginine to serine, R166S, decreases the monoesterase activity but not the diesterase activity, suggesting that the interaction of this arginine with the nonbridging oxygen(s) of the phosphate monoester substrate provides a substantial amount of the preferential hydrolysis of phosphate monoesters. The observation of phosphodiesterase activity extends the previous observation that AP has a low level of

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

    SciTech Connect

    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.

  17. The Effect of Alendronate Loaded Biphasic Calcium Phosphate Scaffolds on Bone Regeneration in a Rat Tibial Defect Model

    PubMed Central

    Park, Kwang-Won; Yun, Young-Pil; Kim, Sung Eun; Song, Hae-Ryong

    2015-01-01

    This study investigated the effect of alendronate (Aln) released from biphasic calcium phosphate (BCP) scaffolds. We evaluated the in vitro osteogenic differentiation of Aln/BCP scaffolds using MG-63 cells and the in vivo bone regenerative capability of Aln/BCP scaffolds using a rat tibial defect model with radiography, micro-computed tomography (CT), and histological examination. In vitro studies included the surface morphology of BCP and Aln-loaded BCP scaffolds visualized using field-emission scanning electron microscope, release kinetics of Aln from BCP scaffolds, alkaline phosphatase (ALP) activity, calcium deposition, and gene expression. The in vitro studies showed that sustained release of Aln from the BCP scaffolds consisted of porous microstructures, and revealed that MG-63 cells cultured on Aln-loaded BCP scaffolds showed significantly increased ALP activity, calcium deposition, and gene expression compared to cells cultured on BCP scaffolds. The in vivo studies using radiograph and histology examination revealed abundant callus formation and bone maturation at the site in the Aln/BCP groups compared to the control group. However, solid bony bridge formation was not observed at plain radiographs until 8 weeks. Micro-CT analysis revealed that bone mineral density and bone formation volume were increased over time in an Aln concentration-dependent manner. These results suggested that Aln/BCP scaffolds have the potential for controlling the release of Aln and enhance bone formation and mineralization. PMID:26561810

  18. The Effect of Covalently Immobilized FGF-2 on Biphasic Calcium Phosphate Bone Substitute on Enhanced Biological Compatibility and Activity.

    PubMed

    Moon, Kyung-Suk; Choi, Eun-Joo; Oh, Seunghan; Kim, Sungtae

    2015-01-01

    The purpose of this research was to covalently graft fibroblast growth factor 2 (FGF-2) onto biphasic calcium phosphate (BCP) via a bifunctional cross-linker technique and to estimate the optimal dose of FGF-2 resulting in the best osteogenic differentiation of human mesenchymal stem cells (hMSCs). SEM observation revealed that the surface of the 100 ng FGF-2 coated BCP was completely covered with the nanoparticles expected to be from the silane coupling agent. XRD, FT-IR, and XPS analysis showed that silane treatment, bifunctional cross-linker coating, and FGF-2 covalent grafts were conducted successfully without deforming the crystalline structure of BCP. An MTT assay demonstrated that FGF-2 coated BCP had good biocompatibility, regardless of the concentration of FGF-2, after 24 or 48 h of incubation. An alkaline phosphatase (ALP) activity assay (14 days of incubation) and the ALP gene expression level of real-time PCR analysis (7 days of incubation) revealed that 50, 100, and 200 ng FGF-2 coated BCP induced the highest activities among all experimental groups and control group (P < 0.05). Thus, low concentrations of FGF-2 facilitated excellent osteogenesis and were effective at enhancing osteogenic potential. Also, the bifunctional cross-linker technique is expected to be a more feasible way to induce osteogenic differentiation while minimizing the risk of FGF-2 overdose.

  19. Bioactivity and cytocompatibility of dicalcium phosphate/poly (amino acid) biocomposite with degradability

    NASA Astrophysics Data System (ADS)

    Zhang, Yunfei; Shan, Wenpeng; Li, Xiangde; Wei, Jie; Li, Hong; Ma, Jian; Yan, Yonggang

    2012-01-01

    A bioactive composite of dicalcium phosphate (DCP) and poly (amino acid) (PAA) was fabricated, and the in vitro bioactivity, degradability, and cellular responses to the DCP/PAA composite (DPC) were investigated as compared to PAA. Apatite formation on DPC surfaces occurred after immersion into simulated body fluid (SBF) for 7 days, but not on the surface of PAA. The weight loss ratio of DPC could reach 18.6 ± 0.3 wt% after soaking into phosphate buffered saline (PBS) for 2 months, which was higher than PAA (11.0 ± 0.2 wt%). Cell attachment and proliferation of MG-63 cells on DPC was obviously higher than on PAA. Moreover, the cells spread and formed confluent layer on the DPC surfaces. The alkaline phosphatase activity (ALP) of the cells on DPC was significantly greater than PAA at day 5 and day 7. The results suggested that introducing DCP into PAA makes the composite bioactive and more degradable, and meanwhile enhances osteoblast-like cells attach, proliferation and osteogenic differentiation.

  20. Monte Carlo Simulations of Phosphate Polyhedron Connectivity in Glasses

    SciTech Connect

    ALAM,TODD M.

    1999-12-21

    Monte Carlo simulations of phosphate tetrahedron connectivity distributions in alkali and alkaline earth phosphate glasses are reported. By utilizing a discrete bond model, the distribution of next-nearest neighbor connectivities between phosphate polyhedron for random, alternating and clustering bonding scenarios was evaluated as a function of the relative bond energy difference. The simulated distributions are compared to experimentally observed connectivities reported for solid-state two-dimensional exchange and double-quantum NMR experiments of phosphate glasses. These Monte Carlo simulations demonstrate that the polyhedron connectivity is best described by a random distribution in lithium phosphate and calcium phosphate glasses.

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

    SciTech Connect

    Khokher, M.A.; Dandona, P. )

    1990-11-01

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

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

    PubMed Central

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

    2009-01-01

    A one-step, single use, disposable Alkaline Phosphatase (ALP) biosensor has been developed. It is based on the detection of phenol produced by an ALP enzymatic reaction. It can operate at 25 °C in a pH 10 medium. It measures ALP of 0–300 IU/L. The permissible concentrations of glucose, ascorbic acid and urea without interference are 10 mM/L, 5 mg/L and 400 mg/L, respectively. Experimental results are compared to those obtained by spectrophotometric measurements in bovine serum. Excellent linearity between the biosensor outputs and the ALP concentrations exists. The agreement between the measurements of this biosensor and the spectrophotometer is also outstanding. PMID:22291532

  3. The Accelerated Learning Program (ALP) 2000-01: Student Participation and Effectiveness. ALP Report.

    ERIC Educational Resources Information Center

    Baenen, Nancy; Yaman, Kimberly; Lindblad, Mark

    The Accelerated Learning Program (ALP) is the major initiative that the Wake County Public School System (WCPSS), North Carolina, is using to help all students reach grade level performance in reading and mathematics. This report focuses on student participation rates and the impact of the ALP program. Data are from a variety of sources. In the…

  4. Determination of urinary and serum beta-glucuronidase and alkaline phosphatase in various renal disease and kidney rejection transplanted patients.

    PubMed

    Refaie, M O; Abo-Zaid, H; Gomma, N A; Aboul-Enein, H Y

    2000-05-01

    Beta-glucuronidase (beta-Glu) and alkaline phosphatase (ALP) were evaluated in serum and urine in 50 subjects classified into five equal groups. Group I was control healthy subjects, while groups II, III, IV, and V were patients with nephritic syndrome, pyelonephritis, kidney rejection, and end-stage renal disease, respectively. Urinary beta-Glu was significantly elevated in all four groups; while serum enzyme showed no change. On the other hand, serum ALP showed a significant elevation in all abnormal groups. Accordingly, urinary beta-Glu and serum ALP could be used as diagnostic markers for various renal diseases.

  5. [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.

  6. 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

  7. Osteoblastic alkaline phosphatase mRNA is stabilized by binding to vimentin intermediary filaments.

    PubMed

    Schmidt, Yvonne; Biniossek, Martin; Stark, G Björn; Finkenzeller, Günter; Simunovic, Filip

    2015-03-01

    Vascularization is essential in bone tissue engineering and recent research has focused on interactions between osteoblasts (hOBs) and endothelial cells (ECs). It was shown that cocultivation increases the stability of osteoblastic alkaline phosphatase (ALP) mRNA. We investigated the mechanisms behind this observation, focusing on mRNA binding proteins. Using a luciferase reporter assay, we found that the 3'-untranslated region (UTR) of ALP mRNA is necessary for human umbilical vein endothelial cells (HUVEC)-mediated stabilization of osteoblastic ALP mRNA. Using pulldown experiments and nanoflow-HPLC mass spectrometry, vimentin was identified to bind to the 3'-UTR of ALP mRNA. Validation was performed by Western blotting. Functional experiments inhibiting intermediate filaments with iminodipropionitrile and specific inhibition of vimentin by siRNA transfection showed reduced levels of ALP mRNA and protein. Therefore, ALP mRNA binds to and is stabilized by vimentin. This data add to the understanding of intracellular trafficking of ALP mRNA, its function, and have possible implications in tissue engineering applications.

  8. The ALP-Enigma protein ALP-1 functions in actin filament organization to promote muscle structural integrity in Caenorhabditis elegans.

    PubMed

    Han, Hsiao-Fen; Beckerle, Mary C

    2009-05-01

    Mutations that affect the Z-disk-associated ALP-Enigma proteins have been linked to human muscular and cardiac diseases. Despite their clear physiological significance for human health, the mechanism of action of ALP-Enigma proteins is largely unknown. In Caenorhabditis elegans, the ALP-Enigma protein family is encoded by a single gene, alp-1; thus C. elegans provides an excellent model to study ALP-Enigma function. Here we present a molecular and genetic analysis of ALP-Enigma function in C. elegans. We show that ALP-1 and alpha-actinin colocalize at dense bodies where actin filaments are anchored and that the proper localization of ALP-1 at dense bodies is dependent on alpha-actinin. Our analysis of alp-1 mutants demonstrates that ALP-1 functions to maintain actin filament organization and participates in muscle stabilization during contraction. Reducing alpha-actinin activity enhances the actin filament phenotype of the alp-1 mutants, suggesting that ALP-1 and alpha-actinin function in the same cellular process. Like alpha-actinin, alp-1 also interacts genetically with a connectin/titin family member, ketn-1, to provide mechanical stability for supporting body wall muscle contraction. Taken together, our data demonstrate that ALP-1 and alpha-actinin function together to stabilize actin filaments and promote muscle structural integrity.

  9. Phosphate salts

    MedlinePlus

    ... sodium if you have heart disease. Fluid retention (edema): Avoid using phosphate salts that contain sodium if ... heart failure, or other conditions that can cause edema. High levels of calcium in the blood (hypercalcemia): ...

  10. Hierarchically microporous/macroporous scaffold of magnesium-calcium phosphate for bone tissue regeneration.

    PubMed

    Wei, Jie; Jia, Junfeng; Wu, Fan; Wei, Shicheng; Zhou, Huanjun; Zhang, Hongbo; Shin, Jung-Woog; Liu, Changsheng

    2010-02-01

    Hierarchically 3D microporous/macroporous magnesium-calcium phosphate (micro/ma-MCP) scaffolds containing magnesium ammonium phosphate hexahydrate [NH(4)MgPO(4).6H(2)O] and hydroxyapatite [Ca(10)(PO(4))(6)(OH)(2)] were fabricated from cement utilizing leaching method in the presence of sodium chloride (NaCl) particles and NaCl saturated water solution. NaCl particles produced macroporosity, and NaCl solution acted as both cement liquid and porogens, inducing the formation of microporosity. The micro/ma-MCP scaffolds with porosities varied from 52 to 78% showed well interconnected and open macropores with the sizes of 400-500 microm, and degradation of the scaffolds was significantly enhanced in Tris-HCl solution compared with macroporous MCP (ma-MCP) and corresponding calcium phosphate cement (CPC) scaffolds. Cell attachment and proliferation of MG(63) on micro/ma-MCP were significantly better than ma-MCP and CPC scaffolds because of the presence of microporosity, which enhanced the surface area of the scaffolds. Moreover, the alkaline phosphatase (ALP) activity of the MG(63) cells on micro/ma-MCP was significantly higher than ma-MCP and CPC scaffolds at 7 days, and the MG(63) cells with normal phenotype spread well and formed confluent layers across the macroporous walls of the micro/ma-MCP scaffolds. Histological evaluation confirmed that the micro/ma-MCP scaffolds improved the efficiency of new bone regeneration, and exhibited excellent biocompatibility, biodegradability and faster and more effective osteogenesis in vivo.

  11. Updated core libraries of the ALPS project

    NASA Astrophysics Data System (ADS)

    Gaenko, A.; Antipov, A. E.; Carcassi, G.; Chen, T.; Chen, X.; Dong, Q.; Gamper, L.; Gukelberger, J.; Igarashi, R.; Iskakov, S.; Könz, M.; LeBlanc, J. P. F.; Levy, R.; Ma, P. N.; Paki, J. E.; Shinaoka, H.; Todo, S.; Troyer, M.; Gull, E.

    2017-04-01

    The open source ALPS (Algorithms and Libraries for Physics Simulations) project provides a collection of physics libraries and applications, with a focus on simulations of lattice models and strongly correlated systems. The libraries provide a convenient set of well-documented and reusable components for developing condensed matter physics simulation code, and the applications strive to make commonly used and proven computational algorithms available to a non-expert community. In this paper we present an updated and refactored version of the core ALPS libraries geared at the computational physics software development community, rewritten with focus on documentation, ease of installation, and software maintainability.

  12. Anodes for alkaline electrolysis

    DOEpatents

    Soloveichik, Grigorii Lev

    2011-02-01

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

  13. Prognostic value of combined preoperative lactate dehydrogenase and alkaline phosphatase levels in patients with resectable pancreatic ductal adenocarcinoma

    PubMed Central

    Ji, Fei; Fu, Shun-Jun; Guo, Zhi-Yong; Pang, Hui; Ju, Wei-Qiang; Wang, Dong-Ping; Hua, Yun-Peng; He, Xiao-Shun

    2016-01-01

    Abstract Serum enzymes, including lactate dehydrogenase (LDH) and alkaline phosphatase (ALP), have recently been reported to play important roles in tumor growth. Increases in LDH and ALP have been confirmed to predict poor prognosis in patients with various cancers. However, their prognostic value in pancreatic cancer has not been well studied. Therefore, we reviewed the preoperative data on LDH and ALP in 185 pancreatic ductal adenocarcinoma (PDAC) patients who underwent surgery between July 2005 and December 2010 to explore the prognostic value of these markers. The cutoff points were determined based on the upper limit of their normal values. The Chi-square test was used to analyze the relationships between LDH/ALP and clinical characteristics. Univariate and multivariate analyses were performed to identify the predictive value of the above factors for disease-free survival (DFS) and overall survival (OS). We found that elevation of LDH was related to carbohydrate antigen 19-9 (CA19-9), lymph node involvement, tumor size, TNM, distant metastasis, and recurrence. Additionally, ALP was correlated to perineural invasion. After multivariate analysis, LDH and ALP were identified as independent prognostic factors for DFS and OS, and elevation of LDH/ALP was correlated with poor DFS and OS. Notably, there was a positive correlation between LDH and ALP. The predictive power of LDH combined with ALP was more sensitive than that of either one alone. Therefore, we conclude that the preoperative LDH and ALP values are prognostic factors for PADC, and the prognostic accuracy of testing can be enhanced by the combination of LDH and ALP PMID:27399091

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

    PubMed

    Homaei, Ahmad

    2017-02-01

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

  15. Alkaline phosphatase of Physarum polycephalum is insoluble.

    PubMed

    Furuhashi, Kiyoshi

    2008-02-01

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

  16. Purification and characterization of Ulva pertusa Kjellm alkaline phosphatase.

    PubMed

    Yang, Dong; Wang, Jingyun; Bao, Yongming; An, Lijia

    2003-05-01

    The activity of alkaline phosphatase (ALP, EC 3.1.3.1.) was found in seaweeds, including five kinds of green alga, eighteen kinds of red alga, and six kinds of brown alga, collected from the seaside of Dalian in China. The enzyme was purified 1230-fold from Ulva pertusa Kjellm. It had a specific activity of 48.6 U/mg protein and was proven to be homogeneous by SDS-PAGE with a subunit molecular mass of 19.5 kDa. The activity of ALP peaked at pH9.8, and was completely inhibited by DTT and partly by NBS. The Michaelis-Menten constant Km and the maximum reaction velocity Vmax, at pH 9.8 and 37 degrees C were 0.950 mM and 5.00 microM/min, respectively.

  17. GOMEX08 - High Frequency Acoustic Propagation over the Alabama Alps

    DTIC Science & Technology

    2010-06-01

    of a group of reef -like structures collectively known as the Pinnacles . In particular, the Alabama Alps rises to about 15-17m above a relatively...the general area of the Pinnacles and the specific Ala- bama Alps experimental site of this paper. The Alabama Alps region (Fig. 1, lower left) was...chosen for its simplicity as compared with the Scamp Reef region (Fig. 1, upper right). The experimental geometry at the Alps is overlaid on the

  18. Dyscalculia and the Minicalculator: The ALP Program.

    ERIC Educational Resources Information Center

    Block, Gerald H.

    1980-01-01

    The author points out the usefulness of the ALP minicalculator program with learning disabled learners and introduces a novel aspect of the program--a new method of dealing with fractions via the calculator in which decimal answers are converted to equivalent fractions. (SBH)

  19. An economics figure of merit in ALPS

    SciTech Connect

    Shatilla, Y.A.

    2000-07-01

    One of the most pressing issues facing the deregulated nuclear electric power industry is its economic competitiveness when compared to other sources of electrical power. Traditionally, finding the optimum loading pattern (LP) that meets all the safety and operational objective functions and at the same time produces the most attractive economical solutions is an iterative process. This is because (a) LP search tools usually lack the capability to generate equilibrium solutions and (b) economics objective functions are hard to include in the search process. In this paper, the Westinghouse Advanced Loading Pattern Search code (ALPS) has been demonstrated to successfully find LPs that meet user-defined operational and safety as well as economics objectives. This has been made possible by the development of TULIP language that allows the integration of external procedures into the search process of the main program, ALPS. In the example given, an economic figure of merit (EFM) has been defined and included via TULIP script into the fuel management optimization problem of a three-loop Westinghouse core operating an 18-month cycle. The LPs found by ALPS exhibit a clear trend of meeting and, in some cases, exceeding the EFM objective function defined for the ALPS search process a priori.

  20. 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)

  1. Inhibition kinetics of acid and alkaline phosphatases by atrazine and methomyl pesticides.

    PubMed

    El-Aswad, Ahmed F; Badawy, Mohamed E I

    2015-01-01

    The main objective of this work was to investigate the kinetic characteristics of acid and alkaline phosphatases isolated from different sources and to study the effects of the herbicide atrazine and insecticide methomyl on the activity and kinetic properties of the enzymes. Acid phosphatase (ACP) was isolated from the tomato plant (Solanum lycopersicum L. var. lycopersicum); alkaline phosphatase (ALP) was isolated from two sources, including mature earthworms (Aporrectodea caliginosa) and larvae of the Egyptian cotton leafworm (Spodoptera littoralis). The specific activities of the enzymes were 33.31, 5.56 and 0.72 mmol substrate hydrolyzed per minute per milligram protein for plant ACP, earthworms ALP and cotton leafworm ALP, respectively. The inhibition kinetics indicated that atrazine and methomyl caused competitive-non-competitive inhibition of the enzymes. The relationships between estimates of K(m) and V(max) calculated from the Michaelis-Menten equation have been explored. The extent of the inhibition was different, as estimated by the values of the inhibition constant Ki that were found to be 3.34 × 10(-3), 1.12 × 10(-2) and 1.07 × 10(-2) mM for plant ACP, earthworms ALP and cotton leafworm ALP, respectively, with methomyl. In the case of atrazine, K(i) were found to be 8.99 × 10(-3), 3.55 × 10(-2) and 1.36 × 10(-2) mM for plant ACP, earthworms ALP and cotton leafworm ALP, respectively.

  2. 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.

  3. Physiological aspects of alkaline phosphatase in selected cyanobacteria.

    PubMed

    Doonan, B B; Jensen, T E

    1980-01-01

    The alkaline phosphatase of Plectonema boryanum shows a considerable increase in activity following placement of the cells in a phosphate free medium. Five days of phosphate starvation result in a 14-fold increase of alkaline phosphatase activity. Growth in the presence of inhibitors of transcription and translation indicate that the synthesis of the enzyme is de novo. Orthophosphate causes an immediate inhibition of enzyme activity. Enzyme was extracted from P. boryanum with lysozyme or polymyxin B treatment in order to make comparative studies of cell bound and cell free enzyme. Of several enzyme specific inhibitors tested, mercuric chloride was the most effective. Temperature studies showed that the cell bound enzyme was most active at 40 degrees C while the cell free enzyme was most active at 70 degrees C. The pH optimum was 9 for the cell free enzyme, and 8.8 for the cell bound. The enzyme was tested to determine if it could hydrolyse a number of different organic compounds. It hydrolysed p-nitrophenol phosphate 100%, fructose-6-phosphate 45%, beta-glycerol phosphate 25% and other compounds to a lesser degree. Of seventeen other Cyanobacteria tested for alkaline phosphatase, all were positive, and of these eleven were inducible for the enzyme. Ten of the isolates released some of the enzyme into the culture medium. Michaelis constants for the enzyme were also determined.

  4. Alkaline phosphatase-labeled macromolecular probe for sensitive chemiluminescence detection of proteins on a solid-phase membrane.

    PubMed

    Azam, Md Golam; Shibata, Takayuki; Kabashima, Tsutomu; Kai, Masaaki

    2011-09-01

    In the present study, we synthesized dextran (MW = ca. 2,000 kDa)-based macromolecular probes containing multiple molecules of alkaline phosphatase (ALP) as a signal-trigger enzyme and of biotin as an assembly mediator. The ALP and biotin molecules were covalently attached into the dextran backbone after the formation of aldehyde groups into the macromolecule by periodate oxidation. The synthesized probes contained 27-31 molecules of ALP in their macromolecules when 50-fold molar ratio of ALP to the dextran was used for the synthesis. These probes provided 14-20 times stronger chemiluminescence (CL) than that of the equimolar free ALP adsorbed on a nylon membrane. The velocity of the CL reaction of ALP-catalyzed adamantlyl-1,2-dioxetane substrate was improved from a slower emission (glow type) of CL to a faster one (flash type). The CL signal integrated for 2 min under strongly alkaline conditions (pH 13.0) was about ten times greater than that obtained by the conventional conditions (pH 9.5). Therefore, the synthesized macromolecular probe could be successfully utilized for the high-throughput CL detection of biotin-conjugated anti-rabbit IgG antibody with a lower detection limit of 880 amol per spot on the nylon membrane. This study provides analytical strategy for the rapid, convenient, and sensitive detection of target proteins in immunoassays.

  5. 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

  6. 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.

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

    PubMed

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

    2016-03-01

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

  8. 1-step versus 2-step immobilization of alkaline phosphatase and bone morphogenetic protein-2 onto implant surfaces using polydopamine.

    PubMed

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

    2013-08-01

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

  9. Serum Alkaline Phosphatase Level as a Prognostic Tool in Colorectal Cancer: A Study of 105 patients.

    PubMed

    Saif, M Wasif; Alexander, Dominik; Wicox, Charles M

    2005-01-01

    BACKGROUND: Serum alkaline phosphotase (ALP) levels are frequently elevated in patients with metastatic colorectal cancer (CRC). However, the significance of ALP in terms of detecting hepatic metastasis or prognosis is not well established. MATERIALS AND METHODS: Medical records of patients with CRC seen at University of Alabama at Birmingham (UAB) (1998-2002) were reviewed and statistical analysis was done to evaluate the significance of ALP as a prognostic tool. The normal range for ALP was quantified at 39 U/L to 117 U/L. Change in ALP levels over time (defined as time interval between two cycles; such as 4 weeks for Mayo regimen, 8 weeks for Roswell Park regimen and 6 weeks for IFL regimen) was categorized as large (120+ U/L), medium (20-119 U/L), and minimal (< 20 U/L). RESULTS: A total of 105 patients with eligible medical records were identified (Mean age: 59 yrs; 53% male; Staging: II: 43 patients, III: 31 patients, IV: 32 patients). Increasing ALP levels correlated with increasing stage (Mean: I = 116, II = 219, III = 302; P = 0.0003). ALP levels were elevated in 74% of patients with liver metastases (Mean, 290) and in 33% without liver metastases (Mean, 122) (P = 0.001). Patients with elevated ALP levels at the most recent time of progression were 5.7 (95% CI, 2.4-13.3) times more likely to have a liver metastases compared to patients with normal levels. Additionally, patients with elevated ALP levels at their most recent visit were 4.2 (95% CI, 1.7-10.7) times more likely to have a worse prognosis compared to patients with normal levels. However, after controlling for the effects of liver metastases, the association between elevated levels and prognosis was no longer significant. After controlling for the effects of age, sex, and liver metastases, large changes in AP levels were associated with a 4.4 (95% CI, 1.0-19.1) times greater odds of having a worse prognosis compared to patients with a minimal change. Patients with an ALP level greater than 160

  10. 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

  11. 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.

  12. 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.

  13. Glacial isostatic uplift of the European Alps

    NASA Astrophysics Data System (ADS)

    Mey, Jürgen; Scherler, Dirk; Wickert, Andrew D.; Egholm, David L.; Tesauro, Magdala; Schildgen, Taylor F.; Strecker, Manfred R.

    2016-11-01

    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.

  14. ALPASS: Teleseismic Tomography of the Eastern Alps

    NASA Astrophysics Data System (ADS)

    Brueckl, E.; Mitterbauer, U.; Lippitsch, R.; Behm, M.; ALPASS Working Group

    2007-12-01

    The Eastern Alps were formed by the north-south directed collision of the Adriatic (African) and European plates and a subsequent tectonic escape of crustal fragments to the unconstrained margin in the east, represented by the Pannonian Basin. Recent controlled source seismic experiments (TRANSALP, CELEBRATION 2000, and ALP 2002) revealed significant internal structures of the crust and the Moho topography. However, deeper plate tectonic structures (e.g. subducting slab) are still under debate. ALPASS is a passive seismic monitoring project aiming to reveal lower lithosphere and upper mantle beneath the wider Eastern Alpine region, and to contribute to a better understanding of the geodynamic processes at work. By cooperation of Austria, Croatia, Finland, Hungary, Poland, and USA 57 temporary seismic recording stations were deployed from May 2005 until May 2006. The layout was designed to extend the efforts of earlier experiments (e.g. TRANSALP) and to support two other passive seismic experiments (BOHEMA, Carpathian Basin Project), which are overlapping in the investigation area. Additionally, data from permanent networks was collected to improve coverage of the investigation area. 144 events (50% with M > 5.6) from epicentre distances between 30° and 100° were selected for teleseismic inversion. Travel time picking of P-wave arrivals has been done by a semi-automatic correlation technique. Crustal corrections benefit from the high resolution velocity model of the crust and the new Moho map derived from CELEBRATION 2000 and ALP 2002 data. First results of teleseismic inversion will be presented and discussed with respect to crustal structures revealed by the controlled source experiments, tomographic models generated during earlier studies, and their consequences for the conception of plate tectonics in the Eastern Alps.

  15. Sustained release of Semaphorin 3A from α-tricalcium phosphate based cement composite contributes to osteoblastic differentiation of MC3T3-E1 cells

    NASA Astrophysics Data System (ADS)

    Wang, Jin-Ning; Pi, Bin; Wang, Peng; Li, Xue-Feng; Yang, Hui-Lin; Zhu, Xue-Song

    2015-09-01

    The reinforcement of calcium phosphate materials with silk fibroin (SF) has been one of the strategies to overcome the brittleness. However, the lack of osteoinductivity may still restrict their further use. This study aimed to investigate the biocompatibility and osteogenesis capacity of a novel Semaphorin 3A-loaded chitosan microspheres/SF/α-tricalcium phosphate composite (Sema3A CMs/SF/α-TCP) in vitro. Sema3A was first incorporated into CMs, and the Sema3A CMs/SF/α-TCP composite was then prepared. The morphology of the CMs was observed using SEM. The in vitro release kinetics, cytotoxicity, and cell compatibility were evaluated, and the real-time quantitative polymerase chain reaction (RT-qPCR) and activity of alkaline phosphatase (ALP) were used to evaluate the osteogenesis capacity of the composite. The in vitro release of Sema3A from the Sema3A CMs/SF/α-TCP composite showed a temporally controlled manner. The extract of the Sema3A CMs/SF/α-TCP composite presented no obvious side effect on the MC3T3-E1 cell proliferation, nor promote cell proliferation. The MC3T3-E1 cells were well-spread and presented an elongated shape on the Sema3A CMs/SF/α-TCP composite surface; the ALP activity and the osteogenic-related gene expression were higher than those seeded on the surface of the CMs/SF/α-TCP and SF/α-TCP composites. In conclusion, Sema3A CMs/SF/α-TCP has excellent biocompatibility and contributes to the osteoblastic differentiation of MC3T3-E1 cells.

  16. Characterisation of rat and human tissue alkaline phosphatase isoforms by high-performance liquid chromatography and agarose gel electrophoresis.

    PubMed

    Dziedziejko, Violetta; Safranow, Krzysztof; Slowik-Zylka, Dorota; Machoy-Mokrzynska, Anna; Millo, Barbara; Machoy, Zygmunt; Chlubek, Dariusz

    2009-03-01

    Alkaline phosphatase (ALP) exists as several isoenzymes and many isoforms present in tissues and serum. The objective of this study was to separate tissue ALP forms in rats and humans and characterise their properties. The materials for the investigation were intestinal, bone, and liver tissue of rats and commercially available human preparations of tissue ALP. Two methods of separation were used: high-performance liquid chromatography (HPLC) and agarose gel electrophoresis. Using HPLC in the rat tissues, two ALP isoforms in the intestine, one in the bone, and three in the liver were identified. In humans three intestinal, two bone, and one liver isoform were resolved. Electrophoresis showed two ALP activity bands in rat intestine, one wide band in the bone, and three bands in the liver. ALP of human tissues was visualised as a single wide band, with a different mobility observed for each organ. In both species the presence of a form with properties characteristic of the bone isoform of the tissue-nonspecific isoenzyme was observed in the intestine. HPLC offers a higher resolution than electrophoresis with respect to tissue ALP fractions in rats and in humans, but electrophoresis visualises high-molecular-mass insoluble enzyme forms.

  17. Alkaline phosphatase determines polyphosphate-induced mineralization in a cell-type independent manner.

    PubMed

    Mikami, Yoshikazu; Tsuda, Hiromasa; Akiyama, Yuko; Honda, Masaki; Shimizu, Noriyoshi; Suzuki, Naoto; Komiyama, Kazuo

    2016-11-01

    Polyphosphate [Poly(P)] has positive effects on osteoblast mineralization; however, the underlying mechanism remains unclear. In addition, it is unknown whether Poly(P) promotes mineralization in soft tissues. We investigated this by using various cells. Poly(P) concentrations of 1 and 0.5 mg/mL yielded high levels of mineralization in ROS17/2.8 osteoblast cells. Similarly, Poly(P) induced mineralization in cell types expressing alkaline phosphatase (ALP), namely, ATDC5 and MC3T3-E1, but not in CHO, C3H10T1/2, C2C12, and 3T3-L1 cells. Furthermore, forced expression of ALP caused Poly(P)-induced mineralization in CHO cells. These results suggest that ALP determines Poly(P)-induced mineralization in a cell-type independent manner.

  18. A smart fluorescence nanoprobe for the detection of cellular alkaline phosphatase activity and early osteogenic differentiation.

    PubMed

    Cao, Feng-Yi; Fan, Jin-Xuan; Long, Yue; Zeng, Xuan; Zhang, Xian-Zheng

    2016-07-01

    In the past decades, biomaterials were designed to induce stem cell toward osteogenic differentiation. However, conventional methods for evaluation osteogenic differentiation all required a process of cell fixation or lysis, which induce waste of a large number of cells. In this study, a fluorescence nanoprobe was synthesized by combining phosphorylated fluoresceinamine isomer I (FLA) on the surface of mesoporous silica-coated superparamagnetic iron oxide (Fe3O4@mSiO2) nanoparticles. In the presence of alkaline phosphatase (ALP), the phosphorylated FLA on the nanoprobe would be hydrolyzed, resulting in a fluorescence recovery of FLA. During early osteogenic differentiation, a high-level expression of cellular ALP was induced, which accelerated the hydrolysis of phosphorylated FLA, resulting in an enhancement of cellular fluorescence intensity. This fluorescence nanoprobe provides us a rapid and non-toxic method for the detection of cellular ALP activity and early osteogenic differentiation.

  19. The measurement of alkaline phosphatase at nanomolar concentration within 70 s using a disposable microelectrochemical transistor.

    PubMed

    Astier, Y; Bartlett, P N

    2004-08-01

    We report a new approach to the measurement of alkaline phosphatase concentration based on the use of a disposable poly(aniline) microelectrochemical transistor. The measurement is carried out in a two cell configuration in which the poly(aniline) microelectrochemical transistor operates in acid solution and is connected to the alkaline buffer solution containing the analyte by a salt bridge. Disposable microelectrochemical transistors were reproducibly fabricated by electrochemical deposition of poly(aniline) onto photolithographically fabricated gold microband arrays. Using these devices alkaline phosphatase was detected by employing p-aminophenyl phosphate as the substrate for the enzyme and using glucose and glucose oxidase to recycle the p-aminophenol generated upon enzyme catalysed hydrolysis of the phosphate. Recycling the p-aminophenol with glucose and glucose oxidase amplified the detection of alkaline phosphatase approximately tenfold. Using this approach we obtain linear calibration curves for alkaline phosphatase up to 5 nM within 70 s on single use devices.

  20. Secreted Aspergillus fumigatus Protease Alp1 Degrades Human Complement Proteins C3, C4, and C5▿

    PubMed Central

    Behnsen, Judith; Lessing, Franziska; Schindler, Susann; Wartenberg, Dirk; Jacobsen, Ilse D.; Thoen, Marcel; Zipfel, Peter F.; Brakhage, Axel A.

    2010-01-01

    The opportunistic human pathogenic fungus Aspergillus fumigatus is a major cause of fungal infections in immunocompromised patients. Innate immunity plays an important role in the defense against infections. The complement system represents an essential part of the innate immune system. This cascade system is activated on the surface of A. fumigatus conidia and hyphae and enhances phagocytosis of conidia. A. fumigatus conidia but not hyphae bind to their surface host complement regulators factor H, FHL-1, and CFHR1, which control complement activation. Here, we show that A. fumigatus hyphae possess an additional endogenous activity to control complement activation. A. fumigatus culture supernatant efficiently cleaved complement components C3, C4, C5, and C1q as well as immunoglobulin G. Secretome analysis and protease inhibitor studies identified the secreted alkaline protease Alp1, which is present in large amounts in the culture supernatant, as the central molecule responsible for this cleavage. An alp1 deletion strain was generated, and the culture supernatant possessed minimal complement-degrading activity. Moreover, protein extract derived from an Escherichia coli strain overproducing Alp1 cleaved C3b, C4b, and C5. Thus, the protease Alp1 is responsible for the observed cleavage and degrades a broad range of different substrates. In summary, we identified a novel mechanism in A. fumigatus that contributes to evasion from the host complement attack. PMID:20498262

  1. [Effect of phosphate upon calcium intestinal absorption (author's transl)].

    PubMed

    Fournier, P; Dupuis, Y; Digaud, A; Fournier, A

    1976-11-01

    Adult rats receive 5 to 50 mM CaCl2 solutions in which glycerophosphate or sodium diacid phosphate may be added in variable quantity. These solutions are administered by gavage or in situ ligatured jejunal loop. The inhibition of calcium absorption dependent on simultaneously administered phosphate doses is well characterized: high for the lowest concentration, the inhibiting effect of phosphate doses decreases more and more reaching a limit from which phosphate supplementation has no effect. These observations discarding an intervention of phosphate by calcium insolubilization seem to demonstrate that the control supplied by phosphates on calcium absorption is of enzymatic character. Facts related to the respective effects of calcium and phosphates on the action of alkaline phosphatases lead to discuss a possible intervention of these enzymes upon calcium transfer.

  2. Fabrication of Poly-l-lactic Acid/Dicalcium Phosphate Dihydrate Composite Scaffolds with High Mechanical Strength-Implications for Bone Tissue Engineering.

    PubMed

    Tanataweethum, Nida; Liu, Wai Ching; Goebel, W Scott; Li, Ding; Chu, Tien Min

    2015-11-04

    Scaffolds were fabricated from poly-l-lactic acid (PLLA)/dicalcium phosphate dihydrate (DCPD) composite by indirect casting. Sodium citrate and PLLA were used to improve the mechanical properties of the DCPD scaffolds. The resulting PLLA/DCPD composite scaffold had increased diametral tensile strength and fracture energy when compared to DCPD only scaffolds (1.05 vs. 2.70 MPa and 2.53 vs. 12.67 N-mm, respectively). Sodium citrate alone accelerated the degradation rate by 1.5 times independent of PLLA. Cytocompatibility of all samples were evaluated using proliferation and differentiation parameters of dog-bone marrow stromal cells (dog-BMSCs). The results showed that viable dog-BMSCs attached well on both DCPD and PLLA/DCPD composite surfaces. In both DCPD and PLLA/DCPD conditioned medium, dog-BMSCs proliferated well and expressed alkaline phosphatase (ALP) activity indicating cell differentiation. These findings indicate that incorporating both sodium citrate and PLLA could effectively improve mechanical strength and biocompatibility without increasing the degradation time of calcium phosphate cement scaffolds for bone tissue engineering purposes.

  3. Fabrication of Poly-l-lactic Acid/Dicalcium Phosphate Dihydrate Composite Scaffolds with High Mechanical Strength—Implications for Bone Tissue Engineering

    PubMed Central

    Tanataweethum, Nida; Liu, Wai Ching; Scott Goebel, W.; Li, Ding; Chu, Tien Min

    2015-01-01

    Scaffolds were fabricated from poly-l-lactic acid (PLLA)/dicalcium phosphate dihydrate (DCPD) composite by indirect casting. Sodium citrate and PLLA were used to improve the mechanical properties of the DCPD scaffolds. The resulting PLLA/DCPD composite scaffold had increased diametral tensile strength and fracture energy when compared to DCPD only scaffolds (1.05 vs. 2.70 MPa and 2.53 vs. 12.67 N-mm, respectively). Sodium citrate alone accelerated the degradation rate by 1.5 times independent of PLLA. Cytocompatibility of all samples were evaluated using proliferation and differentiation parameters of dog-bone marrow stromal cells (dog-BMSCs). The results showed that viable dog-BMSCs attached well on both DCPD and PLLA/DCPD composite surfaces. In both DCPD and PLLA/DCPD conditioned medium, dog-BMSCs proliferated well and expressed alkaline phosphatase (ALP) activity indicating cell differentiation. These findings indicate that incorporating both sodium citrate and PLLA could effectively improve mechanical strength and biocompatibility without increasing the degradation time of calcium phosphate cement scaffolds for bone tissue engineering purposes. PMID:26556380

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

    PubMed Central

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

    2017-01-01

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

  5. A ratiometric fluorescent probe for alkaline phosphatase via regulation of excited-state intramolecular proton transfer.

    PubMed

    Fan, Chunlei; Luo, Shengxu; Qi, Haiping

    2016-03-01

    A ratiometric fluorescent probe 2-(benzimidazol-2-yl)phenyl phosphoric acid (1) for alkaline phosphatase (ALP) is designed and synthesized. The method employs the modulation of the excited-state intramolecular proton transfer (ESIPT) process of 2-(2'-hydroxyphenyl)benzimidazole (HPBI) through the hydroxyl group protection/deprotection reaction. Upon phosphorylated with POCl3 , HPBI shows only an emission peak at 363 nm due to the blockage of ESIPT. However, once selective enzymatic hydrolysis with alkaline phosphatase (ALP) in Tris-HCl buffer occurs, the probe 1 is returned to HPBI and the ESIPT process is switched on, which results in a decrease in the emission band at 363 nm and an increase in a new fluorescence peak around 430 nm. The fluorescence intensity ratio at 430 and 360 nm (I430/I360) increases linearly with the activity of ALP up to 0.050 U/mL and the detection limit is 0.0013 U/mL. The proposed probe shows excellent specificity toward ALP.

  6. Marrow stromal osteoblast function on a poly(propylene fumarate)/beta-tricalcium phosphate biodegradable orthopaedic composite.

    PubMed

    Peter, S J; Lu, L; Kim, D J; Mikos, A G

    2000-06-01

    The objective of this study was to assess the osteoconductivity of a poly(propylene fumarate)/beta-tricalcium phosphate (PPF/beta-TCP) composite in vitro. We examined whether primary rat marrow stromal cells would attach, proliferate, and express differentiated osteoblastic function when seeded on PPF/beta-TCP substrates. Attachment studies showed that a confluent monolayer of cells had adhered to the substrates within an 8 h time frame for marrow stromal cells seeded at confluent numbers. Proliferation and differentiated function of the cells were then investigated for a period of 4 weeks for an initial seeding density of 42,000 cells/cm2. Rapid proliferation during the first 24 h as determined by 3H-thymidine incorporation was mirrored by an initial rapid increase in total cell number by DNA assay. A lower proliferation rate and a gradual increase in cell number persisted for the remainder of the study, resulting in a final cell number of 128,000 cells/cm2. Differentiated cell function was assessed by measuring alkaline phosphatase (ALP) activity and osteocalcin (OC) production throughout the time course. Both markers of osteoblastic differentiation increased significantly over a 4-week period. By day 28, cells grown on PPF/beta-TCP reached a maximal ALP activity of 11 (+/- 1) x 10(-7) micromol/min/cell, while the OC production reached 40 (+/- 1) x 10(-6) ng/cell. These data show that a PPF/beta-TCP composite exhibits in vitro osteoconductivity similar to or better than that of control tissue culture polystyrene.

  7. Alkaline flooding injection strategy

    SciTech Connect

    French, T.R.; Josephson, C.B.

    1992-03-01

    The objective of this project is to improved alkali-surfactant flooding methods, and this includes determining the proper design of injection strategy. Several different injection strategies have been used or suggested for recovering heavy oils with surfactant-enhanced alkaline flooding methods. Oil recovery was compared for four different injection strategies: (1) surfactant followed by polymer, (2) surfactant followed by alkaline polymer, (3) alkaline surfactant followed by polymer, and (4) alkali, surfactant, and polymer mixed in a single formulation. The effect of alkaline preflush was also studied under two different conditions. All of the oil recovery experiments were conducted under optimal conditions with a viscous, non-acidic oil from Hepler (KS) oil field. The coreflood experiments were conducted with Berea sandstone cores since field core was not available in sufficient quantity for coreflood tests. The Tucker sand of Hepler field is a Class I fluvial dominated deltaic reservoir, as classified by the Department of Energy, which has been selected as the site of a DOE-sponsored field pilot test.

  8. Ratiometric fluorescent response of electrospun fibrous strips for real-time sensing of alkaline phosphatase in serum.

    PubMed

    Zhao, Long; Xie, Songzhi; Song, Xiaojie; Wei, Jiaojun; Zhang, Zhao; Li, Xiaohong

    2017-05-15

    The development of rapid, convenient and reliable assays for monitoring alkaline phosphatase (ALP) levels is valuable for clinical diagnoses and biomedical research. In the current study, a ratiometric assay of ALP activity has been realized by covalent immobilization of fluorescein onto polyethylene terephthalate (PET) fibers, followed by electrostatic adsorption of bisquaternary ammonium salt of tetraphenylethene (TPE-2N(+)). In the absence of ALP, the complex formation between phosphorylated fluorescein and TPE-2N(+) results in the aggregation-induced emission (AIE) of TPE at 471nm. While in the presence of ALP, the hydrolysis of phosphoesters leads to a gradual removal of TPE-2N(+) and the restoration of fluorescein emission at 514nm. Fibers with surface amine densities of 30 nmol/mg show the most significant and almost linear increases in I514/I471 ratios from 0.73 to 3.05 with increasing ALP concentrations from 0 to 100 mU/mL. The ratiometric fluorescence responses result in color changes of fibrous strips from blue (TPE-2N(+)) to green (fluorescein) under an ultraviolet lamp in a matter of minutes. The color changes are more suitable for an eyeball detection of ALP levels ranging from 0 to 80 mU/mL, which is included in the concentration range of ALP in human serum considering the dilution factor if necessary. The ALP detection indicates no apparent interference by serum components and good agreement with enzyme-linked immunosorbent assay (ELISA). Thus, this is the first study on ratiometric fluorescent assay of serum ALP levels by fibrous strips, which offers a capacity to exploit electrospun fibrous mats and ratiometric responses for real-time assays of bioactive substances as self-test devices.

  9. Pre-treatment serum alkaline phosphatase and lactate dehydrogenase as prognostic factors in triple negative breast cancer

    PubMed Central

    Chen, Bo; Dai, Danian; Tang, Hailin; Chen, Xi; Ai, Xiaohong; Huang, Xiaojia; Wei, Weidong; Xie, Xiaoming

    2016-01-01

    Background: Serum parameters as prognostic parameters are studied widely. We aim to examine the prognostic significance of the serum alkaline phosphatase (ALP) level and lactate dehydrogenase (LDH) level in triple negative breast cancer (TNBC). Methods: Total of 253 TNBC patients from Sun Yat-sen University Cancer Center who underwent treatment between January 2004 and December 2009 was conducted in this retrospective study. Before treatment serum ALP and LDH levels were routinely measured. We use the receiver operating characteristic (ROC) curve analysis to estimate the cutoff value of serum ALP and LDH levels. The Kaplan-Meier method and multivariable Cox regression analysis were used for Disease free survival (DFS) and overall survival (OS) assessed. Results: The ROC curves determined that the optimum cutoff point for ALP and LDH were 66.5u/L and 160.5u/L, respectively. The elevated ALP and LDH were both significantly associated with decreased DFS and OS (both P < 0.001). In addition, the entire cohort was stratified into three subgroups basis of ALP levels and LDH levels. TNBC Patients who with ALP >66.5 u/L and LDH >160.5u/L had the worst DFS and OS (both P < 0.001). In TNBC patients, univariate and multivariate Cox regression analyses conformed ALP and LDH were independent unfavorable prognostic factors for DFS and OS. Conclusions: The serum levels of ALP and LDH before treatment are independent prognostic parameters and may serve as complement to help predict survival in TNBC. PMID:27994669

  10. Effects of macromolecular crowding on alkaline phosphatase unfolding, conformation and stability.

    PubMed

    Jia, Jiajia; Peng, Xin; Qi, Wei; Su, Rongxin; He, Zhimin

    2017-03-23

    The interior of the cell is tightly packed with various biological macromolecules, which affects physiological processes, especially protein folding process. To explore how macromolecular crowding may influence protein folding process, alkaline phosphatase (ALP) was chosen as a model protein, and the unfolding process of ALP induced by GdnHCl was studied in the presence of crowding agents such as PEG 4000, Dextran 70 and Ficoll 70. The effect of macromolecular crowding on the denatured state of ALP was directly probed by measuring enzyme activities, fluorescence spectroscopy and circular dichroism. From the results of circular dichroism, GdnHCl induced a biphasic change, suggesting that a three-state unfolding mechanism was involved in the denaturation process irrespective of the absence or presence of crowding agents. It was also found that crowding agents had a little impact on the unfolding process of ALP. The results of phase diagrams also demonstrated that the unfolding process of ALP induced by GdnHCl was three-state mechanism. Moreover, the results of fluorescence spectra demonstrated that with the increase of GdnHCl concentration, the structure of protein had changed, but existence of crowding agents can make protein structure more stable. Our results can provide valuable information for understanding the protein folding in vivo.

  11. Bone-specific alkaline phosphatase activity is inhibited by bisphosphonates: role of divalent cations.

    PubMed

    Vaisman, Diego N; McCarthy, Antonio D; Cortizo, Ana M

    2005-05-01

    Bisphosphonates (BPs) are drugs widely used in the treatment of various bone diseases. BPs localize to bone mineral, and their concentration in resorption lacunae could reach almost millimolar levels. Bone alkaline phosphatase (ALP) is a membrane-bound exoenzyme that has been implicated in bone formation and mineralization. In this study, we investigated the possible direct effect of three N-containing BPs (alendronate, pamidronate, and zoledronate) on the specific activity of bone ALP obtained from an extract of UMR106 rat osteosarcoma cells. Enzymatic activity was measured by spectrophotometric detection of p-nitrophenol product and by in situ visualization of ALP bands after an electrophoresis on cellulose acetate gels. Because ALP is a metalloprotein that contains Zn2+ and Mg2+, both of which are necessary for catalytic function, we also evaluated the participation of these divalent cations in the possible effect of BPs on enzymatic activity. All BPs tested were found to dose-dependently inhibit spectrophotometrically measured ALP activity (93-42% of basal) at concentrations of BPs between 10-5 M and 10-4 M, the order of potency being zoledronate approximately equals alendronate > pamidronate. However, coincubation with excess Zn2+ or Mg2+ completely abolished this inhibitory effect. Electrophoretic analysis rendered very similar results: namely a decrease in the enzymatic activity of the bone-ALP band by BPs and a reversion of this inhibition by divalent cations. This study shows that N-containing BPs directly inhibit bone-ALP activity, in a concentration range to which this exoenzyme is probably exposed in vivo. In addition, this inhibitory effect is most possibly the result of the chelation of Zn2+ and Mg2+ ions by BPs.

  12. Characteristics of plasmalemma alkaline phosphatase of rat mesenteric artery.

    PubMed

    Kwan, C Y

    1983-01-01

    General characteristics of alkaline phosphatase activity of the plasma membrane-enriched fraction isolated from rat mesenteric arteries were investigated. The vascular smooth muscle plasmalemma alkaline phosphatase is a metalloenzyme which is strongly inhibited by chelating agents and this inhibition can be completely overcome by addition of Mg2+ or Ca2+. Zn2+ only partially reactivates the enzyme in the presence of low concentrations of EDTA. The enzymatic hydrolysis of p-nitrophenyl phosphate, beta-glycerophosphate, alpha-glycerophosphate, or 3'-adenosine monophosphate showed an optimal activity in the alkaline region between pH 9 and 11. The alkaline phosphatase activity is distinctly different from the plasmalemma ATPase and 5'-nucleotidase activities with respect to their pH dependence, influence by added divalent metal ions and stability against heat inactivation. Vanadate ion, being structurally similar to the transition state analog of the phosphoryl group, potently inhibits alkaline phosphatase with an apparent Ki of 1.5 microM. The altered alkaline phosphatase activity of vascular smooth muscle in relation to its possible physiological function and pathophysiological manifestation associated with hypertensive disease are discussed.

  13. Alkaline quinone flow battery.

    PubMed

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

    2015-09-25

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

  14. Advanced alkaline water electrolysis

    NASA Astrophysics Data System (ADS)

    Wakabayashi, N.; Torikai, E.; Kawami, Y.; Takenaka, H.

    Results are presented of experimental studies of possible separators and electrodes for use in advanced, high-temperature, high-pressure alkaline water electrolyzers. Material evaluations in alkaline water electrolyzers at temperatures from 100 to 120 C have shown a new type polytetrafluoroethylene membrane impregnated with potassium titanate to be the most promising when the separator is prepared by the hydrothermal treatment of a porous PFTE membrane impregnated with hydrated titanium oxide. Measurements of cell voltages in 30% KOH at current densities from 5 to 100 A/sq dm at temperatures up to 120 C with nickel electrodes of various structures have shown the foamed nickel electrode, with an average pore size of 1-1.5 mm, to have the best performance. When the foamed nickel is coated by fine powdered nickel, carbonyl nickel or Raney nickel to increase electrode surface areas, even lower cell voltages were found, indicating better performance.

  15. 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.

  16. 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,…

  17. Prevalence of acute mountain sickness in the Eastern Alps.

    PubMed

    Mairer, Klemens; Wille, Maria; Bucher, Thomas; Burtscher, Martin

    2009-01-01

    Little information is available on the prevalence of acute mountain sickness (AMS) in the Eastern Alps compared with the Western Alps. Because of differences regarding the populations of mountaineers, we hypothesized that the prevalence differs between the Eastern and Western Alps. Thus, we determined the prevalence and risk factors of AMS at four different altitudes in the Eastern Alps of Austria. Four hundred and thirty-one recreational hikers were studied using questionnaires on the morning of their first night at high altitude. A diagnosis of AMS was based on a Lake Louise Score > or =4, the presence of headache, and at least one additional symptom. Overall 16.2% of the subjects met the criteria for AMS, and the prevalence of AMS increased significantly with altitude (2200 m: 6.9%; 2500 m: 9.1%; 2800 m: 17.4%; 3500 m: 38.0%). Heavy perceived exertion, a history of migraine, the absolute altitude reached, little mountaineering experience, and inadequate water intake (< or =2 L) were independent AMS risk factors. The reported altitude-related AMS prevalence in the Western Alps is 4% to 8% lower compared with that found in this study for the Eastern Alps. In conclusion, the prevalence of AMS is higher in the tourist population of the Eastern Alps compared to the more experienced mountaineers of the Western Alps. Consideration of easily modifiable risk factors such as individual exertion and water intake could markedly reduce AMS and contribute to the enjoyment of mountaineering.

  18. 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.

  19. Biocompatibility and osteogenesis of calcium phosphate composite scaffolds containing simvastatin-loaded PLGA microspheres for bone tissue engineering.

    PubMed

    Zhang, Hao-Xuan; Xiao, Gui-Yong; Wang, Xia; Dong, Zhao-Gang; Ma, Zhi-Yong; Li, Lei; Li, Yu-Hua; Pan, Xin; Nie, Lin

    2015-10-01

    By utilizing a modified solid/oil/water (s/o/w) emulsion solvent evaporation technique, calcium phosphate composite scaffolds containing simvastatin-loaded PLGA microspheres (SIM-PLGA-CPC) were prepared in this study. We characterized the morphology, encapsulation efficiency and in vitro drug release of SIM-loaded PLGA microspheres as well as the macrostructure, pore size, porosity and mechanical strength of the scaffolds. Rabbit bone mesenchymal stem cells (BMSCs) were seeded onto SIM-PLGA-CPC scaffolds, and the proliferation, morphology, cell cycle and differentiation of BMSCs were investigated using the cell counting kit-8 (CCK-8) assay, scanning electron microscopy (SEM), flow cytometry, alkaline phosphatase (ALP) activity and alizarin red S staining, respectively. The results revealed that SIM-PLGA-CPC scaffolds were biocompatible and osteogenic in vitro. To determine the in vivo biocompatibility and osteogenesis of the scaffolds, both pure PLGA-CPC scaffolds and SIM-PLGA-CPC scaffolds were implanted in rabbit femoral condyles and microradiographically and histologically investigated. SIM-PLGA-CPC scaffolds exhibited good biocompatibility and could improve the efficiency of new bone formation. All these results suggested that the SIM-PLGA-CPC scaffolds fulfilled the basic requirements of bone tissue engineering scaffold and possessed application potentials in orthopedic surgery.

  20. Transition-state structures for enzymatic and alkaline phosphotriester hydrolysis

    SciTech Connect

    Caldwell, S.R.; Raushel, F.M. ); Weiss, P.M.; Cleland, W.W. )

    1991-07-30

    The primary and secondary {sup 18}O isotope effects for the alkaline (KOH) and enzymatic (phosphotriesterase) hydrolysis of two phosphotriesters, O,O-diethyl p-nitrophenyl phosphate (I) and O,O-diethyl O-(4-carbamoylphenyl) phosphate (II), are consistent with an associative mechanism with significant changes in bond order to both the phosphoryl and phenolic leaving group oxygens in the transition state. The synthesis of ({sup 15}N, phosphoryl-{sup 18}O)-,({sup 15}N, phenolic-{sup 18}O)-, and ({sup 15}N)-O,O-diethyl p-nitrophenyl phosphate and O,O-diethyl O-(4-carbamoylphenyl)phosphate is described. The primary and secondary {sup 18}O isotope effects for the alkaline hydrolysis of compound I are 1.0060 and 1.0063 {plus minus} 0.0001, whereas for compound II they are 1.027{plus minus}0.002 and 1.025 {plus minus} 0.002, respectively. These isotope effects are consistent with the rate-limiting addition of hydroxide and provide evidence for a S{sub N}2-like transition state with the absence of a stable phosphorane intermediate. For the enzymatic hydrolysis of compound I, the primary and secondary {sup 18}O isotope effects are very small, 1.0020 and 1.0021{plus minus}0.0004, respectively, and indicate that the chemical step in the enzymatic mechanism is not rate-limiting. The {sup 18}O isotope effects for the enzymatic hydrolysis of compound II are 1.036{plus minus}0.001 and 1.0181{plus minus}0.0007, respectively, and are comparable in magnitude to the isotope effects for alkaline hydrolysis, suggesting that the chemical step is rate-limiting. The relative magnitude of the primary {sup 18}O isotope effects for the alkaline and enzymatic hydrolysis of compound II reflect a transition state that is more progressed for the enzymatic reaction.

  1. Transition-state structures for enzymatic and alkaline phosphotriester hydrolysis.

    PubMed

    Caldwell, S R; Raushel, F M; Weiss, P M; Cleland, W W

    1991-07-30

    The primary and secondary 18O isotope effects for the alkaline (KOH) and enzymatic (phosphotriesterase) hydrolysis of two phosphotriesters, O,O-diethyl p-nitrophenyl phosphate (I) and O,O-diethyl O-(4-carbamoylphenyl) phosphate (II), are consistent with an associative mechanism with significant changes in bond order to both the phosphoryl and phenolic leaving group oxygens in the transition state. The synthesis of [15N, phosphoryl-18O]-, [15N, phenolic-18O]-, and [15N]-O,O-diethyl p-nitrophenyl phosphate and O,O-diethyl O-(4-carbamoylphenyl)phosphate is described. The primary and secondary 18O isotope effects for the alkaline hydrolysis of compound I are 1.0060 and 1.0063 +/- 0.0001, whereas for compound II they are 1.027 +/- 0.002 and 1.025 +/- 0.002, respectively. These isotope effects are consistent with the rate-limiting addition of hydroxide and provide evidence for a SN2-like transition state with the absence of a stable phosphorane intermediate. For the enzymatic hydrolysis of compound I, the primary and secondary 18O isotope effects are very small, 1.0020 and 1.0021 +/- 0.0004, respectively, and indicate that the chemical step in the enzymatic mechanism is not rate-limiting. The 18O isotope effects for the enzymatic hydrolysis of compound II are 1.036 +/- 0.001 and 1.0181 +/- 0.0007, respectively, and are comparable in magnitude to the isotope effects for alkaline hydrolysis, suggesting that the chemical step is rate-limiting. The relative magnitude of the primary 18O isotope effects for the alkaline and enzymatic hydrolysis of compound II reflect a transition state that is more progressed for the enzymatic reaction.

  2. 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.

  3. Dephosphorylation of purine mononucleotides by alkaline phosphatases. Substrate specificity and inhibition patterns.

    PubMed

    Jensen, M H

    1979-11-09

    Three purine mononucleotides, adenosine-, inosine- and guanosine monophosphate, were used as substrates at pH 7.4 and at 10.4 for three alkaline phosphatases (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.1) containing similar phosphate-binding serine groups at their esteratic sites. Substrate specificity was found for the enzymes from calf intestine and bovine liver. Alkaline phosphatase from Escherichia coli was nonspecific. A substrate-dependent and pronounced inhibition with the purine analogue 1,3-dimethyl xanthine was found for the enzymes from intestine and liver, but not for alkaline phosphatase from E. coli. A substrate-independent and pronounced inhibition was found for all three enzymes with the phosphomonoester p-nitrophenol phosphate as the inhibitor. Alkaline phosphatases may play an important role in the regulation of the intracellular content of purine mononucleotides.

  4. Alp Transit: Crossing Faults 44 and 49

    NASA Astrophysics Data System (ADS)

    El Tani, M.; Bremen, R.

    2014-05-01

    This paper describes the crossing of faults 44 and 49 when constructing the 57 km Gotthard base tunnel of the Alp Transit project. Fault 44 is a permeable fault that triggered significant surface deformations 1,400 m above the tunnel when it was reached by the advancing excavation. The fault runs parallel to the downstream face of the Nalps arch dam. Significant deformations were measured at the dam crown. Fault 49 is sub-vertical and permeable, and runs parallel at the upstream face of the dam. It was necessary to assess the risk when crossing fault 49, as a limit was put on the acceptable dam deformation for structural safety. The simulation model, forecasts and action decided when crossing over the faults are presented, with a brief description of the tunnel, the dam, and the monitoring system.

  5. Glacial isostatic uplift of the European Alps

    NASA Astrophysics Data System (ADS)

    Mey, Juergen; Scherler, Dirk; Wickert, Andrew D.; Egholm, David L.; Tesauro, Magdala; Schildgen, Taylor F.; Strecker, Manfred R.

    2016-04-01

    Present-day vertical movements of the Earth's surface are mostly due to tectonic deformation, volcanic processes, and crustal loading/unloading. In tectonically stable regions of North America and Scandinavia, vertical movements are almost entirely attributable to glacial isostatic rebound after the melting of the Laurentide and Fennoscandian ice sheets. In contrast, the Pleistocene Alpine icecap grew on a younger mountain belt that formed by collision of the European and African plates, still subject to shortening. Therefore, measured uplift is potentially a composite signal of tectonic shortening and unloading after deglaciation and concomitant erosion. Deciphering the contributions of tectonics and crustal unloading to present-day uplift rates in formerly-glaciated mountain belts is a prerequisite to using uplift data to estimate the viscosity structure of the Earth's mantle, a key variable in geodynamics. We evaluate the post-LGM glacial-isostatic rebound of the Alps following a 4-tiered procedure. First, we estimated the thickness distribution of sedimentary valley fills to create a bedrock map of the entire mountain belt. Second, this map was used as topographic basis for the reconstruction of the Alpine icecap using a numerical ice-flow model. Third, we estimated the equilibrium deflection of the Alpine lithosphere, using the combined loads of ice and sediments with a variable effective elastic thickness. Finally, we used an exponential decay function to infer the residual deflection and the present-day uplift rate for a range of upper mantle viscosities. Our analysis shows that virtually all of the geodetically measured surface uplift in the Swiss and the Austrian Alps can be attributed to glacial unloading and redistribution of sediments, assuming an upper-mantle viscosity lower than that inferred for an old craton (e.g., Fennoscandia), but higher than that for a region with recent crustal thinning (e.g., Basin and Range province).

  6. Rapamycin improves lymphoproliferative disease in murine autoimmune lymphoproliferative syndrome (ALPS).

    PubMed

    Teachey, David T; Obzut, Dana A; Axsom, Kelly; Choi, John K; Goldsmith, Kelly C; Hall, Junior; Hulitt, Jessica; Manno, Catherine S; Maris, John M; Rhodin, Nicholas; Sullivan, Kathleen E; Brown, Valerie I; Grupp, Stephan A

    2006-09-15

    Autoimmune lymphoproliferative syndrome (ALPS) is a disorder of abnormal lymphocyte survival caused by defective Fas-mediated apoptosis, leading to lymphadenopathy, hepatosplenomegaly, and an increased number of double-negative T cells (DNTs). Treatment options for patients with ALPS are limited. Rapamycin has been shown to induce apoptosis in normal and malignant lymphocytes. Since ALPS is caused by defective lymphocyte apoptosis, we hypothesized that rapamycin would be effective in treating ALPS. We tested this hypothesis using rapamycin in murine models of ALPS. We followed treatment response with serial assessment of DNTs by flow cytometry in blood and lymphoid tissue, by serial monitoring of lymph node and spleen size with ultrasonography, and by enzyme-linked immunosorbent assay (ELISA) for anti-double-stranded DNA (dsDNA) antibodies. Three-dimensional ultrasound measurements in the mice correlated to actual tissue measurements at death (r = .9648). We found a dramatic and statistically significant decrease in DNTs, lymphadenopathy, splenomegaly, and autoantibodies after only 4 weeks when comparing rapamycin-treated mice with controls. Rapamycin induced apoptosis through the intrinsic mitochondrial pathway. We compared rapamycin to mycophenolate mofetil, a second-line agent used to treat ALPS, and found rapamycin's control of lymphoproliferation was superior. We conclude that rapamycin is an effective treatment for murine ALPS and should be explored as treatment for affected humans.

  7. An association between time-varying serum alkaline phosphatase concentrations and mortality rate in patients undergoing peritoneal dialysis: a five-year cohort study

    PubMed Central

    Liu, Ying; Zhu, Jin-Gang; Cheng, Ben-Chung; Liao, Shang-Chih; Lee, Chih-Hsiung; Chang, Wen Xiu; Chen, Jin-Bor

    2017-01-01

    The relationship between serum alkaline phosphatase (ALP) concentrations and mortality in peritoneal dialysis (PD) patients is rarely reported. We enrolled 667 PD patients in one PD centre in Taiwan to retrospectively examine the association between three ALP concentrations (baseline, time-averaged, time-dependent) and mortality over a 5-year period (2011–2015). Baseline data collection included demographics, clinical, and laboratory parameters. Multivariable-adjusted Cox models were used to analyse the association. Four ALP quartiles were defined at the baseline: ≤62, 63–82, 83–118, and ≥119 U/L. Of 667 patients, 65 patients died, of which 8 patients died due to cardiovascular disease. Females were predominant in the higher ALP quartiles, and 24-h urine volume was significantly proportionately decreased in the higher ALP quartiles. ALP quartiles expressed by the three models were not associated with all-cause or cardiovascular mortalities after adjusting for demographics, liver function, bone metabolism, mortality, hemoglobin, and 24-h urine volume. In conclusion, ALP concentrations were not associated with death risk in PD patients over the 5-year period. PMID:28256582

  8. Trace elements in metapelitic schists from Alpe Sponda (Central Alps, Switzerland)

    NASA Astrophysics Data System (ADS)

    Redler, Charlotte; Irouschek, Alfred; Jeffries, Teresa; Gieré, Reto

    2015-04-01

    Lenses of kyanite-cordierite-garnet-bearing paragonite-biotite schist were found in paragonite-free metapelitic rocks at Pizzo Forno, near Alpe Sponda (Simano Nappe, Central Alps). These lenses are mineralogically distinct, as they lack both quartz and staurolite, but they contain large amounts of tourmaline (up to 20-25 vol.%). Tourmaline shows strong colour zoning, which in combination with the major and trace element data indicates at least two stages of crystal growth during prograde metamorphism. Mineral equilibrium modelling based on the bulk-rock compositions yielded peak metamorphic conditions of amphibolite-facies grade (T = 590-660 °C and P = 6.5-7.8 kbar), which is consistent with the P-T conditions estimated previously for this part of the Central Alps. The light elements lithium and boron are strongly enriched in the Alpe Sponda samples, which therefore show unusual bulk-rock concentrations. Lithium is mainly distributed to cordierite (450 ppm) and to a lower extent to muscovite (250 ppm), biotite (180 ppm) and paragonite; all other minerals show only minor amounts of lithium. As commonly expected, boron is partitioned into tourmaline and to a much lesser amount into muscovite, whereas all other minerals have low boron contents. In addition, cordierite has a relatively high Be content, underlining the hypotheses of the presence of Na-Be cordierite. The analysis of rare earth elements (REE) shows that all samples are enriched in low REE (LREE; La-Eu) relative to the heavy REE (HREE: Gd-Lu), and have a small negative Ce anomaly, as well as a pronounced negative Eu anomaly, which can be illustrated in a chondrite-normalised diagram. Furthermore, all minerals that do not show a distinct positive Eu anomaly (i.e. garnet, biotite, muscovite, cordierite, kyanite, chlorite and rutile) show a marked negative Ce anomaly. In addition, plagioclase and tourmaline exhibit a pronounced positive Eu anomaly but no Ce anomaly. Especially in the case of tourmaline the

  9. Effect of low level laser therapy and zoledronate on the viability and ALP activity of Saos-2 cells.

    PubMed

    Bayram, Hilal; Kenar, Halime; Taşar, Ferda; Hasırcı, Vasıf

    2013-01-01

    A limited number of clinical studies indicate the supportive role of low level laser therapy (LLLT) on medical and/or surgical approaches carried out in treatment modalities for bisphosphonate related necrosis of jaws (BRONJ), the most common side effect of bisphosphonates used to inhibit bone resorption. The purpose of this study was to investigate the effects of LLLT on cell proliferation and alkaline phosphatase (ALP) activity of human osteoblast-like cells (Saos-2) treated with different doses of zoledronate, the most potent bisphosphonate. Saos-2 cells were treated with different concentrations of zoledronate and were irradiated with diode laser (wavelength 808 nm, 10 s, 0.25 or 0.50 W). Cell numbers and ALP activity of the cells were determined. LLLT mildly increased the proliferation rate or ALP activity, while zoledronate reduced both. When applied together, LLLT lessened the detrimental effects of zoledronate and improved cell function and/or proliferation. Based on the results of this study, it was concluded that LLLT has biostimulative effects on Saos-2 cells, even after treatment with zoledronate. LLLT may serve as a useful supportive method for BRONJ treatment through enhancement of healing by osteoblasts.

  10. Nickel phosphate molecular sieve as electrochemical capacitors material

    NASA Astrophysics Data System (ADS)

    Yang, Jing-He; Tan, Juan; Ma, Ding

    2014-08-01

    The nickel phosphate molecular sieve VSB-5 as an electrode material for supercapacitors is investigated by cyclic voltammetry (CV) and chronopotentiometry in alkaline media. The VSB-5 shows high specific capacitance and excellent cycling stability. The specific capacitance of VSB-5 is 2740 F g-1 at a current density of 3 A g-1 and there is no significant reduction in Coulombic efficiency after 3000 cycles at 30 A g-1. In comparison with mesoporous nickel phosphate NiPO, porous nickel hydroxide and mesoporous nickel oxide, this remarkable electrochemical performance of VSB-5 will make nickel phosphate material a promising new electrode material for high performance supercapacitors.

  11. Inhibitory activity of phosphates on molds isolated from foods and food processing plants.

    PubMed

    Suárez, V B; Frisón, L; de Basílico, M Z; Rivera, M; Reinheimer, J A

    2005-11-01

    Six commercial phosphates were evaluated for inhibition of the growth of 17 molds isolated from food sources. The assays were performed at neutral and natural (without pH adjustment) pH values, and the molds were streaked on plate count agar with added phosphates. Phosphate concentrations of 0.1, 0.3, 0.5, 1.0, and 1.5% (wt/vol) were used, and the MIC was determined. The resistance of molds to phosphates depended on the species. At a neutral pH, Aspergillus ochraceus and Fusarium proliferatum were resistant to all phosphates at all concentrations assayed, and Byssochlamys nivea, Aureobasidium pullulans, and Penicillium glabrum were most sensitive. The most inhibitory phosphates were those with chain lengths greater than 15 phosphate units and the highest sequestering power. At natural pH values (resulting from dissolving the phosphate in the medium), inhibitory activity changed dramatically for phosphates that produced alkaline or acidic pH in the medium. Phosphates with alkaline pH values (sodium tripolyphosphate of high solubility, sodium tripolyphosphate, and sodium neutral pyrophosphate) were much more inhibitory than phosphates at a neutral pH, but sodium acid pyrophosphate (acidic pH) had decreased inhibitory activity. The results indicate that some phosphates could be used in the food industry to inhibit molds linked to food spoilage.

  12. Autoimmune Lymphoproliferative Syndrome (ALPS) in a Boy with Massive Lymphadenopathy.

    PubMed

    Kianifar, Hamid Reza; Khalesi, Maryam; Farid, Reza; Badiee, Zahra; Rastin, Maryam; Ahanchian, Hamid

    2010-09-01

    Autoimmune lymphoproliferative syndrome (ALPS) is an uncommon nonmalignant lymphoproliferative disease which is characterized by chronic, persistent or recurrent lymphadenopathy, splenomegaly, hepatomegaly, immune cytopenia , hypergammaglobinemia and increased risk of lymphoma. We report a 2-year old boy with hepatosplenomegaly as first presentation. Petechial and purpuric rashes with massive cervical lymphadenopathies developed 10 months later.In laboratory tests anemia, thrombocytopenia and hypergammaglobinemia were observed. According to flocytometry increased double negative T cells and by apoptosis assay decrease apoptosis of lymphocytes accompanied clinical manifestations, thus diagnosis of ALPS was established. In conclusion; in all patients with massive lymphadenopathy and hepatosplenomegay; especially with cytopenia; ALPS should be considered.

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

    SciTech Connect

    Sakisaka, Yukihiko; Tsuchiya, Masahiro; Nakamura, Takashi; Tamura, Masato; Shimauchi, Hidetoshi; Nemoto, Eiji

    2015-08-01

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

  14. Holocene fault scarps in the Western Alps

    NASA Astrophysics Data System (ADS)

    Hippolyte, J. C.

    2003-04-01

    In the Tarentaise Valley, Goguel (1969) had described recent fault scarps. The present work shows that they are normal faults indicating a SE-directed trend of extension in agreement with recent microseismicity data (Sue et al., 1999). It is proposed that they reflect the Quaternary normal reactivation of the "Front du Houiller" thrust fault. In the Belledonne external crystalline massif, Bordet (1970) had observed from helicopter three main fault scarps that he interpreted as active SE-dipping reverse faults. Partly owing to the difficulties of access this area was not visited until now. Field observations reveal that these faults dip in fact 61-68° to the NW, and are normal faults. The faults scarps are 1 to 13 meters high. These faults, together with at least 10 newly discovered conjugate SE-dipping normal fault scarps of 0.5 to 18 meters high, form an about 2 km wide fault zone along the "Synclinal Median" (S.M.) fault. They attest for the activity of this 70 km-long NNE-striking main fault running in the middle of the Belledonne Massif. Its activity is confirmed by major faceted spurs at the La Perche, the La Perrière and the Claran passes, and by ruptures cutting moraines. Other fault scarps are discovered in the whole Belledonne massif showing in particular that the Font-de-France fault, a 60 km-long SE-dipping fault, is also active. All the observed active faults are normal. Their offsets of mountains slopes, of screes and of rock glacier morphologies demonstrate their activity during the Holocene. They indicate a present SE-directed extension in agreement with recent GPS data (Calais et al., 2002). This mapping shows that the present extensional deformation of the Alps is not limited to the west by the "Frontal Pennine thrust" (Sue et al., 1999) but affects also the external Alps. Taking into account focal plane mechanisms, extension affects at least 70 % of the Western Alps. Some scarps have been sampled for Beryllium cosmogenic dating. However

  15. Paleotopography of the Miocene European Central Alps

    NASA Astrophysics Data System (ADS)

    Campani, M.; Mulch, A.; Kempf, O.; Schlunegger, F.; Mancktelow, N.

    2011-12-01

    Reconstructing the surface elevation, surface uplift, and relief evolution histories is fundamental to understanding the growth of mountain ranges, to explore their topographic limits, and relate these to geodynamic and Earth surface processes. Recent geologic and geodynamic models for the Central European Alps propose that the bulk of topography was built through the Pliocene, mainly based on the observation of a strong increase in sedimentation and erosion rates during the last 5-6 Ma, suggesting that the Alps never attained elevations as high as today. Here, we aim to quantify the Miocene (20-14 Ma) paleoelevation of the Central Alps through stable isotope paleoaltimetry. The novelty of the approach presented here, which renders it rather insensitive to past climate change, is to analyze stable isotope proxies of identical age, both from high internal parts of the Alpine orogen and from the adjacent foreland basin that was at or near sea level. We first exploit the hydrogen isotopic ratio in phyllosilicates (mica and chlorite) that interacted with meteoric water during activity of the Simplon detachment, a major normal fault that developed during orogen-parallel extension in high elevation regions. We then contrast the resulting meteoric water compositions with those recorded in carbonate-bearing paleosols of the North-Alpine foreland basin to provide an estimate of relative elevation differences. In the North-Alpine foreland basin, we present oxygen and carbon isotopic compositions of pedogenic mudstones and carbonate concretions. These terrestrial paleosols, dated with ca. 100 ka precision, serve as our point of reference for stable isotope paleoaltimetry, since they formed at or near sea level. Here, δ18O and δ13C values vary between +19 to +25% (SMOW) and -7 to +1% respectively and show close correspondence to global climate change during the mid-Miocene climatic optimum. In the Simplon fault zone, detachment-related muscovite (-126%) and chlorite (-135

  16. Escherichia coli alkaline phosphatase. Kinetic studies with the tetrameric enzyme.

    PubMed

    Halford, S E; Schlesinger, M J; Gutfreund, H

    1972-03-01

    1. The stability of the tetrameric form of Escherichia coli alkaline phosphatase was examined by analytical ultracentrifugation. 2. The stopped-flow technique was used to study the hydrolysis of nitrophenyl phosphates by the alkaline phosphatase tetramer at pH7.5 and 8.3. In both cases transient product formation was observed before the steady state was attained. Both transients consisted of the liberation of 1mol of nitrophenol/2mol of enzyme subunits within the dead-time of the apparatus. The steady-state rates were identical with those observed with the dimer under the same conditions. 3. The binding of 2-hydroxy-5-nitrobenzyl phosphonate to the alkaline phosphatase tetramer was studied by the temperature-jump technique. The self-association of two dimers to form the tetramer is linked to a conformation change within the dimer. This accounts for the differences between the transient phases in the reactions of the dimer and the tetramer with substrate. 4. Addition of P(i) to the alkaline phosphatase tetramer caused it to dissociate into dimers. The tetramer is unable to bind this ligand. It is suggested that the tetramer undergoes a compulsory dissociation before the completion of its first turnover with substrate. 5. On the basis of these findings a mechanism is proposed for the involvement of the alkaline phosphatase tetramer in the physiology of E. coli.

  17. 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.

  18. 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.

  19. Comparison of Osteogenesis between Adipose-Derived Mesenchymal Stem Cells and Their Sheets on Poly-ε-Caprolactone/β-Tricalcium Phosphate Composite Scaffolds in Canine Bone Defects

    PubMed Central

    Kim, Yongsun; Lee, Seung Hoon; Kang, Byung-jae; Kim, Wan Hee; Yun, Hui-suk

    2016-01-01

    Multipotent mesenchymal stem cells (MSCs) and MSC sheets have effective potentials of bone regeneration. Composite polymer/ceramic scaffolds such as poly-ε-caprolactone (PCL)/β-tricalcium phosphate (β-TCP) are widely used to repair large bone defects. The present study investigated the in vitro osteogenic potential of canine adipose-derived MSCs (Ad-MSCs) and Ad-MSC sheets. Composite PCL/β-TCP scaffolds seeded with Ad-MSCs or wrapped with osteogenic Ad-MSC sheets (OCS) were also fabricated and their osteogenic potential was assessed following transplantation into critical-sized bone defects in dogs. The alkaline phosphatase (ALP) activity of osteogenic Ad-MSCs (O-MSCs) and OCS was significantly higher than that of undifferentiated Ad-MSCs (U-MSCs). The ALP, runt-related transcription factor 2, osteopontin, and bone morphogenetic protein 7 mRNA levels were upregulated in O-MSCs and OCS as compared to U-MSCs. In a segmental bone defect, the amount of newly formed bone was greater in PCL/β-TCP/OCS and PCL/β-TCP/O-MSCs/OCS than in the other groups. The OCS exhibit strong osteogenic capacity, and OCS combined with a PCL/β-TCP composite scaffold stimulated new bone formation in a critical-sized bone defect. These results suggest that the PCL/β-TCP/OCS composite has potential clinical applications in bone regeneration and can be used as an alternative treatment modality in bone tissue engineering. PMID:27610141

  20. 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

  1. Pyrophosphate Stimulates Differentiation, Matrix Gene Expression and Alkaline Phosphatase Activity in Osteoblasts

    PubMed Central

    Pujari-Palmer, Michael; Pujari-Palmer, Shiuli; Lu, Xi; Lind, Thomas; Melhus, Håkan; Engstrand, Thomas; Karlsson-Ott, Marjam; Engqvist, Hakan

    2016-01-01

    Pyrophosphate is a potent mitogen, capable of stimulating proliferation in multiple cell types, and a critical participant in bone mineralization. Pyrophosphate can also affect the resorption rate and bioactivity of orthopedic ceramics. The present study investigated whether calcium pyrophosphate affected proliferation, differentiation and gene expression in early (MC3T3 pre-osteoblast) and late stage (SAOS-2 osteosarcoma) osteoblasts. Pyrophosphate stimulated peak alkaline phosphatase activity by 50% and 150% at 100μM and 0.1μM in MC3T3, and by 40% in SAOS-2. The expression of differentiation markers collagen 1 (COL1), alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) were increased by an average of 1.5, 2, 2 and 3 fold, by high concentrations of sodium pyrophosphate (100μM) after 7 days of exposure in MC3T3. COX-2 and ANK expression did not differ significantly from controls in either treatment group. Though both high and low concentrations of pyrophosphate stimulate ALP activity, only high concentrations (100μM) stimulated osteogenic gene expression. Pyrophosphate did not affect proliferation in either cell type. The results of this study confirm that chronic exposure to pyrophosphate exerts a physiological effect upon osteoblast differentiation and ALP activity, specifically by stimulating osteoblast differentiation markers and extracellular matrix gene expression. PMID:27701417

  2. Extragalactic photon-ALP conversion at CTA energies

    NASA Astrophysics Data System (ADS)

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

    2017-01-01

    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 (losc) 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 lt) 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 (losc ~ l gg lt). We also make analytical predictions for the transfer functions in the quasi-adiabatic regime (losc ll l, lt). Our results are important in light of the upcoming Cherenkov Telescope Array (CTA), and may also be applied to models with non-zero ALP masses.

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

    SciTech Connect

    Sato, Chieri; Iwasaki, Tsuyoshi; Kitano, Sachie; Tsunemi, Sachi; Sano, Hajime

    2012-06-22

    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 murine 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

  4. 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.

  5. Mechanisms for solubilization of various insoluble phosphates and activation of immobilized phosphates in different soils by an efficient and salinity-tolerant Aspergillus niger strain An2.

    PubMed

    Li, Xiaolong; Luo, Lijin; Yang, Jinshui; Li, Baozhen; Yuan, Hongli

    2015-03-01

    Mechanisms for solubilization of different types of phosphates and activation of immobilized phosphates in different types of soils by an efficient fungal strain An2 were explored and evaluated in this study. An2 was isolated from a Chinese cabbage rhizosphere soil and identified as Aspergillus niger. It could fast release up to 1722, 2066, and 2356 mg L(-1) of soluble phosphorus (P) from 1 % Ca3(PO4)2, Mg3(PO4)2, and AlPO4 (Ca-P, Mg-P, and Al-P) and 215 and 179 mg L(-1) from 0.5 % FePO4 and rock phosphate (Fe-P and RP), respectively. HPLC assay demonstrated that An2 mainly secreted oxalic acid to solubilize Ca-P, Mg-P, Al-P, and Fe-P whereas secreted tartaric acid to solubilize RP. Furthermore, An2 could tolerate salinity up to 4 % NaCl without impairing its phosphate-solubilizing ability. The simulation experiments validated that An2 was able to effectively activate immobilized phosphates in general calcareous, acidic, as well as saline-alkali soils with high total P content. This study shows new insights into the mechanisms for microbial solubilization of different types of phosphates and supports the future application of strain An2 in different types of soils to effectively activate P for plants.

  6. 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.

  7. 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.

  8. Phosphate homeostasis and disorders.

    PubMed

    Manghat, P; Sodi, R; Swaminathan, R

    2014-11-01

    Recent studies of inherited disorders of phosphate metabolism have shed new light on the understanding of phosphate metabolism. Phosphate has important functions in the body and several mechanisms have evolved to regulate phosphate balance including vitamin D, parathyroid hormone and phosphatonins such as fibroblast growth factor-23 (FGF23). Disorders of phosphate homeostasis leading to hypo- and hyperphosphataemia are common and have clinical and biochemical consequences. Notably, recent studies have linked hyperphosphataemia with an increased risk of cardiovascular disease. This review outlines the recent advances in the understanding of phosphate homeostasis and describes the causes, investigation and management of hypo- and hyperphosphataemia.

  9. Pre-treatment serum lactate dehydrogenase and alkaline phosphatase as predictors of metastases in extremity osteosarcoma

    PubMed Central

    Marais, Leonard C.; Bertie, Julia; Rodseth, Reitze; Sartorius, Benn; Ferreira, Nando

    2015-01-01

    Background The prognosis of patients with metastatic osteosarcoma remains poor. However, the chance of survival can be improved by surgical resection of all metastases. In this study we investigate the value of serum alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) in predicting the presence of metastatic disease at time of diagnosis. Methods Sixty-one patients with histologically confirmed conventional osteosarcoma of the extremity were included in the study. Only 19.7% of cases presented without evidence of systemic spread of the disease. Pre-treatment serum ALP and LDH were analysed in patients with and without skeletal or pulmonary metastases. Results Serum LDH and ALP levels were not significantly different in patients with or without pulmonary metastases (p=0.88 and p=0.47, respectively). The serum LDH and ALP levels did however differ significantly in patients with or without skeletal metastases (p<0.001 and p=0.02, respectively). The optimal breakpoint for serum LDH as a marker of skeletal metastases was 849 IU/L (AUC 0.839; Sensitivity=0.88; Specificity=0.73). LDH >454 IU/L equated to 100% sensitivity for detected bone metastases (positive diagnostic likelihood ratio (DLR)=1.32). With a cut-off of 76 IU/L a sensitivity of 100% was reached for serum ALP predicting the presence of skeletal metastases (positive DLR=1.1). In a multivariate analysis both LDH ≥850 IU/L (odds ratio [OR]=9; 95% confidence interval (CI) 1.8–44.3) and ALP ≥280 IU/L (OR=10.3; 95% CI 2.1–50.5) were predictive of skeletal metastases. LDH however lost its significance in a multivariate model which included pre-treatment tumour volume. Conclusion In cases of osteosarcoma with LDH >850 IU/L and/or ALP >280 IU/L it may be prudent to consider more sensitive staging investigations for detection of skeletal metastases. Further research is required to determine the value and the most sensitive cut-off points of serum ALP and LDH in the prediction of skeletal metastases. PMID

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

    PubMed Central

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

    2005-01-01

    A 65 kDa GPI (glycosylphosphatidyl-inositol)-anchored ALP (alkaline phosphatase) was characterized as a functional receptor of the Bacillus thuringiensis subsp. israelensis Cry11Aa toxin in Aedes aegypti midgut cells. Two (a 100 kDa and a 65 kDa) GPI-anchored proteins that bound Cry11Aa toxin were preferentially extracted after treatment of BBMV (brush boder membrane vesicles) from Ae. aegypti midgut epithelia with phospholipase C. The 65 kDa protein was further purified by toxin affinity chromatography. The 65 kDa protein showed ALP activity. The peptide-displaying phages (P1.BBMV and P8.BBMV) that bound to the 65 kDa GPI–ALP (GPI-anchored ALP) and competed with the Cry11Aa toxin to bind to BBMV were isolated by selecting BBMV-binding peptide-phages by biopanning. GPI–ALP was shown to be preferentially distributed in Ae. aegypti in the posterior part of the midgut and in the caeca, by using P1.BBMV binding to fixed midgut tissue sections to determine the location of GPI–ALP. Cry11Aa binds to the same regions of the midgut and competed with P1.BBMV and P8.BBMV to bind to BBMV. The importance of this interaction was demonstrated by the in vivo attenuation of Cry11Aa toxicity in the presence of these phages. Our results shows that GPI–ALP is an important receptor molecule involved in Cry11Aa interaction with midgut cells and toxicity to Ae. aegypti larvae. PMID:16255715

  11. Real-time ratiometric fluorescent assay for alkaline phosphatase activity with stimulus responsive infinite coordination polymer nanoparticles.

    PubMed

    Deng, Jingjing; Yu, Ping; Wang, Yuexiang; Mao, Lanqun

    2015-03-03

    This study demonstrates a novel ratiometric fluorescent method for real-time alkaline phosphatase (ALP) activity assay with stimulus responsive infinite coordination polymer (ICP) nanoparticles as the probe. The ICP nanoparticles used in this study are composed of two components; one is the supramolecular ICP network formed with guanine monophosphate (GMP) as the ligand and Tb(3+) as the central metal ion, and the other is a fluorescent dye, i.e., 7-amino-4-methyl coumarin (coumarin) encapsulated into the ICP network. Upon being excited at 315 nm, the ICP network itself emits green fluorescence at 552 nm. Coumarin dye encapsulated in the ICP network emits weak fluorescence at 450 nm upon excitation at the same wavelength (315 nm), and this fluorescence emission becomes strong when the encapsulated dye is released from the network into the solution phase. Hence, we develop a ratiometric fluorescent assay based on the ALP-induced destruction of the supramolecular ICP network and the release of coumarin. This mechanism can be used for real-time ratiometric fluorescent monitoring of ALP activity by continuously measuring the ratio of fluorescent intensity at the wavelength of 552 nm (F552) to that at 450 nm (F450) (F552/F450) in the time-dependent fluorescent spectra of the coumarin@Tb-GMP suspension containing ALP with different activities. Under the experimental conditions employed here, the F552/F450 value is linear with the ALP activity within a range from 0.025 U/mL to 0.2 U/mL. The detection limit is down to 0.010 U/mL (S/N = 3). Moreover, the assay developed here is employed for ALP inhibitor evaluation. This study offers a simple yet sensitive method for real-time ALP activity assay.

  12. Chloroquine Phosphate Oral

    MedlinePlus

    Chloroquine phosphate is in a class of drugs called antimalarials and amebicides. It is used to prevent and treat ... Chloroquine phosphate comes as a tablet to take by mouth. For prevention of malaria in adults, one dose is ...

  13. [DNA degradation during standard alkaline of thermal denaturation].

    PubMed

    Drozhdeniuk, A P; Sulimova, G E; Vaniushin, B F

    1976-01-01

    Essential degradation 8 DNA (up to 10 per cent) with liberation of acid-soluble fragments takes place on the standard alkaline (0,01 M sodium phosphate, pH 12, 60 degrees, 15 min) or thermal (0.06 M sodium phosphate buffer, pH 6.8, 102 degrees C, 15 min) denaturation. This degradation is more or less selective: fraction of low molecular weight fragments, isolated by hydroxyapatite cromatography and eluted by 0.06 M sodium phosphate buffer, pH 6.8 is rich in adenine and thymine and contains about 2 times less 5-methylcytosine than the total wheat germ DNA. The degree of degradation of DNA on thermal denaturation is higher than on alkaline degradation. Therefore while studying reassociation of various DNA, one and the same standard method of DNA denaturation should be used. Besides, both the level of DNA degradation and the nature of the resulting products (fragments) should be taken into account.

  14. Alkaline battery, separator therefore

    NASA Technical Reports Server (NTRS)

    Schmidt, George F. (Inventor)

    1980-01-01

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

  15. Utility of urinary alkaline phosphatase and γ-glutamyl transpeptidase in diagnosing acute kidney injury in dogs.

    PubMed

    Nivy, Ran; Avital, Yochai; Aroch, Itamar; Segev, Gilad

    2017-02-01

    The diagnostic utility of urinary alkaline phosphatase (uALP) and γ-glutamyl transpeptidase (uGGT) activities in naturally occurring acute kidney injury (AKI) was investigated in a heterogeneous group of dogs. The study included client-owned dogs with AKI (n = 32), chronic kidney disease (CKD, n = 13), lower urinary tract infection (LUTI, n = 15) and healthy controls (n = 24). uGGT and uALP activities were normalised to urinary creatinine (uCr) concentrations (uGGT/uCr and uALP/uCr, respectively). uALP/uCr and uGGT/uCr were positively and significantly correlated (r = 0.619, P <0.001), and differed significantly (P ≤ 0.001) among groups, as well as between AKI and LUTI or CKD groups (P < 0.05), but not between the AKI and control groups. Areas under the receiver operator characteristics (ROC) curve for uALP/uCr and uGGT/uCr as predictors of AKI were 0.75 and 0.65, respectively, with optimal cut-off points showing poor to moderate sensitivity (59% for uALP/uCr and 79% for uGGT/uCr) and specificity (59% for uALP/uCr and 75% for uGGT/uCr). Higher cut-off points, with 90% specificity, showed low sensitivity (41% for both uALP/uCr and uGGT/uCr). In conclusion, uALP/uCr is superior to uGGT/uCr as a marker of AKI, but both uGGT/uCr and uALP/uCr have unsatisfactory discriminatory power for diagnosing naturally occurring AKI in dogs and therefore cannot be recommended as sole screening tests for canine AKI. However, both may serve as ancillary, confirmatory, biomarkers for detecting AKI in dogs if appropriate cut-off points with high specificities are used.

  16. 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

  17. Quantum-Mechanical Study on the Catalytic Mechanism of Alkaline Phosphatases.

    PubMed

    Borosky, Gabriela L

    2017-02-21

    Alkaline phosphatases (APs) catalyze the hydrolysis and transphosphorylation of phosphate monoesters. The catalytic mechanism was examined by quantum-mechanical calculations using an active-site model based on the X-ray crystal structure of the human placental AP. Free energies of activation and of reaction for the catalytic steps were evaluated for a series of aryl and alkyl phosphate esters, and the computational results were compared with experimental values available in the literature. Mechanistic observations previously reported in experimental works were rationalized by the present theoretical study, particularly regarding the difference in the rate-determining step between aryl and alkyl phosphates. The formation rate of the covalent phosphoserine intermediate followed a linear free energy relationship (LFER) with the pKa of the leaving group. This LFER, which could be experimentally determined only for less reactive alkyl phosphates, was verified by the present calculations to apply for the entire set of aryl and alkyl phosphate substrates.

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

    PubMed

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

    2007-11-01

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

  19. Chemical nature of alkaline polyphosphate boundary film at heated rubbing surfaces

    NASA Astrophysics Data System (ADS)

    Wan, Shanhong; Tieu, A. Kiet; Zhu, Qiang; Zhu, Hongtao; Cui, Shaogang; Mitchell, David R. G.; Kong, Charlie; Cowie, Bruce; Denman, John A.; Liu, Rong

    2016-05-01

    Alkaline polyphosphate has been demonstrated to be able to reduce significant wear and friction of sliding interfaces under heavy loads (>1 GPa) and elevated temperature (800 °C and above) conditions, e.g. hot metal manufacturing. The chemical composition and fine structure of polyphosphate lubricating film is not well understood as well as the role of alkaline elements within the reaction film at hot rubbing surface. This work makes use of the coupling surface analytical techniques on the alkaline polyphosphate tribofilm, XANES, TOF-SIMS and FIB/TEM. The data show the composition in gradient distribution and trilaminar structure of tribofilm: a shorter chain phosphate overlying a long chain polyphosphate that adheres onto oxide steel base through a short chain phosphate. The chemical hardness model well explains the anti-abrasive mechanism of alkaline polyphosphate at elevated temperatures and also predicts a depolymerisation and simultaneous cross-linking of the polyphosphate glass. The role of alkaline elements in the lubrication mechanism is especially explained. This work firstly serves as a basis for a detailed study of alkaline polyphosphate tribofilm at temperature over 600 °C.

  20. Chemical nature of alkaline polyphosphate boundary film at heated rubbing surfaces

    PubMed Central

    Wan, Shanhong; Tieu, A. Kiet; Zhu, Qiang; Zhu, Hongtao; Cui, Shaogang; Mitchell, David R. G.; Kong, Charlie; Cowie, Bruce; Denman, John A.; Liu, Rong

    2016-01-01

    Alkaline polyphosphate has been demonstrated to be able to reduce significant wear and friction of sliding interfaces under heavy loads (>1 GPa) and elevated temperature (800 °C and above) conditions, e.g. hot metal manufacturing. The chemical composition and fine structure of polyphosphate lubricating film is not well understood as well as the role of alkaline elements within the reaction film at hot rubbing surface. This work makes use of the coupling surface analytical techniques on the alkaline polyphosphate tribofilm, XANES, TOF-SIMS and FIB/TEM. The data show the composition in gradient distribution and trilaminar structure of tribofilm: a shorter chain phosphate overlying a long chain polyphosphate that adheres onto oxide steel base through a short chain phosphate. The chemical hardness model well explains the anti-abrasive mechanism of alkaline polyphosphate at elevated temperatures and also predicts a depolymerisation and simultaneous cross-linking of the polyphosphate glass. The role of alkaline elements in the lubrication mechanism is especially explained. This work firstly serves as a basis for a detailed study of alkaline polyphosphate tribofilm at temperature over 600 °C. PMID:27180956

  1. Mammalian intestinal alkaline phosphatase acts as highly active exopolyphosphatase.

    PubMed

    Lorenz, B; Schröder, H C

    2001-06-11

    Recent results revealed that inorganic polyphosphates (polyP), being energy-rich linear polymers of orthophosphate residues known from bacteria and yeast, also exist in higher eukaryotes. However, the enzymatic basis of their metabolism especially in mammalian cells is still uncertain. Here we demonstrate for the first time that alkaline phosphatase from calf intestine (CIAP) is able to cleave polyP molecules up to a chain length of about 800. The enzyme acts as an exopolyphosphatase degrading polyP in a processive manner. The pH optimum is in the alkaline range. Divalent cations are not required for catalytic activity but inhibit the degradation of polyP. The rate of hydrolysis of short-chain polyP by CIAP is comparable to that of the standard alkaline phosphatase (AP) substrate p-nitrophenyl phosphate. The specific activity of the enzyme decreases with increasing chain length of the polymer both in the alkaline and in the neutral pH range. The K(m) of the enzyme also decreases with increasing chain length. The mammalian tissue non-specific isoform of AP was not able to hydrolyze polyP under the conditions applied while the placental-type AP and the bacterial (Escherichia coli) AP displayed polyP-degrading activity.

  2. 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

  3. 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

  4. 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.

  5. Alkaline phosphatase level in gingival crevicular fluid during treatment with Quad-Helix.

    PubMed

    Delli Mauri, A; Petrini, M; Vitale, D; Tecco, S; Festa, F; Barbato, E; Spoto, G

    2015-01-01

    The aim of this work is to assess the level of the human alkaline phosphatase enzyme (ALP) during palatal expansion with Quad-Helix (QH) appliance. A total of twenty-two orthodontic patients characterized by contraction of the upper jaw, that needed application of a QH in order to treat their condition, were included in this study. Gingival crevicular fluid (GCF) was collected at four different times: before cementation (T0), after two weeks (T1), after four weeks (T2) and after one year (T3) from application of QH. In each patient maxillary first molars, right (UM-right) and left (UM-left), which were connected with bands to QH, were used for testing; first lower molars were used as Controls (LM-right, LM-left). Data show that ALP level in tension sites was proportional to the average increase of the inter-molar distance; on the contrary, the enzymatic level in compression sites was characterized by an inverse trend. The only exception to this phenomenon was recorded after one year (T3), when the increase of ALP level in both sites of tension and compression was ascribed to a mild inflammation due to bacterial plaque accumulation. The level of ALP in control sites was constant for the whole period of observation. The described ALP fluctuations in accordance with the inter-molar distance increment, shows that the main action of QH on bone remodelling was exerted during the fourth week (T2); for this reason, the monitoring of this enzyme could be used as a marker of effective function of the QH appliance.

  6. Crystal structure of rat intestinal alkaline phosphatase--role of crown domain in mammalian alkaline phosphatases.

    PubMed

    Ghosh, Kaushik; Mazumder Tagore, Debarati; Anumula, Rushith; Lakshmaiah, Basanth; Kumar, P P B S; Singaram, Senthuran; Matan, Thangavelu; Kallipatti, Sanjith; Selvam, Sabariya; Krishnamurthy, Prasad; Ramarao, Manjunath

    2013-11-01

    Intestinal alkaline phosphatases (IAPs) are involved in the cleavage of phosphate prodrugs to liberate the drug for absorption in the intestine. To facilitate in vitro characterization of phosphate prodrugs, we have cloned, expressed, purified and characterized IAPs from rat and cynomolgus monkey (rIAP and cIAP respectively) which are important pre-clinical species for drug metabolism studies. The recombinant rat and monkey enzymes expressed in Sf9 insect cells (IAP-Ic) were found to be glycosylated and active. Expression of rat IAP in Escherichia coli (rIAP-Ec) led to ~200-fold loss of activity that was partially recovered by the addition of external Zn(2+) and Mg(2+) ions. Crystal structures of rIAP-Ec and rIAP-Ic were determined and they provide rationale for the discrepancy in enzyme activities. Rat IAP-Ic retains its activity in presence of both Zn(2+) and Mg(2+) whereas activity of most other alkaline phosphatases (APs) including the cIAP was strongly inhibited by excess Zn(2+). Based on our crystal structure, we hypothesized the residue Q317 in rIAP, present within 7 Å of the Mg(2+) at M3, to be important for this difference in activity. The Q317H rIAP and H317Q cIAP mutants showed reversal in effect of Zn(2+), corroborating the hypothesis. Further analysis of the two structures indicated a close linkage between glycosylation and crown domain stability. A triple mutant of rIAP, where all the three putative N-linked glycosylation sites were mutated showed thermal instability and reduced activity.

  7. Why nature chose phosphates.

    PubMed

    Westheimer, F H

    1987-03-06

    Phosphate esters and anhydrides dominate the living world but are seldom used as intermediates by organic chemists. Phosphoric acid is specially adapted for its role in nucleic acids because it can link two nucleotides and still ionize; the resulting negative charge serves both to stabilize the diesters against hydrolysis and to retain the molecules within a lipid membrane. A similar explanation for stability and retention also holds for phosphates that are intermediary metabolites and for phosphates that serve as energy sources. Phosphates with multiple negative charges can react by way of the monomeric metaphosphate ion PO3- as an intermediate. No other residue appears to fulfill the multiple roles of phosphate in biochemistry. Stable, negatively charged phosphates react under catalysis by enzymes; organic chemists, who can only rarely use enzymatic catalysis for their reactions, need more highly reactive intermediates than phosphates.

  8. Alkaline phytase from lily pollen: Investigation of biochemical properties.

    PubMed

    Jog, Sonali P; Garchow, Barry G; Mehta, Bakul Dhagat; Murthy, Pushpalatha P N

    2005-08-15

    Phytases catalyze the hydrolysis of phytic acid (InsP6, myo-inositol hexakisphosphate), the most abundant inositol phosphate in cells. In cereal grains and legumes, it constitutes 3-5% of the dry weight of seeds. The inability of humans and monogastric animals such as swine and poultry to absorb complexed InsP6 has led to nutritional and environmental problems. The efficacy of supplemental phytases to address these issues is well established; thus, there is a need for phytases with a range of biochemical and biophysical properties for numerous applications. An alkaline phytase that shows unique catalytic properties was isolated from plant tissues. In this paper, we report on the biochemical properties of an alkaline phytase from pollen grains of Lilium longiflorum. The enzyme exhibits narrow substrate specificity, it hydrolyzed InsP6 and para-nitrophenyl phosphate (pNPP). Alkaline phytase followed Michaelis-Menten kinetics with a K(m) of 81 microM and V(max) of 217 nmol Pi/min/mg with InsP6 and a K(m) of 372 microM and V(max) of 1272 nmol Pi/min/mg with pNPP. The pH optimum was 8.0 with InsP6 as the substrate and 7.0 with pNPP. Alkaline phytase was activated by calcium and inactivated by ethylenediaminetetraacetic acid; however, the enzyme retained a low level of activity even in Ca2+-free medium. Fluoride as well as myo-inositol hexasulfate did not have any inhibitory affect, whereas vanadate inhibited the enzyme. The enzyme was activated by sodium chloride and potassium chloride and inactivated by magnesium chloride; the activation by salts followed the Hofmeister series. The temperature optimum for hydrolysis is 55 degrees C; the enzyme was stable at 55 degrees C for about 30 min. The enzyme has unique properties that suggest the potential to be useful as a feed supplement.

  9. Advances in the management and understanding of autoimmune lymphoproliferative syndrome (ALPS).

    PubMed

    Teachey, David T; Seif, Alix E; Grupp, Stephan A

    2010-01-01

    Autoimmune lymphoproliferative syndrome (ALPS) is a disorder of T cell dysregulation caused by defective Fas-mediated apoptosis. Patients with ALPS can develop a myriad of clinical manifestations including lymphadenopathy, hepatosplenomegaly, autoimmunity and increased rates of malignancy. ALPS may be more common that originally thought, and testing for ALPS should be considered in patients with unexplained lymphadenopathy, hepatosplenomegaly, and/or autoimmunity. As the pathophysiology of ALPS is better characterized, a number of targeted therapies are in preclinical development and clinical trials with promising early results. This review describes the clinical and laboratory manifestations found in ALPS patients, as well as the molecular basis for the disease and new advances in treatment.

  10. Insights on the upper mantle beneath the Eastern Alps

    PubMed Central

    Bianchi, Irene; Miller, Meghan S.; Bokelmann, Götz

    2014-01-01

    Analyses of Ps and Sp receiver functions from datasets collected by permanent and temporary seismic stations, image a seismic discontinuity, due to a negative velocity contrast across the entire Eastern Alps. The receiver functions show the presence of the discontinuity within the upper mantle with a resolution of tens of kilometers laterally. It is deeper (100–130 km) below the central portion of the Eastern Alps, and shallower (70–80 km) towards the Pannonian Basin and in the Central Alps. Comparison with previous studies renders it likely that the observed discontinuity coincides with the lithosphere–asthenosphere boundary (LAB) east of 15°E longitude, while it could be associated with a low velocity zone west of 15°E. PMID:25843967

  11. Characterization of optical systems for the ALPS II experiment

    NASA Astrophysics Data System (ADS)

    Spector, Aaron D.; Põld, Jan H.; Bähre, Robin; Lindner, Axel; Willke, Benno

    2016-12-01

    ALPS II is a light shining through a wall style experiment that will use the principle of resonant enhancement to boost the conversion and reconversion probabilities of photons to relativistic WISPs. This will require the use of long baseline low-loss optical cavities. Very high power build up factors in the cavities must be achieved in order to reach the design sensitivity of ALPS II. This necessitates a number of different sophisticated optical and control systems to maintain the resonance and ensure maximal coupling between the laser and the cavity. In this paper we report on the results of the characterization of these optical systems with a 20 m cavity and discuss the results in the context of ALPS II.

  12. Characterization of optical systems for the ALPS II experiment.

    PubMed

    Spector, Aaron D; Põld, Jan H; Bähre, Robin; Lindner, Axel; Willke, Benno

    2016-12-12

    ALPS II is a light shining through a wall style experiment that will use the principle of resonant enhancement to boost the conversion and reconversion probabilities of photons to relativistic WISPs. This will require the use of long baseline low-loss optical cavities. Very high power build up factors in the cavities must be achieved in order to reach the design sensitivity of ALPS II. This necessitates a number of different sophisticated optical and control systems to maintain the resonance and ensure maximal coupling between the laser and the cavity. In this paper we report on the results of the characterization of these optical systems with a 20 m cavity and discuss the results in the context of ALPS II.

  13. Insights on the upper mantle beneath the Eastern Alps.

    PubMed

    Bianchi, Irene; Miller, Meghan S; Bokelmann, Götz

    2014-10-01

    Analyses of Ps and Sp receiver functions from datasets collected by permanent and temporary seismic stations, image a seismic discontinuity, due to a negative velocity contrast across the entire Eastern Alps. The receiver functions show the presence of the discontinuity within the upper mantle with a resolution of tens of kilometers laterally. It is deeper (100-130 km) below the central portion of the Eastern Alps, and shallower (70-80 km) towards the Pannonian Basin and in the Central Alps. Comparison with previous studies renders it likely that the observed discontinuity coincides with the lithosphere-asthenosphere boundary (LAB) east of 15°E longitude, while it could be associated with a low velocity zone west of 15°E.

  14. Analysis of the metabolic turnover of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Validation of novel analytical techniques by using 32P-labelled lipids from erythrocytes.

    PubMed Central

    Hawkins, P T; Michell, R H; Kirk, C J

    1984-01-01

    We have developed methods that yield estimates of the 32P content of each of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate, thus extending the information available from studies of the labelling of these lipids in intact cells or membrane preparations. The analyses are undertaken with the deacylated lipids. Assay of the 5-phosphate of phosphatidylinositol 4,5-bisphosphate is achieved by the use, under conditions of first-order kinetics, of a 5-phosphate-specific phosphomonoesterase present in isolated erythrocyte membranes [Downes, Mussat & Michell (1982) Biochem. J. 203, 169-177]. Assay of the 4-phosphate of phosphatidylinositol 4-phosphate and of the total monoester phosphate content (4-phosphate plus 5-phosphate) of phosphatidylinositol 4,5-bisphosphate employs alkaline phosphatase from bovine intestine. The radioactivity of the 1-phosphate is that remaining as organic phosphate after exhaustive alkaline phosphatase treatment. The methodology has been validated by using lipids from human erythrocytes: these contain no 32P in their 1-phosphate. These methods should be of substantial value in studies of the many cells that show rapid hormonal perturbations of phosphatidylinositol 4,5-bisphosphate metabolism. PMID:6326746

  15. Isolation, transcription, and inactivation of the gene for an atypical alkaline phosphatase of Synechococcus sp. strain PCC 7942.

    PubMed Central

    Ray, J M; Bhaya, D; Block, M A; Grossman, A R

    1991-01-01

    The alkaline phosphatase of Synechococcus sp. strain PCC 7942 is 145 kDa, which is larger than any alkaline phosphatase previously characterized and approximately three times the size of the analogous enzyme in Escherichia coli. The gene for the alkaline phosphatase, phoA, was cloned and sequenced, and the protein that it encodes was found to have little similarity to other phosphatases. Some sequence similarities were observed between the Synechococcus sp. strain PCC 7942 alkaline phosphatase, the alpha subunit of the ATPase from bacteria and chloroplasts, and the UshA sugar hydrolase of E. coli. Also, limited sequence similarity was observed between a region of the phosphatase and a motif implicated in nucleotide binding. Interestingly, although the alkaline phosphatase is transported across the inner cytoplasmic membrane and into the periplasmic space, it does not appear to have a cleavable signal sequence at its amino terminus. The half-life of the mRNA encoding the alkaline phosphatase, measured after inhibition of RNA synthesis, is approximately 5 min. Similar kinetics for the loss of alkaline phosphatase mRNA occur upon the addition of phosphate to phosphate-depleted cultures, suggesting that high levels of this nutrient inhibit transcription from phoA almost immediately. The phoA gene also appears to be the first gene of an operon; the largest detectable transcript that hybridizes to a phoA gene-specific probe is 11 kb, over twice the size needed to encode the mature protein. Other phosphate-regulated mRNAs are also transcribed upstream of the phoA gene. Insertional inactivation of phoA results in the loss of extracellular, phosphate-regulated phosphatase activity but does not alter the capacity of the cell for phosphate uptake. Images PMID:1712356

  16. Serum levels of PSA, ALP, ICTP, and BSP in prostate cancer patients and the significance of ROC curve in the diagnosis of prostate cancer bone metastases.

    PubMed

    Wei, R J; Li, T Y; Yang, X C; Jia, N; Yang, X L; Song, H B

    2016-06-03

    Bone metastasis is a common complication in prostate cancer patients that can cause bone pain and pathological fracture. This study tested serum levels of prostate specific antigen (PSA), alkaline phosphatase (ALP), bone sialoprotein (BSP), collagen type I pyridine crosslinking peptide (ICTP) in prostate cancer patients and the significance of the receiver operator characteristic (ROC) curve in the diagnosis of prostate cancer bone metastases. Eighty-three prostate cancer patients were enrolled including 42 in the bone metastases group and 41 in the non-bone metastases group. Serum levels of BSP, ALP, ICTP, and PSA were highest in the bone metastases group followed by the non-bone metastases group, hyperplasia group, and then the control group (P < 0.05). Based on Gleason score, serum levels were highest in the poorly differentiated group followed by moderately differentiated and well-differentiated groups (P < 0.05). ROC curve analysis revealed that the diagnostic efficiency of the biomarkers in turn was BSP, PSA, ICTP, and ALP. The sensitivity of BSP, ALP, ICTP, and PSA in the diagnosis of prostate cancer bone metastases were 80.95, 57.14, 69.05, 71.43%, respectively, and the specificity of the same markers were 72.80, 64.80, 76.80, and 88.80%, respectively. Combined detection of the four markers improved sensitivity to 97.62% and the negative-predictive value increased to 97.60%. PSA + BSP showed the best efficiency when combining two markers. In conclusion, serum levels of BSP, ALP, ICTP, and PSA increased in patients with bone metastases, and combined detection of all markers could improve the positive-predictive value.

  17. MAPK signaling pathway alters expression of midgut ALP and ABCC genes and causes resistance to Bacillus thuringiensis Cry1Ac toxin in diamondback moth.

    PubMed

    Guo, Zhaojiang; Kang, Shi; Chen, Defeng; Wu, Qingjun; Wang, Shaoli; Xie, Wen; Zhu, Xun; Baxter, Simon W; Zhou, Xuguo; Jurat-Fuentes, Juan Luis; Zhang, Youjun

    2015-04-01

    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.

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

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

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

  19. Association of collagen with calcium phosphate promoted osteogenic responses of osteoblast-like MG63 cells.

    PubMed

    Hong, Yoon Jung; Chun, Jae-Sig; Lee, Woo-Kul

    2011-04-01

    In this investigation, the effects of the association of the collagen (COLL) molecules with the calcium phosphate (CaP) film were examined with respect to both the physicochemical properties of the CaP films and the osteoblast responses, such as the adhesion, proliferation, differentiation, and mineralization. The COLL pre-adsorbed CaP film (CaPA) exhibited significant changes in the surface morphology compared to the COLL incorporated CaP film (CaPC). The adhesions of the osteoblast-like MG63 cells were similar on the CaPC or CaPA films. However, the proliferation of the MG63 cells on CaPC was comparable to CaP but considerably different than CaPA. The differentiation of the MG63 cells was greatly improved on CaPC and CaPA compared to CaP and more pronounced on CaPA. The presence of COLL within or on the CaP films significantly modulated the expression of the phenotypic genes, including osteopontin (OPN), alkaline phosphatase (ALP), and the transforming growth factor-β (TGF-β). The expression patterns of these genes elucidated that COLL that was present within or on the CaP film supported the osteoblast proliferation and differentiation. These positive effects were stronger for CaPA than CaPC. The bone-like nodules formed on all of the specimens. However, the mineralization of CaPC and CaPA was significantly higher than CaP, indicating that the association of CaP with COLL promoted the mineral deposition. Therefore, the association of the COLL molecules with the CaP film induced positive effects on the biomineralization. Overall, the incorporation of COLL efficiently enhanced the osteoblast responses of CaP. This system can be utilized in a drug delivery system using calcium phosphate. Although the incorporation effects were slightly higher for the osteoblast responses of CaPA than CaPC, CaPC can be used when the longer drug release times are desirable.

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

    PubMed

    Salter, Robert S; Fitchen, John

    2006-01-01

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

  1. Phosphate, inositol and polyphosphates.

    PubMed

    Livermore, Thomas M; Azevedo, Cristina; Kolozsvari, Bernadett; Wilson, Miranda S C; Saiardi, Adolfo

    2016-02-01

    Eukaryotic cells have ubiquitously utilized the myo-inositol backbone to generate a diverse array of signalling molecules. This is achieved by arranging phosphate groups around the six-carbon inositol ring. There is virtually no biological process that does not take advantage of the uniquely variable architecture of phosphorylated inositol. In inositol biology, phosphates are able to form three distinct covalent bonds: phosphoester, phosphodiester and phosphoanhydride bonds, with each providing different properties. The phosphoester bond links phosphate groups to the inositol ring, the variable arrangement of which forms the basis of the signalling capacity of the inositol phosphates. Phosphate groups can also form the structural bridge between myo-inositol and diacylglycerol through the phosphodiester bond. The resulting lipid-bound inositol phosphates, or phosphoinositides, further expand the signalling potential of this family of molecules. Finally, inositol is also notable for its ability to host more phosphates than it has carbons. These unusual organic molecules are commonly referred to as the inositol pyrophosphates (PP-IPs), due to the presence of high-energy phosphoanhydride bonds (pyro- or diphospho-). PP-IPs themselves constitute a varied family of molecules with one or more pyrophosphate moiety/ies located around the inositol. Considering the relationship between phosphate and inositol, it is no surprise that members of the inositol phosphate family also regulate cellular phosphate homoeostasis. Notably, the PP-IPs play a fundamental role in controlling the metabolism of the ancient polymeric form of phosphate, inorganic polyphosphate (polyP). Here we explore the intimate links between phosphate, inositol phosphates and polyP, speculating on the evolution of these relationships.

  2. Phosphate removal using sludge from fuller's earth production.

    PubMed

    Moon, Yong Hee; Kim, Jae Gon; Ahn, Joo Sung; Lee, Gyoo Ho; Moon, Hi-Soo

    2007-05-08

    This study assesses the phosphate removal capacity and mechanism of precipitation or adsorption from aqueous solutions in batch experiments by an industrial sludge containing gypsum (CaSO(4).2H(2)O) obtained as a by-product from a fuller's earth process. The potential capacity for phosphate removal was tested using various solution concentrations, pH values, reaction times, and amount of sludge. The maximum phosphate adsorption capacity calculated using the Langmuir equation was 2.0 g kg(-1). The pH for the maximum adsorption by the sludge was neutral to alkaline (pH 7-12). Over 99% of phosphate was removed from a phosphate solution of 30 mg L(-1) using 0.15 g of sludge in a 9-h reaction. Sulfate (SO(4)(2-)) concentration increased with increasing initial phosphate concentration, possibly because of dissolution of gypsum and adsorption of both sulfate and phosphate. At high phosphate concentration (>1000 mg L(-1)), relative constant concentration of Ca(2+) was not consistent with adsorption of the most important phosphate removal mechanism. Results suggest that precipitation of calcium phosphate is principally responsible for phosphate removal under its high concentration. Agglomerated precipitate in the reaction sludge was observed by SEM and identified as brushite (CaHPO(4).2H(2)O) by XRD, FT-IR, and DTA. Based on thermodynamic considerations, it is suggested that the brushite will readily transform to more stable phases, such as hydroxyapatite (Ca(5)(PO(4))(3).OH).

  3. Alkaline phosphatase activity in gingival crevicular fluid during human orthodontic tooth movement.

    PubMed

    Perinetti, Giuseppe; Paolantonio, Michele; D'Attilio, Michele; D'Archivio, Domenico; Tripodi, Domenico; Femminella, Beatrice; Festa, Felice; Spoto, Giuseppe

    2002-11-01

    Bone remodeling that occurs during orthodontic tooth movement is a biologic process involving an acute inflammatory response in periodontal tissues. A sequence characterized by periods of activation, resorption, reversal, and formation has been recently described as occurring in both tension and compression tooth sites during orthodontic tooth movement. We used a longitudinal design to investigate alkaline phosphatase (ALP) activity in gingival crevicular fluid (GCF) to assess whether it can serve as a diagnostic aid in orthodontics. Sixteen patients (mean age, 15.5 years) participated in the study. The maxillary first molars under treatment served as the test teeth (TT) in each patient; in particular, 1 first molar was to be retracted and hence was considered the distalized molar (DM), whereas the contralateral molar (CM) was included in the fixed orthodontic appliance but was not subjected to the distal forces. The DM antagonist first molar (AM), free from any orthodontic appliance, was used as the baseline control. The GCF around the experimental teeth was harvested from mesial and distal tooth sites immediately before appliance activation, 1 hour after, and weekly over the following 4 weeks. The clinical gingival condition was evaluated at the baseline and at the end of the experimental term. ALP activity was determined spectrophotometrically at 30 degrees C, and the results were expressed as total ALP activity (mUnits/sample). GCF ALP activity was significantly elevated in the DMs and the CMs as compared with the AMs at 1, 2, 3, and 4 weeks; conversely, in the AMs, GCF ALP activity remained at baseline levels throughout the experiment. Moreover, the enzyme activity in the DMs was significantly greater than in the CMs. In the DMs, a significantly greater ALP activity was observed in sites of tension compared with sites of compression. This difference was not seen with the CMs, in which the enzyme activity increased to the same extent in tension and compression

  4. Characterization of human periodontal ligament cells cultured on three-dimensional biphasic calcium phosphate scaffolds in the presence and absence of L-ascorbic acid, dexamethasone and β-glycerophosphate in vitro

    PubMed Central

    AN, SHAOFENG; GAO, YAN; LING, JUNQI

    2015-01-01

    The aim of this study was to evaluate the effect of porous biphasic calcium phosphate (BCP) scaffolds on the proliferation and osteoblastic differentiation of human periodontal ligament cells (hPDLCs) in the presence and absence of osteogenic inducer (L-ascorbic acid, dexamethasone and β-glycerophosphate). The cell growth within the scaffolds in the absence of osteogenic inducers was studied by cell counting kit-8 (CCK-8) assay and scanning electron microscopy (SEM). Alkaline phosphatase (ALP) activity and osteoblastic differentiation markers of hPDLCs in BCP scaffolds were examined in the presence and absence of osteogenic inducers. The cell number of hPDLCs in the BCP scaffolds was less than that of hPDLCs cultured in microplates (control). SEM images showed that cells successfully adhered to the BCP scaffolds and spread amongst the pores; they also produced abundant extracellular cell matrix. In the presence and absence of osteogenic inducers, the ALP activity of hPDLCs within BCP scaffolds was suppressed in varying degrees at all time-points. In the absence of osteogenic inducers, hPDLCs in BCP scaffolds express significant higher levels of osteopontin (OPN) mRNA than the control, and there were no significant differences for Runx2 and osteocalcin (OCN) mRNA levels compared with those cultured in microplates. In the presence of osteogenic inducers, Runx2 expression levels were significantly higher than those in control. OPN and OCN mRNA levels were downregulated slightly. Three-dimensional porous BCP scaffolds are able to stimulate the osteoblastic differentiation of hPDLCs in the presence and absence of osteogenic inducer and may be capable of supporting hPDLC-mediated bone formation. PMID:26622495

  5. 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.

  6. Zinc phosphate conversion coatings

    DOEpatents

    Sugama, T.

    1997-02-18

    Zinc phosphate conversion coatings for producing metals which exhibit enhanced corrosion prevention characteristics are prepared by the addition of a transition-metal-compound promoter comprising a manganese, iron, cobalt, nickel, or copper compound and an electrolyte such as polyacrylic acid, polymethacrylic acid, polyitaconic acid and poly-L-glutamic acid to a phosphating solution. These coatings are further improved by the incorporation of Fe ions. Thermal treatment of zinc phosphate coatings to generate {alpha}-phase anhydrous zinc phosphate improves the corrosion prevention qualities of the resulting coated metal. 33 figs.

  7. Zinc phosphate conversion coatings

    DOEpatents

    Sugama, Toshifumi

    1997-01-01

    Zinc phosphate conversion coatings for producing metals which exhibit enhanced corrosion prevention characteristics are prepared by the addition of a transition-metal-compound promoter comprising a manganese, iron, cobalt, nickel, or copper compound and an electrolyte such as polyacrylic acid, polymethacrylic acid, polyitaconic acid and poly-L-glutamic acid to a phosphating solution. These coatings are further improved by the incorporation of Fe ions. Thermal treatment of zinc phosphate coatings to generate .alpha.-phase anhydrous zinc phosphate improves the corrosion prevention qualities of the resulting coated metal.

  8. Constraining ALP-photon coupling using galaxy clusters

    NASA Astrophysics Data System (ADS)

    Schlederer, Martin; Sigl, Günter

    2016-01-01

    We study photon-ALP conversion by resonance effects in the magnetized plasma of galaxy clusters and compare the predicted distortion of the cosmic microwave background spectrum in the direction of such objects to measurements of the thermal Sunyaev-Zeldovich effect. Using galaxy cluster models based on current knowledge, we obtain upper limits on the photon-ALP coupling constant g of lesssim Script O(10-11 GeV-1). The constraints apply to the mass range of 2 · 10-14 eV lesssim mALP lesssim 3 · 10-12 eV in which resonant photon-ALP conversions can occur. These limits are slightly stronger than current limits, and furthermore provide an independent constraint. We find that a next generation PRISM-like experiment would allow limits down to g ≈ Script O (10-14 GeV-1), two orders of magnitude stronger than the currently strongest limits in this mass range.

  9. 13 CFR 120.841 - Qualifications for the ALP.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    .... 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...

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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.

  15. Autoimmune lymphoproliferative syndrome (ALPS): a rare cause of immune cytopenia.

    PubMed

    John, M Joseph; Rajasekhar, Reena; Mathews, Vikram

    2008-02-01

    Autoimmune Lymphoproliferative syndrome (ALPS) is an inherited disorder manifesting with autoimmune cytopenia, lymphadenopathy and splenomegaly. The differential diagnosis includes infections, autoimmune disorders or malignancies. The disease is characterized by accumulation of double negative (CD3+ CD4- CD8-) T cells (DNT) in the peripheral blood. We describe a case and review the literature.

  16. 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)

  17. Present-day uplift of the western Alps

    PubMed Central

    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.

    2016-01-01

    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. PMID:27346228

  18. 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…

  19. 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.

  20. Constraining ALP-photon coupling using galaxy clusters

    SciTech Connect

    Schlederer, Martin; Sigl, Günter E-mail: guenter.sigl@desy.de

    2016-01-01

    We study photon-ALP conversion by resonance effects in the magnetized plasma of galaxy clusters and compare the predicted distortion of the cosmic microwave background spectrum in the direction of such objects to measurements of the thermal Sunyaev-Zeldovich effect. Using galaxy cluster models based on current knowledge, we obtain upper limits on the photon-ALP coupling constant g of ∼< O(10{sup −11} GeV{sup −1}). The constraints apply to the mass range of 2 · 10{sup −14} eV ∼< m{sub ALP} ∼< 3 · 10{sup −12} eV in which resonant photon-ALP conversions can occur. These limits are slightly stronger than current limits, and furthermore provide an independent constraint. We find that a next generation PRISM-like experiment would allow limits down to g ≈ O (10{sup −14} GeV{sup −1}), two orders of magnitude stronger than the currently strongest limits in this mass range.

  1. Constraining ALP-photon coupling using galaxy clusters

    SciTech Connect

    Schlederer, Martin; Sigl, Günter

    2016-01-21

    We study photon-ALP conversion by resonance effects in the magnetized plasma of galaxy clusters and compare the predicted distortion of the cosmic microwave background spectrum in the direction of such objects to measurements of the thermal Sunyaev-Zeldovich effect. Using galaxy cluster models based on current knowledge, we obtain upper limits on the photon-ALP coupling constant g of ≲O(10{sup −11} GeV{sup −1}). The constraints apply to the mass range of 2⋅10{sup −14} eV ≲m{sub ALP}≲3⋅10{sup −12} eV in which resonant photon-ALP conversions can occur. These limits are slightly stronger than current limits, and furthermore provide an independent constraint. We find that a next generation PRISM-like experiment would allow limits down to g≈O(10{sup −14} GeV{sup −1}), two orders of magnitude stronger than the currently strongest limits in this mass range.

  2. The influence of Sr content in calcium phosphate coatings.

    PubMed

    Lindahl, Carl; Pujari-Palmer, Shiuli; Hoess, Andreas; Ott, Marjam; Engqvist, Håkan; Xia, Wei

    2015-08-01

    In this study calcium phosphate coatings with different amounts of strontium (Sr) were prepared using a biomineralization method. The incorporation of Sr changed the composition and morphology of coatings from plate-like to sphere-like morphology. Dissolution testing indicated that the solubility of the coatings increased with increased Sr concentration. Evaluation of extracts (with Sr concentrations ranging from 0 to 2.37 μg/mL) from the HA, 0.06Sr, 0.6Sr, and 1.2Sr coatings during in vitro cell cultures showed that Sr incorporation into coatings significantly enhanced the ALP activity in comparison to cells treated with control and HA eluted media. These findings show that calcium phosphate coatings could promote osteogenic differentiation even in a low amount of strontium.

  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. New, rare or remarkable microfungi in the Italian Alps (Carnic Alps)--part I--ascomycotina.

    PubMed

    Feige, G B; Ale-Agha, N; Jensen, M; Christiaans, B; Kricke, R

    2004-01-01

    During our observations in the SE part of the Carnic Alps in the year 2003 we were able to collect and identify 35 ascomycetes on trees and dead wood. Among these one can find numerous ascomycetes of different orders e.g. Pyrenomycetes, Loculoascomycetes and Discomycetes. Some species like Botryosphaeria ribis GROSENLUCHER & DUGGAR on Ribes alpinum L., Dothiora pyrenophora (FR.) FR. on Sorbus aucuparia L., Gemmamyces piceae (BORTH.) CASAGO. on Picea excelsa (LAM.) LINK, Glomerella montana (SACC.) v. ARX & E. MULLER on Sesleria caerulea (L.) ARD, Hymenoscyphus immutabilis (Fuck.) Dennis on Alnus incana (L.) Moench, Hysterographium fraxini (PERS. Ex. FR.) de Not. on Fraxinus ornus L., Lachnellula willkommii (Hartig) DENNIS [= Trichascyphella willkommii (Hartig) NANNF.] on Larix decidua MILL.,Leptosphaeria lycopodina (Mont.) SACC. on Lycopodium annotinum L., Mollisia adenostylidis REHM. on Adenostyles glabra (MILL.) DC., Pezicula cinnamomea (DC.)SACC. [ana: Cryptosporiopsis quercina PETRAK] on Quercus robur L., Pyrenopeziza petiolaris (A. & S. Ex FR.) NANNF. on Acer pseudoplatanus L., Tapesia rosae (PERS.) FUCKEL on Rosa canina L., are new for this area. All specimen are deposited in the Herbarium ESS Mycotheca Parva, Collection G.B. Feige/N. Ale-Agha.

  5. Accelerated Learning Program (ALP): Grade 3-8 Evaluation, 2001-02.

    ERIC Educational Resources Information Center

    Baenen, Nancy; Yaman, Kimberly; Lindblad, Mark

    The Wake County Public Schools, North Carolina (WCPSS), initiated the Accelerated Learning Program (ALP) in 1999-2000 as the major new intervention to help all students reach grade-level performance in reading and mathematics. The ALP program was funded through local and state funds, and in 2001-220, 7,285 students received services through ALP.…

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

    PubMed

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

    2007-04-01

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

  7. Promiscuous sulfatase activity and thio-effects in a phosphodiesterase of the alkaline phosphatase superfamily.

    PubMed

    Lassila, Jonathan K; Herschlag, Daniel

    2008-12-02

    The nucleotide phosphodiesterase/pyrophosphatase from Xanthomonas axonopodis (NPP) is a structural and evolutionary relative of alkaline phosphatase that preferentially hydrolyzes phosphate diesters. With the goal of understanding how these two enzymes with nearly identical Zn(2+) bimetallo sites achieve high selectivity for hydrolysis of either phosphate monoesters or diesters, we have measured a promiscuous sulfatase activity in NPP. Sulfate esters are nearly isosteric with phosphate esters but carry less charge, offering a probe of electrostatic contributions to selectivity. NPP exhibits sulfatase activity with k(cat)/K(M) value of 2 x 10(-5) M(-1) s(-1), similar to the R166S mutant of alkaline phosphatase. We further report the effects of thio-substitution on phosphate monoester and diester reactions. Reactivities with these noncognate substrates illustrate a reduced dependence of NPP reactivity on the charge of the nonbridging oxygen situated between the Zn(2+) ions relative to that in alkaline phosphatase. This reduced charge dependence can explain about 10(2) of the 10(7)-fold differential catalytic proficiency for the most similar monoester and diester substrates in the two enzymes. The results further suggest that active site contacts to substrate oxygen atoms that do not contact the Zn(2+) ions may play an important role in defining the selectivity of the enzymes.

  8. 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

  9. Regional Ambient Noise Tomography in the Eastern Alps of Europe

    NASA Astrophysics Data System (ADS)

    Behm, Michael; Nakata, Nori; Bokelmann, Götz

    2016-08-01

    We present results from ambient noise tomography applied to temporary seismological stations in the easternmost part of the Alps and their transition to the adjacent tectonic provinces (Vienna Basin, Bohemian Massif, Southern Alps, Dinarides). By turning each station into a virtual source, we recover surface waves in the frequency range between 0.1 and 0.6 Hz, which are sensitive to depths of approximately 2-15 km. The utilization of horizontal components allows for the analysis of both Rayleigh and Love waves with comparable signal-to-noise ratio. Measured group wave dispersion curves between stations are mapped to local cells by means of a simultaneous inverse reconstruction technique. The spatial reconstruction for Love-wave velocities fails in the central part of the investigated area, and we speculate that a heterogeneous noise source distribution is the cause for the failure. Otherwise, the obtained group velocity maps correlate well with surface geology. Inversion of Rayleigh-wave velocities for shear-wave velocities along a vertical N-S section stretching from the Bohemian Massif through the Central Alps to the Southern Alps and Dinarides reveals a mid-crustal low-velocity anomaly at the contact between the Bohemian Massif and the Alps, which shows a spatial correlation with the P-wave velocity structure and the low-frequency component of the magnetic anomaly map. Our study is validated by the analysis of resolution and accuracy, and we further compare the result to shear-wave velocity models estimated from other active and passive experiments in the area.

  10. 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.

  11. Slab Detachment Under the Eastern Alps Seen By Seismic Anisotropy

    NASA Astrophysics Data System (ADS)

    Qorbani, E.; Bianchi, I.; Bokelmann, G.

    2014-12-01

    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.

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

    NASA Astrophysics Data System (ADS)

    Qorbani, Ehsan; Bianchi, Irene; Bokelmann, Götz

    2015-04-01

    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.

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

    PubMed

    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.

  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. Fabrication and evaluation of a new composite composed of tricalcium phosphate, gelatin, and Chinese medicine as a bone substitute.

    PubMed

    Yao, Chun-Hsu; Tsai, Hung-Ming; Chen, Yueh-Sheng; Liu, Bai-Shuan

    2005-11-01

    This study investigates the biological effects of traditional Chinese medicines on the activities of bone cells using rat bone cells. Then, a mixture of a GGT composite, that is, a novel biodegradable composite containing genipin crosslinked gelatin and tricalcium phosphate, and traditional Chinese medicine (TCM) was prepared as a GGT-TCM composite. A cultured neonatal rat calvarias organ was used to measure the potential of GGT-TCM composite for use in promoting the regeneration of defective bone tissue. The mitochondria activity of the bone cells following exposure to various concentrations of crude extracts of five herbal Chinese medicines was measured by colorimetric assay. Biochemical markers, such as alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) titers were analyzed to evaluate the activities of bone cells. Finally, we examined the organ culture units, which were maintained in cultured medium for 5 weeks. Morphology of tissue was observed, and the quantitative evaluation of the regenerated bone was determined. In a bone cells culture experiment, adding Cuscuta chinensis Lam. (TCM-5) to the bone cells culture clearly promoted the proliferation and differentiation of the osteoblasts from their precursor cells; but the reduced amount of TRAP indicated that the medicine significantly inhibited the osteoclasts activities. Opposite bone cell responses were observed when Loranthus parasiticus Merr. (TCM-3) and Achyranthes bidentata Bl. (TCM-4) were added to the bone cells culture. Eucommia ulmoides Oliv. (TCM-1) and Dipsacus asper Wall. (TCM-2) potentially influence the proliferation and differentiation of the osteoblasts from their precursor cells, but they did not affect the osteoclasts activities. The finding from the organ culture indicated that Chinese medicine effectively increased the rate of tissue regeneration of damaged bones.

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

    PubMed

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

    2016-12-15

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

  17. CADMIUM PHOSPHATE GLASS

    DOEpatents

    Carpenter, H.W.; Johnson, P.D.

    1963-04-01

    A method of preparing a cadmium phosphate glass that comprises providing a mixture of solid inorganic compounds of cadmuim and phosphate having vaporizable components and heating the resulting composition to a temperature of at least 850 un. Concent 85% C is presented. (AEC)

  18. Bioconjugation of alkaline phosphatase to mechanically processed, aqueous suspendible electrospun polymer nanofibers for use in chemiluminescent detection assays.

    PubMed

    Mark, Sonny S; Stolper, Samuel I; Baratti, Carla; Park, Jason Y; Taku, Maria A; Santiago-Avilés, Jorge J; Kricka, Larry J

    2008-06-11

    Aqueous suspendible polymer nanostructures were prepared by simple microtome processing of electrospun nylon 6 nanofibers and were used to immobilize calf intestinal alkaline phosphatase (ALP) by either covalent or noncovalent bioconjugation chemistries. It was found that noncovalent immobilization of ALP to the mechanically cut nanofibers (mean length approximately 4 microm; mean diameter approximately 80 nm) using a multi-stacked, layer-by-layer (LBL) approach with the cationic polymer Sapphire II resulted in the highest enzyme loading (48.1 +/- 0.4 microg . mg(-1) nanofiber) when compared to other covalent immobilization methods based on glutaraldehyde crosslinking. The biofunctionalized nanofibers were also characterized for their chemiluminescent activity with the dioxetane substrate, CSPD. The results indicate that the kinetic parameters, K(m) and V(max), for the catalytic activity of the nanostructure-bound ALP enzyme were influenced by the particular types of immobilization methods employed. In terms of the overall catalytic performance of the various immobilized ALP systems, a single-stacked LBL assembly approach resulted in the highest level of enzymatic activity per unit mass of nanofiber support. To the best of our knowledge, this study represents the first report examining the preparation of mechanically shortened, aqueous dispersed electrospun polymer nanofibers for potential application as enzyme scaffolds in chemiluminescent-based assay systems.

  19. Alkaline phosphatase activity of Escherichia coli starved in sterile lake water microcosms.

    PubMed

    Ozkanca, R; Flint, K P

    1996-03-01

    Escherichia coli grown in high or low phosphate medium was inoculated into a lake water starvation medium. The viable count decreased at 37 degrees C but not at the lower temperatures over 70 d. Alkaline phosphatase was monitored using a colorimetric assay with pNPP as the substrate. Derepression of the enzyme occurred in cultures starved for > 30 d in the lake water and within 5 d in lake water microcosms supplemented with carbon and nitrogen sources where there was rarely an increase in viable count. Chloramphenicol prevented the synthesis of alkaline phosphatase suggesting that, even under starvation conditions, de novo synthesis of the enzyme occurs.

  20. Stereochemistry of phospho group transfer catalyzed by a mutant alkaline phosphatase

    SciTech Connect

    Butler-Ransohoff, J.E.; Kendall, D.A.; Freeman, S.; Knowles, J.R.; Kaiser, E.T.

    1988-06-28

    The stereochemical course of the phospho group transfer catalyzed by mutant (S102C) alkaline phosphatase from Escherichia coli was investigated by using /sup 31/P nuclear magnetic resonance spectroscopy. Transphosphorylation from 4-nitrophenyl (R/sub P/)-/sup 17/O, /sup 16/O, /sup 18/O)phosphate to (S)-propane-1,2-diol occurs with overall retention of configuration at phosphorus. This result is consistent with the view that the hydrolysis of substrates by this mutant enzyme proceeds by way of a covalent phosphoenzyme intermediate in the same manner as the wild-type alkaline phosphatase.

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

    PubMed Central

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

    1979-01-01

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

  2. Zinc electrode in alkaline electrolyte

    SciTech Connect

    McBreen, J.

    1995-12-31

    The zinc electrode in alkaline electrolyte is unusual in that supersaturated zincate solutions can form during discharge and spongy or mossy zinc deposits can form on charge at low overvoltages. The effect of additives on regular pasted ZnO electrodes and calcium zincate electrodes is discussed. The paper also reports on in situ x-ray absorption (XAS) results on mossy zinc deposits.

  3. 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.

  4. Polymorphisms in FAS and CASP8 genes may contribute to the development of ALPS phenotype: a study in 25 patients with probable ALPS.

    PubMed

    Tan, Çağman; Özgül, Rıza Köksal; Çağdaş Ayvaz, Deniz; Tezcan, İlhan; Sanal, Özden

    2015-01-01

    Defects in genes that have role in apoptotic pathways result in development of Autoimmune Lymphoproliferative Syndrome (ALPS) and ALPS related disorders. Germline and somatic FAS mutations, FASL and CASP10 mutations constitute other genetic defects in ALPS. Patients who fulfill ALPS diagnostic criteria and do not have any identified known disease causing mutations are classified as ALPS-unknown or ALPS phenotype and comprise about one third of all patients. CASP8, NRAS and KRAS gene mutations were reported for ALPS related diseases. We performed DNA sequence analysis in 25 unrelated patients with probable ALPS for FAS, FASL and CASP8 gene defects. Pathogenic mutations could not be found in the FAS, FASL and CASP8 genes. However, we found that the frequencies of SNPs rs2234978 and rs1045487 of FAS and CASP8 genes were significantly higher in the patients. Our results suggest that CASP8 and FAS gene polymorphisms in particular, may contribute to the susceptibility to development of ALPS phenotype.

  5. 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

  6. Late-orogenic vertical movements within the arc of the SW Alps and Ligurian Alps

    NASA Astrophysics Data System (ADS)

    Bertotti, G.; Mosca, P.

    2009-09-01

    Mainly on the basis of seismic data, we reconstruct magnitude and timing of Miocene to Present vertical movements in the region of the Savigliano basin, an up to 4 km thick succession of Neogene to Quaternary sediments accumulated inside the Alpine arc at the western termination of the Po Plain. We focus, in particular, on the margins of the basin which are the link between the mountain range and the subsiding basin. Seismic data from these regions allow for the definition of upward and downward vertical movements (uplift-exhumation and subsidence) with a spatial and temporal resolution which cannot be attained by other methods. The eastern, southern and western margins of the Savigliano basin display very different geometries of the sedimentary bodies and, therefore, document different vertical movements during Miocene to Present times. The eastern part of the Savigliano basin, and the regions to the E, experienced subsidence followed by exhumation. The southern margin of the basin, in the Cuneo area, was much more stable with Miocene to present formations all pinching-out close to the present day outcropping position of the basement-sedimentary cover contact. In the W, seismic sections record a stage of tilting until the Burdigalian, followed by generalized subsidence which affected also the adjacent regions of the SW Alps. The overall pattern of vertical movements is essentially controlled by the NW-ward migration of a syncline-anticline couple with the synclinal axis presently located in the center of the Savigliano basin. The width of the folds is in the order of 30-60 km (1/2 wavelength). In map view, the trace of the fold axes changes from N-S in the Savigliano basin to E-W in the Alessandria basin.

  7. Numerous phosphates of microtubule-associated protein 2 in living rat brain

    SciTech Connect

    Tsuyama, S.; Terayama, Y.; Matsuyama, S.

    1987-08-05

    Microtubule-associated protein 2 (MAP 2) purified from microwave-irradiated rat head contained about 46 esterified phosphates (mole/mol), which were not bound covalently to lipids and did not assemble with microtubules. After some phosphates were released by calf intestinal alkaline phosphatase, the phosphate content of MAP-2 decreased to 16 mol of phosphate and the protein assembled in vitro. MAP-2 purified after microtubule assembly cycles and also the cytosolic heat-stable fraction without assembly cycles had 10 mol of phosphate, and both assembled with microtubules. The MAP-2 with 46 phosphates and that with 10 had different pI in isoelectric focusing, but the components, MAP-2a and -2b, were always near each other. In high-pressure liquid chromatography, MAP-2 containing 46 mol of phosphate appeared after that 10 mol of phosphate. Phosphoserine, phosphothreonine, and phosphotyrosine were recovered from tryptic digestion of MAP-2 with 46 mol of phosphate. These findings suggest that two kinds of MAP-2, one with 46 phosphates and not bound to tubulin and the other with 10-16 phosphates and bound to tubulin, are present in the living rat brain.

  8. PHOSPHATE MANAGEMENT: FY2010 RESULTS OF PHOSPHATE PRECIPITATION TESTS

    SciTech Connect

    Hay, M.; King, W.

    2011-04-04

    The Phosphate Management program seeks to develop treatment options for caustic phosphate solutions resulting from the caustic leaching of the bismuth phosphate sludge. The SRNL subtask investigated the precipitation of phosphate salts from caustic solutions through addition of fluoride and by crystallization. The scoping tests examined the: precipitation of phosphate by the addition of sodium fluoride to form the sodium fluorophosphate double salt, Na{sub 7}F(PO{sub 4}){sub 2} {center_dot} 19H{sub 2}O, crystallization of phosphate by reducing the temperature of saturated phosphate solutions, and combinations of precipitation and crystallization. A simplified leachate simulant was used in the study produced by dissolving sodium phosphate in 1 M to 3.5 M sodium hydroxide solutions. The results show that all three processes; precipitation with sodium fluoride, crystallization, and combined precipitation/crystallization can be effective for removing large amounts of phosphate from solution. The combined process of precipitation/crystallization showed >90% removal of phosphate at all hydroxide concentrations when cooling a non-saturated phosphate solution from 65 C to 25 C. Based on the measured solubility of sodium phosphate, pH adjustment/caustic addition will also remove large amounts of phosphate from solution (>80%). For all three processes, the phosphate concentration in the caustic solution must be managed to keep the phosphate from becoming too concentrated and thereby potentially forming a solid mass of sodium phosphate after an effective phosphate removal process.

  9. Surfactant-enhanced alkaline flooding with weak alkalis

    SciTech Connect

    French, T.R.; Josephson, C.B.

    1991-02-01

    The objective of Project BE4B in FY90 was to develop cost-effective and efficient chemical flooding formulations using surfactant-enhanced, lower pH (weak) alkaline chemical systems. Chemical systems were studied that mitigate the deleterious effects of divalent ions. The experiments were conducted with carbonate mixtures and carbonate/phosphate mixtures of pH 10.5, where most of the phosphate ions exist as the monohydrogen phosphate species. Orthophosphate did not further reduce the deleterious effect of divalent ions on interfacial tension behavior in carbonate solutions, where the deleterious effect of the divalent ions is already very low. When added to a carbonate mixture, orthophosphate did substantially reduce the adsorption of an atomic surfactant, which was an expected result; however, there was no correlation between the amount of reduction and the divalent ion levels. For acidic oils, a variety of surfactants are available commercially that have potential for use between pH 8.3 and pH 9.5. Several of these surfactants were tested with oil from Wilmington (CA) field and found to be suitable for use in that field. Two low-acid crude oils, with acid numbers of 0.01 and 0.27 mg KOH/g of oil, were studied. It was shown that surfactant-enhanced alkaline flooding does have merit for use with these low-acid crude oils. However, each low-acid oil tested was found to behave differently, and it was concluded that the applicability of the method must be experimentally determined for any given low-acid crude oil. 19 refs., 10 figs. 4 tabs.

  10. Low level laser therapy modulates viability, alkaline phosphatase and matrix metalloproteinase-2 activities of osteoblasts.

    PubMed

    Oliveira, Flávia Amadeu de; Matos, Adriana Arruda; Matsuda, Sandra Satiko; Buzalaf, Marília Afonso Rabelo; Bagnato, Vanderley Salvador; Machado, Maria Aparecida de Andrade Moreira; Damante, Carla Andreotti; Oliveira, Rodrigo Cardoso de; Peres-Buzalaf, Camila

    2017-04-01

    Low level laser therapy (LLLT) has been shown to stimulate bone cell metabolism but their impact on the matrix metalloproteinase (MMP) expression and activity is little explored. This study evaluated the influence of LLLT at two different wavelengths, red and infrared, on MC3T3-E1 preosteoblast viability, alkaline phosphatase (ALP) and MMP-2 and -9 activities. To accomplish this, MC3T3-E1 cells were irradiated with a punctual application of either red (660nm; InGaAIP active medium) or infrared (780nm; GaAlAs active medium) lasers both at a potency of 20mW, energy dose of 0.08 or 0.16J, and energy density of 1.9J/cm(2) or 3.8J/cm(2), respectively. The control group received no irradiation. Cellular viability, ALP and MMP-2 and -9 activities were assessed by MTT assay, enzymatic activity and zymography, respectively, at 24, 48 and 72h. The treatment of cells with both red and infrared lasers significantly increased the cellular viability compared to the non-irradiated control group at 24 and 48h. The ALP activity was also up modulated in infrared groups at 24 and 72h, depending on the energy densities. In addition, the irradiation with red laser at the energy density of 1.9J/cm(2) promoted an enhancement of MMP-2 activity at 48 and 72h. However, no differences were observed for the MMP-9 activity. In conclusion, when used at these specific parameters, LLL modulates both preosteoblast viability and differentiation highlighted by the increased ALP and MMP-2 activities induced by irradiation.

  11. Association between alendronate, serum alkaline phosphatase level, and heterotopic ossification in individuals with spinal cord injury

    PubMed Central

    Ploumis, Avraam; Donovan, Jayne M.; Olurinde, Mobolaji O.; Clark, Dana M.; Wu, Jason C.; Sohn, Douglas J.; O'Connor, Kevin C.

    2015-01-01

    Context/objective Only sparse evidence exists regarding the effectiveness of oral alendronate (ALN) in the prevention of heterotopic ossification (HO) in patients with spinal cord injury (SCI). The objective of this study is to investigate the protective effect of oral ALN intake on the appearance of HO in patients with SCI. Study design Retrospective database review. Setting A Spinal Cord Unit at a Rehabilitation Hospital. Participants Two hundred and ninety-nine patients with SCI during acute inpatient rehabilitation. Interventions Administration of oral ALN. Outcome measures The incidence of HO during rehabilitation was compared between patients with SCI receiving oral ALN (n = 125) and patients with SCI not receiving oral ALN (n = 174). The association between HO and/or ALN intake with HO risk factors and biochemical markers of bone metabolism were also explored. Results HO developed in 19 male patients (6.35%), however there was no significant difference in the incidence of HO in patients receiving oral ALN or not. The mean odds ratio of not developing versus developing HO given ALN exposure was 0.8. Significant correlation was found between abnormal serum alkaline phosphatase (ALP) levels and HO appearance (P < 0.001) as well as normal serum ALP and ALN intake (P < 0.05). Conclusion Even though there was no direct prevention of HO in patients with SCI by oral ALN intake, abnormal serum ALP was found more frequently in patients with HO development and without oral ALN intake. This evidence could suggest that ALN may play a role in preventing HO, especially in patients with acute SCI with increasing levels of serum ALP. PMID:24820653

  12. Prognostic impact of alkaline phosphatase measured at time of presentation in patients undergoing primary percutaneous coronary intervention for ST-segment elevation myocardial infarction

    PubMed Central

    Kim, Tae-Hoon; Moon, Jeonggeun; Park, Hyun Woo; Jang, Ho-Jun; Park, Sang-Don; Kwon, Sung Woo; Suh, Jon

    2017-01-01

    Background Serum alkaline phosphatase (ALP) has been shown to be a prognostic factor in several subgroups of patients due to its promotion of vascular calcification. However, the prognostic impact of serum ALP level in ST-segment elevation myocardial infarction (STEMI) patients with a relatively low calcification burden has not been determined. We aimed to investigate the association of ALP level measured at time of presentation on clinical outcomes in patients with STEMI requiring primary percutaneous coronary intervention (PCI). Methods A total of 1178 patients with STEMI undergoing primary PCI between 2007 and 2014 were retrospectively enrolled from the INTERSTELLAR registry and classified into tertiles by ALP level (<64, 65–82, or >83 IU/L). The primary study outcome was a major adverse cardiac or cerebrovascular event (MACCE), defined as the composite of all-cause death, non-fatal myocardial infarction, non-fatal stroke, and ischemia-driven revascularization. Results Median follow-up duration was 25 months (interquartile range, 10–39 months). The incidence of MACCE significantly increased as ALP level increased, that is, for the <64, 65–82, and >83 IU/L tertiles incidences were 8.7%, 11.7%, and 15.7%, respectively; p for trend = 0.003). After adjustment for potential confounders, the adjusted hazard ratios for MACCE in the middle and highest tertiles were 1.69 (95% CI 1.01–2.81) and 2.46 (95% CI 1.48–4.09), respectively, as compared with the lowest ALP tertile. Conclusions Elevated ALP level at presentation, but within the higher limit of normal, was found to be independently associated with higher risk of MACCE after primary PCI in patients with STEMI. PMID:28182682

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

    PubMed

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

    2002-07-12

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

  14. Levels of transaminases, alkaline phosphatase, and protein in tissues of Clarias gariepienus fingerlings exposed to sublethal concentrations of cadmium chloride.

    PubMed

    Velmurugan, Babu; Selvanayagam, Mariadoss; Cengiz, Elif I; Uysal, Ersin

    2008-12-01

    The freshwater fish, Clarias gariepienus fingerlings, were exposed to sublethal concentrations (1.7 and 3.4 mg/L) of cadmium chloride for 12 days. Aspartate aminotransferase (AAT), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and total protein levels were assayed in the gill, brain, and muscle of the fish at regular intervals of 6 and 12 days. The activities of AAT, ALT, and ALP of the treated fishes increased significantly in all the tissues compared with the control fish. Protein level in all the tissues showed a significant decrease in comparison to unexposed controls throughout the experimental periods. These results revealed that cadmium chloride effects the intermediary metabolism of C. gariepienus fingerlings and that the assayed enzymes can work as good biomarkers of contamination.

  15. Real-time fluorescence assays of alkaline phosphatase and ATP sulfurylase activities based on a novel PPi fluorescent probe.

    PubMed

    Wang, Xiaobo; Zhang, Zhiyang; Ma, Xiaoyan; Wen, Jinghan; Geng, Zhirong; Wang, Zhilin

    2015-05-01

    An anthracene-armed tetraaza macrocyclic fluorescent probe 3-(9-anthrylmethyl)-3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene(l) for detecting Zn(2+) in aqueous medium was synthesized. L-Zn(2+) complex, showed selectivity toward pyrophosphate ion (PPi) by quenching the fluorescence in aqueous HEPES buffer (pH 7.4). Furthermore, L-Zn(2+) was also used to set up a real-time fluorescence assay for monitoring enzyme activities of alkaline phosphatase (ALP) and adenosine triphosphate sulfurylase (ATPS). In the presence of ALP inhibitor Na3VO4 and ATPS inhibitor chlorate, two enzymes activities decreased obviously, respectively.

  16. Aloe-emodin induces in vitro G2/M arrest and alkaline phosphatase activation in human oral cancer KB cells.

    PubMed

    Xiao, Bingxiu; Guo, Junming; Liu, Donghai; Zhang, Shun

    2007-10-01

    Aloe-emodin is a natural anthraquinone compound from the root and rhizome of Rheum palmatum. In this study, KB cells were treated with 2.5, 5, 10, 20, and 40 microM aloe-emodin for 1 to 5 days. The results showed that aloe-emodin inhibited cancer cells in a dose-dependent manner. Treatment with aloe-emodin at 10 to 40 microM resulted in cell cycle arrest at G2/M phase. The alkaline phosphatase (ALP) activity in KB cells increased upon treatment with aloe-emodin when compared to controls. This is one of the first studies to focus on the expression of ALP in human oral carcinomas cells treated with aloe-emodin. These results indicate that aloe-emodin has anti-cancer effect on oral cancer, which may lead to its use in chemotherapy and chemopreventment of oral cancer.

  17. Skeletal alkaline phosphatase as a serum marker of bone metastases in the follow-up of patients with breast cancer.

    PubMed

    Reale, M G; Santini, D; Marchei, G G; Manna, A; Del Nero, A; Bianco, V; Marchei, P; Frati, L

    1995-01-01

    Immunoradiometric determination of the bone isoenzyme of alkaline phosphatase with a method provided by Hybritech Inc., San Diego CA (USA) was carried out in 145 female patients, 97 of whom with radically operated breast cancer and 48 with benign mammary cysts, in order to evaluate the correlation of serum levels with the metabolic process of bone rearrangement in patients with bone metastases. This study shows that skeletal ALP, having high specificity (86.48%) and sensitivity (78.6%) for early progression (the average anticipation time compared to scintigraphic detection was 101 days) could represent a valid marker for bone metastases in association with mucinous markers in the follow-up of patients operated for breast cancer. In addition, dynamic serum determination of skeletal ALP could be a valid help in monitoring the efficacy of therapy in patients with bone progression.

  18. 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.

  19. 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.

  20. 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

  1. Autoimmune lymphoproliferative syndrome (ALPS). Case report and family history.

    PubMed

    Ries, F; Ferster, A; Rieux-Laucat, F; Biwer, A; Dicato, M

    2010-01-01

    Autoimmune lymphoproliferative syndrome (ALPS) is a rare disease caused by defective lymphocyte apoptosis and is characterized by non-malignant lymphoproliferation, hepatosplenomegaly, autoimmune manifestations and increased risk of both Hodgkin's and non-Hodgkin's lymphoma. Most forms of the disease are due to germ line mutations of the FAS gene and manifest during the first years of life with fluctuating lymphadenopathies, hemolysis, immune thrombocytopenia. During the second decade of life disease manifestations improve spontaneously but autoimmune problems still occur and there is an increased risk of lymphoproliferative malignancy. We describe a typical case of ALPS in a now 44 year old man, followed since the age of 2 for disease manifestations that were unclear at the beginning.

  2. AlpArray-Italy: Site description and noise characterization

    NASA Astrophysics Data System (ADS)

    Govoni, Aladino; D'Alema, Ezio; Capello, Marco; Cavaliere, Stefania Danesi, Adriano; Margheriti, Lucia; Massa, Marco; Mazza, Salvatore; Mazzarini, Francesco; Monna, Stephen; Moretti, Milena; Nardi, Anna; Piccinini, Davide; Piromallo, Claudia; Pondrelli, Silvia; Salimbeni, Simone; Serpelloni, Enrico; Solarino, Stefano; Vallocchia, Massimiliano; Lovati, Sara; Santulin, Marco

    2016-04-01

    Within the framework of the European joint research initiative AlpArray (http://www.alparray.ethz.ch/), we deployed overall 20 seismic broad-band stations in Northern Italy and on two islands in the Tyrrhenian Sea (Capraia and Montecristo) during Fall-Winter 2015. All the stations, connected in real-time, were installed at sites selected according to the AlpArray Seismic Network plan: 16 temporary stations running for two to three years and 4 new permanent stations in sites already occupied by accelerometers of the INGV national network. Most temporary stations are equipped with REF TEK 130S digitizers and Nanometrics Trillium Compact 120s sensors (a couple have Nanometrics Trillium 120P and one Streckeisen STS2). For each site we describe the settings and discuss the noise levels, the site effects and the preliminary sensitivity analysis.

  3. AlpArray Austria & Slovakia: Site description and noise characterization

    NASA Astrophysics Data System (ADS)

    Fuchs, Florian; Kolinsky, Petr; Gröschl, Gidera; Bokelmann, Götz; AlpArray Working Group

    2016-04-01

    Within the framework of the European joint research initiative AlpArray we deployed 30 seismic broadband stations in Eastern Austria and Western Slovakia. Stations were installed between Spring and Winter 2015, with an expected deployment duration of two to three years. Our installations comprising 60s Reftek sensors and Reftek digitizers as well as 3G telemetry are typically located inside shelters like abandoned or rarely used huts and small houses. In this poster, we describe the station setting and surroundings for each of the 30 stations in detail and discuss noise levels and site effects. The description documents technical information for any future studies involving data from temporary AlpArray stations.

  4. Phosphates in some missouri refractory clays

    USGS Publications Warehouse

    Hall, R.B.; Foord, E.E.; Keller, D.J.; Keller, W.D.

    1997-01-01

    . 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.

  5. Bacillus cereus Phosphopentomutase Is an Alkaline Phosphatase Family Member That Exhibits an Altered Entry Point into the Catalytic Cycle*

    PubMed Central

    Panosian, Timothy D.; Nannemann, David P.; Watkins, Guy R.; Phelan, Vanessa V.; McDonald, W. Hayes; Wadzinski, Brian E.; Bachmann, Brian O.; Iverson, Tina M.

    2011-01-01

    Bacterial phosphopentomutases (PPMs) are alkaline phosphatase superfamily members that interconvert α-d-ribose 5-phosphate (ribose 5-phosphate) and α-d-ribose 1-phosphate (ribose 1-phosphate). We investigated the reaction mechanism of Bacillus cereus PPM using a combination of structural and biochemical studies. Four high resolution crystal structures of B. cereus PPM revealed the active site architecture, identified binding sites for the substrate ribose 5-phosphate and the activator α-d-glucose 1,6-bisphosphate (glucose 1,6-bisphosphate), and demonstrated that glucose 1,6-bisphosphate increased phosphorylation of the active site residue Thr-85. The phosphorylation of Thr-85 was confirmed by Western and mass spectroscopic analyses. Biochemical assays identified Mn2+-dependent enzyme turnover and demonstrated that glucose 1,6-bisphosphate treatment increases enzyme activity. These results suggest that protein phosphorylation activates the enzyme, which supports an intermolecular transferase mechanism. We confirmed intermolecular phosphoryl transfer using an isotope relay assay in which PPM reactions containing mixtures of ribose 5-[18O3]phosphate and [U-13C5]ribose 5-phosphate were analyzed by mass spectrometry. This intermolecular phosphoryl transfer is seemingly counter to what is anticipated from phosphomutases employing a general alkaline phosphatase reaction mechanism, which are reported to catalyze intramolecular phosphoryl transfer. However, the two mechanisms may be reconciled if substrate encounters the enzyme at a different point in the catalytic cycle. PMID:21193409

  6. Bacillus cereus Phosphopentomutase Is an Alkaline Phosphatase Family Member That Exhibits an Altered Entry Point into the Catalytic Cycle

    SciTech Connect

    Panosian, Timothy D.; Nannemann, David P.; Watkins, Guy R.; Phelan, Vanessa V.; McDonald, W. Hayes; Wadzinski, Brian E.; Bachmann, Brian O.; Iverson, Tina M.

    2011-09-15

    Bacterial phosphopentomutases (PPMs) are alkaline phosphatase superfamily members that interconvert {alpha}-D-ribose 5-phosphate (ribose 5-phosphate) and {alpha}-D-ribose 1-phosphate (ribose 1-phosphate). We investigated the reaction mechanism of Bacillus cereus PPM using a combination of structural and biochemical studies. Four high resolution crystal structures of B. cereus PPM revealed the active site architecture, identified binding sites for the substrate ribose 5-phosphate and the activator {alpha}-D-glucose 1,6-bisphosphate (glucose 1,6-bisphosphate), and demonstrated that glucose 1,6-bisphosphate increased phosphorylation of the active site residue Thr-85. The phosphorylation of Thr-85 was confirmed by Western and mass spectroscopic analyses. Biochemical assays identified Mn{sup 2+}-dependent enzyme turnover and demonstrated that glucose 1,6-bisphosphate treatment increases enzyme activity. These results suggest that protein phosphorylation activates the enzyme, which supports an intermolecular transferase mechanism. We confirmed intermolecular phosphoryl transfer using an isotope relay assay in which PPM reactions containing mixtures of ribose 5-[{sup 18}O{sub 3}]phosphate and [U-{sup 13}C{sub 5}]ribose 5-phosphate were analyzed by mass spectrometry. This intermolecular phosphoryl transfer is seemingly counter to what is anticipated from phosphomutases employing a general alkaline phosphatase reaction mechanism, which are reported to catalyze intramolecular phosphoryl transfer. However, the two mechanisms may be reconciled if substrate encounters the enzyme at a different point in the catalytic cycle.

  7. 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.

  8. Lichenized and lichenicolous fungi from the Albanian Alps (Kosovo, Montenegro)

    PubMed Central

    Strasser, Eva A.; Hafellner, Josef; Stešević, Danijela; Geci, Fehmi; Mayrhofer, Helmut

    2016-01-01

    396 taxa (381 species) of lichenized and 45 species of lichenicolous fungi from the upper montane, subalpine and alpine belts of the Albanian Alps (= Prokletije Mountain Range, Bjeshkët e Nemuna) are presented. 92 lichenized and 26 lichenicolous fungi are new to Montenegro, 165 lichenized and 24 lichenicolous fungi are new to Kosovo, and 25 lichenized fungi (23 species) are new for the Balkan Peninsula. PMID:26869727

  9. Autoimmune lymphoproliferative syndrome (ALPS): a case with congenital onset.

    PubMed

    Kahwash, Samir B; Fung, Bonita; Savelli, Stephanie; Bleesing, Jack J; Qualman, Stephen J

    2007-01-01

    We describe a case of autoimmune lymphoproliferative syndrome (ALPS), which is very unusual with regard to a clinical onset soon after birth, and a clinical picture dominated by splenomegaly, jaundice, and consumptive peripheral blood cytopenias, with minimal lymphadenopathy. Our documented close follow up demonstrated initial involvement of the spleen, followed by involvement of the bone marrow and the peripheral blood. The patient underwent bone marrow transplant and is alive and well 20 months after diagnosis.

  10. 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

  11. The ALPS Project: Open Source Software for Quantum Lattice Models

    NASA Astrophysics Data System (ADS)

    Trebst, Simon

    2004-03-01

    Algorithms for the simulation of strongly correlated quantum lattice models have matured and there is increasing demand for reliable simulation results both from theoreticians to test ideas and from experimental researchers as means of data analysis. Unlike in other fields there have been no "community codes" available, with the computational experts writing individual codes, adjusting them for specific needs of new projects and thereby investing weeks to months in software development for each project. We will present experiences with the ALPS collaboration, an open source effort aiming at simplifying the development of simulation codes for strongly correlated classical and quantum lattice models. It provides powerful but generic libraries and open-source application programs (such as classical and quantum Monte Carlo, exact diagonalization, DMRG, and others), intended also for non-experts. We will especially address three topics that are of relevance also to other similar efforts: license issues have been extensively discussed, especially concerning the scientific return of making source codes available to the community. The ALPS license is a compromise ensuring scientific return by requesting citations to the original authors of the codes while making sources openly available for future developments. The coordination of an international collaboration with researchers contributing from Austria, France, Germany, Japan and Switzerland by intense developer workshops on a semi-annual basis and annual user workshops is discussed. The situation for funding needed for such a joint open source development effort, which is often classified more as an infrastructure project and less as a research project, is also addressed. Work done with the ALPS collaboration initiated by M. Troyer (ETH) and S. Todo (Tokyo). For details and a list of members see http://alps.comp-phys.org/

  12. Purification and characterization of an extracellular alkaline phosphatase from Penicillium chrysogenum.

    PubMed

    Politino, M; Brown, J; Usher, J J

    1996-01-01

    An extracellular alkaline phosphatase from Penicillium chrysogenum was purified to homogeneity using DEAE ion-exchange chromatography and size exclusion chromatography. SDS-PAGE of the purified enzyme indicated a molecular weight of 58,000. The mobility of the native enzyme on a Superose 12 column suggests that the active form of the enzyme is a monomer. The enzyme catalyzes the hydrolysis of phosphate from a variety of substrates including p-nitrophenyl phosphate, alpha-naphthyl phosphate and the anti-tumor compound etoposide phosphate. The apparent K(m) for the substrate p-nitrophenyl phosphate is 1.3 mM and the enzyme is inhibited by inorganic phosphate. The pH optimum of the enzyme is 9.0 with a broad optimal temperature range between 40 and 50 degrees C. The isoelectric point of the enzyme is approximately 5.5. The enzyme is a glycoprotein; digestion with endoglycosidase H indicates that the protein consists primarily of N-linked carbohydrates. Enzymatic activity is enhanced by the addition of divalent cations such as Mg+2 and Mn+2 and inhibited by addition of a chelator such as EDTA suggesting a metal ion requirement. The enzyme was found to be an inexpensive catalyst for the conversion of etoposide phosphate to etoposide in the manufacture of this anti-tumor compound.

  13. 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.

  14. 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.

  15. A century of leishmaniasis in Alpes-Maritimes, France.

    PubMed

    Marty, P; Izri, A; Ozon, C; Haas, P; Rosenthal, E; Del Giudice, P; Godenir, J; Coulibaly, E; Gari-Toussaint, M; Delaunay, P; Ferrua, B; Haas, H; Pratlong, F; Le Fichoux, Y

    2007-10-01

    A century of publications on leishmaniasis in Alpes-Maritimes, in southern France, is here reviewed. Autochtonous human and canine leishmaniasis were first recognised in this département, which lies by the Mediterranean Sea and near the Italian border, in 1918 and 1925, respectively. The parasite responsible for the leishmaniasis, Leishmania infantum, is transmitted by Phlebotomus perniciosus and P. ariasi. The human leishmaniasis is zoonotic, with domestic dogs acting as the main 'reservoir' hosts. In prospective surveys over the last two decades, a mean of 12% of the domestic dogs checked in Alpes-Maritimes have been found seropositive for L. infantum but only about 50% of the seropositive animals showed any clinical signs of infection at the time of the surveys. During the last 30 years, 178 cases of human visceral leishmaniasis have been recorded in the area. Such cases are sporadic and often opportunistic, occurring predominantly in children (29% of the 178 cases) or HIV-positive subjects (31%). Recently, it has been demonstrated that, in Alpes-Maritimes, approximately 20% of those found seropositive in leishmanin skin tests are asymptomatic carriers, with amastigotes in their peripheral blood.

  16. Biochemical properties and substrate specificities of alkaline and histidine acid phytases.

    PubMed

    Oh, B-C; Choi, W-C; Park, S; Kim, Y-o; Oh, T-K

    2004-01-01

    Phytases are a special class of phosphatase that catalyze the sequential hydrolysis of phytate to less-phosphorylated myo-inositol derivatives and inorganic phosphate. Phytases are added to animal feedstuff to reduce phosphate pollution in the environment, since monogastric animals such as pigs, poultry, and fish are unable to metabolize phytate. Based on biochemical properties and amino acid sequence alignment, phytases can be categorized into two major classes, the histidine acid phytases and the alkaline phytases. The histidine acid phosphatase class shows broad substrate specificity and hydrolyzes metal-free phytate at the acidic pH range and produces myo-inositol monophosphate as the final product. In contrast, the alkaline phytase class exhibits strict substrate specificity for the calcium-phytate complex and produces myo-inositol trisphosphate as the final product. This review describes recent findings that present novel viewpoints concerning the molecular basis of phytase classification.

  17. The secondary alkaline zinc electrode

    NASA Astrophysics Data System (ADS)

    McLarnon, Frank R.; Cairns, Elton J.

    1991-02-01

    The worldwide studies conducted between 1975 and 1990 with the aim of improving cell lifetimes of secondary alkaline zinc electrodes are overviewed. Attention is given the design features and characteristics of various secondary alkaline zinc cells, including four types of zinc/nickel oxide cell designs (vented static-electrolyte, sealed static-electrolyte, vibrating-electrode, and flowing-electrolyte); two types of zinc/air cells (mechanically rechargeable consolidated-electrode and mechanically rechargeable particulate-electrode); zinc/silver oxide battery; zinc/manganese dioxide cell; and zinc/ferric cyanide battery. Particular consideration is given to recent research in the fields of cell thermodynamics, zinc electrodeposition, zinc electrodissolution, zinc corrosion, electrolyte properties, mathematical and phenomenological models, osmotic pumping, nonuniform current distribution, and cell cycle-life perforamnce.

  18. Development of alkaline fuel cells.

    SciTech Connect

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

    2013-09-01

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

  19. Renal threshold phosphate concentration (TmPO4/GFR).

    PubMed Central

    Kruse, K; Kracht, U; Göpfert, G

    1982-01-01

    The ratio of maximum rate of renal tubular reabsorption of phosphate to glomerular filtration rate (TmPO4/GFR) was determined in 546 schoolchildren, aged between 6 and 17.9 years, using the nomogram of Walton and Bijvoet.1 TmPO4/GFR correlated with chronological age in girls and boys and in each remained significantly higher than in adults. TmPO4/GFR in the children correlated neither with fasting serum immunoreactive calcitonin and parathyroid hormone levels nor with the urinary cyclic AMP excretion. The study showed a parallel decrease in TmPO4/GFR, excretion of total hydroxyproline and serum alkaline phosphatase activities after puberty, with a significant relationship of both these indices of bone turnover to TmPO4/GFR values. This indicates that the high renal phosphate threshold of children may be an important factor for bone mineralisation by providing high extracellular inorganic phosphate concentrations during normal growth. PMID:6280622

  20. 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.

  1. 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.

  2. Acute phosphate nephropathy.

    PubMed

    Monfared, Ali; Habibzadeh, Seyed Mahmoud; Mesbah, Seyed Alireza

    2014-05-01

    We present acute phosphate nephropathy in a 28-year-old man, which was developed after a car accident due to rhabdomyolysis. Treatment of acute kidney injury was done with administration of sodium bicarbonate.

  3. Metal-phosphate binders

    DOEpatents

    Howe, Beth Ann [Lewistown, IL; Chaps-Cabrera, Jesus Guadalupe [Coahuila, MX

    2009-05-12

    A metal-phosphate binder is provided. The binder may include an aqueous phosphoric acid solution, a metal-cation donor including a metal other than aluminum, an aluminum-cation donor, and a non-carbohydrate electron donor.

  4. Phosphate control in dialysis

    PubMed Central

    Cupisti, Adamasco; Gallieni, Maurizio; Rizzo, Maria Antonietta; Caria, Stefania; Meola, Mario; Bolasco, Piergiorgio

    2013-01-01

    Prevention and correction of hyperphosphatemia is a major goal of chronic kidney disease–mineral and bone disorder (CKD–MBD) management, achievable through avoidance of a positive phosphate balance. To this aim, optimal dialysis removal, careful use of phosphate binders, and dietary phosphate control are needed to optimize the control of phosphate balance in well-nourished patients on a standard three-times-a-week hemodialysis schedule. Using a mixed diffusive–convective hemodialysis tecniques, and increasing the number and/or the duration of dialysis tecniques are all measures able to enhance phosphorus (P) mass removal through dialysis. However, dialytic removal does not equal the high P intake linked to the high dietary protein requirement of dialysis patients; hence, the use of intestinal P binders is mandatory to reduce P net intestinal absorption. Unfortunately, even a large dose of P binders is able to bind approximately 200–300 mg of P on a daily basis, so it is evident that their efficacy is limited in the case of an uncontrolled dietary P load. Hence, limitation of dietary P intake is needed to reach the goal of neutral phosphate balance in dialysis, coupled to an adequate protein intake. To this aim, patients should be informed and educated to avoid foods that are naturally rich in phosphate and also processed food with P-containing preservatives. In addition, patients should preferentially choose food with a low P-to-protein ratio. For example, patients could choose egg white or protein from a vegetable source. Finally, boiling should be the preferred cooking procedure, because it induces food demineralization, including phosphate loss. The integrated approach outlined in this article should be actively adapted as a therapeutic alliance by clinicians, dieticians, and patients for an effective control of phosphate balance in dialysis patients. PMID:24133374

  5. Phosphate control in dialysis.

    PubMed

    Cupisti, Adamasco; Gallieni, Maurizio; Rizzo, Maria Antonietta; Caria, Stefania; Meola, Mario; Bolasco, Piergiorgio

    2013-10-04

    Prevention and correction of hyperphosphatemia is a major goal of chronic kidney disease-mineral and bone disorder (CKD-MBD) management, achievable through avoidance of a positive phosphate balance. To this aim, optimal dialysis removal, careful use of phosphate binders, and dietary phosphate control are needed to optimize the control of phosphate balance in well-nourished patients on a standard three-times-a-week hemodialysis schedule. Using a mixed diffusive-convective hemodialysis tecniques, and increasing the number and/or the duration of dialysis tecniques are all measures able to enhance phosphorus (P) mass removal through dialysis. However, dialytic removal does not equal the high P intake linked to the high dietary protein requirement of dialysis patients; hence, the use of intestinal P binders is mandatory to reduce P net intestinal absorption. Unfortunately, even a large dose of P binders is able to bind approximately 200-300 mg of P on a daily basis, so it is evident that their efficacy is limited in the case of an uncontrolled dietary P load. Hence, limitation of dietary P intake is needed to reach the goal of neutral phosphate balance in dialysis, coupled to an adequate protein intake. To this aim, patients should be informed and educated to avoid foods that are naturally rich in phosphate and also processed food with P-containing preservatives. In addition, patients should preferentially choose food with a low P-to-protein ratio. For example, patients could choose egg white or protein from a vegetable source. Finally, boiling should be the preferred cooking procedure, because it induces food demineralization, including phosphate loss. The integrated approach outlined in this article should be actively adapted as a therapeutic alliance by clinicians, dieticians, and patients for an effective control of phosphate balance in dialysis patients.

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

    DTIC Science & Technology

    2000-11-13

    106C: 16N: 1P) and alkaline phosphatase activity (APA) were utilized in tandem as nutrient deficiency indicators (NDIs) for phytoplankton . The study...nutrient enrichment incubation re-affirmed the use of APA as a robust indicator of phosphate limitation in phytoplankton . APA data indicate that the system...nutrient deficiency indicators (NDIs) for phytoplankton . The study objective was to evaluate the limiting nutrient status of the pond throughout the

  7. Swiss AlpArray: deployment of the Swiss AlpArray temporary broad-band stations and their noise characterization

    NASA Astrophysics Data System (ADS)

    Molinari, Irene; Kissling, Edi; Clinton, John; Hetényi, György; Šipka, Vesna; Stipćević, Josip; Dasović, Iva; Solarino, Stefano; Wéber, Zoltán; Gráczer, Zoltán; Electronics Lab, SED

    2016-04-01

    One of the main actions of the AlpArray European initiative is the deployment of a dense seismic broad-band network, that complements the existing permanent stations. This will ensure a spatially homogeneous seismic coverage of the greater Alpine area for at least two years, allowing a great number of innovative scientific works to be carried out. Our contribution to the AlpArray Seismic Network consists in the deployment of 24 temporary broad-band stations: three in Switzerland, twelve in Italy, three in Croatia, three in Bosnia and Herzegovina and three in Hungary. This deployment is lead by ETH Zurich and founded by the Swiss-AlpArray Sinergia programme by SNSF, and is the result of a fruitful collaboration between five research institutes. Stations were installed between Autumn and Winter 2015. Our installations are both free field and in-house and consist of 21 STS-2 and 3 Trillium Compact sensors equipped with Taurus digitizers and 3G telemetry sending data in real time to the ETH EIDA node. In this work, we present sites and stations setting and we discuss in details the characteristics in terms of site effects and noise level of each station. In particular we analyse the power spectral density estimates investigating the major source of noise and the background noise related to seasons, time of the day, human activities and type of installation. In addition we will show examples of data usage - i.e. earthquake locations, noise cross correlations, measures of surface wave dispersion curves. We thanks the Swiss AlpArray Field Team: Blanchard A., Erlanger E. D., Jarić D., Herak D., M. Herak, Hermann M., Koelemeijer P. J., Markušić S., Obermann A., Sager K., Šikman S., Singer J., Winterberg S. SED Electronic Lab: Barman S., Graf P., Hansemann R., Haslinger F., Hiemer S., Racine R., Tanner R., Weber F.

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

    NASA Astrophysics Data System (ADS)

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

    2014-09-01

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

  9. Comparative Evaluation of Efficacy of Three Different Herbal Toothpastes on Salivary Alkaline Phosphatase and Salivary Acid Phosphatase - A Randomized Controlled Trial

    PubMed Central

    Dodamani, Arun; Karibasappa, G. N.; Deshmukh, Manjiri; Naik, Rahul

    2016-01-01

    Introduction Very few researches in the past have tried to evaluate the effect of herbal toothpaste on saliva and salivary constituents like alkaline phosphatase and acid phosphatase which play an important role in maintaining oral health. Aim To evaluate and compare the effect of three different herbal toothpastes on Salivary Alkaline Phosphatase (ALP) and salivary Acid Phosphatase (ACP). Material and Methods The present study was a preliminary study conducted among 45 dental students (15 subjects in each group) in the age group of 19-21 years. Subjects in each group were randomly intervened with three different herbal toothpastes respectively (Group A – Patanjali Dant Kanti, Group B - Himalaya Complete Care and Group C – Vicco Vajradanti). Unstimulated saliva sample were collected before and after brushing and salivary ACP and salivary ALP levels were assessed at an interval of one week each for a period of four weeks starting from day one. Compiled data was analyzed using chi square test, paired t-test and ANOVA based on the nature of the obtained data. Results All the three toothpastes showed significant (p<0.001) reduction in ACP and ALP levels at each interval. For patanjali toothpaste, the mean reduction was in the range of 2.55 – 2.62 IU/L for ACP and 2.94 – 2.99 IU/L for ALP. For Himalaya toothpaste, the mean reduction was in the range of 1.39 – 1.47 IU/L for ACP and 1.55 – 1.61 IU/L for ALP. For Vicco toothpaste, the mean reduction was in the range of 2.46 – 2.50 IU/L for ACP and 2.64 – 2.77 IU/L for ALP. Patanjali and Vicco toothpaste were significantly effective in reducing the levels of salivary ACP and ALP more than Himalaya toothpaste (p<0.05). Conclusion Herbal toothpastes, especially Dant Kanti and Vicco Vajradanti, showed significant reduction in levels of ACP and ALP resulting in overall improvement towards the oral health. PMID:27790584

  10. 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.

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

    PubMed

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

    2008-09-01

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

  12. Erosion-driven uplift of the modern Central Alps

    NASA Astrophysics Data System (ADS)

    Champagnac, J.-D.; Schlunegger, F.; Norton, K.; von Blanckenburg, F.; Abbühl, L. M.; Schwab, M.

    2009-04-01

    We present a compilation of four sets of data of modern tectono-geomorphic processes in the Central Alps of Switzerland that appear to suggest that rock uplift is a response to climate-driven denudation in the absence of active convergence. These are (1) basin-averaged Late Holocene denudation rates determined from cosmogenic nuclides and from suspended river loads; these slightly exceed, but spatially mimic the pattern of rock uplift rates as determined by geodetic leveling; (2) the geodetic reference point is also the geomorphic base level with respect to erosion; we further present (3) a compilation of modern plate motion velocities shows that the rotation pole of the Adriatic plate is located within the area, hence the area is not under convergence; finally (4), we illustrate that the Central Alps have acted as a closed system for Holocene sediment redistribution up to the peri-Alpine lakes which have operated as a sink for the erosion products of the inner Alps. While a variety of hypotheses have been put forward to explain the Central Alpine uplift (e.g. lithospheric forcing by convergence or mantle processes; ice melting) we show with a numerical isostatic model that the correlation between erosion and crustal uplift rates reflects a positive feedback between denudation and the associated isostatic response to unloading. Therefore erosion does not passively respond to advection of crustal material as might be the case in actively converging orogens. Other forces need to be considered to drive surface erosion. We suggest that the geomorphic response of the Alpine topography to glacial erosion and the resulting disequilibrium for modern channelized and associated hillslope processes explains much of the pattern of modern denudation and hence rock uplift. Therefore, in a non-convergent orogen such as the Central European Alps, the observed vertical rock uplift is primarily a consequence of passive unloading due to erosion. REFERENCES: Anderson, H. and Jackson

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

    PubMed Central

    Bednarska, Katarzyna; Klink, Magdalena; Sulowska, Zofia

    2006-01-01

    We have proposed the use of the fluorimetric method with 4-methylumbelliferyl phosphate (4-MUP) specific substrate for the alkaline phosphatase determination in the neutrophil adhesion assay. We provide evidence that the endogenous neutrophil alkaline phosphatase (NAP) activity evaluation is reliable to quantify neutrophil adhesion at a wide range of cell numbers (104−106). The results obtained by fluorimetric NAP activity test correlate to the results of adherence evaluated using the MTT reduction assay. The fluorimetric NAP activity test may be applied for resting as well as activated neutrophils without the risk of the activators interferences into the test. The alkaline phosphatase survey with the use of 4-MUP substrate is recommended herein as a sensitive, repeatable, simple, and reliable method of the neutrophil adherence determination in vitro. PMID:17047286

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

    PubMed Central

    Tommassen, J; Lugtenberg, B

    1980-01-01

    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. Images PMID:6995425

  15. Culture of osteogenic cells from human alveolar bone: a useful source of alkaline phosphatase.

    PubMed

    Simão, Ana Maria S; Beloti, Marcio M; Rosa, Adalberto L; de Oliveira, Paulo T; Granjeiro, José Mauro; Pizauro, João M; Ciancaglini, Pietro

    2007-11-01

    The aim of this study was to obtain membrane-bound alkaline phosphatase from osteoblastic-like cells of human alveolar bone. Cells were obtained by enzymatic digestion and maintained in primary culture in osteogenic medium until subconfluence. First passage cells were cultured in the same medium and at 7, 14, and 21 days, total protein content, collagen content, and alkaline phosphatase activity were evaluated. Bone-like nodule formation was evaluated at 21 days. Cells in primary culture at day 14 were washed with Tris-HCl buffer, and used to extract the membrane-bound alkaline phosphatase. Cells expressed osteoblastic phenotype. The apparent optimum pH for PNPP hydrolysis by the enzyme was pH 10.0. This enzyme also hydrolyzes ATP, ADP, fructose-1-phosphate, fructose-6-phosphate, pyrophosphate and beta-glycerophosphate. PNPPase activity was reduced by typical inhibitors of alkaline phosphatase. SDS-PAGE of membrane fraction showed a single band with activity of approximately 120 kDa that could be solubilized by phospholipase C or Polidocanol.

  16. Bioprecipitation of uranium from alkaline waste solutions using recombinant Deinococcus radiodurans.

    PubMed

    Kulkarni, Sayali; Ballal, Anand; Apte, Shree Kumar

    2013-11-15

    Bioremediation of uranium (U) from alkaline waste solutions remains inadequately explored. We engineered the phoK gene (encoding a novel alkaline phosphatase, PhoK) from Sphingomonas sp. for overexpression in the radioresistant bacterium Deinococcus radiodurans. The recombinant strain thus obtained (Deino-PhoK) exhibited remarkably high alkaline phosphatase activity as evidenced by zymographic and enzyme activity assays. Deino-PhoK cells could efficiently precipitate uranium over a wide range of input U concentrations. At low uranyl concentrations (1 mM), the strain precipitated >90% of uranium within 2h while a high loading capacity of around 10.7 g U/g of dry weight of cells was achieved at 10 mM U concentration. Uranium bioprecipitation by Deino-PhoK cells was not affected in the presence of Cs and Sr, commonly present in intermediate and low level liquid radioactive waste, or after exposure to very high doses of ionizing radiation. Transmission electron micrographs revealed the extracellular nature of bioprecipitated U, while X-ray diffraction and fluorescence analysis identified the precipitated uranyl phosphate species as chernikovite. When immobilized into calcium alginate beads, Deino-PhoK cells efficiently removed uranium, which remained trapped in beads, thus accomplishing physical separation of precipitated uranyl phosphate from solutions. The data demonstrate superior ability of Deino-PhoK, over earlier reported strains, in removal of uranium from alkaline solutions and its potential use in bioremediation of nuclear and other waste.

  17. Electrophoretic separation of alkaline and acid phosphatase isoenzymes from the pulp of monkey teeth.

    PubMed

    Franzén, A; Hasselgren, G

    1978-01-01

    Monkey pulps were homogenized in a Triton tris solution. After three centrifugation steps (800, 20000, and 105000 g) the supernatant was applied on acryl amide columns at pH 7.5 in a tris-diethyl barbituric acid buffer. Electrophoresis was performed at a constant current of 2.5 mA per gel column at 18--20 degrees C. Incubations for alkaline phosphatase (E.C. 3.1.3.1) were carried out at pH 8.3 using naphthol-AS-MX-phosphate as substrate and Fast Red Violet LB salt as coupler. Incubations for acid phosphatase (E.C. 3.1.3.2) were undertaken at pH 5.0 using alpha-naphtyl phosphate as substrate and hexazotized pararosanilin as coupling agent. After the incubations for alkaline phosphatase as well as acid phosphatase two bands showing enzyme activity were demonstrated. By means of treatment with heat (56 degrees C) prior to incubation or addition of vanadate or pyrophosphate to the incubation medium it was shown that the main part of the fast moving alkaline phosphatase band was sensitive to these procedures. The alkaline phosphatase of the slow moving band appeared to be resistant to heat or the addition of inhibitors.

  18. 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.

  19. Antibacterial effect of phosphates and polyphosphates with different chain length.

    PubMed

    Lorencová, Eva; Vltavská, Pavlína; Budinský, Pavel; Koutný, Marek

    2012-01-01

    The aim of this study was to monitor the antibacterial effect of seven phosphate salts on selected strains of Gram-negative and Gram-positive bacteria, which could be considered responsible for food-borne diseases (Bacillus cereus, Bacillus subtilis, Enterococcus faecalis, Micrococcus luteus, Staphylococcus aureus, Citrobacter freundii, Escherichia coli, Proteus mirabilis, Salmonella enterica ser. Enteritidis and Pseudomonas aeruginosa). For these purposes, phosphates differing in chain length were used. The tested concentrations were in the range of 0.1-2.0% (wt v(-1)) applied at the model conditions. In the majority of cases the visible inhibitory effect on the growth of observed microorganisms could be seen. Due to the chemical structure of salts and their dissociation both the pH values of cultivation broth and similarly the growth characteristics of bacterial strains were affected. The inhibition of above mentioned bacteria was apparently supported by this dissociation. Phosphates obviously made the development of most Gram-positive bacteria impossible. Especially Micrococcus luteus was extremely sensitive to the presence of these substances. On the other hand, Gram-negative bacteria seemed to be resistant to the phosphate incidence. The exemption clause from the tested salts was represented by a high alkaline trisodium phosphate. It should be pointed out that generally the most significant antibacterial effects were shown by polyphosphates HEXA68 and HEXA70, trisodium phosphate undecahydrate, tetrasodium pyrophosphate and finally trisodium phosphate. By comparing the inhibitory effects of various phosphate salts can be concluded that the antibacterial activity was not determined only by the condensation degree but there was also proved the dependence on pH values.

  20. Effect of NELL1 gene overexpression in iPSC-MSCs seeded on calcium phosphate cement

    PubMed Central

    Liu, Jun; Chen, Wenchuan; Zhao, Zhihe; Xu, Hockin H. K.

    2015-01-01

    Human induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) are a promising source of patient-specific stem cells with great regenerative potential. There has been no report on NEL-like protein 1 (NELL1) gene modification of iPSC-MSCs. The objectives of this study were to genetically modify iPSC-MSCs with NELL1 overexpression for bone tissue engineering, and investigate the osteogenic differentiation of NELL1 gene-modified iPSC-MSCs seeded on Arg-Gly-Asp (RGD)-grafted calcium phosphate cement (CPC) scaffold. Cells were transduced with red fluorescence protein (RFP-iPSC-MSCs) or NELL1 (NELL1-iPSC-MSCs) by a lentiviral vector. Cell proliferation on RGD-grafted CPC scaffold, osteogenic differentiation and bone mineral synthesis were evaluated. RFP-iPSC-MSCs stably expressed high levels of RFP. Both the NELL1 gene and NELL1 protein levels were confirmed higher in NELL1-iPSC-MSCs than in RFP-iPSC-MSCs using RT-PCR and Western blot (p < 0.05). Alkaline phosphatase (ALP) activity was increased by 130% by NELL1 overexpression at 14 d (p < 0.05), indicating that NELL1 promoted iPSC-MSC osteogenic differentiation. When seeded on RGD-grafted CPC, NELL1-iPSC-MSCs attached and expanded similarly well to RFP-iPSC-MSCs. At 14 d, runt-related transcription factor 2 (RUNX2) gene level of NELL1-iPSC-MSCs was 2.0-fold that of RFP-iPSC-MSCs. Osteocalcin (OC) level of NELL1-iPSC-MSCs was 3.1-fold that of RFP-iPSC-MSCs (p < 0.05). Collagen type I alpha 1 (COL1A1) gene level of NELL1-iPSC-MSCs was 1.7-fold that of RFP-iPSC-MSCs at 7 d (p < 0.05). Mineral synthesis was increased by 81% in NELL1-iPSC-MSCs at 21 d. In conclusion, NELL1 overexpression greatly enhanced the osteogenic differentiation and mineral synthesis of iPSC-MSCs on RGD-grafted CPC scaffold for the first time. The novel NELL1-iPSC-MSC seeded RGD-CPC construct is promising to enhance bone engineering. PMID:25220281

  1. Diclofenac salts. III. Alkaline and earth alkaline salts.

    PubMed

    Fini, Adamo; Fazio, Giuseppe; Rosetti, Francesca; Angeles Holgado, M; Iruín, Ana; Alvarez-Fuentes, Josefa

    2005-11-01

    Diclofenac salts containing the alkaline and two earth alkaline cations have been prepared and characterized by scanning electron microscopy (SEM) and EDAX spectroscopy; and by thermal and thermogravimetric analysis (TGA): all of them crystallize as hydrate when precipitated from water. The salts dehydrate at room temperature and more easily on heating, but recovery the hydration, when placed in a humid environment. X-ray diffraction spectra suggest that on dehydration new peaks appear on diffractograms and the lattice of the salts partially looses crystallinity. This phenomenon is readily visible in the case of the calcium and magnesium salts, whose thermograms display a crystallization exotherm, before melting or decomposing at temperatures near or above 200 degrees C; these last salts appear to form solvates, when prepared from methanol. The thermogram of each salt shows a complex endotherm of dehydration about 100 degrees C; the calcium salt displays two endotherms, well separated at about 120 and 160 degrees C, which disappear after prolonged heating. Decomposition exotherms, before or soon after the melting, appear below 300 degrees C. The ammonium salt is thermally unstable and, when heated to start dehydration, dissociates and leaves acidic diclofenac.

  2. Direct nonchromatographic assay for 1-acyl-sn-glycerol-3-phosphate acyltransferase

    SciTech Connect

    Rajasekharan, R.; Ray, T.K.; Cronan, J.E. Jr.

    1988-09-01

    1-Acyl-sn-glycerol-3-phosphate acyltransferase (also called lysophosphatidic acid acyltransferase) which catalyzes the acylation of 1-acyl-sn-glycerol-3-phosphate to phosphatidic acid is generally assayed by the use of a radioactive substrate followed by a time-consuming chromatographic separation of substrate and product. We report a direct and highly sensitive nonchromatographic assay for this enzyme based on the ability of Escherichia coli alkaline phosphatase to dephosphorylate 1-acyl-sn-glycerol-3-phosphate but not phosphatidic acid. This selective hydrolysis coupled with the use of /sup 32/P-labeled 1-acyl-sn-glycerol-3-phosphate as substrate permits measurement of the product, /sup 32/P-labeled phosphatidic acid by solvent extraction or precipitation. We also report a series of enzymatic reactions for the efficient conversion of /sup 32/Pi to /sup 32/P-labeled 1-acyl-sn-glycerol-3-phosphate.

  3. 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

  4. Alkaline fuel cell performance investigation

    NASA Technical Reports Server (NTRS)

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

    1988-01-01

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

  5. 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

  6. MAGNESIUM MONO POTASSIUM PHOSPHATE GROUT FOR P-REACTOR VESSEL IN-SITU DECOMISSIONING

    SciTech Connect

    Langton, C.; Stefanko, D.

    2011-01-05

    The objective of this report is to document laboratory testing of magnesium mono potassium phosphate grouts for P-Reactor vessel in-situ decommissioning. Magnesium mono potassium phosphate cement-based grout was identified as candidate material for filling (physically stabilizing) the 105-P Reactor vessel (RV) because it is less alkaline than portland cement-based grout (pH of about 12.4). A less alkaline material ({<=} 10.5) was desired to address a potential materials compatibility issue caused by corrosion of aluminum metal in highly alkaline environments such as that encountered in portland cement grouts. Information concerning access points into the P-Reactor vessel and amount of aluminum metal in the vessel is provided elsewhere. Fresh and cured properties were measured for: (1) commercially blended magnesium mono potassium phosphate packaged grouts, (2) commercially available binders blended with inert fillers at SRNL, (3) grouts prepared from technical grade MgO and KH{sub 2}PO{sub 4} and inert fillers (quartz sands, Class F fly ash), and (4) Ceramicrete{reg_sign} magnesium mono potassium phosphate-based grouts prepared at Argonne National Laboratory. Boric acid was evaluated as a set retarder in the magnesium mono potassium phosphate mixes.

  7. High mitochondrial sequence diversity in linguistic isolates of the Alps.

    PubMed Central

    Stenico, M.; Nigro, L.; Bertorelle, G.; Calafell, F.; Capitanio, M.; Corrain, C.; Barbujani, G.

    1996-01-01

    Segment I of the control region of mtDNA (360 bases) was sequenced in seven samples, each of 10 individuals inhabiting villages in the eastern Italian Alps (South Tyrol and Trentino). Three linguistic groups, German, Italian, and Ladin, were represented by two samples each; the seventh sample comes from an isolated group of German origin, the Mocheni, who are linguistically distinct and geographically separated from the bulk of the German speakers. Seventy-four polymorphic sites were identified, defining 63 different haplotypes. Mocheni and Ladin speakers tend to form two clusters in the evolutionary trees inferred from sequences. Analysis of molecular variance shows significant differentiation within samples, among them, and among linguistic groups. Genetic differences between the Ladins and the other groups are not much smaller than between Europeans and some Africans; variation is large within groups, as well, with the exception of only the Mocheni. In the evolutionary trees where the four alpine groups are compared with other European populations, Mocheni and especially Ladins appear as clear outliers. Romansch-speaking Swiss, who are linguistically related to Ladins, are not genetically similar to them, for this segment of DNA. Because the time elapsed since colonization of the Alps (< or = 12,000 years) is short in mutational terms, the only model accounting for the observed relationships between mtDNA variation and linguistic identity seems one in which a population ancestral to Ladin speakers was already differentiated long before the Alps were settled and the current linguistic affiliations were established. For the Mocheni, the results are consistent with a simpler episode of allele loss, from an original genetic pool common to the ancestors of the current German speakers. PMID:8940282

  8. Fluorodeoxyglucose positron emission tomography (FDG-PET) for monitoring lymphadenopathy in the autoimmune lymphoproliferative syndrome (ALPS).

    PubMed

    Rao, V Koneti; Carrasquillo, Jorge A; Dale, Janet K; Bacharach, Stephen L; Whatley, Millie; Dugan, Faith; Tretler, Jean; Fleisher, Thomas; Puck, Jennifer M; Wilson, Wyndham; Jaffe, Elaine S; Avila, Nilo; Chen, Clara C; Straus, Stephen E

    2006-02-01

    Autoimmune lymphoproliferative syndrome (ALPS) is associated with mutations that impair the activity of lymphocyte apoptosis proteins, leading to chronic lymphadenopathy, hepatosplenomegaly, autoimmunity, and an increased risk of lymphoma. We investigated the utility of fluorodeoxyglucose positron emission tomography (FDG-PET) in discriminating benign from malignant lymphadenopathy in ALPS. We report that FDG avidity of benign lymph nodes in ALPS can be high and, hence, by itself does not imply presence of lymphoma; but FDG-PET can help guide the decision for selecting which of many enlarged nodes in ALPS patients to biopsy when lymphoma is suspected.

  9. Dephosphorylation of bovine casein by milk alkaline phosphatase.

    PubMed

    Lorient, D; Linden, G

    1976-02-01

    The pH of optimum activity of alkaline phosphatase from cow's milk depended on the substrate, being 10-1 for rho-nitrophenylphosphate, 8-6 for phosphoserine, 8-0 for phosvitin and 6-8 for casein. Individual casein components were dephosphorylated more rapidly than mixtures of alphas- and beta-caseins or of alphas-, beta-and kappa-caseins and micellar casein. Mixtures of 2 components involving kappa-casein were more readily dephosphorylated than alphas- and beta-casein mixtures. At pH 6-8, lactose, whey proteins and phosphate ions had an inhibitory effect. beta-Lactoglobulin had an inhibitory effect only when the pH of the reaction was lower than the optimum pH value of the enzyme. Mg2+ and Zn2+ were not inhibitory. The optimum conditions for dephosphorylation of casein are described.

  10. Improved Manganese Phosphate Coatings

    DTIC Science & Technology

    1975-04-01

    Conversion coatings 3 . Phosphating bath 20 AGrjC onln odd*. ta It .. c..soMV midP 1J.,alft. by block noc.mb) Work was conducted to determine the mechanism by...34 TABULAR DATA Table I Analyses of Solution and Coating for Phosphating Baths 4 of Di-ferlng Compositions 11 Atomic Absorption...manganese and iron phosphate coating: k * a. Mn(H 2PO4) 2 Nn-P0 4 + H3PO0 k2 k) b. 3MnHPO4 - Mn3 (P04) 2 + H3i’O4 k4 k5 c. Fe(H 2PO4) 2 -01 FeHPO4

  11. Phosphate Mines, Jordan

    NASA Technical Reports Server (NTRS)

    2008-01-01

    Jordan's leading industry and export commodities are phosphate and potash, ranked in the top three in the world. These are used to make fertilizer. The Jordan Phosphate Mines Company is the sole producer, having started operations in 1935. In addition to mining activities, the company produces phosphoric acid (for fertilizers, detergents, pharmaceuticals), diammonium phosphate (for fertilizer), sulphuric acid (many uses), and aluminum fluoride (a catalyst to make aluminum and magnesium).

    The image covers an area of 27.5 x 49.4 km, was acquired on September 17, 2005, and is located near 30.8 degrees north latitude, 36.1 degrees east longitude.

    The U.S. science team is located at NASA's Jet Propulsion Laboratory, Pasadena, Calif. The Terra mission is part of NASA's Science Mission Directorate.

  12. 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

  13. Glaciokarst: a Case Study from the Dinaric Alps

    NASA Astrophysics Data System (ADS)

    Žebre, Manja; Stepišnik, Uroš

    2014-05-01

    The Dinaric Alps are one of the largest contiguous mountain belts of the European part of Eurasian orogen with the total length of 645 km. The biggest part of the Dinaric Alps is formed of carbonate rocks, mostly limestone and dolomite. Here, the highest massifs are found and karst geomorphic system prevails. Well karstified Dinaric Alps were affected by the Quaternary glaciations. An interaction between glacial action and karst processes is most frequently termed as glaciokarst. Glaciokarst can function only if vertical karst drainage of subglacial waters is active, which results in specific functioning of glaciers as well as in formation of typical geomorphic features. On the other hand inert vertical karst drainage prevents functioning of glaciokarst and causes only subglacial and postglacial surface modification by dissolution, which is referred to as karsti-glacial action. The case study from the Lovćen, Orjen and Velež Mountains in the southern Dinaric Alps provides an insight into specific glaciokarst processes and surface features. Aside from variety of subglacial and postglacial dissolution karsti-glacial features, typical features of subglacial origin such as schafts, dolines, kotlic and kontas (over-deepened cirques) which are all the result of subglacial karstic drainage, are present in the study areas. Vertical karst drainage has also significant effect both on functioning of glaciers and on glacial deposition. Since prevailing process in karst areas following deglaciation is vertical chemical denudation, almost entire absence of other surface processes resulted in almost complete preservation of glacial accumulations. The dynamics of glaciers in karst areas is similar to the one in arid mountain areas where proglacial discharge is relatively low in comparison with sediment supply. Therefore, moraine-dammed glaciers are common in karst areas due to vertical drainage resulting in inability of ice-marginal fluvial processes to evacuate sediment. This

  14. Effects of Silicate, Phosphate, and Calcium on the Stability of Aldopentoses.

    PubMed

    Nitta, Sakiko; Furukawa, Yoshihiro; Kakegawa, Takeshi

    2016-06-01

    Ribose is an important constituent of RNA: ribose connects RNA bases and forms a strand of sugar phosphates. Accumulation of ribose on prebiotic Earth was difficult because of its low stability. Improvement in the yield of ribose by the introduction of borate or silicate in a formose-like reaction has been proposed. The effects of borates have been further analyzed and confirmed in subsequent studies. Nonetheless, the effects of silicates and phosphates remain unclear. In the present study, we incubated aldopentoses in a highly alkaline aqueous solution at a moderate temperature to determine the effects of silicate or phosphate on the degradation rates of ribose and its isomeric aldopentoses. The formation of a complex of silicate (or phosphate) with ribose was also analyzed in experiments with (29)Si and (31)P nuclear magnetic resonance (NMR). We found that silicate or phosphate complexes of ribose were not detectable under our experimental conditions. The stability of ribose and lyxose improved after addition of 40-fold molar excess (relative to a pentose) of sodium silicate or sodium phosphate to the alkaline solution. The stability was not improved further when an 80-fold molar excess of sodium silicate or sodium phosphate was added. Calcium was removed from these solutions by precipitation of calcium salts. The drop in Ca(2+) concentration might have improved the stability of ribose and lyxose, which are susceptible to aldol addition. The improvement of ribose stability by the removal of Ca(2+) and by addition of silicate or phosphate was far smaller than the improvement by borate. Furthermore, all aldopentoses showed similar stability in silicate- and phosphate-containing solutions. These results clearly show that selective stabilization of ribose by borate cannot be replaced by the effects of silicate or phosphate; this finding points to the importance of borate in prebiotic RNA formation.

  15. Effects of Silicate, Phosphate, and Calcium on the Stability of Aldopentoses

    NASA Astrophysics Data System (ADS)

    Nitta, Sakiko; Furukawa, Yoshihiro; Kakegawa, Takeshi

    2016-06-01

    Ribose is an important constituent of RNA: ribose connects RNA bases and forms a strand of sugar phosphates. Accumulation of ribose on prebiotic Earth was difficult because of its low stability. Improvement in the yield of ribose by the introduction of borate or silicate in a formose-like reaction has been proposed. The effects of borates have been further analyzed and confirmed in subsequent studies. Nonetheless, the effects of silicates and phosphates remain unclear. In the present study, we incubated aldopentoses in a highly alkaline aqueous solution at a moderate temperature to determine the effects of silicate or phosphate on the degradation rates of ribose and its isomeric aldopentoses. The formation of a complex of silicate (or phosphate) with ribose was also analyzed in experiments with 29Si and 31P nuclear magnetic resonance (NMR). We found that silicate or phosphate complexes of ribose were not detectable under our experimental conditions. The stability of ribose and lyxose improved after addition of 40-fold molar excess (relative to a pentose) of sodium silicate or sodium phosphate to the alkaline solution. The stability was not improved further when an 80-fold molar excess of sodium silicate or sodium phosphate was added. Calcium was removed from these solutions by precipitation of calcium salts. The drop in Ca2+ concentration might have improved the stability of ribose and lyxose, which are susceptible to aldol addition. The improvement of ribose stability by the removal of Ca2+ and by addition of silicate or phosphate was far smaller than the improvement by borate. Furthermore, all aldopentoses showed similar stability in silicate- and phosphate-containing solutions. These results clearly show that selective stabilization of ribose by borate cannot be replaced by the effects of silicate or phosphate; this finding points to the importance of borate in prebiotic RNA formation.

  16. Alkaline detergent recycling via ultrafiltration

    SciTech Connect

    Steffani, C.; Meltzer, M.

    1995-06-01

    The metal finishing industry uses alkaline cleaners and detergents to remove oils and dirt from manufactured parts, often before they are painted or plated. The use of these cleaners has grown because environmental regulations are phasing out ozone depleting substances and placing restrictions on the use and disposal of many hazardous solvents. Lawrence Livermore National Laboratory is examining ultrafiltration as a cleaning approach that reclaims the cleaning solutions and minimizes wastes. The ultrafiltration membrane is made from sheets of polymerized organic film. The sheets are rolled onto a supporting frame and installed in a tube. Spent cleaning solution is pumped into a filter chamber and filtered through the membrane that captures oils and dirt and allows water and detergent to pass. The membrane is monitored and when pressure builds from oil and dirt, an automatic system cleans the surface to maintain solution flow and filtration quality. The results show that the ultrafiltration does not disturb the detergent concentration or alkalinity but removed almost all the oils and dirt leaving the solution in condition to be reused.

  17. Enhanced Dentin-Like Mineralized Tissue Formation by AdShh-Transfected Human Dental Pulp Cells and Porous Calcium Phosphate Cement

    PubMed Central

    Chang, Qing; Wang, Lizhen; Zeng, Deliang; Zhang, Xiuli; Zhang, Zhiyuan; Jiang, Xinquan

    2013-01-01

    The aim of the present study was to investigate the effect of Sonic hedgehog (Shh) on human dental pulp cells (hDPCs) and the potential of complexes with Shh gene modified hDPCs and porous calcium phosphate cement (CPC) for mineralized tissue formation. hDPCs were cultured and transfected with adenoviral mediated human Shh gene (AdShh). Overexpression of Shh and cell proliferation was tested by real-time PCR analysis, western blotting analysis, and MTT analysis, respectively. The odontoblastic differentiation was assessed by alkaline phosphatase (ALP) activity and real-time PCR analysis on markers of Patched-1 (Ptc-1), Smoothened (Smo), Gli 1, Gli 2, Gli 3, osteocalcin (OCN), dentin matrix protein-1 (DMP-1), and dentin sialophosphoprotein (DSPP). Finally, AdShh-transfected hDPCs were combined with porous CPC and placed subcutaneously in nude mice for 8 and 12 weeks, while AdEGFP-transfected and untransfected hDPCs were treated as control groups. Results indicated that Shh could promote proliferation and odontoblastic differentiation of hDPCs, while Shh/Gli 1 signaling pathway played a key role in this process. Importantly, more mineralized tissue formation was observed in combination with AdShh transfected hDPCs and porous CPC, moreover, the mineralized tissue exhibited dentin-like features such as structures similar to dentin-pulp complex and the positive staining for DSPP protein similar to the tooth tissue. These results suggested that the constructs with AdShh-transfected hDPCs and porous CPC might be a better alternative for dental tissue regeneration. PMID:23675415

  18. Promiscuity in alkaline phosphatase superfamily. Unraveling evolution through molecular simulations.

    PubMed

    López-Canut, Violeta; Roca, Maite; Bertrán, Juan; Moliner, Vicent; Tuñón, Iñaki

    2011-08-10

    We here present a theoretical study of the alkaline hydrolysis of a phosphodiester (methyl p-nitrophenyl phosphate or MpNPP) in the active site of Escherichia coli alkaline phosphatase (AP), a monoesterase that also presents promiscuous activity as a diesterase. The analysis of our simulations, carried out by means of molecular dynamics (MD) simulations with hybrid quantum mechanics/molecular mechanics (QM/MM) potentials, shows that the reaction takes place through a D(N)A(N) or dissociative mechanism, the same mechanism employed by AP in the hydrolysis of monoesters. The promiscuous activity observed in this superfamily can be then explained on the basis of a conserved reaction mechanism. According to our simulations the specialization in the hydrolysis of phosphomonoesters or phosphodiesters, developed in different members of the superfamily, is a consequence of the interactions established between the protein and the oxygen atoms of the phosphate group and, in particular, with the oxygen atom that bears the additional alkyl group when the substrate is a diester. A water molecule, belonging to the coordination shell of the Mg(2+) ion, and residue Lys328 seem to play decisive roles stabilizing a phosphomonoester substrate, but the latter contributes to increase the energy barrier for the hydrolysis of phosphodiesters. Then, mutations affecting the nature or positioning of Lys328 lead to an increased diesterase activity in AP. Finally, the capacity of this enzymatic family to catalyze the reaction of phosphoesters having different leaving groups, or substrate promiscuity, is explained by the ability of the enzyme to stabilize different charge distributions in the leaving group using different interactions involving either one of the zinc centers or residues placed on the outer side of the catalytic site.

  19. Amorphous calcium phosphate and its application in dentistry.

    PubMed

    Zhao, Jie; Liu, Yu; Sun, Wei-Bin; Zhang, Hai

    2011-07-08

    Amorphous Calcium Phosphate (ACP) is an essential mineral phase formed in mineralized tissues and the first commercial product as artificial hydroxyapatite. ACP is unique among all forms of calcium phosphates in that it lacks long-range, periodic atomic scale order of crystalline calcium phosphates. The X-ray diffraction pattern is broad and diffuse with a maximum at 25 degree 2 theta, and no other different features compared with well-crystallized hydroxyapatite. Under electron microscopy, its morphological form is shown as small spheroidal particles in the scale of tenths nanometer. In aqueous media, ACP is easily transformed into crystalline phases such as octacalcium phosphate and apatite due to the growing of microcrystalline. It has been demonstrated that ACP has better osteoconductivity and biodegradability than tricalcium phosphate and hydroxyapatite in vivo. Moreover, it can increase alkaline phosphatase activities of mesoblasts, enhance cell proliferation and promote cell adhesion. The unique role of ACP during the formation of mineralized tissues makes it a promising candidate material for tissue repair and regeneration. ACP may also be a potential remineralizing agent in dental applications. Recently developed ACP-filled bioactive composites are believed to be effective anti-demineralizing/remineralizing agents for the preservation and repair of tooth structures. This review provides an overview of the development, structure, chemical composition, morphological characterization, phase transformation and biomedical application of ACP in dentistry.

  20. Pretreatment Alkaline Phosphatase and Epstein-Barr Virus DNA Predict Poor Prognosis and Response to Salvage Radiotherapy in Patients with Nasopharyngeal Carcinoma and Metachronous Bone-Only Metastasis

    PubMed Central

    He, ShaSha; Wang, Yan; Peng, Hao; Yang, Lin; Chen, HaiYang; Liang, ShaoBo; Lu, LiXia; Chen, Yong

    2017-01-01

    Background: The bones are the most common site of distant metastasis in nasopharyngeal carcinoma (NPC). Few prognostic markers are available to guide treatment and sub-classify patients with bone metastasis. We aimed to identify the prognostic value of pretreatment serum alkaline phosphatase (ALP) and plasma Epstein-Barr virus DNA (EBV DNA) in patients with bone-only metastasis. Methods: A total of 272 patients who developed bone-only metastases after therapy were retrospectively analyzed. Patients were categorized according to pretreatment serum ALP (< or ≥ 110 U/L) and pretreatment plasma EBV DNA (< or ≥ 6,750 copies ml-1). Univariate and multivariate analyses of clinical variables were performed using Cox proportional hazards regression models. Overall survival (OS) was analyzed using the Kaplan-Meier method and compared using the log-rank test. Results: Median OS for the cohort was 34.06 months (range, 2.53-143.87 months). Multivariate Cox proportional hazard analysis verified pretreatment serum ALP and pretreatment plasma EBV DNA were independent prognostic factors for OS. In stratified survival analysis of patients with elevated pretreatment serum ALP and/or plasma EBV DNA, delivery of radiotherapy (RT) to bone metastases provided a significant OS benefit compared to other therapeutic methods (P < 0.05). Conclusions: This study demonstrates two important points: firstly, pretreatment serum ALP and plasma EBV DNA have prognostic value at the first diagnosis of bone-only metastasis in NPC. Secondly, radiotherapy of bone metastasis improves the prognosis of patients with elevated pretreatment serum ALP and plasma EBV DNA. PMID:28261343

  1. A new phosphorimetry for the determination of trace alkaline phosphatase using multi-wall carbon nanotubes and its diagnosis of human diseases.

    PubMed

    Liu, Jia-Ming; Rao, Ying; Chen, Li; Wang, Xin-Xing; Lin, Li-Ping; Lin, Chang-Qing; Zhang, Li-Hong; Ou-Yang, Ying

    2010-07-01

    Multi-wall carbon nanotubes (MWNTs) could be modified as water soluble MWNTs (it was called as MWNTs-A and MWNTs-B) by chemical methods. MWNTs-A and MWNTs-B could emit room temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM). A new solid substrate-room temperature phosphorimetry (SS-RTP) for the determination of trace alkaline phosphatase (ALP) was established based on the signal magnification effect of tween-80 and ALP on MWNTs-B's RTP intensity and the linear relationship between the content of ALP and the DeltaI(P) of the system. The linear range of this method was 0.0020-0.80 (fg spot(-1), sample volume: 0.40 microl spot(-1)), the regression equation of working curve was DeltaI(P) = 0.8170 + 96.84m(ALP) (fg spot(-1)), correlation coefficient (r) was 0.9986. This method had high sensitivity (detection limit (LD): 1.4 ag spot(-1)), good selectivity (Er < or = +/- 5 in-care-of, coexistence species were of no interference), high precision (RSDs were 4.4%-1.2%) and accuracy. It was applied to the determination of trace ALP in human serum and the diagnosis of human diseases. The results were tallied with those of enzyme-linked immunosorbent assay (ELISA). The mechanism of new SS-RTP for the determination of trace ALP was discussed, which laid the theory foundation for the analytical application of MWNTs in life science.

  2. 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.

  3. 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.

  4. Alkaline volcanic rocks from the Columbia Hills, Gusev crater, Mars

    NASA Astrophysics Data System (ADS)

    McSween, H. Y.; Ruff, S. W.; Morris, R. V.; Bell, J. F.; Herkenhoff, K.; Gellert, R.; Stockstill, K. R.; Tornabene, L. L.; Squyres, S. W.; Crisp, J. A.; Christensen, P. R.; McCoy, T. J.; Mittlefehldt, D. W.; Schmidt, M.

    2006-09-01

    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.

  5. Distributed landsurface skin temperature sensing in Swiss Alps

    NASA Astrophysics Data System (ADS)

    van de Giesen, N.; Baerenbold, F.; Nadeau, D. F.; Pardyjak, E.; Parlange, M. B.

    2010-12-01

    The ZyTemp TN9 is a mass-produced thermal infrared (TIR) sensor that is normally used to build handheld non-contact thermometers. The measurement principle of the TN9 is similar to that of very costly meteorological pyrgeometers. The costs of the TN9 are less than 10. The output of the TN9 consists of observed thermal radiation, the temperature of the measurement instrument, and the emissivity used. The output is provided through a Serial Peripheral Interface protocol. The TN9 was combined with an Arduino board that registered data onto a USB memory stick. A solar cell, lead acid battery, housing and stand completed the meausrement set up. Total costs per set was in the order of 200 Land surface atmosphere interactions in mountainous areas, such as the Swiss Alps, are spatially heterogeneous. Shading, multi-layer cloud formation, and up- and downdrafts make for a very dynamic exchange of mass and energy along and across slopes. In order to better understand these exchanges, the Swiss Slope Experiment at La Fouly (SELF) has built a distributed sensing network consisting of eight micro-met stations and two flux towers in the "La Fouly" watershed in the upper Alps. To obtain a better handle on surface temperature, fifteen TIR sensing stations were installed that made observations during the 2010 Summer. Methods and results will be presented. Overview La Fouly watershed (source: http://eflum.epfl.ch/research/images/fouly_2.jpg)

  6. 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.

  7. Osseous plate alkaline phosphatase is anchored by GPI.

    PubMed

    Pizauro, J M; Ciancaglini, P; Leone, F A

    1994-02-01

    Alkaline phosphatase activity was released up to 100% from the membrane by using 0.1 U of phosphatidylinositol-specific phospholipase C from B. thuringiensis. The M(r) of solubilized enzyme was 145,000 by Sephacryl S-300 gel filtration and 66,000 by SDS-PAGE, suggesting a dimeric structure. Solubilization of the membrane-bound enzyme with phospholipase C did not destroy its ability to hydrolyze p-nitrophenyl phosphate (PNPP) (264.3 mumol min-1 mg-1),ATP (42.0 mumol min-1 mg-1) and pyrophosphate (28.4 mumol min-1 mg-1). The hydrolysis of ATP and PNPP by solubilized enzyme exhibited "Michaelian" kinetics with K0.5 = 70 and 979 microM, respectively. For pyrophosphate, K0.5 was 128 microM and site-site interactions were observed (n = 1.4). Magnesium ions were stimulatory (Kd = 1.5 mM) but zinc ions were powerful non-competitive inhibitors (Kd = 6.2 microM) of solubilized enzyme. Treatment of solubilized alkaline phosphatase with Chellex 100 reduced the original PNPPase activity to 5%. Cobalt (K0.5 = 10.1 microM), magnesium (K0.5 = 29.5 microM) and manganese ions (K0.5 = 5 microM) restored the activity of the apoenzyme with positive cooperativity, suggesting that phosphatidylinositol-specific phospholipase C-solubilized alkaline phosphatase is a metalloenzyme. The stimulation of the apoenzyme by calcium ions (K0.5 = 653 microM) was lower than that observed for the other ions (26%) and exhibited site-site interactions (n = 0.7). Zinc ions had no effect on the apoenzyme of the solubilized enzyme.

  8. Domestic phosphate deposits

    USGS Publications Warehouse

    McKelvey, V.E.; Cathcart, J.B.; Altschuler, Z.S.; Swanson, R.W.; Lutz, Katherine

    1953-01-01

    Most of the worlds phosphate deposits can be grouped into six types: 1) igneous apatite deposits; 2) marine phosphorites; 3) residual phosphorites; 4) river pebble deposits; 5) phosphatized rock; and 6) guano. The igneous apatites and marine phosphorites form deposits measurable in millions or billions of tons; the residual deposits are measurable in thousands or millions; and the other types generally only in thousands of tons. Igneous apatite deposits have been mined on a small scale in New York, New Jersey, and Virginia. Marine phosphorites have been mined in Montana, Idaho, Utah, Wyoming, Arkansas, Tennessee, North Carolina, South Carolina, Georgia, and Florida. Residual phosphorites have been mined in Tennessee, Pennsylvania, and Florida. River pebble has been produced in South Carolina and Florida; phosphatized rock in Tennessee and Florida; and guano in New Mexico and Texas. Present production is limited almost entirely to Florida, Tennessee, Montana, Idaho, and Wyoming. Incomplete but recently partly revised estimates indicate the presence of about 5 billion tons of phosphate deposits in the United States that is minable under present economic conditions. Deposits too lean in quality or thickness to compete with those in the western and southeastern fields probably contain tens of billions of tons.

  9. Glucose-6-phosphate dehydrogenase

    MedlinePlus

    ... Elsevier Saunders; 2012:chap 42. Read More Enzyme Glucose-6-phosphate dehydrogenase deficiency Hemoglobin Review Date 2/11/2016 Updated by: ... A.M. Editorial team. Related MedlinePlus Health Topics G6PD Deficiency Browse the Encyclopedia A.D.A.M., Inc. ...

  10. 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

  11. 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.

  12. Molecular defect of tissue-nonspecific alkaline phosphatase bearing a substitution at position 426 associated with hypophosphatasia.

    PubMed

    Al-Shawafi, Hiba A; Komaru, Keiichi; Oda, Kimimitsu

    2017-03-01

    Mutations in the ALPL gene encoding tissue-nonspecific alkaline phosphatase (TNSALP) cause hypophosphatasia (HPP), a genetic disorder characterized by deficiency of serum ALP and hypomineralization of bone and teeth. Three missense mutations for glycine 426 (by standard nomenclature) of TNSALP have been reported: cysteine (p.G426C), serine (p.G426S), and aspartate (p.G426D). We expressed TNSALP mutants carrying each missense mutation in mammalian cells. All three TNSALP mutants appeared on the cell surface like the wild-type (WT) TNSALP, although the cells expressing each TNSALP mutant exhibited markedly reduced ALP activity. TNSALP (WT) was mainly present as a 140 kDa catalytically active dimeric form, whereas ~80 kDa monomers were the predominant molecular species in the cells expressing TNSALP (p.G426D) or TNSALP (p.G426S), suggesting that aspartate or serine at position 426 may hamper the subunit assembly essential for the enzymatic function of TNSALP. Alternatively, the subunits of TNSALP (p.G426C) were found to be aberrantly cross-linked by disulfide bonds, giving rise to a 200 kDa form lacking ALP activity. Taken together, our results reveal that the amino acid substitutions at position 426 of TNSALP differentially affect the structure and function of TNSALP, leading to understanding of the molecular and cellular basis of HPP.

  13. Panax notoginseng stimulates alkaline phosphatase activity, collagen synthesis, and mineralization in osteoblastic MC3T3-E1 cells.

    PubMed

    Ji, Zhe; Cheng, Yizhao; Yuan, Puwei; Dang, Xiaoqian; Guo, Xiong; Wang, Weizhuo

    2015-10-01

    Total Panax notoginseng saponin (PNS) has been extensively used to treat a variety of diseases, such as bone fractures, soft tissue injuries, etc. In this study, mouse calvaria-original osteoblastic MC3T3-E1 cells were cultured in various concentrations of PNS (0.005-5 mg/mL) during the period (1, 5, 14, and 23 d). At the endpoint, the osteogenic capacity of MC3T3-E1 cells was investigated by measuring the alkaline phosphatase (ALP) activity, the deposited calcium, and the expression of osteogenic-related markers, including bone collagen type 1 (Col1) and osteocalcin (OCN). Compared with all groups in each period, the most pronounced effect was observed at the concentration range between 0.05 and 0.5 mg/mL (P < 0.05) and the cell proliferation with PNS treatment was found during the whole osteogenic period. Moreover, cellular ALP activity with PNS was increased during 7, 14, and 21 d and cell mineralization with PNS was enhanced in 14 and 21 d. Furthermore, the differentiation markers Col1 and OCN increased in the PNS-treated cells. Our work suggests that PNS may stimulate the osteogenesis process which contains osteoblastic proliferation, differentiation, and mineralization by increasing cellular ALP activity, extracellular matrix mineralization, and osteoblast-associated molecules in the osteoblasts.

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

    PubMed

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

    2014-06-01

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

  15. Comparative studies with penicillinase, horseradish peroxidase, and alkaline phosphatase as enzyme labels in developing enzyme immunoassay of cortisol.

    PubMed

    Kumari, G Lakshmi; Dhir, Ravindra N

    2003-01-01

    Relative merit of different enzyme labels for measuring cortisol directly in serum by competitive enzyme immunoassay (ELISA) was examined. Cortisol-21-hemisuccinate was labeled separately with penicillinase, horseradish peroxidase (HRP), and alkaline phosphatase (ALP) under identical reaction conditions. Antibody developed in rabbits against cortisol-3-0-(carboxymethyl)-oxime-bovine serum albumin was used to coat polystyrene tubes that were precoated with anti-rabbit gamma globulin (ARGG). Cortisol standards were prepared in steroid-free human serum in buffer (1:4) contaning 8-anilino-1-naphthalene sulfonic acid (8-ANS). Assay buffer also consisted 8-ANS. The assay involved adding standard cortisol or serum sample to antibody-coated tubes, followed by addition of enzyme label and buffer, and incubation for 2 h at 37 degrees C. The whole procedure took 3 h for completion. All three labels proved to be sensitive, with a slope around -2.0. Although penicillinase as an enzyme label was highly sensitive and stable compared with others, the assays were not always accurate and precise, especially at low concentrations of cortisol. This was mainly due to the color reagent used for measuring penicillinase activity. Serum samples that underwent 2-3 freeze-thaw cycles gave high values with HRP label compared with ALP. Therefore, utilizing ALP as an enzyme label, an ELISA was developed and its performance was comparable with some of the commercial kits already in the market.

  16. Evaluation of Manganese Phosphate Coatings.

    DTIC Science & Technology

    1984-02-01

    84003 _____________ 4 . TTLE and -bitle)5. TYPE OF REPORT & PERIOD COVERED EVALUATION OF MANGANESE PHOSPHATE COATINGS Final 6. PERFORMING ORG. REPORT...rosion resistance of the Endurion phosphate was significantly superior to the 4 . basic manganese phosphate . Endurion phosphate with a Supplementary...OF CONTENTS Page STATEMENT OF THE PROBLEM 1 BACKGROUND 1 APPROACH TO THE PROBLEM 3 RESULTS 4 CONCLUSIONS 7 TABLES I. Falex Wear Life Test Procedure 8

  17. Closed type alkaline storage battery

    SciTech Connect

    Hayama, H.

    1980-06-10

    The alkaline storage battery employs a metallic hat shaped terminal closure which has a piercing needle as well as a puncturable metallic diaphragm positioned below the piercing needle. The needle is fixed by caulking at its peripheral edge portion to a edge of the closure. A comparatively thick and hard metal plate is placed on the inner surface of the diaphragm and is applied to an open portion of a tubular metallic container which has a battery element. A peripheral edge portion of the closure, the diaphragm and the metallic plate are clamped in airtight relationship through a packing between the caulked end portion and an inner annular step portion of the metallic container of the battery. A lead wire extends from one polarity electrode of the battery element and is connected to a central portion of the metallic plate.

  18. [Detection of AlpA and AlpB lytic endopeptidase propeptides of Lysobacter sp. XL1 by sandwich-enzyme immunoassay based on monoclonal antibodies].

    PubMed

    Rudenko, N V; Tsfasman, I M; Latypov, O R; Ledova, L A; Krasovskaia, L A; Karatovskaia, A P; Brovko, F A; Vasil'eva, N V; Stepnaia, O A

    2014-01-01

    The extracellular lytic endopeptidases AlpA and AlpB of the bacterium Lysobacter sp. XL1 are highly homologous and synthesized as precursors consisting of signal peptide, propeptide and mature form. In this work, two monoclonal antibodies against propeptide endopeptidase AlpA (ProA) and eleven against propeptide endopeptidase AlpB (ProB) were obtained to study the AlpA and AlpB endopeptidases secretion. The affinity constants of the antibodies against ProA were 2.9 x 10(9) and 3.5 x 10(9) M(-1), and the affinity constants of the antibodies against ProB were from 1.5 x 10(8) to 2.2 x 10(9) M(-1). The obtained antibodies did not have cross-reactivity between themselves, as well as mature forms of the enzymes. The monoclonal antibodies based sandwich-type enzyme immunoassay has been developed for measuring the propeptide in a native form. The linear range of determination ProA was 1.5-100 ng/mL with 6% error of measurement, and for determining ProB 0.2-6.25 ng/mL with 6% error. Using the developed assay, ProA and ProB propeptides have been detected in cell lysates of Lysobacter sp. XL1 in an amount 1.18 ± 0.03 ng and 0.096 ± 0.002 ng per 1 OD540 of the bacterial culture, respectively. The immunochemical assay for detection various forms of AlpA and AlpB lytic endopeptidases can be useful when dealing with issues related to their secretion into the environment.

  19. A malachite green procedure for orthophosphate determination and its use in alkaline phosphatase-based enzyme immunoassay.

    PubMed

    Baykov, A A; Evtushenko, O A; Avaeva, S M

    1988-06-01

    An improved procedure for phosphate determination based on a highly colored complex of phosphomolybdate and malachite green is described. All necessary reagents are combined in one concentrated solution, making the assay sensitive and convenient. The procedure is based on the finding that the dye is easily soluble and stable in the presence of 6 N acid. The addition of Tween 20 is required to stabilize the dye-phosphomolybdate complex at phosphate concentrations above 10 microM. The time of color development at 25 degrees C is about 3 min. The procedure was adopted to measure alkaline phosphate activity in heterogeneous enzyme immunoassay with rho-nitrophenyl phosphate and pyrophosphate as substrates. In both cases, a 4-fold increase in sensitivity in terms of absorbance readings was obtained compared to the standard method based on rho-nitrophenol measurement. In visual analysis, the gain in sensitivity was as high as 20-fold, due to contrast color change (yellow to greenish blue).

  20. Impact of Accelerated Learning Program (ALP) and Other Assistance, 1999-2000.

    ERIC Educational Resources Information Center

    Baenen, Nancy; Lloyd, Wanda

    The Accelerated Learning Program (ALP) was a major new initiative in the Wake County Public School System (WCPSS), North Carolina, in 2000. The ALP was designed to help WCPSS meet its achievement goal of 95% of students scoring at or above grade level at grades 3 and 8 by 2003, with grade levels determined by the North Carolina End of Grade tests.…

  1. Unmasking Evans syndrome: T-cell phenotype and apoptotic response reveal autoimmune lymphoproliferative syndrome (ALPS).

    PubMed

    Teachey, David T; Manno, Catherine S; Axsom, Kelly M; Andrews, Timothy; Choi, John K; Greenbaum, Barbara H; McMann, Joseph M; Sullivan, Kathleen E; Travis, Susan F; Grupp, Stephan A

    2005-03-15

    Autoimmune lymphoproliferative syndrome (ALPS) is a rare disorder of disrupted lymphocyte homeostasis. Clinical manifestations of ALPS vary but typically include autoimmune cytopenias, organomegaly, lymphadenopathy, and increased risk of malignancies. A similar spectrum of symptoms may be seen in some patients with Evans syndrome (ES), a hematologic disorder defined by autoimmune destruction of at least 2 hematologic cell types. We hypothesized that a subset of patients diagnosed with ES may have ALPS. We screened 12 children with ES by flow cytometric analysis for CD4-/CD8- (double negative) T cells (DNTs) and with the definitive test for ALPS, defective in vitro Fas-mediated apoptosis. Six of the patients had elevated DNTs, suggestive of ALPS and also had defective Fas-mediated apoptosis. The other 6 patients displayed normal T-cell apoptosis; 5 of whom had normal DNTs, and 1 had a borderline result. Thus, 7 (58%) of 12 patients with ES had elevated DNTs suggestive of ALPS, with functional confirmation in 6 of 7. This suggests that analysis of DNTs may be a sensitive first-line screening test, serving as a marker of patients who should undergo definitive testing for ALPS. Our data further suggest that a number of patients with ES may have ALPS, a novel finding with important therapeutic implications.

  2. ALP: Alternate Learning Project; Overview of a Model High School in Providence, Rhode Island.

    ERIC Educational Resources Information Center

    Kenyon, Charles B.

    The Alternate Learning Project (ALP) is a community based public high school in Providence, Rhode Island. The ALP student population participates in a program offering individualized basic skills instruction, college preparatory courses, career exploration activities, and a broad arts curriculum. Throughout, the emphasis is on continuous…

  3. Evolution of alkaline phosphatases in primates.

    PubMed Central

    Goldstein, D J; Rogers, C; Harris, H

    1982-01-01

    Alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] in placenta, intestine, liver, kidney, bone, and lung from a variety of primate species has been characterized by quantitative inhibition, thermostability, and immunological studies. Characteristic human placental-type alkaline phosphatase occurs in placentas of great apes (chimpanzee and orangutan) but not in placentas of other primates, including gibbon. It is also present in trace amounts in human lung but not in lung or other tissues of various Old and New World monkeys. However, a distinctive alkaline phosphatase resembling it occurs in substantial amounts in lungs from Old World monkeys but not New World monkeys. It appears that duplication of alkaline phosphatase genes and mutations of genetic elements controlling their tissue expression have occurred relatively recently in mammalian evolution. Images PMID:6950431

  4. 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.

  5. Green synthesis of a novel hybrid sorbent of zeolite/lanthanum hydroxide and its application in the removal and recovery of phosphate from water.

    PubMed

    Xie, Jie; Wang, Zhe; Fang, Da; Li, Chunjie; Wu, Deyi

    2014-06-01

    The traditional process of zeolite synthesis from coal fly ash produces large amount of waste alkaline solution, besides zeolite product. A novel hybrid sorbent, which was composed of zeolite and lanthanum hydroxide (La-ZFA), was produced by using soluble lanthanum chloride to react with waste alkaline solution after the traditional process. This study investigated the capability of phosphate removal by La-ZFA. The sorbent has a high phosphate removal capacity, with a sorption maximum of 71.94 mg/g, according to the Langmuir model. The removal of phosphate by La-ZFA performs well at a wide pH range, reaching>95% from pH 2.5 to pH 10.5 when initial P concentration<100 mg/L. Release of OH(-) during the interaction of phosphate with La-ZFA was observed. The formation of the monoclinic LaPO4 phase was identified by XRD analysis. The uptake of phosphate by La-ZFA was explained on the basis of the adsorption mechanism of the ligand exchange process. The sorbed phosphate could be recovered by hydrothermal treatment in 3M NaOH at 250°C, with a simultaneous regeneration of La-ZFA. Repeated sorption-desorption tests indicated that regenerated La-ZFA could be reused for phosphate removal. Therefore, synthesis of La-ZFA could impart additional function of phosphate removal to original zeolite, with the effective utilization of waste alkaline solution.

  6. 21st Century Climate Change in the European Alps

    NASA Astrophysics Data System (ADS)

    Gobiet, Andreas; Kotlarski, Sven; Stoffel, Markus; Heinrich, Georg; Rajczak, Jan; Beniston, Martin

    2014-05-01

    The Alps are particularly sensitive to global warming and warmed twice as much as the global average in the recent past. In addition, the Alps and its surroundings are a densly populated areas where society is affected by climate change in many ways, which calls for reliable estimates of future climate change. However, the complex Alpine region poses considerable challenges to climate models, which translate to uncertainties in future climate projections. Against this background, the present study reviews the state-of-knowledge about 21st century climate change in the Alps based on existing literature and additional analyses. It will be demonstrated that considerable and accelerating changes are not only to be expected with regard to temperature, but also precipitation, global radiation, relative humidity, and closely related impacts like floods, droughts, snow cover, and natural hazards will be effected by global warming. Under the A1B emission scenario, about 0.25 °C warming per decade until the mid of the 21st century and accelerated 0.36 °C warming per decade in the second half of the century is expected. Warming will most probably be associated with changes in the seasonality of precipitation, global radiation, and relative humidity. More intense precipitation extremes and flooding potential are particularly expected in the colder part of the year. The conditions of currently record breaking warm or hot winter or summer seasons, respectively, may become normal at the end of the 21st century, and there is indication for droughts to become more severe in the future. Snow cover is expected to drastically decrease below 1500 - 2000 m and natural hazards related to glacier and permafrost retreat are expected to become more frequent. Such changes in climatic variables and related quantities will have considerable impact on ecosystems and society and will challenge their adaptive capabilities. Acknowledgements: This study has been initiated and is partly funded by

  7. Quantification of the seismic strain in the western Alps

    NASA Astrophysics Data System (ADS)

    Sue, C.; Delacou, B.; Champagnac, J. D.; Burkhard, M.

    2003-04-01

    The western alpine tectonic regime is characterized by ongoing widespread extension in the highest zones of the belt and transpressive/compressive tectonics at the external limits of the belt [Sue et al., 1999, Delacou et al, submitted]. The overall geodetic strain corresponds also to a radial extension across the western Alps [Vigny et al., 2001] and the data of the permanent GPS networks in western Europe provided an alpine strain pattern pretty close to the strain/stress states inferred from seismotectonics [Calais et al., 2002]. Thus, there is a quite good qualitative coherency between seismotectonic and geodetic approaches. We attempt here to quantify the seismic part of the deformation. The aim is to compare the seismic strain with the overall geodetic strain, at the local and global scales. We used the regionalization of the seismic deformation and the stress inversion of Delacou et al. [submitted] to determine sub-areas of quite homogeneous seismic strain. In each sub-area, we computed the total seismic moment tensor, and the associated yearly seismic strain rate. This rate allows to compare different areas of the belt, in which the seismic catalogue may cover various range of time. Thus we can estimate the seismic strain per sub-area in the whole belt. To improve this approach, we use typical magnitude distributions (b-values) in large areas of the belt [e.g. Sue et al., 2002] to take into account the part of the deformation which corresponds to the smaller earthquakes (without focal mechanisms). A second step of improvement will be to take into account historical catalogues available in the Alps to derive the seismic strain on larger time scale, and to compare the seismic strains of the historical vs. the instrumental periods. This preliminary study brings new quantitative elements to understand the ongoing geodynamic processes in the alpine belt. The low seismic strain rates we obtain in a belt characterized by a high tectonic contrast in a quite limited

  8. 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

  9. Risk assessment of mountain infrastructure destabilization in the French Alps

    NASA Astrophysics Data System (ADS)

    Duvillard, Pierre-Allain; Ravanel, Ludovic; Deline, Philip

    2015-04-01

    In the current context of imbalance of geosystems in connection with the rising air temperature for several decades, high mountain environments are especially affected by the shrinkage of glaciers and the permafrost degradation which can trigger slope movements in the rock slopes (rockfall, rock avalanches) or in superficial deposits (slides, rock glacier rupture, thermokarst). These processes generate a risk of direct destabilization for high mountain infrastructure (huts, cable-cars...) in addition to indirect risks for people and infrastructure located on the path of moving rock masses. We here focus on the direct risk of infrastructure destabilization due to permafrost degradation and/or glacier shrinkage in the French Alps. To help preventing these risks, an inventory of all the infrastructure was carried out with a GIS using different data layers among which the Alpine Permafrost Index Map and inventories of the French Alps glaciers in 2006-2009, 1967-1971 and at the end of the Little Ice Age. 1769 infrastructures have been identified in areas likely characterized by permafrost and/or possibly affected by glacier shrinkage. An index of risk of destabilization has been built to identify and to rank infrastructure at risk. This theoretical risk index includes a characterization of hazards and a diagnosis of the vulnerability. The value of hazard is dependent on passive factors (topography, lithology, geomorphological context...) and on so-considered active factors (thermal state of the permafrost, and changing constraints on slopes related to glacier shrinkage). The diagnosis of vulnerability has meanwhile been established by combining the level of potential damage to the exposed elements with their operational and financial values. The combination of hazard and vulnerability determines a degree of risk of infrastructure destabilization (from low to very high). Field work and several inventories of infrastructure damages were used to validate it. The

  10. 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

  11. Final Report: Dominant Mechanisms of Uranium-Phosphate Reactions in Subsurface Sediments

    SciTech Connect

    Catalano, Jeffrey G.; Giammar, Daniel E.; Wang, Zheming

    2016-03-08

    Phosphate addition is an in situ remediation approach that may enhance the sequestration of uranium without requiring sustained reducing conditions. However, the geochemical factors that determine the dominant immobilization mechanisms upon phosphate addition are insufficiently understood to design efficient remediation strategies or accurately predict U(VI) transport. The overall objective of our project is to determine the dominant mechanisms of U(VI)-phosphate reactions in subsurface environments. Our research approach seeks to determine the U(VI)-phosphate solid that form in the presence of different groundwater cations, characterize the effects of phosphate on U(VI) adsorption and precipitation on smectite and iron oxide minerals, examples of two major reactive mineral phases in contaminated sediments, and investigate how phosphate affects U(VI) speciation and fate during water flow through sediments from contaminated sites. The research activities conducted for this project have generated a series of major findings. U(VI) phosphate solids from the autunite mineral family are the sole phases to form during precipitation, with uranyl orthophosphate not occurring despite its predicted greater stability. Calcium phosphates may take up substantial quantities of U(VI) through three different removal processes (adsorption, coprecipitation, and precipitation) but the dominance of each process varies with the pathway of reaction. Phosphate co-adsorbs with U(VI) onto smectite mineral surfaces, forming a mixed uranium-phosphate surface complex over a wide range of conditions. However, this molecular-scale association of uranium and phosphate has not effect on the overall extent of uptake. In contrast, phosphate enhanced U(VI) adsorption to iron oxide minerals at acidic pH conditions but suppresses such adsorption at neutral and alkaline pH, despite forming mixed uranium-phosphate surface complexes during adsorption. Nucleation barriers exist that inhibit U(VI) phosphate

  12. Calcium Phosphates and Human Beings

    NASA Astrophysics Data System (ADS)

    Dorozhkin, Sergey V.

    2006-05-01

    This article describes the general importance of calcium phosphates for human beings. The basic information on the structure and chemical properties of the biologically relevant calcium phosphates is summarized. Basic facts on the natural occurrence and the industrial use of natural calcium phosphates are discussed. Fundamental details on the presence of calcium phosphates in major calcified tissues (bones and teeth) of humans and mammals, as well as on biomaterials made of calcium phosphates are discussed. The article will be of value for chemistry teachers for expansion of their general background and point the students' attention to the rapidly growing topic of bone-substituting biomaterials.

  13. 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,...

  14. 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,...

  15. 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....

  16. 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.

  17. 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.

  18. 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.

  19. Climate change and geomorphological hazards in the eastern European Alps.

    PubMed

    Keiler, Margreth; Knight, Jasper; Harrison, Stephan

    2010-05-28

    Climate and environmental changes associated with anthropogenic global warming are being increasingly identified in the European Alps, as seen by changes in long-term high-alpine temperature, precipitation, glacier cover and permafrost. In turn, these changes impact on land-surface stability, and lead to increased frequency and magnitude of natural mountain hazards, including rock falls, debris flows, landslides, avalanches and floods. These hazards also impact on infrastructure, and socio-economic and cultural activities in mountain regions. This paper presents two case studies (2003 heatwave, 2005 floods) that demonstrate some of the interlinkages between physical processes and human activity in climatically sensitive alpine regions that are responding to ongoing climate change. Based on this evidence, we outline future implications of climate change on mountain environments and its impact on hazards and hazard management in paraglacial mountain systems.

  20. AlpArray-Italy: Site description and noise characterization

    NASA Astrophysics Data System (ADS)

    Govoni, Aladino; Bonatto, Luciana; Capello, Marco; Cavaliere, Adriano; Chiarabba, Claudio; D'Alema, Ezio; Danesi, Stefania; Lovati, Sara; Margheriti, Lucia; Massa, Marco; Mazza, Salvatore; Mazzarini, Francesco; Monna, Stephen; Moretti, Milena; Nardi, Anna; Piccinini, Davide; Piromallo, Claudia; Pondrelli, Silvia; Salimbeni, Simone; Serpelloni, Enrico; Solarino, Stefano; Vallocchia, Massimiliano; Santulin, Marco; AlpArray Working Group

    2017-03-01

    Within the framework of the European collaborative research initiative AlpArray (http://www.alparray.ethz.ch), the Istituto Nazionale di Geofisica e Vulcanolgia (INGV) deployed overall 20 broad-band seismic stations in Northern Italy and on two islands in the Tyrrhenian Sea (Capraia and Montecristo) during Fall-Winter 2015. The temporary deployment (16 stations) will run for two to three years and 4 INGV National Seismic Network accelerometric sites are now equipped with additional permanent broad-band sensors. The 16 temporary stations are equipped with REF TEK 130 digitizers and Nanometrics Trillium Compact 120 s sensors, a couple have Nanometrics Trillium 120P sensors and one a Streckeisen STS2. For each site we describe the settings and discuss the noise levels, the site effects and the preliminary sensitivity analysis.

  1. Renal phosphate handling: Physiology

    PubMed Central

    Prasad, Narayan; Bhadauria, Dharmendra

    2013-01-01

    Phosphorus is a common anion. It plays an important role in energy generation. Renal phosphate handling is regulated by three organs parathyroid, kidney and bone through feedback loops. These counter regulatory loops also regulate intestinal absorption and thus maintain serum phosphorus concentration in physiologic range. The parathyroid hormone, vitamin D, Fibrogenic growth factor 23 (FGF23) and klotho coreceptor are the key regulators of phosphorus balance in body. PMID:23961477

  2. Crystal structures of Bacillus alkaline phytase in complex with divalent metal ions and inositol hexasulfate.

    PubMed

    Zeng, Yi-Fang; Ko, Tzu-Ping; Lai, Hui-Lin; Cheng, Ya-Shan; Wu, Tzu-Hui; Ma, Yanhe; Chen, Chun-Chi; Yang, Chii-Shen; Cheng, Kuo-Joan; Huang, Chun-Hsiang; Guo, Rey-Ting; Liu, Je-Ruei

    2011-06-03

    Alkaline phytases from Bacillus species, which hydrolyze phytate to less phosphorylated myo-inositols and inorganic phosphate, have great potential as additives to animal feed. The thermostability and neutral optimum pH of Bacillus phytase are attributed largely to the presence of calcium ions. Nonetheless, no report has demonstrated directly how the metal ions coordinate phytase and its substrate to facilitate the catalytic reaction. In this study, the interactions between a phytate analog (myo-inositol hexasulfate) and divalent metal ions in Bacillus subtilis phytase were revealed by the crystal structure at 1.25 Å resolution. We found all, except the first, sulfates on the substrate analog have direct or indirect interactions with amino acid residues in the enzyme active site. The structures also unraveled two active site-associated metal ions that were not explored in earlier studies. Significantly, one metal ion could be crucial to substrate binding. In addition, binding of the fourth sulfate of the substrate analog to the active site appears to be stronger than that of the others. These results indicate that alkaline phytase starts by cleaving the fourth phosphate, instead of the third or the sixth that were proposed earlier. Our high-resolution, structural representation of Bacillus phytase in complex with a substrate analog and divalent metal ions provides new insight into the catalytic mechanism of alkaline phytases in general.

  3. Exposure to the proton scavenger glycine under alkaline conditions induces Escherichia coli viability loss.

    PubMed

    Vanhauteghem, Donna; Janssens, Geert Paul Jules; Lauwaerts, Angelo; Sys, Stanislas; Boyen, Filip; Cox, Eric; Meyer, Evelyne

    2013-01-01

    Our previous work described a clear loss of Escherichia coli (E. coli) membrane integrity after incubation with glycine or its N-methylated derivatives N-methylglycine (sarcosine) and N,N-dimethylglycine (DMG), but not N,N,N-trimethylglycine (betaine), under alkaline stress conditions. The current study offers a thorough viability analysis, based on a combination of real-time physiological techniques, of E. coli exposed to glycine and its N-methylated derivatives at alkaline pH. Flow cytometry was applied to assess various physiological parameters such as membrane permeability, esterase activity, respiratory activity and membrane potential. ATP and inorganic phosphate concentrations were also determined. Membrane damage was confirmed through the measurement of nucleic acid leakage. Results further showed no loss of esterase or respiratory activity, while an instant and significant decrease in the ATP concentration occurred upon exposure to either glycine, sarcosine or DMG, but not betaine. There was a clear membrane hyperpolarization as well as a significant increase in cellular inorganic phosphate concentration. Based on these results, we suggest that the inability to sustain an adequate level of ATP combined with a decrease in membrane functionality leads to the loss of bacterial viability when exposed to the proton scavengers glycine, sarcosine and DMG at alkaline pH.

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

    PubMed

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

    2012-01-01

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

  5. Prevalence of acute mountain sickness in the Swiss Alps.

    PubMed Central

    Maggiorini, M; Bühler, B; Walter, M; Oelz, O

    1990-01-01

    OBJECTIVE--To assess the prevalence of symptoms and signs of acute mountain sickness of the Swiss Alps. DESIGN--A study using an interview and clinical examination in a representative population of mountaineers. Positive symptoms and signs were assigned scores to quantify the severity of acute mountain sickness. SETTING--Four huts in the Swiss Alps at 2850 m, 3050 m, 3650 m, and 4559 m. SUBJECTS--466 Climbers, mostly recreational: 47 at 2850 m, 128 at 3050 m, 82 at 3650, and 209 at 4559 m. RESULTS--In all, 117 of the subjects were entirely free of symptoms and clinical signs of acute mountain sickness; 191 had one or two symptoms and signs; and 158 had more than two. Those with more than two symptoms and signs were defined as suffering from acute mountain sickness. At 4559 m 11 climbers presented with high altitude pulmonary oedema or cerebral oedema, or both. Men and women were equally affected. The prevalence of acute mountain sickness correlated with altitude: it was 9% at 2850 m, 13% at 3050 m, 34% at 3650 m, and 53% at 4559 m. The most frequent symptoms and signs were insomnia, headache, peripheral oedema, and scanty pulmonary rales. Severe headache, vomiting, dizziness, tachypnoea, and pronounced pulmonary rales were associated with other symptoms and signs and therefore characteristic of acute mountain sickness. CONCLUSION--Acute mountain sickness is not an uncommon disease at moderately high altitude--that is, above 2800 m. Severe headache, vomiting, dizziness, tachypnoea, and pronounced pulmonary rales indicate severe acute mountain sickness, and subjects who suffer these should immediately descend to lower altitudes. PMID:2282425

  6. Onset of major Pleistocene glaciations in the Alps

    NASA Astrophysics Data System (ADS)

    Muttoni, G.; Carcano, C.; Garzanti, E.; Ghielmi, M.; Piccin, A.; Pini, R.; Rogledi, S.; Sciunnach, D.

    2003-04-01

    Since alligators patrolled Greenland swamps in the Eocene, the Earth's climate underwent significant cooling, which culminated in the Pleistocene Ice Age with recurring glaciations in vast regions of the Alps, Eurasia and North America, and overgrowth of polar icecaps in Antarctica and Greenland. During main Pleistocene glacial penetrations, the Alpine icecap invaded the low gradients of the Central Europe uplands and Italian Po plain. Peri-glacial sedimentary basins such as the Po Basin are natural collectors of past biological and climatic changes involving the waxing and waning of major icecaps. We have found in a 200m-thick core from the central Po plain near Milan stratigraphic evidence for a major glacial pulsation of the nearby Alpine icecap, which occurred in correspondence of a seismically traceable unconformity of regional relevance, termed the "Red Unconformity" (RU) in Eni/Agip terminology. The RU is associated with a major reorganization of vegetation cover and Alpine drainage pattern. The age of the RU was constrained magnetostratigraphically to the the first major Pleistocene glacio-eustatic low-stand at 0.87Ma (Oxygen Isotope Stage 22). This corresponds to the end of the "Mid Pleistocene Revolution" (MPR), a marked reorganization of northern hemisphere glaciation pattern which took place in the late Early Pleistocene. We suggest that the MPR/MIS 22 was associated with the onset of the first major Pleistocene glaciation in the Alps. Noticing the similarity in number of major Pleistocene glacieustatic low-stands starting with MIS 22, and the four-fold Alpine glacial subdivision of Penck and Brückner (1909), we conclude that "Penck and Brückner in 1909 may not have been, after all, that wrong" (Kukla and Cilek, 1996).

  7. Excessive lead burden among golden eagles in the Swiss Alps

    NASA Astrophysics Data System (ADS)

    Madry, Milena M.; Kraemer, Thomas; Kupper, Jacqueline; Naegeli, Hanspeter; Jenny, Hannes; Jenni, Lukas; Jenny, David

    2015-03-01

    Fragments from lead ammunition pose a poisoning risk for predators like golden eagles that scavenge on non-retrieved carcasses or offal left behind by hunters. Three golden eagles were found in the Swiss Alps with an acute lead poisoning. To investigate whether the few cases of lead-poisoned golden eagles are exceptional events or whether a substantial proportion of the Alpine golden eagle population is affected by lead at sublethal levels, we measured body burdens in golden eagles from Switzerland in comparison to eagle owls from the same area and to their respective prey. These two raptor species differ in their food as eagle owls feed on live-caught prey. Lead levels in soft tissues were significantly higher in golden eagles (median 1.14 μg g-1 dry weight in liver, 0.99 μg g-1 in kidney) than in eagle owls (0.14 and 0.23 μg g-1). Bones of golden eagles contained 10 times more lead (median of 12.45 μg g-1 dry weight) than owl bones (1.28 μg g-1), which represent substantially higher levels than previously reported for golden eagles. Bones of prey of both golden eagles and eagle owls had low lead concentrations. In order to investigate whether the sublethal lead of golden eagles originates from ammunition or from generic environmental contamination, we examined lead isotope ratios. Lead isotope signatures of golden eagle bones were very similar to those of ammunition, but differed from the signatures of bones of their prey, eagle owls and soil. Isotope signatures did not change with increasing bone lead concentration in golden eagles or any other group examined. These findings indicate that in the Alps, most golden eagles take up lead from spent ammunition in carcasses or their offal in sublethal quantities throughout their life and a few in lethal quantities leading to acute lead poisoning.

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

    PubMed

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

    2013-10-01

    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.

  9. FloodAlp! Frequency and climatic forcing of Holocene floods in the Central Alps - a project summary

    NASA Astrophysics Data System (ADS)

    Wirth, Stefanie B.; Glur, Lukas; Gilli, Adrian; Anselmetti, Flavio S.

    2014-05-01

    For a better assessment of the future Alpine flood hazard in the face of climate warming, an improvement of our understanding of the occurrence of floods in the past is important. The goal of the FloodAlp! project is thus to reconstruct the Holocene flood history of the Central Alps and to investigate the controlling climatic forcing, in particular regarding the flood occurrence during warmer and cooler climatic periods. 15 lake-sediment records served as flood archives, providing an Alpine transect from northeastern Switzerland to northern Italy covering a wide range in altitude (197 to 2065 m asl). The established flood catalog comprises over 4500 flood deposits, which are composed of terrestrial material mobilized by high fluvial runoff in the catchment areas. As the most important result, we found that during the past 10 kyrs the occurrence of Alpine floods was enhanced during cool climatic periods. Evidence is given by comparing our paleoflood record to independent reconstructions of air temperature and solar activity. This relation is explained by the latitudinal position of Atlantic and Mediterranean storm tracks, which generally shift southward during periods of low temperature/solar activity. In addition, we observe periods of enhanced flood occurrence in the Southern compared to the Northern Alps (most distinct from 4.2 to 2.4 kyr BP and during the Little Ice Age), suggesting a pronounced southward position of the storm tracks and/or blocking over the northern North Atlantic. Since such a setting resembles a negative NAO state, we interpret the South-Alpine flood frequency as a qualitative record of variations in a paleo-NAO pattern during the Holocene. This paleoclimatic evidence of a decreased Alpine flood frequency during warmer periods is most likely also applicable to warmer climatic conditions in the future. However, this result does not consider the intensity of the catalogued events. Studies addressing this issue are currently undertaken. Our

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

    PubMed

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

    2016-09-01

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

  11. The possible reasons for the absence of condensed phosphates in nature

    NASA Astrophysics Data System (ADS)

    Byrappa, K.

    1983-11-01

    Experiments to study the possible reasons for the absence of condensed phosphates in nature show that under hydrothermal conditions at P H2O>6 atm, the crystallization of condensed phosphates ceases and instead only simple orthophosphates crystallize irrespective of the temperature of the system. Similarly, even an increase in the concentration of alkali and alkaline earth elements in the system leads to decondensation of phosphates. It is well known that magmatic or postmagmatic processes of mineral formation are always accompanied by a certain amount of H2O and occur in the presence of many types of metals. This leads to the formation of only orthophosphates and not to condensed phosphates in nature.

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

    PubMed Central

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

    2016-01-01

    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 very slowly and pseudoreversion of the slow growth phenotype is common. 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. Our 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. 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. PMID:27965651

  13. Process for extracting technetium from alkaline solutions

    DOEpatents

    Moyer, Bruce A.; Sachleben, Richard A.; Bonnesen, Peter V.

    1995-01-01

    A process for extracting technetium values from an aqueous alkaline solution containing at least one alkali metal hydroxide and at least one alkali metal nitrate, the at least one alkali metal nitrate having a concentration of from about 0.1 to 6 molar. The solution is contacted with a solvent consisting of a crown ether in a diluent for a period of time sufficient to selectively extract the technetium values from the aqueous alkaline solution. The solvent containing the technetium values is separated from the aqueous alkaline solution and the technetium values are stripped from the solvent.

  14. Alkaline sorbent injection for mercury control

    DOEpatents

    Madden, Deborah A.; Holmes, Michael J.

    2002-01-01

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

  15. Alkaline sorbent injection for mercury control

    DOEpatents

    Madden, Deborah A.; Holmes, Michael J.

    2003-01-01

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

  16. Generally recognized as safe (GRAS) Lactococcus lactis strains associated with Lippia sidoides Cham. are able to solubilize/mineralize phosphate.

    PubMed

    de Lacerda, Jackeline Rossetti Mateus; da Silva, Thais Freitas; Vollú, Renata Estebanez; Marques, Joana Montezano; Seldin, Lucy

    2016-01-01

    Eight strains isolated from the stems of Lippia sidoides were identified as belonging to Lactococcus lactis, a bacterial species considered as "generally recognized as safe". Their capacity to solubilize/mineralize phosphate was tested in vitro with different inorganic and organic phosphorus (P) sources. All strains were able to solubilize calcium phosphate as an inorganic P source, and the best result was observed with strain 003.41 which solubilized 31 % of this P source. Rock phosphate, a mined rock containing high amounts of phosphate bearing minerals, was solubilized by five strains. When calcium phytate was the organic P source used, the majority of the strains tested showed phosphate mineralization activity. Moreover, all strains were able to solubilize/mineralize phosphate from poultry litter, a complex P source containing inorganic and predominantly organic P. The presence of genes coding for phytase and alkaline phosphatase was searched within the strains studied. However, only gene sequences related to alkaline phosphatase (phoA and phoD) could be detected in the majority of the strains (excepting strain 006.29) with identities varying from 67 to 88 %. These results demonstrate for the first time the potential of L. lactis strains for phosphate solubilization/mineralization activity using a broad spectrum of P sources; therefore, they are of great importance for the future development of more safe bioinoculants with possible beneficial effects for agriculture.

  17. AlpArray Austria - Illuminating the subsurface of Austria and understanding of Alpine geodynamics

    NASA Astrophysics Data System (ADS)

    Fuchs, Florian; Bokelmann, Götz; Bianchi, Irene; Apoloner, Maria-Theresia; AlpArray Working Group

    2015-04-01

    AlpArray Austria is a research project to study the geodynamics of the Eastern Alps and the subsurface of Austria with a large-scale mobile broadband seismological network. AlpArray Austria is part of the AlpArray project - a unique European transnational research initiative in which 64 research institutes from 17 countries join their expertise to advance our knowledge about the structure and evolution of the lithosphere beneath the entire Alpine area. AlpArray Austria is coordinated by the Department of Meteorology and Geophysics (IMGW) at the University of Vienna and funded by the Austrian Science Fund (FWF). During spring 2015 the deployment of 42 mobile broadband seismometers (and two permanent stations) will commence and by autumn 2015 Austria will be completely covered by a dense seismological network with an average station spacing of about 40 kilometers. The mobile network consisting of 27 Reftek 151 60s sensors (provided by IMGW) and 15 Trillium 120s instruments (provided by the Dublin Institute for Advanced Studies, DIAS, Ireland) will complement the permanent Austrian seismic network operated by the Austrian Zentralanstalt für Meteorologie und Geodynamik (ZAMG) that will also install the two permanent stations. AlpArray Austria will be continuously recording high-quality ground motion data for three years. Seismic data will be distributed through the European Integrated Data Archive (EIDA) and shared with the collaborating AlpArray institutes. AlpArray Austria will shed light on the detailed geological structure and geodynamical evolution of the Eastern Alps and the subsurface of Austria. Utilizing seismic analysis methods such as shear wave splitting, receiver functions and body wave dispersion the AlpArray Austria working group at IMGW will, together with the international partners, focus on seismic anisotropy in the upper mantle, the location of interfaces and tomography, to answer outstanding questions on slab geometry and subduction polarity under

  18. Role of bone-type tissue-nonspecific alkaline phosphatase and PHOSPO1 in vascular calcification.

    PubMed

    Bobryshev, Yuri V; Orekhov, Alexander N; Sobenin, Igor; Chistiakov, Dimitry A

    2014-01-01

    Matrix vesicle (MV)-mediated mineralization is important for bone ossification. However, under certain circumstances such as atherosclerosis, mineralization may occur in the arterial wall. Bone-type tissue-nonspecific alkaline phosphatase (TNAP) hydrolyzes inorganic pyrophosphate (PPi) and generates inorganic phosphate (Pi), which is essential for MV-mediated hydroxyapatite formation. MVs contain another phosphatase, PHOSPHO1, that serves as an additional supplier of Pi. Activation of bone-type tissue-nonspecific alkaline phosphatase (TNAP) in vascular smooth muscle cells precedes vascular calcification. By degrading PPi, TNAP plays a procalcific role changing the Pi/PPi ratio toward mineralization. A pathologic role of bone-type TNAP and PHOSPHO1 make them to be attractive targets for cardiovascular therapy.

  19. The significance of inhibitor-resistant alkaline phosphatase in the cytochemical demonstration of transport adenosine triphosphatase.

    PubMed

    Firth, J A; Marland, B Y

    1975-08-01

    The hydrolysis of disodium p-nitrophenyl phosphate at pH 9.0 by slices of formaldehydee-fixed rat renal cortex was investigated by colorimetric estimation of the nitrophenol liberated. It was found that three types of activity could be identified on the basis of their responses to inhibitors and cations: (a) alkaline phosphatase sensitive to inhibition by L-tetramisole; (b) potassium-dependent phosphatase, probably identifiable with the phosphatase component of sodium-potassium-dependent transport adenosine triphosphatase (?Na-K-ATPase); and (c) alkaline phosphatase insensitive to L-tetramisole. It was found that in the presence of strontium ions, as used in Na-K-ATPase cytochemistry, the activities of the second and third types of enzyme were approximately equal. The implications of these findings for the cytochemical demonstration of Na-K-ATPase are discussed.

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

    PubMed

    Okpere, E; Okorodudu, A; Gbinigie, O

    1988-01-01

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

  1. Human embryonic stem cells and macroporous calcium phosphate construct for bone regeneration in cranial defects in rats

    PubMed Central

    Liu, Xian; Wang, Ping; Chen, Wenchuan; Weir, Michael D.; Bao, Chunyun; Xu, Hockin H. K.

    2015-01-01

    Human embryonic stem cells (hESCs) are an exciting cell source to offer an unlimited supply of cells that can differentiate into all cell types for regenerative medicine applications. To date, there has been no report on hESCs with calcium phosphate cement (CPC) scaffolds for bone regeneration in vivo. The objectives of this study were to: (1) investigate hESCs for bone regeneration in vivo in critical-sized cranial defects in rats; (2) determine the effects of cell seeding and platelets in macroporous CPC on new bone and blood vessel formation. hESCs were cultured to yield mesenchymal stem cells (MSCs), which underwent osteogenic differentiation. Four groups were tested in rats: (1) CPC control without cells; (2) CPC with hESC-derived MSCs (“CPC+hESC-MSC”); (3) CPC with hESC-MSCs and 30% human platelet concentrate (hPC) (“CPC+hESC-MSC+30%hPC”); (4) CPC+hESC-MSC+50%hPC. In vitro, MSCs were derived from embryoid bodies (EBs) of hESCs. Cells on CPC were differentiated into the osteogenic lineage, with highly-elevated alkaline phosphatase (ALP) and osteocalcin (OC) expressions as well as mineralization. In vivo at 12 weeks, groups with hESC-MSCs and hPC had new bone 3-times, and blood vessel density 2-times, those of CPC control. The new bone in the defects contained osteocytes and blood vessels, and the new bone front was lined with osteoblasts. The group with 30% hPC and hESC-MSCs had a blood vessel density that was 49% greater than the hESC-MSC group without hPC, likely due to the various growth factors in the platelets enhancing both new bone and blood vessel formation. In conclusion, hESCs are promising for bone tissue engineering, and hPC can enhance new bone and blood vessel formation. Macroporous CPC with hESC-MSCs and hPC may be useful for bone regeneration in craniofacial and orthopedic applications. PMID:24972090

  2. Changes in Proton Dynamics in Articular Cartilage Caused by Phosphate Salts and Fixation Solutions.

    PubMed

    Zheng, Shaokuan; Xia, Yang

    2010-01-01

    The objective was to study the effect of phosphate salts and fixation solutions on the proton dynamics in articular cartilage in vitro. Microscopic magnetic resonance imaging (μMRI) T(2) anisotropy and nuclear magnetic resonance (NMR) double quantum-filtered (DQF) spectroscopy were used to study the full-thickness articular cartilage from several canine humeral heads. The in-plane pixel size across the depth of the cartilage tissue was 13 μm. The acid phosphate salt was an effective exchange catalyst for proton exchange in the cartilage with an organized structure of collagen fibrils, while the alkaline phosphate salt was not. For cartilage tissue containing less organized collagen fibrils, both acid and alkaline phosphate salts have no significant effect on the T(2) value at low concentration but decrease the T(2) value at high concentration. The solutions of NaCl, KCl, CaCl(2), and D-PBS were found to have no significant effect on T(2) and DQF in cartilage. This study demonstrates the ability to modify the proton exchange in articular cartilage using the solutions of phosphate salts. The ability to modify the proton exchange in articular cartilage can be used to modulate the laminar appearance of articular cartilage in MRI.

  3. Alkaline tolerant dextranase from streptomyces anulatus

    DOEpatents

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

    2003-01-01

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

  4. Technetium recovery from high alkaline solution

    DOEpatents

    Nash, Charles A.

    2016-07-12

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

  5. Toxicity of alkalinity to Hyalella azteca

    USGS Publications Warehouse

    Lasier, P.J.; Winger, P.V.; Reinert, R.E.

    1997-01-01

    Toxicity testing and chemical analyses of sediment pore water have been suggested for use in sediment quality assessments and sediment toxicity identification evaluations. However, caution should be exercised in interpreting pore-water chemistry and toxicity due to inherent chemical characteristics and confounding relationships. High concentrations of alkalinity, which are typical of sediment pore waters from many regions, have been shown to be toxic to test animals. A series of tests were conducted to assess the significance of elevated alkalinity concentrations to Hyalella azteca, an amphipod commonly used for sediment and pore-water toxicity testing. Toxicity tests with 14-d old and 7-d old animals were conducted in serial dilutions of sodium bicarbonate (NaHCO3) solutions producing alkalinities ranging between 250 to 2000 mg/L as CaCO3. A sodium chloride (NaCl) toxicity test was also conducted to verify that toxicity was due to bicarbonate and not sodium. Alkalinity was toxic at concentrations frequently encountered in sediment pore water. There was also a significant difference in the toxicity of alkalinity between 14-d old and 7-d old animals. The average 96-h LC50 for alkalinity was 1212 mg/L (as CaCO3) for 14-d old animals and 662 mg/L for the younger animals. Sodium was not toxic at levels present in the NaHCO3 toxicity tests. Alkalinity should be routinely measured in pore-water toxicity tests, and interpretation of toxicity should consider alkalinity concentration and test-organism tolerance.

  6. Mineral dust radiative effect on snow in European Alps

    NASA Astrophysics Data System (ADS)

    Di Mauro, Biagio; Fava, Francesco; Ferrero, Luca; Garzonio, Roberto; Baccolo, Giovanni; Delmonte, Barbara; Colombo, Roberto

    2015-04-01

    Mineral Dust (MD) is known to increase the absorption of solar radiation when deposited on snow and ice. This process causes a decrease in the albedo and may enhance snow melting, resulting in a positive radiative forcing (RF) in climate system. The RF from MD on snow can assume high values (~100-200 W/m2) after depositional events altering snow and ice radiative balance and hydrological cycle. In this study, we analyzed a significant MD transport happened during spring in 2014 and in particular its impact on snow optical properties. The dust plume was entrained in the troposphere over the Saharan desert (North African Grand Erg Oriental) during the passage of a cold front, and then transported NE over the Mediterranean by cyclonic atmospheric conditions. MD reached the European Alps where it was deposited by snowfall. We conducted a field proximal sensing survey in 10 plots (2x2 meters) at the Artavaggio plains (Lecco, Italy) with a hyperspectral radiometer (ASD Field-spec pro) collecting reflected radiance of snow in a spectral range between 350 and 2500 nm. Surface snow samples were collected and analyzed in clean room with microparticle counter in order to determine the size distribution and the concentration of MD in each sample. In addition, total mass of insoluble material was also measured by filtering the melted snow. Observed spectra were compared to those simulated by parameterizing the Snow, Ice, and Aerosol Radiation (SNICAR) radiative transfer model with observed variables such as snow grain size, snow density and size distribution of MD. We defined a novel spectral index, the Snow Darkening Index (SDI) to combine red and green wavelengths showing nonlinear correlation with measured MD concentration. Instantaneous radiative forcing was then estimated as the spectral difference between upwelling irradiance of plot containing MD and pure snow plots. MD concentration was up to 107 ppm and total mass of insoluble material up to 325 ppm. Measured RF values

  7. Alkaline Water and Longevity: A Murine Study

    PubMed Central

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

    2016-01-01

    The biological effect of alkaline water consumption is object of controversy. The present paper presents a 3-year survival study on a population of 150 mice, and the data were analyzed with accelerated failure time (AFT) model. Starting from the second year of life, nonparametric survival plots suggest that mice watered with alkaline water showed a better survival than control mice. Interestingly, statistical analysis revealed that alkaline water provides higher longevity in terms of “deceleration aging factor” as it increases the survival functions when compared with control group; namely, animals belonging to the population treated with alkaline water resulted in a longer lifespan. Histological examination of mice kidneys, intestine, heart, liver, and brain revealed that no significant differences emerged among the three groups indicating that no specific pathology resulted correlated with the consumption of alkaline water. These results provide an informative and quantitative summary of survival data as a function of watering with alkaline water of long-lived mouse models. PMID:27340414

  8. Performed surfactant-optimized aqueous alkaline flood

    SciTech Connect

    Thigpen, D.R.; Lawson, J.B.; Nelson, R.C.

    1991-11-26

    This paper describes improvement in a process for recovering oil from an acidic oil reservoir by injecting an aqueous alkaline solution comprising water, sodium chloride, and alkaline material for reacting with the reservoir oil forming a petroleum acid soap to form an in-situ surfactant system. The improvement comprises: selecting a preformed cosurfactant which is soluble in both the aqueous solution and the reservoir oil and has a solubility ratio which is grater than the solubility ratio of the petroleum acid soap where the solubility ratio is the ratio of solubility in the aqueous alkaline solution to the solubility in the reservoir oil; combining with the alkaline solution an amount of the preformed cosurfactant which will result in the in-situ surfacant system having a salinity about equal to a salinity which results in minimal interfacial tension between the oil in the reservoir and the in-situ surfactant system at reservoir temperature, wherein the amount of the preformed cosurfactant is about 0.3 percent by weight in the aqueous alkaline solution; and injecting the cosurfactant-aqueous alkaline solution mixture into the reservoir to displace oil toward a fluid production location.

  9. The 8.2 ka event in the northern Alps

    NASA Astrophysics Data System (ADS)

    Luetscher, M.; Hoffmann, D. L.; Müller, W.; Spötl, C.

    2012-04-01

    The 8.2 ka event has been identified as a widespread climate excursion affecting most of the Northern Hemisphere. High-resolution records from ice cores and speleothems constrain the chronology of this event to between 8.21±0.02 and 8.08±0.03 ka BP (Vinther et al. 2006, Cheng et al. 2009). A distinctive asymmetrical pattern in d18O is consistent with modelling results suggesting rapid input of freshwater into the northern Atlantic due to catastrophic drainage of ice-marginal lakes (LeGrande et al., 2008). Despite an increasing amount of data, the regional expression of this event is still poorly understood. Here, we present a new speleothem record from Gasselhöhle in the Northern Calcareous Alps, Austria. The 205 mm-long GAS19 stalagmite was analysed at high resolution for stable isotopes (100 µm) and trace elements (~10-15 µm; continuous LA-ICPMS profiles). Twelve individual MC-ICP-MS U/Th ages underline an annual growth rate of ca. 60 µm during the Early Holocene. The d18O averages -8.9‰, only slightly more depleted than modern carbonate precipitates from the same cave chamber. The 8.2 ka event is marked in GAS19 by a ca. 1‰ excursion with a minimum value of -9.9‰. Largely invariant trace element concentrations (e.g. Mg, U, Sr, Ba) indicate essentially no changes in the local hydrological regime and therefore support the hypothesis of a temperature-dominated signal. The proximity to the lacustrine isotope record from Mondsee (eg. Lauterbach et al. 2011) opens new perspectives for the interpretation of the oxygen isotope signal using two archives at different elevations. Moreover, several coeval speleothem records are available across the Eastern Alps fostering a spatial comparison of the proxy signals associated with this event. Cheng, H. et al. (2009), Geology, 37, 1007-1010 Lauterbach, S. et al. (2011), JQS, 26, 253-267 LeGrande, A.N., Schmidt, G.A. (2008), Paleoceanography, 23, doi: 10.1029/2008PA001610 Vinther, B. et al. (2006), JGR, 111, D13103

  10. Orogen parallel britlle extension in the Inner Northwestern Alps.

    NASA Astrophysics Data System (ADS)

    Champagnac, J. D.; Sue, C.; Delacou, B.; Burkhard, M.

    2003-04-01

    Rising attention has been recently paid to the latest extensional structures in the Alps, which took place under increasingly brittle conditions (e.g. Bistacchi, et al. 2000, Sue and Tricart 2002). Some of these structures seems to be still active . This brittle extension mainly occurs in the core of the arc, in the internal zones and could be linked to the ongoing extension observed under the light of seismotectonics studies (Sue, et al. 1999). Here we analyze the brittle deformation in the internal zone of the Northwestren Alps, from the Simplon fault zone (to the Northeast) to the Vanoise area (to the South). Our analysis is based on systematic mapping of the fault patterns, using remote sensing analysis combined with field work. The fault pattern we studied crosscuts all the ductile compression-related structures and piles of nappes. Thus, this brittle deformation postdates the ductile deformations, and is associated to one of the latter tectonic event in the belt during the recent-alpine history (Neogene times). The determination of paleostress field(s), based on the inversion of a large database of fault/stria measurements constrain the behaviour of this fault pattern: The results indicate a major large scale extensional paleostress field, in the NE-SW direction. In the South Valais, this orogen-parallel extension could be linked to the normal/dextral Simplon-Rhône fault zone dynamics. Further to the South, the paleostress fields also locally indicate NE-SW extension, but the driving forces remains a matter of debate. The orogen-parallel extension observed in this study have to be linked to the orogen-radial extension observed by (Sue and Tricart 2002) in the Briançonnais area, further to the South. We must also determine the relationship between the large scale brittle extension and the seismotectonics radial-to-the-arc extension. The geographic and/or temporal variations of the (paleo)stress fields remain to be constrain to precise the recent geodynamics

  11. Dendroclimatic trend and glacial fluctuations in the Central Italian Alps

    NASA Astrophysics Data System (ADS)

    Pelfini, M.; Santilli, M.; D Agata, C.; Diolaiuti, G.; Smiraglia, C.

    2003-04-01

    In the Alpine environment, one of the main limiting factors for tree growth is the thermal conditions of the vegetative season. The conifers at high altitude, if not subject to others disturbs, such as geomorphological processes or biological interferences, undergo a development, from which the width of annual rings depends. Five chronologies few centuries long, obtained for the species Larix decidua Mill. and Pinus cembra L. from different valleys of the Central Italian Alps (Alpisella, Valfurva, Gavia and Solda) in proximity of timberline (2000-2550 m of altitude), were analysed and their climatic signal gained; this last one was then related to the recent glacial fluctuations. The chronologies are the averages of many dendrochronological indicized curves obtained from dominant trees with regular growth and extended from 13th-17th century up to the present. The time intervals of the chronologies are the following ones: Pinus cembra: 1752-1999 for Valfurva; 1607-1999 for Gavia; 1593-1999 for Val Solda. With regard to Larix decidua: 1252-1998 for Val Solda; 1784-2001 for Alpisella. The good correspondence between the various chronologies allows to consider them representative of the climatic regional signal. In order to evidence climatic evolution, linear trends based on running mean with period of 11 years have been constructed. Those curves have been compared between them and then overlapped and mediated in order to obtain a climatic signal of regional value that excludes eventual local anomalies. Finally, the growth variations in the chronologies have been compared to known alpine climatic variations and glacial fluctuations. In particular time-distance curves (curves of cumulated frontal variations) of some glaciers from the Ortles-Cevedale Group were utilized. The periods of tree rings growth rate reduction appear well correlated to glacial advancing phases of the Little Ice Age and of the following phases. In particular, growth rate reductions are observable

  12. Modeling Sustainable Bioenergy Feedstock Production in the Alps

    NASA Astrophysics Data System (ADS)

    Kraxner, Florian; Leduc, Sylvain; Kindermann, Georg; Fuss, Sabine; Pietsch, Stephan; Lakyda, Ivan; Serrano Leon, Hernan; Shchepashchenko, Dmitry; Shvidenko, Anatoly

    2016-04-01

    Sustainability of bioenergy is often indicated by the neutrality of emissions at the conversion site while the feedstock production site is assumed to be carbon neutral. Recent research shows that sustainability of bioenergy systems starts with feedstock management. Even if sustainable forest management is applied, different management types can impact ecosystem services substantially. This study examines different sustainable forest management systems together with an optimal planning of green-field bioenergy plants in the Alps. Two models - the biophysical global forest model (G4M) and a techno-economic engineering model for optimizing renewable energy systems (BeWhere) are implemented. G4M is applied in a forward looking manner in order to provide information on the forest under different management scenarios: (1) managing the forest for maximizing the carbon sequestration; or (2) managing the forest for maximizing the harvestable wood amount for bioenergy production. The results from the forest modelling are then picked up by the engineering model BeWhere, which optimizes the bioenergy production in terms of energy demand (power and heat demand by population) and supply (wood harvesting potentials), feedstock harvesting and transport costs, the location and capacity of the bioenergy plant as well as the energy distribution logistics with respect to heat and electricity (e.g. considering existing grids for electricity or district heating etc.). First results highlight the importance of considering ecosystem services under different scenarios and in a geographically explicit manner. While aiming at producing the same amount of bioenergy under both forest management scenarios, it turns out that in scenario (1) a substantially larger area (distributed across the Alps) will need to be used for producing (and harvesting) the necessary amount of feedstock than under scenario (2). This result clearly shows that scenario (2) has to be seen as an "intensification

  13. SERUM VALUES OF ALKALINE PHOSPHATASE AND LACTATE DEHYDROGENASE IN EWING'S SARCOMA

    PubMed Central

    Baptista, André Mathias; Zumárraga, Juan Pablo; dos Santos, Renan Pires Negrão; Haubert, Guilherme de Oliveira; de Camargo, Olavo Pires

    2016-01-01

    ABSTRACT Objective: To study the relationship between the serum levels of alkaline phosphatase (AP) and lactate dehydrogenase (LDH), and the percentage of tumor necrosis (TN) in patients with Ewing´s Sarcoma (ES). Methods: This is a case series with retrospective evaluation of patients with diagnosis of ES divided into 2 groups: Group 1, patients whose serum levels of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were obtained in the staging phase before preoperative chemotherapy (CT), and Group 2, patients whose values were measured after completion of the preoperative CT. The percentage of tumor necrosis (TN) of surgical specimens extracted in surgery was also evaluated. Results: Eighty four medical records from 1995 to 2015 were included. Both AP as LDH decreased in the patients studied, the pre CT value being higher than the post CT value. The average decrease of LHD was 272.95 U/L and AP was 10.17 U/L. The average tumor necrosis was 65.12 %. There was no statistical correlation between serums levels and the tumor necrosis percentage. Conclusion: The serum levels values of AP and LDH are not predictors for chemotherapy-induced necrosis in patients with ES. Level of Evidence IV, Case Series. PMID:28243173

  14. Alkaline phosphatases. Newer concepts in isoenzymes and clinical applications.

    PubMed

    Griffiths, J

    1989-12-01

    ALP is a profoundly ubiquitous enzyme that originates in many, if not all, mammalian tissues. The development of a simple ISEF technique has allowed reflection of at least 12 cellular components. Improvements in technique may add further zones of information. The aura of darkness surrounding ALP is being slowly dissipated.

  15. pH-dependent phosphatization of ZnO nanoparticles and its influence on subsequent lead sorption.

    PubMed

    Xu, Huacheng; Li, Lina; Lv, Hua; Liu, Xin; Jiang, Helong

    2016-01-01

    Phosphatization of ZnO nanoparticles (ZNPs) at various pHs and its influence on subsequent lead sorption were investigated. Results showed that, in presence of phosphate, both the chemical speciation and crystalline phase of ZNPs were pH dependent that most of them were converted to crystalline Zn3(PO4)2 at acidic pHs, but only little amorphous hopeites can be formed under alkaline condition. Phosphatization process significantly enhanced subsequent lead sorption with the order of acidic process > alkaline > pristine ZNPs. Spectroscopic analysis including ATR-FTIR and XPS revealed main mechanisms of lead phosphate precipitation and inner-sphere complexes for lead sorption on acidic and alkaline treatment products, respectively. The potential toxicity of ZNPs and heavy metals in eutrophic aquatic ecosystems would thus be reduced due to the ubiquitous phosphatization process. This study highlights the importance of environmental variables in exploring the environmental behavior and fate of heavy metals as well as nanoparticles in natural waters.

  16. Trypanosoma rangeli: an alkaline ecto-phosphatase activity is involved with survival and growth of the parasite.

    PubMed

    Dos-Santos, André L A; Dick, Claudia F; Silveira, Thaís S; Fonseca-de-Souza, André L; Meyer-Fernandes, José R

    2013-10-01

    The aim of this work was to investigate whether an alkaline ecto-phosphatase activity is present in the surface of Trypanosoma rangeli. Intact short epimastigote forms were assayed for ecto-phosphatase activity to study kinetics and modulators using β-glycerophosphate (β-GP) and p-nitrophenyl phosphate (pNPP) as substrates. Its role in parasite development and differentiation was also studied. Competition assays using different proportions of β-GP and pNPP evidenced the existence of independent and non-interacting alkaline and acid phosphatases. Hydrolysis of β-GP increased progressively with pH, whereas the opposite was evident using pNPP. The alkaline enzyme was inhibited by levamisole in a non-competitive fashion. The Ca(2+) present in the reaction medium was enough for full activity. Pretreatment with PI-PLC decreased the alkaline but not the acid phosphatase evidence that the former is catalyzed by a GPI-anchored enzyme, with potential intracellular signaling ability. β-GP supported the growth and differentiation of T. rangeli to the same extent as high orthophosphate (Pi). Levamisole at the IC50 spared significantly parasite growth when β-GP was the sole source of Pi and stopped it in the absence of β-GP, indicating that the alkaline enzyme can utilize phosphate monoesters present in serum. These results demonstrate the existence of an alkaline ecto-phosphatase in T. rangeli with selective requirements and sensitivity to inhibitors that participates in key metabolic processes in the parasite life cycle.

  17. Rock phosphate enriched compost: an approach to improve low-grade Indian rock phosphate.

    PubMed

    Biswas, D R; Narayanasamy, G

    2006-12-01

    In this study, rock phosphate enriched composts (RP-compost) were prepared by mixing four low-grade Indian rock phosphates with rice straw with and without Aspergillus awamori. RP-compost had higher total P, citrate soluble P (CSP), organic P (Org.P), acid and alkaline phosphatase activities, and lower water soluble P (WSP) and microbial biomass C (MBC) than normal compost. Inoculation with A. awamori increased total P, WSP, CSP, Org.P, MBC and acid phosphatase activity. RP-compost recorded lower Olsen P at the initial period of incubation study than diammonium phosphate (DAP), but improved significantly with the progress of time. RP-compost prepared at 4% charged rate resulted in higher Olsen P throughout the incubation period compared to 2% charged rate. Similar trend were obtained with those RP-composts prepared with A. awamori. Data on pot experiment revealed higher yield and P uptake by mungbean (Vigna radiata) due to addition of RP-composts over control. The effectiveness of RP-compost ranged from 61.4% (MussoorieRP-compost) to 94.1% (PuruliaRP-compost) as that of DAP on dry matter yield and 48.8% (JhabuaRP-compost) to 83.7% (PuruliaRP-compost) on total P uptake. Enriched compost prepared at 4% charged rate recorded 15.8% and 10.6% extra yield and P uptake, respectively by mungbean over 2% charged compost. Also RP-compost inoculated with A. awamori resulted in 13.0 and 21.5% extra yield and P uptake than without A. awamori treated group. Thus, RP enriched compost could be an alternative and viable technology to utilize both low-grade RPs and rice straw efficiently.

  18. 21 CFR 184.1301 - Ferric phosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Ferric phosphate. 184.1301 Section 184.1301 Food... GRAS § 184.1301 Ferric phosphate. (a) Ferric phosphate (ferric orthophosphate, iron (III) phosphate, Fe... from one to four molecules of water of hydration. It is prepared by reaction of sodium phosphate...

  19. Kinematics of the Alpenrhein-Bodensee graben system in the Central Alps: Oligocene/Miocene transtension due to formation of the Western Alps arc

    NASA Astrophysics Data System (ADS)

    Ring, Uwe; Gerdes, Axel

    2016-06-01

    We report fault slip data from exhumed fault surfaces along the NNE trending Alpenrhein valley and its intersection with the WNW striking Bodensee Graben near the Alpine thrust front of the Central Alps in the Swiss-Austrian-German border region. This conjugated graben system straddles the boundary between the Alps and its foreland and allows comparing the kinematics of graben formation between the two different tectonic domains. Our data show sinistral transtension along the Alpenrhein Graben and dextral transtension along the Bodensee Graben. Both transtensional graben systems resulted from the same kinematic regime of NW directed shortening and NE oriented extension. The graben faults are not older than NW striking tear faults associated with NW directed Oligocene nappe emplacement in the Helvetics starting at 35-30 Ma and ending by 25-20 Ma. Compatible with this are six U-Pb ages of calcite fibers from four samples yielding consistent ages ranging from 25.3 ± 5.6 Ma to 21.8 ± 3.4 Ma (2σ errors). Earthquake data since 1996 show that kinematic directions persisted until the Recent. Our data broadly fit with the kinematic evolution of the Oberrhein Graben, which shows significant E-W extension in the Oligocene. We suggest that Oligocene extension in the Alps and its foreland resulted from the increased curvature of the Western Alps arc and associated moderate tangential stretching in the internal parts of the Central Alps. We discuss a tectonic model of eastward rollback of the west Mediterranean subduction zone associated with counterclockwise rotation of Adria, the latter of which aided the formation of the Western Alps arc.

  20. 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.