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Sample records for alkaline phosphate alp

  1. ALP (Alkaline Phosphatase) Test

    MedlinePlus

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

  2. Alkaline-resistance model of subtilisin ALP I, a novel alkaline subtilisin.

    PubMed

    Maeda, H; Mizutani, O; Yamagata, Y; Ichishima, E; Nakajima, T

    2001-05-01

    The alkaline-resistance mechanism of the alkaline-stable enzymes is not yet known. To clarify the mechanism of alkaline-resistance of alkaline subtilisin, structural changes of two typical subtilisins, subtilisin ALP I (ALP I) and subtilisin Sendai (Sendai), were studied by means of physicochemical methods. Subtilisin NAT (NAT), which exhibits no alkaline resistance, was examined as a control. ALP I gradually lost its activity, accompanied by protein degradation, but, on the contrary, Sendai was stable under alkaline conditions. CD spectral measurements at neutral and alkaline pH indicated no apparent differences between ALP I and Sendai. A significant difference was observed on measurement of fluorescence emission spectra of the tryptophan residues of ALP I that were exposed on the enzyme surface. The fluorescence intensity of ALP I was greatly reduced under alkaline conditions; moreover, the reduction was reversed when alkaline-treated ALP I was neutralized. The fluorescence spectrum of Sendai remained unchanged. The enzymatic and optical activities of NAT were lost at high pH, indicating a lack of functional and structural stability in an alkaline environment. Judging from these results, the alkaline resistance is closely related to the surface structure of the enzyme molecule.

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

    PubMed Central

    Jazaeri, Mina; Malekzadeh, Hosein; Abdolsamadi, Hamidreza; Rezaei-Soufi, Loghman; Samami, Mohammad

    2015-01-01

    Although salivary alkaline phosphatase (ALP) can balance deand remineralization processes of enamel, there is no evidence regarding its effects on the concentrations of calcium and phosphate in saliva. The present study aims to determine the relationship between salivary ALP activity and the concentrations of calcium and phosphate in saliva. In this cross-sectional study, we evaluated salivary markers in 120 males, ages 19 to 44 years. All participants provided 5 mL of unstimulated whole saliva and the level of enzyme activity as well as calcium and phosphate concentrations were measured using a colorimetric method. Data were gathered and analyzed by statistical package for social sciences (SPSS) 13.00 using Pearson correlation test. A p value of <0.05 was considered statistically significant. The mean age of participants in the present study was 32.95 ± 8.09 years. The mean pH of saliva was 6.65 ± 0.62. Salivary parameters included average ALP activity (5.04 ± 1.866 U/dL), calcium (4.77 ± 0.877 mg/dL) and phosphate (10.38 ± 2.301 mg/dL). Pearson correlation test showed no significant relationship between ALP activity and calcium and phosphate concentrations in saliva (p>0.05). According to the results of the present study, there was no significant relation between salivary ALP activity and calcium and phosphate concentrations in saliva. However, further research is highly recommended. PMID:25870846

  4. Alkaline phosphatase assay using a near-infrared fluorescent substrate merocyanine 700 phosphate.

    PubMed

    Gong, Haibiao; Little, Garrick; Cradduck, Mark; Draney, Daniel R; Padhye, Nisha; Olive, D Michael

    2011-05-15

    Alkaline phosphatase (ALP) is a phosphomonoester hydrolase that is commonly used as a conjugating enzyme in biological research. A wide variety of substrates have been developed to assay its activity. In this study, we developed an ALP assay method utilizing merocyanine 700 (MC700) based substrate MC700 phosphate (MC700p). MC700 is a near-infrared fluorescent merocyanine dye, and has excitation/emission maxima at 686 nm/722 nm in ALP assay buffer. Upon hydrolysis by ALP, MC700p is converted to MC700. The fluorescence of MC700 is dependent on the pH and detergent concentration in the buffer. The fluorescence signal produced by MC700p hydrolysis is linearly related to the ALP amount and substrate concentration. A stop solution containing EDTA could be used to stop the ALP/MC700p reaction. It was also demonstrated that MC700p could substitute pNpp as the ALP substrate in a commercial 17β-Estradiol enzyme immunoassay kit. PMID:21482307

  5. Alkaline phosphatase (tissue-nonspecific isoenzyme) is a phosphoethanolamine and pyridoxal-5'-phosphate ectophosphatase: normal and hypophosphatasia fibroblast study.

    PubMed Central

    Fedde, K N; Whyte, M P

    1990-01-01

    To clarify its physiologic role, alkaline phosphatase (ALP) was examined in normal skin fibroblasts and was shown to be the tissue-nonspecific (TNS) isoenzyme type (as evidenced by heat and inhibition profiles) and to be active toward millimolar concentrations of the putative natural substrates phosphoethanolamine (PEA) and pyridoxal-5'-phosphate (PLP). Fibroblast ALP has a low-affinity activity, with a distinctly alkaline pH optimum (9.3), toward 4-methylumbelliferyl phosphate (4-MUP), PEA, and PLP but a more physiologic pH optimum (8.3) toward physiologic concentrations (micromolar) of PEA and PLP. Normal fibroblast ALP is linked to the outside of the plasma membrane, since in intact cell monolayers (1) dephosphorylation rates of the membrane-impermeable substrates PEA and PLP in the medium at physiologic pH were similar to those observed with disrupted cell monolayers, (2) brief exposure to acidic medium resulted in greater than 90% inactivation of the total ALP activity, and (3) digestion with phosphatidylinositol-specific phospholipase C (PI-PLC) released about 80% of the ALP activity. Hypophosphatasia fibroblasts were markedly deficient (2%-5% control values) in alkaline and physiologic ALP activity when 4-MUP, PLP, and PEA were used as substrate. The majority of the detectable ALP activity, however, appeared to be properly lipid anchored in ecto-orientation. Thus, our findings of genetic deficiency of PEA- and PLP-phosphatase activity in hypophosphatasia fibroblasts, as well as our biochemical findings, indicate that TNS-ALP acts physiologically as a lipid-anchored PEA and PLP ectophosphatase. PMID:2220817

  6. 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...

  7. 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...

  8. 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...

  9. Alkaline and alkaline earth metal phosphate halides and phosphors

    SciTech Connect

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

    2012-11-13

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

  10. Mineralization of alkaline phosphatase-complexed collagen implants in the rat in relation to serum inorganic phosphate.

    PubMed

    van den Bos, T; Oosting, J; Everts, V; Beertsen, W

    1995-04-01

    The present study was designed to determine the relationship between mineralization of collagenous matrices and serum levels of calcium and inorganic phosphate. Collagen slices were prepared from bovine dentin or cortical bone and complexed with varying amounts of intestinal alkaline phosphatase (ALP). The enzyme was added to induce de novo mineralization. The ALP-complexed slices were implanted subcutaneously over the skull and in the dorsolateral aspect of the abdominal wall in female Wistar rats of various ages (5-, 10-, 20-, or 35-week-old) and in young male rats fed on a low-P diet. After 1-4 weeks, the implants were removed and analyzed for calcium and phosphate content. In addition, serum levels of calcium and phosphate (total and inorganic) were determined. It was shown that the highest mineral influx occurred in the younger rats (which were also highest in serum P(i)), whereas almost no mineral uptake occurred in the older ones. Also in rats fed on a low-P diet (which were low in serum P(i), a strongly decreased mineral influx was noted. In all animal groups a positive correlation was found between the degree of mineralization and serum P(i). No distinct relationship was found between serum Ca/organic phosphate levels and mineral influx in the implants. In vitro incubation of ALP-collagen conjugates in serum from younger and older rats confirmed our view that serum P(i), besides local levels of ALP, is important in de novo mineral deposition. For accretion of mineral in partially remineralized collagenous carriers, ALP activity was not required.

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

    PubMed

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

    2016-10-01

    In this paper, a novel colorimetric method for the detection of alkaline phosphatase (ALP) and pyrophosphate (PPi) was designed based on a Cu(2+)-horseradish peroxidase (HRP)-3,3',5,5'-tetra-methylbenzidine (TMB)-H2O2 system. In the presence of ALP, l-ascorbic acid-2-phosphate (AAP) could be hydrolyzed to ascorbic acid which could reduce Cu(2+) to Cu(+) to inhibit the enzymatic activity of HRP in the colorimetric system. The change in absorbance was found to be proportional to the ALP concentration with a linear detection range and a limit of detection of 5.4 mU mL(-1). In the presence of PPi, because Cu(2+) was chelated by PPi, the conversion of Cu(ii) by AA was effectively inhibited. The color of the HRP-TMB-H2O2 system with Cu(2+) showed blue. The HRP-TMB-H2O2 system with the Cu(2+) colorimetric system could also detect PPi with a satisfying result. In summary, this method possesses sensitivity, reproducibility, and cost-effectiveness without labelling and separation and the use of a colorimetric method is more in line with the requirements of on-site detection and green chemistry. PMID:27412643

  12. Lactate dehydrogenase (LD), alkaline phosphatase (ALP) isoenzymatic patterns in Iraqi children with visceral leishmaniasis before and after treatment with stibogluconate.

    PubMed

    Taher, Jasim Hameed; Al-Mulla Hummadi, Yassir Mustafa Kamal; Al-Bashir, Nada Muhammed Taha; Al-Araji, Ali Shaalan

    2016-06-01

    The mean levels of alkaline phosphatase (ALP), lactate dehydrogenase enzymes exhibited a significant elevation in visceral leishmaniasis (VL) patients compared to the control. There was no significant change in relation to the sex and age. ALP isoenzymes revealed three banding patterns which differ from the three zymodems which were obtained from control group. These differences may be due to isoenzymes activity of patients with VL before and after therapy. Lactate dehydrogenase (LD) isoenzymes revealed five banding patterns differ from the five normal zymodems. These differences mainly occurred due to LD isoenzymes activity in patients with VL before and after therapy. PMID:27413293

  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. Cortisol modification of HeLa 65 alkaline phosphatase. Decreased phosphate content of the induced enzyme.

    PubMed

    Bazzell, K L; Price, G; Tu, S; Griffin, M

    1976-01-15

    Alkaline phosphatase activity of HeLa cells is increased 5-20-fold during growth in medium with cortisol. The increase in enzyme activity is due to an enhanced catalytic efficiency rather than an increase in alkaline phosphatase protein in induced cells. In the present study the chemical composition of control and induced forms of alkaline phosphatase were investigated to determine the enzyme modification that may be responsible for the increased catalytic activity. HeLa alkaline phosphatase is a phosphoprotein and the induced form of the enzyme has approximately one-half of the phosphate residues associated with control enzyme. The decrease in phosphate residues of the enzyme apparently alters its catalytic activity. Other chemical components of purified alkaline phosphatase from control and induced cells are similar; these include sialic acid, hexosamine and sulfhydryl residues. PMID:1248469

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

    PubMed

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

    1995-08-01

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

  16. 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.

  17. Phosphate glass electrode with good selectivity for alkaline-earth cations

    USGS Publications Warehouse

    Truesdell, A.H.; Pommer, A.M.

    1963-01-01

    A phosphate glass has been found to have a significant electrode specificity toward alkaline-earth ions. The order of selectivity is 2H + > Ba++ > Sr++ > Ca++ > 2K+ > 2Na+ > Mg++. Exchange properties are discussed in relation to possible structure. Its use to determine activity of Ca++ in natural systems containing Mg++ is suggested.

  18. ALP - blood test

    MedlinePlus

    ... features on this page, please enable JavaScript. Alkaline phosphatase (ALP) is a protein found in all body ... II Pancreatitis Renal cell carcinoma Alternative Names Alkaline phosphatase References Berk P, Korenblat K. Approach to the ...

  19. 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.

  20. Spectroscopic studies of copper doped alkaline earth lead zinc phosphate glasses

    NASA Astrophysics Data System (ADS)

    Sastry, S. Sreehari; Rao, B. Rupa Venkateswara

    2014-02-01

    In this paper spectroscopic investigation of Cu2+ doped alkaline earth lead zinc phosphate glasses was done through the spectroscopic techniques like X-ray diffraction, Ultra Violet (UV) absorption Spectroscopy, Electron Paramagnetic Resonance (EPR - X band), Fourier Transform Infra Red (FTIR) and Raman Spectroscopy. Alkaline earth lead zinc phosphate glasses containing 0.1% copper oxide (CuO) were prepared by the melt quenching technique. Spectroscopic studies indicated that there is a greater possibility for the copper ions to exist in Cu2+ state in these glasses. The optical absorption spectra indicated that the absorption peak of Cu2+ is a function of composition. The maxima absorption peak was reported at 862 nm for strontium lead zinc phosphate glass. Bonding parameters were calculated for the optical and EPR data. All these spectral results indicated clearly that there are certain structural changes in the present glass system with different alkaline earth contents. The IR and Raman spectra noticed the breaking of the P-O-P bonds and creating more number of new P-O-Cu bonds.

  1. 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.

  2. Cellular Performance Comparison of Biomimetic Calcium Phosphate Coating and Alkaline-Treated Titanium Surface

    PubMed Central

    Wei, Mei

    2013-01-01

    The influence of biomimetic calcium phosphate coating on osteoblasts behavior in vitro is not well established yet. In this study, we investigated the behavior of osteoblastic rat osteosarcoma 17/2.8 cells (ROS17/2.8) on two groups of biomaterial surfaces: alkaline-treated titanium surface (ATT) and biomimetic calcium phosphate coated ATT (CaP). The cell attachment, proliferation, differentiation, and morphology on these surfaces were extensively evaluated to reveal the impact of substrate surface on osteoblastic cell responses. It was found that the ROS17/2.8 cells cultured on the ATT surface had higher attachment and proliferation rates compared to those on the CaP surface. Our results also showed that the calcium phosphate coatings generated in this work have an inhibiting effect on osteoblast adhesion and further influenced the proliferation and differentiation of osteoblast compared to the ATT surface in vitro. Cells on the ATT surface also exhibited a higher alkaline phosphatase activity than on the CaP surface after two weeks of culture. Immunofluorescence staining and scanning electron microscopy results showed that the cells adhered and spread faster on the ATT surface than on the CaP surface. These results collectively suggested that substrate surface properties directly influence cell adhesion on different biomaterials, which would result in further influence on the cell proliferation and differentiation. PMID:24455730

  3. Responses of Phosphate Transporter Gene and Alkaline Phosphatase in Thalassiosira pseudonana to Phosphine

    PubMed Central

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

    2013-01-01

    Phosphine, which is released continuously from sediment, can affect the eco-physiological strategies and molecular responses of phytoplankton. To examine the effects of phosphine on phosphorus uptake and utilization in Thalassiosira pseudonana, we examined the transcriptional level of the phosphate transporter gene (TpPHO) and the activity of alkaline phosphatase (AKP) in relation to supplement of various concentrations of phosphine. TpPHO expression was markedly promoted by phosphine in both the phosphate-deficient and phosphate-4 µM culture. However, high phosphine concentrations can inhibit TpPHO transcription in the declining growth phase. AKP activity was also higher in the phosphine treatment groups than that of the control. It increased with increasing phosphine concentration in the range of 0 to 0.056 µM but was inhibited by higher levels of phosphine. These responses revealed that phosphine can affect phosphate uptake and utilization in T. pseudonana. This result was consistent with the effect of phosphine on algal growth, while TpPHO expression and AKP were even more sensitive to phosphine than algal growth. This work provides a basic understanding for further research about how phosphine affects phytoplankton. PMID:23544096

  4. Evaluation of yolk protein levels as estrogenic biomarker in bivalves; comparison of the alkali-labile phosphate method (ALP) and a species-specific immunoassay (ELISA).

    PubMed

    Morthorst, Jane E; Holbech, Henrik; Jeppesen, Morten; Kinnberg, Karin L; Pedersen, Knud L; Bjerregaard, Poul

    2014-11-01

    Altered concentration of the vertebrate yolk protein precursor vitellogenin is a recognized biomarker for endocrine disruption in fish, and within recent years yolk protein alteration has also been associated with endocrine disruption in bivalves. Species-specific, direct and sensitive methods for quantification of vitellogenin in fish have been available for years whereas bivalve yolk protein levels have been estimated indirectly by alkali-labile phosphate (ALP) liberated from high molecular weight proteins because the sequence and biochemical structure of most bivalve yolk proteins are unknown. By applying a species-specific enzyme-linked immunosorbent assay (ELISA) for accurate determination of yolk protein level the impact of 17β-estradiol (57, 164 and 512 ng/L) on the freshwater bivalve Unio tumidus was investigated and compared with ALP estimations. Seven weeks of exposure during the pre-spawning and spawning period had no consistent effect on yolk protein concentration in hemolymph, and ALP levels in hemolymph also remained unchanged in both males and females. Further, basal male and female ALP levels were indistinguishable whereas the ELISA demonstrated that yolk protein levels of females exceeded male levels at the time of sampling, although male basal levels were high compared to fish. Altogether it is shown that individual ALP levels do not reflect yolk protein levels and hence hemolymph ALP levels cannot serve as biomarker for estrogenic exposure during the pre-spawning and spawning period in U. tumidus. The necessity of sensitive and validated biomarkers for reliable interpretation of data and the utility of ALP and yolk protein levels as biomarkers in bivalves are discussed. PMID:25066673

  5. Direct determination of total mercury in phosphate rock using alkaline fusion digestion.

    PubMed

    D'Agostino, Fabio; Oliveri, Elvira; Bagnato, Emanuela; Falco, Francesca; Mazzola, Salvatore; Sprovieri, Mario

    2014-12-10

    The aim of this work was to develop a new method to determine the mercury (Hg) concentrations in phosphate rock using a dedicated analytical instrument (the DMA80 Tricell by Milestone) that employs an integrated sequence of thermal decomposition followed by catalyst conversion, amalgamation and atomic absorption spectrophotometry. However, this instrument underestimates Hg concentrations when phosphorite and apatite rocks are investigated with a classic thermal decomposition treatment that complies with US EPA method 7473. Therefore, to improve the recovery of total Hg, we performed alkaline fusion digestion (AFD) directly inside the furnace of the instrument, using BCR(32) as a certified reference material (Moroccan phosphate rock--phosphorite). The salts used for the AFD were a mixture of Na2CO3, K2CO3 and Li2CO3, which melt at about 400°C, due to their ability to form a ternary eutectic and to decompose the phosphorite matrices at 700°C. By adopting this analytical approach, the Hg recovery in BCR(32) was about 100%, compared to 40% when the reference material was analysed without using the alkaline fusion salt. We suggest that the AFD allowed the decomposition of the sample matrix and that some Hg compounds linked with other functional groups may be transformed in carbonates that sublimate at lower temperatures than other Hg compounds. This original method was tested on a number of different geological samples to compare the differences between the AFD method and the thermal treatment in order to verify the working range and to check the robustness of the new approach.

  6. XANES Demonstrates the Release of Calcium Phosphates from Alkaline Vertisols to Moderately Acidified Solution.

    PubMed

    Andersson, Karl O; Tighe, Matthew K; Guppy, Christopher N; Milham, Paul J; McLaren, Timothy I; Schefe, Cassandra R; Lombi, Enzo

    2016-04-19

    Calcium phosphate (CaP) minerals may comprise the main phosphorus (P) reserve in alkaline soils, with solubility dependent on pH and the concentration of Ca and/or P in solution. Combining several techniques in a novel way, we studied these phenomena by progressively depleting P from suspensions of two soils (low P) using an anion-exchange membrane (AEM) and from a third soil (high P) with AEM together with a cation-exchange membrane. Depletions commenced on untreated soil, then continued as pH was manipulated and maintained at three constant pH levels: the initial pH (pHi) and pH 6.5 and 5.5. Bulk P K-edge X-ray absorption near-edge structure (XANES) spectroscopy revealed that the main forms of inorganic P in each soil were apatite, a second more soluble CaP mineral, and smectite-sorbed P. With moderate depletion of P at pHi or pH 6.5, CaP minerals became more prominent in the spectra compared to sorbed species. The more soluble CaP minerals were depleted at pH 6.5, and all CaP minerals were exhausted at pH 5.5, showing that the CaP species present in these alkaline soils are soluble with decreases of pH in the range achievable by rhizosphere acidification. PMID:26974327

  7. Phase Stability of Chromium(III) Oxide Hydroxide in Alkaline Sodium Phosphate Solutions

    SciTech Connect

    S.E. Ziemniak; E.P. Opalka

    2003-07-08

    Grimaldiite ({alpha}-CrOOH) is shown to transform to a sodium-chromium(III)-hydroxyphosphate compound (SCHP) in alkaline sodium phosphate solutions at elevated temperatures via CrOOH(s) + 4Na{sup +} + 2HPO{sub 4}{sup 2-} = Na{sub 4}Cr(OH)(PO{sub 4}){sub 2}(s) + H{sub 2}O. X-ray diffraction analyses indicate that SCHP possesses an orthorhombic lattice having the same space group symmetry (Ibam, No.72) as sodium ferric hydroxyphosphate. A structurally-consistent designation for SCHP is Na{sub 3}Cr(PO{sub 4}){sub 2} {center_dot} NaOH; the molar volume of SCHP is estimated to be 1552 cm{sup 3}. The thermodynamic equilibrium for the above reaction was defined in the system Na{sub 2}O-P{sub 2}O{sub 5}-Cr{sub 2}O{sub 3}-H{sub 2}O for Na/P molar ratios between 2.0 and 2.4. On the basis of observed reaction threshold values for sodium phosphate concentration and temperature, the standard molar entropy (S{sup o}), heat capacity (C{sub p}{sup o}) and free energy of formation ({Delta}G{sub f}{sup o}) for SCHP were calculated to be 690 J/(mol-K), 622 J/(mol-K) and -3509.97 kJ/mol, respectively.

  8. 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.

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

    SciTech Connect

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

    1994-04-01

    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]- [l reversible] NaZnPO[sub 4](s) + H[sub 2]O or 2ZnO(s) + H[sub 3]PO[sub 4](aq) [l 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, 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. On the basis of 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 -1510.6 kJ/mol, respectively; similar values for Zn[sub 2](OH)PO[sub 4] (tarbuttite) were 235.9 J/(mol K) and -1604.6 kJ/mol. Additions of sodium sulfite and sulfate did not alter the above reactions. 26 refs., 5 figs., 7 tabs.

  10. Phosphate Sorption and Desorption on Pyrite in Primitive Aqueous Scenarios: Relevance of acidic → Alkaline Transitions

    NASA Astrophysics Data System (ADS)

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

    2007-02-01

    Phosphate (P i) sorption assays onto pyrite in media simulating primeval aquatic scenarios affected by hydrothermal emissions, reveal that acidic conditions favour P i sorption whereas mild alkaline media as well as those simulating sulfur oxidation to SO2- 4 revert this capture process. Several mechanisms relevant to P i availability in prebiotic eras are implicated in the modulation of these processes. Those favouring sorption are: (a) hydrophobic coating of molecules, such as acetate that could be formed in the vicinity of hydrothermal vents; (b) water and Mg2+ bridging in the interface mineral-aqueous media; (c) surface charge neutralization by monovalent cations (Na+ and K+). The increase of both the medium pH and the SO2- 4 trapping by the mineral interface would provoke the release of sorbed P i due to charge polarization. Moreover it is shown that P i self-modulates its sorption, a mechanism that depends on the abundance of SO2- 4 in the interface. The relevance of the proposed mechanisms of P i capture, release and trapping arises from the need of abundant presence of this molecule for primitive phosphorylations, since similarly to contemporary aqueous media inorganic phosphate concentrations in primitive seas should have been low. It is proposed that the presence of sulphide minerals with high affinity to P i could have trapped this molecule in an efficient manner, allowing its concentration in specific niches. In these niches, the conditions studied in the present work would have been relevant for its availability in soluble form, specially in primitive insulated systems with pH gradients across the wall.

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

    PubMed

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

    2007-02-01

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

  12. Occurrence of salt, pH, and temperature-tolerant, phosphate-solubilizing bacteria in alkaline soils

    PubMed

    Johri; Surange; Nautiyal

    1999-08-01

    An ecological survey was conducted to characterize 4800 bacterial strains isolated from the root-free soil, rhizosphere, and rhizoplane of Prosopis juliflora growing in alkaline soils. Of the 4800 bacteria, 857 strains were able to solubilize phosphate on plates. The incidence of phosphate-solubilizing bacteria (PSB) in the rhizoplane was highest, followed by rhizosphere and root-free soil. Eighteen bacterial strains out of 857 PSB were able to produce halo at 30 degrees C in a plate assay in the presence of 5% salt (NaCl) and solubilize tricalcium phosphate in National Botanical Research Institute's phosphate growth medium (NBRIP) broth, in the presence of various salts, pHs, and temperatures. Among the various bacteria tested, NBRI4 and NBRI7 did not produced halo in a plate assay at 30 degrees C in the absence of salt. Contrary to indirect measurement of phosphate solubilization by plate assay, the direct measurement of phosphate solubilization in NBRIP broth assay always resulted in reliable results. The phosphate solubilization ability of NBRI4 was higher than in the control in the presence of salts (NaCl, CaCl2, and KCl) at 30 degrees C. Phosphate solubilization further increased in the presence of salts at 37 degrees C as compared with 30 degrees C. At 37 degrees C, CaCl2 reduced phosphate solubilization ability of NBRI4 compared with the control. The results indicated the role of calcium salt in the phosphate solubilization ability of NBRI4.http://link.springer-ny. com/link/service/journals/00284/bibs/39n2p89.html PMID:10398833

  13. Nanoceria particles as catalytic amplifiers for alkaline phosphatase assays.

    PubMed

    Hayat, Akhtar; Andreescu, Silvana

    2013-11-01

    We propose a novel system to enhance detection sensitivity of alkaline phosphatase (ALP) in electrochemical assays by using nanoceria particles as redox active catalytic amplifiers of ALP signals. The catalytic activity of nanoceria particles attributed to their dual oxidation state Ce(4+)/Ce(3+) and high oxygen mobility enabled oxidation of the products of the ALP-catalyzed reaction. A suite of spectroscopic and electrochemical methods, including UV-vis spectroscopy, transmission electron microscopy (TEM), dynamic light scattering (DLS), and cyclic voltammetry (CV) were used to characterize the interaction of nanoceria with the ALP-generated products. Spectrometric experiments demonstrate change in the oxidation state of nanoceria upon exposure to the hydrolytic products of ALP. Three enzymatically generated products of commonly used ALP substrates were detected at a screen printing electrode surface in the presence of nanoceria. Electrochemical experiments demonstrate signal amplification of the ALP activity assay by nanoceria for all three products, demonstrating remarkable sensitivity of this assay. The assay was optimized with respect to pH and buffer composition. Analytical characterization of the nanoceria-based ALP activity assay was established using a 1-naphthyl phosphate substrate. The proposed strategy can find widespread applications in sensing schemes involving ALP. PMID:24053108

  14. Alkaline phosphatase activity in Zostera noltii hornem. and its contribution to the release of phosphate in the palmones river estuary

    NASA Astrophysics Data System (ADS)

    Hernández, I.; Pérez-Llorens, J. L.; Fernández, J. A.; Niell, F. X.

    Alkaline phosphatase activity (APA) was studied in Zostera noltii Hornem., a sea-grass collected in the Palmones river estuary (southern Spain). The higher activity was found in the leaves, with minor contributions in the stem and the underground parts of the plant. The enzymatic activity showed a two-phase kinetic versus substrate concentration between 5 μM and 25 mM. The influence of some environmental factors important in nature (temperature, pH, salinity, photon irradiance and external phosphate) on the enzymatic activity is discussed. Over an ecophysiological range of these factors, maximum APA was found at 30 0C (22·6 μmol pNP released g dry wt-1 h-1), pH 8·8 (35·6 μmol pNP g dry wt-1 h-1) and salinity 43·8 (27·8 pmol pNP g dry wt-1 h-1). With regard to light, APA and phosphate uptake in shoots were light-saturated and showed similar values for maximum velocity and half-saturation constant. In the range of phosphate concentration tested (0 20 μM), APA was independent of the external phosphate concentration. Finally, as Z. noltii incorporated only 16% of the phosphate hydrolysed from the model phosphomonoester used in the assay, the significance of Z. noltii population in the enzymatic release of phosphate to the estuary was estimated. A minimum of 8·4 nM Pi liberated per day and a maximum of 99·8 nM Pi day-1 was found.

  15. Alkaline Phosphatase Activity in Zostera noltii Hornem. and its Contribution to the Release of Phosphate in the Palmones River Estuary

    NASA Astrophysics Data System (ADS)

    Hernández, I.; Pérez-Llorens, J. L.; Fernández, J. A.; Niell, F. X.

    Alkaline phosphatase activity (APA) was studied in Zostera noltii Hornem., a sea-grass collected in the Palmones river estuary (southern Spain). The higher activity was found in the leaves, with minor contributions in the stem and the underground parts of the plant. The enzymatic activity showed a two-phase kinetic versus substrate concentration between 5 μM and 25 mM. The influence of some environmental factors important in nature (temperature, pH, salinity, photon irradiance and external phosphate) on the enzymatic activity is discussed. Over an ecophysiological range of these factors, maximum APA was found at 30 °C (22·6 μmol pNP released g dry wt -1 h -1), pH 8·8 (35·6 μmol pNP g dry wt -1 h -1) and salinity 43·8 (27·8 μmol pNP g dry wt -1 h -1). With regard to light, APA and phosphate uptake in shoots were light-saturated and showed similar values for maximum velocity and half-saturation constant. In the range of phosphate concentration tested (0-20 μM), APA was independent of the external phosphate concentration. Finally, as Z. noltii incorporated only 16% of the phosphate hydrolysed from the model phosphomonoester used in the assay, the significance of Z. noltii population in the enzymatic release of phosphate to the estuary was estimated. A minimum of 8·4 nM Pi liberated per day and a maximum of 99·8 nM Pi day -1 was found.

  16. Alkaline phosphatase activity in Zostera noltii hornem. and its contribution to the release of phosphate in the palmones river estuary

    NASA Astrophysics Data System (ADS)

    Hernández, I.; Pérez-Llorens, J. L.; Fernández, J. A.; Niell, F. X.

    Alkaline phosphatase activity (APA) was studied in Zostera noltii Hornem., a sea-grass collected in the Palmones river estuary (southern Spain). The higher activity was found in the leaves, with minor contributions in the stem and the underground parts of the plant. The enzymatic activity showed a two-phase kinetic versus substrate concentration between 5 μM and 25 mM. The influence of some environmental factors important in nature (temperature, pH, salinity, photon irradiance and external phosphate) on the enzymatic activity is discussed. Over an ecophysiological range of these factors, maximum APA was found at 30 0C (22·6 μmol pNP released g dry wt -1 h -1), pH 8·8 (35·6 μmol pNP g dry wt -1 h -1) and salinity 43·8 (27·8 pmol pNP g dry wt -1 h -1). With regard to light, APA and phosphate uptake in shoots were light-saturated and showed similar values for maximum velocity and half-saturation constant. In the range of phosphate concentration tested (0-20 μM), APA was independent of the external phosphate concentration. Finally, as Z. noltii incorporated only 16% of the phosphate hydrolysed from the model phosphomonoester used in the assay, the significance of Z. noltii population in the enzymatic release of phosphate to the estuary was estimated. A minimum of 8·4 nM Pi liberated per day and a maximum of 99·8 nM Pi day -1 was found.

  17. 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.

  18. Rock phosphate solubilization under alkaline conditions by Aspergillus japonicus and A. foetidus.

    PubMed

    Singal, R; Gupta, R; Saxena, R K

    1994-01-01

    Aspergillus japonicus and A. foetidus were found to solubilize five types of Indian rock phosphates at pH 8 and 9. Solubilization was higher in the presence of pyrite than in controls lacking either pyrite or fungal inoculum. Both the aspergilli were found to be good pyrite solubilizers and could grow over a wide pH range. Solubilization of rock phosphates was the result of organic acid release and pyrite oxidation.

  19. Biochemical and proteomic characterisation of haemolymph serum reveals the origin of the alkali-labile phosphate (ALP) in mussel (Mytilus galloprovincialis).

    PubMed

    Oliveri, Caterina; Peric, Lorena; Sforzini, Susanna; Banni, Mohammed; Viarengo, Aldo; Cavaletto, Maria; Marsano, Francesco

    2014-09-01

    Mollusc haemolymph proteins are known to play several important physiological roles in the immune system, heavy metal transport and the tissue distribution of lipophilic compounds. In this study, we analysed acetone-extracted proteins from mussel haemolymph by one- and two-dimensional gel electrophoresis. The proteins were identified by comparing mass spectrometry data with the invertebrate EST database, allowing us to establish the mussel haemolymph serum proteome. Extrapallial protein (EP) precursor represents the most abundant serum protein; astacin and CuZn superoxide dismutase were also detected. Slight contamination from muscle proteins, due to the sampling method, was also found. No differences were observed in the profiles obtained for male and female serum proteins. One aspect of interest was the previously reported finding that alkali-labile phosphate (ALP) from haemolymph serum may be representative of vitellogenin (vtg)-like protein content in the circulatory fluid of molluscs. In our analysis of mussel haemolymph serum, vitellogenin-like proteins were never found. To confirm these data, a typical methyl-tert-butyl-ether (MTBE) extraction, which is specific for vtg-like proteins, was performed, and the results of the electrophoretic analyses were compared with those obtained by acetonic precipitation. The results showed that the electrophoretic profiles are similar and that vtg-like proteins cannot be identified. Moreover, the main phosphoprotein present in female and male extracts is EP protein precursor. In addition, agarose gel electrophoresis demonstrates that high-molecular-weight forms of vtg-like proteins are not detectable.

  20. 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.

  1. 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...

  2. 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.

  3. Enhanced osteogenic activity of a poly(butylene succinate)/calcium phosphate composite by simple alkaline hydrolysis.

    PubMed

    Arphavasin, Suphakit; Singhatanadgit, Weerachai; Ngamviriyavong, Patcharee; Janvikul, Wanida; Meesap, Preeyapan; Patntirapong, Somying

    2013-10-01

    Bone engineering offers the prospect of alternative therapies for clinically relevant skeletal defects. Poly(butylene succinate) (PBSu) is a biodegradable and biocompatible polyester which may possess some limitations in clinical use due to its hydrophobicity. In order to overcome these limitations and increase the bioactivity, a simple and convenient surface hydrolysis of PBSu, PBSu/hydroxyapatite and PBSu/β-tricalcium phosphate (TCP) films was performed. The resulting surfaces (i.e., HPBSu, HPBSu/HA and HPBSu/TCP) were tested for their physicochemical property, biocompatibility and osteogenic potency. The results showed that surface hydrolysis significantly increased surface roughness and hydrophilicity of the composites, with the HPBSu/TCP possessing the most pronounced results. All the materials appeared to be biocompatible and supported in vitro growth and osteoblast differentiation of hMSCs, and the alkaline hydrolysis significantly enhanced the hMSC cell proliferation and the osteogenic potency of PBSu/TCP compared with the non-hydrolyzed sample. In conclusion, the HPBSu/TCP possessed better hydrophilicity, biocompatibility and osteogenic potency in vitro, suggesting that this simple and convenient alkaline hydrolysis could be used to augment the biological property of PBSu-based composites for bone engineering in vivo.

  4. Green/red dual fluorescence detection of total protein and alkaline phosphate-conjugated probes on blotting membranes.

    PubMed

    Top, K P; Hatleberg, G; Berggren, K N; Ryan, D; Kemper, C; Haugland, R P; Patton, W F

    2001-03-01

    A two-color fluorescence detection method is described based upon covalently coupling the succinimidyl ester of BODIPY FL-X to proteins immobilized on poly(vinylidene difluoride) (PVDF) membranes, followed by detection of target proteins using the fluorogenic substrate 9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl(DDAO)-phosphate in combination with alkaline-phosphatase-conjugated reporter molecules. This results in all proteins in the profile being visualized as green signal while those detected specifically with the alkaline-phosphatase conjugate appear as red signal. The dichromatic detection system is broadly compatible with a wide range of analytical imaging devices including UV epi- or transilluminators combined with photographic or charge-coupled device (CCD) cameras, xenon-arc sources equipped with appropriate excitation/emission filters, and dual laser gel scanners outfitted with a 473 nm second-harmonic generation or 488 nm argon-ion laser as well as a 633 nm helium-neon or 635 nm diode laser. The dichromatic detection method permits detection of low nanogram amounts of protein and allows for unambiguous identification of target proteins relative to the entire protein profile on a single electroblot, obviating the need to run replicate gels that would otherwise require visualization of total proteins by silver staining and subsequent alignment with chemiluminescent or colorimetric signals generated on electroblots. PMID:11332758

  5. Surface potential and osteoblast attraction to calcium phosphate compounds is affected by selected alkaline hydrolysis processing.

    PubMed

    Smith, I O; Baumann, M J; Obadia, L; Bouler, J-M

    2004-08-01

    This study examines the link(s) between the suspension behavior of calcium deficient apatites (CDAs) and biphasic calcium phosphate (BCP), as measured by the zeta-potential, with respect to both whole bone and osteoblasts. CDA is fabricated by hydrolyzing an acidic CaP such as dicalcium diphosphate dihydrate (DCPD; CaHPO4.2H2O) and has a structure and composition close to bone apatite. Sintering CDA results in the formation of BCP ceramics consisting of mixtures of hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP), with the HA/beta-TCP weight ratio proportional to the Ca/P ratio of CDA. The choice of the base for the DCPD hydrolysis allows various ionic partial substitution of the formed CDA. Na for Ca partial substitution is of interest because of the resulting improvement in mechanical properties of the resulting BCP ceramics and NH4OH was used as a negative control. The zeta-potential was measured for these materials and the stability of the ceramic to bone interaction calculated. zeta-potential values decrease for CDA(NH4OH) versus CDA(NaOH) and increase for BCP(NH4OH) versus BCP(NaOH). While results of these analyses indicate that NH4OH and NaOH processed CDA and BCP will likely yield osteoblast attachment in vivo, differences in the zeta-potentials may explain varying degrees of cell attachment.

  6. Ratiometric electrochemical detection of alkaline phosphatase.

    PubMed

    Goggins, Sean; Naz, Christophe; Marsh, Barrie J; Frost, Christopher G

    2015-01-11

    A novel ferrocene-derived substrate for the ratiometric electrochemical detection of alkaline phosphatase (ALP) was designed and synthesised. It was demonstrated to be an excellent electrochemical substrate for the ALP-labelled enzyme-linked immunosorbent assay (ELISA).

  7. 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

    2015-06-01

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

  8. 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. PMID:20655385

  9. Structural investigation of Eu{sup 2+} emissions from alkaline earth zirconium phosphate

    SciTech Connect

    Hirayama, Masaaki; Sonoyama, Noriyuki; Yamada, Atsuo; Kanno, Ryoji

    2009-04-15

    Eu{sup 2+} doped A{sub 0.5}Zr{sub 2}(PO{sub 4}){sub 3} (A=Ca, Sr, Ba) phosphors with the NASICON structure were synthesized by a co-precipitation method. Their photoluminescent and structural properties were investigated by photoluminescent spectroscopy and powder X-ray Rietveld analysis, which determined two sites for Eu{sup 2+} ions in the host structure, 3a and 3b. The Eu-O bond lengths were increased by changing alkaline earth ions from Ca to Ba, causing Eu{sup 2+} emission bands to shift from blue-green to blue. A correlation was observed between the peak wavelength positions and the Eu-O bond length. The photoluminescent properties are discussed in terms of crystal field strength and nephelauxetic effect, and a schematic diagram of Eu{sup 2+} emissions is proposed for the Eu{sup 2+} doped NASICON phosphor. - Graphical abstract: Eu{sup 2+} doped NASICON structured A{sub 0.5}Zr{sub 2}(PO{sub 4}){sub 3} (A=Ca, Sr, Ba) showed the blue and blue-green colored emissions attributed to 4f{sup 6}5d{sup 1}-4f{sup 7} transitions. The photoluminescent properties are discussed in terms of crystal field strength and nephelauxetic effect using powder X-ray Rietveld analysis.

  10. Novel cyanobacterial bioreporters of phosphorus bioavailability based on alkaline phosphatase and phosphate transporter genes of Anabaena sp. PCC 7120.

    PubMed

    Muñoz-Martín, M Angeles; Mateo, Pilar; Leganés, Francisco; Fernández-Piñas, Francisca

    2011-07-01

    There is heterogeneity in the way cyanobacteria respond to P starvation and subsequently how they adapt to environments with low or fluctuating P concentrations. In this study, we have fused the promoterless lux operon luxCDABE to the promoter regions of Anabaena sp. PCC 7120 phoA genes putatively encoding alkaline phosphatases, phoA (all2843) and phoA-like (alr5291) and to the promoter region of one operon putatively encoding a high affinity phosphate transporter pst1 (all4575-4572). The self-bioluminescent strains constructed in this way, Anabaena AP (phoA promoter), Anabaena AP-L (phoA-like promoter), and Anabaena PST (pst1 promoter) have been used to study the expression of these genes in response to P starvation and P re-feeding with inorganic and organic phosphate sources. Our data showed that the pst1 promoter was activated at much higher level than the phoA-like promoter following P starvation; however, we did not observe activation of the phoA promoter. The P re-feeding experiments revealed that both strains, Anabaena (A.) PST and A. AP-L could be used as novel bioreporters of P availability in environmental samples. Both strains were used to estimate bioavailable P in environmental samples (fresh- and wastewaters) with a wide range of soluble P concentrations. The results indicated that most of the P in the water samples was in chemical forms available to the cyanobacterium; however there were some differences in the estimates given by both strains as A. PST appeared to be more adequate for the samples with the lowest P load while A. AP-L gave similar or even higher values of P concentrations than those chemically measured in samples with higher P load.

  11. Imaging of alkaline phosphatase activity in bone tissue.

    PubMed

    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 (19)Flourine magnetic resonance spectroscopic imaging ((19)FMRSI). 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 (19)Fluorine magnetic resonance spectroscopy ((19)FMRS) and (19)FMRSI. 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. (19)FMRS and (19)FMRSI were applied for the non-invasive measurement of DiFMUP hydrolysis. (19)FMRS 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 (19)FMRS and enabled the development of a pharmacokinetic model of ALP activity. Application of (19)FMRSI facilitated anatomically accurate, non-invasive imaging of ALP concentration and activity in rat bone. Thus, (19)FMRSI 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. 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

  13. 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. PMID:27405261

  14. Alkaline phosphatase induces the mineralization of sheets of collagen implanted subcutaneously in the rat.

    PubMed Central

    Beertsen, W; van den Bos, T

    1992-01-01

    To determine whether alkaline phosphatase (ALP) can cause the mineralization of collagenous matrices in vivo, bovine intestinal ALP was covalently bound to slices of guanidine-extracted demineralized bovine dentin (DDS). The preparations were implanted subcutaneously over the right half of the rat skull. Control slices not treated with the enzyme were implanted over the left half of the skull of the same animals. Specimens were harvested after periods varying from 1 to 4 wk. It was shown that ALP-coupled DDS rapidly accumulated hydroxyapatite crystals. 4 wk after implantation, the content of calcium and phosphate per microgram of hydroxyproline amounted up to 80 and 60%, respectively, of that found in normal bovine dentin. Our observations present direct evidence that ALP may play a crucial role in the induction of hydroxyapatite deposition in collagenous matrices in vivo. Images PMID:1602003

  15. 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. PMID:25476378

  16. 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.

  17. 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. PMID:26666254

  18. 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.

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

    PubMed

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

    2016-10-01

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

  20. 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. PMID:26645431

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

    PubMed Central

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

    2015-01-01

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

  2. AMP/GMP Analogs as Affinity ESIPT Probes for Highly Selective Sensing of Alkaline Phosphatase Activity in Living Systems.

    PubMed

    Jia, Yan; Li, Peng; Han, Keli

    2015-11-01

    Current probes for alkaline phosphatase (ALP) detection had been developed mainly by adding a phosphate group to a dye, which would lead to indistinct performance when implemented in a living system as several phosphatases exist together. In this study, the nucleotides adenosine monophosphate (AMP) and guanosine monophosphate (GMP) were introduced into 2'-(2'-hydroxyphenyl)-benzothiazole-based probes, and highly fluorescent turn-on probes with good selectivity towards ALP over several phosphatases, as well as high affinity and low toxicity were obtained. In the presence of L-phenylalanine, an ALP inhibitor, a strong decrease in fluorescence recovery was observed. These probes allowed for real-time imaging of endogenous ALP activity in living cells as well as in a zebrafish model.

  3. 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

    2014-11-01

    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

  4. 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.

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

    NASA Astrophysics Data System (ADS)

    Simescu, Florica; Idrissi, Hassane

    2008-12-01

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

  6. Oxygen-18 Labeling evidence against a hexacoordinate phosphorus intermediate in the alkaline hydrolysis of ethyl ethylene phosphate

    SciTech Connect

    Gorenstein, D.G.; Taira, K.

    1982-11-03

    The role of the hexacoordinate phosphorus intermediate in the reactions of phosphate esters is elucidated in this report. This report presents /sup 18/O labeling results that argue against the formation of such a species in the hydrolysis of a related five-membered ring phosphate ester, ethyl ethylene phosphate. Results confirm that no oxygen exchange from solvent occurs during the course of the reaction or with starting material or products and that there is 100% P-O cleavage for all products at pH2-15 (other results not reported). In addition, most significantly we have found no evidence under any conditions for formation of a hexacoordinate intermediate.

  7. 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.

  8. Pyridoxamine-5-phosphate enzyme-linked immune mass spectrometric assay substrate for linear absolute quantification of alkaline phosphatase to the yoctomole range applied to prostate specific antigen.

    PubMed

    Florentinus-Mefailoski, Angelique; Marshall, John G

    2014-11-01

    There is a need to measure proteins that are present in concentrations below the detection limits of existing colorimetric approaches with enzyme-linked immunoabsorbent assays (ELISA). The powerful enzyme alkaline phosphatase conjugated to the highly specific bacterial protein streptavidin binds to biotinylated macromolecules like proteins, antibodies, or other ligands and receptors with a high affinity. The binding of the biotinylated detection antibody, with resulting amplification of the signal by the catalytic production of reporter molecules, is key to the sensitivity of ELISA. The specificity and amplification of the signal by the enzyme alkaline phosphatase in ELISA together with the sensitivity of liquid chromatography electrospray ionization and mass spectrometry (LC-ESI-MS) to detect femtomole to picomole amounts of reporter molecules results in an ultrasensitive enzyme-linked immune mass spectrometric assay (ELIMSA). The novel ELIMSA substrate pyridoxamine-5-phosphate (PA5P) is cleaved by the enzyme alkaline phosphatase to yield the basic and hydrophilic product pyridoxamine (PA) that elutes rapidly with symmetrical peaks and a flat baseline. Pyridoxamine (PA) and (13)C PA were both observed to show a linear relationship between log ion intensity and quantity from picomole to femtomole amounts by liquid chromatography-electrospray ionization and mass spectrometry. Four independent methods, (i) internal (13)C isotope PA dilution curves, (ii) internal (13)C isotope one-point calibration, (iii) external PA standard curve, and (iv) external (13)C PA standard curve, all agreed within 1 digit in the same order of magnitude on the linear quantification of PA. Hence, a mass spectrometer can be used to robustly detect 526 ymol of the alkaline phosphatase streptavidin probe and accurately quantify zeptomole amounts of PSA against log linear absolute standard by micro electrospray on a simple ion trap.

  9. 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.

  10. Dynamic Evolution of the LPS-Detoxifying Enzyme Intestinal Alkaline Phosphatase in Zebrafish and Other Vertebrates

    PubMed Central

    Yang, Ye; Wandler, Anica M.; Postlethwait, John H.; Guillemin, Karen

    2012-01-01

    Alkaline phosphatases (Alps) are well-studied enzymes that remove phosphates from a variety of substrates. Alps function in diverse biological processes, including modulating host-bacterial interactions by dephosphorylating the Gram-negative bacterial cell wall component lipopolysaccharide (LPS). In animals, Alps are encoded by multiple genes characterized by either ubiquitous expression (named Alpls for their liver expression, but a key to proper bone mineralization), or their tissue-specific expression, for example in the intestine (Alpi). We previously characterized a zebrafish alpi gene (renamed here alpi.1) that is regulated by Myd88-dependent innate immune signaling and that is required to prevent a host’s excessive inflammatory reactions to its resident microbiota. Here we report the characterization of two new alp genes in zebrafish, alpi.2 and alp3. To understand their origins, we investigated the phylogenetic history of Alp genes in animals. We find that vertebrate Alp genes are organized in three clades with one of these clades missing from the mammals. We present evidence that these three clades originated during the two vertebrate genome duplications. We show that alpl is ubiquitously expressed in zebrafish, as it is in mammals, whereas the other three alps are specific to the intestine. Our phylogenetic analysis reveals that in contrast to Alpl, which has been stably maintained as a single gene throughout the vertebrates, the Alpis have been lost and duplicated multiple times independently in vertebrate lineages, likely reflecting the rapid and dynamic evolution of vertebrate gut morphologies, driven by changes in bacterial associations and diet. PMID:23091474

  11. 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.

  12. 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.

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

    SciTech Connect

    Henthorn, P.S.; Raducha, M.; Edwards, Y.H.; Weiss, M.J.; Slaughter, C.; Lafferty, M.A.; Harris, H.

    1987-03-01

    A cDNA clone for human adult intestinal alkaline phosphatase (ALP) (orthophosphoric-monoester phosphohydrolase (alkaline optimum); EC 3.1.3.1) was isolated from a lambdagt11 expression library. The cDNA insert of this clone is 2513 base pairs in length and contains an open reading frame that encodes a 528-amino acid polypeptide. This deduced polypeptide contains the first 40 amino acids of human intestinal ALP, as determined by direct protein sequencing. Intestinal ALP shows 86.5% amino acid identity to placental (type 1) ALP and 56.6% amino acid identity to liver/bone/kidney ALP. In the 3'-untranslated regions, intestinal and placental ALP cDNAs are 73.5% identical (excluding gaps). The evolution of this multigene enzyme family is discussed.

  14. A real-time fluorescent assay for the detection of alkaline phosphatase activity based on carbon quantum dots.

    PubMed

    Qian, Zhao Sheng; Chai, Lu Jing; Huang, Yuan Yuan; Tang, Cong; Shen, Jia Jia; Chen, Jian Rong; Feng, Hui

    2015-06-15

    A convenient and real-time fluorometric assay with the assistance of copper ions based on aggregation and disaggregation of carbon quantum dots (CQDs) was developed to achieve highly sensitive detection of alkaline phosphatase activity. CQDs and pyrophosphate anions (PPi) 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 copper ions, which results in effective fluorescence quenching. Under the catalytic hydrolysis of ALP, PPi can be rapidly transformed to phosphate ions. Stronger affinity of phosphate ions to copper ions than carboxyl groups is taken advantage of to achieve fluorescence recovery induced by re-dispersion of CQDs in the presence of ALP and PPi. Quantitative evaluation of ALP activity in a broad range from 16.7 to 782.6 U/L with the detection limit of 1.1 U/L can be realized in this way, which endows the assay with high enough sensitivity for practical detection in human serum. This strategy broadens the sensing application of fluorescent CQDs with excellent biocompatibility, and provides an example based on disaggregation in optical probe development.

  15. Alkalosis and Dialytic Clearance of Phosphate Increases Phosphatase Activity: A Hidden Consequence of Hemodialysis

    PubMed Central

    Villa-Bellosta, Ricardo; González-Parra, Emilio; Egido, Jesús

    2016-01-01

    Background Extracellular pyrophosphate is a potent endogenous inhibitor of vascular calcification, which is degraded by alkaline phosphatase (ALP) and generated by hydrolysis of ATP via ectonucleotide pyrophosphatase/phosphodiesterase 1 (eNPP1). ALP activity (as routinely measured in clinical practice) represents the maximal activity (in ideal conditions), but not the real activity (in normal or physiological conditions). For the first time, the present study investigated extracellular pyrophosphate metabolism during hemodialysis sessions (including its synthesis via eNPP1 and its degradation via ALP) in physiological conditions. Methods and Findings 45 patients in hemodialysis were studied. Physiological ALP activity represents only 4–6% of clinical activity. ALP activity increased post-hemodialysis by 2% under ideal conditions (87.4 ± 3.3 IU/L vs. 89.3 ± 3.6 IU/L) and 48% under physiological conditions (3.5 ± 0.2 IU/L vs. 5.2 ± 0.2 IU/L). Pyrophosphate synthesis by ATP hydrolysis remained unaltered post-hemodialysis. Post-hemodialysis plasma pH (7.45 ± 0.02) significantly increased compared with the pre-dialysis pH (7.26 ± 0.02). The slight variation in pH (~0.2 units) induced a significant increase in ALP activity (9%). Addition of phosphate in post-hemodialysis plasma significantly decreased ALP activity, although this effect was not observed with the addition of urea. Reduction in phosphate levels and increment in pH were significantly associated with an increase in physiological ALP activity post-hemodialysis. A decrease in plasma pyrophosphate levels (3.3 ± 0.3 μmol/L vs. 1.9 ± 0.1 μmol/L) and pyrophosphate/ATP ratio (1.9 ± 0.2 vs. 1.4 ± 0.1) post-hemodialysis was also observed. Conclusion Extraction of uremic toxins, primarily phosphate and hydrogen ions, dramatically increases the ALP activity under physiological conditions. This hitherto unknown consequence of hemodialysis suggests a reinterpretation of the clinical value of this parameter

  16. Calcium phosphate nanoparticles are associated with inorganic phosphate-induced osteogenic differentiation of rat bone marrow stromal cells.

    PubMed

    Chen, Xiao-rong; Bai, Jing; Yuan, Shuai-jun; Yu, Cai-xia; Huang, Jian; Zhang, Tian-lan; Wang, Kui

    2015-08-01

    In the present study, we demonstrated that calcium phosphate (CaP) nanoparticles formed in cell culture media were implicated in the process of high inorganic phosphate (Pi) mediated osteogenic differentiation of rat bone marrow stromal cells (BMSCs). Exposure of BMSCs in vitro to high Pi-containing media reduced alkaline phosphatase (ALP) activity and the expressions of osteoblast-specific genes. The sediments of CaP nanoparticles were observed at the cell surface and some of them were concomitantly found inside cells at high Pi concentration. In addition, treatment the cells with pyrophosphate (PPi), an inhibitor of calcium crystal formation, abrogated the ALP activity induced by high Pi, suggesting the contribution of CaP nanoparticles. Moreover, for isolated CaP nanoparticles, there was a trend of conversion from amorphous calcium phosphate to hydroxyapatite with elevated Pi. The particle size of CaP increased and the surface morphology changed from spherical to irregular due to increased concentrations of serum proteins incorporated into CaP nanoparticles. The study demonstrated that those physicochemical properties of CaP nanoparticles played an important role in modulating BMSCs differentiation. Furthermore, the addition of Pi in the osteogenic media resulted in a dose-dependent increase in matrix mineralization, while treatment of the cells with PPi suppressed Pi-induced calcium deposition. The findings indicated that calcium deposition in the matrix partly came from the spontaneous precipitation of CaP nanoparticles. PMID:26111760

  17. Calcium phosphate nanoparticles are associated with inorganic phosphate-induced osteogenic differentiation of rat bone marrow stromal cells.

    PubMed

    Chen, Xiao-rong; Bai, Jing; Yuan, Shuai-jun; Yu, Cai-xia; Huang, Jian; Zhang, Tian-lan; Wang, Kui

    2015-08-01

    In the present study, we demonstrated that calcium phosphate (CaP) nanoparticles formed in cell culture media were implicated in the process of high inorganic phosphate (Pi) mediated osteogenic differentiation of rat bone marrow stromal cells (BMSCs). Exposure of BMSCs in vitro to high Pi-containing media reduced alkaline phosphatase (ALP) activity and the expressions of osteoblast-specific genes. The sediments of CaP nanoparticles were observed at the cell surface and some of them were concomitantly found inside cells at high Pi concentration. In addition, treatment the cells with pyrophosphate (PPi), an inhibitor of calcium crystal formation, abrogated the ALP activity induced by high Pi, suggesting the contribution of CaP nanoparticles. Moreover, for isolated CaP nanoparticles, there was a trend of conversion from amorphous calcium phosphate to hydroxyapatite with elevated Pi. The particle size of CaP increased and the surface morphology changed from spherical to irregular due to increased concentrations of serum proteins incorporated into CaP nanoparticles. The study demonstrated that those physicochemical properties of CaP nanoparticles played an important role in modulating BMSCs differentiation. Furthermore, the addition of Pi in the osteogenic media resulted in a dose-dependent increase in matrix mineralization, while treatment of the cells with PPi suppressed Pi-induced calcium deposition. The findings indicated that calcium deposition in the matrix partly came from the spontaneous precipitation of CaP nanoparticles.

  18. Synthesis of a novel beta-tricalcium phosphate/hydroxyapatite biphasic calcium phosphate containing niobium ions and evaluation of its osteogenic properties.

    PubMed

    Tamai, Masato; Isama, Kazuo; Nakaoka, Ryusuke; Tsuchiya, Toshie

    2007-01-01

    To promote the osteogenic properties of osteoblasts, we synthesized a hydroxyapatite (HAp) with beta-tricalcium phosphate (beta-TCP) biphasic calcium phosphate containing Nb ions (NbTCP/HAp). NbTCP/HAp was prepared by annealing precipitates obtained by coprecipitation of an aqueous solution of Ca(NO(3))(2) and a mixture of (NH(4))(2)HPO(4) and aqueous Nb solution. The precipitates can be regarded as a calcium-deficient HAp, the PO(4) sites of which are partly occupied by Nb ions. NbTCP/HAp was successfully synthesized by thermal decomposition of the precipitates. NbTCP/HAp enhanced the calcification of normal human osteoblasts (NHOst), and the amount of calcified tissue increased in proportion to the Nb ion concentration in the NbTCP/HAp. The alkaline phosphatase (ALP) activity of NHOst was also enhanced by NbTCP/HAp. Because Nb ions significantly enhance the ALP activity of NHOst, calcification by NbTCP/HAp is considered to be due to enhancement of ALP activity induced by Nb ions dissolved from NbTCP/HAp. These results indicate that NbTCP/HAp can be an effective bone repair material. PMID:17380293

  19. Utilizing ultrafiltration to remove alkaline phosphatase from clinical analyzer water.

    PubMed

    Bôle, Julien; Mabic, Stéphane

    2006-01-01

    Alkaline phosphatase (ALP) conjugated to antibodies is often used in enzyme immunoassays (EIAs). These assays are notably sensitive to experimental conditions. A possible source of interference is bacterial ALP, which is released when bacterial contamination occurs in clinical analyzers. Preliminary experiments led to the selection of a detection kit, ALP source, and specific types of tubes for collecting water samples and performing assays. The release of ALP from various strains of bacteria identified in pure water was demonstrated (10-30 x 10(6) cfu/mL released 6-10 microU/microL). It was shown that ultrafiltration is totally efficient in removing ALP from water, while residual ALP activity (2.21 microU/microL after filtration of an ALP solution of 6.22 microU/microL) was observed after filtration using a 0.22-mum filter.

  20. Calcium phosphate nanoparticles carrying BMP-7 plasmid DNA induce an osteogenic response in MC3T3-E1 pre-osteoblasts.

    PubMed

    Hadjicharalambous, Chrystalleni; Kozlova, Diana; Sokolova, Viktoriya; Epple, Matthias; Chatzinikolaidou, Maria

    2015-12-01

    Functionalized calcium phosphate nanoparticles with osteogenic activity were prepared. Polyethyleneimine-stabilized calcium phosphate nanoparticles were coated with a shell of silica and covalently functionalized by silanization with thiol groups. Between the calcium phosphate surface and the outer silica shell, plasmid DNA which encoded either for bone morphogenetic protein 7 (BMP-7) or for enhanced green fluorescent protein was incorporated as cargo. The plasmid DNA-loaded calcium phosphate nanoparticles were used for the transfection of the pre-osteoblastic MC3T3-E1 cells. The cationic nanoparticles showed high transfection efficiency together with a low cytotoxicity. Their potential to induce an osteogenic response by transfection was demonstrated by measuring the alkaline phosphatase (ALP) activity and calcium deposition with alizarin red staining. The expression of the osteogenic markers Alp, Runx2, ColIa1 and Bsp was investigated by means of real-time quantitative polymerase chain reaction. It was shown that phBMP-7-loaded nanoparticles can provide a means of transient transfection and localized production of BMP-7 in MC3T3-E1 cells, with a subsequent increase of two osteogenic markers, specifically ALP activity and calcium accumulation in the extracellular matrix. Future strategies to stimulate bone regeneration focus into enhancing transfection efficiency and achieving higher levels of BMP-7 produced by the transfected cells.

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

    PubMed

    Obayashi, Yusuke; Iino, Ryota; Noji, Hiroyuki

    2015-08-01

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

  2. 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

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

    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

  4. 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 ...

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

    PubMed Central

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

    2016-01-01

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

  6. 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

  7. 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.

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

    PubMed

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

    2015-12-23

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

  9. 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.

  10. 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.

  11. Highly sensitive detection of alkaline phosphatase using molecular beacon probes based on enzymatic polymerization.

    PubMed

    Ma, Changbei

    2012-06-01

    We have developed a new methodology for highly sensitive alkaline phosphatase assay using molecular beacon probes. No incubation step is needed to obtain a limit of detection for ALP of 2×10(-16) M. Furthermore, ALP inhibition by the inhibitor okadaic acid is shown, demonstrating the potential for high-throughput screening for inhibitors.

  12. 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.

  13. 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. PMID:25968262

  14. 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.

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

    PubMed

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

    2011-06-01

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

  16. Multicolor ELISA based on alkaline phosphatase-triggered growth of Au nanorods.

    PubMed

    Li, Yanyan; Ma, Xiaoming; Xu, Zhengming; Liu, Meihua; Lin, Zhenyu; Qiu, Bin; Guo, Longhua; Chen, Guonan

    2016-05-10

    Seed-mediated synthesis of gold nanorods (AuNRs) has been widely used for diverse applications in the past decade. In this work, this synthetic process is demonstrated for multicolor biosensing for the first time. Our investigation reveals that ascorbic acid acts as a key factor to mediate the growth of AuNRs. This phenomenon is incorporated into the alkaline phosphatase (ALP)-enzyme-linked immunosorbent assay (ELISA) system based on the fact that ALP can catalyze the conversion of ascorbic acid-phosphate into ascorbic acid with high efficiency. This allows us to develop a multicolor ELISA approach for sensitive detection of disease biomarkers with the naked eye. We show the proof-of-concept multicolor ELISA for the detection of prostate-specific antigen (PSA) in human serum. The results show that different colors are presented in response to different concentrations of PSA, and a detection limit of 3 × 10(-15) g mL(-1) in human serum was achieved. The proposed multicolor ELISA could be a good supplement to conventional ELISA for POC diagnostics. PMID:27050384

  17. The effects of calcium phosphate particles on the growth of osteoblasts.

    PubMed

    Sun, J S; Tsuang, Y H; Liao, C J; Liu, H C; Hang, Y S; Lin, F H

    1997-12-01

    With advances in ceramics technology, calcium phosphate bioceramics have been applied as bone substitutes for several decades. The focus of this work is to elucidate the biocompatibility of the particulates of various calcium phosphate cytotoxicities. Four different kinds of calcium phosphate powders, including beta-tricalcium phosphate (beta-TCP), hydroxyapatite (HA), beta-dicalcium pyrophosphate (beta-DCP), and sintered beta-dicalcium pyrophosphate (SDCP), were tested by osteoblast cell culture. The results were analyzed by cell count, concentration of transforming growth factor-beta 1 (TGF-beta 1), alkaline phosphatase (ALP), and prostaglandin E2 (PGE2) in culture media. The changes were most significant when osteoblasts were cultured with beta-TCP and HA bioceramics. The changes in cell population of the beta-TCP and HA were quite low in the first 3 days, then increased gradually toward the seventh day. The changes in TGF-beta 1 concentration in culture medium inversely related to the changes in cell population. The ALP titer in the culture media of the beta-TCP and HA were quite high in the first 3 days, then decreased rapidly between the third and seventh days. The concentrations of PGE2 in the culture media tested were quite high on the first day, decreased rapidly to the third day, and then gradually until the seventh day. The changes in the beta-DCP and SDCP were quite similar to those of HA and beta-TCP but much less significant. We conclude that HA and beta-TCP have an inhibitory effect on the growth of osteoblasts. The inhibitins effects of the HA and beta-TCP powders on the osteoblast cell cultures possibly are mediated by the increased synthesis of PGE2.

  18. Silicate-substituted calcium phosphate with enhanced strut porosity stimulates osteogenic differentiation of human mesenchymal stem cells.

    PubMed

    De Godoy, Roberta Ferro; Hutchens, Stacy; Campion, Charlie; Blunn, Gordon

    2015-01-01

    While many synthetic ceramic bone graft substitutes (BGSs) have osteoconductive properties (e.g. provide a physical scaffold for osteointegration of surrounding bone tissue), certain BGSs are osteostimulative in that they actively upregulate mesenchymal stem cell proliferation and stimulate differentiation into osteoblast-like cells. The osteostimulative properties of silicate-substituted calcium phosphate with enhanced porosity (SiCaP EP) were evaluated in vitro with STRO-1+ immunoselected human bone marrow derived mesenchymal stem cells (HBMSCs). Osteostimulative materials (SiCaP) and Bioglass 45S5 (Bioglass) were also assessed as positive controls along with non-silicate substituted hydroxyapatite as a negative control. HBMSCs were also assessed on Thermanox discs cultured in basal and osteogenic media to determine when osteogenic differentiation could be significantly detected with this in vitro cell system. HBMSC viability and necrosis, total DNA content, alkaline phosphatase (ALP) expression, and osteocalcin expression were evaluated after 7, 14, 21, and 28 days. It was demonstrated that SiCaP EP is osteostimulative based on its propensity to support STRO-1+ HBMSC proliferation and ability to promote the differentiation of HBMSCs down the osteoblastic lineage from ALP-expressing, matrix-producing osteoblasts to Osteocalcin-producing pre-osteocytes without the presence of external osteogenic factors. SiCaP EP permitted greater HBMSC attachment as well as ALP and Osteocalcin expression than Bioglass which may be attributed to its microstructure and chemistry.

  19. Structure and expression of rat osteosarcoma (ROS 17/2.8) alkaline phosphatase: product of a single copy gene.

    PubMed Central

    Thiede, M A; Yoon, K; Golub, E E; Noda, M; Rodan, G A

    1988-01-01

    Alkaline phosphatase [ALP; orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] is a ubiquitous enzyme of unknown function expressed at high levels in cells of mineralizing tissues. To study the structure, function, and expression of ALP, a full-length cDNA of rat ALP (2415 bases) was isolated from a ROS 17/2.8 osteosarcoma cell lambda gt10 cDNA library. The predicted amino acid sequence spans 524 residues and includes an N-terminal signal peptide of 17 amino acids, the phosphohydrolase active site, a rather hydrophilic backbone with five potential N-glycosylation sites, and a short hydrophobic C-terminal sequence. ALP negative CHO cells transfected with an expression vector containing the ALP coding sequences express ALP. The rat bone, liver, and kidney ALP shows remarkable 90% homology with the corresponding human enzyme, the most divergent region being the C-terminal hydrophobic domain through which the enzyme may be anchored to the plasma membrane. The rat ALP also shows 50% homology with the human placental and intestinal ALP and 25% homology with the Escherichia coli ALP. The amino acids involved in catalysis show nearly complete homology among all known ALP sequences, suggesting that these enzymes evolved from a common ancestral gene. The rat ALP cDNA pRAP 54, used as a hybridization probe in RNA blot analysis of several tissues that express ALP, revealed the presence of an ALP mRNA of approximately equal to 2500 bases. Furthermore, hybridization patterns derived from Southern blot analysis of rat chromosomal DNA offered molecular evidence that the ALP expressed in ROS 17/2.8 osteosarcoma and various rat tissues, excluding the intestine, is the product of the same single copy gene. Images PMID:3422431

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

    PubMed Central

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

    2014-01-01

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

  1. The Alps and the imagination.

    PubMed

    Fleming, Fergus

    2004-11-01

    In the late 19th century, mountaineer and essayist Sir Leslie Stephen wrote that the Alps were "Europe's Playground." His words were prescient: the Alps are today one of the continent's most valued recreation areas. However, the importance of Stephen's words can only be appreciated when set in historical context. One hundred years earlier, his remark would have been laughed at. Until the late 18th century the Alps were a source of fear to travellers and of mystery to scientists. They were an uncharted wilderness at the heart of the world's most crowded continent. Yet, within a remarkably short space of time, they were mapped, developed and exploited. Their current status as "playground" is bound intrinsically to a shift in imagination, beginning in the 16th century and accelerating dramatically between the years 1800 and 1914, that has transformed them, in popular perception, from a realm of terror and superstition to one of beauty, relaxation and contemplation. Driven by fresh perspectives in the fields of science, music, literature and aesthetics, the metamorphosis has affected our appreciation not only of the Alps but of every mountain range in the world.

  2. Key role of alkaline phosphatase in the development of human-derived nanoparticles in vitro.

    PubMed

    Hunter, Larry W; Shiekh, Farooq A; Pisimisis, George T; Kim, Sung-Hoon; Edeh, Samuel N; Miller, Virginia M; Lieske, John C

    2011-03-01

    Alkaline phosphatase (ALP) is an enzyme critical for physiological and pathological biomineralization. Experiments were designed to determine whether ALP participates in the formation of calcifying nanometer sized particles (NPs) in vitro. Filtered homogenates of human calcified carotid artery, aorta and kidney stones were inoculated into cell culture medium containing 10% fetal bovine serum in the absence or presence of inhibitors of ALP or pyrophosphate. A calcific NP biofilm developed within 1 week after inoculation and their development was reduced by pyrophosphate and inhibitors of ALP. ALP protein and enzymatic activity were detected in washed NPs, whether calcified or decalcified. Therefore, ALP activity is required for the formation of calcifying NPs in vitro, as has previously been implicated during pathological calcification in vivo. PMID:21029794

  3. 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.

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

    PubMed

    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 (PO4 (3-)) and were then exposed to an addition of 4.4 pM free Cd(2+) at mid-log phase and harvested after 24 h. Whereas Zn and PO4 (3-) had little effect on overall growth rates, in the final 24 h of the experiment three growth effects were noticed: (i) low PO4 (3-) treatments showed increased growth rates relative to high PO4 (3-) treatments, (ii) the Zn/high PO4 (3-) treatment appeared to enter stationary phase, and (iii) Cd increased growth rates further in both the low PO4 (3-) and Zn treatments. Global proteomic analysis revealed that: (i) Zn appeared to be critical to the PO4 (3-) response in this organism, (ii) bacterial metallothionein (SmtA) appears correlated with PO4 (3-) stress-associated proteins, (iii) Cd has the greatest influence on the proteome at low PO4 (3-) and Zn, (iv) Zn buffered the effects of Cd, and (v) in the presence of both replete PO4 (3-) 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 PO4 (3-) 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 PO4 (3-) 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 PO4 (3-) response in this cyanobacterium. Alternate ALP PhoX (Ca) was found to be a low abundance protein, suggesting that PhoA (Zn, Mg) may be

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

    PubMed

    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 (PO4 (3-)) and were then exposed to an addition of 4.4 pM free Cd(2+) at mid-log phase and harvested after 24 h. Whereas Zn and PO4 (3-) had little effect on overall growth rates, in the final 24 h of the experiment three growth effects were noticed: (i) low PO4 (3-) treatments showed increased growth rates relative to high PO4 (3-) treatments, (ii) the Zn/high PO4 (3-) treatment appeared to enter stationary phase, and (iii) Cd increased growth rates further in both the low PO4 (3-) and Zn treatments. Global proteomic analysis revealed that: (i) Zn appeared to be critical to the PO4 (3-) response in this organism, (ii) bacterial metallothionein (SmtA) appears correlated with PO4 (3-) stress-associated proteins, (iii) Cd has the greatest influence on the proteome at low PO4 (3-) and Zn, (iv) Zn buffered the effects of Cd, and (v) in the presence of both replete PO4 (3-) 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 PO4 (3-) 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 PO4 (3-) 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 PO4 (3-) response in this cyanobacterium. Alternate ALP PhoX (Ca) was found to be a low abundance protein, suggesting that PhoA (Zn, Mg) may be

  6. Radially and Axially Graded Multizonal Bone Graft Substitutes Targeting Critical-Sized Bone Defects from Polycaprolactone/Hydroxyapatite/Tricalcium Phosphate

    PubMed Central

    Ergun, Asli; Yu, Xiaojun; Valdevit, Antonio; Ritter, Arthur

    2012-01-01

    Repair and regeneration of critical sized defects via the utilization of polymeric bone graft substitutes are challenges. Here, we introduce radially and axially graded multizonal bone graft substitutes fabricated from polycaprolactone (PCL), and PCL biocomposites with osteoconductive particles, that is, hydroxyapatite (HA), and β-tricalcium phosphate (TCP). The novel bone graft substitutes should provide a greater degree of freedom to the orthopedic surgeon especially for repair of critically sized bone defects. The modulus of the graft substitute could be tailored in the axial direction upon the systematic variation of the HA/TCP concentration, while in the radial direction the bone graft substitute consisted of an outer layer with high stiffness, encapsulating a softer core with greater porosity. The biocompatibility of the bone graft substitutes was investigated using in vitro culturing of human bone marrow-derived stromal cells followed by the analysis of cell proliferation and differentiation rates. The characterization of the tissue constructs included the enzymatic alkaline phosphates (ALP) activity, microcomputed tomography imaging, and polymerase chain reaction analysis involving the expressions of bone markers, that is, Runx2, ALP, collagen type I, osteopontin, and osteocalcin, overall demonstrating the differentiation of bone marrow derived stem cells (BMSCs) via osteogenic lineage and formation of mineralized bone tissue. PMID:22764839

  7. 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.

  8. Fabrication of novel poly(lactic acid)/amorphous magnesium phosphate bionanocomposite fibers for tissue engineering applications via electrospinning.

    PubMed

    Zhou, Huan; Nabiyouni, Maryam; Lin, Boren; Bhaduri, Sarit B

    2013-05-01

    Fibrous bionanocomposites consisting of amorphous magnesium phosphate (AMP) nanospheres and polylactic acid (PLA) were fabricated by electrospinning. There are two important signatures of this paper. First, AMP, as an alternative to well-known calcium phosphate (CaP) materials, is added to PLA as the second phase. To the best of our knowledge, it is the first attempt to fabricate magnesium phosphate (MgP)/biopolymer composite. This is made possible by our previously reported research on the successful synthesis of AMP nanospheres via microwave processing. Second, the sustained release of magnesium and phosphate ions from PLA matrix can stimulate a series of cell responses. The structure of the composites and their bone-like apatite-forming abilities in simulated body fluid (SBF) were examined. Additionally, the effects on the proliferation and differentiation of preosteoblast cells were evaluated by performing in vitro cell culture and monitoring markers such as Osteocalcin (OCN), Osteopontin (OPN), Alkaline phosphatase (ALP) and Collagen type-I (Col I) using real-time polymerase chain reaction (PCR). For better dispersion of AMP in the fibers, a surfactant, 12-hydroxysteric acid (HSA), as previously reported in the literature, was used. However, HSA significantly inhibited the proliferation and differentiation of preosteoblast cells, indicating the potential risk in using HSA in the combination of AMP or MgP in tissue engineering applications. PMID:23498262

  9. 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.

  10. Human embryonic stem cell encapsulation in alginate microbeads in macroporous calcium phosphate cement for bone tissue engineering.

    PubMed

    Tang, M; Chen, W; Weir, M D; Thein-Han, W; Xu, H H K

    2012-09-01

    Human embryonic stem cells (hESC) are promising for use in regenerative medicine applications because of their strong proliferative ability and multilineage differentiation capability. To date there have been no reports on hESC seeding with calcium phosphate cement (CPC). The objective of this study was to investigate hESC-derived mesenchymal stem cell (hESCd-MSC) encapsulation in hydrogel microbeads in macroporous CPC for bone tissue engineering. hESC were cultured to form embryoid bodies (EB), and the MSC were then migrated out of the EB. hESCd-MSC had surface markers characteristic of MSC, with positive alkaline phosphatase (ALP) staining when cultured in osteogenic medium. hESCd-MSC were encapsulated in alginate at a density of 1millioncellsml(-1), with an average microbead size of 207μm. CPC contained mannitol porogen to create a porosity of 64% and 218-μm macropores, with 20% absorbable fibers for additional porosity when the fibers degrade. hESCd-MSC encapsulated in microbeads in CPC had good viability from 1 to 21days. ALP gene expression at 21days was 25-fold that at 1day. Osteocalcin (OC) at 21days was two orders of magnitude of that at 1day. ALP activity in colorimetric p-nitrophenyl phosphate assay at 21days was fivefold that at 1day. Mineral synthesis by the encapsulated hESCd-MSC at 21days was sevenfold that at 1day. Potential benefits of the CPC-stem cell paste include injectability, intimate adaptation to complex-shaped bone defects, ease in contouring to achieve esthetics in maxillofacial repairs, and in situ setting ability. In conclusion, hESCd-MSC were encapsulated in alginate microbeads in macroporous CPC, showing good cell viability, osteogenic differentiation and mineral synthesis for the first time. The hESCd-MSC-encapsulating macroporous CPC construct is promising for bone regeneration in a wide range of orthopedic and maxillofacial applications.

  11. Phosphate salts

    MedlinePlus

    ... taken by mouth or used as enemas. Indigestion. Aluminum phosphate and calcium phosphate are FDA-permitted ingredients ... Phosphate salts containing sodium, potassium, aluminum, or calcium are LIKELY SAFE for most people when taken by mouth short-term, when sodium phosphate is inserted into the ...

  12. Human mesenchymal stromal cells response to biomimetic octacalcium phosphate containing strontium.

    PubMed

    Birgani, Zeinab Tahmasebi; Malhotra, Angad; van Blitterswijk, Clemens A; Habibovic, Pamela

    2016-08-01

    The incorporation of bioinorganics into synthetic biomaterials is a promising approach to improve the biological performance of bone graft substitutes, while still retaining their synthetic nature. Among these bioinorganics, strontium ions (Sr(2+) ) have reported enhanced bone formation, and a reduced risk of bone fractures. While previous results have been encouraging, more detailed studies are needed to further develop specific applications. This study demonstrates the effects of Sr(2+) on the osteogenic differentiation of human mesenchymal stromal cells (hMSCs) when introduced as either a dissolved salt, or incorporated into biomimetic calcium phosphate (CaP) coatings. Upon attachment, hMSCs seeded in the presence of higher Sr(2+) concentrations presented with a more elongated shape as compared to the controls without Sr(2+) . Both Sr(2+) as a dissolved salt in the medium, or incorporated into CaP coatings, positively influenced hMSC alkaline phosphatase (ALP) activity in a dose-dependent manner. At the mRNA level, the expression of osteogenic markers ALP, bone sialoprotein, bone morphogenetic protein 2, osteopontin, and osteoclacin were increased in the presence of Sr(2+) , independent of the delivery method. Overall, this study demonstrates the positive effects of strontium on the osteogenic differentiation of human MSCs, and supports the use of strontium-incorporated CaPs for bone regeneration applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1946-1960, 2016. PMID:27012665

  13. Effect of culture conditions and calcium phosphate coating on ectopic bone formation.

    PubMed

    Vaquette, Cédryck; Ivanovski, Saso; Hamlet, Stephen M; Hutmacher, Dietmar W

    2013-07-01

    This study investigated the effect of a calcium phosphate (CaP) coating onto a polycaprolactone melt electrospun scaffold and in vitro culture conditions on ectopic bone formation in a subcutaneous rat model. The CaP coating resulted in an increased alkaline phosphatase activity (ALP) in ovine osteoblasts regardless of the culture conditions and this was also translated into higher levels of mineralisation. A subcutaneous implantation was performed and increasing ectopic bone formation was observed over time for the CaP-coated samples previously cultured in osteogenic media whereas the corresponding non-coated samples displayed a lag phase before bone formation occurred from 4 to 8 weeks post-implantation. Histology and immunohistochemistry revealed bone fill through the scaffolds 8 weeks post-implantation for coated and non-coated specimens and that ALP, osteocalcin and collagen 1 were present at the ossification front and in the bone tissues. Vascularisation in the vicinity of the bone tissues was also observed indicating that the newly formed bone was not deprived of oxygen and nutrients. We found that in vitro osteogenic induction was essential for achieving bone formation and CaP coating accelerated the osteogenic process. We conclude that high cell density and preservation of the collagenous and mineralised extracellular matrix secreted in vitro are factors of importance for ectopic bone formation.

  14. 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).

  15. 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...

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

    Code of Federal Regulations, 2014 CFR

    2014-01-01

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

  17. 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...

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

    Code of Federal Regulations, 2010 CFR

    2010-01-01

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

  19. 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...

  20. 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 ... boot at the northwest end of the Adriatic Sea. The barrier islands across the Gulf of Venice are barely visible. Trieste is on the bay at ...

  1. Effect of strontium ions on calcification of preosteoblasts cultured on porous calcium- and phosphate-containing titanium oxide layers formed by micro-arc oxidation.

    PubMed

    Sato, Mizuki; Chen, Peng; Tsutsumi, Yusuke; Shiota, Makoto; Hanawa, Takao; Kasugai, Shohei

    2016-01-01

    Strontium (Sr) ions were added to calcium- and phosphate-containing porous titanium oxide layers formed by micro-arc oxidation (MAO) of titanium (Ti) substrates to improve their osseointegration. An MC3T3-E1 preosteoblast was used to evaluate the effect of the incorporated Sr species on cell calcification. Similar surface microporous morphologies of the oxide layers were observed for all specimens produced by MAO, while the contents of the incorporated Sr ions increased with increasing Sr concentrations in MAO electrolytes. The calcium- and phosphate-containing porous layers promoted the cell alkaline phosphatase (ALP) activity, while cell calcification was promoted by the Sr addition. In particular, the ALP activity significantly increased after 10 days of culture, and larger areas of calcified deposits were observed for the specimens treated with MAO electrolytes containing 0.15 mol L(-1) of Sr species. The effect of Sr addition on the calcification of the MAO-treated Ti oxide layers was established in this study. PMID:27477229

  2. Osteoblastic Induction on Calcium Phosphate Cement-Chitosan Constructs for Bone Tissue Engineering

    PubMed Central

    Weir, Michael D.; Xu, Hockin H.K.

    2009-01-01

    Calcium phosphate cement (CPC) is osteoconductive and moldable, can conform to complex cavity shapes and set in-situ to form hydroxyapatite. Chitosan could increase the strength and toughness of CPC, but there has been no investigation on recombinant human bone morphogenic protein-2 (rhBMP-2) delivery via CPC-chitosan composite and its effect on osteogenic induction of cells. The objective of this research was to investigate the mechanical properties and osteoblastic induction of MC3T3-E1 cells cultured on CPC containing chitosan and rhBMP-2. Cell viability for CPC with chitosan and BMP was comparable to that of control CPC, while the CPC-chitosan composite was stronger and tougher than CPC control. After 14 d, osteblastic induction was quantified by measuring alkaline phosphatase (ALP) activity. ALP (mean±sd;n=6) of cells seeded on traditional CPC without rhBMP-2 was (143±19) (mM pNpp/min)/(μg DNA). Adding chitosan resulted in an ALP of (161±27). Further addition of rhBMP-2 to the CPC-chitosan composite increased the ALP to (305±111) (p<0.05). All ALP activity on CPC composites was significantly higher when compared to the (10.0±3.3) of tissue culture polystyrene (p<0.05). Flexural strength of CPC containing 15% (mass fraction) chitosan was (19.8±1.4)MPa which more than doubled the (8.0±1.4) MPa of conventional CPC (p < 0.05). The addition of chitosan to CPC increased the fracture toughness from (0.18±0.01)MPa·m1/2 to (0.23±0.02)MPa·m1/2 (p<0.05). The relatively high strength, self-hardening CPC-chitosan composite scaffold is promising as a moderate load-bearing matrix for bone repair, with potential to serve as an injectable delivery vehicle for osteoinductive growth factors to promote osteoblastic induction and bone regeneration. PMID:20166217

  3. Osteoblastic induction on calcium phosphate cement-chitosan constructs for bone tissue engineering.

    PubMed

    Weir, Michael D; Xu, Hockin H K

    2010-07-01

    Calcium phosphate cement (CPC) is osteoconductive and moldable, and it can conform to complex cavity shapes and set in situ to form hydroxyapatite. Chitosan could increase the strength and toughness of CPC, but there has been no investigation on recombinant human bone morphogenic protein-2 (rhBMP-2) delivery via CPC-chitosan composite and its effect on osteogenic induction of cells. The objective of this research was to investigate the mechanical properties and osteoblastic induction of MC3T3-E1 cells cultured on CPC-containing chitosan and rhBMP-2. Cell viability for CPC with chitosan and rhBMP-2 was comparable with that of control CPC, whereas the CPC-chitosan composite was stronger and tougher than CPC control. After 14 days, osteoblastic induction was quantified by measuring alkaline phosphatase (ALP) activity. ALP (mean +/- SD; n = 6) of cells seeded on conventional CPC without rhBMP-2 was (143 +/- 19) (mM pNpp/min)/(mug DNA). The addition of chitosan resulted in an ALP of 161 +/- 27. Further addition of rhBMP-2 to the CPC-chitosan composite increased the ALP to 305 +/- 111 (p < 0.05). All ALP activity on CPC composites was significantly higher when compared with the 10.0 +/- 3.3 of tissue culture polystyrene (p < 0.05). Flexural strength of CPC containing 15% (mass fraction) chitosan was 19.8 +/- 1.4 MPa, which is more than double the 8.0 +/- 1.4 MPa of conventional CPC (p < 0.05). The addition of chitosan to CPC increased the fracture toughness from 0.18 +/- 0.01 MPa.m(1/2) to 0.23 +/- 0.02 MPa.m(1/2) (p < 0.05). The relatively high strength, self-hardening CPC-chitosan composite scaffold is promising as a moderate load-bearing matrix for bone repair, with potential to serve as an injectable delivery vehicle for osteoinductive growth factors to promote osteoblastic induction and bone regeneration. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010. PMID:20166217

  4. 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.

  5. 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

  6. 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.

  7. Human umbilical cord stem cell encapsulation in calcium phosphate scaffolds for bone engineering.

    PubMed

    Zhao, Liang; Weir, Michael D; Xu, Hockin H K

    2010-05-01

    Human bone marrow mesenchymal stem cells (hBMSCs) require an invasive procedure to harvest, and have lower self-renewal potential with aging. Umbilical cord mesenchymal stem cells (hUCMSCs) are a relatively new stem cell source; this study reveals a self-setting and load-bearing calcium phosphate construct that encapsulates these stem cells. The flexural strength (mean+/-sd; n=5) of the hUCMSC-encapsulating calcium phosphate cement (CPC) increased from (3.5+/-1.1) MPa without polyglactin fibers, to (11.7+/-2.1) MPa with 20% of polyglactin fibers (p<0.05). hUCMSCs attached to the bone mineral-mimicking scaffold in the osteogenic media and differentiated down the osteogenic lineage, yielding elevated alkaline phosphatase (ALP) and osteocalcin (OC) gene expressions. ALP and OC on the CPC-fiber scaffold was 2-fold those on CPC control without fibers. hUCMSCs encapsulated inside the scaffolds retained excellent viability and cell density. The encapsulated hUCMSCs inside four different constructs successfully differentiated down the osteogenic lineage and synthesized bone minerals, as confirmed by mineral staining, SEM, and XRD. The percentage of mineral area synthesized by the encapsulated hUCMSCs increased from about 3% at day-7, to 12% at day-21 (p<0.05). In conclusion, this study demonstrated that hUCMSCs encapsulated in the bioengineered scaffolds osteo-differentiated and synthesized bone minerals. The self-setting CPC-chitosan-fiber scaffold supported the viability and osteogenic differentiation of the encapsulated hUCMSCs, and had mechanical strength matching that of cancellous bone.

  8. 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

  9. Influence of platelet-rich plasma on osteogenic differentiation of mesenchymal stem cells and ectopic bone formation in calcium phosphate ceramics.

    PubMed

    Kasten, Philip; Vogel, Julia; Luginbühl, Reto; Niemeyer, Philipp; Weiss, Stefan; Schneider, Sven; Kramer, Martin; Leo, Albrecht; Richter, Wiltrud

    2006-01-01

    Platelet-rich plasma (PRP) contains a mixture of growth factors that play an important role in wound and fracture healing. While PRP enhanced bone formation by autogenous cancellous bone grafts, its influence in combination with different bone substitutes remained unknown. This study evaluated the effect of PRP on osteogenic differentiation and ectopic bone formation of human mesenchymal stem cells (MSC) in distinct resorbable calcium phosphate ceramics. Calcium-deficient hydroxyapatite (CDHA) blocks with a large specific surface area (48 m2/g) and beta-tricalcium phosphate (beta-TCP) with a low specific surface area (<0.5 m2/g) were loaded with 2 x 10(5) bone marrow-derived MSC. Half of the specimens were treated with 5-fold concentrated PRP. Biocomposites were implanted subcutaneously into SCID mice or kept under osteogenic culture conditions for 2 weeks before implantation. The addition of PRP increased the specific alkaline phosphatase (ALP) activity (p = 0.012) in undifferentiated MSC/CDHA composites but not in MSC/beta-TCP composites. Osteogenic preinduction was ineffective for CDHA and reduced ALP activity of beta-TCP composites significantly at explantation. Ectopic bone formation was stronger in MSC/CDHA (7/32) compared to MSC/beta-TCP (2/30) composites, but no influence of PRP was evident. In conclusion, the effect of PRP depended on the type of ceramic and the differentiation status of the MSC, and enhanced ALP activity of MSC on the high surface scaffold CDHA only, but PRP did not improve osteogenesis in our setting.

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

    PubMed

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

    2010-12-01

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

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

    PubMed

    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

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

    PubMed Central

    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. PMID:26436096

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

    PubMed Central

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

    2013-01-01

    Background Chronic kidney disease-mineral bone disorder (CKD-MBD) is a systemic syndrome characterized by imbalances in mineral homeostasis, renal osteodystrophy (ROD) and ectopic calcification. The mechanisms underlying this syndrome in individuals with chronic kidney disease (CKD) are not yet clear. Methods We examined the effect of normal phosphate (NP) or high phosphate (HP) feeding in the setting of CKD on bone pathology, serum biochemistry and vascular calcification in calcification-prone dilute brown non-agouti (DBA/2) mice. Results In both NP and HP-fed CKD mice, elevated serum parathyroid hormone and alkaline phosphatase (ALP) levels were observed, but serum phosphorus levels were equivalent compared with sham controls. CKD mice on NP diet showed trabecular alterations in the long bone consistent with high–turnover ROD, including increased trabecular number with abundant osteoblasts and osteoclasts. Despite trabecular bone and serum biochemical changes, CKD/NP mice did not develop vascular calcification. In contrast, CKD/HP mice developed arterial medial calcification (AMC), more severe trabecular bone alterations and cortical bone abnormalities that included decreased cortical thickness and density, and increased cortical porosity. Cortical bone porosity and trabecular number strongly correlated with the degree of aortic calcification. Conclusions HP feeding was required to induce the full spectrum of CKD-MBD symptoms in CKD mice. PMID:23045434

  14. 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.

  15. 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.

  16. [Phosphate binders].

    PubMed

    Heeb, Rita M

    2016-06-01

    Phosphate binders to treat hyperphosphataemia are part of the medication regime of every dialysis patient. Phosphate binders are taken with every meal (three times a day). Generally, the medication adherence rates of phosphate binders are very low. This is due to inconveniences like their bad taste or their size which makes them hard to swallow. Also nephrologists have differing opinions on phosphate binders as they are aware of the dialysis patients' difficulties to deal with the amount of drugs they are prescribed. Still, phosphate binders are important drugs which have shown potential in reducing mortality by regulating the level of serum phosphate. In order to improve adherence rates, pharmacists have to advise the patients on these drugs' side effects versus the risks associated with omitting their intake. PMID:27439258

  17. Associations between Renal Hyperfiltration and Serum Alkaline Phosphatase

    PubMed Central

    Oh, Se Won; Han, Kum Hyun; Han, Sang Youb

    2015-01-01

    Renal hyperfiltration, which is associated with renal injury, occurs in diabetic or obese individuals. Serum alkaline phosphatase (ALP) level is also elevated in patients with diabetes (DM) or metabolic syndrome (MS), and increased urinary excretion of ALP has been demonstrated in patients who have hyperfiltration and tubular damage. However, little was investigated about the association between hyperfiltration and serum ALP level. A retrospective observational study of the 21,308 adults in the Korea National Health and Nutrition Examination Survey IV-V databases (2008–2011) was performed. Renal hyperfiltration was defined as exceeding the age- and sex-specific 97.5th percentile. We divided participants into 4 groups according to their estimated glomerular filtration rate (eGFR): >120, 90–119, 60–89, and <60 mL/min/1.73 m2. The participants with eGFR >120 mL/min/1.73 m2 showed the highest risk for MS, in the highest ALP quartiles (3.848, 95% CI, 1.876–7.892), compared to the lowest quartile. Similarly, the highest risk for DM, in the highest ALP quartiles, was observed in participants with eGFR >120 ml/min/1.73 m2 (2.166, 95% CI, 1.084–4.329). ALP quartiles were significantly associated with albuminuria in participants with eGFR ≥ 60 ml/min/1.73m2. The highest ALP quartile had a 1.631-fold risk elevation for albuminuria with adjustment of age and sex. (95% CI, 1.158-2.297, P = 0.005). After adjustment, the highest ALP quartile had a 1.624-fold risk elevation, for renal hyperfiltration (95% CI, 1.204–2.192, P = 0.002). In addition, hyperfiltration was significantly associated with hemoglobin, triglyceride, white blood cell count, DM, smoking, and alcohol consumption (P<0.05). The relationship between serum ALP and metabolic disorders is stronger in participants with an upper-normal range of eGFR. Higher ALP levels are significantly associated with renal hyperfiltration in Korean general population. PMID:25853240

  18. Monte Carlo simulations of phosphate polyhedron connectivity in glasses

    SciTech Connect

    ALAM,TODD M.

    2000-01-01

    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.

  19. 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.

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

    PubMed Central

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

    2016-01-01

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

  1. Sub-cellular localisation of alkaline phosphatase activity in the cytoplasm of tammar wallaby (Macropus eugenii) neutrophils and eosinophils.

    PubMed

    Hulme-Moir, K Lisa; Clark, Phillip

    2011-07-15

    Alkaline phosphatase (ALP) has been used in studies of neutrophil morphology and function as a marker for identifying different granule populations. In human neutrophils, ALP is found within secretory vesicles, a rapidly mobilisable vesicle population important for upregulating membrane receptors during early activation. Intra-cellular ALP activity in the heterophils of rabbits and guinea pigs, in contrast, is found only in secondary granules. The neutrophils and eosinophils of tammar wallabies (Macropus eugenii) have previously been reported to contain large amounts of ALP activity when stained using routine cytochemical techniques. To define the subcellular location of ALP in this species, cell suspensions were examined using cerium chloride cytochemistry and transmission electron microscopy (TEM). ALP was found in 2 distinct cytoplasmic compartments. One compartment displayed morphology consistent with a subpopulation of secondary granules while a second tubulo-vesicular population appeared similar to the secretory vesicles of human neutrophils. Thin tubular vesicles containing ALP were also identified within the cytoplasm of tammar wallaby eosinophils. Large numbers of ALP-containing vesicles have not been recognised previously in eosinophils and this may represent a novel cytoplasmic compartment. In both cell types, ALP-containing structures showed alteration in morphology following stimulation with N-formyl-Met-Leu-Phe (fMLP) and PMA. PMID:21596444

  2. A Case of Vitamin D Deficiency without Elevation of Serum Alkaline Phosphatase in a Carrier of Hypophosphatasia.

    PubMed

    Matsuo, Kumihiro; Mukai, Tokuo; Furuya, Akiko; Suzuki, Shigeru; Tanahashi, Yusuke; Azuma, Hiroshi

    2013-10-01

    Elevated serum alkaline phosphatase (ALP) is a screening marker for the diagnosis of vitamin D deficiency, which may fail to be diagnosed if serum ALP is not elevated. Here, we describe a case of vitamin D deficiency without elevation of serum ALP. A 1-year-old Japanese girl was referred to our hospital for the evaluation of genu varum. Her serum intact PTH level was elevated, while her serum ALP level was normal. Furthermore, her serum 25-hydroxyvitamin D level was reduced, and her urine phosphoethanolamine (PEA) level was mildly elevated. ALPL gene analysis revealed she was a heterozygous carrier of hypophosphatasia (c.1559delT). Serum intact PTH and urine PEA evaluations were helpful for diagnosing vitamin D deficiency and hypophosphatasia carrier status, respectively. Therefore, the possibility of vitamin D deficiency without elevation of serum ALP should be considered. PMID:24170964

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

    PubMed Central

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

    2013-01-01

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

  4. Characterization of a Cry1Ac-receptor alkaline phosphatase in susceptible and resistant Heliothis virescens larvae.

    PubMed

    Jurat-Fuentes, Juan L; Adang, Michael J

    2004-08-01

    We reported previously a direct correlation between reduced soybean agglutinin binding to 63- and 68-kDa midgut glycoproteins and resistance to Cry1Ac toxin from Bacillus thuringiensis in the tobacco budworm (Heliothis virescens). In the present work we describe the identification of the 68-kDa glycoprotein as a membrane-bound form of alkaline phosphatase we term HvALP. Lectin blot analysis of HvALP revealed the existence of N-linked oligosaccharides containing terminal N-acetylgalactosamine required for [125I]Cry1Ac binding in ligand blots. Based on immunoblotting and alkaline phosphatase activity detection, reduced soybean agglutinin binding to HvALP from Cry1Ac resistant larvae of the H. virescens YHD2 strain was attributable to reduced amounts of HvALP in resistant larvae. Quantification of specific alkaline phosphatase activity in brush border membrane proteins from susceptible (YDK and F1 generation from backcrosses) and YHD2 H. virescens larvae confirmed the observation of reduced HvALP levels. We propose HvALP as a Cry1Ac binding protein that is present at reduced levels in brush border membrane vesicles from YHD2 larvae. PMID:15265032

  5. 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.

  6. Reduced Levels of Membrane-Bound Alkaline Phosphatase Are Common to Lepidopteran Strains Resistant to Cry Toxins from Bacillus thuringiensis

    PubMed Central

    Jurat-Fuentes, Juan Luis; Karumbaiah, Lohitash; Jakka, Siva Rama Krishna; Ning, Changming; Liu, Chenxi; Wu, Kongming; Jackson, Jerreme; Gould, Fred; Blanco, Carlos; Portilla, Maribel; Perera, Omaththage; Adang, Michael

    2011-01-01

    Development of insect resistance is one of the main concerns with the use of transgenic crops expressing Cry toxins from the bacterium Bacillus thuringiensis. Identification of biomarkers would assist in the development of sensitive DNA-based methods to monitor evolution of resistance to Bt toxins in natural populations. We report on the proteomic and genomic detection of reduced levels of midgut membrane-bound alkaline phosphatase (mALP) as a common feature in strains of Cry-resistant Heliothis virescens, Helicoverpa armigera and Spodoptera frugiperda when compared to susceptible larvae. Reduced levels of H. virescens mALP protein (HvmALP) were detected by two dimensional differential in-gel electrophoresis (2D-DIGE) analysis in Cry-resistant compared to susceptible larvae, further supported by alkaline phosphatase activity assays and Western blotting. Through quantitative real-time polymerase chain reaction (qRT-PCR) we demonstrate that the reduction in HvmALP protein levels in resistant larvae are the result of reduced transcript amounts. Similar reductions in ALP activity and mALP transcript levels were also detected for a Cry1Ac-resistant strain of H. armigera and field-derived strains of S. frugiperda resistant to Cry1Fa. Considering the unique resistance and cross-resistance phenotypes of the insect strains used in this work, our data suggest that reduced mALP expression should be targeted for development of effective biomarkers for resistance to Cry toxins in lepidopteran pests. PMID:21390253

  7. Osteoblastic differentiation under controlled bioactive ion release by silica and titania doped sodium-free calcium phosphate-based glass.

    PubMed

    Shah Mohammadi, Maziar; Chicatun, Florencia; Stähli, Christoph; Muja, Naser; Bureau, Martin N; Nazhat, Showan N

    2014-09-01

    Sodium-free phosphate-based glasses (PGs) doped with both SiO2 and TiO2 (50P2O5-40CaO-xSiO2-(10-x)TiO2, where x=10, 7, 5, 3, and 0mol%) were developed and characterised for controlled ion release applications in bone tissue engineering. Substituting SiO2 with TiO2 directly increased PG density and glass transition temperature, indicating a cross-linking effect of Ti on the glass network which was reflected by significantly reduced degradation rates in an aqueous environment. X-ray diffraction confirmed the presence of Ti(P2O7) in crystallised TiO2-containing PGs, and nuclear magnetic resonance showed an increase in Q(1) phosphate species with increasing TiO2 content. Substitution of SiO2 with TiO2 also reduced hydrophilicity and surface energy. In biological assays, MC3T3-E1 pre-osteoblasts effectively adhered to the surface of PG discs and the incorporation of TiO2, and hence higher stability of the PG network, significantly increased cell viability and metabolic activity indicating the biocompatibility of the PGs. Addition of SiO2 increased ionic release from the PG, which stimulated alkaline phosphatase (ALP) activity in MC3T3-E1 cells upon ion exposure. The incorporation of 3mol% TiO2 was required to stabilise the PG network against unfavourable rapid degradation in aqueous environments. However, ALP activity was greatest in PGs doped with 5-7mol% SiO2 due to up-regulation of ionic concentrations. Thus, the properties of PGs can be readily controlled by modifying the extent of Si and Ti doping in order to optimise ion release and osteoblastic differentiation for bone tissue engineering applications. PMID:24945606

  8. 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.

  9. Western Alps: tectonic model wanted

    NASA Astrophysics Data System (ADS)

    Guillot, F.

    2003-04-01

    Dating the W-Alpine tectono-metamorphic phases was much disputed during the last ten years. After Ar-datings [1] other methods [2, 3] have yielded Eocene instead of Upper Cretaceous ages for HP- and UHP-metamorphism. Rejecting the Eo-Alpine Upper Cretaceous Ar-ages imposes to re-consider the related tectonic models. Paleogeographic differences between Internal and External realms [4] fit with fission-track ages of uplift [5, 6]: the W-Alps would comprise 3 amalgamated orogens, with uplift age <8 Ma (External), 22--30 Ma (Penninic) and ˜33 Ma (Sesia). The 2 latter figures represent lower limits to UHP-age but fixing an upper limit is hampered by the poorly known stratigraphical age of protoliths. From this respect, the presence of pre-metamorphic Eocene sediments in the HP-Middle Penninic realm is no longer a radical difference with UHP-massifs where the stratigraphic record is poor. The Middle Penninic Vanoise shows the best preserved Meso-Cenozoic cover, together with mineral associations of respectably high pressure (up to 6--8 kbars). The Vanoise cover formations have recorded an quiet marine environment, with a growing water depth from Late Jurassic to Eocene times. The cover is locally overturned in the Sauvire massif. There, its top of Eocene age is welded to the Palaeozoic basement. The late- to post-Eocene deposits that seal the contact have also endured Alpine HP-LT events, marked by a prominent W-dipping cleavage. So, the first Alpine event was a turbidite and olistolithe emplacement that occurred not before the Middle Eocene, followed by subduction, itself possibly W-directed. A tentative scenario is (1) Late Eocene emplacement of a turbiditic mélange, topped by the allochthonous Schistes Lustrés over the detached cover, in a deep trough context (2a) short-lived W- to N-dipping subduction down to 18--25 km -- (2b) fast upward exhumation to upper crustal levels, roughly parallel to previous subduction planes -- (3) long-lasting (Late Oligocene

  10. 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.

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

    PubMed

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

    2012-10-01

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

  12. Clinical refinement of the automatic lung parameter estimator (ALPE).

    PubMed

    Thomsen, Lars P; Karbing, Dan S; Smith, Bram W; Murley, David; Weinreich, Ulla M; Kjærgaard, Søren; Toft, Egon; Thorgaard, Per; Andreassen, Steen; Rees, Stephen E

    2013-06-01

    The automatic lung parameter estimator (ALPE) method was developed in 2002 for bedside estimation of pulmonary gas exchange using step changes in inspired oxygen fraction (FIO₂). Since then a number of studies have been conducted indicating the potential for clinical application and necessitating systems evolution to match clinical application. This paper describes and evaluates the evolution of the ALPE method from a research implementation (ALPE1) to two commercial implementations (ALPE2 and ALPE3). A need for dedicated implementations of the ALPE method was identified: one for spontaneously breathing (non-mechanically ventilated) patients (ALPE2) and one for mechanically ventilated patients (ALPE3). For these two implementations, design issues relating to usability and automation are described including the mixing of gasses to achieve FIO₂ levels, and the automatic selection of FIO₂. For ALPE2, these improvements are evaluated against patients studied using the system. The major result is the evolution of the ALPE method into two dedicated implementations, namely ALPE2 and ALPE3. For ALPE2, the usability and automation of FIO₂ selection has been evaluated in spontaneously breathing patients showing that variability of gas delivery is 0.3 % (standard deviation) in 1,332 breaths from 20 patients. Also for ALPE2, the automated FIO2 selection method was successfully applied in 287 patient cases, taking 7.2 ± 2.4 min and was shown to be safe with only one patient having SpO₂ < 86 % when the clinician disabled the alarms. The ALPE method has evolved into two practical, usable systems targeted at clinical application, namely ALPE2 for spontaneously breathing patients and ALPE3 for mechanically ventilated patients. These systems may promote the exploration of the use of more detailed descriptions of pulmonary gas exchange in clinical practice.

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

    PubMed

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

    2016-07-01

    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. 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.

  15. 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.

  16. Alkaline "Permanent" Paper.

    ERIC Educational Resources Information Center

    Pacey, Antony

    1991-01-01

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

  17. Effect of reinforcement particle size on in vitro behavior of beta-tricalcium phosphate-reinforced high-density polyethylene: a novel orthopedic composite.

    PubMed

    Homaeigohar, S S H; Shokrgozar, M A; Javadpour, J; Khavandi, A; Sadi, A Yari

    2006-07-01

    Beta-tricalcium phosphate-reinforced high-density polyethylene (beta-TCP/HDPE) is a new biomaterial, which was made to simulate bone composition and study its capacity to act like bony tissues. This material was produced by replacing mineral component and collagen soft tissue of bone with beta-TCP and HDPE, respectively. The biocompatibility of composite samples with different volume fractions of TCP (20, 30, and 40 vol %) and two different particle sizes (80-100 and 120-140 mesh size) was examined in vitro using the osteoblast cell line G-292 by proliferation, alkaline phosphatase (ALP) production, and cell adhesion assays. Cell-material interaction on the surface of the composites was observed by scanning electron microscopy (SEM). The effect of beta-TCP particle size on behavior of the osteoblast cell line was compared between two groups of the composite samples containing smaller and larger reinforcement particle sizes as well as with those of a negative control. In general, results showed that the composite samples containing larger particles supported a higher rate of proliferation and ALP production by osteoblast cells after 3, 7, and 14 days of incubation compared to the composite samples with smaller particle size and control. Furthermore, more cells were attached to the surface of composite samples containing larger particle size when compared to the smaller particle size composites (p<0.05). This number was nearly equal with numbers adhered on negative control [tissue culture polystyrene (TPS)] and significantly higher in comparison with composite control [polyethylene (PE)] (p<0.05). Adhered cells presented a normal morphology by SEM and many of the cells were seen to be undergoing cell division. These findings indicate that beta-TCP/HDPE composites are biocompatible, nontoxic, and in some cases, act to stimulate proliferation of the cells, ALP production, and cell adhesion when compared to the control counterparts. Furthermore, beta-TCP/HDPE samples

  18. 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. PMID:25996812

  19. A high-resolution, fluorescence-based method for localization of endogenous alkaline phosphatase activity.

    PubMed

    Cox, W G; Singer, V L

    1999-11-01

    We describe a high-resolution, fluorescence-based method for localizing endogenous alkaline phosphatase in tissues and cultured cells. This method utilizes ELF (Enzyme-Labeled Fluorescence)-97 phosphate, which yields an intensely fluorescent yellow-green precipitate at the site of enzymatic activity. We compared zebrafish intestine, ovary, and kidney cryosections stained for endogenous alkaline phosphatase using four histochemical techniques: ELF-97 phosphate, Gomori method, BCIP/NBT, and naphthol AS-MX phosphate coupled with Fast Blue BB (colored) and Fast Red TR (fluorescent) diazonium salts. Each method localized endogenous alkaline phosphatase to the same specific sample regions. However, we found that sections labeled using ELF-97 phosphate exhibited significantly better resolution than the other samples. The enzymatic product remained highly localized to the site of enzymatic activity, whereas signals generated using the other methods diffused. We found that the ELF-97 precipitate was more photostable than the Fast Red TR azo dye adduct. Using ELF-97 phosphate in cultured cells, we detected an intracellular activity that was only weakly labeled with the other methods, but co-localized with an antibody against alkaline phosphatase, suggesting that the ELF-97 phosphate provided greater sensitivity. Finally, we found that detecting endogenous alkaline phosphatase with ELF-97 phosphate was compatible with the use of antibodies and lectins. (J Histochem Cytochem 47:1443-1455, 1999)

  20. 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.

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

    PubMed

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

    2016-07-01

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

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

    PubMed

    Lin, Ya-Ting; Liang, Chenju

    2015-10-01

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

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

    PubMed

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

    2001-01-01

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

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

    PubMed

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

    2014-09-01

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

  5. Ethnic differences in pre-adipocyte intracellular lipid accumulation and alkaline phosphatase activity.

    PubMed

    Ali, Aus T; Chirambo, George; Penny, Clement; Paiker, Janice E; Ikram, Faisel; Psaras, George; Crowther, Nigel J

    2015-01-01

    Alkaline phosphatase (ALP) increases lipid accumulation in human pre-adipocytes. This study was performed to assess whether ethnic differences in the prevalence of obesity in African and European females are related to differences in pre-adipocyte lipid accretion and ALP activity. Pre-adipocytes were isolated from 13 black and 14 white females. Adipogenesis was quantified using the lipid dye, Oil red O, whilst ALP activity was assayed in cell extracts on day zero and 12days after initiating adipogenesis. Lipid levels (OD units/mg protein) were lower in pre-adipocytes from white than black females on day 0 (0.36±0.05 versus 0.44±0.03, respectively; p<0.0005) and day 12 (1.18±0.14 versus 1.80±0.22, respectively; p<0.0005), as was ALP activity (mU/mg protein) on day zero (36.5±5.8 versus 136.4±10.9, respectively; p<0.0005) and day 12 (127±16 versus 278±27, respectively; p<0.0005). Treatment of pre-adipocytes with histidine, an ALP inhibitor, blocked lipid accumulation. Thus, lipid uptake is higher in pre-adipocytes isolated from black compared to white females which parallels the obesity prevalence rates in these population groups. The reason for higher fat accumulation in pre-adipocytes isolated from black females may be related to higher ALP activity.

  6. 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. PMID:26663676

  7. Electrospun gelatin/poly(ε-caprolactone) fibrous scaffold modified with calcium phosphate for bone tissue engineering.

    PubMed

    Rajzer, Izabella; Menaszek, Elżbieta; Kwiatkowski, Ryszard; Planell, Josep A; Castano, Oscar

    2014-11-01

    In this study gelatin (Gel) modified with calcium phosphate nanoparticles (SG5) and polycaprolactone (PCL) were used to prepare a 3D bi-layer scaffold by collecting electrospun PCL and gelatin/SG5 fibers separately in the same collector. The objective of this study was to combine the desired properties of PCL and Gel/SG5 in the same scaffold in order to enhance mineralization, thus improving the ability of the scaffold to bond to the bone tissue. The scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and the wide angle X-ray diffraction (WAXD) measurements confirmed that SG5 nanoparticles were successfully incorporated into the fibrous gelatin matrix. The composite Gel/SG5/PCL scaffold exhibited more enhanced mechanical properties than individual Gel and Gel/SG5 scaffolds. The presence of SG5 nanoparticles accelerated the nucleation and growth of apatite crystals on the surface of the composite Gel/SG5/PCL scaffold in simulated body fluid (SBF). The osteoblast response in vitro to developed electrospun scaffolds (PCL and Gel/SG5/PCL) was investigated by using normal human primary NHOst cell lines. NHOst cell culture studies showed that higher alkaline phosphatase (ALP) activity and better mineralization were obtained in the case of composite materials than in pure PCL scaffolds. The mechanically strong PCL scaffold served as a skeleton, while the Gel/SG5 fibers facilitated cell spreading and mineralization of the scaffold. PMID:25280695

  8. Electrospun gelatin/poly(ε-caprolactone) fibrous scaffold modified with calcium phosphate for bone tissue engineering.

    PubMed

    Rajzer, Izabella; Menaszek, Elżbieta; Kwiatkowski, Ryszard; Planell, Josep A; Castano, Oscar

    2014-11-01

    In this study gelatin (Gel) modified with calcium phosphate nanoparticles (SG5) and polycaprolactone (PCL) were used to prepare a 3D bi-layer scaffold by collecting electrospun PCL and gelatin/SG5 fibers separately in the same collector. The objective of this study was to combine the desired properties of PCL and Gel/SG5 in the same scaffold in order to enhance mineralization, thus improving the ability of the scaffold to bond to the bone tissue. The scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and the wide angle X-ray diffraction (WAXD) measurements confirmed that SG5 nanoparticles were successfully incorporated into the fibrous gelatin matrix. The composite Gel/SG5/PCL scaffold exhibited more enhanced mechanical properties than individual Gel and Gel/SG5 scaffolds. The presence of SG5 nanoparticles accelerated the nucleation and growth of apatite crystals on the surface of the composite Gel/SG5/PCL scaffold in simulated body fluid (SBF). The osteoblast response in vitro to developed electrospun scaffolds (PCL and Gel/SG5/PCL) was investigated by using normal human primary NHOst cell lines. NHOst cell culture studies showed that higher alkaline phosphatase (ALP) activity and better mineralization were obtained in the case of composite materials than in pure PCL scaffolds. The mechanically strong PCL scaffold served as a skeleton, while the Gel/SG5 fibers facilitated cell spreading and mineralization of the scaffold.

  9. 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.

  10. 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.

  11. Mesenchymal stem cell proliferation and differentiation on an injectable calcium phosphate-chitosan composite scaffold.

    PubMed

    Moreau, Jennifer L; Xu, Hockin H K

    2009-05-01

    Calcium phosphate cement (CPC) can be molded or injected to form a scaffold in situ, has excellent osteoconductivity, and can be resorbed and replaced by new bone. However, its low strength limits CPC to non-stress-bearing repairs. Chitosan could be used to reinforce CPC, but mesenchymal stem cell (MSC) interactions with CPC-chitosan scaffold have not been examined. The objective of this study was to investigate MSC proliferation and osteogenic differentiation on high-strength CPC-chitosan scaffold. MSCs were harvested from rat bone marrow. At CPC powder/liquid (P/L) mass ratio of 2, flexural strength (mean+/-sd; n=5) was (10.0+/-1.1) MPa for CPC-chitosan, higher than (3.7+/-0.6) MPa for CPC (p<0.05). At P/L of 3, strength was (15.7+/-1.7)MPa for CPC-chitosan, higher than (10.2+/-1.8)MPa for CPC (p<0.05). Percentage of live MSCs attaching to scaffolds increased from 85% at 1 day to 99% at 14 days. There were (180+/-37) cells/mm(2) on scaffold at 1 day; cells proliferated to (1808+/-317) cells/mm(2) at 14 days. SEM showed MSCs with healthy spreading and anchored on nano-apatite crystals via cytoplasmic processes. Alkaline phosphatase activity (ALP) was (557+/-171) (pNPP mM/min)/(microg DNA) for MSCs on CPC-chitosan, higher than (159+/-47) on CPC (p<0.05). Both were higher than (35+/-32) of baseline ALP for undifferentiated MSCs on tissue-culture plastic (p<0.05). In summary, CPC-chitosan scaffold had higher strength than CPC. MSC proliferation on CPC-chitosan matched that of the FDA-approved CPC control. MSCs on the scaffolds differentiated down the osteogenic lineage and expressed high levels of bone marker ALP. Hence, the stronger CPC-chitosan scaffold may be useful for stem cell-based bone regeneration in moderate load-bearing maxillofacial and orthopedic applications.

  12. Alp7/TACC-Alp14/TOG generates long-lived, fast-growing MTs by an unconventional mechanism

    PubMed Central

    Hussmann, Frauke; Drummond, Douglas R.; Peet, Daniel R.; Martin, Douglas S.; Cross, Robert A.

    2016-01-01

    Alp14 is a TOG-family microtubule polymerase from S. pombe that tracks plus ends and accelerates their growth. To interrogate its mechanism, we reconstituted dynamically unstable single isoform S. pombe microtubules with full length Alp14/TOG and Alp7, the TACC-family binding partner of Alp14. We find that Alp14 can drive microtubule plus end growth at GTP-tubulin concentrations at least 10-fold below the usual critical concentration, at the expense of increased catastrophe. This reveals Alp14 to be a highly unusual enzyme that biases the equilibrium for the reaction that it catalyses. Alp7/TACC enhances the effectiveness of Alp14, by increasing its occupancy. Consistent with this, we show in live cells that Alp7 deletion produces very similar MT dynamics defects to Alp14 deletion. The ability of Alp7/14 to accelerate and bias GTP-tubulin exchange at microtubule plus ends allows it to generate long-lived, fast-growing microtubules at very low cellular free tubulin concentrations. PMID:26864000

  13. 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.

  14. Alkaline phosphatase isoenzyme activities in rheumatoid arthritis: hepatobiliary enzyme dissociation and relation to disease activity.

    PubMed Central

    Aida, S

    1993-01-01

    OBJECTIVES--Hyperphosphatasaemia has been observed occasionally in patients with rheumatoid arthritis (RA), and it has been suggested that the serum alkaline phosphatase (ALP) level is related to the activity of the disease. Therefore, the relationship between serum ALP and RA was studied. METHODS--The serum activities of hepatobiliary enzymes (ALP isoenzymes, gamma-glutamyltranspeptidase (GTP), leucine aminopeptidase (LAP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT)), immunoglobulins, RA haemagglutinin test (RAHA), C reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were observed in 288 patients with rheumatoid arthritis. RESULTS--Serum biliary ALP (ALP1) activity was detected in 31.6% of the patients. In patients positive for ALP1 the respective values of total ALP (ALPt) (p < 0.001), liver ALP (ALP2) (p < 0.001), bone ALP (ALP3) (p < 0.05), gamma-GTP (p < 0.001), LAP (p < 0.001), immunoglobulins IgG (p < 0.01), IgA (p < 0.01), and IgM (p < 0.01), RAHA (p < 0.001), CRP (p < 0.001), ESR (p < 0.001), and articular index (p < 0.001) were significantly higher than in patients who did not have ALP1. Significant Spearman's rank correlations (rs) were demonstrated between serum ALP2 level and the respective values of ALPt (rs = 0.9128, p < 0.001), ALP1 (rs = 0.4443, p < 0.001), ALP3 (rs = 0.5898, p < 0.001), gamma-GTP (rs = 0.2903, p < 0.001), LAP (rs = 0.3093, p < 0.001), IgA (rs = 0.2299, p < 0.01), IgM (rs = 0.1773, p < 0.05), RAHA (rs = 0.2420, p < 0.01), CRP (rs = 0.3532, p < 0.001), ESR (rs = 0.4006, p < 0.001). the articular index (rs = 0.4006, p < 0.001). However, no significant difference or correlation was noted for either AST or ALT. In many patients who showed abnormal hyperphosphatasaemia, hepatobiliary enzyme dissociation was observed: levels of ALPt (in 12.8%), ALP1 (in 31.6%), ALP2 (18.8%), gamma-GTP (in 4.3%), and LAP (in 19.3%) were abnormally high, but both AST and ALT were within normal limits. CONCLUSION

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

    PubMed

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

    2016-09-01

    Metabolic bone disease of prematurity is a condition characterized by reduction in bone mineral content (osteopenia). It is a problem faced by very low birth weight (VLBW) infants because of lack of fetal mineralization during the last trimester. Our aim was to assess serum alkaline phosphatase (ALP) level as an early biomarker for osteopenia in premature infants and to estimate an optimal cutoff value of serum ALP at which osteopenia is detected radiologically in premature newborns.This prospective study was conducted on a cohort of 120 newborn infants of both sex of ≤34 weeks' gestational age and <1500 g birth weight. Two blood samples, from each infant on at least 2 consecutive weeks, were reported for calcium, phosphorus, and ALP. Evidence of osteopenia was evaluated radiologically by performing wrist/knee x-ray.Sixteen infants (13.3%) had evidence of osteopenia in x-ray, whereas 104 infants (86.7%) were nonosteopenic and all the osteopenic infants were <1000-g birth weight. Birth weight and gestational age were significantly inversely related to serum ALP levels. Both samples showed statistically significantly higher mean ALP level in osteopenic than nonosteopenics (P < 0.001, and P < 0.001 respectively). There was no constant value of serum ALP related to radiologic evidence of osteopenia. However, the optimal cutoff value of serum ALP at which osteopenia is detected is 500 IU/L with 100% sensitivity and 80.77% specificity.High levels of ALP can be considered a reliable biomarker to predict the status of bone mineralization and the need for radiological evaluation in premature infants particularly those <1000-g birth weight and <32 weeks' gestation. PMID:27631238

  16. 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.

  17. 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

  18. Plastic responses of larval mass and alkaline phosphatase to cadmium in the gypsy moth larvae.

    PubMed

    Vlahović, Milena; Lazarević, Jelica; Perić-Mataruga, Vesna; Ilijin, Larisa; Mrdaković, Marija

    2009-05-01

    Biochemical analyses can point to toxicant presence before its effects can be detected at higher organizational levels. We investigated responses of larval mass and alkaline phosphatase (ALP) to different cadmium treatments in 4th instar gypsy moth larvae from 20 full-sib families. Changes in trait values and trait plasticities as well as their variation were monitored after acute and chronic exposure or recovery from two cadmium concentrations (Cd(1)=10microg and Cd(2)=30microg Cd/g dry food). Larval mass only decreased, without returning to the control level at recovery stage following chronic cadmium challenge. Acute stress did not change trait value but increased genetic variance of larval mass. Significant ALP activity changes, sensitivity of isozyme patterns (Mr of 60, 64, and 85kDa) and increased variation in ALP plasticity during acute exposure to cadmium point to its possible aplication as an exposure biomarker.

  19. Alp-Water-Scarce - Water Management Strategies Against Water Scarcity in the Alps

    NASA Astrophysics Data System (ADS)

    de Jong, C.

    2009-04-01

    Alp-Water-Scarce is a European project on Water Management Strategies against Water Scarcity in the Alps funded by the Alpine Space programme as part of the "European Territorial Cooperation" scheme. The main challenges of this project are to create local Early Warning Systems against Water Scarcity in the Alps. This system will be based on sound, long-term monitoring and modeling and anchored strongly and actively within a Stakeholder Interaction Forum linked across comparative and contrasting regions across the Alps. The Early Warning System is based on the linkage and improvement of field monitoring and assemblage of qualitative and quantitative data derived both from natural water reservoirs as well as from anthropogenic water use in 28 selected pilot regions selected in France, Italy, Austria, Slovenia and Switzerland. These range across different altitudinal and climatological zones, from humid to semi-arid and include inner alpine dry valleys as well as pre-alpine regions. Both groundwater and surface water systems will be considered in addition to a wide range of regimes from glaciated, snow-fed to karstic. The objectives are to improve water management at the short term (annual scale) and long term (using future scenarios) based on modelling and application of climate change and anthropogenic scenarios. Innovative measures of mitigation and adaptation should predict and prevent future water shortages. Awareness raising and stakeholder interaction will form an important part of problem identification, participation in the project, dissemination of results and implementation of new approaches. Alp-Water-Scarce started in October 2008 for a total period of 3 years and has a global budget of approx. 4 Mill. €. It has 17 project partners including 4 Austrian, 2 Swiss, 3 French, 5 Italian, and 3 Slovenian from local governments, provinces, federal institutes and offices, universities, regional agencies, alpine societies, geological surveys, and chambers of

  20. A signal "on" photoelectrochemical biosensor for assay of protein kinase activity and its inhibitor based on graphite-like carbon nitride, Phos-tag and alkaline phosphatase.

    PubMed

    Yin, Huanshun; Sun, Bing; Dong, Linfeng; Li, Bingchen; Zhou, Yunlei; Ai, Shiyun

    2015-02-15

    A highly sensitive and selective photoelectrochemical (PEC) biosensor is fabricated for the detection of protein kinase activity based on visible-light active graphite-like carbon nitride (g-C3N4) and the specific recognition utility of Phos-tag for protein kinase A (PKA)-induced phosphopeptides. For assembling the substrate peptides, g-C3N4 and gold nanoparticles (g-C3N4-AuNPs) complex is synthesized and characterized. When the immobilized peptides on g-C3N4-AuNPs modified ITO electrode are phosphorylated under PKA catalysis, they can be specifically identified and binded with biotin functionalized Phos-tag (Phos-tag-biotin) in the presence of Zn(2+). Then, through the specific interaction between biotin and avidin, avidin functionalized alkaline phosphatase (avidin-ALP) is further assembled to catalyze its substrate of l-ascorbic acid-2-phosphate trisodium salt (AAP) to produce electron donor of ascorbic acid (AA), resulting an increased photocurrent compared with the absence of phosphorylation event. Based on the specific identification effect of Phos-tag, the fabricated biosensor presents excellent selectivity for capturing the phosphorylated serine residues in the substrate peptides. With the good photoactivity of g-C3N4 and ALP-catalyzed signal amplification, the fabricated biosensor presents high sensitivity and low detection limit (0.015 unit/mL, S/N = 3) for PKA. The applicability of this PEC biosensor is further testified by the evaluation of PKA inhibition by HA-1077 with the IC50 value of 1.18μM. This new strategy is also successfully applied to detect the change of PKA activity in cancer cell lysate with and without drug stimulation. Therefore, the developed PEC method has great potential in screening of kinase inhibitors and highly sensitive detection of kinase activity.

  1. Stimulation of Odontogenesis and Angiogenesis via Bioactive Nanocomposite Calcium Phosphate Cements Through Integrin and VEGF Signaling Pathways.

    PubMed

    Lee, Sang-Im; Lee, Eui-Suk; El-Fiqi, Ahmed; Lee, So-Youn; Eun-Cheol Kim; Kim, Hae-Won

    2016-05-01

    Formulating self-setting calcium phosphate cements (CPCs) with secondary phases particularly in the nanoscale order holds great promise to improve biological properties. Here, we focus on the effect that bioactive glass nanoparticles (BGN) incorporated in CPC compositions can have on the proliferation, odontogenic differentiation, and angiogenic stimulation of stem cells derived from human dental pulp (HDPSCs). These odontogenic and angiogenic events are of special importance in the dentin-pulp regeneration processes. In comparison to pure CPCs, nanocomposite cements exhibit a significantly improved proliferation of HDPSCs, and the improvement is more significant as the BGN content increases. The nanocomposite cements substantially enhance the adhesion of cells, and significantly up-regulate odontogenic differentiation, including alkaline phosphatase (ALP) activity and the expressions of odontogenic genes (sialophosphoprotein, dentin matrix protein I, ALP, osteopontin and osteocalcin). Furthermore, the use of nanocomposite cements result in stimulation of angiogenic gene expression (VEGF, FGF-2, VEGFRs, PECAM-1, and VE-cadherin) and protein production (VEGF, VEGFR-1). The angiogenic stimulation by the HDPSCs significantly affects the endothelial cell behaviors, that is, the endothelial cell migration and the tubular network formation are substantially improved when treated with HDPSC-conditioned medium, particularly with the help of nanocomposite cements. The integrin and VEGF signaling pathways are reasoned for the stimulation of the odontogenesis and angiogenesis of cells, where the nanocomposite cements up-regulate the integrin subsets α1, α2, α3, and β1, and activate the integrin downstream signal pathways, such as p-FAK, p-Akt, p-paxillin, JNK, EK, and NF-κB, as well as other nuclear transcriptional factors, including CREB, STAT-3, and ELK-1. The current results indicate that the new formulation of the nanocomposite self-setting cements might provide some

  2. Stimulation of Odontogenesis and Angiogenesis via Bioactive Nanocomposite Calcium Phosphate Cements Through Integrin and VEGF Signaling Pathways.

    PubMed

    Lee, Sang-Im; Lee, Eui-Suk; El-Fiqi, Ahmed; Lee, So-Youn; Eun-Cheol Kim; Kim, Hae-Won

    2016-05-01

    Formulating self-setting calcium phosphate cements (CPCs) with secondary phases particularly in the nanoscale order holds great promise to improve biological properties. Here, we focus on the effect that bioactive glass nanoparticles (BGN) incorporated in CPC compositions can have on the proliferation, odontogenic differentiation, and angiogenic stimulation of stem cells derived from human dental pulp (HDPSCs). These odontogenic and angiogenic events are of special importance in the dentin-pulp regeneration processes. In comparison to pure CPCs, nanocomposite cements exhibit a significantly improved proliferation of HDPSCs, and the improvement is more significant as the BGN content increases. The nanocomposite cements substantially enhance the adhesion of cells, and significantly up-regulate odontogenic differentiation, including alkaline phosphatase (ALP) activity and the expressions of odontogenic genes (sialophosphoprotein, dentin matrix protein I, ALP, osteopontin and osteocalcin). Furthermore, the use of nanocomposite cements result in stimulation of angiogenic gene expression (VEGF, FGF-2, VEGFRs, PECAM-1, and VE-cadherin) and protein production (VEGF, VEGFR-1). The angiogenic stimulation by the HDPSCs significantly affects the endothelial cell behaviors, that is, the endothelial cell migration and the tubular network formation are substantially improved when treated with HDPSC-conditioned medium, particularly with the help of nanocomposite cements. The integrin and VEGF signaling pathways are reasoned for the stimulation of the odontogenesis and angiogenesis of cells, where the nanocomposite cements up-regulate the integrin subsets α1, α2, α3, and β1, and activate the integrin downstream signal pathways, such as p-FAK, p-Akt, p-paxillin, JNK, EK, and NF-κB, as well as other nuclear transcriptional factors, including CREB, STAT-3, and ELK-1. The current results indicate that the new formulation of the nanocomposite self-setting cements might provide some

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

    PubMed Central

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

    2015-01-01

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

  4. Isolation of phosphate-solubilizing fungi from phosphate mines and their effect on wheat seedling growth.

    PubMed

    Xiao, Chunqiao; Chi, Ruan; He, Huan; Qiu, Guanzhou; Wang, Dianzuo; Zhang, Wenxue

    2009-11-01

    Three phosphate-solubilizing fungi, identified as Penicillium expansum, Mucor ramosissimus, and Candida krissii, were isolated from phosphate mines (Hubei, People's Republic of China) and characterized. All the isolates demonstrated diverse levels of phosphate-solubilizing capability in National Botanical Research Institute's phosphate growth medium containing rock phosphate as sole phosphate source. Acidification of culture medium seemed to be the main mechanism for rock phosphate solubilization. Indeed, citric acid, oxalic acid, and gluconic acid were shown to be present in the culture medium inoculated with these isolates. Moreover, the isolates produced acid and alkaline phosphatases in culture medium, which may also be helpful for RP solubilization. A strong negative correlation between content of soluble phosphorus and pH (r = - 0.89; p < 0.01) in culture medium was observed in this study. All the isolates promoted growth, soil available phosphorus, phosphorus, and nitrogen uptake of wheat seedling in field soil containing rock phosphate under pot culture conditions, thus demonstrating the capability of these isolates to convert insoluble form of phosphorus into plant available form from rock phosphate, and therefore hold great potential for development as biofertilizers to enhance soil fertility and promote plant growth.

  5. How Climate Change Affects Water Resources in the Alps

    NASA Astrophysics Data System (ADS)

    Schädler, B.

    2009-04-01

    Water resources in the Alps are abundant, but long term observed climatological, glaciological and hydrological time series clearly show ongoing climate changes. And regional climate change scenarios indicate even more changes. Will we experience more severe natural disasters in the Alps and will water scarcity affect alpine agriculture and tourism? Or might the importance of the Alps as «Water Tower of Europe» even grow?

  6. Biochemical characterization of the soluble alkaline phosphatase isolated from the venomous snake W. aegyptia.

    PubMed

    Al-Saleh, Saad S M

    2002-12-01

    A soluble form of alkaline phosphatase (ALP) has been identified and purified from Walterinnesia aegyptia venom using an HPLC system Gold 126/1667 equipped with Protein PAK 125 and Protein PAK 60 columns. The enzyme was purified 3.4 fold over crude venom with a yield of 37.3%. On SDS-PAGE under non-reduced conditions the purified enzyme showed three bands of 212 kD, 80 kD, and 55 kD. However, under reducing conditions, the enzyme showed two bands of 80 kD and 55 kD. The specific activity of ALP was 24 U/mg with p-nitrophenylephosphate as the substrate. During isoelectric focusing experiments the ALP exhibited two bands focused at pH 6.2 and 6.8, which suggests that either the enzyme exists as two different isoforms or the two bands in IEF may be two subunits of 80 kD and 55 kD. The kinetic parameters (Km and Vmax) and IC50 of ALP inhibition by L-phenylalanine, L-leucine, imidazole, caffeine, orthophosphate and permanganate were also investigated in the present study. Zinc and cyanide ions at a concentration of 15 mM and 10 mM, respectively, completely inhibited the activity of W. aegyptia ALP. PMID:12503880

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

    PubMed Central

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

    2016-01-01

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

  8. Possible association between serum alkaline phosphatase concentration and thoracicacute aortic dissection

    PubMed Central

    Yu, Ming; Ding, Juan; Zhao, Long; Huang, Xiang; Ma, Ke-Zhong

    2015-01-01

    Objectives: Alkaline phosphatase (ALP) is an enzyme that catalyzes the hydrolysis of organic pyrophosphate. Accumulating data have demonstrated that the concentration of increased ALP is associated with C-reactive protein (CRP) concentration, and inflammation was complicated in the pathogenesis of acute aortic dissection (ADD). Therefore, the aim of our study was to examine the relationship between serum ALP concentration and thoracic ADD. Methods: We retrieved demographic data and test results of biochemical data of 68 patients with thoracic ADD and 126 Non-thoracic ADD patients, retrospectively. Results: A total of 194 patients were divided into thoracic ADD groups and non-thoracic ADD groups. Age, creatinine(Cr) and high-density lipoprotein cholesterol (HDL-C) were found to be statistical significance between the two groups. The mean ALP level was significantly higher in patients with thoracic ADD compared with Non-thoracic ADD patients (80.6±23.02 Vs. 65.9±16.49, P=0.001). Stepwise multiple logistic regression analyses revealed a significantly association of ALP with thoracic ADD (OR=1.038, 95% CI: 1.015-1.062, P=0.001). In addition, HDL-C was negative associated with thoracic ADD in multiple logistic regression analyses after adjustment for age, sex and Cr (OR=-0.083, 95% CI: 0.012-0.560, P=0.011). Conclusions: The present study suggests that the level of serum ALP is associated with thoracic ADD, and serum ALP concentration may be apotential risk factor for thoracic ADD. PMID:26629214

  9. 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.

  10. 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. PMID:25809455

  11. 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. PMID:19775213

  12. 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.

  13. 13 CFR 120.841 - Qualifications for the ALP.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Development Company Loan Program (504) Accredited Lenders Program (alp) § 120.841 Qualifications for the ALP... staff experience. The CDC's staff must have well-trained, qualified loan officers who are knowledgeable... processing and servicing training. The same loan officer may meet these qualifications. In addition, the...

  14. 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. PMID:26404834

  15. 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.

  16. 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).

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

    PubMed

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

    2013-03-01

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

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

    PubMed

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

    2013-03-01

    Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders. PMID:22743140

  19. Calcium phosphates and glass composite coatings on zirconia for enhanced biocompatibility.

    PubMed

    Kim, Hae-Won; Georgiou, George; Knowles, Jonathan C; Koh, Young-Hag; Kim, Hyoun-Ee

    2004-08-01

    MPa, an improvement of approximately 80% with respect to the pure HA coating. The composite coatings showed much higher dissolution rates than the pure HA coating due to the newly formed crystallines (TCP and DCP) and the remaining glass phase. The osteoblast-like cells grew and spread actively on the composite coating samples. The proliferation numbers and alkaline phosphate (ALP) activities of the cells on the composite coatings were improved by approximately 30-40% when compared to Thermanox control and ZrO(2) substrate, and were comparable to the pure HA coating. These findings suggested that the CaP and P-glass composites are potentially useful for hard tissue coating system, due to their morphological and mechanical integrity, enhanced bioactivity, and favorable responses to the osteoblast-like cells.

  20. 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

  1. 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. PMID:24857506

  2. 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

  3. 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

    Kim, Yongsun; Lee, Seung Hoon; Kang, Byung-Jae; Kim, Wan Hee; Yun, Hui-Suk; Kweon, Oh-Kyeong

    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

  4. Regulation of serum phosphate

    PubMed Central

    Lederer, Eleanor

    2014-01-01

    The regulation of serum phosphate, an acknowledged risk factor for chronic kidney disease and cardiovascular mortality, is poorly understood. The discovery of fibroblast growth factor 23 (FGF23) as a key regulator of renal phosphate handling and activation of vitamin D has revolutionized our comprehension of phosphate homeostasis. Through as yet undetermined mechanisms, circulating and dietary phosphate appear to have a direct effect on FGF23 release by bone cells that, in turn, causes renal phosphate excretion and decreases intestinal phosphate absorption through a decrease in vitamin D production. Thus, the two major phosphaturic hormones, PTH and FGF23, have opposing effects on vitamin D production, placing vitamin D at the nexus of phosphate homeostasis. While our understanding of phosphate homeostasis has advanced, the factors determining regulation of serum phosphate level remain enigmatic. Diet, time of day, season, gender, age and genetics have all been identified as significant contributors to serum phosphate level. The effects of these factors on serum phosphate have major implications for what is understood as ‘normal’ and for studies of phosphate homeostasis and metabolism. Moreover, other hormonal mediators such as dopamine, insulin-like growth factor, and angiotensin II also affect renal handling of phosphate. How the major hormone effects on phosphate handling are regulated and how the effect of these other factors are integrated to yield the measurable serum phosphate are only now beginning to be studied. PMID:24973411

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

    PubMed

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

    2015-06-21

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

  6. Regulation of alkaline phosphatase expression in a neonatal rat clonal calvarial cell strain by retinoic acid.

    PubMed

    Ng, K W; Gummer, P R; Michelangeli, V P; Bateman, J F; Mascara, T; Cole, W G; Martin, T J

    1988-02-01

    A clonal cell strain, UMR 201, was established from a culture of rat calvarial cells by the process of limiting dilution on a collagen substratum. One-day-old neonatal rat calvaria stripped of periosteum were placed on collagen in alpha-MEM with 10% fetal bovine serum (FBS). Cells that grew out from the calvaria were passaged eight times to select cells with the ability to proliferate in culture before cloning was attempted. Cells from the clonal strain were homogeneous in appearance with a doubling time in culture of about 24 hours. The UMR 201 cells formed predominantly type 1 collagen. When treated with retinoic acid (RA), all cells showed an intense staining for alkaline phosphatase (ALP). This effect of RA on the expression of ALP activity was reversible and was time and dose dependent. The earliest change was observed within 6 hours. In contrast, single and isolated clumps of untreated cells stained positively for ALP only when they were confluent. Coincubation with dactinomycin up to 3 hours after the addition of RA completely prevented the expression of ALP, whereas dactinomycin became progressively less effective when added at later times. This is interpreted as indicating a regulatory role of RA on the gene expression of ALP. Other hormones acting on bone, such as 1,25(OH)2 vitamin D3 and dexamethasone, also modulate ALP activity. The cells showed morphologic evidence of senescence after passage 12. Our preliminary studies showed that the UMR 201 cells had the characteristics of relatively undifferentiated mesenchymal cells.(ABSTRACT TRUNCATED AT 250 WORDS)

  7. 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.

  8. Alkaline galvanic cell

    SciTech Connect

    Inoue, T.; Maeda, Y.; Momose, K.; Wakahata, T.

    1983-10-04

    An alkaline galvanic cell is disclosed including a container serving for a cathode terminal, a sealing plate in the form of a layered clad plate serving for an anode terminal to be fitted into the container, and an insulating packing provided between the sealing plate and container for sealing the cell upon assembly. The cell is provided with a layer of epoxy adduct polyamide amine having amine valence in the range of 50 to 400 and disposed between the innermost copper layer of the sealing plate arranged to be readily amalgamated and the insulating packing so as to serve as a sealing agent or liquid leakage suppression agent.

  9. Culture human mesenchymal stem cells with calcium phosphate cement scaffolds for bone repair.

    PubMed

    Weir, Michael D; Xu, Hockin H K

    2010-04-01

    Because of its moldability and excellent osteoconductivity, calcium phosphate cement (CPC) is highly promising for craniofacial and orthopedic applications. The objectives of this study were to investigate the response of human mesenchymal stem cells (hMSCs) to a high-strength CPC-chitosan scaffold and to examine cell proliferation and osteogenic differentiation. hMSCs were seeded onto CPC-chitosan composite, CPC control, and tissue culture polystyrene (TCPS). Alkaline phosphatase activity (ALP) and mineralization of hMSCs were measured. CPC-chitosan had a flexural strength (mean + or - SD; n = 5) of (19.5 + or - 1.4) MPa, higher than (8.0 + or - 1.4) MPa of CPC control (p < 0.05). The percentage of live hMSCs on CPC-chitosan was (90.5 + or - 1.3)% at 8 days, matching (90.7 + or - 3.8)% of CPC control (p > 0.1). The CPC-chitosan surface area covered by the attached hMSCs increased from (51 + or - 11)% at 1 day to (90 + or - 4)% at 8 days (p < 0.05), matching those of CPC control (p > 0.1). Hence, the CPC strength was significantly increased via chitosan without compromising the hMSC response. At 8 days, there was a significant increase in ALP of cells in osteogenic media (10.99 + or - 0.93) [(mM pNpp/min)/(microg DNA)] versus control media (3.62 + or - 0.40) (p < 0.05). hMSCs in osteogenic media exhibited greater mineralization area of (47.5 + or - 19.7)% compared with (6.1 + or - 2.3)% in control medium on TCPS (p < 0.05). In conclusion, hMSCs showed excellent attachment and viability on the strong and tough CPC-chitosan scaffold, matching the hMSC response on CPC control. hMSCs were successfully differentiated down the osteogenic lineage. Hence, the strong, in situ hardening CPC-chitosan scaffold may be useful as a moderate load-bearing vehicle to deliver hMSCs for maxillofacial and orthopedic bone tissue engineering. PMID:20091907

  10. 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

  11. Role of alkaline phosphatase from Manduca sexta in the mechanism of action of Bacillus thuringiensis Cry1Ab toxin.

    PubMed

    Arenas, Iván; Bravo, Alejandra; Soberón, Mario; Gómez, Isabel

    2010-04-23

    Cry toxins produced by Bacillus thuringiensis have been recognized as pore-forming toxins whose primary action is to lyse midgut epithelial cells in their target insect. In the case of the Cry1A toxins, a prepore oligomeric intermediate is formed after interaction with cadherin receptor. The Cry1A oligomer then interacts with glycosylphosphatidylinositol-anchored receptors. Two Manduca sexta glycosylphosphatidylinositol-anchored proteins, aminopeptidase (APN) and alkaline phosphatase (ALP), have been shown to bind Cry1Ab, although their role in toxicity remains to be determined. Detection of Cry1Ab binding proteins by ligand blot assay revealed that ALP is preferentially expressed earlier during insect development, because it was found in the first larval instars, whereas APN is induced later after the third larval instar. The binding of Cry1Ab oligomer to pure preparations of APN and ALP showed that this toxin structure interacts with both receptors with high affinity (apparent K(d) = 0.6 nM), whereas the monomer showed weaker binding (apparent K(d) = 101.6 and 267.3 nM for APN and ALP, respectively). Several Cry1Ab nontoxic mutants located in the exposed loop 2 of domain II or in beta-16 of domain III were affected in binding to APN and ALP, depending on their oligomeric state. In particular monomers of the nontoxic domain III, the L511A mutant did not bind ALP but retained APN binding, suggesting that initial interaction with ALP is critical for toxicity. Our data suggest that APN and ALP fulfill two roles. First APN and ALP are initial receptors promoting the localization of toxin monomers in the midgut microvilli before interaction with cadherin. Then APN and ALP function as secondary receptors mediating oligomer insertion into the membrane. However, the expression pattern of these receptors and the phenotype of L511A mutant suggest that ALP may have a predominant role in toxin action because Cry toxins are highly effective against the neonate larvae that is

  12. Alkaline fuel cells applications

    NASA Astrophysics Data System (ADS)

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

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

  13. 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.

  14. 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.

  15. 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

  16. Automated Laser Paint Stripping (ALPS) update

    NASA Astrophysics Data System (ADS)

    Lovoi, Paul

    1993-03-01

    To date, the DoD has played a major role in funding a number of paint stripping programs. Some technologies have proven less effective than contemplated. Others are still in the validation phase. Paint stripping is one of the hottest issues being addressed by the finishing industry since the Environmental Protection Agency (EPA) has mandated that chemical stripping using methylene chloride/phenolic type strippers be stopped. The DoD and commercial aircraft companies are hard-pressed to find an alternative. Automated laser paint stripping has been identified as a technique for removing coatings from aircraft surfaces. International Technical Associates (InTA) was awarded a Navy contract for an automated laser paint stripping system (ALPS) that will remove paint from metallic and composite substrates. For the program, which will validate laser paint stripping, InTA will design, build, test, and install a system for fighter-sized aircraft at both the Norfolk and North Island (San Diego) Aviation Depots.

  17. Enhancement of osteoblastic differentiation in alginate gel beads with bioactive octacalcium phosphate particles.

    PubMed

    Endo, Kosei; Anada, Takahisa; Yamada, Masumi; Seki, Minoru; Sasaki, Keiichi; Suzuki, Osamu

    2015-12-01

    The present study investigated whether alginate (Alg) hydrogel microbeads have a role in maintaining mouse bone marrow stromal ST-2 cells and release the cells after being stimulated by synthetic octacalcium phosphate (OCP), which is a mineral crystal capable of stimulating osteoblastic differentiation during a conversion process to hydroxyapatite (HA). The ST-2 cell suspension in the alginate solution, which contained various concentrations of OCP granules with diameters less than 53 μm, was extruded drop-wise into a stirred gelation solution containing BaCl2 using an encapsulator with nitrogen gas stream. The Alg-microbeads (Alg/OCP · ST-2 microbeads) that were generated, which had a diameter of approximately 400 μm, were incubated for up to 14 d and then assessed for osteoblastic differentiation. Alg-microbeads with cells were also incubated to identify the possible conversion from OCP to HA. Osteoblast differentiation markers in ST-2 cells, alkaline phosphatase (ALP) and collagen type I, were up-regulated in the presence of higher amounts of OCP. X-ray diffraction analysis and Fourier transform infrared spectroscopy confirmed that the OCP tended to convert to HA over time, suggesting that the OCP in Alg-microbeads interacts three-dimensionally with ST-2 cells and stimulates its osteoblastic differentiation. The release of ST-2 cells from the microbeads was also estimated. ST-2 cells were identified outside of the microbeads, although the cell number tended to decrease with increasing OCP. These results suggest that Alg/OCP microbeads could be used as a vehicle to activate osteoblastic cells and deliver them to sites where bone regeneration is needed. PMID:26657659

  18. Discovery of Alkaline Volcanic Rocks on Mars

    NASA Astrophysics Data System (ADS)

    McSween, H. Y.; Team, A. S.

    2006-05-01

    Based on remote sensing measurements and the compositions of martian meteorites, the surface of Mars is inferred to be dominated by subalkaline mafic volcanic rocks. However, the Spirit rover has recently discovered lavas of alkalic composition. Picritic (Adirondack class) basalts with high alkali and low silica contents were previously analyzed on the plains of Gusev Crater, and two new classes of dark, fine-grained, relatively unaltered volcanic rocks with distinctive thermal emission spectra have now been found as float and in a possible dike at high elevations in the Columbia Hills. Chemical analyses indicate that these rocks are mildly alkaline basalt and trachybasalt, 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. Chemical compositions of these rocks lie along a MELTS-calculated liquid line of descent for Adirondack class basalt. Systematic changes in normative mineralogy are consistent with the calculated magmatic fractionation. We infer that Backstay- and Irvine-class magmas may have formed by low-pressure fractionation of primitive, oxidized Adirondack-class magmas and were possibly emplaced coevally with the plains basalts. The compositions of 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. This discovery may have implications for the composition of the martian mantle source region and the conditions under which it melted.

  19. Molecular modeling of human alkaline sphingomyelinase.

    PubMed

    Suresh, Panneer Selvam; Olubiyi, Olujide; Thirunavukkarasu, Chinnasamy; Strodel, Birgit; Kumar, Muthuvel Suresh

    2011-01-01

    Alkaline sphingomyelinase, which is expressed in the human intestine and hydrolyses sphingomyelin, is a component of the plasma and the lysosomal membranes. Hydrolase of sphingomyelin generates ceramide, sphingosine, and sphingosine 1-phosphate that have regulatory effects on vital cellular functions such as proliferation, differentiation, and apoptosis. The enzyme belongs to the Nucleotide Pyrophosphatase/Phosphodiesterase family and it differs in structural similarity with acidic and neutral sphingomyelinase. In the present study we modeled alkaline sphingomyelinase using homology modeling based on the structure of Nucleotide Pyrophosphatase/Phosphodiesterase from Xanthomonas axonopodis with which it shares 34% identity. Homology modeling was performed using Modeller9v7. We found that Cys78 and Cys394 form a disulphide bond. Further analysis shows that Ser76 may be important for the function of this enzyme, which is supported by the findings of Wu et al. (2005), that S76F abolishes the activity completely. We found that the residues bound to Zn(2+) are conserved and geometrically similar with the template. Molecular Dynamics simulations were carried out for the modeled protein to observe the effect of Zinc metal ions. It was observed that the metal ion has little effect with regard to the stability but induces increased fluctuations in the protein. These analyses showed that Zinc ions play an important role in stabilizing the secondary structure and in maintaining the compactness of the active site. PMID:21544170

  20. 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.

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

    PubMed

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

    2016-06-01

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

  2. Pyrophosphate-regulated Zn(2+)-dependent DNAzyme activity: an amplified fluorescence sensing strategy for alkaline phosphatase.

    PubMed

    Kong, Rong-Mei; Fu, Ting; Sun, Ni-Na; Qu, Feng-Li; Zhang, Shu-Fang; Zhang, Xiao-Bing

    2013-12-15

    In this work, based on the fact that pyrophosphate (PPi) could regulate the activity of Zn(2+)-dependent DNAzyme, we for the first time report a fluorescence turn-on sensing system for alkaline phosphatase (ALP) with improved sensitivity via nonprotein-enzymatic signal amplification. A catalytic and molecular beacon (CAMB) design was employed to further improve its sensitivity. Taking advantage of the strong interactions between PPi and the Zn(2+), the cofactor Zn(2+) was caged, and the DNAzyme activity was effectively inhibited. The introduction of ALP, however, could catalyze the hydrolysis of PPi and release free Zn(2+), resulting in the activation of DNAzyme to catalyze the cleavage of the molecular beacon substrate with a remarkable increase of fluorescent signal. These optimized designs together allow a high sensitivity for ALP, with a detection limit of 20 pM observed, much lower than previously reported methods. It has also been used for detection of ALP in human serum with satisfactory results, demonstrating its potential applications in clinical diagnosis.

  3. Glucose-6-phosphate dehydrogenase

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003671.htm Glucose-6-phosphate dehydrogenase test To use the sharing features on this page, please enable JavaScript. Glucose-6-phosphate dehydrogenase (G6PD) is a type of ...

  4. Uranium from phosphate ores

    SciTech Connect

    Hurst, F.J.

    1983-01-01

    The following topics are described briefly: the way phosphate fertilizers are made; how uranium is recovered in the phosphate industry; and how to detect covert uranium recovery operations in a phsophate plant.

  5. Reliable Digital Single Molecule Electrochemistry for Ultrasensitive Alkaline Phosphatase Detection.

    PubMed

    Wu, Zhen; Zhou, Chuan-Hua; Pan, Liang-Jun; Zeng, Tao; Zhu, Lian; Pang, Dai-Wen; Zhang, Zhi-Ling

    2016-09-20

    Single molecule electrochemistry (SME) has gained much progress in fundamental studies, but it is difficult to use in practice due to its less reliability. We have solved the reliability of single molecule electrochemical detection by integration of digital analysis with efficient signal amplification of enzyme-induced metallization (EIM) together with high-throughput parallelism of microelectrode array (MA), establishing a digital single molecule electrochemical detection method (dSMED). Our dSMED has been successfully used for alkaline phosphatase (ALP) detection in the complex sample of liver cancer cells. Compared to direct measurement of the oxidation current of enzyme products, EIM can enhance signals by about 100 times, achieving signal-to-background ratio high enough for single molecule detection. The integration of digital analysis with SME can further decrease the detection limit of ALP to 1 aM relative to original 50 aM, enabling dSMED to be sensitively, specifically and reliably applied in liver cancer cells. The presented dSMED is enormously promising in exploring physical and chemical properties of single molecules, single biomolecular detection, or single-cell analysis.

  6. Silica in alkaline brines

    USGS Publications Warehouse

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

    1967-01-01

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

  7. Bifunctional alkaline oxygen electrodes

    NASA Technical Reports Server (NTRS)

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

    1991-01-01

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

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

    PubMed

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

    1993-10-15

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

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

    PubMed

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

    2006-02-15

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

  10. Regulation of Phosphate Accumulation in the Unicellular Cyanobacterium Synechococcus

    PubMed Central

    Grillo, John F.; Gibson, Jane

    1979-01-01

    The phosphorus contents of acid-soluble pools, lipid, ribonucleic acid, and acid-insoluble polyphosphate were lowered in Synechococcus in proportion to the reduction in growth rate in phosphate-limited but not in nitrate-limited continuous culture. Phosphorus in these cell fractions was lost proportionately during progressive phosphate starvation of batch cultures. Acid-insoluble polyphosphate was always present in all cultural conditions to about 10% of total cell phosphorus and did not turn over during balanced exponential growth. Extensive polyphosphate formation occurred transiently when phosphate was given to cells which had been phosphate limited. This material was broken down after 8 h even in the presence of excess external orthophosphate, and its phosphorus was transferred into other cell fractions, notably ribonucleic acid. Phosphate uptake kinetics indicated an invariant apparent Km of about 0.5 μM, but Vmax was 40 to 50 times greater in cells from phosphate-limited cultures than in cells from nitrate-limited or balanced batch cultures. Over 90% of the phosphate taken up within the first 30 s at 15°C was recovered as orthophosphate. The uptake process is highly specific, since neither phosphate entry nor growth was affected by a 100-fold excess of arsenate. The activity of polyphosphate synthetase in cell extracts increased at least 20-fold during phosphate starvation or in phosphate-restricted growth, but polyphosphatase activity was little changed by different growth conditions. The findings suggest that derepression of the phosphate transport and polyphosphate-synthesizing systems as well as alkaline phosphatase occurs in phosphate shortage, but that the breakdown of polyphosphate in this organism is regulated by modulation of existing enzyme activity. PMID:227842

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

    PubMed

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

    2015-10-20

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

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

    PubMed

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

    2015-10-20

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

  13. Effect of cobalt on synthesis and activation of Bacillus licheniformis alkaline phosphatase.

    PubMed Central

    Spencer, D B; Chen, C P; Hulett, F M

    1981-01-01

    The effect of CO2+ on the synthesis and activation of Bacillus licheniformis MC14 alkaline phosphatase has been shown by the development of a defined minimal salts medium in which this organism produces 35 times more (assayable) alkaline phosphatase than when grown in a low-phosphate complex medium or in the defined medium without cobalt. Stimulation of enzyme activity with cobalt is dependent on a low phosphate concentration in the medium (below 0.075 mM) and continued protein synthesis. Cobalt stimulation resulted in alkaline phosphate production being a major portion of total protein synthesized during late-logarithmic and early-stationary-phase culture growth. Cells cultured in the defined medium minus cobalt, or purified enzyme partially inactivated with a chelating agent, showed a 2.5-fold increase in activity when assayed in the presence of cobalt. Atomic spectral analysis indicated the presence of 3.65 +/- 0.45 g-atoms of cobalt associated with each mole of purified active alkaline phosphatase. A biochemical localization as a function of culture age in this medium showed that alkaline phosphatase was associated with the cytoplasmic membrane and was also found as a soluble enzyme in the periplasmic region and secreted into the growth medium. PMID:7462163

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

    PubMed

    Sibirnyi, A A; Shavlovskii, G M

    1978-01-01

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

  15. Osteopontin involvement in integrin-mediated cell signaling and regulation of expression of alkaline phosphatase during early differentiation of UMR cells.

    PubMed

    Liu, Y K; Uemura, T; Nemoto, A; Yabe, T; Fujii, N; Ushida, T; Tateishi, T

    1997-12-22

    To clarify the function of osteopontin in osteoblast differentiation, we have examined the signal transduction pathway in an osteoblastic cell line (UMR106-6) bound to osteopontin, fibronectin, vitronectin and collagen type I surfaces. This was done by investigating the production and autophosphorylation of focal adhesion kinase (FAK) and the expression of alkaline phosphatase (ALP) at the transcription level. Results suggest that osteopontin was not only responsible for the autophosphorylation of FAK but regulated the expression of ALP, which was strongly correlated with FAK activity. These results suggest that osteopontin might act as a trigger in the early differentiation of osteoblasts.

  16. 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

  17. [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.

  18. 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

  19. 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.

  20. Activity of soil dehydrogenases, urease, and acid and alkaline phosphatases in soil polluted with petroleum.

    PubMed

    Wyszkowska, Jadwiga; Wyszkowski, Mirosław

    2010-01-01

    This study was undertaken to (1) determine the effects of petroleum pollution on changes in the biochemical properties of soil and (2) demonstrate whether the application of compost, bentonite, and calcium oxide is likely to restore biological balance. Petroleum soil pollution at a dose ranging from 2.5 to 10 cm(3)/kg disturbed the biochemical balance as evidenced by inhibition of the activities of soil dehydrogenases (SDH), urease (URE), and acid phosphatase (ACP). The greatest change was noted in the activity of SDH, whereas the least change occurred in URE. Petroleum significantly increased the activity of soil alkaline phosphatase (ALP) in soil used for spring rape, whereas in soil used for oat harvest there was decreased ALP activity. The application of compost, bentonite, and calcium oxide to soil proved effective in mitigating the adverse effects of petroleum on the activities of soil enzymes. Soil enrichment with compost, bentonite, and calcium oxide was found to stimulate the activities of URE and ALP and inhibit the activity of ACP. The influence of bentonite and calcium oxide was greater than that of compost. Calcium oxide and, to a lesser extent, compost were found to increase the activity of SDH, whereas bentonite exerted the opposite effect, especially in the case of the main crop, spring rape. The activities of SDH, URE, and ACP were higher in soil used for rape than that for oats. In contrast the activity of ALP was higher in soil used for oats. Data thus indicate that compost and especially bentonite and calcium oxide exerted a positive effect on activities of some enzymes in soil polluted with petroleum. Application of neutralizing additives to soil restored soil biological balance by counteracting the negative influence of petroleum on activities of URE and ALP. PMID:20706945

  1. 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. PMID:27612703

  2. Evaluation of Alkaline Cleaner Materials

    NASA Technical Reports Server (NTRS)

    Partz, Earl

    1998-01-01

    Alkaline cleaners used to process aluminum substrates have contained chromium as the corrosion inhibitor. Chromium is a hazardous substance whose use and control are described by environmental laws. Replacement materials that have the characteristics of chromated alkaline cleaners need to be found that address both the cleaning requirements and environmental impacts. This report will review environmentally friendly candidates evaluated as non-chromium alkaline cleaner replacements and methods used to compare those candidates one versus another. The report will also list characteristics used to select candidates based on their declared contents. It will also describe and evaluate methods used to discriminate among the large number of prospective candidates.

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

    PubMed Central

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

    2015-01-01

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

  4. 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. PMID:25875245

  5. 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

  6. 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.

  7. 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

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

    PubMed

    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

  9. Differential Role of Manduca sexta Aminopeptidase-N and Alkaline Phosphatase in the Mode of Action of Cry1Aa, Cry1Ab, and Cry1Ac Toxins from Bacillus thuringiensis

    PubMed Central

    Flores-Escobar, Biviana; Rodríguez-Magadan, Hector; Bravo, Alejandra; Soberón, Mario

    2013-01-01

    Aminopeptidase-N (APN1) and alkaline phosphatase (ALP) proteins located in the midgut epithelium of Manduca sexta have been implicated as receptors for Cry1Aa, Cry1Ab, and Cry1Ac insecticidal proteins produced by Bacillus thuringiensis subsp. kurstaki. In this study, we analyzed the roles of ALP and APN1 in the toxicity of these three Cry1A proteins. Ligand blot analysis using brush border membrane vesicles of M. sexta showed that Cry1Aa and Cry1Ab bind preferentially to ALP during early instars while binding to APN was observed after the third instar of larval development. Cry1Ac binds to APN throughout all larval development, with no apparent binding to ALP. ALP was cloned from M. sexta midgut RNA and expressed in Escherichia coli. Surface plasmon resonance binding analysis showed that recombinant ALP binds to Cry1Ac with 16-fold lower affinity than to Cry1Aa or Cry1Ab. Downregulation of APN1 and ALP expression by RNA interference (RNAi) using specific double-stranded RNA correlated with a reduction of transcript and protein levels. Toxicity analysis of the three Cry1A proteins in ALP- or APN1-silenced larvae showed that Cry1Aa relies similarly on both receptor molecules for toxicity. In contrast, RNAi experiments showed that ALP is more important than APN for Cry1Ab toxicity, while Cry1Ac relied principally on APN1. These results indicated that ALP and APN1 have a differential role in the mode of action of Cry1A toxins, suggesting that B. thuringiensis subsp. kurstaki produces different Cry1A toxins that in conjunction target diverse midgut proteins to exert their insecticidal effect. PMID:23686267

  10. Water Management Strategies against Water Shortage in the Alps (Invited)

    NASA Astrophysics Data System (ADS)

    de Jong, C.

    2009-12-01

    In the European Alps water has been perceived as ubiquitous and not the subject of management. Climate change and anthropogenic pressures have changed demand and supply relations rapidly and over the last 10 years, water problems have increasingly become apparent over temporal and spatial hotspots. Stakeholders in the Alpine Space have been confronted with water management problems in agriculture, tourism and hydropower to such an extent that they approached scientists to create solution strategies based on adaptation and mitigation. In this context, Alp-Water-Scarce, a European project on Water Management Strategies against Water Scarcity in the Alps was funded by the Alpine Space programme as part of the "European Territorial Cooperation" scheme. It has 17 project partners from Austria Switzerland, France, Italy and Slovenia from local governments, provinces, federal institutes and offices, universities, regional agencies, alpine societies, geological surveys, and chambers of agriculture and forestry. The Lead Partner is the Mountain Institute in Savoy, Rhone-Alpes, France. The main challenges of this project are to create local Early Warning Systems against Water Scarcity in the Alps. This system is based on strengthening existing long-term monitoring and modeling and creating new measuring networks in those countries where they do not yet exist. It is anchored strongly and actively within a Stakeholder Interaction Forum linked across comparative and contrasting regions across the Alps. The Early Warning System is based on the linkage and improvement of field monitoring and assemblage of qualitative and quantitative data derived both from natural water reservoirs as well as from anthropogenic water use in 28 selected pilot regions selected in France, Italy, Austria, Slovenia and Switzerland. The objectives are to improve water management at the short term (annual scale) and long term (using future scenarios) based on modelling and application of climate change

  11. 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

    2007-05-01

    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).

  12. 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

    2007-05-01

    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). PMID:17030413

  13. Ultrasensitive electroanalysis of low-level free microRNAs in blood by maximum signal amplification of catalytic silver deposition using alkaline phosphatase-incorporated gold nanoclusters.

    PubMed

    Si, Yanmei; Sun, Zongzhao; Zhang, Ning; Qi, Wei; Li, Shuying; Chen, Lijun; Wang, Hua

    2014-10-21

    An ultrasensitive sandwich-type analysis method has been initially developed for probing low-level free microRNAs (miRNAs) in blood by a maximal signal amplification protocol of catalytic silver deposition. Gold nanoclusters (AuNCs) were first synthesized and in-site incorporated into alkaline phosphatase (ALP) to form the ALP-AuNCs. Unexpectedly, the so incorporated AuNCs could dramatically enhance the catalysis activities of ALP-AuNCs versus native ALP. A sandwiched hybridization protocol was then proposed using ALP-AuNCs as the catalytic labels of the DNA detection probes for targeting miRNAs that were magnetically caught from blood samples by DNA capture probes, followed by the catalytic ligation of two DNA probes complementary to the targets. Herein, the ALP-AuNC labels could act as the bicatalysts separately in the ALP-catalyzed substrate dephosphorylation reaction and the AuNCs-accelerated silver deposition reaction. The signal amplification of ALP-AuNCs-catalyzed silver deposition was thereby maximized to be measured by the electrochemical outputs. The developed electroanalysis strategy could allow for the ultrasensitive detection of free miRNAs in blood with the detection limit as low as 21.5 aM, including the accurate identification of single-base mutant levels in miRNAs. Such a sandwich-type analysis method may circumvent the bottlenecks of the current detection techniques in probing short-chain miRNAs. It would be tailored as an ultrasensitive detection candidate for low-level free miRNAs in blood toward the diagnosis of cancer and the warning or monitoring of cancer metastasis in the clinical laboratory.

  14. 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

  15. 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.

  16. 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.

  17. Phosphorus, phosphorous, and phosphate.

    PubMed

    Iheagwara, O Susan; Ing, Todd S; Kjellstrand, Carl M; Lew, Susie Q

    2013-10-01

    This article distinguishes the terms "phosphorus, phosphorous, and phosphate" which are frequently used interchangeably. We point out the difference between phosphorus and phosphate, with an emphasis on the unit of measure. Expressing a value without the proper name or unit of measure may lead to misunderstanding and erroneous conclusions. We indicate why phosphate must be expressed as milligrams per deciliter or millimoles per liter and not as milliequivalents per liter. Therefore, we elucidate the distinction among the terms "phosphorus, phosphorous, and phosphate" and the importance of saying precisely what one really means.

  18. Isolation of 3-phosphohistidine from phosphorylated pyruvate, phosphate dikinase.

    PubMed Central

    Spronk, A M; Yoshida, H; Wood, H G

    1976-01-01

    Pyruvate, phosphate dikinase (EC 2-7-9-1) catalyzes formation of phosphoenolpyruvate, AMP, and inorganic pyrophosphate from pyruvate, ATP, and orthophosphate. A pyrophosphoryl and phosphoryl form of the enzyme is involved in this transfer. The [32P]phosphoryl form of pyruvate, phosphate dikinase was prepared with enzyme isolated from Bacteroides symbiosus. The [32P]phosphoryl enzyme was found to have properties corresponding to a phosphoramidate linkage and this was confirmed by isolation of 3-[32P]phosphohistidine from alkaline hydrolysates of the enzyme. The histidyl residue is considered to be the pyrophosphoryl- and phosphoryl-carrier between the three substrate sites of this enzyme. PMID:12506

  19. 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

  20. Lactobacillus rhamnosus strain GG restores alkaline phosphatase activity in differentiating Caco-2 cells dosed with the potent mycotoxin deoxynivalenol.

    PubMed

    Turner, P C; Wu, Q K; Piekkola, S; Gratz, S; Mykkänen, H; El-Nezami, H

    2008-06-01

    Deoxynivalenol (DON) contamination of cereal crops occurs frequently, and may cause acute exposure at high levels or chronic more moderate exposure. DON has proven toxicity including restriction of enterocyte differentiation, which may play a part in DON induced gastroenteritis. The probiotic bacteria Lactobacillus rhamnosus strain GG (GG) can bind DON, and therefore potentially restrict bioavailability of this toxin. Binding efficacy is not significantly altered by heat treatment, and therefore this in vitro study evaluated whether heat inactivated GG could restore the differentiation process in Caco-2 cells, using alkaline phosphatase (ALP) activity as a marker of differentiation. DON (200ng/mL) caused a significant (p<0.001) 36% reduction in ALP activity (1598+/-137U/mg protein) compared to untreated cells (2502+/-80U/mg). A dose dependant restoration of ALP activity was observed where DON treated cells were co-incubated with heat inactivated GG (1719+/-84; 2007+/-142; 2272+/-160U/mg for GG at 1x10(4) (p>0.9), 1x10(7) (p<0.001), and 1x10(10)CFU/mL (p<0.001), respectively). Co-incubation of the non-binding strain, LC-705 (1x10(10)CFU/mL), with DON did not significantly restore the ALP (1841+/-97U/mg, p<0.077) compared to DON only treated cells. When viable GG were co-incubated with DON a similar restoration of ALP activity was observed as seen for heat inactivated GG. These combined data suggest that the major effect of GG on restoring ALP activity, and therefore Caco-2 cell differentiation, was due to specific binding of DON, with possibly a more minor role of non-specific bacterial interference.

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. 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…

  6. 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

  7. 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.

  8. Present-day uplift of the western Alps

    NASA Astrophysics Data System (ADS)

    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.

    2016-06-01

    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.

  9. Crustal structure and active tectonics in the Eastern Alps

    NASA Astrophysics Data System (ADS)

    Brückl, E.; Behm, M.; Decker, K.; Grad, M.; Guterch, A.; Keller, G. R.; Thybo, H.

    2010-04-01

    During the last decade, a series of controlled source seismic experiments brought new insight into the crustal and lithospheric structure of the Eastern Alps and their adjacent tectonic provinces. A fragmentation of the lithosphere into three blocks, Europe (EU), Adria (AD), and the new Pannonian fragment (PA), was interpreted and a triple junction was inferred. The goal of this study has been to relate these deep crustal structures to active tectonics. We used elastic plate modeling to reconsider the Moho fragmentation. We interpret subduction of EU below AD and PA from north to south and underthusting of AD mantle below PA from southwest to northeast. The Moho fragmentation correlates well with major upper crustal structures and is supported by gravity, seismic, and geodetic data. An analysis of crustal thickening suggests that active convergence is associated with continued thrusting and lateral extrusion in the central Eastern Alps and thickening of the Adriatic indenter under the Southern Alps. According to the velocity relations at the triple junction, PA moves relative to EU and AD along ENE and SE striking faults, mainly by strike slip. An eastward directed extensional component is compensated by the lateral extrusion of the central Eastern Alps. The Periadriatic (Insubric) line east of the triple junction and the mid-Hungarian fault zone have relatively recently lost their role as first-order active structures. We favor the idea that the Pannonian fragment and the TISZA block merged to a "soft" microplate surrounded by the Eastern and Southern Alpine, Carpathian, and Dinaric orogens.

  10. 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.

  11. 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)

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

    USGS Publications Warehouse

    Nagle, David B.; Wright, C. Wayne

    2016-01-01

    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.

  13. 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. PMID:15756826

  14. 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

  15. 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.

  16. 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)

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

    PubMed

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

    2015-01-01

    Development of an efficient expression system for (especially secreted) bacterial lytic enzymes is a complicated task due to the specificity of their action. The substrate for such enzymes is peptidoglycan, the main structural component of bacterial cell walls. For this reason, expression of recombinant lytic proteins is often accompanied with lysis of the producing bacterium. This paper presents data on the construction of an inducible system for expression of the lytic peptidases AlpA and AlpB from Lysobacter sp. XL1 in Pseudomonas fluorescens Q2-87, which provides for the successful secretion of these proteins into the culture liquid. In this system, the endopeptidase gene under control of the T7lac promoter was integrated into the bacterial chromosome, as well as the Escherichia coli lactose operon repressor protein gene. The T7 pol gene under lac promoter control, which encodes the phage T7 RNA polymerase, is maintained in Pseudomonas cells on the plasmids. Media and cultivation conditions for the recombinant strains were selected to enable the production of AlpA and AlpB by a simple purification protocol. Production of recombinant lytic enzymes should contribute to the development of new-generation antimicrobial drugs whose application will not be accompanied by selection of resistant microorganisms. PMID:26138026

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

    PubMed

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

    2015-01-01

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

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

    PubMed

    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.

  20. 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.

  1. 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

  2. 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.

  3. 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

  4. Alkaline Phosphatase-Mimicking Peptide Nanofibers for Osteogenic Differentiation.

    PubMed

    Gulseren, Gulcihan; Yasa, I Ceren; Ustahuseyin, Oya; Tekin, E Deniz; Tekinay, Ayse B; Guler, Mustafa O

    2015-07-13

    Recognition of molecules and regulation of extracellular matrix synthesis are some of the functions of enzymes in addition to their catalytic activity. While a diverse array of enzyme-like materials have been developed, these efforts have largely been confined to the imitation of the chemical structure and catalytic activity of the enzymes, and it is unclear whether enzyme-mimetic molecules can also be used to replicate the matrix-regulatory roles ordinarily performed by natural enzymes. Self-assembled peptide nanofibers can provide multifunctional enzyme-mimetic properties, as the active sequences of the target enzymes can be directly incorporated into the peptides. Here, we report enhanced bone regeneration efficiency through peptide nanofibers carrying both catalytic and matrix-regulatory functions of alkaline phosphatase, a versatile enzyme that plays a critical role in bone formation by regulating phosphate homeostasis and calcifiable bone matrix formation. Histidine presenting peptide nanostructures were developed to function as phosphatases. These molecules are able to catalyze phosphate hydrolysis and serve as bone-like nodule inducing scaffolds. Alkaline phosphatase-like peptide nanofibers enabled osteogenesis for both osteoblast-like and mesenchymal cell lines.

  5. Jura, Alps and the boundary of the Adria subplate

    NASA Astrophysics Data System (ADS)

    Laubscher, Hans

    2010-03-01

    The thin-skinned Jura fold/thrust belt is the frontal part of a thick-skinned late Miocene Alpine nappe. Important late Miocene structures in the Alps that help define this nappe are the External Massifs in the north, the Orobic thrusts and the Piemontese Southern Alps in the south, the Giudicarie-Brenner Line in the east and, more vaguely, the south end of the Belledonne Massif in the west. A bottom boundary compatible with the data is the Brittle-Ductile Transition (BDT). This Jura nappe is separated from the (latest Oligocene)-early Miocene Helvetic nappe system most obviously by the late Miocene Orobic-Giudicarie-Brenner fault complex. In contrast, the distinctive boundary of the early Miocene thick-skinned Helvetic nappe was the Iorio-Tonale Line (IT) in the south, merging into the Piemontese Southern Alps at what may be termed the "Locarno Singularity"-the point of minimum width of the thick-skinned Alps; the northern boundary passed through the Gotthard-and Montblanc Massifs. Motion along the southern boundary was dextrally transpressive, associated with complex strain partitioning patterns. One part of these patterns is the confusing entity comprising the Piemontese Southern Alps and the Lepontine "dome". The slivers composing the Piemontese Southern Alps are dragged dextrally by the Iorio-Tonale strike-slip at the "Locarno Singularity" and thereby exhibit an important characteristic trait of "exotic terranes" — crustal slivers dragged along strike-slip faults. The Lepontine "dome" around the Locarno Singularity on the north side of the Iorio-Tonale Line displays the characteristics of a backstop in thrust systems as modeled in sandbox experiments. The special boundary functioning as a backstop, compatible with the transpressive kinematics, was the Ivrea body, a strong mafic-ultramafic-granulitic complex which in the early Miocene migrated dextrally along the Iorio-Tonale Line, forcing detachment to descend into the hot middle crust and producing an

  6. 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.

  7. Genotype-by-sex and environment-by-sex interactions influence variation in serum levels of bone-specific alkaline phosphatase in adult baboons (Papio hamadryas).

    PubMed

    Havill, L M; Mahaney, M C; Rogers, J

    2004-07-01

    While more than 77% of the people in the US with osteoporosis are women, the contributions of genotype-by-sex (G x S) and environment-by-sex interactions to sex differences in osteoporosis risk factors have not been studied. To address this issue, we conducted a statistical genetic analysis of serum concentrations of bone-specific alkaline phosphatase (Bone ALP), a highly specific marker of osteoblast function that is elevated in persons with conditions like osteoporosis characterized by excessive bone turnover or rapid bone loss. We assayed Bone ALP from 657 pedigreed baboons using a commercially available ELISA kit. Using a maximum likelihood variance decomposition approach, we treated sex as an environmental milieu in which genes influencing Bone ALP levels are expressed. We modeled the genetic covariance in Bone ALP between all relative pairs conditional on their sex so that the covariance is the product of the kinship, the genetic correlation between trait levels in the two sexes, and the genetic variances in the two sexes. Sex-specific maximum likelihood estimates (MLE) of residual heritability for Bone ALP were greater for females than for males (h2 = 0.44 vs. h2 = 0.26, respectively), but likelihood ratio tests revealed only a marginally significant difference in sex-specific genetic variances (P = 0.057). In contrast, the between-sex genetic correlation (rhoG = 0.43) was significantly less than 1.0 (P = 0.037), and the difference in sex-specific environmental variances was highly significant (P = 0.00006). We report the first evidence for G x S interactions influencing variation in an osteoporosis risk factor. The diminished between-sex genetic correlation implies that different genes influence Bone ALP levels in the two sexes. The significant differences between environmental variances suggest that unmeasured factors, including those from the internal, biological environments of the two sexes, account for a greater proportion of the Bone ALP variation in

  8. 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.

  9. Prognostic Importance of Serum Alkaline Phosphatase in CKD Stages 3–4 in a Clinical Population

    PubMed Central

    Taliercio, Jonathan J.; Schold, Jesse D.; Simon, James F.; Arrigain, Susana; Tang, Anne; Saab, Georges; Nally, Joseph V.; Navaneethan, Sankar D.

    2013-01-01

    Background Elevated total serum alkaline phosphatase (ALP) levels have been associated with mortality in the general population and in dialysis patients. Study Design Retrospective cohort study. Setting & Participants 28,678 patients with chronic kidney disease (CKD) stages 3 and 4 (estimated glomerular filtration rate [eGFR], 15–59 ml/min/1.73 m2) were identified using the Cleveland Clinic Chronic Kidney Disease Registry. CKD was defined as two eGFR values <60 ml/min/1.73 m2 drawn >90 days apart using the Chronic Kidney Disease Epidemiology Collaboration creatinine equation. Predictor ALP levels measured using the calorimetric assay was examined as quartiles (quartile 1, <66 U/L; Q2, 66–81 U/L; Q3, 82–101 U/L; and Q4, ≥102 U/L) and as a continuous measure. Outcomes & Measurements All-cause mortality and ESRD were ascertained using the Social Security Death Index and US Renal Data System. Results After a median follow up of 2.2 years, 588 patients progressed to ESRD and 4,755 died. There was a graded increase in the risk for mortality with higher ALP quartiles (Q2, Q3, Q4) when compared to the reference quartile (Q1) after adjusting for demographics, comorbid conditions, use of relevant medications and liver function tests. The highest quartile of ALP was associated with a hazard ratio for ESRD of 1.38 (95% CI, 1.09–1.76). Each 1-standard deviation (42.7 U/L) higher ALP level was associated with 15% (95% CI, 1.09–1.22) and 16% (95% CI, 1.14–1.18) increased risk of ESRD and mortality respectively. Limitations Single center observational study, lack complete data including PTH for all study participants and attrition bias. Conclusions Higher serum ALP levels in CKD stages 3–4 were independently associated with all-cause mortality and ESRD. PMID:23769134

  10. 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.

  11. Ozone measurements along vertical transects in the Alps.

    PubMed

    Werner, H; Kirchner, M; Welzl, G; Hangartner, M

    1999-01-01

    To investigate the vertical profiles of air pollutants in the boundary layer, aircraft and balloon-born measurements and measurements using a cable car as an instrument platform have been performed in different parts of the Alps. This on-line monitoring of atmospheric pollutants requires expensive and sophisticated techniques. In order to control ambient air quality in remote regions, where no infrastructure like power supply is available, simple instruments are required. The objective of this study, which was coordinated and evaluated by the GSF-Forschungszentrum für Umwelt und Gesundheit was first, to investigate the vertical distribution of ozone in different parts of the Alps and secondly, in addition to continuous analyser measurements, to test monitoring by means of two types of passive samplers. The selection of these samplers - one for one week use and another one for two week application - was based on a passive sampler intercomparison done in a preliminary study one year earlier.

  12. 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.

  13. 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.

  14. Metal-phosphate binders

    SciTech Connect

    Howe, Beth Ann; Chaps-Cabrera, Jesus Guadalupe

    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.

  15. 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.

  16. 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

  17. 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

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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

  3. ALPtraum: ALP production in proton beam dump experiments

    NASA Astrophysics Data System (ADS)

    Döbrich, Babette; Jaeckel, Joerg; Kahlhoefer, Felix; Ringwald, Andreas; Schmidt-Hoberg, Kai

    2016-02-01

    With their high beam energy and intensity, existing and near-future proton beam dumps provide an excellent opportunity to search for new very weakly coupled particles in the MeV to GeV mass range. One particularly interesting example is a so-called axion-like particle (ALP), i.e. a pseudoscalar coupled to two photons. The challenge in proton beam dumps is to reliably calculate the production of the new particles from the interactions of two composite objects, the proton and the target atoms. In this work we argue that Primakoff production of ALPs proceeds in a momentum range where production rates and angular distributions can be determined to sufficient precision using simple electromagnetic form factors. Reanalysing past proton beam dump experiments for this production channel, we derive novel constraints on the parameter space for ALPs. We show that the NA62 experiment at CERN could probe unexplored parameter space by running in `dump mode' for a few days and discuss opportunities for future experiments such as SHiP.

  4. Variation trend of snowmelt runoff in the Japanese Alps catchment

    NASA Astrophysics Data System (ADS)

    Suzuki, K.

    2015-12-01

    The Japanese Alps region is known to experience some of the heaviest snowfall in the world. In this region, precipitation brought by snowfall is more important as a water resource than rainfall. This region experiences exceptionally heavy snowfall that is extreme even by world standard, and in spring, the melting snow becomes a valuable water resource for the region. Snow plays the role of a natural white dam by accumulating in watersheds during winter. Recent studies have reported that the amount of snowfall in Japan will decrease as a result of global warming. However, these studies used data observed at low altitudes. The question arises whether the same theory can be applied to high-altitude mountain areas. Observations of the amount of snow have not been carried out in high-altitude mountain in Japan where the temperature is colder than the threshold temperature of snow/rain even with the temperature rise observed in recent years. Therefore, we cannot discuss the effect of global warming on the change in the amount of snow in the mountainous region of Japan based on observation data. Therefore, in this study, we discuss the relationship between snowmelt runoff and the amount of snow using observation data for the Japanese Alps catchment and present the results of some meteorological observations we carried out at high-altitude sites in the Japanese Alps region.

  5. 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. PMID:25953534

  6. 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.

  7. 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.

  8. Phosphate solubilization potential and phosphatase activity of rhizospheric trichoderma spp.

    PubMed

    Anil, Kapri; Lakshmi, Tewari

    2010-07-01

    Trichoderma sp., a well known biological control agent against several phytopathogens, was tested for its phosphate (P) solubilizing potential. Fourteen strains of Trichoderma sp. were isolated from the forest tree rhizospheres of pinus, deodar, bamboo, guava and oak on Trichoderma selective medium. The isolates were tested for their in-vitro P-solubilizing potential using National Botanical Research Institute Phosphate (NBRIP) broth containing tricalcium phosphate (TCP) as the sole P source, and compared with a standard culture of T. harzianum. All the cultures were found to solubilize TCP but with varying potential. The isolate DRT-1 showed maximum amount of soluble phosphate (404.07 εg.ml(-1)), followed by the standard culture of T. harzianum (386.42 εg.ml(-1)) after 96 h of incubation at 30±1(0)C. Extra-cellular acid and alkaline phosphatases of the fungus were induced only in the presence of insoluble phosphorus source (TCP). High extra-cellular alkaline phosphatase activity was recorded for the isolate DRT-1 (14.50 U.ml(-1)) followed by the standard culture (13.41 U.ml(-1)) at 72h. The cultures showed much lesser acid phosphatase activities. Under glasshouse conditions, Trichoderma sp. inoculation increased chickpea (Cicer arietinum) growth parameters including shoot length, root length, fresh and dry weight of shoot as well as roots, in P-deficient soil containing only bound phosphate (TCP). Shoot weight was increased by 23% and 33% by inoculation with the isolate DRT-1 in the soil amended with 100 and 200 mg TCP kg(-1) soil, respectively, after 60 d of sowing. The study explores high P-solubilizing potential of Trichoderma sp., which can be exploited for the solubilization of fixed phosphates present in the soil, thereby enhancing soil fertility and plant growth.

  9. Effects of the surface characteristics of nano-crystalline and micro-particle calcium phosphate/chitosan composite films on the behavior of human mesenchymal stem cells in vitro.

    PubMed

    Lee, Y-T; Yu, B-Y; Shao, H-J; Chang, C-H; Sun, Y-M; Liu, H-C; Hou, S-M; Young, T-H

    2011-01-01

    Calcium phosphate (CaP) compounds, the main inorganic constituent of mammalian bone tissues, are believed to support bone precursor cell growth and osteogenic differentiation. Chitosan, a deacetylated derivative of chitin, is a versatile biopolymer to offer broad possibilities for cell-based tissue engineering. In the present study, different scales of CaP crystals on chitosan membranes were prepared for culture of human mesenchymal stem cells (hMSCs) in vitro. A series of aqueous CaP suspensions with different concentrations were mixed with chitosan solution and chitosan/calcium phosphate (C/CaP) films were fabricated by the solvent-casting method. With different weight ratios of CaP in chitosan solution, the various surface characteristics of nano-amorphous (C/CaP 0.1), nano-crystalline (C/CaP 0.5) and micro-particle (C/CaP 2) CaP compounds were examined by scanning electron microscopy and electron dispersion spectroscopy. X-ray diffraction on micro-particles of CaP indicated the formation of crystalline hydroxyapatite. The behavior of hMSCs, including proliferation, cell spreading and osteogenic differentiation, was studied on the C/CaP films. In basal culture medium, the incorporation of CaP into chitosan films could promote the proliferation of hMSCs. The C/CaP 0.5 film with connected CaP nano-crystals had better cellular viability. The fluorescence microscope images at 14 days of culture revealed extensive networks of F-actin filaments of hMSCs on chitosan, C/CaP 0.1 and C/CaP 0.5 films. The cellular morphology on C/CaP 2 film with discrete CaP micro-particles was partly restrained. In osteogenic medium, the alkaline phosphatase (ALP) activity of hMSCs increased and showed the process of osteogenic differentiation. The ALP levels on C/CaP 2 film were higher than those on C/CaP 0.1 and C/CaP 0.5 films. These results demonstrated that the crystallinity and topography of CaP on chitosan membranes could modulate the behaviors of cultured hMSCs in vitro.

  10. 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.

  11. 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.

  12. 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.

  13. 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

  14. 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

  15. New, rare or remarkable microfungi in the Italian Alps (Carnic Alps)--part II--other microfungi.

    PubMed

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

    2004-01-01

    In addition to the collection of Ascomycotina in the Carnic Alps (see New, rare or remarkable microfungi in the Italian Alps (Carnic Alps) part I ) we were able to treasure about 300 species of parasitic and saprophytic microfungi. Among them Basidiomycetes, Ascomycetes and Deuteromycets like Bostrichonema polygoni (UNGER) SCHROT. on Polygonum viviparum L., Chrysomyxa rhododendri DE BY on Picea abies (L.) KARSTEN, Coleosporium tussilaginis (PERS.) BERK. I=C. cacaliae OTTH.] on Adenostyles glabra (MILL.) DC., Dasyscyphus barbatus (KUNZE) MASSEE on Lonicera nigra L., Leptosphaeria coniothyrium (FUCKEL) SACC. on Rosa canina L., Leptotrochila brunellae (LIND) DENNIS on Prunella grandiflora (L.) SCHOLLER., Marssonina kriegeriana (BES.) MAGNUS on Salix reticulata L., Puccinia alpina FUCKEL on Viola biflora L., Puccinia maculosa (STRAUSS.) ROHLING and Erysiphe cichoracearum DC. On Prenanthes purpurea L., Septoria microsora SPENG. on Gentianella germanica (WILLD.) BORNER, Urocystis orobranches (FR.) FISCH. V. WALDH. on Orobranche gracilis SM., Urocystis violae (J. SOWERBY) A. FISCHER VON WALDHEIN on Viola biflora L. and Uromyces phyteumatum (DC.) UNG. on Phyteuma spicatum L. were dominant. All samples are located in the Herbarium ESS Mycotheca Parva, Collection G.B. Feige/N. Ale-Agha.

  16. 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.

  17. Association of Cry1Ac Toxin Resistance in Helicoverpa zea (Boddie) with Increased Alkaline Phosphatase Levels in the Midgut Lumen

    PubMed Central

    Caccia, Silvia; Moar, William J.; Chandrashekhar, Jayadevi; Oppert, Cris; Anilkumar, Konasale J.; Jurat-Fuentes, Juan Luis

    2012-01-01

    Resistance to Bacillus thuringiensis Cry1Ac toxin was characterized in a population of Helicoverpa zea larvae previously shown not to have an alteration in toxin binding as the primary resistance mechanism to this toxin. Cry1Ac-selected larvae (AR1) were resistant to protoxins and toxins of Cry1Ab, Cry1Ac, and the corresponding modified proteins lacking helix α-1 (Cry1AbMod and Cry1AcMod). When comparing brush border membrane vesicles (BBMVs) prepared from susceptible (LC) and AR1 larval midguts, there were only negligible differences in overall Cry1Ac toxin binding, though AR1 had 18% reversible binding, in contrast to LC, in which all binding was irreversible. However, no differences were detected in Cry1Ac-induced pore formation activity in BBMVs from both strains. Enzymatic activities of two putative Cry1Ac receptors (aminopeptidase N [APN] and alkaline phosphatase [ALP]) were significantly reduced (2-fold and 3-fold, respectively) in BBMVs from AR1 compared to LC larvae. These reductions corresponded to reduced protein levels in midgut luminal contents only in the case of ALP, with an almost 10-fold increase in specific ALP activity in midgut fluids from AR1 compared to LC larvae. Partially purified H. zea ALP bound Cry1Ac toxin in ligand blots and competed with Cry1Ac toxin for BBMV binding. Based on these results, we suggest the existence of at least one mechanism of resistance to Cry1A toxins in H. zea involving binding of Cry1Ac toxin to an ALP receptor in the larval midgut lumen of resistant larvae. PMID:22685140

  18. Association of Cry1Ac toxin resistance in Helicoverpa zea (Boddie) with increased alkaline phosphatase levels in the midgut lumen.

    PubMed

    Caccia, Silvia; Moar, William J; Chandrashekhar, Jayadevi; Oppert, Cris; Anilkumar, Konasale J; Jurat-Fuentes, Juan Luis; Ferré, Juan

    2012-08-01

    Resistance to Bacillus thuringiensis Cry1Ac toxin was characterized in a population of Helicoverpa zea larvae previously shown not to have an alteration in toxin binding as the primary resistance mechanism to this toxin. Cry1Ac-selected larvae (AR1) were resistant to protoxins and toxins of Cry1Ab, Cry1Ac, and the corresponding modified proteins lacking helix α-1 (Cry1AbMod and Cry1AcMod). When comparing brush border membrane vesicles (BBMVs) prepared from susceptible (LC) and AR1 larval midguts, there were only negligible differences in overall Cry1Ac toxin binding, though AR1 had 18% reversible binding, in contrast to LC, in which all binding was irreversible. However, no differences were detected in Cry1Ac-induced pore formation activity in BBMVs from both strains. Enzymatic activities of two putative Cry1Ac receptors (aminopeptidase N [APN] and alkaline phosphatase [ALP]) were significantly reduced (2-fold and 3-fold, respectively) in BBMVs from AR1 compared to LC larvae. These reductions corresponded to reduced protein levels in midgut luminal contents only in the case of ALP, with an almost 10-fold increase in specific ALP activity in midgut fluids from AR1 compared to LC larvae. Partially purified H. zea ALP bound Cry1Ac toxin in ligand blots and competed with Cry1Ac toxin for BBMV binding. Based on these results, we suggest the existence of at least one mechanism of resistance to Cry1A toxins in H. zea involving binding of Cry1Ac toxin to an ALP receptor in the larval midgut lumen of resistant larvae. PMID:22685140

  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. 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.

  1. 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

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

    PubMed Central

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

    2010-01-01

    Summary 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. PMID:19930184

  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. Overproduction of acetate kinase activates the phosphate regulon in the absence of the phoR and phoM functions in Escherichia coli.

    PubMed Central

    Lee, T Y; Makino, K; Shinagawa, H; Nakata, A

    1990-01-01

    A DNA fragment of Escherichia coli cloned on pBR322 elevated the production of alkaline phosphatase and phosphate-binding protein in a phoR phoM strain. Nucleotide sequence analysis and enzyme assays revealed that the DNA fragment contained the ackA gene, which codes for acetate kinase. A high gene dosage of ackA was needed to induce the production of alkaline phosphatase and phosphate-binding protein in this strain. Overexpression of ackA elevated the intracellular ATP concentration, an effect that might be related to activation of the phosphate regulon in the phoR phoM strain. Images PMID:2158965

  5. 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.

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

    PubMed

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

    2014-07-16

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

  7. Involvement of AlpV, a New Member of the Streptomyces Antibiotic Regulatory Protein Family, in Regulation of the Duplicated Type II Polyketide Synthase alp Gene Cluster in Streptomyces ambofaciens

    PubMed Central

    Aigle, Bertrand; Pang, Xiuhua; Decaris, Bernard; Leblond, Pierre

    2005-01-01

    A type II polyketide synthase gene cluster located in the terminal inverted repeats of Streptomyces ambofaciens ATCC 23877 was shown to be responsible for the production of an orange pigment and alpomycin, a new antibiotic probably belonging to the angucycline/angucyclinone class. Remarkably, this alp cluster contains five potential regulatory genes, three of which (alpT, alpU, and alpV) encode proteins with high similarity to members of the Streptomyces antibiotic regulatory protein (SARP) family. Deletion of the two copies of alpV (one in each alp cluster located at the two termini) abolished pigment and antibiotic production, suggesting that AlpV acts as a transcriptional activator of the biosynthetic genes. Consistent with this idea, the transcription of alpA, which encodes a ketosynthase essential for orange pigment and antibiotic production, was impaired in the alpV mutant, while the expression of alpT, alpU, and alpZ, another regulatory gene encoding a γ-butyrolactone receptor, was not significantly affected. Real-time PCR experiments showed that transcription of alpV in the wild-type strain increases dramatically after entering the transition phase. This induction precedes that of alpA, suggesting that AlpV needs to reach a threshold level to activate the expression of the structural genes. When introduced into an S. coelicolor mutant with deletions of actII-ORF4 and redD, the SARP-encoding genes regulating the biosynthesis of actinorhodin and undecylprodigiosin, respectively, alpV was able to restore actinorhodin production only. However, actII-ORF4 did not complement the alpV mutant, suggesting that AlpV and ActII-ORF4 may act in a different way. PMID:15774892

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. Isotopic composition of sulfate accumulations, Northern Calcareous Alps, Austria

    NASA Astrophysics Data System (ADS)

    Bojar, Ana-Voica; Halas, Stanislaw; Bojar, Hans-Peter; Trembaczowski, Andrzej

    2015-04-01

    The Eastern Alps are characterised by the presence of three main tectonic units, such as the Lower, Middle and Upper Austroalpine, which overlie the Penninicum (Tollmann, 1977). The Upper Austroalpine unit consists of the Northern Calcareous Alps (NCA) overlying the Greywacke zone and corresponding to the Graz Paleozoic, Murau Paleozoic and the Gurktal Nappe. Evaporitic rocks are lacking in the later ones. The Northern Calcareous Alps are a detached fold and thrust belt. The sedimentation started in the Late Carboniferous or Early Permian, the age of the youngest sediments being Eocene. The NCA are divided into the Bajuvaric, Tirolic and Juvavic nappe complexes. The evaporitic Haselgebirge Formation occurs in connection with the Juvavic nappe complex at the base of the Tirolic units (Leitner et al., 2013). The Haselgebirge Formation consists mainly of salt, shales, gypsum and anhydrite and includes the oldest sediments of the NCA. The age of the Haselgebirge Formation, established by using spors and geochronological data, is Permian to Lower Triassic. For the Northern Calcareous Alps, the mineralogy of sulphate accumulations consists mainly of gypsum and anhydrite and subordonates of carbonates. The carbonates as magnesite, dolomite and calcite can be found either as singular crystals or as small accumulations within the hosting gypsum. Sulfides (sphalerite, galena, pyrite), sulfarsenides (enargite, baumhauerite) and native sulphur enrichments are known from several deposits (Kirchner, 1987; Postl, 1990). The investigated samples were selected from various gypsum and halite rich deposits of the Northern Calcareous Alps. A total of over 20 samples were investigated, and both oxygen and sulfur isotopic composition were determined for anhydrite, gyps, polyhalite, blödite and langbeinite. The sulfur isotopic values vary between 10.1 to 14 ‰ (CDT), with three values higher than 14 ‰. The Oxygen isotopic values show a range from 9 to 23 ‰ (SMOW). The sulfur

  13. Novel high-pressure phases of AlP from first principles

    NASA Astrophysics Data System (ADS)

    Liu, Chao; Hu, Meng; Luo, Kun; Yu, Dongli; Zhao, Zhisheng; He, Julong

    2016-05-01

    By utilizing a crystal structure prediction software via particle swarm optimization, this study proposes three new high-pressure phases of aluminum phosphide (AlP) with high density and high hardness, in addition to previously proposed phases (wz-, zb-, rs-, NiAs-, β-Sn-, CsCl-, and Cmcm-AlP). These new phases are as follows: (1) an I 4 ¯ 3d symmetric structure (cI24-AlP) at 55.2 GPa, (2) an R 3 ¯ m symmetric structure (hR18-AlP) at 9.9 GPa, and (3) a C222 symmetric structure (oC12-AlP) at 20.6 GPa. Based on first-principle calculations, these phases have higher energetic advantage than CsCl- and β-Sn-AlP at ambient pressure. The independent elastic constants and phonon dispersion spectra are calculated to check the mechanical and dynamic stabilities of these phases. According to mechanical property studies, these new AlP phases have higher hardness than NiAs-AlP, and oC12-AlP has the highest hardness of 7.9 GPa. Electronic band structure calculations indicate that NiAs- and hR18-AlP have electrical conductivity. Additionally, wz-, zb-, and oC12-AlP possess semiconductive properties with indirect bandgaps, and cI24-AlP has a semiconductive property with a direct bandgap.

  14. Genetic differentiation among populations of Cicerbita alpina (L.) Wallroth (asteraceae) in the western Alps.

    PubMed

    Michl, Thomas; Huck, Stefan; Haase, Peter; Büdel, Burkhard

    2007-01-01

    In this study we analyzed the genetic population structure of the hygrophilous tall-herb Cicerbita alpina in the western Alps because this group of mountain plants is underrepresented in the biogeographical literature. AFLP (amplified fragment length polymorphism) fingerprints of 40 samples were analyzed from four populations situated in a transect from the southwestern Alps to the eastern part of the western Alps and one population from the Black Forest outside the Alps. Two genetic groups can be distinguished. The first group (A) comprises the populations from the northern and eastern parts of the western Alps, and the second group (B) comprises the populations from the southwestern Alps and the Black Forest. Group A originates most likely from at least one refugium in the southern piedmont regions of the Alps. This result provides molecular evidence for a humid climate at the southern margin of the Alps during the Würm glaciation. Group B originates presumably from western or northern direction and we discuss two possible scenarios for the colonization of the Alps, i.e. (1) long-distance dispersal from southwestern refugia and (2) colonization from nearby refugia in the western and/or northern Alpine forelands. The study demonstrates that the target species harbours considerable genetic diversity, even on a regional scale, and therefore is a suitable model for phylogeographic research.

  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. 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. PMID:26559965

  17. Fundamentals of phosphate transfer.

    PubMed

    Kirby, Anthony J; Nome, Faruk

    2015-07-21

    Historically, the chemistry of phosphate transfer-a class of reactions fundamental to the chemistry of Life-has been discussed almost exclusively in terms of the nucleophile and the leaving group. Reactivity always depends significantly on both factors; but recent results for reactions of phosphate triesters have shown that it can also depend strongly on the nature of the nonleaving or "spectator" groups. The extreme stabilities of fully ionised mono- and dialkyl phosphate esters can be seen as extensions of the same effect, with one or two triester OR groups replaced by O(-). Our chosen lead reaction is hydrolysis-phosphate transfer to water: because water is the medium in which biological chemistry takes place; because the half-life of a system in water is an accepted basic index of stability; and because the typical mechanisms of hydrolysis, with solvent H2O providing specific molecules to act as nucleophiles and as general acids or bases, are models for reactions involving better nucleophiles and stronger general species catalysts. Not least those available in enzyme active sites. Alkyl monoester dianions compete with alkyl diester monoanions for the slowest estimated rates of spontaneous hydrolysis. High stability at physiological pH is a vital factor in the biological roles of organic phosphates, but a significant limitation for experimental investigations. Almost all kinetic measurements of phosphate transfer reactions involving mono- and diesters have been followed by UV-visible spectroscopy using activated systems, conveniently compounds with good leaving groups. (A "good leaving group" OR* is electron-withdrawing, and can be displaced to generate an anion R*O(-) in water near pH 7.) Reactivities at normal temperatures of P-O-alkyl derivatives-better models for typical biological substrates-have typically had to be estimated: by extended extrapolation from linear free energy relationships, or from rate measurements at high temperatures. Calculation is free

  18. 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

  19. Localization of d-myoinositol 1:2-cyclic phosphate 2-phosphohydrolase in rat kidney

    PubMed Central

    Clarke, N.; Dawson, R. M. C.

    1972-01-01

    1. On subcellular fractionation of rat kidney homogenates by differential and density-gradient centrifugation, the bulk of the inositol 1:2-cyclic phosphate 2-phosphohydrolase activity remains with the alkaline phosphatase activity, suggesting localization in the brush borders of the proximal tubules. 2. Histochemical studies with a medium containing inositol 1:2-cyclic phosphate and Escherichia coli phosphomonoesterase show Gomori staining around the brush borders of the proximal tubules in the outer cortex only. 3. Serial sections across the kidney from cortex perimeter to papilla suggest that the inositol 1:2-cyclic phosphate 2-phosphohydrolase has a limited distribution along the proximal tubule of the nephron, probably being limited to the pars convoluta, whereas the alkaline phosphatase extends along the pars recta. ImagesPLATE 1PLATE 2 PMID:4347783

  20. Precipitation and snow cover variability in the french alps

    NASA Astrophysics Data System (ADS)

    Martin, Eric; Durand, Yves

    The distribution of winter precipitation as analysed by the meteorological analysis system SAFRAN is validated using data from two test sites. This system, applied to the French Alps, shows that the frequency of high precipitation events is not necessarily linked to mean precipitation. Using a downscaling procedure, the system was run with General Circulation Model (GCM) outputs. The corresponding snow cover is derived with the snow model CROCUS. The results are very sensitive to the quality of the GCM run. The analyses of two future climate scenarios show that drastic changes in precipitation distribution may occur in the future.

  1. 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

  2. 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

  3. 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. PMID:27035384

  4. 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.

  5. Ribose-5-phosphate isomerase and ribulose-5-phosphate kinase show apparent specificity for a specific ribulose 5-phosphate species.

    PubMed

    Anderson, L E

    1987-02-01

    Ribose-5-phosphate isomerase and ribulose-5-phosphate kinase appear to show specificity for a particular ribulose 5-phosphate species. The effect of this specificity will be channeling of ribulose 5-phosphate from the isomerase to the kinase during photosynthesis.

  6. 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.

  7. 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.

  8. 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 than two times the surface areas of reference cathodes and therefore offered the best potential for improved performance.

  9. 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.

  10. 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)

  11. Average snowcover density values in Eastern Alps mountain

    NASA Astrophysics Data System (ADS)

    Valt, M.; Moro, D.

    2009-04-01

    The Italian Avalanche Warning Services monitor the snow cover characteristics through networks evenly distributed all over the alpine chain. Measurements of snow stratigraphy and density are very frequently performed with sampling rates of 1 -2 times per week. Snow cover density values are used to compute the dimensions of the building roofs as well as to design avalanche barriers. Based on the measured snow densities the Electricity Board can predict the amount of water resources deriving from snow melt in high relieves drainage basins. In this work it was possible to compute characteristic density values of the snow cover in the Eastern Alps using the information contained in the database from the ARPA (Agenzia Regionale Protezione Ambiente)-Centro Valanghe di Arabba, and Ufficio Valanghe- Udine. Among the other things, this database includes 15 years of stratigraphic measurements. More than 6,000 snow stratigraphic logs were analysed, in order to derive typical values as for geographical area, altitude, exposure, snow cover thickness and season. Computed values were compared to those established by the current Italian laws. Eventually, experts identified and evaluated the correlations between the seasonal variations of the average snow density and the variations related to the snowfall rate in the period 1994-2008 in the Eastern Alps mountain range

  12. Using Structural Thermochronology to Reconstruct Backfolding in the Swiss Alps

    NASA Astrophysics Data System (ADS)

    McPhillips, D.; Brandon, M. T.; Vance, J. A.

    2006-12-01

    A new interpretational method takes advantage of geometric relationships among zircon fission track ages and allows direct dating of the formation and propagation of the retroshear zone of the Swiss Alps. The retroshear zone is defined by a 20 km wide, south-vergent "backfold", formed as part of the doubly-vergent structure of the orogen. Our "structural thermochronologic" analysis relies on the concept of the isochrone, which is a surface in the earth that everywhere has the same cooling age. Isochrone surfaces are formed as rocks pass through the closure isotherm. They tend to have very gentle to flat initial dips, and they can be used, like bedded stratigraphy, to track deformation. The age gradient across uniformly dipping isochrones provides a measure of exhumation rate in datasets where typical age-elevation relationships are scattered. In the Alps, isochrone-derived exhumation rates increase more than fourfold at 16 Ma, from 0.25 km/Ma to ~ 1.0 km/Ma. Isochrones dip uniformly above and below the increase, at about 10 degrees toward the southeast. The change in exhumation rate corresponds generally to the Insubric line. Where exhumation rates are high, to the north of the Insubric line, cooling ages are Miocene, while exhumation rates to the south are low and cooling ages are Cretaceous to Triassic. We use a kinematic folding model to show that the change in exhumation rates and the uniform southward dips are both a result of steady southward propagation of the Alpine backfold.

  13. Alkaline Water and Longevity: A Murine Study.

    PubMed

    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.

  14. 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.

  15. Grace DAKASEP alkaline battery separator

    NASA Technical Reports Server (NTRS)

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

    1987-01-01

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

  16. The design of alkaline fuel cells

    NASA Astrophysics Data System (ADS)

    Strasser, K.

    1990-01-01

    Alkaline fuel cells recently developed have yielded satisfactory operation even in the cases of their use of mobile and matrix-type electrolytes; the advantages of realistic operation have been demonstrated by a major West German manufacturer's 100 kW alkaline fuel cell apparatus, which was operated in the role of an air-independent propulsion system. Development has begun for a spacecraft alkaline fuel cell of the matrix-electrolyte configuration.

  17. Controlled intracellular proteolysis during postpartal involution of the uterus: characterization and regulation of an alkaline proteinase.

    PubMed

    Roth, M; Hoechst, M; Afting, E G

    1981-01-01

    The postpartal involution of the uterus is predominantly due to cellular hypotrophy. This implies an intracellular proteolytic system which must be carefully controlled pre and post partum. We have characterized and partially purified a proteinase with an alkaline pH-optimum of activity and a proteinase inhibitor protein which inhibits this proteinase very strongly. The alkaline proteinase copurifies with the actomyosin complex of the uterine myometrium and degrades the actomyosin complex with a concomitant loss of its myosin-ATPase activity. The alkaline proteinase is a very labile enzyme, markedly sensitive to SH-group modifying agents and has very high molecular weight at the present state of purification. This proteolytic enzyme could specifically be separated from the main components of the actomyosin complex by extraction with low ionic strength phosphate buffers. The proteinase inhibitor protein may control the activity of this alkaline proteinase during pregnancy and involution. The inhibitor protein raises 15-fold during pregnancy, possibly blocks important steps of intracellular proteolysis and permits organ growth. The dramatic fall of the inhibitor protein activity after parturition, which precedes the loss of weight, could release the proteolytic system, including the alkaline proteinase, and permits controlled intracellular degradation.

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

    PubMed

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

    2015-07-15

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

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

    PubMed

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

    2015-06-01

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

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

    PubMed

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

    2015-06-01

    MicroRNAs (MiRNAs) have been regarded as clinically important biomarkers and drug discovery targets. In this work, we reported a simple and ultrasensitive electrochemical method for miRNAs detection based on single enzyme amplification and electrochemical-chemical-chemical (ECC) redox cycling. Specifically, upon contact with the target miRNAs, the hairpin structure of biotinylated DNA immobilized on gold electrode was destroyed and the biotin group in DNA was forced away from the electrode surface, allowing for the coupling of streptavidin-conjugated alkaline phosphatase (SA-ALP). Then, ascorbic acid (AA, the enzymatic product of ALP) triggered the ECC redox cycling with ferrocene methanol (FcM) and tris(2-carboxyethyl)phosphine (TCEP) as the redox mediator and the chemical reducing reagent, respectively. The method was more sensitive than that with horseradish peroxidase (HRP) or glucose oxidase (GOx) triggered recycling since one ALP molecule captured by one target miRNA molecule promoted the production of thousands of AA. Analytical merits (e.g., detection limit, dynamic range, specificity, regeneration and reproducibility) were evaluated. The feasibility of the method for analysis of miRNA-21 in human serum has also been demonstrated. PMID:26002330

  1. 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.

  2. 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.

  3. 21 CFR 184.1434 - Magnesium phosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Magnesium phosphate. 184.1434 Section 184.1434... Listing of Specific Substances Affirmed as GRAS § 184.1434 Magnesium phosphate. (a) Magnesium phosphate includes both magnesium phosphate, dibasic, and magnesium phosphate, tribasic. Magnesium phosphate,...

  4. 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,...

  5. 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,...

  6. 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....

  7. 21 CFR 184.1434 - Magnesium phosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Magnesium phosphate. 184.1434 Section 184.1434 Food... Specific Substances Affirmed as GRAS § 184.1434 Magnesium phosphate. (a) Magnesium phosphate includes both magnesium phosphate, dibasic, and magnesium phosphate, tribasic. Magnesium phosphate, dibasic...

  8. Effects of dietary vitamin E on mucosal maltase and alkaline phosphatase enzyme activities and on the amount of mucosal malonyldialdehyde in broiler chickens

    PubMed Central

    Farrokhifar, Seyed Hamid; Ali Jafari, Ramezan; Erfani Majd, Naeem; Fatemi Tabatabaee, Seyed Reza; Mayahi, Mansour

    2013-01-01

    The effects of dietary vitamin E levels on mucosal maltase and alkaline phosphatase (ALP) enzyme activities and on the amount of mucosal malonyldialdehyde (MDA) in broiler chickens were studied in the present study. One hundred and eighty of male day old broiler chicks (Ross 308 strain) were randomly assigned into five groups, each with three replicates and 12 chicks in each replicate. Chickens in group A were fed corn-soy- based diet, while those in groups B, C, D and E were fed the same diet with 20, 60, 180, and 540 mg kg-1 vitamin E supplement (d-alpha tocopherol), respectively. Six birds were randomly chosen from each group, and were euthanized on days 10, 21, 32, and 42 of age. One segment of small intestine outset was homogenized and mucosal ALP and maltase activity were measured. Moreover, mucosal lipid peroxidate amount was measured to reveal the impact of vitamin E on oxidative stress. Maltase activity was increased with the increase of vitamin E up to 60 mg kg-1 of diet while with further levels, it was decreased. Addition of 60 mg kg-1 of vitamin E to the diet significantly increased ALP enzyme activity (p ≤ 0.001). Addition of 540 mg kg-1 of vitamin E supplement to the diet led to the minimum amount of MDA at 32 days of age. It may be concluded that supplementation of broiler's diet with 60 mg kg-1 of vitamin E can increase mucosal maltase and ALP enzyme activity. PMID:25568675

  9. Effects of hydrogen sulfide on the expression of alkaline phosphatase, osteocalcin and collagen type I in human periodontal ligament cells induced by tension force stimulation.

    PubMed

    Qin, Jing; Hua, Yongmei

    2016-10-01

    Periodontal ligament cells (PDLCs) are important in homeostasis and remodeling in the mechanically‑stimulated periodontium. The aim of the present study was to investigate the effects of hydrogen sulfide (H2S) on periodontal tissue remodeling by examining the mRNA and protein expression levels of alkaline phosphatase (ALP), osteocalcin (OCN) and collagen type I (COL‑1) in human (h)PDLCs induced by tension force application. Cultured hPDLCs were treated with H2S for 24 h, followed by application of a tension force for 1, 3 and 6 h. Cell proliferation and apoptosis were determined using a Cell Counting Kit 8 assay and flow cytometric analysis, respectively. The mRNA expression levels of ALP, OCN and COL‑1 were quantified using reverse transcription‑quantitative polymerase chain reaction analysis, and western blot analysis was used to detect the protein levels of ALP, OCN and COL‑1. The results demonstrated that the mRNA and protein expression levels of ALP, OCN and COL‑1 increased with H2S treatment in a concentration‑dependent manner, which was enhanced by the application of tension force in a relatively short period of time. These findings suggested that H2S may be important in periodontal tissue remodeling during orthodontic tooth movement via increasing hPDLC differentiation, tissue mineralization, bone formation and collagen synthesis. PMID:27573279

  10. Phylogenetic Analysis Identifies Many Uncharacterized Actin-like Proteins (Alps) in Bacteria: Regulated Polymerization, Dynamic Instability, and Treadmilling in Alp7A

    PubMed Central

    Derman, Alan I.; Becker, Eric C.; Truong, Bao D.; Fujioka, Akina; Tucey, Timothy M.; Erb, Marcella L.; Patterson, Paula C.; Pogliano, Joe

    2010-01-01

    Summary Actin, one of the most abundant proteins in the eukaryotic cell, also has an abundance of relatives in the eukaryotic proteome. To date though, only five families of actins have been characterized in bacteria. We have conducted a phylogenetic search and uncovered more than 35 highly divergent families of actin-like proteins (Alps) in bacteria. Their genes are found primarily on phage genomes, on plasmids, and on integrating conjugative elements, and are likely to be involved in a variety of functions. We characterize three Alps and find that all form filaments in the cell. The filaments of Alp7A, a plasmid partitioning protein and one of the most divergent of the Alps, display dynamic instability and also treadmill. Alp7A requires other elements from the plasmid to assemble into dynamic polymers in the cell. Our findings suggest that most if not all of the Alps are indeed actin relatives, and that actin is very well represented in bacteria. PMID:19602153

  11. Phosphatidylcholine affects the secretion of the alkaline phosphatase PhoA in Pseudomonas strains.

    PubMed

    Liu, Xin; Long, Deliang; You, Heng; Yang, Dingpeng; Zhou, Shuang; Zhang, Shuting; Li, Mengqiu; He, Miao; Xiong, Min; Wang, Xingguo

    2016-11-01

    Pseudomonas aeruginosa ATCC 27853 and Pseudomonas sp. 593 use the phosphatidylcholine synthase pathway (Pcs-pathway) for the biosynthesis of phosphatidylcholine (PC). Both bacterial strains contain the phoA and lapA genes encoding alkaline phosphatases (ALP) and display strong ALP activities. The PhoA and LapA enzymes are thought to be independently secreted via the Xcp and Hxc type II secretion system (T2SS) subtypes, in which the Hxc system may act as a complementary mechanism when the Xcp pathway becomes limiting. Inactivation of the pcs gene in both bacteria abolished PC synthesis and resulted in approximately 50% less ALP activity in the cell-free culture. Analysis by western blotting showed that LapA protein content in the wild type and the pcs- mutant was unchanged in the cytoplasmic, periplasmic or extracellular protein fractions. In contrast, the PhoA protein in the pcs- mutant was less prevalent among extracellular proteins but was more abundant in the periplasmic protein fraction compared to the wild type. Semi- quantitative reverse transcriptase PCR showed that phoA, lapA and 12 xcp genes were equally expressed at the transcriptional level in both the wild types and the pcs- mutants. Our results demonstrate that the absence of PC in bacterial membrane phospholipids does not interfere with the transcription of the phoA and lapA genes but primarily affects the export of PhoA from the cytoplasm to the extracellular environment via the Xcp T2SS.

  12. Phosphatidylcholine affects the secretion of the alkaline phosphatase PhoA in Pseudomonas strains.

    PubMed

    Liu, Xin; Long, Deliang; You, Heng; Yang, Dingpeng; Zhou, Shuang; Zhang, Shuting; Li, Mengqiu; He, Miao; Xiong, Min; Wang, Xingguo

    2016-11-01

    Pseudomonas aeruginosa ATCC 27853 and Pseudomonas sp. 593 use the phosphatidylcholine synthase pathway (Pcs-pathway) for the biosynthesis of phosphatidylcholine (PC). Both bacterial strains contain the phoA and lapA genes encoding alkaline phosphatases (ALP) and display strong ALP activities. The PhoA and LapA enzymes are thought to be independently secreted via the Xcp and Hxc type II secretion system (T2SS) subtypes, in which the Hxc system may act as a complementary mechanism when the Xcp pathway becomes limiting. Inactivation of the pcs gene in both bacteria abolished PC synthesis and resulted in approximately 50% less ALP activity in the cell-free culture. Analysis by western blotting showed that LapA protein content in the wild type and the pcs- mutant was unchanged in the cytoplasmic, periplasmic or extracellular protein fractions. In contrast, the PhoA protein in the pcs- mutant was less prevalent among extracellular proteins but was more abundant in the periplasmic protein fraction compared to the wild type. Semi- quantitative reverse transcriptase PCR showed that phoA, lapA and 12 xcp genes were equally expressed at the transcriptional level in both the wild types and the pcs- mutants. Our results demonstrate that the absence of PC in bacterial membrane phospholipids does not interfere with the transcription of the phoA and lapA genes but primarily affects the export of PhoA from the cytoplasm to the extracellular environment via the Xcp T2SS. PMID:27664720

  13. 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.

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. Dysregulation of phosphate metabolism and conditions associated with phosphate toxicity

    PubMed Central

    Brown, Ronald B; Razzaque, Mohammed S

    2015-01-01

    Phosphate homeostasis is coordinated and regulated by complex cross-organ talk through delicate hormonal networks. Parathyroid hormone (PTH), secreted in response to low serum calcium, has an important role in maintaining phosphate homeostasis by influencing renal synthesis of 1,25-dihydroxyvitamin D, thereby increasing intestinal phosphate absorption. Moreover, PTH can increase phosphate efflux from bone and contribute to renal phosphate homeostasis through phosphaturic effects. In addition, PTH can induce skeletal synthesis of another potent phosphaturic hormone, fibroblast growth factor 23 (FGF23), which is able to inhibit renal tubular phosphate reabsorption, thereby increasing urinary phosphate excretion. FGF23 can also fine-tune vitamin D homeostasis by suppressing renal expression of 1-alpha hydroxylase (1α(OH)ase). This review briefly discusses how FGF23, by forming a bone–kidney axis, regulates phosphate homeostasis, and how its dysregulation can lead to phosphate toxicity that induces widespread tissue injury. We also provide evidence to explain how phosphate toxicity related to dietary phosphorus overload may facilitate incidence of noncommunicable diseases including kidney disease, cardiovascular disease, cancers and skeletal disorders. PMID:26131357

  19. Dysregulation of phosphate metabolism and conditions associated with phosphate toxicity.

    PubMed

    Brown, Ronald B; Razzaque, Mohammed S

    2015-01-01

    Phosphate homeostasis is coordinated and regulated by complex cross-organ talk through delicate hormonal networks. Parathyroid hormone (PTH), secreted in response to low serum calcium, has an important role in maintaining phosphate homeostasis by influencing renal synthesis of 1,25-dihydroxyvitamin D, thereby increasing intestinal phosphate absorption. Moreover, PTH can increase phosphate efflux from bone and contribute to renal phosphate homeostasis through phosphaturic effects. In addition, PTH can induce skeletal synthesis of another potent phosphaturic hormone, fibroblast growth factor 23 (FGF23), which is able to inhibit renal tubular phosphate reabsorption, thereby increasing urinary phosphate excretion. FGF23 can also fine-tune vitamin D homeostasis by suppressing renal expression of 1-alpha hydroxylase (1α(OH)ase). This review briefly discusses how FGF23, by forming a bone-kidney axis, regulates phosphate homeostasis, and how its dysregulation can lead to phosphate toxicity that induces widespread tissue injury. We also provide evidence to explain how phosphate toxicity related to dietary phosphorus overload may facilitate incidence of noncommunicable diseases including kidney disease, cardiovascular disease, cancers and skeletal disorders. PMID:26131357

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

    PubMed

    Thengodkar, Rutwik Ravindra Mandakini; Sivakami, S

    2010-07-01

    Spirulina is a photosynthetic, filamentous, spiral-shaped, multicellular, blue-green microalga. The two most important species are Spirulina maxima and Spirulina platensis. Spirulina is considered an excellent food, lacking toxicity and having corrective properties against viral attacks, anemia, tumor growth and malnutrition. We have observed that cultures of Spirulina platensis grow in media containing up to 80 ppm of the organophosphorous pesticide, Chlorpyrifos. It was found to be due to an alkaline phosphatase (ALP) activity that was detected in cell free extracts of Spirulina platensis. This activity was purified from the cell free extracts using ammonium sulphate precipitation and gel filtration and shown to belong to the class of EC 3.1.3.1 ALP. The purified enzyme degrades 100 ppm Chlorpyrifos to 20 ppm in 1 h transforming it into its primary metabolite 3, 5, 6-trichloro-2-pyridinol. This is the first report of degradation of Chlorpyrifos by Spirulina platensis whose enzymic mechanism has been clearly identified. These findings have immense potential for harnessing Spirulina platensis in bioremediation of polluted ecosystems. PMID:20127145

  1. 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.

  2. Morphological expression of active tectonics in the Southern Alps

    NASA Astrophysics Data System (ADS)

    Robl, Jörg; Heberer, Bianca; Neubauer, Franz; Hergarten, Stefan

    2015-04-01

    Evolving drainage pattern and corresponding metrics of the channels (e.g. normalized steepness index) are sensitive indicators for tectonic or climatic events punctuating the evolution of mountain belts and their associated foreland basins. The analysis of drainage systems and their characteristic properties represents a well-established approach to constrain the impact of tectonic and climatic drivers on mountainous landscapes in the recent past. The Southern Alps (SA) are one of the seismically most active zones in the periphery of northern Adria. Recent deformation is caused by the ongoing convergence of the Adriatic and European plate and is recorded by numerous earthquakes in the domain of the SA. Deformation in the SA is characterized by back-thrusting causing crustal thickening and should therefore result in uplift and topography formation. The vertical velocity field determined by GPS-data clearly indicates a belt of significant uplift in the south South alpine indenter between Lake Garda in the west and the Triglav in the east and strong subsidence of the foreland basin surrounding the Mediterranean Sea near Venice, although subsidence is often related to ongoing subduction of the Adriatic microplate underneath Appennines. Despite of these short term time series, timing, rates and drivers of alpine landscape evolution are not well constrained and the linkage between crustal deformation and topographic evolution of this highly active alpine segment remains unclear for the following reasons: (1) The eastern Southern Alps were heavily overprinted by the Pleistocene glaciations and tectonic signals in the alpine landscape are blurred. Only the transition zone to the southern foreland basin remained unaffected and allows an analysis of a glacially undisturbed topography. (2) The major part of this domain is covered by lithology (carbonatic rocks) which is unsuitable for low temperature geochronology and cosmogenic isotope dating so that exhumation and erosion

  3. 25Al+p Elastic Scattering with CRIB

    NASA Astrophysics Data System (ADS)

    Pearson, Jonty; Chen, A. A.; Kubono, S.; Yamaguchi, H.; He, J. J.; Khlem, L. H.; Salto, A.; Amadio, G.; Fajikawa, H.; Nilkura, M.; Wakabayashi, Y.; Teranishi, T.; Nishimura, S.; Togano, Y.; Odahara, A.; Moon, J. Y.; Kwon, Y. K.; Cherubini, S.; Pizzone, R.; La Cognata, M.

    The present rate of the 25 Al(p,γ)26 Si reaction suffers from significant uncertainties due to the lack of relevant structure information in the compound nucleus 26 Si. An 25 Al+p elastic-scattering experiment in inverse kinematics was performed using the CRIB facility at the CNS at the Uni- versity of Tokyo, Japan, to try and improve current understanding. The 2 H(24 Mg,n)25 Al reaction was used to produce a 7.5 MeV/A 25 Al radioactive beam with intensities of ˜ 106 pps at the secondary CH2 target position. Protons were detected in silicon E?E telescopes and a center-of- mass energy range of 3 MeV was scanned, reaching up to about 8.5 MeV in excitation energy in 26 Si.

  4. 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.

  5. 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.

  6. Nitrogen deposition along differently exposed slopes in the Bavarian Alps.

    PubMed

    Kirchner, Manfred; Fegg, Wolfgang; Römmelt, Horst; Leuchner, Michael; Ries, Ludwig; Zimmermann, Ralf; Michalke, Bernd; Wallasch, Markus; Maguhn, Jürgen; Faus-Kessler, Theresa; Jakobi, Gert

    2014-02-01

    The Alps are affected by high nitrogen deposition, particularly in the fringe of the Northern and Southern Alps. In the framework of a two-year monitoring study performed in 2010 and 2011, we investigated the ammonia and nitrogen dioxide air concentration and ammonium and nitrate deposition at different altitudes between 700 and 1,600 ma.s.l. in the Garmisch-Partenkirchen district in the Upper Bavaria region (Germany). Four-weekly measurements of deposition collected with bulk open field samplers and under-crown were performed in a profile perpendicular to the axis of the Loisach valley; measurements were conducted at eight sites. Whereas open field deposition ranged from 5 to 11 kg ha(-1)a(-1), nitrogen throughfall has reached up to 21 kg ha(-1)a(-1). Data from the valley and the slopes were compared with measurements performed on the platform of the Environmental Research Station Schneefernerhaus (Zugspitze) at an altitude of 2,650 ma.s.l. For the rough estimation of the total yearly deposition rate of nitrogen, the canopy uptake model was applied. By regarding nitrogen uptake by the trees, total deposition can exceed the throughfall in all sites by up to 50%. Additionally, we estimated the total deposition from the sum of wet and dry deposition. On the one side, the wet deposition could be extrapolated from the open field deposition. On the other side, we used the inferential method to calculate the dry deposition on the basis of NH3 and NO2 air concentrations and their literature based deposition velocities. Since fixed deposition velocities are inappropriate particularly in complex orography, we tried to find correction factors based upon terrain characteristics and meteorological considerations. Temperature monitoring at the eight sites and wind measurements at two sites provided some evidence for the semi-empirical parameterization. Due to numerous imponderabilities, the results of the two methods were not consistent for all sites.

  7. Ambient Noise Tomography in the Eastern Alps of Europe

    NASA Astrophysics Data System (ADS)

    Behm, M.; Nakata, N.; Lloyd, S. M.; Bokelmann, G.

    2013-12-01

    ALPASS is an international passive seismic monitoring experiment aimed at understanding the upper mantle structure in the in the European Eastern Alps. Data were collected from May 2005 to June 2006 at about 50 locations with an average spacing of 20 km, and have been used for teleseismic travel time tomography and receiver function analysis in previous studies. We combine the ALPASS data from 23 broadband stations with additional data from the temporary CBP (Carpathian Basin Project) network (15 stations), and present results from ambient noise tomography applied to the region covering the easternmost part of the Alps and its transition to the adjacent tectonic provinces (Vienna Basin, Bohemian Massif, Dinarides). By turning each station into a virtual source, we are able to recover surface waves in the frequency range of 0.1 - 1.0 Hz, which are sensitive to depths of approximately 2 - 20 kilometres. The three-component recordings allow to distinguish between Rayleigh waves on the vertical/radial components and Love waves on the transverse component. On average, the Love waves have higher apparent velocity by about 10%. Owing to dense receiver spacing and high S/N ratio of the obtained interferograms, we are able to derive a large set of dispersion curves. The complicated 3D structure of the investigated region calls for a tomographic approach to transform these dispersion curves to be representative of local 1D structures. The results correlate well with surface geology and provide the input to inversion for the vertical shear-wave velocity distribution.

  8. 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).

  9. Miocene to Pleistocene Exhumation of the Southern European Alps

    NASA Astrophysics Data System (ADS)

    Reverman, R.; Fellin, M.; Herman, F.; Willett, S.

    2011-12-01

    We apply apatite fission-track, (U-Th-Sm)/He dating, 4He/3He diffusion profiles, and OSL thermochronology to construct a complete thermal history and constrain the exhumation and topographic evolution of the Adamello complex in the Southern European Alps. The Adamello complex is an Eocene-Oligocene pluton emplaced at a depth of 7 km, now exposed and dissected by deep valleys with a local relief of more than 2 km, which drain into larger fluvial systems overdeepened during the Messinian salinity crisis. Our aim is to determine which major erosional event during the Miocene-to-Pleistocene evolution of the Alps most affected the topographic development of the Adamello complex. We quantify exhumation rates using multiple methods: age-elevation relationships, estimates based on exhumation of a fossil partial retention zone, and cooling histories. We then compare these estimates with those predicted from a 3-D heat conduction model including topographic relief and erosion (Pecube; Braun, 2002) paired with the Neighborhood Algorithm, which is a two stage mathematical procedure that finds models, or parameters, which minimize the misfit to the data (Sambridge, 1999a,b). The predicted tectono-geomorphic history is tested for feasibility through comparison with known timing of tectonic and erosional events. Our apatite fission-track and (U-Th-Sm)/He results indicate that relief development alone cannot account for the episodic history and suggest a possible coupling between tectonics and erosion in the late Miocene, where the establishment of major drainages coincides with a period of increased rock uplift. The initial results from even lower temperature methods, 4He/3He diffusion profiles and OSL thermochronology, suggest an increase in exhumation rates in the Plio-Pleistocene, most likely associated with glacial reshaping.

  10. Carbonate dissolution and transport in H2O fluids during subduction revealed by diamond-bearing rocks from the Alps

    NASA Astrophysics Data System (ADS)

    Frezzotti, M.; Selverstone, J.; Sharp, Z. D.; Compagnoni, R.

    2011-12-01

    Here we discuss the fate of subducted carbonates and its implications for recycling of crustal carbon. Thermodynamic models predict little decarbonation along most subduction geotherms, and the mechanisms by which carbon is transferred from the subducting slab to the overlying mantle remain poorly constrained. Diamond-bearing fluid inclusions in garnet in oceanic metasedimentary rocks from Lago di Cignana (western Alps) represent the first occurrence of diamond from a low-temperature subduction complex of clearly oceanic origin (T ≤600°C; P ≥3.5 GPa). The presence of diamonds in and associated with fluid inclusions provides clear evidence of carbon transport by fluids at depths that are directly relevant to slab-mantle fluid transfer during subduction. At room temperature, the fluid inclusions contain aqueous fluid, a vapor bubble, and multiple solid daughter crystals. Daughter crystals identified by Raman spectroscopy and microprobe analysis include ubiquitous Mg-calcite/calcite and rutile, and less common diamond, quartz, paragonite, dawsonite, rhodochrosite, dypingite, and pentahydrite. Molecular CO2 is absent or in trace amounts. The aqueous liquid phase contains ≥0.2 wt%, HCO3-, CO32-, and SO42- ions. In Raman spectra, broad peaks at 773 and 1017 cm-1 point to the presence of both Si(OH)4(aq) and deprotonated monomers (e.g., SiO(OH)3-(aq), and SiO2(OH)22-(aq)), indicative of alkaline solutions. The absence of CO2 in the vapor, and the presence of carbonate daughter minerals, CO32-(aq), and HCO3-(aq) also show that the trapped fluids are alkaline at ambient conditions. High activities of aqueous carbon species reveal that carbonate dissolution is an important mechanism for mobilizing slab carbon at sub-arc depths (100-200 km) during oceanic subduction. Our results imply that the magnitude of carbon release and transport from the slab at sub-arc depths is greater than experimentally predicted on the basis of decarbonation reactions alone.

  11. Production of citric and oxalic acids and solubilization of calcium phosphate by Penicillium bilaii.

    PubMed Central

    Cunningham, J E; Kuiack, C

    1992-01-01

    An isolate of Penicillium bilaii previously reported to solubilize mineral phosphates and enhance plant uptake of phosphate was studied. Using agar media with calcium phosphate and the pH indicator alizarin red S, the influence of the medium composition on phosphate solubility and medium acidification was recorded. The major acidic metabolites produced by P. bilaii in a sucrose nitrate liquid medium were found to be oxalic acid and citric acid. Citric acid production was promoted under nitrogen-limited conditions, while oxalic acid production was promoted under carbon-limited conditions. Citric acid was produced in both growth and stationary phases, but oxalic acid production occurred only in stationary phase. When submerged cultures which normally produce acid were induced to sporulate, the culture medium shifted toward alkaline rather than acid reaction with growth. PMID:1622211

  12. 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.

  13. Exposure to the Proton Scavenger Glycine under Alkaline Conditions Induces Escherichia coli Viability Loss

    PubMed Central

    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. PMID:23544135

  14. Synthesis, mechanical and biological characterization of ionic doped carbonated hydroxyapatite/β-tricalcium phosphate mixtures.

    PubMed

    Kannan, S; Vieira, S I; Olhero, S M; Torres, P M C; Pina, S; da Cruz e Silva, O A B; Ferreira, J M F

    2011-04-01

    The influence of ionic substituents in calcium phosphates intended for bone and tooth replacement biomedical applications is an important research topic, owing to the essential roles played by trace elements in biological processes. The present study investigates the mechanical and biological evaluation of ionic doped hydroxyapatite/β-tricalcium phosphate mixtures which have been prepared by a simple aqueous precipitation method. Heat treating the resultant calcium phosphates in a carbonated atmosphere led to the formation of ionic doped carbonated hydroxyapatite/β-tricalcium phosphate mixtures containing the essential ions of biological apatite. The structural analysis determined by Rietveld refinement confirmed the presence of hydroxyapatite as the main phase, together with a considerable amount of β-tricalcium phosphate. Such phase assemblage is essentially due to the influence of substituted ions during synthesis. The results from mechanical tests proved that carbonate substitutions are detrimental for the mechanical properties of apatite-based ceramics. In vitro proliferation assays of osteoblastic-like cells (MC3T3-E1 cell line) to powders revealed that carbonate incorporation can either delay or accelerate MC3T3 proliferation, although reaching the same proliferation levels as control cells after 2 weeks in culture. Further, the powders enable pre-osteoblastic differentiation in a similar manner to control cells, as indirectly measured by ALP activity and Type-I collagen medium secretion. PMID:21146640

  15. AlpArray - Probing Alpine geodynamics with the next generation of geophysical experiments and techniques

    NASA Astrophysics Data System (ADS)

    Kissling, Edi; Hetenyi, György; AlpArray Working Group

    2014-05-01

    AlpArray is a European initiative to advance our understanding of orogenesis and its relationship to mantle and plate dynamics, surface processes, seismotectonics and seismic hazard in the Alps and the surrounding Apennines-Carpathians-Dinarides orogenic system. The initiative will integrate present-day Earth observables with high-resolution geophysical imaging of 3D structure and physical properties of the lithosphere and of the upper mantle, with focus on a high-end seismological array. With nearly three years of scientific and technical preparation, the start of AlpArray experiments is now on the horizon. In this presentation we overview the general idea and purpose of AlpArray, reason why the initiative focuses on the greater Alpine area and discuss some of the outstanding scientific questions. We provide an overview of the planned field efforts, mainly the overall AlpArray seismic network with 40 km average station spacing and a large aerial coverage. Further plans of targeted seismological experiments with regional interests as well as of other field measurement techniques (magnetotellurics, gravity) are also presented. The outline of collaborative projects between the numerous participating institutions and researchers are described in the frame of seven main AlpArray themes. These include state-of-the-art modelling techniques that will ultimately describe the dynamic evolution of the orogenic system in great detail. Finally, the most recent news regarding the organization of the AlpArray project will be summarized.

  16. UNIBRA - the German contribution to the AlpArray Seismic Network

    NASA Astrophysics Data System (ADS)

    Friederich, Wolfgang; Korn, Michael; Meier, Thomas; Plenefisch, Thomas; Ritter, Joachim; Rümpker, Georg; Thomas, Christine; Tilmann, Frederik; Wassermann, Joachim

    2016-04-01

    AlpArray is a European initiative to advance our understanding of orogenesis and its relationship to mantle dynamics, plate reorganizations, surface processes and seismic hazard in the Alps-Apennines-Carpathians-Dinarides orogenic system (cited from the AlpArray Science Plan). The core of the AlpArray project is a dense seismic network covering the entire Alpine region including its forelands. The network is composed of about 600 broadband seismic sensors of which 280 are operated permanently by national institutions and the remaining 320 are deployed temporarily by project partners for a time span of at least 2 years. In a joint effort, German universities contribute 71 temporary stations to the AlpArray Seismic Network, of which 55 are located in southern Germany and 16 in Austria. The instrumentation of the stations is heterogeneous and comprises Streckeisen, Nanometrics and Guralp sensors operated with EarthData, Nanometrics and Quanterra data loggers. The instruments are installed in remote areas and rarely used buildings with power supply. Some stations transfer data via mobile internet connection. The data will be collected at national EIDA centers which also serve as nodes for dissemination of data to the members of the AlpArray working group. Here, we describe installation procedures and setting of the German UNIBRA (UNIversity BRoadband Array) stations. In addition, we present noise characteristics and first records of teleseismic earthquakes. The descriptions may serve as a reference for later work with data from the AlpArray Seismic Network.

  17. Solubilization of insoluble phosphates by thermophilic fungi.

    PubMed

    Singh, C P; Mishra, M M; Yadav, K S

    1980-01-01

    The solubilization of tricalcium phosphate and rock phosphate and assimilation of solubilized P by thermophilic fungi isolated from compost were studied. The solubilization of tricalcium phosphate was greater than that of rock phosphate on inoculation with fungi in liquid medium, but growth of most of the fungi was greater in rock phosphate. Torula thermophila solubilized tricalcium phosphate maximally. There was solubilization of rock phosphate in semi-solid lignocellulose medium by Aspergillus fumigatus.

  18. 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.

  19. Modification of human placental alkaline phosphatase by periodate-oxidized 1,N6-ethenoadenosine monophosphate.

    PubMed Central

    Chang, G G; Shiao, M S; Lee, K R; Wu, J J

    1990-01-01

    Oxidation of 1,N6-ethenoadenosine monophosphate (epsilon AMP) with periodate cleaved the cis-diol of the ribose ring and resulted in the formation of a dialdehyde derivative (epsilon AMP-dial). At room temperature epsilon AMP-dial was unstable and underwent beta-elimination to give 4',5'-anhydro-1,N6-ethenoadenosine dialdehyde acetal (A epsilon Ado-dial). These nucleotide analogues were found to inactivate human placental alkaline phosphatase in a time- and concentration-dependent manner. epsilon AMP-dial was shown to be an affinity label for the enzyme on the basis of the following criteria. (a) Kinetics of the enzyme activity loss over a wide range of epsilon AMP-dial concentration showed a saturating phenomenon. Removal of the phosphate group made the reagent (A epsilon Ado-dial) become a general chemical modifying reagent. (b) The artificial substrate p-nitrophenyl phosphate gave substantial protection of the enzyme against inactivation. (c) epsilon AMP-dial was a substrate and a partial mixed-type inhibitor for the enzyme. Results of the inhibition and protection studies indicated that the reagent and substrate could combine with the enzyme simultaneously. Besides the phosphate-binding domain, an induced hydrophobic region is proposed for the substrate-binding site for human placental alkaline phosphatase. PMID:2176472

  20. Phosphorus Mobilization from Manure-Amended and Unamended Alkaline Soils to Overlying Water during Simulated Flooding.

    PubMed

    Amarawansha, E A G S; Kumaragamage, D; Flaten, D; Zvomuya, F; Tenuta, M

    2015-07-01

    Anaerobic soil conditions resulting from flooding often enhance release of phosphorus (P) to overlying water. Enhanced P release is well documented for flooded acidic soils; however, there is little information for flooded alkaline soils. We examined the effect of flooding and anaerobic conditions on P mobilization using 12 alkaline soils from Manitoba that were either unamended or amended with solid cattle manure. Pore water and floodwater were analyzed over 8 wk of simulated flooding for dissolved reactive P (DRP), Ca, Mg, Fe, and Mn. As expected, manured soils had significantly greater pore and floodwater DRP concentrations than unamended. Flooding increased pore water DRP concentrations significantly in all soils and treatments except one manured clay in which concentrations increased initially and then decreased. Floodwater DRP concentrations increased significantly by two- to 15-fold in 10 soils regardless of amendment treatment but remained relatively stable in the two soils with greatest clay content. Phosphorus release at the onset of flooding was associated with the release of Ca, Mg, and Mn, suggesting that P release may be controlled by the dissolution of Mg and Ca phosphates and reductive dissolution of Mn phosphates. Thereafter, P release was associated with release of Fe, suggesting the reductive dissolution of Fe phosphates. Differences in pore water and floodwater DRP concentrations among soils and amendment treatments and the high variability in P mobilization from pore water to floodwater among soils indicate the need to further investigate chemical reactions responsible for P release and mobility under anaerobic conditions. PMID:26437107

  1. Phosphorus Mobilization from Manure-Amended and Unamended Alkaline Soils to Overlying Water during Simulated Flooding.

    PubMed

    Amarawansha, E A G S; Kumaragamage, D; Flaten, D; Zvomuya, F; Tenuta, M

    2015-07-01

    Anaerobic soil conditions resulting from flooding often enhance release of phosphorus (P) to overlying water. Enhanced P release is well documented for flooded acidic soils; however, there is little information for flooded alkaline soils. We examined the effect of flooding and anaerobic conditions on P mobilization using 12 alkaline soils from Manitoba that were either unamended or amended with solid cattle manure. Pore water and floodwater were analyzed over 8 wk of simulated flooding for dissolved reactive P (DRP), Ca, Mg, Fe, and Mn. As expected, manured soils had significantly greater pore and floodwater DRP concentrations than unamended. Flooding increased pore water DRP concentrations significantly in all soils and treatments except one manured clay in which concentrations increased initially and then decreased. Floodwater DRP concentrations increased significantly by two- to 15-fold in 10 soils regardless of amendment treatment but remained relatively stable in the two soils with greatest clay content. Phosphorus release at the onset of flooding was associated with the release of Ca, Mg, and Mn, suggesting that P release may be controlled by the dissolution of Mg and Ca phosphates and reductive dissolution of Mn phosphates. Thereafter, P release was associated with release of Fe, suggesting the reductive dissolution of Fe phosphates. Differences in pore water and floodwater DRP concentrations among soils and amendment treatments and the high variability in P mobilization from pore water to floodwater among soils indicate the need to further investigate chemical reactions responsible for P release and mobility under anaerobic conditions.

  2. Preparation of acidic and alkaline macrocapsules for pH control.

    PubMed

    Flora, Joseph R V; Baker, Benjamin; Wybenga, Daniel; Zhu, Huiying; Aelion, C Marjorie

    2008-01-01

    A series of experiments was performed to prepare acidic macroencapsulated buffers composed of 20% Ca(H2PO4)(2) and 80% Eudragit S 100 polymer and alkaline macrocapsules composed of 65% K2HPO4 and 35% Eudragit E PO polymer (the powdered form of Eudragit E 100). Eudragit S 100 was shown to be soluble at a pH greater than 7.0, while Eudragit E 100 was soluble at a pH less than 7.0. Both polymers did not impart significant biochemical oxygen demand. The Eudragit E PO polymer solution showed low toxicity (EC50=91%) based on the Microtox Acute Toxicity Test compared to the 0.1mM background phosphate buffer solution (EC50=100%) while the Eudragit S 100 polymer solution showed higher toxicity (EC50=53%). Batch tests showed that the acidic macrocapsules reduced the pH of a 0.1mM phosphate solution from 11 to neutral, while the alkaline macrocapsules increased the pH of a 0.1mM phosphate solution from 3 to neutral. The macrocapsules could potentially be used as an in situ proportional pH controller for groundwater remediation.

  3. 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

  4. Late Oligocene to early Miocene lateral extrusion at the eastern border of the Lepontine dome of the central Alps (Bergell and Insubric areas, eastern central Alps)

    NASA Astrophysics Data System (ADS)

    Ciancaleoni, Laurent; Marquer, Didier

    2008-07-01

    How the internal domain of the central Alps deformed during late stages of the continental collision remains poorly understood. To fill this gap in knowledge, we present new data constraining the late Alpine brittle and brittle-ductile tectonics in Oligocene intrusions of the Bergell Alps (eastern central Alps). Syncollisional late normal faulting is widespread at all investigated scales, but part of the deformation observed is also associated with oblique-slip and transcurrent displacements. The faults are explained by stress permutations in space and time implying the three principal stress axes and correspond to different homoaxial states of stress of one single transtensive tectonic event. Motion on these faults accommodated coeval orogen-parallel extension and orogen-perpendicular contraction, during oblique indentation by the Southern alpine crust. Extensional and transcurrent structures likely formed by lateral extrusion of the Bergell Alps via distributed eastward extrusion of imbricated wedge-shaped crustal blocks, during the late Oligocene and lower Miocene. A first-order wedge-shaped crustal block in the Bergell Alps is represented by three coeval segments of the Periadriatic Fault System (namely the Tonale, Engadine, and Forcola faults). The eastern central segment of the Alpine belt appears to have deformed during the late Oligocene and lower Miocene in a similar way to its extremities where lateral escape tectonics occurred at least until the upper Miocene, according to free boundary availabilities.

  5. In vitro ALP and osteocalcin gene expression analysis and in vivo biocompatibility of N-methylene phosphonic chitosan nanofibers for bone regeneration.

    PubMed

    Datta, Pallab; Ghosh, Paulomi; Ghosh, Kuntal; Maity, Pritiprasanna; Samanta, Sintu Kumar; Ghosh, Sudip Kumar; Mohapatra, Pradeep Kumar Das; Chatterjee, Jyotirmoy; Dhara, Santanu

    2013-05-01

    Most polymeric nanofibers used for bone tissue engineering lack adequate functional groups for bioactivity. This study explores the potential of nanofibers of phosphate functionalized derivative of chitosan-N-methylene phosphonic chitosan (NMPC) for bone tissue engineering. Nanofibers were fabricated by electrospinning of NMPC/PVA blend solutions. NMPC/PVA nanofibers exhibited 172% higher viability of MG-63 cells compared to pure PVA nanofibers. ALP and Collagen type I genes revealed higher expression in NMPC nanofibers on day 3 whereas osteocalcin gene was expressed on day 7. In rabbit tibial defects, NMPC based electrospun graft showed presence of no adverse tissue reaction by histological examination while radiological examination suggested acceleration of bone healing by 300% compared to defects without any scaffold. Thus it is concluded NMPC based nanofibers may have potential for bone grafting applications. PMID:23802418

  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. Ecto-alkaline phosphatase activity identified at physiological pH range on intact P19 and HL-60 cells is induced by retinoic acid.

    PubMed

    Scheibe, R J; Kuehl, H; Krautwald, S; Meissner, J D; Mueller, W H

    2000-01-01

    The activity of membrane-bound alkaline phosphatase (ALP) expressed on the external surface of cultured murine P19 teratocarcinoma and human HL-60 myeloblastic leukemia cells was studied at physiological pH using p-nitrophenylphosphate (pNPP) as substrate. The rate of substrate hydrolysis catalyzed by intact viable cells remained constant for eight successive incubations of 30 min and was optimal at micromolar substrate concentrations over the pH range 7.4-8.5. The value of apparent K(m) for pNPP in P19 and HL-60 cells was 120 microM. Hydrolytic activity of the ecto-enzyme at physiological pH decreased by the addition of levamisole, a specific and noncompetitive inhibitor of ALP (K(i) P19 = 57 microM; K(i) HL-60 = 50 microM). Inhibition of hydrolysis was reversed by removal of levamisole within 30 min. Retinoic acid (RA), which promotes the differentiation of P19 and HL-60 cells, induced levamisole-sensitive ecto-phosphohydrolase activity at pH 7.4. After its autophosphorylation by ecto-kinase activity, a 98-kDa membrane protein in P19 cells was found to be sensitive to ecto-ALP, and protein dephosphorylation increased after incubation of cells with RA for 24 h and 48 h. Orthovanadate, an inhibitor of all phosphatase activities, blocked the levamisole-sensitive dephosphorylation of the membrane phosphoproteins, while (R)-(-)-epinephrine reversed the effect by complexation of the inhibitor. The results demonstrate that the levamisole-sensitive phosphohydrolase activity on the cell surface is consistent with ecto-ALP activity degrading both physiological concentrations of exogenously added substrate and endogenous surface phosphoproteins under physiological pH conditions. The dephosphorylating properties of ecto-ALP are induced by RA, suggesting a specific function in differentiating P19 teratocarcinoma and HL-60 myeloblastic leukemia cells. PMID:10649440

  8. A highly sensitive homogeneous electrochemical assay for alkaline phosphatase activity based on single molecular beacon-initiated T7 exonuclease-mediated signal amplification.

    PubMed

    Zhang, Lianfang; Hou, Ting; Li, Haiyin; Li, Feng

    2015-06-21

    Alkaline phosphatase (ALP), a class of enzymes that catalyzes the dephosphorylation of a variety of substrates, is one of the most commonly assayed enzymes in routine clinical practice, and an important biomarker related to many human diseases. Herein, a facile and highly sensitive homogeneous electrochemical biosensing strategy was proposed for the ALP activity detection based on single molecular beacon-initiated T7 exonuclease-assisted signal amplification. One 3'-phosphorylated and 5'-methylene blue (MB) labeled hairpin probe (HP) is ingeniously designed. In the presence of ALP, the dephosphorylation of HP, the subsequent Klenow fragment (KF) polymerase-catalyzed elongation and T7 exonuclease-catalyzed digestion of the duplex stem of HP take place, releasing MB-labeled mononucleotides and the trigger DNA (tDNA). tDNA then hybridizes with another HP and initiates the subsequent cycling cleavage process. As a result, a large amount of MB-labeled mononucleotides are released, generating a significantly amplified electrochemical signal toward the ALP activity assay. A directly measured detection limit as low as 0.1 U L(-1) is obtained, which is comparable to that of the fluorescence method and up to three orders of magnitude lower than that of the immobilization-based electrochemical strategy previously reported. In addition to high sensitivity and good selectivity, the as-proposed strategy also exhibits the advantages of simplicity and convenience, because the assay is carried out in the homogeneous solution phase and sophisticated electrode modification processes are avoided. Therefore, the homogeneous electrochemical method we proposed here is an ideal candidate for ALP activity detection in biochemical research and clinical practices. PMID:25924941

  9. A highly sensitive homogeneous electrochemical assay for alkaline phosphatase activity based on single molecular beacon-initiated T7 exonuclease-mediated signal amplification.

    PubMed

    Zhang, Lianfang; Hou, Ting; Li, Haiyin; Li, Feng

    2015-06-21

    Alkaline phosphatase (ALP), a class of enzymes that catalyzes the dephosphorylation of a variety of substrates, is one of the most commonly assayed enzymes in routine clinical practice, and an important biomarker related to many human diseases. Herein, a facile and highly sensitive homogeneous electrochemical biosensing strategy was proposed for the ALP activity detection based on single molecular beacon-initiated T7 exonuclease-assisted signal amplification. One 3'-phosphorylated and 5'-methylene blue (MB) labeled hairpin probe (HP) is ingeniously designed. In the presence of ALP, the dephosphorylation of HP, the subsequent Klenow fragment (KF) polymerase-catalyzed elongation and T7 exonuclease-catalyzed digestion of the duplex stem of HP take place, releasing MB-labeled mononucleotides and the trigger DNA (tDNA). tDNA then hybridizes with another HP and initiates the subsequent cycling cleavage process. As a result, a large amount of MB-labeled mononucleotides are released, generating a significantly amplified electrochemical signal toward the ALP activity assay. A directly measured detection limit as low as 0.1 U L(-1) is obtained, which is comparable to that of the fluorescence method and up to three orders of magnitude lower than that of the immobilization-based electrochemical strategy previously reported. In addition to high sensitivity and good selectivity, the as-proposed strategy also exhibits the advantages of simplicity and convenience, because the assay is carried out in the homogeneous solution phase and sophisticated electrode modification processes are avoided. Therefore, the homogeneous electrochemical method we proposed here is an ideal candidate for ALP activity detection in biochemical research and clinical practices.

  10. 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. PMID:26561448

  11. 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.

  12. 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.

  13. Inorganic-organic separators for alkaline batteries

    NASA Technical Reports Server (NTRS)

    Sheibley, D. W. (Inventor)

    1978-01-01

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

  14. 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.

  15. 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.

  16. Dehydrogenases, Acid and Alkaline Phosphatases, and Esterases for Chemotaxonomy of Selected Meloidogyne, Ditylenchus, Heterodera and Aphelenchus spp.

    PubMed Central

    Dickson, D. W.; Huisingh, D.; Sasser, J. N.

    1971-01-01

    Various taxonomically useful profiles of four dehydrogenases (lactate, malate, glucose-6-phosphate, and a-glycerophosphate) and three hydrolases (acid and alkaline phosphatase and esterase) were detected in whole nematode homogenates of Meloidogyne javanica, M. hapla, M. incognita, M. arenaria, Ditylenchus dipsaci, D. triformis, Heterodera glycines, and Aphelenchus avenae. The enzyme profiles were stable in populations cultured on several different hosts. A tentative enzymically-determined phylogeny of Meloidogyne is given. PMID:19322334

  17. Spatial and temporal patterns of wildfires in the Northern Alps

    NASA Astrophysics Data System (ADS)

    Heel, Michael; Sass, Oliver; Friedmann, Arne; Wetzel, Karl-Friedrich

    2010-05-01

    Wildfires in the northern Alps are rare compared to e.g. the Mediterranean region. However, fires occurring on the dry, south-exposed slopes of the inner-alpine valleys can constitute a significant disturbance of the ecosystems in the sub-alpine belt. We reconstructed the younger regional wildfire history (last few centuries) of a part of the the Northern Limestone Alps using chronicles, forestry and fire brigade records as well as historical pictures (postcards, aerial photos etc.), local names and interviews with local people. The long-term fire frequency was investigated using mire drillings, charcoal in soils and dendrochronology. In the surrounding of the Karwendel, Wetterstein and Mieminger Mountains we have identified c. 400 forest fires to date. The earliest detected fire dates to more than 2900 years; the largest one (in 1705) affected an area of several thousand hectares. Approximately 90% of the fires are man-made (negligence, arson, railway) which explains the concentration on the south-exposed slopes of the densely populated Inn valley. Most of the larger fires take place in the altitudinal belt between 1400 and 1900 m a.s.l.; apart from very few exceptions, they are restricted to southerly orientations. Locally, mean recurrence intervals of 200-300 years occur which is similar to e.g. boreal forests in Canada. We observed a strong seasonality with 40% of the fires occurring in spring and 30% in summer. There is a weak correlation with the weather conditions in the one or two weeks before the fire with dry periods promoting wildfire ignition and burnt area size; however, there are many exceptions from the rule. The 1940ies stands out for more than twice as much fires than in all other decades which is both due to climatic and anthropogenic causes. Today, there is an apparent trend towards more frequent and smaller fires. The frequency is biased by the multitude of available documentation today (e.g. websites of fire brigades), while the decreasing size

  18. 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

  19. Prograde and retrograde thermal histories from the central Swiss Alps

    NASA Astrophysics Data System (ADS)

    Vance, Derek; Keith O'Nions, R.

    1992-12-01

    The "Lepontine" phase of metamorphism in the central Alps is a result of thermal relaxation following continental collision and overthrusting. The timing of the peak remains poorly known, largely because the existing chronological data relate to cooling or to phases for which the pressure and temperature of growth is only poorly constrained. Here we report U sbnd Pb, Sm sbnd Nd and Rb sbnd Sr analyses of garnets from the central Swiss Alps, as well as major element data on garnets and other phases. The data shows that the climax of the metamorphism was diachronous across grade, from 30 Ma in the lower grade northwest to 27 Ma in the higher grade in the southeast. Consideration of these data in conjunction with existing chronological constraints suggests a period of about 10 Ma between the attainment of the metamorphic climax in greenschist facies and the upper amphibolite facies rocks. The time constraints obtained here yield a garnet growth interval of 2.9 ± 1.5 Ma, corresponding to a growth rate of 0.2 cm Ma -1, a heating rate of 12°C Ma -1 and a burial rate of 2.9 km Ma -1. The strain rate obtained from measurement of the rotation of the garnet relative to the matrix fabric during growth is ˜ 1.9 × 10 -14s -1. This strain rate, as well as the heating and burial rates, are in the range of values derived in a similar fashion from other Barrovian metamorphic terrains. Consideration of these data in conjunction with published cooling ages leads to cooling rates of about 20°C Ma -1 for the Steinental samples but much higher rates for the Bellinzona area (> 100°C Ma -1). In both cases it is necessary to invoke tectonic exhumation rather than isostatic recovery and erosion to explain the observed temperature-time trends. In the case of the Bellinzona area, the most likely cause of the very rapid cooling rates is late backthrusting over the Insubric Line.

  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

    2012-06-01

    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. PMID:22571620

  1. 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

    2012-06-01

    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.

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

    PubMed

    Park, Jung-Hui; Lee, Eun-Jung; Knowles, Jonathan C; Kim, Hae-Won

    2014-03-01

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

  3. Separating and Stabilizing Phosphate from High-Level Radioactive Waste: Process Development and Spectroscopic Monitoring

    SciTech Connect

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

    2012-05-09

    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.

  4. Determination of direct available phosphate in fertilizers by a discrete analyzer: single-laboratory validation.

    PubMed

    Nimick, Ryan; Mehlbrech, Laura; Novotny, Lawrence; Thiex, Nancy

    2012-01-01

    To improve throughput during peak seasonal demand, a screening method for the determination of fertilizer-available phosphate using a discrete analyzer for semi-automation was validated in a single laboratory. The fertilizer materials were extracted using a neutral EDTA-ammonium citrate solution as detailed in AOAC Official Method 993.31. Phosphate was subsequently freed from the matrix and converted to orthophosphate using an alkaline persulfate digestion modified from a U.S. Geological Survey water method. Phosphorus was determined colorimetrically on a discrete analyzer. Twelve check samples from the Magruder Fertilizer Check Sample Program and Association of Fertilizer and Phosphate Chemists Fertilizer Check Program were used for method validation experiments. The proposed method is linear from 0.01 to 20 mg/L (ppm) phosphorus. Recovery for all materials averaged 101%, with a range of 99.2 to 103%. Bias for all materials averaged 0.59% with a range of -0.11 to 1.68%, with bias increasing at concentrations of available phosphate exceeding 40%. The LOD was calculated to be 0.001% available phosphate and the LOQ 0.002% available phosphate. The method was found fit for purpose as a screening method for available phosphate analysis in fertilizers.

  5. Sustained release poly (lactic-co-glycolic acid) microspheres of bone morphogenetic protein 2 plasmid/calcium phosphate to promote in vitro bone formation and in vivo ectopic osteogenesis

    PubMed Central

    Qiao, Chunyan; Zhang, Kai; Sun, Bin; Liu, Jinzhong; Song, Jiyu; Hu, Yue; Yang, Shihui; Sun, Hongchen; Yang, Bai

    2015-01-01

    Bone regeneration often requires continuous stimulation to promote local bone formation. In the present study, calcium phosphate (CaPi) was used to promote transfection of human bone morphogenetic protein 2 (BMP-2) cDNA plasmid, and poly (lactic-co-glycolic acid) (PLGA) was used to prepare microspheres of pBMP-2/CaPi (i.e., PLGA@pBMP-2/CaPi) using W/O/W double emulsion solvent evaporation method. We showed that PLGA@pBMP-2/CaPi microspheres were spherical with smooth surface, and the particle size ranged from 0.5 to 35 μm. Encapsulation efficiency was up to 30~50%. The release of BMP-2 cDNA from microspheres continued more than 30 days and constituted, less than 7.5% of total plasmid amount within the first 24 h. Real-time PCR results showed that co-culturing of PLGA@pBMP-2/CaPi with bone marrow-derived mesenchymal stem cells (BMSCs) increased calcium deposition and gene expressions of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), SP7, and collagen type I (COLL I) in a time-dependent manner. Finally, X-ray analysis demonstrated that in vivo delivery of PLGA@pBMP-2/CaPi microspheres into the tibialis anterior muscles of rats promoted the generation of osteoblasts, bone tissue, and bone structure. The findings suggested that PLGA@pBMP-2/CaPi microspheres can promote ectopic osteogenesis in non-bone tissues, with strong prospects in promoting bone regeneration. PMID:26885257

  6. 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.

  7. 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.

  8. The Arabidopsis alkaline ceramidase TOD1 is a key turgor pressure regulator in plant cells.

    PubMed

    Chen, Li-Yu; Shi, Dong-Qiao; Zhang, Wen-Juan; Tang, Zuo-Shun; Liu, Jie; Yang, Wei-Cai

    2015-01-01

    Turgor pressure plays pivotal roles in the growth and movement of walled cells that make up plants and fungi. However, the molecular mechanisms regulating turgor pressure and the coordination between turgor pressure and cell wall remodelling for cell growth remain poorly understood. Here, we report the characterization of Arabidopsis TurgOr regulation Defect 1 (TOD1), which is preferentially expressed in pollen tubes and silique guard cells. We demonstrate that TOD1 is a Golgi-localized alkaline ceramidase. tod1 mutant pollen tubes have higher turgor than wild type and show growth retardation both in pistils and in agarose medium. In addition, tod1 guard cells are insensitive to abscisic acid (ABA)-induced stomatal closure, whereas sphingosine-1-phosphate, a putative downstream component of ABA signalling and product of alkaline ceramidases, promotes closure in both wild type and tod1. Our data suggest that TOD1 acts in turgor pressure regulation in both guard cells and pollen tubes.

  9. ALP conversion and the soft X-ray excess in the outskirts of the Coma cluster

    SciTech Connect

    Kraljic, David; Rummel, Markus; Conlon, Joseph P. E-mail: Markus.Rummel@physics.ox.ac.uk

    2015-01-01

    It was recently found that the soft X-ray excess in the center of the Coma cluster can be fitted by conversion of axion-like-particles (ALPs) of a cosmic axion background (CAB) to photons. We extend this analysis to the outskirts of Coma, including regions up to 5 Mpc from the center of the cluster. We extract the excess soft X-ray flux from ROSAT All-Sky Survey data and compare it to the expected flux from ALP to photon conversion of a CAB. The soft X-ray excess both in the center and the outskirts of Coma can be simultaneously fitted by ALP to photon conversion of a CAB. Given the uncertainties of the cluster magnetic field in the outskirts we constrain the parameter space of the CAB. In particular, an upper limit on the CAB mean energy and a range of allowed ALP-photon couplings are derived.

  10. Inositol phosphates in the environment.

    PubMed Central

    Turner, Benjamin L; Papházy, Michael J; Haygarth, Philip M; McKelvie, Ian D

    2002-01-01

    The inositol phosphates are a group of organic phosphorus compounds found widely in the natural environment, but that represent the greatest gap in our understanding of the global phosphorus cycle. They exist as inositols in various states of phosphorylation (bound to between one and six phosphate groups) and isomeric forms (e.g. myo, D-chiro, scyllo, neo), although myo-inositol hexakisphosphate is by far the most prevalent form in nature. In terrestrial environments, inositol phosphates are principally derived from plants and accumulate in soils to become the dominant class of organic phosphorus compounds. Inositol phosphates are also present in large amounts in aquatic environments, where they may contribute to eutrophication. Despite the prevalence of inositol phosphates in the environment, their cycling, mobility and bioavailability are poorly understood. This is largely related to analytical difficulties associated with the extraction, separation and detection of inositol phosphates in environmental samples. This review summarizes the current knowledge of inositol phosphates in the environment and the analytical techniques currently available for their detection in environmental samples. Recent advances in technology, such as the development of suitable chromatographic and capillary electrophoresis separation techniques, should help to elucidate some of the more pertinent questions regarding inositol phosphates in the natural environment. PMID:12028785

  11. Clinoenstatite in alpe arami peridotite: additional evidence of very high pressure

    PubMed

    Bozhilov; Green; Dobrzhinetskaya

    1999-04-01

    Observations by transmission electron microscopy show that lamellae of clinoenstatite are present in diopside grains of the Alpe Arami garnet lherzolite of the Swiss Alps. The simplest interpretation of the orientation, crystallography, and microstructures of the lamellae and the phase relationships in this system is that the lamellae originally exsolved as the high-pressure C-centered form of clinoenstatite. These results imply that the rocks were exhumed from a minimum depth of 250 kilometers before or during continental collision. PMID:10102809

  12. Clinoenstatite in alpe arami peridotite: additional evidence of very high pressure

    PubMed

    Bozhilov; Green; Dobrzhinetskaya

    1999-04-01

    Observations by transmission electron microscopy show that lamellae of clinoenstatite are present in diopside grains of the Alpe Arami garnet lherzolite of the Swiss Alps. The simplest interpretation of the orientation, crystallography, and microstructures of the lamellae and the phase relationships in this system is that the lamellae originally exsolved as the high-pressure C-centered form of clinoenstatite. These results imply that the rocks were exhumed from a minimum depth of 250 kilometers before or during continental collision.

  13. Composite seal reduces alkaline battery leakage

    NASA Technical Reports Server (NTRS)

    Clatterbuck, C. H.; Plitt, K. F.

    1965-01-01

    Composite seal consisting of rubber or plastic washers and a metal washer reduces alkaline battery leakage. Adhesive is applied to each washer interface, and the washers are held together mechanically.

  14. CLIM-RUN: Tourism cas study over the French Alps

    NASA Astrophysics Data System (ADS)

    Dubois, C.

    2012-04-01

    Climate information for societal use has becoming a major challenge for tourism management and adaptation in a context of strong climate variability and change. Within the CLIMRUN EU FP7, a case study on summer tourism in the French Alps has been identify. I will introduce the bottom-up approach use in the project where stakeholders and local users meet with climate experts. From those meetings, they thus identify the climate dependence and information which impact their summer activities over this region. All the activities are located in a mountainous region where outdoor leisure is the main economic driver of the region. It has emerged that the climate requirements are as well on past as on future climate information. On one side, the past climate parameters are found to be an invaluable information to evaluate the climate dependence of the different activities. A better knowledge as well as a growing interest in climate variability has been express to quantify the climate dependence on their activities. On the other side, the future climate information requested mainly on seasonal to decadal timescale. A particular interest has been express on the snow cover at the end of the winter season, evolution of heavy precipitations, heatwave, air temperatures and well as the water temperature of the mountainous lakes. Those climate variables are used to create comfort index under climate change. All those targeted climate information are based on on-going projects as well as future model development.

  15. Negative elevation-dependent warming trend in the Eastern Alps

    NASA Astrophysics Data System (ADS)

    Tudoroiu, M.; Eccel, E.; Gioli, B.; Gianelle, D.; Schume, H.; Genesio, L.; Miglietta, F.

    2016-04-01

    Mountain regions and the important ecosystem services they provide are considered to be very vulnerable to the current warming, and recent studies suggest that high-mountain environments experience more rapid changes in temperature than environments at lower elevations. Here we analysed weather records for the period 1975-2010 from the Eastern Italian Alps that show that warming occurred both at high and low elevations, but it was less pronounced at high elevations. This negative elevation-dependent trend was consistent for mean, maximum and minimum air temperature. Global radiation data measured at different elevations, surface energy fluxes measured above an alpine grassland and above a coniferous forest located at comparable elevations for nine consecutive years as well as remote sensing data (MODIS) for cloud cover and aerosol optical depth were analysed to interpret this observation. Increasing global radiation at low elevations turned out to be a potential driver of this negative elevation-dependent warming, but also contributions from land use and land cover changes at high elevations (abandonment of alpine pastures, expansion of secondary forest succession) were taken into account. We emphasise though, that a negative elevation-dependent warming is not universal and that future research and in particular models should not neglect the role of land use changes when determining warming rates over elevation.

  16. Advanced Large Area Plastic Scintillator Project (ALPS): Final Report

    SciTech Connect

    Jordan, David V.; Reeder, Paul L.; Todd, Lindsay C.; Warren, Glen A.; McCormick, Kathleen R.; Stephens, Daniel L.; Geelhood, Bruce D.; Alzheimer, James M.; Crowell, Shannon L.; Sliger, William A.

    2008-02-05

    The advanced Large-Area Plastic Scintillator (ALPS) Project at Pacific Northwest National Laboratory investigated possible technological avenues for substantially advancing the state-of-the-art in gamma-ray detection via large-area plastic scintillators. The three predominant themes of these investigations comprised the following: * Maximizing light collection efficiency from a single large-area sheet of plastic scintillator, and optimizing hardware event trigger definition to retain detection efficiency while exploiting the power of coincidence to suppress single-PMT "dark current" background; * Utilizing anti-Compton vetoing and supplementary spectral information from a co-located secondary, or "Back" detector, to both (1) minimize Compton background in the low-energy portion of the "Front" scintillator's pulse-height spectrum, and (2) sharpen the statistical accuracy of the front detector's low-energy response prediction as impelmented in suitable energy-windowing algorithms; and * Investigating alternative materials to enhance the intrinsic gamma-ray detection efficiency of plastic-based sensors.

  17. Snow survey and vegetation growth in high mountains (Swiss Alps)

    NASA Technical Reports Server (NTRS)

    Haefner, H. (Principal Investigator)

    1973-01-01

    The author has identified the following significant results. A method for mapping snow over large areas was developed combining the possibilities of a Quantimet (QTM 72) to evaluate the exact density level of the snow cover for each individual image (or a selected section of the photo) with the higher resolution of photographic techniques. The density level established on the monitor by visual control is used as reference for the exposure time of a lithographic film, producing a clear tonal separation of all snow- and ice-covered areas from uncovered land in black and white. The data is projected onto special maps 1:500,000 or 1:100,000 showing the contour lines and the hydrographic features only. The areal extent of the snow cover may be calculated directly with the QTM 720 or on the map. Bands 4 and 5 provide the most accurate results for mapping snow. Using all four bands a separation of an old melting snow cover from a new one is possible. Regional meteorological studies combining ERTS-1 imagery and conventional sources describe synoptical evolution of meteorological systems over the Alps.

  18. Thrusting and wedge growth, Southern Alps of Lombardia (Italy)

    NASA Astrophysics Data System (ADS)

    Roeder, Dietrich

    1992-06-01

    A south-vergent fold-thrust belt of Miocene-Recent age accompanies the south slope of the Lombardian Alps and is partly buried beneath Plio-Pleistocene Po Valley basin fill. The belt is probably detached along a trans-crustal thrust, named Main South Alpine Thrust (MSAT), with an estimated dip slip of 70-100 km. Transport on this thrust piggybacks the Adamello pluton of Late Eocene age, pre-Adamello folds, and Oligocene-Miocene Insubric strike-slip structures, by ramping up through 12-15 km of Austro-Alpine (Adria) crust and through 8-10 km of Triassic to Eocene sediments. Folds in the Front Ranges are ascribed to MSAT ramping, not to pre-Adamello compression. The MSAT soles upward in a blind thrust beneath 3-4 km of Oligocene-Pliocene foredeep fill. Initial regional failure along the MSAT implies substantial and pre-existing topographic relief near the Insubric line. An average of 25% wedge thickening during MSAT transport is consistent with the requirement of Coulomb critical taper. Progression of the south-Alpine detachment from the MSAT to the base of the foreland sediments has added a thickness of 6-12 km in footwall imbrications to the base and the toe of the thrust wedge. This addition in wedge volume is consistent with wedge dynamics only if a mid-Miocene or younger spike of excess Alpine topography is admitted.

  19. The Permian and Triassic in the Albanian Alps

    NASA Astrophysics Data System (ADS)

    Gaetani, Maurizio; Meço, Selam; Rettori, Roberto; Henderson, Charles M.; Tulone, Accursio

    2015-09-01

    The sedimentary succession of the Permian to Middle Triassic of the Albanian Alps is described, as part of the eastern Adria passive margin towards the Tethys. A carbonate ramp deepening towards NE in present day coordinates developed during the Middle Permian and was affected by block faulting with the deposition of carbonate breccia. The Early Triassic was characterized by intense terrigenous deposition with several cobble conglomerate units up to 80 m-thick, and by oolitic carbonate shoals. The fine clastic deposition ended gradually during the earliest Anisian and a wide calcarenitic ramp occupied the area, with small local carbonate mounds. Basinward, the red nodular limestone of the Han Bulog Formation was interbedded with calcarenitic material exported from the ramp. Drowning to more open conditions occurred towards the end of the Pelsonian. Subsequently, cherty limestone and tuffitic layers spread over the entire area. Towards the end of the Ladinian, with the end of the volcanic activity, red pelagic limestone was deposited locally for a short period. By the latest Ladinian most of the area returned to shallow-water conditions, with a peritidal carbonate platform. In the Theth area, in contrast, a basin with black organic-rich dolostone and limestone developed which seems to be unique in that part of the Adria passive margin. The occurrence of cobble conglomerate units in the Lower Triassic testifies to very active block faulting and high accommodation, not yet described for the area.

  20. Evaluation of the alkaline electrolysis of zinc

    SciTech Connect

    Meisenhelder, J.H.; Brown, A.P.; Loutfy, R.O.; Yao, N.P.

    1981-05-01

    The alkaline leach and electrolysis process for zinc production is compared to the conventional acid-sulfate process in terms of both energy saving and technical merit. In addition, the potential for industrial application of the alkaline process is discussed on the basis of present market conditions, possible future zinc market scenarios, and the probability of increased secondary zinc recovery. In primary zinc production, the energy-saving potential for the alkaline process was estimated to be greater than 10%, even when significantly larger electrolysis current densities than those required for the sulfate process are used. The principal technical advantages of the alkaline process are that it can handle low-grade, high-iron-content or oxidized ores (like most of those found in the US) in a more cost- and energy-efficient manner than can the sulfate process. Additionally, in the electrowinning operation, the alkaline process should be technically superior because a dendritic or sponge deposit is formed that is amenable to automated collection without interruption of the electrolysis. Also, use of the higher current densities would result in significant capital cost reductions. Alkaline-based electrolytic recovery processes were considered for the recycling of zinc from smelter baghouse dusts and from the potential source of nickel/zinc electric-vehicle batteries. In all comparisons, an alkaline process was shown to be technically superior and, particularly for the baghouse dusts, energetically and economically superior to alternatively proposed recovery methods based on sulfate electrolysis. It is concluded that the alkaline zinc method is an important alternative technology to the conventional acid zinc process. (WHK)

  1. 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.

  2. 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.

  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. Light weight phosphate cements

    DOEpatents

    Wagh, Arun S.; Natarajan, Ramkumar,; Kahn, David

    2010-03-09

    A sealant having a specific gravity in the range of from about 0.7 to about 1.6 for heavy oil and/or coal bed methane fields is disclosed. The sealant has a binder including an oxide or hydroxide of Al or of Fe and a phosphoric acid solution. The binder may have MgO or an oxide of Fe and/or an acid phosphate. The binder is present from about 20 to about 50% by weight of the sealant with a lightweight additive present in the range of from about 1 to about 10% by weight of said sealant, a filler, and water sufficient to provide chemically bound water present in the range of from about 9 to about 36% by weight of the sealant when set. A porous ceramic is also disclosed.

  5. Templated, layered manganese phosphate

    DOEpatents

    Thoma, Steven G.; Bonhomme, Francois R.

    2004-08-17

    A new crystalline maganese phosphate composition having an empirical formula: O). The compound was determined to crystallize in the trigonal space group P-3c1 with a=8.8706(4) .ANG., c=26.1580(2) .ANG., and V (volume)=1783 .ANG..sup.3. The structure consists of sheets of corner sharing Mn(II)O.sub.4 and PO.sub.4 tetrahedra with layers of (H.sub.3 NCH.sub.2 CH.sub.2).sub.3 N and water molecules in-between. The pronated (H.sub.3 NCH.sub.2 CH.sub.2).sub.3 N molecules provide charge balancing for the inorganic sheets. A network of hydrogen bonds between water molecules and the inorganic sheets holds the structure together.

  6. Comparison of phosphate materials for immobilizing cadmium in soil.

    PubMed

    Hong, Chang Oh; Chung, Doug Young; Lee, Do Kyoung; Kim, Pil Joo

    2010-02-01

    A study was conducted to compare the effects of phosphate (P) materials in reducing cadmium extractability. Seven P materials (commercial P fertilizers--fused phosphate (FP), 'fused and superphosphate' [FSP], and rock phosphate [RP]; P chemicals--Ca[H(2)PO(4)](2).H(2)O, [NH(4)](2)HPO(4), KH(2)PO(4), and K(2)HPO(4)) were selected for the test. The selected P source was mixed with Cd-contaminated soil at the rate of 0, 200, 400, 800, and 1,600 mg P kg(-1) under controlled moisture conditions at 70% of water holding capacity, then incubated for 8 weeks. FP, Ca(H(2)PO(4))(2) H(2)O, KH(2)PO(4), and K(2)HPO(4) significantly decreased NH(4)OAc-extractable Cd (plant-available form) concentrations with increasing application rates. Compared to other phosphate materials used, K(2)HPO(4) was found to be the most effective in reducing the plant-available Cd concentration in soil, mainly due to the negative charge increase caused by soil pH and phosphate adsorption. Contrary to the general information, FSP and (NH(4))(2)HPO(4) increased Cd extractability at low levels of P application (<400 mg kg(-1)), and thereafter Cd extractability decreased significantly with increasing application rate. RP scarcely had an effect on reducing Cd extractability. Ion activity products of CdHPO(4), Cd(OH)(2), and CdCO(3) analyzed by the MINTEQ program were significantly increased by K(2)HPO(4) addition, but the effect of Cd-P compound formation on reducing Cd extractability was negligible. Conclusively, the P-induced alleviation of Cd extractability can be attributed primarily to Cd immobilization due to the increase in soil pH and negative charge rather than Cd-P precipitation, and therefore, alkaline P materials such as K(2)HPO(4) are effective for immobilizing soil Cd. PMID:19633979

  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. Phosphate nutrition: improving low-phosphate tolerance in crops.

    PubMed

    López-Arredondo, Damar Lizbeth; Leyva-González, Marco Antonio; González-Morales, Sandra Isabel; López-Bucio, José; Herrera-Estrella, Luis

    2014-01-01

    Phosphorus is an essential nutrient that is required for all major developmental processes and reproduction in plants. It is also a major constituent of the fertilizers required to sustain high-yield agriculture. Levels of phosphate--the only form of phosphorus that can be assimilated by plants--are suboptimal in most natural and agricultural ecosystems, and when phosphate is applied as fertilizer in soils, it is rapidly immobilized owing to fixation and microbial activity. Thus, cultivated plants use only approximately 20-30% of the applied phosphate, and the rest is lost, eventually causing water eutrophication. Recent advances in the understanding of mechanisms by which wild and cultivated species adapt to low-phosphate stress and the implementation of alternative bacterial pathways for phosphorus metabolism have started to allow the design of more effective breeding and genetic engineering strategies to produce highly phosphate-efficient crops, optimize fertilizer use, and reach agricultural sustainability with a lower environmental cost. In this review, we outline the current advances in research on the complex network of plant responses to low-phosphorus stress and discuss some strategies used to manipulate genes involved in phosphate uptake, remobilization, and metabolism to develop low-phosphate-tolerant crops, which could help in designing more efficient crops.

  10. Site-directed mutagenesis maps interactions that enhance cognate and limit promiscuous catalysis by an alkaline phosphatase superfamily phosphodiesterase.

    PubMed

    Wiersma-Koch, Helen; Sunden, Fanny; Herschlag, Daniel

    2013-12-23

    Catalytic promiscuity, an evolutionary concept, also provides a powerful tool for gaining mechanistic insights into enzymatic reactions. Members of the alkaline phosphatase (AP) superfamily are highly amenable to such investigation, with several members having been shown to exhibit promiscuous activity for the cognate reactions of other superfamily members. Previous work has shown that nucleotide pyrophosphatase/phosphodiesterase (NPP) exhibits a >10⁶-fold preference for the hydrolysis of phosphate diesters over phosphate monoesters, and that the reaction specificity is reduced 10³-fold when the size of the substituent on the transferred phosphoryl group of phosphate diester substrates is reduced to a methyl group. Here we show additional specificity contributions from the binding pocket for this substituent (herein termed the R' substituent) that account for an additional ~250-fold differential specificity with the minimal methyl substituent. Removal of four hydrophobic side chains suggested on the basis of structural inspection to interact favorably with R' substituents decreases phosphate diester reactivity 10⁴-fold with an optimal diester substrate (R' = 5'-deoxythymidine) and 50-fold with a minimal diester substrate (R' = CH₃). These mutations also enhance the enzyme's promiscuous phosphate monoesterase activity by nearly an order of magnitude, an effect that is traced by mutation to the reduction of unfavorable interactions with the two residues closest to the nonbridging phosphoryl oxygen atoms. The quadruple R' pocket mutant exhibits the same activity toward phosphate diester and phosphate monoester substrates that have identical leaving groups, with substantial rate enhancements of ~10¹¹-fold. This observation suggests that the Zn²⁺ bimetallo core of AP superfamily enzymes, which is equipotent in phosphate monoester and diester catalysis, has the potential to become specialized for the hydrolysis of each class of phosphate esters via addition

  11. Site-directed mutagenesis maps interactions that enhance cognate and limit promiscuous catalysis by an alkaline phosphatase superfamily phosphodiesterase.

    PubMed

    Wiersma-Koch, Helen; Sunden, Fanny; Herschlag, Daniel

    2013-12-23

    Catalytic promiscuity, an evolutionary concept, also provides a powerful tool for gaining mechanistic insights into enzymatic reactions. Members of the alkaline phosphatase (AP) superfamily are highly amenable to such investigation, with several members having been shown to exhibit promiscuous activity for the cognate reactions of other superfamily members. Previous work has shown that nucleotide pyrophosphatase/phosphodiesterase (NPP) exhibits a >10⁶-fold preference for the hydrolysis of phosphate diesters over phosphate monoesters, and that the reaction specificity is reduced 10³-fold when the size of the substituent on the transferred phosphoryl group of phosphate diester substrates is reduced to a methyl group. Here we show additional specificity contributions from the binding pocket for this substituent (herein termed the R' substituent) that account for an additional ~250-fold differential specificity with the minimal methyl substituent. Removal of four hydrophobic side chains suggested on the basis of structural inspection to interact favorably with R' substituents decreases phosphate diester reactivity 10⁴-fold with an optimal diester substrate (R' = 5'-deoxythymidine) and 50-fold with a minimal diester substrate (R' = CH₃). These mutations also enhance the enzyme's promiscuous phosphate monoesterase activity by nearly an order of magnitude, an effect that is traced by mutation to the reduction of unfavorable interactions with the two residues closest to the nonbridging phosphoryl oxygen atoms. The quadruple R' pocket mutant exhibits the same activity toward phosphate diester and phosphate monoester substrates that have identical leaving groups, with substantial rate enhancements of ~10¹¹-fold. This observation suggests that the Zn²⁺ bimetallo core of AP superfamily enzymes, which is equipotent in phosphate monoester and diester catalysis, has the potential to become specialized for the hydrolysis of each class of phosphate esters via addition

  12. 21 CFR 520.823 - Erythromycin phosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    .... (a) Specifications. Erythromycin phosphate is the phosphate salt of the antibiotic substance produced by the growth of Streptomyces erythreus or the same antibiotic substance produced by any other...

  13. 21 CFR 520.823 - Erythromycin phosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    .... (a) Specifications. Erythromycin phosphate is the phosphate salt of the antibiotic substance produced by the growth of Streptomyces erythreus or the same antibiotic substance produced by any other...

  14. 21 CFR 520.823 - Erythromycin phosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    .... (a) Specifications. Erythromycin phosphate is the phosphate salt of the antibiotic substance produced by the growth of Streptomyces erythreus or the same antibiotic substance produced by any other...

  15. 21 CFR 520.823 - Erythromycin phosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    .... (a) Specifications. Erythromycin phosphate is the phosphate salt of the antibiotic substance produced by the growth of Streptomyces erythreus or the same antibiotic substance produced by any other...

  16. 21 CFR 520.823 - Erythromycin phosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    .... (a) Specifications. Erythromycin phosphate is the phosphate salt of the antibiotic substance produced by the growth of Streptomyces erythreus or the same antibiotic substance produced by any other...

  17. Quality control in production of suspensions from solid ammonium phosphates (monoammonium phosphate and diammonium phosphate). [Monoammonium phosphate; diammonium phosphate

    SciTech Connect

    Achorn, F.P.; Balay, H.L.

    1982-01-01

    Suspensions of good quality can be produced from MAP and DAP. Suspension quality depends on the amount of impurities in the ammonium phosphate solids used. Tests have shown that adding ammonium fluoride helps lower viscosity of suspensions containing a considerable amount of impurities. Also, adding polyphosphates (such as 10-34-0, 9-32-0, and 11-37-0) as a source of part of the P/sub 2/O/sub 5/ (6 to 15% polyphosphate in the product) helps to produce a suspension that has excellent storage characteristics. When the polyphosphate content of the product (11-33-0 suspension) is between 10 and 15% it usually will not solidify during cold weather storage. Freight and production costs of granular ammonium phosphates are relatively low compared to other sources of P/sub 2/O/sub 5/ for the fluid fertilizer market; therefore, using MAP and DAP to produce suspensions is expected to continue to grow in popularity. 2 refs., 7 figs., 1 tab.

  18. Downscaling Alkaline Phosphatase Activity in a Subtropical Reservoir

    NASA Astrophysics Data System (ADS)

    Tseng, Y.

    2011-12-01

    This research was conducted by downscaling study to understand phosphorus (P)-deficient status of different plankton and the role of alkaline phosphatase activity (APA) in subtropical Feitsui Reservoir. Results from field survey showed that bulk APA (1.6~95.2 nM h-1) was widely observed in the epilimnion (0~20 m) with an apparent seasonal variations, suggesting that plankton in the system were subjected to P-deficient seasonally. Mixed layer depth (an index of phosphate availability) is the major factor influencing the variation of bulk APA and specific APA (124~1,253 nmol mg C-1 h-1), based on multiple linear regression analysis. Size-fractionated APA assays showed that picoplankton (size 0.2~3 um) contributed most of the bulk APA in the system. In addition, single-cell APA detected by enzyme-labeled fluorescence (ELF) assay indicated that heterotrophic bacteria are the major contributors of APA. Thus, we can infer that bacteria play an important role in accelerating P-cycle within P-deficient systems. Light/nutrient manipulation bioassays showed that bacterial growth was directly controlled by phosphate, while picocyanobacterial growth is controlled by light and can out-compete bacteria under P-limited condition with the aid of light. Further analysis revealed that the strength of summer typhoon is a factor responsible for the inter-annual variability of bulk and specific APA. APA study demonstrated the episodic events (e.g. strong typhoon and extreme precipitation) had significant influence on APA variability in sub-tropical to tropical aquatic ecosystems. Hence, the results herein will allow future studies on monitoring typhoon disturbance (intensity and frequency) as well as the APA of plankton during summer-to-autumn in subtropical systems.

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

    PubMed

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

    2007-08-01

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

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

    PubMed

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

    2016-08-10

    Hydrogels composed of two-dimensional (2D) nanomaterials have become an important alternative to replace traditional inorganic scaffolds for tissue engineering. Here, we describe a novel nanocrystalline material with 2D morphology that was synthesized by tuning the crystallization of the sodium-magnesium-phosphate system. We discovered that the sodium ion can regulate the precipitation of magnesium phosphate by interacting with the crystal's surface causing a preferential crystal growth that results in 2D morphology. The 2D nanomaterial gave rise to a physical hydrogel that presented extreme thixotropy, injectability, biocompatibility, bioresorption, and long-term stability. The nanocrystalline material was characterized in vitro and in vivo and we discovered that it presented unique biological properties. Magnesium phosphate nanosheets accelerated bone healing and osseointegration by enhancing collagen formation, osteoblasts differentiation, and osteoclasts proliferation through up-regulation of COL1A1, RunX2, ALP, OCN, and OPN. In summary, the 2D magnesium phosphate nanosheets could bring a paradigm shift in the field of minimally invasive orthopedic and craniofacial interventions because it is the only material available that can be injected through high gauge needles into bone defects in order to accelerate bone healing and osseointegration. PMID:27280476

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

    PubMed

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

    2015-01-01

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

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

    PubMed Central

    Xiao, Yutao; Zhang, Dandan; Zhang, Yongdong; Li, Xianchun; Tabashnik, Bruce E.; Wu, Kongming

    2015-01-01

    Evolution of resistance by insects threatens the continued success of pest control using insecticidal crystal (Cry) proteins from the bacterium Bacillus thuringiensis (Bt) in sprays and transgenic plants. In this study, laboratory selection with Cry1Ac yielded five strains of cotton bollworm, Helicoverpa armigera, with resistance ratios at the median lethal concentration (LC50) of activated Cry1Ac ranging from 22 to 1700. Reduced activity and reduced transcription of an alkaline phosphatase protein that binds Cry1Ac was associated with resistance to Cry1Ac in the four most resistant strains. A Cry1Ac-binding fragment of alkaline phosphatase from H. armigera (HaALP1f) was not toxic by itself, but it increased mortality caused by Cry1Ac in a susceptible strain and in all five resistant strains. Although synergism of Bt toxins against susceptible insects by toxin-binding fragments of cadherin and aminopeptidase N has been reported previously, the results here provide the first evidence of synergism of a Bt toxin by a toxin-binding fragment of alkaline phosphatase. The results here also provide the first evidence of synergism of a Bt toxin by any toxin-binding peptide against resistant insects. PMID:25885820

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

    PubMed

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

    2015-01-01

    Evolution of resistance by insects threatens the continued success of pest control using insecticidal crystal (Cry) proteins from the bacterium Bacillus thuringiensis (Bt) in sprays and transgenic plants. In this study, laboratory selection with Cry1Ac yielded five strains of cotton bollworm, Helicoverpa armigera, with resistance ratios at the median lethal concentration (LC50) of activated Cry1Ac ranging from 22 to 1700. Reduced activity and reduced transcription of an alkaline phosphatase protein that binds Cry1Ac was associated with resistance to Cry1Ac in the four most resistant strains. A Cry1Ac-binding fragment of alkaline phosphatase from H. armigera (HaALP1f) was not toxic by itself, but it increased mortality caused by Cry1Ac in a susceptible strain and in all five resistant strains. Although synergism of Bt toxins against susceptible insects by toxin-binding fragments of cadherin and aminopeptidase N has been reported previously, the results here provide the first evidence of synergism of a Bt toxin by a toxin-binding fragment of alkaline phosphatase. The results here also provide the first evidence of synergism of a Bt toxin by any toxin-binding peptide against resistant insects. PMID:25885820

  4. Natural gas seeps in the French Alps: Sources and pathways

    NASA Astrophysics Data System (ADS)

    Kloppmann, Wolfram; Blessing, Michaela; Proust, Eric; Gal, Frédéric; Bentivegna, Gaetan; Henry, Benoit; Defossez, Pierrick; Catherine, Lerouge; Humez, Pauline; Mayer, Bernhard; Millot, Romain; Gaucher, Eric

    2016-04-01

    Natural gas emanations are part of the geochemical baseline to take into account when assessing global greenhouse gas emissions and potential impacts of conventional and unconventional gas exploration and exploitation on groundwater. Examples of such natural gas macro-seeps are known in several parts of the world (Etiope et al., 2009). Only a limited number of them have been characterized for their gas and isotopic compositions. Such analyses can provide essential information for baseline studies, providing insight in the sources (biogenic vs. thermogenic or modified thermogenic) and pathways of such seeps and may allow for distinction of natural seeps from stray gas leakage associated with human activities. Here, we report gas concentrations and multi-isotope data (δ13C and δ2H of methane and ethane, δ13C and δ18O of CO2, 3He/4He ratio) of two gas seeps in the French subalpine chains, both in a similar geological and structural position within Middle Jurassic claystones along the eastern border of the large synclinal structures of the Vercors and the Chartreuse massifs (Moss, 1992). The "ardent fountain" (fontaine ardente) of Le Gua, 30 km south of Grenoble has most likely the longest continuous written record of existence of any individual natural gas seep, mentioned explicitly as early as the first quarter of the 5th century (Augustin of Hippo (St. Augustin), approx. 426) This natural seep was described in the past as a "wet seep" associated with a spring, whereas the second investigated seep, Rochasson near Meylan north of Grenoble, is a dry seep. Both seeps contain methane and ethane with thermogenic C and H isotope signatures, comparable with a seep in the Northern Swiss Alps at Giswil (Etiope et al., 2010) but with a higher dryness (C1/(C2+C3)>1000) for the Le Gua seep, possibly due to molecular fractionation upon advective fluid+gas migration (Etiope et al., 2009). Maturity (R0) of the reservoir rocks deduced from δ13C(CH4), δ13C(C2H6) is similar to

  5. An inventory of glacial lakes in the Austrian Alps

    NASA Astrophysics Data System (ADS)

    Buckel, Johannes; Otto, Jan-Christoph; Keuschnig, Markus; Götz, Joachim

    2016-04-01

    The formation of lakes is one of the consequences of glacier retreat due to climate change in mountain areas. Numerous lakes have formed in the past few decades in many mountain regions around the globe. Some of these lakes came into focus due to catastrophic hazard events especially in the Himalayas and the Andes. Glacial lake development and lifetime is controlled by the complex interplay of glacier dynamics, geomorphological process activity and geological boundary conditions. Besides the hazard potential new lakes in formerly glaciated areas will significantly contribute to a new landscape setting and to changing geomorphologic, hydrologic and ecologic conditions at higher alpine altitudes. We present an inventory of high alpine lakes in the Austrian Alps located above an altitude of 1700 m asl. Most of these lakes are assumed to be of glacial origin, but other causes for development, like mass movements are considered as well. The inventory is a central part of the project FUTURELAKES that aims at modelling the potential development of glacial lakes in Austria (we refer to the presentation by Helfricht et al. during the conference for more details on the modelling part). Lake inventory data will serve as one basis for model validation since modelling is performed on different time steps using glacier inventory data. The purpose of the lake inventory is to get new insights into boundary conditions for lake formation and evolution by analysing existing lake settings. Based on these information the project seeks to establish a model of lake sedimentation after glacier retreat in order to assess the potential lifetime of the new lakes in Austria. Lakes with a minimum size of 1000 m² were mapped using multiple aerial imagery sources. The dataset contains information on location, geometry, dam type, and status of sedimentation for each lake. Additionally, various geologic, geomorphic and morphometric parameters describe the lake catchments. Lake data is related to

  6. Dating the polyphase evolution of the Western Alps

    NASA Astrophysics Data System (ADS)

    Villa, Igor M.

    2016-04-01

    A traverse through the Western Alps from the Zone Houillère to the Monte Rosa eclogites includes an oceanic basin (filled with Hercynian detrital muscovite) which was subducted during Alpine convergence and eclogitized; the eclogites were sheared in greenschist facies and exhumed to the surface. In the W the Zone Houillère sediments are weakly metamorphosed; further ESE, in the Ruitor Unit, higher strain rates and high pressure parageneses are observed; the Piemont-Ligurian (PL) Oceanic Unit consists of eclogites. Lu-Hf and Sm-Nd dating of prograde (55±4 Ma) and retrograde (41±7 Ma) garnet (Villa et al, JPetrol 55 (2014) 803) constrains eclogitization to the Eocene. 39Ar-40Ar ages of white mica show a spatial regularity: Zone Houillère (T ≈ 300 °C) muscovite ages are 270-100 Ma, Ar loss reaching 80 %; eclogitic phengite ages (P¬L Unit, T ≈ 500 °C, and Monte Rosa, T ≥ 600 °C) are constant at 47 Ma, independently of peak T, and agree with garnet Lu-Hf ages; Ruitor white mica ages depend on total strain and especially on the degree of phengitic substitution in detrital muscovite. The observation that Ar can be lost at T < 300 °C while at the same time it can be retained at T > 500-600 °C appears counterintuitive if one assumes that ages are a unique function of T. The Zone Houillère pattern is explained by sedimentology: during diagenesis, sediments are dehydrated, and large amounts of circulating water very efficiently promote recrystallization and Ar loss. In the Ruitor Unit, an different eastward age decrease is observed: mica rejuvenation correlates with phengite content, whereby HP phengite replaced detrital muscovite. These samples record a progressive increase of retrogression reactions that produce a heterochemical replacement mineral with associated Ar loss. The Cl/K ratio in white mica is an essential discriminator, as retrogression can be associated with more or less saline fluids. In the Monte Rosa and P-L Oceanic Units, the constant 47

  7. Eclogitic metatrondhjemites from metaophiolites of the Western Alps

    NASA Astrophysics Data System (ADS)

    Martin, Silvana; Tartarotti, Paola; Meyzen, Chrstine; Benciolini, Luca; Toffolo, Luca

    2016-04-01

    Eclogitic metatrondhjemites from metaophiolites of the Western Alps Martin S.**, Tartarotti P.*, Meyzen C. **, Benciolini L.***, Toffolo L. ** *Dipartimento di Scienze della Terra, Università degli Studi di Milano ** Dipartimento di Geoscienze, Università di Padova *** Dipartimento di Chimica, Fisica e Ambiente, Università di Udine In the Urtier valley (southern Aosta Valley, Italy), the Piemonte metaophiolites mainly consist of serpentinized peridotites including pods and boudinaged layers of Fe-metagabbro and trondhjemite transposed in the main eclogitic foliation. The contact between serpentinized peridotites and Fe-metagabbro/trondhjemite is locally lined by chloriteschist and rodingite. The high pressure parageneses in the Fe-metagabbro are omphacite-garnet-rutile-glaucophane-phengite, and in the metatrondhjemite plagioclase-quartz-phengite-clinozoisite-epidote-garnet, respectively. Bulk-rock major and trace elements in addition to O isotope analyses were performed in both rock types. Fe-metagabbros are characterized by MgO wt% ranging between 6.11 and 9.63%, ∑REE= 20-101 ppm, (La/Yb)N = 0.22-0.91; trondhjemites have SiO2 43%, Al2O3 ranging between 21 and 24%, CaO ranging between 17 and 20%, ∑REE = 172 - 272 ppm, (La/Yb)N ranging between 7.78 and 13.70. The δ18O is 5.9 ‰ in a Fe-metagabbro sample and 7.4 ‰ in a trondhjemite sample, suggesting that these rocks have been affected by a weak oceanic low temperature alteration. The high CaO content may indicate a metasomatic process which could have occurred during the oceanic stage or at high pressure conditions.

  8. Infrasonic monitoring of snow avalanches in the Alps

    NASA Astrophysics Data System (ADS)

    Marchetti, E.; Ulivieri, G.; Ripepe, M.; Chiambretti, I.; Segor, V.

    2012-04-01

    Risk assessment of snow avalanches is mostly related to weather conditions and snow cover. However a robust risk validation requires to identify all avalanches occurring, in order to compare predictions to real effects. For this purpose on December 2010 we installed a permanent 4-element, small aperture (100 m), infrasound array in the Alps, after a pilot experiment carried out in Gressonay during the 2009-2010 winter season. The array has been deployed in the Ayas Valley, at an elevation of 2000 m a.s.l., where natural avalanches are expected and controlled events are regularly performed. The array consists into 4 Optimic 2180 infrasonic microphones, with a sensitivity of 10-3 Pa in the 0.5-50 Hz frequency band and a 4 channel Guralp CMG-DM24 A/D converter, sampling at 100 Hz. Timing is achieved with a GPS receiver. Data are transmitted to the Department of Earth Sciences of the University of Firenze, where data is recorded and processed in real-time. A multi-channel semblance is carried out on the continuous data set as a function of slowness, back-azimuth and frequency of recorded infrasound in order to detect all avalanches occurring from the back-ground signal, strongly affected by microbarom and mountain induced gravity waves. This permanent installation in Italy will allow to verify the efficiency of the system in short-to-medium range (2-8 km) avalanche detection, and might represent an important validation to model avalanches activity during this winter season. Moreover, the real-time processing of infrasonic array data, might strongly contribute to avalanche risk assessments providing an up-to-description of ongoing events.

  9. Preparation and characterization of a degradable magnesium phosphate bone cement

    PubMed Central

    Yu, Ying; Xu, Chao; Dai, Honglian

    2016-01-01

    A kind of degradable magnesium phosphate bone cement (MPBC) was fabricated by using the mixed powders of magnesium oxide (MgO), potassium dihydrogen phosphate (KH2PO4) and calcium dihydrogen phosphate (Ca(H2PO4)2.H2O). As MgKPO4, the main product of MgO and KH2PO4 was alkaline, the Ca(H2PO4)2.H2O was added to neutralize the alkali of the system. And the effects of Ca(H2PO4)2.H2O on the performance of MPBC were discussed. The results showed that the adding of Ca(H2PO4)2.H2O extended the setting time, which was about 6 min to 18 min. The compressive strength increased first and then decreased, and maximum value reached 31.2 MPa after setting for 24 h without any additional pressure. The MPBC was degradable in Tris–HCl solution, and the extracts of the cytotoxicity assay showed that the MPBC had good biocompatibility, indicating that the MPBC had good biodegradable and biocompatible properties. PMID:27482465

  10. Preparation and characterization of a degradable magnesium phosphate bone cement.

    PubMed

    Yu, Ying; Xu, Chao; Dai, Honglian

    2016-12-01

    A kind of degradable magnesium phosphate bone cement (MPBC) was fabricated by using the mixed powders of magnesium oxide (MgO), potassium dihydrogen phosphate (KH2PO4) and calcium dihydrogen phosphate (Ca(H2PO4)2.H2O). As MgKPO4, the main product of MgO and KH2PO4 was alkaline, the Ca(H2PO4)2.H2O was added to neutralize the alkali of the system. And the effects of Ca(H2PO4)2.H2O on the performance of MPBC were discussed. The results showed that the adding of Ca(H2PO4)2.H2O extended the setting time, which was about 6 min to 18 min. The compressive strength increased first and then decreased, and maximum value reached 31.2 MPa after setting for 24 h without any additional pressure. The MPBC was degradable in Tris-HCl solution, and the extracts of the cytotoxicity assay showed that the MPBC had good biocompatibility, indicating that the MPBC had good biodegradable and biocompatible properties. PMID:27482465

  11. Antimicrobial potency of alkali ion substituted calcium phosphate cements.

    PubMed

    Gbureck, Uwe; Knappe, Oliver; Grover, Liam M; Barralet, Jake E

    2005-12-01

    Potassium and sodium containing nanoapatite cements were produced by the reaction of mechanically activated CaNaPO(4) (CSP), CaKPO(4) (CPP) and Ca(2)KNa(PO(4))(2) (CPCP) with a 2.5% Na(2)HPO(4) solution. The cements exhibited clinically acceptable setting times of approximately 5 min and compressive strengths of 5-10 MPa. The antimicrobial properties of the cements were tested with the agar diffusion test using Streptococcus salvarius, Staphylococcus epidermis and Candida albicans. All types of alkali ion containing cements showed a significantly higher antimicrobial potency with inhibition zones of approx. 4-11 mm than a commercial calcium hydroxide cement which resulted in small inhibition zones around the cement samples of a maximum of 1.5 mm. The antimicrobial properties of all the cements were not found to diminish even after longer incubation times. This behaviour was attributed to the formation of soluble alkaline metal phosphates during setting which increased the pH value in the agar gel around the alkali containing calcium phosphate cement to 8.5-10.7 compared to 6.5-8.0 for the Ca(OH)(2) product. The high antimicrobial potency of alkali-calcium phosphate cements may find an application in dentistry as pulp capping agents, root fillers or cavity liners. PMID:16005511

  12. "Wrong-way" subduction of South Alpine (Adriatic) lithosphere beneath the Eastern Alps - a kinematic appraisal

    NASA Astrophysics Data System (ADS)

    Handy, Mark R.; Ustaszewski, Kamil; Kissling, Eduard; Schmid, Stefan M.; Rosenberg, Claudio L.

    2013-04-01

    The junction of the Central and Eastern Alps coincides with a switch in subduction polarity as indicated by seismic tomography with high resolution locally down to 400 km (Lippitsch et al. 2003). Whereas the SE dip of the positive Vp slab anomaly beneath the Central Alps is consistent with subduction of European lithosphere, the correspondingly steep N to NE dip of the anomaly beneath the Eastern Alps has lead to speculation that it represents either Adriatic lithosphere subducted the "wrong way" beneath the Alpine orogen during Miocene oblique dextral convergence in the Dinarides (Kissling et al. 2006, Ustaszewski et al. 2008) or European lithosphere that was steepened and overturned during Miocene shortening of the Eastern Alps (Mitterbauer et al. 2011). We tested the kinematic plausibility of an Adriatic origin for the slab beneath the Eastern Alps by straightening and horizontalizing its leading edge, then moving this edge backward to locations in Cenozoic to Late Cretaceous time. The motion is constrained by crustal shortening and extensional values across the Alpine chain which are applied as successive retrotranslations and backrotations of points on stable parts of the Adriatic microplate with respect to a European reference frame. This procedure yields a series of paleotectonic maps for critical time slices (20, 35, 67, 84 Ma) that show the location of the slab edge with respect to the evolving Alps-Carpathian-Dinaric Orogen. Our reconstruction reveals that from 84 to 20 Ma the horizontalized northern edge of the slab was located beneath the Eastern Alps, just N of the current trace of the Periadriatic Fault. The eastern edge of the slab is interpreted to have delimited a major dextral transfer fault (Alps-Dinarides Transfer, ADT) that linked 84-35 Ma SE-directed subduction of Alpine Tethys and the European margin with oblique, N- to NE-directed subduction of Neotethys and the Adriatic margin. Post-35 Ma collision in the Alps triggered breakoff of the

  13. Formation of the arc of the Western Alps and Alps-Apennines transition in the light of new geophysical data on the lithospheric architecture around the Ligurian knot

    NASA Astrophysics Data System (ADS)

    Kissling, E.; Bousquet, R.; Ford, M.; Schmid, S.

    2012-04-01

    This study integrates new geophysical data on the lithospheric structure of the Western Alps with a wealth of geologic and tectonic data. Geological evidence suggests an evolution of the Western Alps in 3 stages: Stage 1 (85-35 Ma) is dominated by NW- to NNW-directed convergence of Adria with respect to Europe, leading to subduction in the Central Alps and sinistral transpression in the Western Alps; collisional stage 2 (35-23Ma) results in top WNW out of sequence thrusting along the Penninic front associated with some 240km of WNW-directed indentation of the Ivrea zone at the tip of the Adria plate into the Western Alpine orogen, substantially modifying its geometry in terms of post-nappe folding; stage 3 (23 Ma to recent) involves a change in subduction polarity in the Ligurian Alps and Apennines, linked to the modification of the southern part of the Western Alps in terms of oroclinal bending during rotational roll-back of the Adria plate around a pole of rotation ("Ligurian knot"). Initiated in Oligocene times this rotation created the the young Ligurian plate . New geophysical data (controlled source seismology, local earthquake tomography and receiver functions) tightly constrain the geometry of the Moho around the triple junction area between Europe, Adria and the new Ligurian plate that came into existence at the expense of the European plate near its suture with Adria. Moreover, these new data constrain the 3D geometry of the geophysical Ivrea body associated with a wedge of subcontinental mantle belonging to the Adria plate that rose to the surface of the earth in parts of the Western Alps during Alpine convergence. The internal (eastern) top of the Ivrea mantle wedge is identical with the Adriatic Moho rising up to shallow crustal depth. Local earthquake tomography constrains the external (western) limit of the Ivrea mantle wedge for the first time. The Ivrea wedge's subvertical-to-steeply west-dipping interface with the Penninic nappes and the Alpine

  14. The effect of synthetic α-tricalcium phosphate on osteogenic differentiation of rat bone mesenchymal stem cells

    PubMed Central

    Liu, Jinzhong; Zhao, Liang; Ni, Ling; Qiao, Chunyan; Li, Daowei; Sun, Hongchen; Zhang, Zongtao

    2015-01-01

    The reconstruction of large bone defects has been the focus in bone tissue engineering research. By acting as synthetic frameworks for cell growth and tissue formation, biomaterials can play a critical role in bone tissue engineering. Among various biomaterials, calcium phosphate based materials include hydroxyapatite (HA), α-tricalcium phosphate (α-TCP), and β-tricalcium phosphate (β-TCP) are widely used as scaffold materials in bone tissue engineering. However, little is known about the effect of α-TCP alone on the osteogenic differentiation of the BMSCs. To this end, we synthesized α-TCP using a novel co-precipitation method. The synthetic α-TCP was then incubated with rat BMSCs under osteogenic inductive medium culture conditions, followed by the analysis of the mRNA levels of various osteogenesis-related genes, including ALP, Rux2, COL-I, and SP7, using a quantitative RT-PCR method. Following incubation of BMSCs with 20 μg/ml α-TCP, cells reached confluency after 7 days. Additionally, the MTT analysis showed that α-TCP at concentration of 10-20 μg/ml had good biocompatibility with BMSCs, showing no significant inhibition of rat BMSCs proliferation. Furthermore, the synthetic α-TCP (20 μg/ml), when incubated with rat BMSCs in the osteogenic culture medium, increased the mRNA levels of various osteogenesis-related genes, including ALP, Rux2, COL-I, and SP7. Finally, treatment of synthetic α-TCP (20 μg/ml) potentiated calcium nodule formations after incubation with rat BMSCs in osteogenic culture medium for 21 days, as compared with non-treated control. Taken together, the results in the present study suggested that α-TCP alone likely promotes rat BMSCs osteogenic differentiation through up-regulating ALP, Col-I, Runx2, and SP7 gene expression. PMID:26550458

  15. The effect of synthetic α-tricalcium phosphate on osteogenic differentiation of rat bone mesenchymal stem cells.

    PubMed

    Liu, Jinzhong; Zhao, Liang; Ni, Ling; Qiao, Chunyan; Li, Daowei; Sun, Hongchen; Zhang, Zongtao

    2015-01-01

    The reconstruction of large bone defects has been the focus in bone tissue engineering research. By acting as synthetic frameworks for cell growth and tissue formation, biomaterials can play a critical role in bone tissue engineering. Among various biomaterials, calcium phosphate based materials include hydroxyapatite (HA), α-tricalcium phosphate (α-TCP), and β-tricalcium phosphate (β-TCP) are widely used as scaffold materials in bone tissue engineering. However, little is known about the effect of α-TCP alone on the osteogenic differentiation of the BMSCs. To this end, we synthesized α-TCP using a novel co-precipitation method. The synthetic α-TCP was then incubated with rat BMSCs under osteogenic inductive medium culture conditions, followed by the analysis of the mRNA levels of various osteogenesis-related genes, including ALP, Rux2, COL-I, and SP7, using a quantitative RT-PCR method. Following incubation of BMSCs with 20 μg/ml α-TCP, cells reached confluency after 7 days. Additionally, the MTT analysis showed that α-TCP at concentration of 10-20 μg/ml had good biocompatibility with BMSCs, showing no significant inhibition of rat BMSCs proliferation. Furthermore, the synthetic α-TCP (20 μg/ml), when incubated with rat BMSCs in the osteogenic culture medium, increased the mRNA levels of various osteogenesis-related genes, including ALP, Rux2, COL-I, and SP7. Finally, treatment of synthetic α-TCP (20 μg/ml) potentiated calcium nodule formations after incubation with rat BMSCs in osteogenic culture medium for 21 days, as compared with non-treated control. Taken together, the results in the present study suggested that α-TCP alone likely promotes rat BMSCs osteogenic differentiation through up-regulating ALP, Col-I, Runx2, and SP7 gene expression. PMID:26550458

  16. Uptake of arsenic by alkaline soils near alkaline coal fly ash disposal facilities.

    PubMed

    Khodadoust, Amid P; Theis, Thomas L; Murarka, Ishwar P; Naithani, Pratibha; Babaeivelni, Kamel

    2013-12-01

    The attenuation of arsenic in groundwater near alkaline coal fly ash disposal facilities was evaluated by determining the uptake of arsenic from ash leachates by surrounding alkaline soils. Ten different alkaline soils near a retired coal fly ash impoundment were used in this study with pH ranging from 7.6 to 9.0, while representative coal fly ash samples from two different locations in the coal fly ash impoundment were used to produce two alkaline ash leachates with pH 7.4 and 8.2. The arsenic found in the ash leachates was present as arsenate [As(V)]. Adsorption isotherm experiments were carried out to determine the adsorption parameters required for predicting the uptake of arsenic from the ash leachates. For all soils and leachates, the adsorption of arsenic followed the Langmuir and Freundlich equations, indicative of the favorable adsorption of arsenic from leachates onto all soils. The uptake of arsenic was evaluated as a function of ash leachate characteristics and the soil components. The uptake of arsenic from alkaline ash leachates, which occurred mainly as calcium hydrogen arsenate, increased with increasing clay fraction of soil and with increasing soil organic matter of the alkaline soils. Appreciable uptake of arsenic from alkaline ash leachates with different pH and arsenic concentration was observed for the alkaline soils, thus attenuating the contamination of groundwater downstream of the retired coal fly ash impoundment.

  17. Reaching and abandoning the furthest ice extent during the Last Glacial Maximum in the Alps

    NASA Astrophysics Data System (ADS)

    Ivy-Ochs, Susan; Wirsig, Christian; Zasadni, Jerzy; Hippe, Kristina; Christl, Marcus; Akçar, Naki; Schluechter, Christian

    2016-04-01

    During the Last Glacial Maximum (LGM) in the European Alps (late Würm) local ice caps and extensive ice fields in the high Alps fed huge outlet glaciers that occupied the main valleys and extended onto the forelands as piedmont lobes. Records from numerous sites suggest advance of glaciers beyond the mountain front by around 30 ka (Ivy-Ochs 2015 and references therein). Reaching of the maximum extent occurred by about 27-26 ka, as exemplified by dates from the Rhein glacier area (Keller and Krayss, 2005). Abandonment of the outermost moraines at sites north and south of the Alps was underway by about 24 ka. In the high Alps, systems of transection glaciers with transfluences over many of the Alpine passes dominated, for example, at Grimsel Pass in the Central Alps (Switzerland). 10Be exposure ages of 23 ± 1 ka for glacially sculpted bedrock located just a few meters below the LGM trimline in the Haslital near Grimsel Pass suggest a pulse of ice surface lowering at about the same time that the foreland moraines were being abandoned (Wirsig et al., 2016). Widespread ice surface lowering in the high Alps was underway by no later than 18 ka. Thereafter, glaciers oscillated at stillstand and minor re-advance positions on the northern forelands for several thousand years forming the LGM stadial moraines. Final recession back within the mountain front took place by 19-18 ka. Recalculation to a common basis of all published 10Be exposure dates for boulders situated on LGM moraines suggests a strong degree of synchrony for the timing of onset of ice decay both north and south of the Alps. Ivy-Ochs, S., 2015, Cuadernos de investigación geográfica 41: 295-315. Keller, O., Krayss, E., 2005, Vierteljahrschr. Naturforsch. Gesell. Zürich 150: 69-85. Wirsig, C. et al., 2016, J. Quat. Sci. 31: 46-59.

  18. A simple-potentiometric method for determination of acid and alkaline phosphatase enzymes in biological fluids and dairy products using a nitrophenylphosphate plastic membrane sensor.

    PubMed

    Hassan, Saad S M; Sayour, Hossam E M; Kamel, Ayman H

    2009-04-27

    A novel poly(vinyl chloride) matrix membrane sensor responsive to 4-nitrophenylphosphate (4-NPP) substrate is described, characterized and used for the potentiometric assay of acid (ACP) and alkaline (ALP) phosphatase enzymes. The sensor is based on the use of the ion-association complex of 4-NPP anion with nickel(II)-bathophenanthroline cation as an electroactive material and nitrophenyloctyl ether (NPOE) as a solvent mediator. The sensor displays good selectivity and stability and demonstrates a near-Nernstian response for 4-NPP over the concentration range 9.6x10(-6) to 1.0x10(-2) M with an anionic slope of 28.6+/-0.3 mV decade(-1) and a detection limit of 6.3x10(-6) M over the pH range 4.5-10. The sensor is used to measure the decrease of a fixed concentration of 4-NPP substrate as a function of acid and alkaline phosphatase enzyme activities at optimized conditions of pH and temperature. A linear relationship between the initial rate of 4-NPP substrate hydrolysis and enzyme activity holds over 0.05-3.0 and 0.03-3.4 IU L(-1) of ACP and ALP enzymes, respectively. Validation of the method by measuring the lower detection limit, range, accuracy, precision, within-day repeatability and between-day-variability reveals good performance characteristics of the proposed sensor. The sensor is used for the determination of acid and alkaline phosphatase enzyme activities in biological fluids of some patients suffering from alcoholic cirrhosis, acute myelocytic leukemia, pre-eclampsia and prostatic cancer. The sensor is also utilized for assessment of alkaline phosphatase enzyme in milk and dairy products. The results obtained agree fairly well with data obtained by the standard spectrophotometric methods.

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

    PubMed

    Majumder, Shyam Prasad; Das, Amal Chandra

    2016-04-01

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

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

    PubMed

    Majumder, Shyam Prasad; Das, Amal Chandra

    2016-04-01

    An experiment was conducted under laboratory conditions to investigate the effect of four organophosphate insecticides, viz. monocrotophos, profenophos, quinalphos and triazophos at their field application rates (0.75, 1.0, 0.5 and 0.6 kg a.i.ha(-1), respectively), on the growth and activities of phosphate solubilizing microorganisms in relation to availability of insoluble phosphates in the Gangetic alluvial soil of West Bengal, India. The proliferation of phosphate solubilizing microorganisms was highly induced with profenophos (38.3%), while monocrotophos exerted maximum stimulation (20.8%) towards the solubility of insoluble phosphates in soil. The phosphatase activities of the soil (both acid phosphatase and alkaline phosphatase) were significantly increased due to the incorporation of the insecticides in general, and the augmentation was more pronounced with quinalphos (43.1%) followed by profenophos (27.6%) for acid phosphatase, and with monocrotophos (25.2%) followed by profenophos (16.1%) for alkaline phosphatase activity in soil. The total phosphorus was highly retained by triazophos (19.9%) followed by monocrotophos (16.5%), while incorporation of triazophos and quinalphos manifested greater availability of water soluble phosphorus in soil. PMID:26720809

  1. Effect of different calcium phosphate scaffold ratios on odontogenic differentiation of human dental pulp cells.

    PubMed

    AbdulQader, Sarah Talib; Kannan, Thirumulu Ponnuraj; Rahman, Ismail Ab; Ismail, Hanafi; Mahmood, Zuliani

    2015-04-01

    Calcium phosphate (CaP) scaffolds have been widely and successfully used with osteoblast cells for bone tissue regeneration. However, it is necessary to investigate the effects of these scaffolds on odontoblast cells' proliferation and differentiation for dentin tissue regeneration. In this study, three different hydroxyapatite (HA) to beta tricalcium phosphate (β-TCP) ratios of biphasic calcium phosphate (BCP) scaffolds, BCP20, BCP50, and BCP80, with a mean pore size of 300μm and 65% porosity were prepared from phosphoric acid (H2PO4) and calcium carbonate (CaCO3) sintered at 1000°C for 2h. The extracts of these scaffolds were assessed with regard to cell viability and differentiation of odontoblasts. The high alkalinity, more calcium, and phosphate ions released that were exhibited by BCP20 decreased the viability of human dental pulp cells (HDPCs) as compared to BCP50 and BCP80. However, the cells cultured with BCP20 extract expressed high alkaline phosphatase activity and high expression level of bone sialoprotein (BSP), dental matrix protein-1 (DMP-1), and dentin sialophosphoprotein (DSPP) genes as compared to that cultured with BCP50 and BCP80 extracts. The results highlighted the effect of different scaffold ratios on the cell microenvironment and demonstrated that BCP20 scaffold can support HDPC differentiation for dentin tissue regeneration.

  2. A multidisciplinary methodology for the characterization of a large rock spread in the Northern Calcareous Alps (Eastern Alps)

    NASA Astrophysics Data System (ADS)

    Melzner, Sandra; Ottowitz, David; Pfeiler, Stefan; Moser, Michael; Motschka, Klaus; Lotter, Michael; Mandl, Gerhard Walter; Rohn, Joachim; Otter, Juergen; Wimmer-Frey, Ingeborg

    2015-04-01

    The Northern Calcareous Alps are characterized by complex lithological and tectonic settings, which are a consequence of the multiphase Alpine orogeny. Several tectonic events caused a varying structural anisotropy with a high susceptibility towards certain types of gravitational mass movement. Mt. Plassen is situated west of the Hallstatt village (Upper Austria). It is composed of Jurassic limestone, which overlies Permotriassic fine-grained clastic rocks and evaporites (mainly part of the so-called Haselgebirge). This geotechnical predisposition causes rock spreading of the more hard and rigid limestone on the weak, mainly clayey rocks. Associated to this large slope instability are secondary rockfall and sliding processes. Further common process chains include rockfall triggering slides and/or earth flows by undrained loading of the ductile clay material. Thus, such fast moving flows/slides may endanger the houses and infrastructures in the Salzberg high valley and Hallstatt village. Recent rockfall activity at Mt. Plassen provide evidences for greater, perhaps accelerating displacement rates of the rock spread. A multidisciplinary assessment strategy was chosen to analyse the ground conditions, to characterize the potential failure mechanisms in more detail and to evaluate the hazard potential of future events. Methods include field mapping (geologic, engineering geologic and geomorphologic), sampling and determination of soil parameters in active process areas, geophysical surveys (airborne geophysics and geoelectric measurements) and kinematic measurements (tape dilatometer and geodetic measurements over a period of 50 years). Results of this multidisciplinary approach form the basis for further decision making such as the installation of a monitoring system or other preventive measures.

  3. [Increase in Alkaline Phosphatase Activity after High-Fat Meal Ingestion is Correlated to the Amount of ABH Substances in Saliva].

    PubMed

    Matsushita, Makoto; Otani, Kana; Sakamoto, Yui; Arai, Tomoko; Yukimasa, Nobuyasu; Muramoto, Yoshimi; Komoda, Tsugikazu

    2015-05-01

    Intestinal alkaline phosphatase (IAP) appears in the circulation more frequently in blood group B or O secretors than in blood group A or AB secretors and non-secretors, and serum IAP activity rises following the ingestion of a high-fat meal. In a previous study, the occurrence of two IAP isoforms, with high (HIAP) and normal molecular mass (NIAP), in healthy sera was demonstrated by 6.0% polyacrylamide gel electrophoresis in the presence of 1% Triton X-100. NIAP was present in the fasting serum of only healthy blood group B or O secretors, but was present in all subjects following ingestion of a high-fat meal. We classified 56 healthy subjects into 3 blood groups: B (n = 19), O (n = 17), and A (n = 20), and measured their serum ALP activity in a fasting state and 6 h after a high-fat meal. The amount of ABH substances in the saliva of each subject was determined by the hemagglutination inhibition test. Correlation coefficients between the change in ALP activity after high-fat meal ingestion and the hemagglutination inhibition values in saliva were 0.925 in blood group B, 0.879 in blood group O, and 0.906 in blood group A. These results suggest that increases in ALP activity in the circulation following the ingestion of a high-fat meal are closely related to the amount of ABH substances in saliva. PMID:26524892

  4. Toxicological review of inorganic phosphates.

    PubMed

    Weiner, M L; Salminen, W F; Larson, P R; Barter, R A; Kranetz, J L; Simon, G S

    2001-08-01

    Inorganic phosphate salts are widely used as food ingredients and in a variety of commercial applications. The United States Food and Drug Administration (FDA) considers inorganic phosphates "Generally Recognized As Safe" (GRAS) (FDA, 1973a, 1979) [FDA: Food and Drug Administration 1973a. GRAS (Generally Recognized as Safe) food ingredients-phosphates. NTIS PB-221-224, FDA, Food and Drug Administration, 1979. Phosphates; Proposed Affirmation of and Deletion From GRAS Status as Direct and Human Food Ingredients. Federal Register 44 (244). 74845-74857, 18 December (1979)] and the European Union (EU) allows inorganic phosphates to be added directly to food (EU Directive 95/2/EC as amended by 98/72/EC). In this review, data on the acute, subchronic and chronic toxicity, genotoxicity, teratogenicity and reproductive toxicity from the published literature and from unpublished studies by the manufacturers are reviewed. Based on the toxicity data and similar chemistry, the inorganic phosphates can be separated into four major classes, consisting of monovalent salts, divalent salts, ammonium salts and aluminum salts. The proposed classification scheme supports the use of toxicity data from one compound to assess the toxicity of another compound in the same class. However, in the case of eye and skin irritation, the proposed classification scheme cannot be used because a wide range of responses exists within each class. Therefore, the eye and skin hazards associated with an individual inorganic phosphate should be assessed on a chemical-by-chemical basis. A large amount of toxicity data exists for all four classes of inorganic phosphates. The large and comprehensive database allows an accurate assessment of the toxicity of each class of inorganic phosphate. Overall, all four classes of inorganic phosphates exhibit low oral, inhalation and dermal toxicities. Based on these data, humans are unlikely to experience adverse effects when the daily phosphorus consumption remains

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

    SciTech Connect

    Cambier, Linda; Pomies, Pascal

    2011-06-17

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

  6. Novel highly biodegradable biphasic tricalcium phosphates composed of alpha-tricalcium phosphate and beta-tricalcium phosphate.

    PubMed

    Li, Yanbao; Weng, Wenjian; Tam, Kim Chiu

    2007-03-01

    Novel biodegradable biphasic tricalcium phosphates (BTCP) composed of alpha-tricalcium phosphate (alpha-TCP) and beta-tricalcium phosphate (beta-TCP) were successfully synthesized by heating amorphous calcium phosphate precursors with different structures at 800 degrees C for 3 h. The ratio of alpha-TCP and beta-TCP in the calcium phosphate particle can be controlled by aging time and pH value during synthesis of the amorphous precursor.

  7. An efficient, multiply promiscuous hydrolase in the alkaline phosphatase superfamily

    PubMed Central

    van Loo, Bert; Jonas, Stefanie; Babtie, Ann C.; Benjdia, Alhosna; Berteau, Olivier; Hyvönen, Marko; Hollfelder, Florian

    2010-01-01

    We report a catalytically promiscuous enzyme able to efficiently promote the hydrolysis of six different substrate classes. Originally assigned as a phosphonate monoester hydrolase (PMH) this enzyme exhibits substantial second-order rate accelerations ((kcat/KM)/kw), ranging from 107 to as high as 1019, for the hydrolyses of phosphate mono-, di-, and triesters, phosphonate monoesters, sulfate monoesters, and sulfonate monoesters. This substrate collection encompasses a range of substrate charges between 0 and -2, transition states of a different nature, and involves attack at two different reaction centers (P and S). Intrinsic reactivities (half-lives) range from 200 days to 105 years under near neutrality. The substantial rate accelerations for a set of relatively difficult reactions suggest that efficient catalysis is not necessarily limited to efficient stabilization of just one transition state. The crystal structure of PMH identifies it as a member of the alkaline phosphatase superfamily. PMH encompasses four of the native activities previously observed in this superfamily and extends its repertoire by two further activities, one of which, sulfonate monoesterase, has not been observed previously for a natural enzyme. PMH is thus one of the most promiscuous hydrolases described to date. The functional links between superfamily activities can be presumed to have played a role in functional evolution by gene duplication. PMID:20133613