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Sample records for acid site strength

  1. Number and strength of surface acidic sites on porous aluminosilicates of the MCM-41 type inferred from a combined microcalorimetric and adsorption study

    SciTech Connect

    Meziani, M.J.; Zajac, J.; Jones, D.J.; Partyka, S.; Roziere, J.

    2000-03-07

    A combined microcalorimetry and adsorption study has been used to characterize the surface acidity of two series of MCM-41 aluminosilicates (referred to as SiAlxCn, where x is the mole Si:Al ration and n the chain length of the surfactant template). {sup 29}Si magic angle spinning NMR spectra of a selected sample (SiAl32C16) indicates the presence of siloxane groups, Si(OSi){sub 4}, and three types of silanol groups, that is, single (SiO){sub 3}-Si-OH, hydrogen-bonded (SiO){sub 3}-SiOH{hor{underscore}ellipsis}HO-Si-(SiO){sub 3}, and germinal (SiO){sub 2}-Si(OH){sub 2}. It is also possible to detect the contributions from Si(3Si, 1Al) and Si(2Si,2Al) sites. The volumetric and calorimetric measurements of gas ammonia adsorption at 353 K were used to determine the number and strength of surface acidic sites. With the exception of H{sup +}-SiAl32C14 and SiAl8C14, all samples have low surface acidity. Following the pyridine-TPD study on SiAl9C14 sample, Lewis acid sites producing surface pyridine complexes constitute the strongest acidic site.

  2. College Chemistry Students' Mental Models of Acids and Acid Strength

    ERIC Educational Resources Information Center

    McClary, LaKeisha; Talanquer, Vicente

    2011-01-01

    The central goal of this study was to characterize the mental models of acids and acid strength expressed by advanced college chemistry students when engaged in prediction, explanation, and justification tasks that asked them to rank chemical compounds based on their relative acid strength. For that purpose we completed a qualitative research…

  3. Acid Strength and Bifunctional Catalytic Behavior of Alloys Comprised of Noble Metals and Oxophilic Metal Promoters

    SciTech Connect

    Hibbitts, David D.; Tan, Qiaohua; Neurock, Matthew

    2014-06-01

    The promotion of metal catalysts with partially oxidized oxophilic MOx species, such as ReOx-promoted Rh, has been demonstrated to produce Brønsted acid sites that can promote hydrogenolysis of oxygenate intermediates such as those found in biomass-derived species. A wide variety of alloy compositions and structures are examined in this work to investigate strongly acidic promoters by using DFT-calculated deprotonation energies (DPE) as a measure of acid strength. Sites with the highest acid strength had DPE less than 1100 kJ mol-1, similar to DPE values of heteropolyacids or acid-containing zeolites, and were found on alloys composed of an oxophilic metal (such as Re or W) with a noble metal (such as Rh or Pt). NH3 adsorbs more strongly to sites with increasing acid strength and the activation barriers for acid-catalyzed ring opening of a furan ring decrease with increasing acid strength, which was also shown to be stronger for OH acid sites bound to multiple oxophilic metal atoms in a three-fold configuration rather than OH sites adsorbed in an atop configuration on one oxophilic metal, indicating that small MOx clusters may yield sites with the highest acid strength.

  4. Solvation and Acid Strength Effects on Catalysis by Faujasite Zeolites

    SciTech Connect

    Gounder, Rajamani P.; Jones, Andrew J.; Carr, Robert T.; Iglesia, Enrique

    2012-02-01

    Kinetic, spectroscopic, and chemical titration data indicate that differences in monomolecular isobutane cracking and dehydrogenation and methanol dehydration turnover rates (per H+) among FAU zeolites treated thermally with steam (H-USY) and then chemically with ammonium hexafluorosilicate (CDHUSY) predominantly reflect differences in the size and solvating properties of their supercage voids rather than differences in acid strength. The number of protons on a given sample was measured consistently by titrations with Na+, with CH3 groups via reactions of dimethyl ether, and with 2,6-di-tert-butylpyridine during methanol dehydration catalysis; these titration values were also supported by commensurate changes in acidic OH infrared band areas upon exposure to titrant molecules. The number of protons, taken as the average of the three titration methods, was significantly smaller than the number of framework Al atoms (Alf) obtained from X-ray diffraction and 27Al magic angle spinning nuclear magnetic resonance spectroscopy on H-USY (0.35 H+/Alf) and CD-HUSY (0.69 H+/Alf). These data demonstrate that the ubiquitous use of Alf sites as structural proxies for active H+ sites in zeolites can be imprecise, apparently because distorted Al structures that are not associated with acidic protons are sometimes detected as Alf sites. Monomolecular isobutane cracking and dehydrogenation rate constants, normalized non-rigorously by the number of Alf species, decreased with increasing Na+ content on both H-USY and CD-HUSY samples and became undetectable at sub-stoichiometric exchange levels (0.32 and 0.72 Na+/Alf ratios, respectively), an unexpected finding attributed incorrectly in previous studies to the presence of minority ‘‘super-acidic’’ sites. These rate constants, when normalized rigorously by the number of residual H+ sites were independent of Na+ content on both H-USY and CD-HUSY samples, reflecting the stoichiometric replacement of protons that are uniform in

  5. Acid strength of silica-supported oxide catalysts studied by microcalorimetric measurements of pyridine adsorption

    SciTech Connect

    Cardona-Martinez, N.; Dumesic, J.A. )

    1991-02-01

    Microcalorimetric measurements of the differential heat of pyridine adsorption were used to probe the distribution of acid strength on a series of silica-supported oxide catalysts. Depositing oxides of the following cations onto silica increased the acid strength of the catalyst: Ga{sup 3{plus}}, Zn{sup 2{plus}}, Al{sup 3{plus}}, Fe{sup 3{plus}}, Fe{sup 2{plus}}, Mg{sup 2{plus}}, and Sc{sup 3{plus}}. The acid strength distribution curves for the supported oxide samples showed either two or three regions of constant heat of adsorption while silica had an energetically homogeneous surface. The Ga, Al, and Sc samples were found to have both Bronsted and Lewis acidity while the remaining samples showed only Lewis acidity. Incremental adsorption of pyridine indicated that the initial region of highest heat corresponds to strong Lewis acidity while intermediate heats seemed to be due to weaker Lewis acid sites or a combination of Lewis and Bronsted acid sites. The final region of lowest heat was due to H-bonded pyridine on silica. Estimates of the entropies of adsorption were determined, providing information about the mobility of the adsorbed pyridine molecules. The initial differential heat of adsorption increases proportionally to the Sanderson electronegativity of the added oxide.

  6. Of Mental Models, Assumptions and Heuristics: The Case of Acids and Acid Strength

    ERIC Educational Resources Information Center

    McClary, LaKeisha Michelle

    2010-01-01

    This study explored what cognitive resources (i.e., units of knowledge necessary to learn) first-semester organic chemistry students used to make decisions about acid strength and how those resources guided the prediction, explanation and justification of trends in acid strength. We were specifically interested in the identifying and…

  7. The influence of dicarboxylic acids: Oxalic acid and tartaric acid on the compressive strength of glass ionomer cements

    NASA Astrophysics Data System (ADS)

    Permana, Ahmadi Jaya; Setyawati, Harsasi; Hamami, Murwani, Irmina Kris

    2016-03-01

    Glass ionomer cement (GIC) has limitation on the mechanical properties especially compressive strength. The change of compressive strength of GIC by adding oxalic acid and tartaric acid has been investigated. Oxalic acid and tartaric acid was added to the liquid components at concentrations of 0 - 15% (w/w). Powder component of GIC was made from optimum experimental powder glass SiO2-Al2O3-CaF2. GIC was characterized by compressive strength test, SEM-EDX and FTIR. The addition of tartaric acid to GIC has greater improvement than addition of oxalic acid. The addition of tartaric acid at 10 % (w/w) to GIC has greatest value of compressive strength.

  8. Polystyrene sulphonic acid resins with enhanced acid strength via macromolecular self-assembly within confined nanospace

    NASA Astrophysics Data System (ADS)

    Zhang, Xiaomin; Zhao, Yaopeng; Xu, Shutao; Yang, Yan; Liu, Jia; Wei, Yingxu; Yang, Qihua

    2014-01-01

    Tightening environmental legislation is driving the chemical industries to develop efficient solid acid catalysts to replace conventional mineral acids. Polystyrene sulphonic acid resins, as some of the most important solid acid catalysts, have been widely studied. However, the influence of the morphology on their acid strength—closely related to the catalytic activity—has seldom been reported. Herein, we demonstrate that the acid strength of polystyrene sulphonic acid resins can be adjusted through their reversible morphology transformation from aggregated to swelling state, mainly driven by the formation and breakage of hydrogen bond interactions among adjacent sulphonic acid groups within the confined nanospace of hollow silica nanospheres. The hybrid solid acid catalyst demonstrates high activity and selectivity in a series of important acid-catalysed reactions. This may offer an efficient strategy to fabricate hybrid solid acid catalysts for green chemical processes.

  9. Effects of acid diffusibility and affinity to cellulose on strength loss of polycarboxylic acid crosslinked fabrics.

    PubMed

    Ji, Bolin; Zhao, Cunyi; Yan, Kelu; Sun, Gang

    2016-06-25

    1,2,3,4-Butanetetracarboxylic acid (BTCA) imparts good anti-wrinkle property to cotton fabrics and results in significant strength loss due to cross-linking and acid degradation of cellulose simultaneously. However, benzophenone-3,3',4,4'- tetracarboxylic acid (BPTCA), an aromatic acid, crosslinks cellulose effectively but causes less strength loss to the products under similar conditions. The difference in damages to cellulose fibers was analyzed by using diffusibility and corresponding affinity of the acids to cellulose fibers, which were estimated by their molecular sizes and Hansen solubility parameters (HSP). Both experimental results and theoretical speculations revealed consistent agreement, indicating that smaller acid molecules could diffuse into cellulose fiber more rapidly and deeply, resulting in more acid degradation. Besides, the aliphatic acid such as BTCA has higher molecular affinity than BPTCA to cellulose, causing additional more degradation of cellulose. Both factors are potential reasons of the observed more severe tensile strength loss of the BTCA treated cotton fabrics. PMID:27083819

  10. Consequences of acid strength for isomerization and elimination catalysis on solid acids.

    PubMed

    Macht, Josef; Carr, Robert T; Iglesia, Enrique

    2009-05-13

    We address here the manner in which acid catalysis senses the strength of solid acids. Acid strengths for Keggin polyoxometalate (POM) clusters and zeolites, chosen because of their accurately known structures, are described rigorously by their deprotonation energies (DPE). Mechanistic interpretations of the measured dynamics of alkane isomerization and alkanol dehydration are used to obtain rate and equilibrium constants and energies for intermediates and transition states and to relate them to acid strength. n-Hexane isomerization rates were limited by isomerization of alkoxide intermediates on bifunctional metal-acid mixtures designed to maintain alkane-alkene equilibrium. Isomerization rate constants were normalized by the number of accessible protons, measured by titration with 2,6-di-tert-butylpyridine during catalysis. Equilibrium constants for alkoxides formed by protonation of n-hexene increased slightly with deprotonation energies (DPE), while isomerization rate constants decreased and activation barriers increased with increasing DPE, as also shown for alkanol dehydration reactions. These trends are consistent with thermochemical analyses of the transition states involved in isomerization and elimination steps. For all reactions, barriers increased by less than the concomitant increase in DPE upon changes in composition, because electrostatic stabilization of ion-pairs at the relevant transition states becomes more effective for weaker acids, as a result of their higher charge density at the anionic conjugate base. Alkoxide isomerization barriers were more sensitive to DPE than for elimination from H-bonded alkanols, the step that limits 2-butanol and 1-butanol dehydration rates; the latter two reactions showed similar DPE sensitivities, despite significant differences in their rates and activation barriers, indicating that slower reactions are not necessarily more sensitive to acid strength, but instead reflect the involvement of more unstable organic

  11. Bond strengths of all-ceramics: acid vs laser etching.

    PubMed

    Gökçe, B; Ozpinar, B; Dündar, M; Cömlekoglu, E; Sen, B H; Güngör, M A

    2007-01-01

    Various applications of dental lasers on dental materials have been proposed for surface modifications. This study evaluated whether laser etching could be an alternative to hydrofluoric acid (HF) etching. One hundred and ten lithia-based all-ceramic specimens (Empress 2) (R: 4 mm, h: 4 mm) were prepared and divided into five groups (n = 22/group). The untreated specimens served as the control, while one of the experimental groups was treated with 9.5% HF for 30 seconds. Three remaining test groups were treated with different laser (Er:YAG laser wavelength:2940 nm, OpusDent) power settings: 300 mJ, 600 mJ and 900 mJ. Ten specimens in each group were luted to the other 10 specimens by a dual-curing cement (Variolink II), and shear-bond strength (SBS) tests were performed (Autograph, crosshead speed: 0.5 mm/minute). The results were statistically analyzed (Kruskal Wallis and Mann Whitney-U, alpha = .05). Mean SBS (MPa) were 31.9 +/- 4.0, 41.4 +/- 4.3, 42.8 +/- 6.2, 29.2 +/- 4.5 and 27.4 +/- 3.8 for the control and HF, 300, 600 and 900 mJ groups, respectively. SEM evaluations revealed different surface morphologies depending on the laser parameters. The differences between HF acid and 300 mJ, when compared with the control, 600 and 900 mJ groups, were significant (p < .05). The 300 mJ laser group exhibited the highest shear-bond strength values, indicating that laser etching could also be used for surface treatments.

  12. Heuristic Reasoning in Chemistry: Making decisions about acid strength

    NASA Astrophysics Data System (ADS)

    McClary, LaKeisha; Talanquer, Vicente

    2011-07-01

    The characterization of students' reasoning strategies is of central importance in the development of instructional strategies that foster meaningful learning. In particular, the identification of shortcut reasoning procedures (heuristics) used by students to reduce cognitive load can help us devise strategies to facilitate the development of more analytical ways of thinking. The central goal of this qualitative study was thus to investigate heuristic reasoning as used by organic chemistry college students, focusing our attention on their ability to predict the relative acid strength of chemical compounds represented using explicit composition and structural features (i.e., structural formulas). Our results indicated that many study participants relied heavily on one or more of the following heuristics to make most of their decisions: reduction, representativeness, and lexicographic. Despite having visual access to reach structural information about the substances included in each ranking task, many students relied on isolated composition features to make their decisions. However, the specific characteristics of the tasks seemed to trigger heuristic reasoning in different ways. Although the use of heuristics allowed students to simplify some components of the ranking tasks and generate correct responses, it often led them astray. Very few study participants predicted the correct trends based on scientifically acceptable arguments. Our results suggest the need for instructional interventions that explicitly develop college chemistry students' abilities to monitor their thinking and evaluate the effectiveness of analytical versus heuristic reasoning strategies in different contexts.

  13. The Correlation of Binary Acid Strengths with Molecular Properties in First-Year Chemistry

    ERIC Educational Resources Information Center

    Fridgen, Travis D.

    2008-01-01

    This article deals with the rather complicated if not incorrect way that the strengths of binary acids are rationalized to students in many classrooms owing to the way it is presented in first-year chemistry textbooks. The common explanations, which use the homolytic bond dissociation energy as a rationalization of the trend in acid strengths when…

  14. Students' Understandings of Acid Strength: How Meaningful Is Reliability When Measuring Alternative Conceptions

    ERIC Educational Resources Information Center

    Bretz, Stacey Lowery; McClary, LaKeisha

    2015-01-01

    Most organic chemistry reactions occur by a mechanism that includes acid-base chemistry, so it is important that students develop and learn to use correct conceptions of acids and acid strength. Recent studies have described undergraduate organic chemistry students' cognitive resources related to the Brønsted-Lowry acid model and the Lewis acid…

  15. Support Effects on Bronsted acid site densities and alcohol dehydration turnover rates on tungsten oxide domains

    SciTech Connect

    Macht, Josef; Baertsch, Chelsey D.; May-Lozano, Marcos; Soled, Stuart L.; Wang, Yong; Iglesia, Enrique

    2005-03-01

    Initial activity and acid site density of several WAl, WSi (MCM41) and one WSn sample were determined. Trans/cis 2-butene selectivity is dependent on the support. Presumably, these differences are due to subtle differences in base strengths. 2-Butanol dehydration rates (per W-atom) reached maximum values at intermediate WOx surface densities on WAl, as reported for 2-butanol dehydration reactions on WZr. Titration results indicate that Bronsted acid sites are required for 2-butanol dehydration on WAl, WSi and WSn. UV-visible studies suggest that WAl is much more difficult to reduce than WZr. The detection of reduced centers on WAl, the number of which correlates to Bronsted acid site density and catalyst activity, as well as the temperature dependence of Bronsted acid site density indicate the in-situ formation of these active sites. We infer that this mechanism is common among all supported WOx samples described in this study. Turnover rates are a function of Bronsted acid site density only. High acid site densities lead to high turnover rates. Higher active site densities may cause stronger conjugate bases, as a higher electron density has to be stabilized, and thus weaker acidity, enabling a faster rate of product desorption. The maximum achievable active site density is dependent on the support. WZr reaches a higher active site density than WAl.

  16. Fracture Strength of Fused Silica From Photonic Signatures Around Collision Sites

    NASA Technical Reports Server (NTRS)

    Yost, William T.; Cramer, K Elliott

    2015-01-01

    Impact sites in glass affect its fracture strength. An analytical model that predicts fracture strength from grey-field polariscope (GFP) readings (photoelastic retardations) has been developed and reported in the literature. The model is suggestive that stress fields, resulting from impact damage, destablizes sites within the glass, which lead to pathways that cause strength degradation. Using data collected from fused silica specimens fabricated from outer window panes that were designed for the space shuttle, the model was tested against four categories of inflicted damage. The damage sites were cored from the window carcasses, examined with the GFP and broken using the ASTM Standard C1499-09 to measure the fracture strength. A correlation is made between the fracture strength and the photoelastic retardation measured at the damage site in each specimen. A least-squares fit is calculated. The results are compared with the predictions from the model. A plausible single-sided NDE damage site inspection method (a version of which is planned for glass inspection in the Orion Project) that relates photoelastic retardation in glass components to its fracture strength is presented.

  17. Emergence, Learning Difficulties, and Misconceptions in Chemistry Undergraduate Students' Conceptualizations of Acid Strength

    NASA Astrophysics Data System (ADS)

    Tümay, Halil

    2016-03-01

    Philosophical debates about chemistry have clarified that the issue of emergence plays a critical role in the epistemology and ontology of chemistry. In this article, it is argued that the issue of emergence has also significant implications for understanding learning difficulties and finding ways of addressing them in chemistry. Particularly, it is argued that many misconceptions in chemistry may derive from students' failure to consider emergence in a systemic manner by taking into account all relevant factors in conjunction. Based on this argument, undergraduate students' conceptions of acids, and acid strength (an emergent chemical property) were investigated and it was examined whether or not they conceptualized acid strength as an emergent chemical property. The participants were 41 third- and fourth-year undergraduate students. A concept test and semi-structured interviews were used to probe students' conceptualizations and reasoning about acid strength. Findings of the study revealed that the majority of the undergraduate students did not conceptualize acid strength as an emergent property that arises from interactions among multiple factors. They generally focused on a single factor to predict and explain acid strength, and their faulty responses stemmed from their failure to recognize and consider all factors that affect acid strength. Based on these findings and insights from philosophy of chemistry, promoting system thinking and epistemologically sound argumentative discourses among students is suggested for meaningful chemical education.

  18. Effect of soil acidity, soil strength and macropores on root growth and morphology of perennial grass species differing in acid-soil resistance.

    PubMed

    Haling, Rebecca E; Simpson, Richard J; Culvenor, Richard A; Lambers, Hans; Richardson, Alan E

    2011-03-01

    It is unclear whether roots of acid-soil resistant plants have significant advantages, compared with acid-soil sensitive genotypes, when growing in high-strength, acid soils or in acid soils where macropores may allow the effects of soil acidity and strength to be avoided. The responses of root growth and morphology to soil acidity, soil strength and macropores by seedlings of five perennial grass genotypes differing in acid-soil resistance were determined, and the interaction of soil acidity and strength for growth and morphology of roots was investigated. Soil acidity and strength altered root length and architecture, root hair development, and deformed the root tip, especially in acid-soil sensitive genotypes. Root length was restricted to some extent by soil acidity in all genotypes, but the adverse impact of soil acidity on root growth by acid-soil resistant genotypes was greater at high levels of soil strength. Roots reacted to soil acidity when growing in macropores, but elongation through high-strength soil was improved. Soil strength can confound the effect of acidity on root growth, with the sensitivity of acid-resistant genotypes being greater in high-strength soils. This highlights the need to select for genotypes that resist both acidity and high soil strength.

  19. Hydroxyalkylation of phenol to bisphenol F over heteropolyacid catalysts: The effect of catalyst acid strength on isomer distribution and kinetics.

    PubMed

    Wu, Xianzhang; Liu, Yutang; Liu, Ran; Wang, Longlu; Lu, Yanbing; Xia, Xinnian

    2016-11-01

    Hydroxyalkylation of phenol with formaldehyde to bisphenol F over heteropolyacid impregnated on clay was investigated. These catalysts displayed excellent catalytic performance for this reaction, especially that the effects of acid sites on the isomer distribution are obvious. Various solid catalysts were prepared by impregnating heteropolyacid on different kind of clay matrices, and their chemical compositions, textural properties, and acid strength of the heteropolyacid catalysts were characterized by EDX, BET, NH3-TPD, XRD, and FT-IR. Moreover, the effects of acid sites and reaction temperature on the yield and 4,4'-isomer distribution were launched by comparing the data obtained from the two kinds of catalysts. Furthermore, the kinetics of the hydroxyalkylation of phenol to BPF was established.

  20. Hydroxyalkylation of phenol to bisphenol F over heteropolyacid catalysts: The effect of catalyst acid strength on isomer distribution and kinetics.

    PubMed

    Wu, Xianzhang; Liu, Yutang; Liu, Ran; Wang, Longlu; Lu, Yanbing; Xia, Xinnian

    2016-11-01

    Hydroxyalkylation of phenol with formaldehyde to bisphenol F over heteropolyacid impregnated on clay was investigated. These catalysts displayed excellent catalytic performance for this reaction, especially that the effects of acid sites on the isomer distribution are obvious. Various solid catalysts were prepared by impregnating heteropolyacid on different kind of clay matrices, and their chemical compositions, textural properties, and acid strength of the heteropolyacid catalysts were characterized by EDX, BET, NH3-TPD, XRD, and FT-IR. Moreover, the effects of acid sites and reaction temperature on the yield and 4,4'-isomer distribution were launched by comparing the data obtained from the two kinds of catalysts. Furthermore, the kinetics of the hydroxyalkylation of phenol to BPF was established. PMID:27451037

  1. Shear bond strength of resin to acid/pumice-microabraded enamel.

    PubMed

    Royer, M A; Meiers, J C

    1995-01-01

    The effect of enamel microabrasion techniques consisting of either 18% hydrochloric acid in pumice or a commercially available abrasive/10% hydrochloric acid mixture, PREMA, on composite/enamel shear bond strengths was investigated. Sixty extracted third molars had the bonding surface flattened and were divided into six treatment groups (n=10) with the enamel treated prior to bonding as follows: Group 1-- untreated; Group 2--37% phosphoric acid etched for 30 seconds; Group 3--18% hydrochloric acid/pumice mixture applied for five 20-second treatments; Group 4--similar to Group 3 with additional 37% phosphoric acid etch; Group 5--treated with PREMA compound applied for five 20-second treatments; Group 6--similar to Group 5 treatment with additional 37% phosphoric acid. Herculite XR composite resin was then bonded to all samples using a VLC unit. Samples were tested in shear, and fractured enamel surfaces were evaluated using light microscopy to determine the enamel-to-resin failures. Resin bond strengths to microabraded and H3PO4-etched enamel were similar to bond strengths of untreated H3PO4-etched enamel and were significantly better than bond strengths to PREMA-treated or unetched enamel.

  2. Influence of hydrofluoric acid concentration on the flexural strength of a feldspathic ceramic.

    PubMed

    Venturini, Andressa B; Prochnow, Catina; May, Liliana G; Bottino, Marco C; Felipe Valandro, Luiz

    2015-08-01

    This study evaluated the effects of etching with increasing hydrofluoric (HF) acid concentrations on the roughness and flexural strength of a feldspathic ceramic. One hundred and fifty ceramic specimens (14×4×1.2 mm(2)) were produced from ceramic blocks (VitaBlocks Mark II). All specimens were polished, chamfered and sonically cleaned in isopropyl alcohol. Specimens were randomly divided into 5 groups (n=30): SC (control) no ceramic surface etching; HF1, HF3, HF5 and HF10 ceramic surface etching for 60s with 1%, 3%, 5% and 10% HF acid concentrations, respectively. Profilometry was performed in all specimens to evaluate roughness prior to flexural strength testing. Data were analyzed using one-way ANOVA and Tukey׳s test (α=0.05). Weibull module (m) and characteristic stress (σc) were also determined. HF acid etching, regardless of the concentration used, led to significantly rougher surfaces than the control (p<0.05). However, the mean flexural strength values were not statistically different among the etched groups (106.47 to 102.02 MPa). Acid etching significantly reduced the mean flexural strength when compared with the control (143.3 MPa). Weibull modulus of the groups was similar, except for the HF5 group that was higher compared to HF3. Flexural strength was similarly affected by the different HF acid concentrations tested, but roughness increased higher the acid concentration. Ceramic etching led to a significant reduction in strength when compared to the untreated ceramic, regardless of its concentration.

  3. Adverse experiences with nitric acid at the Savannah River Site

    SciTech Connect

    Durant, W.S.; Craig, D.K.; Vitacco, M.J.; McCormick, J.A.

    1991-06-01

    Nitric acid is used routinely at the Savannah River Site (SRS) in many processes. However, the site has experienced a number of adverse situations in handling nitric acid. These have ranged from minor injuries to personnel to significant explosions. This document compiles many of these events and includes discussions of process upsets, fires, injuries, and toxic effects of nitric acid and its decomposition products. The purpose of the publication is to apprise those using the acid that it is a potentially dangerous material and can react in many ways as demonstrated by SRS experience. 10 refs.

  4. The strength of the HIV-1 3' splice sites affects Rev function

    PubMed Central

    Kammler, Susanne; Otte, Marianne; Hauber, Ilona; Kjems, Jørgen; Hauber, Joachim; Schaal, Heiner

    2006-01-01

    Background The HIV-1 Rev protein is a key component in the early to late switch in HIV-1 splicing from early intronless (e.g. tat, rev) to late intron-containing Rev-dependent (e.g. gag, vif, env) transcripts. Previous results suggested that cis-acting sequences and inefficient 5' and 3' splice sites are a prerequisite for Rev function. However, we and other groups have shown that two of the HIV-1 5' splice sites, D1 and D4, are efficiently used in vitro and in vivo. Here, we focus on the efficiency of the HIV-1 3' splice sites taking into consideration to what extent their intrinsic efficiencies are modulated by their downstream cis-acting exonic sequences. Furthermore, we delineate their role in RNA stabilization and Rev function. Results In the presence of an efficient upstream 5' splice site the integrity of the 3' splice site is not essential for Rev function whereas an efficient 3' splice site impairs Rev function. The detrimental effect of a strong 3' splice site on the amount of Rev-dependent intron-containing HIV-1 glycoprotein coding (env) mRNA is not compensatable by weakening the strength of the upstream 5' splice site. Swapping the HIV-1 3' splice sites in an RRE-containing minigene, we found a 3' splice site usage which was variably dependent on the presence of the usual downstream exonic sequence. The most evident activation of 3' splice site usage by its usual downstream exonic sequence was observed for 3' splice site A1 which was turned from an intrinsic very weak 3' splice site into the most active 3' splice site, even abolishing Rev activity. Performing pull-down experiments with nuclear extracts of HeLa cells we identified a novel ASF/SF2-dependent exonic splicing enhancer (ESE) within HIV-1 exon 2 consisting of a heptameric sequence motif occurring twice (M1 and M2) within this short non-coding leader exon. Single point mutation of M1 within an infectious molecular clone is detrimental for HIV-1 exon 2 recognition without affecting Rev

  5. Splice Site Strength-Dependent Activity and Genetic Buffering by Poly-G Runs

    PubMed Central

    Xiao, Xinshu; Wang, Zefeng; Jang, Minyoung; Nutiu, Razvan; Wang, Eric T.; Burge, Christopher B.

    2009-01-01

    Pre-mRNA splicing is regulated through combinatorial activity of RNA motifs including splice sites and splicing regulatory elements (SREs). Here, we show that the activity of the G-run class of SREs is ∼4-fold higher when adjacent to intermediate strength 5'ss relative to weak 5'ss, and ∼1.3-fold higher relative to strong 5'ss. This dependence on 5'ss strength was observed in splicing reporters and in global microarray and mRNA-Seq analyses of splicing changes following RNAi against heterogeneous nuclear ribonucleoprotein (hnRNP) H, which crosslinked to G-runs adjacent to many regulated exons. An exon’s responsiveness to changes in hnRNP H levels therefore depends in a complex way on G-run abundance and 5'ss strength, and other splicing factors may function similarly. This pattern of activity enables G-runs and hnRNP H to buffer the effects of 5'ss mutations, augmenting the frequency of 5'ss polymorphism and the evolution of new splicing patterns. PMID:19749754

  6. Hydrogen trapping sites in high-strength steels at delayed fracture

    SciTech Connect

    Takai, Kenichi; Seki, Junichi; Homma, Yoshikazu

    1996-12-01

    Hydrogen trapping sites in high-strength steels hardened by cold drawing (PC Wire) and heat treatment (PC Bar) were investigated using thermal desorption spectroscopy (TDS) and secondary ion mass spectrometry (SIMS). Both steels were stressed and dipped in 20% NH{sub 4}SCN solution at 323 K to occlude hydrogen for TDS analysis, whereas they were stressed and dipped in D{sub 2}O and 20% NH{sub 4}SCN solution at 323 K to occlude deuterium for SIMS analysis. TDS analysis shows that PC Wire occludes hydrogen at the peaks of 470 K and 630 K, whereas PC Bar mainly occludes hydrogen at 470 K and slightly at 630 K. SIMS analysis makes it possible to observe the trapping sites of deuterium instead of hydrogen in high-strength steels. PC Wire traps deuterium along the drawing direction. PC Bar traps deuterium at the grain boundaries and on the inclusions. The correspondence between TDS and SIMS shows that hydrogen at 470 K is evolved from the matrix that includes dislocations and grain boundaries. In contrast, hydrogen at 630 K is evolved from inclusions and segregation of P.

  7. The effect of elevated temperature on the strength parameters of silica acid ester consolidated porous limestones

    NASA Astrophysics Data System (ADS)

    Pápay, Zita; Török, Ákos

    2013-04-01

    The porous limestone is one of the most widespread construction materials of the monuments in Central Europe, with emblematic buildings in Vienna, Bratislava, Budapest and many other cities of Austria, Slovakia, Czech Republic and Hungary. The restoration of these monuments very often requires the consolidation of the porous limestone material, where various types of consolidants are used to strengthen the highly weathered stone. Our research focused on the understanding of the behaviour of consolidated porous limestone when the material is subjected to higher temperatures. Test procedure included the preparation of cylindrical test specimens from the Miocene porous limestone which was followed by consolidation by four various types of silica acid ester. The samples after consolidation were heated to 300 and 600 °C in electric oven. The material properties such as ultrasonic pulse velocity, density were tested before and after the treatment. Indirect tensile strength (Brazilian test) was used to compare the strength parameters of non treated and consolidated samples. Silica acid ester treated samples after heating were also measured in terms of strength, density and ultrasonic pulse velocity. The results show that there are significant changes in strength of various pre-treated samples after heating indicating the sensitivity of the materials to temperature changes and accidental fire.

  8. Catalytic Consequences of Acid Strength in the Conversion of Methanol to Dimethyl Ether

    SciTech Connect

    Carr, Robert T.; Neurock, Matthew; Iglesia, Enrique

    2011-02-14

    The effects of acid identity on CH{sub 3}OH dehydration are examined here using density functional theory (DFT) estimates of acid strength (as deprotonation energies, DPE) and reaction energies, combined with rate data on Keggin polyoxometalate (POM) clusters and zeolite H-BEA. Measured first-order (k{sub mono}) and zero-order (k{sub dimer}) CH3OH dehydration rate constants depend exponentially on DPE for POM clusters; the value of k{sub mono} depends more strongly on DPE than k{sub dimer} does. The chemical significance of these rate parameters and the basis for their dependences on acid strength were established by using DFT to estimate the energies of intermediates and transition states involved in elementary steps that are consistent with measured rate equations. We conclude from this treatment that CH{sub 3}OH dehydration proceeds via direct reactions of co-adsorbed CH{sub 3}OH molecules for relevant solid acids and reaction conditions. Methyl cations formed at ion-pair transition states in these direct routes are solvated by H{sub 2}O and CH{sub 3}OH more effectively than those in alternate sequential routes involving methoxide formation and subsequent reaction with CH{sub 3}OH. The stability of ion-pairs, prevalent as intermediates and transition states on solid acids, depends sensitively on DPE because of concomitant correlations between the stability of the conjugate anionic cluster and DPE. The chemical interpretation of k{sub mono} and k{sub dimer} from mechanism-based rate equations, together with thermochemical cycles of their respective transition state formations, show that similar charge distributions in the intermediate and transition state involved in k{sub dimer} cause its weaker dependence on DPE. Values of k{sub mono} involve uncharged reactants and the same ion-pair transition state as k{sub dimer}; these species sense acid strength differently and cause the larger effects of DPE on k{sub mono}. Confinement effects in H-BEA affect the value of

  9. Mutations of fumarase that distinguish between the active site and a nearby dicarboxylic acid binding site.

    PubMed Central

    Weaver, T.; Lees, M.; Banaszak, L.

    1997-01-01

    Two mutant forms of fumarase C from E. coli have been made using PCR and recombinant DNA. The recombinant form of the protein included a histidine arm on the C-terminal facilitating purification. Based on earlier studies, two different carboxylic acid binding sites, labeled A- and B-, were observed in crystal structures of the wild type and inhibited forms of the enzyme. A histidine at each of the sites was mutated to an asparagine. H188N at the A-site resulted in a large decrease in specific activity, while the H129N mutation at the B-site had essentially no effect. From the results, we conclude that the A-site is indeed the active site, and a dual role for H188 as a potential catalytic base is proposed. Crystal structures of the two mutant proteins produced some unexpected results. Both mutations reduced the affinity for the carboxylic acids at their respective sites. The H129N mutant should be particularly useful in future kinetic studies because it sterically blocks the B-site with the carboxyamide of asparagine assuming the position of the ligand's carboxylate. In the H188N mutation at the active site, the new asparagine side chain still interacts with an active site water that appears to have moved slightly as a result of the mutation. PMID:9098893

  10. Evaluating Variability and Uncertainty of Geological Strength Index at a Specific Site

    NASA Astrophysics Data System (ADS)

    Wang, Yu; Aladejare, Adeyemi Emman

    2016-09-01

    Geological Strength Index (GSI) is an important parameter for estimating rock mass properties. GSI can be estimated from quantitative GSI chart, as an alternative to the direct observational method which requires vast geological experience of rock. GSI chart was developed from past observations and engineering experience, with either empiricism or some theoretical simplifications. The GSI chart thereby contains model uncertainty which arises from its development. The presence of such model uncertainty affects the GSI estimated from GSI chart at a specific site; it is, therefore, imperative to quantify and incorporate the model uncertainty during GSI estimation from the GSI chart. A major challenge for quantifying the GSI chart model uncertainty is a lack of the original datasets that have been used to develop the GSI chart, since the GSI chart was developed from past experience without referring to specific datasets. This paper intends to tackle this problem by developing a Bayesian approach for quantifying the model uncertainty in GSI chart when using it to estimate GSI at a specific site. The model uncertainty in the GSI chart and the inherent spatial variability in GSI are modeled explicitly in the Bayesian approach. The Bayesian approach generates equivalent samples of GSI from the integrated knowledge of GSI chart, prior knowledge and observation data available from site investigation. Equations are derived for the Bayesian approach, and the proposed approach is illustrated using data from a drill and blast tunnel project. The proposed approach effectively tackles the problem of how to quantify the model uncertainty that arises from using GSI chart for characterization of site-specific GSI in a transparent manner.

  11. Synthesis of kojic acid derivatives as secondary binding site probes of D-amino acid oxidase

    PubMed Central

    Raje, Mithun; Hin, Niyada; Duvall, Bridget; Ferraris, Dana V.; Berry, James F.; Thomas, Ajit G.; Alt, Jesse; Rojas, Camilo; Slusher, Barbara S.; Tsukamoto, Takashi

    2013-01-01

    A series of kojic acid (5-hydroxy-2-hydroxymethyl-4H-pyran-4-one) derivatives were synthesized and tested for their ability to inhibit D-amino acid oxidase (DAAO). Various substituents were incorporated into kojic acid at its 2-hydroxymethyl group. These analogs serve as useful molecular probes to explore the secondary binding site, which can be exploited in designing more potent DAAO inhibitors. PMID:23683589

  12. Active and Passive Application of the Phosphoric Acid on the Bond Strength of Lithium Disilicate.

    PubMed

    Giraldo, Tatiana Cardona; Villada, Vanessa Roldan; Castillo, Mauricio Peña; Gomes, Osnara Maria Mongruel; Bittencourt, Bruna Fortes; Dominguez, John Alexis

    2016-01-01

    The objective of this study was to evaluate the effect of passive or active phosphoric acid (PA) application after hydrofluoric acid (HA) treatment on the microshear bond strength of lithium disilicate. Thirty ceramic discs were made with IPS Emax 2 (10 mm thick and 10 mm diameter). The specimens were divided into 3 groups, A: 9.6% HA application; AF: 9.6% HA application + cleaning with 37% PA in passive mode and AFF: 9.6% HA application + cleaning with 37% PA in active mode. For the microshear test, four tygons (0.9 mm diameter and 0.2 mm high) were filled with resin cement (RelyX Ultimate) and placed on the ceramic disks. After testing, the fracture modes were examined under scanning electron microscopy. Data were analyzed by one-way ANOVA and Tukey's post test (α=0.05). The bond strength values were significantly higher in Group AFF (11.0±2.5 MPa) compared with group A (8.1±2.6 MPa) (p<0.002). AF group was not statistically different (9.4±2.5 MPa) from Group A. It was concluded that the active application of 37% PA after 9.6% HA increases the microshear bond strength values between the resin cement and lithium disilicate ceramic.

  13. Dehydration of lactic acid to acrylic acid over lanthanum phosphate catalysts: the role of Lewis acid sites.

    PubMed

    Guo, Zhen; Theng, De Sheng; Tang, Karen Yuanting; Zhang, Lili; Huang, Lin; Borgna, Armando; Wang, Chuan

    2016-09-14

    Lanthanum phosphate (LaP) nano-rods were synthesized using n-butylamine as a shape-directing agent (SDA). The resulting catalysts were applied in the dehydration of lactic acid to acrylic acid. Aiming to understand the nature of the active sites, the chemical and physical properties of LaP materials were studied using a variety of characterization techniques. This study showed that the SDA not only affected the porosity of the LaP materials but also modified the acid-base properties. Clearly, the modification of the acid-base properties played a more critical role in determining the catalytic performance than porosity. An optimized catalytic performance was obtained on the LaP catalyst with a higher concentration of Lewis acid sites. Basic sites showed negative effects on the stability of the catalysts. Good stability was achieved when the catalyst was prepared using the appropriate SDA/La ratio. PMID:27514871

  14. Dehydration of lactic acid to acrylic acid over lanthanum phosphate catalysts: the role of Lewis acid sites.

    PubMed

    Guo, Zhen; Theng, De Sheng; Tang, Karen Yuanting; Zhang, Lili; Huang, Lin; Borgna, Armando; Wang, Chuan

    2016-09-14

    Lanthanum phosphate (LaP) nano-rods were synthesized using n-butylamine as a shape-directing agent (SDA). The resulting catalysts were applied in the dehydration of lactic acid to acrylic acid. Aiming to understand the nature of the active sites, the chemical and physical properties of LaP materials were studied using a variety of characterization techniques. This study showed that the SDA not only affected the porosity of the LaP materials but also modified the acid-base properties. Clearly, the modification of the acid-base properties played a more critical role in determining the catalytic performance than porosity. An optimized catalytic performance was obtained on the LaP catalyst with a higher concentration of Lewis acid sites. Basic sites showed negative effects on the stability of the catalysts. Good stability was achieved when the catalyst was prepared using the appropriate SDA/La ratio.

  15. Shear bond strength of resin cement to an acid etched and a laser irradiated ceramic surface

    PubMed Central

    Motro, Pelin Fatma Karagoz; Yurdaguven, Haktan

    2013-01-01

    PURPOSE To evaluate the effects of hydrofluoric acid etching and Er,Cr:YSGG laser irradiation on the shear bond strength of resin cement to lithium disilicate ceramic. MATERIALS AND METHODS Fifty-five ceramic blocks (5 mm × 5 mm × 2 mm) were fabricated and embedded in acrylic resin. Their surfaces were finished with 1000-grit silicon carbide paper. The blocks were assigned to five groups: 1) 9.5% hydrofluoric-acid etching for 60 s; 2-4), 1.5-, 2.5-, and 6-W Er,Cr:YSGG laser applications for 60 seconds, respectively; and 5) no treatment (control). One specimen from each group was examined using scanning electron microscopy. Ceramic primer (Rely X ceramic primer) and adhesive (Adper Single Bond) were applied to the ceramic surfaces, followed by resin cement to bond the composite cylinders, and light curing. Bonded specimens were stored in distilled water at 37℃ for 24 hours. Shear bond strengths were determined by a universal testing machine at 1 mm/min crosshead speed. Data were analyzed using Kruskal-Wallis and Mann-Whitney U-tests (α=0.05). RESULTS Adhesion was significantly stronger in Group 2 (3.88 ± 1.94 MPa) and Group 3 (3.65 ± 1.87 MPa) than in Control group (1.95 ± 1.06 MPa), in which bonding values were lowest (P<.01). No significant difference was observed between Group 4 (3.59 ± 1.19 MPa) and Control group. Shear bond strength was highest in Group 1 (8.42 ± 1.86 MPa; P<.01). CONCLUSION Er,Cr:YSGG laser irradiation at 1.5 and 2.5 W increased shear bond strengths between ceramic and resin cement compared with untreated ceramic surfaces. Irradiation at 6 W may not be an efficient ceramic surface treatment technique. PMID:23755333

  16. Acid aerosol measurements at a suburban Connecticut site

    NASA Astrophysics Data System (ADS)

    Keeler, Gerald J.; Spengler, John D.; Castillo, Raymond A.

    Atmospheric acidity data were gathered during a year-long field project investigating the possible health effects of acid aerosol in a rural community in southwestern Connecticut. This site was chosen because the air quality is frequently influenced by pollutants transported from the New York-New Jersey corridor as well as from the Midwest U.S. An annular denuder filter-pack system utilized to obtain daily measurements of gaseous HNO 3, HONO, SO 2, and NH 3; plus fine particle SO 42-, NO 3-, and H +. Fine particle mass ( d ⩽ 2.1 μm) and PM10 (particles d ⩽ 10 μm) were also measured. Ozone concentrations and basic meteorological data were also obtained continuously. The atmosphere was acidic with average concentrations of HONO (16 nmol m -3), HNO 3 (42 nmol m -3), and H + (42 nmol m -3), observed from May to September 1988. Atmospheric ammonia concentrations were fairly low averaging 34 nmol m -3 during the same period, and suggesting the neutralizing capacity of the air was significant to neutralize all the acidic species present. Neutralization of acidic particles by reactions on the filter media after collection resulted in a loss of approximately 10% of the measured particle strong acidity for the summertime period investigated. Concentrations of ozone and acidic gases tended to peak with mixed layer flow from the south-southwest while particulate acidity was highest with flow predominantly from the west-soutwest. Hourly ozone concentrations greater than 100 ppb were observed on 31 different days during the monitoring, and concentrations greater than 150 ppb measured on 14 days. HNO 3 and aerosol strong acidity (H +) concentrations reached 174 and 199 nmol m -3, respectively during the summer months.

  17. Phosphoric acid-etching promotes bond strength and formation of acid-base resistant zone on enamel.

    PubMed

    Li, N; Nikaido, T; Alireza, S; Takagaki, T; Chen, J-H; Tagami, J

    2013-01-01

    This study examined the effect of phosphoric acid (PA) etching on the bond strength and acid-base resistant zone (ABRZ) formation of a two-step self-etching adhesive (SEA) system to enamel. An etch-and-rinse adhesive (EAR) system Single Bond (SB) and a two-step SEA system Clearfil SE Bond (SE) were used. Human teeth were randomly divided into four groups according to different adhesive treatments: 1) SB; 2) SE; 3) 35% PA etching→SE primer→SE adhesive (PA/SEp+a); (4) 35% PA etching→SE adhesive (PA/SEa). Microshear bond strength to enamel was measured and then statistically analyzed using one-way analysis of variance and the Tukey honestly significant difference test. The failure mode was recorded and analyzed by χ( 2 ) test. The etching pattern of the enamel surface was observed with scanning electron microscope (SEM). The bonded interface was exposed to a demineralizing solution (pH=4.5) for 4.5 hours and then 5% sodium hypochlorite with ultrasonication for 30 minutes. After argon-ion etching, the interfacial ultrastructure was observed using SEM. The microshear bond strength to enamel of the SE group was significantly lower (p<0.05) than that of the three PA-etched groups, although the latter three were not significantly different from one another. The ABRZ was detected in all the groups. In morphological observation, the ABRZ in the three PA-etched groups were obviously thicker compared with the SE group with an irregular wave-shaped edge.

  18. Investigation of the ionic strength dependence of Ulva lactuca acid functional group pK(a)s by manual alkalimetric titrations.

    PubMed

    Schijf, Johan; Ebling, Alina M

    2010-03-01

    We performed a series of manual alkalimetric titrations in NaCl solutions (0.01-5.0 M) at T = 25 degrees C on both fresh and dehydrated samples of the marine chlorophyte Ulva lactuca (sea lettuce), a strong metal accumulator holding considerable promise in biosorbent and biomonitor applications. Functional groups were characterized in terms of their number, site densities, and acid dissociation constants (pK(a)s). FITEQL4.0 modeling shows that, at any ionic strength, titration curves for dehydrated biomass in the pH range 2-10 are adequately described by three functional groups with remarkably uniform site densities of about 5 x 10(-4) mol/g. Lower site densities for fresh U. lactuca are consistent with approximately 87% water content. The pK(a)s display pronounced ionic strength dependent behavior obeying an extended Debye-Huckel relation. Extrapolation to I = 0 yields values of 4.26 +/- 0.04, 6.44 +/- 0.02, and 9.56 +/- 0.04. This information by itself is insufficient to unambiguously identify the groups. Similar site densities suggest that all three are linked to major molecular building blocks of the cell material, pointing to carboxylic acids, phosphate esters, and amines as likely candidates. Highly acidic sulfate esters, not detected in our titrations, may also play a role in trace metal adsorption on U. lactuca.

  19. Computational studies of Bronsted acid sites in zeolites

    SciTech Connect

    Curtiss, L.A.; Iton, L.E.; Zygmunt, S.A.

    1995-01-01

    The authors have performed high-level ab initio calculations using both Hartree-Fock (HF) and Moller-Plesset perturbation theory (MP2) to study the geometry and energetics of the adsorption complex involving H{sub 2}O and the Bronsted acid site in the zeolite H-ZSM-5. In these calculations, which use aluminosilicate cluster models for the zeolite framework with as many a 28 T atoms (T = Si, Al), we included geometry optimization in the local vicinity of the acid site at the HF/6-31G(d) level of theory, and have calculated corrections for zero-point energies, extensions for zero-point energies, extensions to higher basis sets, and the influence of electron correlation. Results for the adsorption energy and geometry of this complex are reported and compared with previous theoretical and experimental values.

  20. Correlation of zeolite ZSM-5 acid strength with catalytic activity and selectivity. Final report. [Cobalt and cobalt/thorium impregnated H-ZSM-5

    SciTech Connect

    Hayhurst, D.T.

    1984-04-01

    Isosteric heats of ammonia adsorption were determined for several cobalt and cobalt/thorium impregnated H-ZSM-5. Samples were impregnated to the 3, 6 and 9 wt % levels using an incipient wetness technique and either acetone or water for dissolving the metal nitrate salt. The metal impregnation resulted in a lowering of the maximum heat of ammonia adsorption and in a corresponding lowering of the maximum acid site strength. The ammonia adsorption capacity and adsorption heat was unaltered by a reduction of the metal-impregnated samples in hydrogen, indicating that the incipient wetness method leads to cobalt which was most likely ion-exchanged into the zeolite rather than impregnated into the sieve in a reducable form. In addition, the impregnated cobalt caused a slight steric hinderance to the diffusion of n-butane into the metal-loaded zeolite. The use of either water or acetone for dissolving the metal nitrate salt altered the acid-strength distribution. Water was found to lower the acid-site strength to a greater degree than the acetone. 11 references, 8 figures.

  1. Multi-site study of diffusion metric variability: effects of site, vendor, field strength, and echo time on regions-of-interest and histogram-bin analyses

    NASA Astrophysics Data System (ADS)

    Helmer, K. G.; Chou, M.-C.; Preciado, R. I.; Gimi, B.; Rollins, N. K.; Song, A.; Turner, J.; Mori, S.

    2016-03-01

    It is now common for magnetic-resonance-imaging (MRI) based multi-site trials to include diffusion-weighted imaging (DWI) as part of the protocol. It is also common for these sites to possess MR scanners of different manufacturers, different software and hardware, and different software licenses. These differences mean that scanners may not be able to acquire data with the same number of gradient amplitude values and number of available gradient directions. Variability can also occur in achievable b-values and minimum echo times. The challenge of a multi-site study then, is to create a common protocol by understanding and then minimizing the effects of scanner variability and identifying reliable and accurate diffusion metrics. This study describes the effect of site, scanner vendor, field strength, and TE on two diffusion metrics: the first moment of the diffusion tensor field (mean diffusivity, MD), and the fractional anisotropy (FA) using two common analyses (region-of-interest and mean-bin value of whole brain histograms). The goal of the study was to identify sources of variability in diffusion-sensitized imaging and their influence on commonly reported metrics. The results demonstrate that the site, vendor, field strength, and echo time all contribute to variability in FA and MD, though to different extent. We conclude that characterization of the variability of DTI metrics due to site, vendor, field strength, and echo time is a worthwhile step in the construction of multi-center trials.

  2. Analysis of the strengths and weaknesses of acid rain electronic data reports

    SciTech Connect

    Schott, J.

    1997-12-31

    Entergy Corporation is a Phase II utility with a fossil generation base composed primarily of natural gas and low sulfur coal. This paper presents an analysis of a large Phase II utility`s continuous emissions monitoring data reported to EPA under Title IV Acid Rain. Electric utilities currently report hourly emissions of NO{sub x}, SO{sub 2}, CO{sub 2}, fuel use, and generation through electronic data reports to EPA. This paper describes strengths and weaknesses of the data reported to EPA as determined through an analysis of 1995 data. Emissions reported by this company under acid rain for SO{sub 2} and NO{sub x} are very different from emissions reported to state agencies for annual emission inventory purposes in past years and will represent a significant break with historic trends. A comparison of emissions has been made of 1995 emissions reported under Electronic Data Reports to the emissions that would have been reported using emission factors and fuel data in past years. In addition, the paper examines the impacts of 40 CFR Part 75 Acid Rain requirements such as missing data substitution and monitor bias adjustments. Measurement system errors including stack flow measurement and false NO{sub x}Lb/MMBtu readings at very low loads are discussed. This paper describes the implications for public policy, compliance, emissions inventories, and business decisions of Part 75 acid rain monitoring and reporting requirements.

  3. Dissolution of Simulated and Radioactive Savannah River Site High-Level Waste Sludges with Oxalic Acid & Citric Acid Solutions

    SciTech Connect

    STALLINGS, MARY

    2004-07-08

    This report presents findings from tests investigating the dissolution of simulated and radioactive Savannah River Site sludges with 4 per cent oxalic acid and mixtures of oxalic and citric acid previously recommended by a Russian team from the Khlopin Radium Institute and the Mining and Chemical Combine (MCC). Testing also included characterization of the simulated and radioactive waste sludges. Testing results showed the following: Dissolution of simulated HM and PUREX sludges with oxalic and citric acid mixtures at SRTC confirmed general trends reported previously by Russian testing. Unlike the previous Russian testing six sequential contacts of a mixture of oxalic acid citric acids at a 2:1 ratio (v/w) of acid to sludge did not produce complete dissolution of simulated HM and PUREX sludges. We observed that increased sludge dissolution occurred at a higher acid to sludge ratio, 50:1 (v/w), compared to the recommended ratio of 2:1 (v/w). We observed much lower dissolution of aluminum in a simulated HM sludge by sodium hydroxide leaching. We attribute the low aluminum dissolution in caustic to the high fraction of boehmite present in the simulated sludge. Dissolution of HLW sludges with 4 per cent oxalic acid and oxalic/citric acid followed general trends observed with simulated sludges. The limited testing suggests that a mixture of oxalic and citric acids is more efficient for dissolving HM and PUREX sludges and provides a more homogeneous dissolution of HM sludge than oxalic acid alone. Dissolution of HLW sludges in oxalic and oxalic/citric acid mixtures produced residual sludge solids that measured at higher neutron poison to equivalent 235U weight ratios than that in the untreated sludge solids. This finding suggests that residual solids do not present an increased nuclear criticality safety risk. Generally the neutron poison to equivalent 235U weight ratios of the acid solutions containing dissolved sludge components are lower than those in the untreated

  4. Acidity of edge surface sites of montmorillonite and kaolinite

    NASA Astrophysics Data System (ADS)

    Liu, Xiandong; Lu, Xiancai; Sprik, Michiel; Cheng, Jun; Meijer, Evert Jan; Wang, Rucheng

    2013-09-01

    Acid-base chemistry of clay minerals is central to their interfacial properties, but up to now a quantitative understanding on the surface acidity is still lacking. In this study, with first principles molecular dynamics (FPMD) based vertical energy gap technique, we calculate the acidity constants of surface groups on (0 1 0)-type edges of montmorillonite and kaolinite, which are representatives of 2:1 and 1:1-type clay minerals, respectively. It shows that tbnd Si-OH and tbnd Al-OH2OH groups of kaolinite have pKas of 6.9 and 5.7 and those of montmorillonite have pKas of 7.0 and 8.3, respectively. For each mineral, the calculated pKas are consistent with the experimental ranges derived from fittings of titration curves, indicating that tbnd Si-OH and tbnd Al-OH2OH groups are the major acidic sites responsible to pH-dependent experimental observations. The effect of Mg substitution in montmorillonite is investigated and it is found that Mg substitution increases the pKas of the neighboring tbnd Si-OH and tbnd Si-OH2 groups by 2-3 pKa units. Furthermore, our calculation shows that the pKa of edge tbnd Mg-(OH2)2 is as high as 13.2, indicating the protonated state dominates under common pH. Together with previous adsorption experiments, our derived acidity constants suggest that tbnd Si-O- and tbnd Al-(OH)2 groups are the most probable edge sites for complexing heavy metal cations.

  5. Development and Assessment of a Diagnostic Tool to Identify Organic Chemistry Students' Alternative Conceptions Related to Acid Strength

    ERIC Educational Resources Information Center

    McClary, LaKeisha M.; Bretz, Stacey Lowery

    2012-01-01

    The central goal of this study was to create a new diagnostic tool to identify organic chemistry students' alternative conceptions related to acid strength. Twenty years of research on secondary and college students' conceptions about acids and bases has shown that these important concepts are difficult for students to apply to qualitative problem…

  6. Effective charge on acetylcholinesterase active sites determined from the ionic strength dependence of association rate constants with cationic ligands.

    PubMed

    Nolte, H J; Rosenberry, T L; Neumann, E

    1980-08-01

    The reaction of the specific fluorescent cationic ligand N-methylacridinium with the active site of 11S acetylcholinesterase from electric eel was monitored by temperature-jump relaxation kinetics at a variety of ionic strengths. The ionic strength dependence of the bimolecular association rate constant is analyzed with a Brønsted-Debye-Hückel expression and leads to estimates of the association rate constant at zero ionic strength of K120 = 1.1 X 10(10) M-1 S-1 at 25 degrees C and the net charge number of the enzyme active site of ZE = -6.3. The ionic strength dependence of the second-order hydrolysis rate constant kcat/Kapp for acetylthiocholine under steady-state conditions is also very pronounced and indicates a value of ZE = -9. Thus, a large effective negative charge on the enzyme active site appears to be a general characteristic of its interaction with cationic ligands. The ionic strength dependence of Kcat/Kapp is identical with that of sodium chloride, sodium phosphate, and sodium citrate, thus ruling out any possibility that the phenomena arise from a specific, partially competitive binding of Na+ to the enzyme active site. Substitution of the calculated electrostatic parameters into theoretical equations indicates that the most significant effect of these ZE values is a 2-3 order of magnitude reduction in the rate constant for dissociation of the initial ligand-enzyme encounter complex; this decrease renders the bimolecular reaction diffusion controlled. The high value of k120 and the space requirements of six to nine charged groups suggest that regions of the enzyme surface area larger than the catalytic sites themselves are effective in trapping cationic ligands.

  7. Comparison of bond strength and surface morphology of dental enamel for acid and Nd-YAG laser etching

    NASA Astrophysics Data System (ADS)

    Parmeswearan, Diagaradjane; Ganesan, Singaravelu; Ratna, P.; Koteeswaran, D.

    1999-05-01

    Recently, laser pretreatment of dental enamel has emerged as a new technique in the field of orthodontics. However, the changes in the morphology of the enamel surface is very much dependent on the wavelength of laser, emission mode of the laser, energy density, exposure time and the nature of the substance absorbing the energy. Based on these, we made a comparative in vitro study on laser etching with acid etching with reference to their bond strength. Studies were conducted on 90 freshly extracted, non carious, human maxillary or mandibular anteriors and premolars. Out of 90, 60 were randomly selected for laser irradiation. The other 30 were used for conventional acid pretreatment. The group of 60 were subjected to Nd-YAG laser exposure (1060 nm, 10 Hz) at differetn fluences. The remaining 30 were acid pretreated with 30% orthophosphoric acid. Suitable Begg's brackets were selected and bound to the pretreated surface and the bond strength were tested using Instron testing machine. The bond strength achieved through acid pretreatment is found to be appreciably greater than the laser pretreated tooth. Though the bond strength achieved through the acid pretreated tooth is found to be significantly greater than the laser pretreated specimens, the laser pretreatement is found to be successful enough to produce a clinically acceptable bond strength of > 0.60 Kb/mm. Examination of the laser pre-treated tooth under SEM showed globule formation which may produce the mechanical interface required for the retention of the resin material.

  8. A computer program for geochemical analysis of acid-rain and other low-ionic-strength, acidic waters

    USGS Publications Warehouse

    Johnsson, P.A.; Lord, D.G.

    1987-01-01

    ARCHEM, a computer program written in FORTRAN 77, is designed primarily for use in the routine geochemical interpretation of low-ionic-strength, acidic waters. On the basis of chemical analyses of the water, and either laboratory or field determinations of pH, temperature, and dissolved oxygen, the program calculates the equilibrium distribution of major inorganic aqueous species and of inorganic aluminum complexes. The concentration of the organic anion is estimated from the dissolved organic concentration. Ionic ferrous iron is calculated from the dissolved oxygen concentration. Ionic balances and comparisons of computed with measured specific conductances are performed as checks on the analytical accuracy of chemical analyses. ARCHEM may be tailored easily to fit different sampling protocols, and may be run on multiple sample analyses. (Author 's abstract)

  9. Program Strengths and Opportunities for Improvement Identified by Residents During ACGME Site Visits in 5 Surgical Specialties.

    PubMed

    Caniano, Donna A; Hamstra, Stanley J

    2016-05-01

    Background There is limited information about how residents in surgical specialties view program strengths and opportunities for improvement (OFIs). Objective This study aggregated surgical residents' perspectives on program strengths and OFIs to determine whether there was agreement in perspectives among residents in 5 surgical specialties. Methods Resident consensus lists of program strengths and areas for improvement were aggregated from site visits reports during 2012 and 2013 for obstetrics and gynecology, orthopaedic surgery, otolaryngology, plastic surgery, and surgery programs. Four trained individuals coded each strength or OFI in 1 of 3 categories: (1) factors common to all specialties; (2) program or institutional resources; and (3) factors unique to surgical specialties. Themes were classified as most frequent when listed by residents in more than 20% of the programs and less frequent when listed by residents in less than 20% of the programs. Results This study included a total of 359 programs, representing 27% to 49% of the Accreditation Council for Graduate Medical Education accredited programs in the 5 specialties. The most frequent strengths were progressive autonomy, collegiality, program leadership, and operative volume. Improving research and didactics, increasing faculty teaching and attendance at educational sessions, and increasing the number of nurse practitioners and physician assistants were common OFIs. Conclusions Factors identified as important by surgical residents related to their learning environment, their educational program, and program and institutional support. Across programs in the study, similar attributes were listed as both program strengths and OFIs. PMID:27168889

  10. Serum Uric Acid Is Positively Associated with Handgrip Strength among Japanese Community-Dwelling Elderly Women.

    PubMed

    Kawamoto, Ryuichi; Ninomiya, Daisuke; Kasai, Yoshihisa; Kusunoki, Tomo; Ohtsuka, Nobuyuki; Kumagi, Teru; Abe, Masanori

    2016-01-01

    Serum uric acid (UA) has strong anti-oxidant properties. Muscle strength and mass decrease with age, and recently, this decrease has been defined as sarcopenia. Sarcopenia may be triggered by oxidative stress. We investigated whether serum UA is associated with handgrip strength (HGS), which is a useful indicator of sarcopenia, among Japanese community-dwelling elderly persons. The present study included 602 men aged 72 ± 7 years and 847 women aged 71 ± 6 years from a rural village. We examined the cross-sectional relationship between serum UA and HGS. In both genders, HGS increased significantly with increased serum UA levels. A multiple linear regression analysis using HGS as an objective variable and various confounding factors as explanatory variables showed that in men age, drinking status, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and estimated glomerular filtration ratio (eGFRCKDEPI) were independently and significantly associated with HGS, and in women, serum UA as well as age, body mass index, drinking status, diastolic blood pressure, and eGFRCKDEPI were independently and significantly associated with HGS. In women, age and multivariate-adjusted HGS were significantly higher in the Quartile-3 (4.8-5.4 mg/dL) and Quartile-4 groups (5.5-9.3 mg/dL) of serum UA than in the lower groups (0.7-4.7 mg/dL). These results suggest that serum UA may have a protective role in aging-associated decline in muscle strength in community-dwelling elderly women. PMID:27078883

  11. Serum Uric Acid Is Positively Associated with Handgrip Strength among Japanese Community-Dwelling Elderly Women

    PubMed Central

    Kawamoto, Ryuichi; Ninomiya, Daisuke; Kasai, Yoshihisa; Kusunoki, Tomo; Ohtsuka, Nobuyuki; Kumagi, Teru; Abe, Masanori

    2016-01-01

    Serum uric acid (UA) has strong anti-oxidant properties. Muscle strength and mass decrease with age, and recently, this decrease has been defined as sarcopenia. Sarcopenia may be triggered by oxidative stress. We investigated whether serum UA is associated with handgrip strength (HGS), which is a useful indicator of sarcopenia, among Japanese community-dwelling elderly persons. The present study included 602 men aged 72 ± 7 years and 847 women aged 71 ± 6 years from a rural village. We examined the cross-sectional relationship between serum UA and HGS. In both genders, HGS increased significantly with increased serum UA levels. A multiple linear regression analysis using HGS as an objective variable and various confounding factors as explanatory variables showed that in men age, drinking status, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and estimated glomerular filtration ratio (eGFRCKDEPI) were independently and significantly associated with HGS, and in women, serum UA as well as age, body mass index, drinking status, diastolic blood pressure, and eGFRCKDEPI were independently and significantly associated with HGS. In women, age and multivariate-adjusted HGS were significantly higher in the Quartile-3 (4.8–5.4 mg/dL) and Quartile-4 groups (5.5–9.3 mg/dL) of serum UA than in the lower groups (0.7–4.7 mg/dL). These results suggest that serum UA may have a protective role in aging-associated decline in muscle strength in community-dwelling elderly women. PMID:27078883

  12. Influence of Organic Acids from the Oral Biofilm on the Bond Strength of Self-Etch Adhesives to Dentin.

    PubMed

    Amaral, Cristiane Mariote; Correa, Danielly de Sá; Miragaya, Luciana Meirelles; Silva, Eduardo Moreira da

    2015-10-01

    The aim of this study was to evaluate the microtensile bond strength of self-etch adhesive systems to dentin after storage in acids from oral biofilm. Three adhesive systems were used in the study: a two-step self-etch adhesive for use with a silorane-based resin composite (Filtek P90 adhesive system - P90), a two-step self-etch adhesive (Clearfil SE Bond - CSE) and a one-step self-etch adhesive (Adper Easy One - AEO). The bond strength of these products was evaluated by bonding resin composite (Filtek Z350 for CSE and AEO; and Filtek P90 for P90) to 90 bovine dentin tooth fragments, according to the manufacturer's instructions. After 24 h of water storage at 37 °C, the specimens were sectioned into beams (1 mm2) divided and stored in distilled water, lactic acid and propionic acid, for 7 and 30 days. After storage, the specimens were tested for microtensile bond strength. Data were analyzed by three-way ANOVA and Tukey´s test (α=0.05). CSE presented the highest microtensile bond strength after storage in distilled water for 7 and 30 days. The microtensile bond strength of all adhesive systems was lower after storage in lactic acid and propionic acid than after water storage. Significant difference was not found between storage times. PMID:26647935

  13. Multi-site study of diffusion metric variability: characterizing the effects of site, vendor, field strength, and echo time using the histogram distance

    NASA Astrophysics Data System (ADS)

    Helmer, K. G.; Chou, M.-C.; Preciado, R. I.; Gimi, B.; Rollins, N. K.; Song, A.; Turner, J.; Mori, S.

    2016-03-01

    MRI-based multi-site trials now routinely include some form of diffusion-weighted imaging (DWI) in their protocol. These studies can include data originating from scanners built by different vendors, each with their own set of unique protocol restrictions, including restrictions on the number of available gradient directions, whether an externally generated list of gradient directions can be used, and restrictions on the echo time (TE). One challenge of multi-site studies is to create a common imaging protocol that will result in a reliable and accurate set of diffusion metrics. The present study describes the effect of site, scanner vendor, field strength, and TE on two common metrics: the first moment of the diffusion tensor field (mean diffusivity, MD), and the fractional anisotropy (FA). We have shown in earlier work that ROI metrics and the mean of MD and FA histograms are not sufficiently sensitive for use in site characterization. Here we use the distance between whole brain histograms of FA and MD to investigate within- and between-site effects. We concluded that the variability of DTI metrics due to site, vendor, field strength, and echo time could influence the results in multi-center trials and that histogram distance is sensitive metrics for each of these variables.

  14. Promoter strength of folic acid synthesis genes affects sulfa drug resistance in Saccharomyces cerevisiae.

    PubMed

    Iliades, Peter; Berglez, Janette; Meshnick, Steven; Macreadie, Ian

    2003-01-01

    The enzyme dihydropteroate synthase (DHPS) is an important target for sulfa drugs in both prokaryotic and eukaryotic microbes. However, the understanding of DHPS function and the action of antifolates in eukaryotes has been limited due to technical difficulties and the complexity of DHPS being a part of a bifunctional or trifunctional protein that comprises the upstream enzymes involved in folic acid synthesis (FAS). Here, yeast strains have been constructed to study the effects of FOL1 expression on growth and sulfa drug resistance. A DHPS knockout yeast strain was complemented by yeast vectors expressing the FOL1 gene under the control of promoters of different strengths. An inverse relationship was observed between the growth rate of the strains and FOL1 expression levels. The use of stronger promoters to drive FOL1 expression led to increased sulfamethoxazole resistance when para-aminobenzoic acid (pABA) levels were elevated. However, high FOL1 expression levels resulted in increased susceptibility to sulfamethoxazole in pABA free media. These data suggest that up-regulation of FOL1 expression can lead to sulfa drug resistance in Saccharomyces cerevisiae.

  15. High ionic strength narrows the population of sites participating in protein ion-exchange adsorption: A single-molecule study

    PubMed Central

    Kisley, Lydia; Chen, Jixin; Mansur, Andrea P.; Dominguez-Medina, Sergio; Kulla, Eliona; Kang, Marci; Shuang, Bo; Kourentzi, Katerina; Poongavanam, Mohan-Vivekanandan; Dhamane, Sagar; Willson, Richard C.; Landes, Christy F.

    2014-01-01

    The retention and elution of proteins in ion-exchange chromatography is routinely controlled by adjusting the mobile phase salt concentration. It has repeatedly been observed, as judged from adsorption isotherms, that the apparent heterogeneity of adsorption is lower at more-eluting, higher ionic strength. Here, we present an investigation into the mechanism of this phenomenon using a single-molecule, super-resolution imaging technique called motion-blur Points Accumulation for Imaging in Nanoscale Topography (mbPAINT). We observed that the number of functional adsorption sites was smaller at high ionic strength and that these sites had reduced desorption kinetic heterogeneity, and thus narrower predicted elution profiles, for the anion-exchange adsorption of α-lactalbumin on an agarose-supported, clustered-charge ligand stationary phase. Explanations for the narrowing of the functional population such as inter-protein interactions and protein or support structural changes were investigated through kinetic analysis, circular dichroism spectroscopy, and microscopy of agarose microbeads, respectively. The results suggest the reduction of heterogeneity is due to both electrostatic screening between the protein and ligand and tuning the steric availability within the agarose support. Overall, we have shown that single molecule spectroscopy can aid in understanding the influence of ionic strength on the population of functional adsorbent sites participating in the ion-exchange chromatographic separation of proteins. PMID:24751557

  16. From stiffness to strength in site response predictions: Formulation and validation of a hybrid hyperbolic nonlinear soil model

    NASA Astrophysics Data System (ADS)

    Asimaki, D.; Shi, J.

    2015-12-01

    Nonlinear site response analyses are becoming an increasingly important component of simulated ground motions intended for engineering applications. For regional-scale problems where geotechnical data are sparse, the challenge lies in computing nonlinear ground deformation to very strong ground shaking using a very small number of input parameters. We have developed a nonlinear model that addresses these issues: it simultaneously captures the soil's low-strain stiffness and large-strain strength of soil, and yields reliable predictions of soil response to weak and strong shaking using as sole input the shear wave velocity profile. We here present the formulation of the model and an extensive validation study based on downhole array recordings, with peak ground acceleration ranging from 0.01-0.9g. We also show that our model, referred to as hybrid hyperbolic, outperforms existing nonlinear formulations and simplified site response analyses widely used in practice for ground motions that induce more than 0.02% of soil strain (roughly equivalent to PGA higher than 0.02g). In addition to site-specific response predictions at sites with limited site characterization, the hybrid-hyperbolic model can help improve site amplification factors of ground motion prediction equations (GMPEs) by complementing the empirical data with simulated site response analyses for very strong ground shaking; as well as physics-based ground motion simulations, particularly for deeper sedimentary sites with low resonant frequencies.

  17. Influence of the ionic strength of acidic background electrolytes on the separation of proteins by capillary electrophoresis.

    PubMed

    Bekri, Samya; Leclercq, Laurent; Cottet, Hervé

    2016-02-01

    The ionic strength is one of the key parameters for optimizing CE separations. However, only a few data are available in the literature about the ionic strength effect on the separation of proteins. The effect of ionic strength on separation performances is rather complex since many different parameters are involved: such as the protein effective mobility, the electroosmotic mobility, the separation efficiency via the electromigration dispersion, as well as the viscosity and temperature of the background electrolyte. In the present work, the influence of ionic strength on the electrophoretic separation of five model proteins has been investigated in acidic conditions, on successive multi-ionic layers coated capillary, in counter-electroosmotic mode with anodic electroosmotic flow. The decrease in effective and electroosmotic mobilities with increasing ionic strength were compared using the slope-plot approach, which is very helpful for understanding the observed changes in apparent selectivity and resolution. The relative decrease of the protein effective mobility was about 30-40% of the mobility determined at 5mM ionic strength per ionic strength decade. It was found that relatively low ionic strength (∼5-10mM) was preferable to optimize the overall separation of the five model proteins. PMID:26780847

  18. Specific Initiation Site for Simian Virus 40 Deoxyribonucleic Acid Replication

    PubMed Central

    Thoren, Marilyn M.; Sebring, Edwin D.; Salzman, Norman P.

    1972-01-01

    Replicating simian virus 40 (SV40) deoxyribonucleic acid (DNA) molecules have been isolated under conditions in which the newly synthesized DNA is uniformly labeled with 3H-thymidine. These newly synthesized strands are released from the replicative intermediate molecules by alkaline treatment, and it has been possible to isolate single-stranded SV40 DNA which varies in size from 157,000 daltons (from molecules that are 10% replicated) to 1,360,000 daltons (85% replicated). The rates of duplex formation of newly synthesized DNA have been used to relate their genetic complexity to the extent of DNA replication. As DNA replication proceeds, the time required to effect 50% renaturation of the newly synthesized DNA increases at a proportional rate. The data establish that DNA replication is not initiated at random, but rather that there is a single specific initiation site for DNA replication. PMID:4342054

  19. Catalytic anisotropy of MoO/sub 3/ in oxidation reactions in the light of bond-strength model of active sites

    SciTech Connect

    Ziolkowski, J.

    1983-04-01

    Catalytic anisotropy of molybdic oxide in oxidation of propylene has been observed. High selectivity to acrolein in the indicated reaction was ascribed to the (100) plane while the (010) plane was found to yield CO/sub 2/. The critical discussion of Volta's experimental data, performed in this paper, has shown that they may be interpreted in three alternative ways, differing in the ascription of the reaction products to the crystallographic planes. Different crystallographic planes exposed by the grains of MoO/sub 3/ have been analyzed in terms of the bond-strength model of active sites developed under the following main assumptions: (i) the reaction path depends on the number and configuration of the active oxygen atoms in the vicinity of the adsorption site, and (ii) the individual catalytic activity of a given surface oxygen atom is proportional to the reciprocal sum of the strength of the bonds to it from the adjacent cations. The analysis provided the arguments to indicate the most probable reaction pattern. According to it the main products expected to be formed in oxidation of propylene on different planes of MoO/sub 3/ are (100), CO, CO/sub 2/; (001), acrolein (acrylic acid, C/sub 2/-O, CO, CO/sub 2/--mainly at longer contact time); (101) and (101), acrolein; (010), inactive (possible minor yield of hexadiene and benzene).

  20. Chemical characteristics and sources of organic acids in precipitation at a semi-urban site in Southwest China

    NASA Astrophysics Data System (ADS)

    Zhang, Y. L.; Lee, X. Q.; Cao, F.

    2011-01-01

    In order to investigate the chemical characteristics and sources of organic acids in precipitation in Southwest China, 105 rainwater samples were collected at a semi-urban site in Anshun from June 2007 to June 2008. Organic acids and major anions were analyzed along with pH and electrical conductivity. The pH values varied from 3.57 to 7.09 for all the rainfall events sampled, with an average of 4.67 which was typical acidic value. Formic, acetic and oxalic acids were found to be the predominant carboxylic acids and their volume weighted average (VWA) concentrations were 8.77, 6.93 and 2.84 μmol l -1, respectively. These organic acids were estimated to account for 8.1% to the total free acidity (TFA) in precipitation. The concentrations of the majority organic acids at studied site had a clear seasonal pattern, reaching higher levels during the non-growing season than those in growing season, which was attributed to dilution effect of heavy rainfall during the growing season. The seasonal variation of wet deposition flux of these organic acids confirmed higher source strength of biogenic emissions from vegetation during the growing season. Formic-to-acetic acids ratio (F/A), an indicator of primary versus secondary sources of these organic acids, suggested that primary sources from vehicular emission, biomass burning, soil and vegetation emissions were dominant sources. In addition, the lowest concentrations of organic acids were found under type S, when air masses originated from the marine (South China Sea) during Southern Asian Monsoon period. And the highest concentrations were observed in precipitation events from Northeast China (type NE), prevailing mostly during winter with the lowest rainfall.

  1. EFFECTS OF PH, SOLID/SOLUTION RATIO, IONIC STRENGTH, AND ORGANIC ACIDS ON PB AND CD SOPRTION ON KAOLINITE

    EPA Science Inventory

    Potentiometric and ion-selective electrode titrations together with batch sorption/desorption experiments, were performed to explain the aqueous and surface complexation reactions between kaolinite, Pb, Cd and three organic acids. Variables included pH, ionic strength, metal conc...

  2. Ionic Strength Effect on the Rate of Reduction of Hexacyanoferrate (III) by Ascorbic Acid: A Physical Chemistry Laboratory Experiment.

    ERIC Educational Resources Information Center

    Watkins, Kenneth W.; Olson, June A.

    1980-01-01

    Describes a physical chemistry experiment that allows students to test the effect of ionic strength on the rates of a reaction between ions. The reduction of hexacyanoferrate III by ascorbic acid is detailed. Comparisons with the iodine clock reaction are made. (CS)

  3. Direct inference of site strength in basic solids upon CO2 adsorption: enthalpy-entropy compensation effects.

    PubMed

    Pera-Titus, M

    2016-08-10

    The adsorption of CO2 coupled to calorimetry is a state-of-the-art technique for characterizing the basic properties of solids. In this paper, we show that the differential heat and entropy curves measured upon CO2 adsorption on a basic solid can be reasonably estimated from a single CO2 isotherm with no need for any independent heat (calorimetric) measurement. Our method relies on two important observations: (1) formulation of generalized F-H-TS thermodynamic isotherms, the former (F) being directly generated from the raw CO2 isotherms, and (2) the presence of unexpected enthalpy-entropy compensation effects upon CO2 adsorption linking the integral enthalpy and entropy of adsorption until saturation for different solids. Our thermodynamic method has been validated using a broad library of basic solids with variable site strength and heterogeneity. Finally, a new scale of basicity is proposed using the parameters fitted from the thermodynamic isotherm (free energy basis) as descriptors of basic strength. This method opens an avenue to the inference of site strength of basic solids without the need for expensive calorimeters.

  4. Direct inference of site strength in basic solids upon CO2 adsorption: enthalpy-entropy compensation effects.

    PubMed

    Pera-Titus, M

    2016-08-10

    The adsorption of CO2 coupled to calorimetry is a state-of-the-art technique for characterizing the basic properties of solids. In this paper, we show that the differential heat and entropy curves measured upon CO2 adsorption on a basic solid can be reasonably estimated from a single CO2 isotherm with no need for any independent heat (calorimetric) measurement. Our method relies on two important observations: (1) formulation of generalized F-H-TS thermodynamic isotherms, the former (F) being directly generated from the raw CO2 isotherms, and (2) the presence of unexpected enthalpy-entropy compensation effects upon CO2 adsorption linking the integral enthalpy and entropy of adsorption until saturation for different solids. Our thermodynamic method has been validated using a broad library of basic solids with variable site strength and heterogeneity. Finally, a new scale of basicity is proposed using the parameters fitted from the thermodynamic isotherm (free energy basis) as descriptors of basic strength. This method opens an avenue to the inference of site strength of basic solids without the need for expensive calorimeters. PMID:27468818

  5. Effect of Peracetic Acid as A Final Rinse on Push Out Bond Strength of Root Canal Sealers to Root Dentin

    PubMed Central

    Gaddala, Naresh; Veeramachineni, Chandrasekhar

    2015-01-01

    Background Smear layer which was formed during the instrumentation of root canals hinders the penetration of root canal sealers to root dentin and affect the bond strength of root canal sealers to root dentin. Final irrigant such as demineralizing agents are used to remove the inorganic portion of the smear layer. In the present study, peracetic acid used as a final rinse, to effect the bond strength of root canal sealers to root dentin. Aim The purpose of the present study was to evaluate the efficacy of peracetic acid as a final irrigant on bond strength of root canal sealers to root dentin. Materials and Methods Sixty six freshly extracted human single rooted mandibular premolars were used for this study. After decoronation the samples were instrumented with Protaper upto F3 and irrigated with 5.25% NaOcl. The teeth were then divided into three groups based on final irrigant used: Group-1(control group) Canals were irrigated with distilled water. Group-2: Canals were irrigated with peracetic acid. Group-3: Canals were irrigated with smear clear. Each group was further divided into three subgroups (n=30) based on the sealer used to obturate the canals. Subgroup-1: kerr, Subgroup-2: Apexit plus, Subgroup-3: AH PLUS. Each sealer was mixed and coated to master cone and placed in the canal. The bonding between sealer and dentin surface was evaluated using push out bond strength by universal testing machine. The mean bond strength values of each group were statistically evaluated using Two-way ANOVA followed by Tukey post-hoc test. Results Significant difference was found among the bond strength of the sealers. But, there is no statistically significant difference between the groups irrigated with peracetic acid and smear clear compared to control group. AH Plus showed highest bond strength irrespective of the final irrigant used. Conclusion Peracetic acid when employed as final irrigant improved the bond strength of root canal sealers compared to control group but

  6. [Study on porcelain veneer restorations. 2. Influence of hydrofluoric acid on bonding strength at the porcelain-resin interface].

    PubMed

    Gomi, A; Ikeda, M; Takeuchi, N; Ban, Y; Kamiya, K; Kanamori, K; Asai, T; Senda, A

    1990-06-01

    Recently, porcelain veneer restoratives have been introduced to the general practice, and their clinical performances have been confirmed through many longterm clinical investigations. It is expected that porcelain veneer restorations will perform successfully in esthetic, conservative and abhesive dentistry. It is an well known fact that the micro-mechanical bonding strength at the porcelain-resin interface which is achieved through the application of hydrofluoric acid to the porcelain surface is quite a strong bonding mechanism. However, there are very few studies reporting on the acid treatment of porcelain surfaces. The authors have been studying the influence of hydrofluoric acid on porcelain surfaces, and in our previous report we reported, the degrees of corroded porcelain treated with different concentrations of hydrofluoric acid for different durations of application. In the present study, shear bonding strength was measured between resin cements and porcelain surfaces treated with different concentrations (4, 6, 8%) of hydrofluoric acid and for different durations (1 to 24 min.), and the appropriate treatment of porcelain surfaces with regard to the bonding strength was determined. The results obtained were as follows. 1. As the treating time increased with any concentration (4, 6, 8%) of hydrofluoric acid, corrosion of the porcelain surface became more intense. Hardly any evidence of corrosion was observed on any porcelain surface treated for one minute, so it seems that the treatment of porcelain surfaces using 4 to 8% hydrofluoric acids should be continued for over three minutes. 2. Observation of the surface profile by SEM showed no significant differences between the surfaces treated for 3, 6, 12 and 24 minutes. 3. It was not clear as to how the differences of hydrofluoric acid concentrations (4, 6, 8%) plus the differences in the kinds of porcelain (Super Porcelain AAA, NORITAKE Co. Ltd., VMK 68, Vita Zahnfabrik Gmbh & Co., Cosmotech Porcelain, G

  7. High Folic Acid Intake during Pregnancy Lowers Body Weight and Reduces Femoral Area and Strength in Female Rat Offspring

    PubMed Central

    Huot, Pedro S. P.; Dodington, David W.; Mollard, Rebecca C.; Reza-López, Sandra A.; Sánchez-Hernández, Diana; Cho, Clara E.; Kuk, Justin; Ward, Wendy E.; Anderson, G. Harvey

    2013-01-01

    Rats fed gestational diets high in multivitamin or folate produce offspring of altered phenotypes. We hypothesized that female rat offspring born to dams fed a gestational diet high in folic acid (HFol) have compromised bone health and that feeding the offspring the same HFol diet attenuates these effects. Pregnant rats were fed diets with either recommended folic acid (RFol) or 10-fold higher folic acid (HFol) amounts. Female offspring were weaned to either the RFol or HFol diet for 17 weeks. HFol maternal diet resulted in lower offspring body weights (6%, P = 0.03) and, after adjusting for body weight and femoral length, smaller femoral area (2%, P = 0.03), compared to control diet. After adjustments, HFol pup diet resulted in lower mineral content (7%, P = 0.01) and density (4%, P = 0.002) of lumbar vertebra 4 without differences in strength. An interaction between folate content of the dam and pup diets revealed that a mismatch resulted in lower femoral peak load strength (P = 0.01) and stiffness (P = 0.002). However, the match in folate content failed to prevent lower weight gain. In conclusion, HFol diets fed to rat dams and their offspring affect area and strength of femurs and mineral quantity but not strength of lumbar vertebrae in the offspring. PMID:23781391

  8. Effect of acid etching on bond strength of nanoionomer as an orthodontic bonding adhesive

    PubMed Central

    Khan, Saba; Verma, Sanjeev K.; Maheshwari, Sandhya

    2015-01-01

    Aims: A new Resin Modified Glass Ionomer Cement known as nanoionomer containing nanofillers of fluoroaluminosilicate glass and nanofiller 'clusters' has been introduced. An in-vitro study aimed at evaluating shear bond strength (SBS) and adhesive remnant index (ARI) of nanoionomer under etching/unetched condition for use as an orthodontic bonding agent. Material and Methods: A total of 75 extracted premolars were used, which were divided into three equal groups of 25 each: 1-Conventional adhesive (Enlight Light Cure, SDS, Ormco, CA, USA) was used after and etching with 37% phosphoric acid for 30 s, followed by Ortho Solo application 2-nanoionomer (Ketac™ N100, 3M, ESPE, St. Paul, MN, USA) was used after etching with 37% phosphoric acid for 30 s 3-nanoionomer was used without etching. The SBS testing was performed using a digital universal testing machine (UTM-G-410B, Shanta Engineering). Evaluation of ARI was done using scanning electron microscopy. The SBS were compared using ANOVA with post-hoc Tukey test for intergroup comparisons and ARI scores were compared with Chi-square test. Results: ANOVA (SBS, F = 104.75) and Chi-square (ARI, Chi-square = 30.71) tests revealed significant differences between groups (P < 0.01). The mean (SD) SBS achieved with conventional light cure adhesive was significantly higher (P < 0.05) (10.59 ± 2.03 Mpa, 95% CI, 9.74-11.41) than the nanoionomer groups (unetched 4.13 ± 0.88 Mpa, 95% CI, 3.79-4.47 and etched 9.32 ± 1.87 Mpa, 95% CI, 8.58-10.06). However, nanoionomer with etching, registered SBS in the clinically acceptable range of 5.9–7.8 MPa, as suggested by Reynolds (1975). The nanoionomer groups gave significantly lower ARI values than the conventional adhesive group. Conclusion: Based on this in-vitro study, nanoionomer with etching can be successfully used as an orthodontic bonding agent leaving less adhesive remnant on enamel surface, making cleaning easier. However, in-vivo studies are needed to confirm the validity

  9. A Sialic Acid Binding Site in a Human Picornavirus

    PubMed Central

    Frank, Martin; Hähnlein-Schick, Irmgard; Ekström, Jens-Ola; Arnberg, Niklas; Stehle, Thilo

    2014-01-01

    The picornaviruses coxsackievirus A24 variant (CVA24v) and enterovirus 70 (EV70) cause continued outbreaks and pandemics of acute hemorrhagic conjunctivitis (AHC), a highly contagious eye disease against which neither vaccines nor antiviral drugs are currently available. Moreover, these viruses can cause symptoms in the cornea, upper respiratory tract, and neurological impairments such as acute flaccid paralysis. EV70 and CVA24v are both known to use 5-N-acetylneuraminic acid (Neu5Ac) for cell attachment, thus providing a putative link between the glycan receptor specificity and cell tropism and disease. We report the structures of an intact human picornavirus in complex with a range of glycans terminating in Neu5Ac. We determined the structure of the CVA24v to 1.40 Å resolution, screened different glycans bearing Neu5Ac for CVA24v binding, and structurally characterized interactions with candidate glycan receptors. Biochemical studies verified the relevance of the binding site and demonstrated a preference of CVA24v for α2,6-linked glycans. This preference can be rationalized by molecular dynamics simulations that show that α2,6-linked glycans can establish more contacts with the viral capsid. Our results form an excellent platform for the design of antiviral compounds to prevent AHC. PMID:25329320

  10. Strength distribution of fatigue crack initiation sites in an Al-Li alloy

    NASA Astrophysics Data System (ADS)

    Zhai, T.

    2006-10-01

    The stress-number of cycles to failure (S-N) curves were measured along the short-transverse (S) and rolling (L) directions of a hot-cross-rolled AA 8090 Al-Li alloy plate (45-mm thick). The alloy was solution heat treated, quenched in water, strained by 6 pct, and peak aged. Fatigue tests were carried out in four-point bend at room temperature, 20 Hz, R=0.1, in air. It was found that the fatigue limits in the S and L directions were 147 and 197 MPa, respectively. The crack population on the surface of a sample at failure increased with the applied stress level and was found to be a Weibull function of the applied maximum stress in this alloy. The strength distribution of fatigue weakest links, where cracks were initiated, was derived from the Weibull function determined by the experimental data. The fatigue weakest-link density was defined as the crack population per unit area at a stress level close to the ultimate tensile stress and can be regarded as a materials property. The density and strength distribution of fatigue weakest links were found to be markedly different between the L and S directions, accounting for the difference in fatigue limit between the directions in this alloy. They were also found to be different between S-L and S-T samples, and between L-T and L-S samples of this alloy, which could not be revealed by the corresponding S-N curves measured. These differences were due to the anisotropy of the microstructures in different directions in this alloy.

  11. Effects of ultraviolet irradiation on bonding strength between Co-Cr alloy and citric acid-crosslinked gelatin matrix.

    PubMed

    Inoue, Motoki; Sasaki, Makoto; Katada, Yasuyuki; Taguchi, Tetsushi

    2014-02-01

    Novel techniques for creating a strong bond between polymeric matrices and biometals are required. We immobilized polymeric matrices on the surface of biometal for drug-eluting stents through covalent bond. We performed to improve the bonding strength between a cobalt-chromium alloy and a citric acid-crosslinked gelatin matrix by ultraviolet irradiation on the surface of cobalt-chromium alloy. The ultraviolet irradiation effectively generated hydroxyl groups on the surface of the alloy. The bonding strength between the gelatin matrix and the alloy before ultraviolet irradiation was 0.38 ± 0.02 MPa, whereas it increased to 0.48 ± 0.02 MPa after ultraviolet irradiation. Surface analysis showed that the citric acid derivatives occurred on the surface of the cobalt-chromium alloy through ester bond. Therefore, ester bond formation between the citric acid derivatives active esters and the hydroxyl groups on the cobalt-chromium alloy contributed to the enhanced bonding strength. Ultraviolet irradiation and subsequent immobilization of a gelatin matrix using citric acid derivatives is thus an effective way to functionalize biometal surfaces.

  12. Sensitive methods for estimating the anchoring strength of nematic liquid crystals on Langmuir-Blodgett monolayers of fatty acids

    SciTech Connect

    Fazio, Valentina S. U.; Nannelli, Francesca; Komitov, Lachezar

    2001-06-01

    The anchoring of the nematic liquid crystal N-(p-methoxybenzylidene)-p-butylaniline (MBBA) on Langmuir-Blodgett monolayers of fatty acids (COOHC{sub n}H{sub 2n+1}) was studied as a function of the length of the fatty acid alkyl chain n (n=15,17,19,21). The monolayers were deposited onto glass plates coated with indium tin oxide, which were used to assemble sandwich cells of various thicknesses that were filled with MBBA in the nematic phase. The mechanism of relaxation from the flow-induced quasiplanar to the surface-induced homeotropic alignment was studied for the four aligning monolayers. It was found that the speed of the relaxation decreases linearly with increasing length of the alkyl chain n, which suggests that the Langmuir-Blodgett film plays a role in the phenomenon. This fact was confirmed by a sensitive estimation of the anchoring strength of MBBA on the fatty acid monolayers after anchoring breaking, which takes place at the transition between two electric-field-induced turbulent states, denoted as DSM1 and DSM2 (where DSM indicates dynamic scattering mode). It was found that the threshold electric field for the anchoring breaking, which can be considered as a measure of the anchoring strength, also decreases linearly as n increases. Both methods thus possess a high sensitivity in resolving small differences in anchoring strength. In cells coated with mixed Langmuir-Blodgett monolayers of two fatty acids (n=15 and n=17) a maximum of the relaxation speed was observed when the two acids were present in equal amounts. This observation suggests an efficient method for controlling the anchoring strength in homeotropic cells by changing the ratio between the components of the surfactant film.

  13. A multi-site, randomized study of strengths-based case management with substance-abusing parolees.

    PubMed

    Prendergast, Michael; Frisman, Linda; Sacks, Joann Y; Staton-Tindall, Michele; Greenwell, Lisa; Lin, Hsiu-Ju; Cartier, Jerry

    2011-09-01

    OBJECTIVES: To test whether strengths-based case management provided during an inmate's transition from incarceration to the community increases participation in community substance abuse treatment, enhances access to needed social services, and improves drug use, crime, and HIV risk outcomes. METHODS: In a multi-site trial, inmates (men and women) in four states (n = 812) were randomly assigned (within site) to receive either Transitional Case Management (TCM group), based on strengths-based principles, or standard parole services (SR group). Data were collected at baseline and at 3 and 9 months following release from prison. Analyses compared the two groups with respect to services received and to drug use, crime, and HIV risk behavior outcomes. RESULTS: There were no significant differences between parolees in the TCM group and the SR group on outcomes related to participation in drug abuse treatment, receipt of social services, or drug use, crime, and HIV risk behaviors. For specific services (e.g., residential treatment, mental health), although significant differences were found for length of participation or for number of visits, the number of participants in these services was small and the direction of effect was not consistent. CONCLUSION: In contrast to positive findings in earlier studies of strengths-based case management with mental-health and drug-abuse clients, this study found that case management did not improve treatment participation or behavioral outcomes for parolees with drug problems. The discussion includes possible reasons for the findings and suggestions for modifications to the intervention that could be addressed in future research.

  14. Highly selective Lewis acid sites in desilicated MFI zeolites for dihydroxyacetone isomerization to lactic acid.

    PubMed

    Dapsens, Pierre Y; Mondelli, Cecilia; Pérez-Ramírez, Javier

    2013-05-01

    Desilication of commercial MFI-type (ZSM-5) zeolites in solutions of alkali metal hydroxides is demonstrated to generate highly selective heterogeneous catalysts for the aqueous-phase isomerization of biobased dihydroxyacetone (DHA) to lactic acid (LA). The best hierarchical ZSM-5 sample attains a LA selectivity exceeding 90 %, which is comparable to that of the state-of-the-art catalyst (i.e., the Sn-beta zeolite); this optimized hierarchical catalyst is recyclable over three runs. The Lewis acid sites, which are created through desilication along with the introduction of mesoporosity, are shown to play a crucial role in the formation of the desired product; these cannot be achieved by using other post-synthetic methods, such as steaming or impregnation of aluminum species. Desilication of other metallosilicates, such as Ga-MFI, also leads to high LA selectivity. In the presence of a soluble aluminum source, such as aluminum nitrate, alkaline-assisted alumination can introduce these unique Lewis acid centers in all-silica MFI zeolites. These findings highlight the potential of zeolites in the field of biomass-to-chemical conversion, and expand the applicability of desilication for the generation of selective catalytic centers. PMID:23554234

  15. Endolysosomes Are the Principal Intracellular Sites of Acid Hydrolase Activity.

    PubMed

    Bright, Nicholas A; Davis, Luther J; Luzio, J Paul

    2016-09-12

    The endocytic delivery of macromolecules from the mammalian cell surface for degradation by lysosomal acid hydrolases requires traffic through early endosomes to late endosomes followed by transient (kissing) or complete fusions between late endosomes and lysosomes. Transient or complete fusion results in the formation of endolysosomes, which are hybrid organelles from which lysosomes are re-formed. We have used synthetic membrane-permeable cathepsin substrates, which liberate fluorescent reporters upon proteolytic cleavage, as well as acid phosphatase cytochemistry to identify which endocytic compartments are acid hydrolase active. We found that endolysosomes are the principal organelles in which acid hydrolase substrates are cleaved. Endolysosomes also accumulated acidotropic probes and could be distinguished from terminal storage lysosomes, which were acid hydrolase inactive and did not accumulate acidotropic probes. Using live-cell microscopy, we have demonstrated that fusion events, which form endolysosomes, precede the onset of acid hydrolase activity. By means of sucrose and invertase uptake experiments, we have also shown that acid-hydrolase-active endolysosomes and acid-hydrolase-inactive, terminal storage lysosomes exist in dynamic equilibrium. We conclude that the terminal endocytic compartment is composed of acid-hydrolase-active, acidic endolysosomes and acid hydrolase-inactive, non-acidic, terminal storage lysosomes, which are linked and function in a lysosome regeneration cycle. PMID:27498570

  16. Effect of acid etching time and technique on bond strength of an etch-and-rinse adhesive.

    PubMed

    Faria-e-Silva, André L; Silva, João L; Almeida, Thauanna G; Veloso, Francielle B; Ribeiro, Sandra M; Andrade, Tiago D; Vilas-Boas, Bruna V; Martins, Marisa C; Menezes, Murilo S

    2011-01-01

    The aim of this study was to evaluate the effect of acid etching time and technique on bond strength of a two-step etch-and-rinse adhesive system to dentin and enamel. Thirty human third molars were mesio-distally sectioned, parallel to the long axis of each tooth, in two halves. Buccal/lingual surfaces were abraded to obtain both flat exposed enamel and dentine. The etchant was applied with and without the use of dispensing tips provided by manufacturer. When the tip was not used, the etchant was agitated (active) over the substrate or left undisturbed (passive). The etchings were done for 15 or 30s. After rinsing the acid, the adhesive XP Bond (Dentsply Caulk, Milford, DE, USA) was applied and light-cured. Resin composite cylinders were built up on dentin and enamel substrates. A shear load was applied to the samples at a crosshead speed of 0.5 mm/min until failure. Data were statistically analyzed by three-way ANOVA and Tukey test (alpha = 0.05). There was no difference between the etching techniques in bonding to enamel. Application with the tip or active without the tip promoted higher bond strength to dentin than passive application. Extending the etching time reduced the bond strength to dentin and did not alter the values for enamel. The passive application without tips produced the lowest bond strength when the etchant was applied for 15s. All techniques demonstrated similar values for application during 30s. The acid etching time and technique significantly influence the bond strength of etch-and-rinse adhesive to dentin. PMID:22010410

  17. Comparative Evaluation of Shear Bond Strength of Luting Cements to Different Core Buildup Materials in Lactic Acid Buffer Solution

    PubMed Central

    Patil, Siddharam M.; Desai, Raviraj G.; Arabbi, Kashinath C.; Prakash, Ved

    2015-01-01

    Aim and Objectives The core buildup material is used to restore badly broken down tooth to provide better retention for fixed restorations. The shear bond strength of a luting agent to core buildup is one of the crucial factors in the success of the cast restoration. The aim of this invitro study was to evaluate and compare the shear bond strength of luting cements with different core buildup materials in lactic acid buffer solution. Materials and Methods Two luting cements {Traditional Glass Ionomer luting cement (GIC) and Resin Modified Glass Ionomer luting cement (RMGIC)} and five core buildup materials {Silver Amalgam, Glass ionomer (GI), Glass Ionomer Silver Reinforced (GI Silver reinforced), Composite Resin and Resin Modified Glass Ionomer(RMGIC)} were selected for this study. Total 100 specimens were prepared with 20 specimens for each core buildup material using a stainless steel split metal die. Out of these 20 specimens, 10 specimens were bonded with each luting cement. All the bonded specimens were stored at 370c in a 0.01M lactic acid buffer solution at a pH of 4 for 7days. Shear bond strength was determined using a Universal Testing Machine at a cross head speed of 0.5mm/min. The peak load at fracture was recorded and shear bond strength was calculated. The data was statistically analysed using Two-way ANOVA followed by HOLM-SIDAK method for pair wise comparison at significance level of p<0.05. Results Two-Way ANOVA showed significant differences in bond strength of the luting cements (p<0.05) and core materials (p<0.05) and the interactions (p<0.05). Pairwise comparison of luting cements by HOLM-SIDAK test, showed that the RMGIC luting cement had higher shear bond strength values than Traditional GIC luting cement for all the core buildup materials. RMGIC core material showed higher bond strength values followed by Composite resin, GI silver reinforced, GI and silver amalgam core materials for both the luting agents. Conclusion Shear bond strength of

  18. Design of a Brønsted acid with two different acidic sites: synthesis and application of aryl phosphinic acid-phosphoric acid as a Brønsted acid catalyst.

    PubMed

    Momiyama, N; Narumi, T; Terada, M

    2015-12-11

    A Brønsted acid with two different acidic sites, aryl phosphinic acid-phosphoric acid, has been synthesized. Its catalytic performance was assessed in the hetero-Diels-Alder reaction of aldehyde hydrates with Danishefsky's diene, achieving high reaction efficiency. PMID:26445921

  19. Biodegradable poly-lactic acid based-composite reinforced unidirectionally with high-strength magnesium alloy wires.

    PubMed

    Li, X; Chu, C L; Liu, L; Liu, X K; Bai, J; Guo, C; Xue, F; Lin, P H; Chu, Paul K

    2015-05-01

    Biodegradable poly-lactic acid (PLA)--based composites reinforced unidirectionally with high-strength magnesium alloy wires (MAWs) are fabricated by a heat-compressing process and the mechanical properties and degradation behavior are studied experimentally and theoretically. The composites possess improved strengthening and toughening properties. The bending strength and impact strength of the composites with 40 vol% MAWs are 190 MPa and 150 kJ/m(2), respectively, although PLA has a low viscosity and an average molecular weight of 60,000 g/mol. The mechanical properties of the composites can be further improved by internal structure modification and interface strengthening and a numerical model incorporating the equivalent section method (ESM) is proposed for the bending strength. Micro arc oxidization (MAO) of the MAWs is an effective interfacial strengthening method. The composites exhibit high strength retention during degradation and the PLA in the composite shows a smaller degradation rate than pure PLA. The novel biodegradable composites have large potential in bone fracture fixation under load-bearing conditions. PMID:25725562

  20. How to improve the on-site MOE assessment of old timber beams combining NDT and visual strength grading

    NASA Astrophysics Data System (ADS)

    Cavalli, Alberto; Togni, Marco

    2013-09-01

    For the conservation and restoration of old timber structures, the knowledge of the mechanical properties of each element is fundamental. For this reason, various nondestructive techniques were developed and investigated since the 1990s. Some of them provide very good results, but the solutions and the proposed models were applied only in few circumstances as a consequence of the on-site restrictive working conditions: no possibility to remove the elements, limitation to ends and faces accessibility and unknown density. The on-site inspection, including the visual strength grading, has been identified as the first step for the timber assessment. In this research, 13 old timber members of Silver Fir (Abies alba Mill.) were visually graded and tested with different nondestructive techniques for the density and modulus of elasticity (MOE) estimation: flexural and longitudinal vibrational tests, stress wave transmission time and Pilodyn penetration depth. The timber elements were also tested in a four-point bending test to determinate the local and global MOE. Finally, a reliable method, applicable to the limiting on-site conditions, was proposed and the results were showed.

  1. Effect of hydrofluoric acid etching duration on the roughness and flexural strength of a lithium disilicate-based glass ceramic.

    PubMed

    Zogheib, Lucas Villaça; Bona, Alvaro Della; Kimpara, Estevão Tomomitsu; McCabe, John F

    2011-01-01

    The aim of this study was to examine the effect of different acid etching times on the surface roughness and flexural strength of a lithium disilicate-based glass ceramic. Ceramic bar-shaped specimens (16 mm x 2 mm x 2 mm) were produced from ceramic blocks. All specimens were polished and sonically cleaned in distilled water. Specimens were randomly divided into 5 groups (n=15). Group A (control) no treatment. Groups B-E were etched with 4.9% hydrofluoric acid (HF) for 4 different etching periods: 20 s, 60 s, 90 s and 180 s, respectively. Etched surfaces were observed under scanning electron microscopy. Surface profilometry was used to examine the roughness of the etched ceramic surfaces, and the specimens were loaded to failure using a 3-point bending test to determine the flexural strength. Data were analyzed using one-way ANOVA and Tukey's test (?=0.05). All etching periods produced significantly rougher surfaces than the control group (p<0.05). Roughness values increased with the increase of the etching time. The mean flexural strength values were (MPa): A=417 ± 55; B=367 ± 68; C=363 ± 84; D=329 ± 70; and E=314 ± 62. HF etching significantly reduced the mean flexural strength as the etching time increased (p=0.003). In conclusion, the findings of this study showed that the increase of HF etching time affected the surface roughness and the flexural strength of a lithium disilicate-based glass ceramic, confirming the study hypothesis.

  2. Site specific incorporation of keto amino acids into proteins

    DOEpatents

    Schultz, Peter G.; Wang, Lei

    2009-04-28

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate keto amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with keto amino acids using these orthogonal pairs.

  3. Site specific incorporation of keto amino acids into proteins

    DOEpatents

    Schultz, Peter G.; Wang, Lei

    2008-10-07

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate keto amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with keto amino acids using these orthogonal pairs.

  4. Site specific incorporation of keto amino acids into proteins

    SciTech Connect

    Schultz, Peter G.; Wang, Lei

    2012-02-14

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate keto amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with keto amino acids using these orthogonal pairs.

  5. Site specific incorporation of keto amino acids into proteins

    SciTech Connect

    Schultz, Peter G.; Wang, Lei

    2011-12-06

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate keto amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with keto amino acids using these orthogonal pairs.

  6. Site specific incorporation of keto amino acids into proteins

    SciTech Connect

    Schultz, Peter G.; Wang, Lei

    2011-03-22

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate keto amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with keto amino acids using these orthogonal pairs.

  7. Humic Acid Effects on the Transport of Colloidal Particles in Unsaturated Porous Media: Humic Acid Dosage, pH, and Ionic Strength Dependence

    NASA Astrophysics Data System (ADS)

    Morales, V. L.; Gao, B.; Steenhuis, T. S.

    2008-12-01

    Soil colloids and biocolloids can facilitate contaminant transport within the soil profile through the complexation of pollutants previously thought to have limited mobility. Dissolved organic substances are qualitatively known to alter the behavior of colloids and surface chemistry of soil particles in aquatic environments when adsorbed to their surfaces. Specifically, it has been observed that even small amounts of adsorbed humic acids result in a pronounced increase in colloid mobility in saturated porous systems, presumably by a combination of electrostatic and steric stabilization. However, the degree to which adsorbed humic acids stabilize colloidal suspension is highly sensitive to the system's solution chemistry; mainly in terms of pH, ionic strength, and metal ions present. The objective of this study is to expound quantitatively on the role that combined stabilizing and destabilizing solution chemistry components have on humic acid-colloid transport in unsaturated media by isolating experimentally some underlying mechanisms that regulate colloid transport in realistic aquatic systems. We hypothesize that in chemically heterogeneous porous media, with ionic strength values above 0 and pH ranges from 4 to 9, the effect of humic acid on colloid suspensions cannot be simply characterized by increased stability and mobility. That a critical salt concentration must exists for a given humic acid concentration and pH, above which the network of humic acid collapses by forming coordination complexes with other suspended or adsorbed humic acids, thus increasing greatly the retention of colloids in the porous medium by sweep flocculation. In addition, capillary forces in unsaturated media may contribute further to overcome repulsive forces that prevent flocculation of humic acid-colloid complexes. The experimental work in this study will include: jar tests to determine critical solution concentration combinations for desired coagulation/flocculation rates, column

  8. Shear bond strength of orthodontic brackets after acid-etched and erbium-doped yttrium aluminum garnet laser-etched

    PubMed Central

    Alavi, Shiva; Birang, Reza; Hajizadeh, Fatemeh

    2014-01-01

    Background: Laser ablation has been suggested as an alternative method to acid etching; however, previous studies have obtained contrasting results. The purpose of this study was to compare the shear bond strength (SBS) and fracture mode of orthodontic brackets that are bonded to enamel etched with acid and erbium-doped yttrium aluminum garnet (Er:YAG) laser. Materials and Methods: In this experimental in vitro study, buccal surfaces of 15 non-carious human premolars were divided into mesial and distal regions. Randomly, one of the regions was etched with 37% phosphoric acid for 15 s and another region irradiated with Er:YAG laser at 100 mJ energy and 20 Hz frequency for 20 s. Stainless steel brackets were then bonded using Transbond XT, following which all the samples were stored in distilled water for 24 h and then subjected to 500 thermal cycles. SBS was tested by a chisel edge, mounted on the crosshead of universal testing machine. After debonding, the teeth were examined under ×10 magnification and adhesive remnant index (ARI) score determined. SBS and ARI scores of the two groups were then compared using t-test and Mann-Whitney U test. Significant level was set at P < 0.05. Results: The mean SBS of the laser group (16.61 ± 7.7 MPa) was not significantly different from that of the acid-etched group (18.86 ± 6.09 MPa) (P = 0.41). There was no significant difference in the ARI scores between two groups (P = 0.08). However, in the laser group, more adhesive remained on the brackets, which is not suitable for orthodontic purposes. Conclusion: Laser etching at 100 mJ energy produced bond strength similar to acid etching. Therefore, Er:YAG laser may be an alternative method for conventional acid-etching. PMID:25097641

  9. Modeling and predicting the tensile strength of poly (lactic acid)/graphene nanocomposites by using support vector regression

    NASA Astrophysics Data System (ADS)

    Cheng, W. D.; Cai, C. Z.; Luo, Y.; Li, Y. H.; Zhao, C. J.

    2016-04-01

    According to an experimental dataset under different process parameters, support vector regression (SVR) combined with particle swarm optimization (PSO) for its parameter optimization was employed to establish a mathematical model for prediction of the tensile strength of poly (lactic acid) (PLA)/graphene nanocomposites. Four variables, while graphene loading, temperature, time and speed, were employed as input variables, while tensile strength acted as output variable. Using leave-one-out cross validation test of 30 samples, the maximum absolute percentage error does not exceed 1.5%, the mean absolute percentage error (MAPE) is only 0.295% and the correlation coefficient (R2) is as high as 0.99. Compared with the results of response surface methodology (RSM) model, it is shown that the estimated errors by SVR are smaller than those achieved by RSM. It revealed that the generalization ability of SVR is superior to that of RSM model. Meanwhile, multifactor analysis is adopted for investigation on significances of each experimental factor and their influences on the tensile strength of PLA/graphene nanocomposites. This study suggests that the SVR model can provide important theoretical and practical guide to design the experiment, and control the intensity of the tensile strength of PLA/graphene nanocomposites via rational process parameters.

  10. Effect of salicylhydroxamic acid on endosperm strength and embryo growth of Lactuca sativa L. cv Waldmann's Green seeds

    NASA Technical Reports Server (NTRS)

    Brooks, C. A.; Mitchell, C. A.

    1988-01-01

    Salicylhydroxamic acid (SHAM) stimulated germination of photosensitive lettuce (Lactuca sativa L. cv Waldmann's Green) seeds in darkness. To determine whether SHAM acts on the embryo or the endosperm, we investigated separately effects of SHAM on growth potential of isolated embryos as well as on endosperm strength. Embryo growth potential was quantified by incubating decoated embryos in various concentrations of osmoticum and measuring subsequent radicle elongation. Growth potential of embryos isolated from seeds pretreated with 4 millimolar SHAM was equal to that of untreated controls. Rupture strength of endosperm tissue excised from seeds pretreated with SHAM was 33% less than that of controls in the micropylar region. To determine if the embryo must be in contact with the endosperm of SHAM to weaken the endosperm, some endosperms were incubated with SHAM only after dissection from seeds. Rupture strength of SHAM-treated, isolated endosperms in the micropylar region was 25% less than that of untreated controls. There was no difference in rupture strength in the cotyledonary region of endosperm isolated from seeds treated with SHAM in buffer or buffer alone. SHAM therefore stimulates germination not by enhancing embryo growth potential, but by weakening the micropylar region of the endosperm enclosing the embryo.

  11. Effect of Salicylhydroxamic Acid on Endosperm Strength and Embryo Growth of Lactuca sativa L. cv Waldmann's Green Seeds 1

    PubMed Central

    Brooks, Carolyn Anne; Mitchell, Cary Arthur

    1988-01-01

    Salicylhydroxamic acid (SHAM) stimulated germination of photosensitive lettuce (Lactuca sativa L. cv Waldmann's Green) seeds in darkness. To determine whether SHAM acts on the embryo or the endosperm, we investigated separately effects of SHAM on growth potential of isolated embryos as well as on endosperm strength. Embryo growth potential was quantified by incubating decoated embryos in various concentrations of osmoticum and measuring subsequent radicle elongation. Growth potential of embryos isolated from seeds pretreated with 4 millimolar SHAM was equal to that of untreated controls. Rupture strength of endosperm tissue excised from seeds pretreated with SHAM was 33% less than that of controls in the micropylar region. To determine if the embryo must be in contact with the endosperm for SHAM to weaken the endosperm, some endosperms were incubated with SHAM only after dissection from seeds. Rupture strength of SHAM-treated, isolated endosperms in the micropylar region was 25% less than that of untreated controls. There was no difference in rupture strength in the cotyledonary region of endosperm isolated from seeds treated with SHAM in buffer or buffer alone. SHAM therefore stimulates germination not by enhancing embryo growth potential, but by weakening the micropylar region of the endosperm enclosing the embryo. Images Fig. 1 PMID:11538237

  12. Ionic Strength Effect on the Rate of Reduction of Hexacyanoferrate(III) by Ascorbic Acid: A Flow Injection Kinetic Experiment

    NASA Astrophysics Data System (ADS)

    Nobrega, Joanquim A.; Rocha, Fabio R. P.

    1997-05-01

    Flow injection analysis (FIA) is a well recognized tool for solutions management. In spite of the use of this technique mainly for quantitative determination of analytes in solution, FIA systems can also be used for obtaining physical chemistry data. This work describes the use of a flow diagram to perform a kinetic experiment: the effect of ionic strength on the rate of reduction of hexacyanoferrate(III) by ascorbic acid. The rate determining step of this reaction involves the collision between two anionic species. The increase of the ionic strength of the medium alters the ionic atmosphere and changes the charge densities around the anions. Consequently, there is an increment of the rate constants for higher ionic strengths. In the proposed system, the flow is stopped by commutation when the center of the sample zone attained the flow cell and a gradual decrease in signal, related to the redox reaction, is registered as function of time. This allowed the determination of the rate constants as a function of the ionic strength. The product of the charges of the ions involved in the rate determining step was estimated in 3.2 that is close to the expected value considering the proposed mechanism.

  13. UV irradiation enhances the bonding strength between citric acid-crosslinked gelatin and stainless steel.

    PubMed

    Inoue, Motoki; Sasaki, Makoto; Katada, Yasuyuki; Taguchi, Tetsushi

    2011-11-01

    The effect of ultraviolet ray (UV) irradiation on the bonding strength between low carbon stainless steel 316 (SUS316L) and trisuccinimidyl citrate (TSC)-crosslinked alkali-treated gelatin (AlGelatin-TSC) was investigated. The UV irradiation effectively generated hydroxyl groups on the surface of SUS316L. The bonding strength between AlGelatin-TSC and SUS316L before UV irradiation was 0.345±0.007 MPa, and upon UV irradiation it increased to 0.750±0.069 MPa. In order to explain this enhanced bonding strength, the surface of SUS316L was examined using its water contact angle and X-ray photoelectron spectroscopy. Furthermore, the N 1s peaks derived from the TSC succinimidyl group were assigned to the surface of SUS316L after the immobilization of the TSC. This indicates that ester bond formation between the TSC active esters and the SUS316L hydroxyl groups contributed to the enhanced bonding strength. Therefore, UV irradiation and subsequent TSC immobilization is a simple way to functionalize biometal surfaces with various structures. This has practical applications for medical devices such as drug-eluting stents, dental implants, and metallic artificial bone.

  14. Interaction of metal ions with acid sites of biosorbents peat moss and Vaucheria and model substances alginic and humic acids

    SciTech Connect

    Crist, R.H.; Martin, J.R.; Crist, D.R.

    1999-07-01

    The interaction between added metal ions and acid sites of two biosorbents, peat moss and the alga Vaucheria, was studied. Results were interpreted in terms of two model substances, alginic acid, a copolymer of guluronic and mannuronic acids present in marine algae, and humic acid in peat moss. For peat moss and Vaucheria at pH 4--6, two protons were displaced per Cd sorbed, after correction for sorbed metals also displaced by the heavy metal. The frequent neglect of exchange of heavy metals for metals either sorbed on the native material or added for pH adjustment leads to erroneous conclusions about proton displacement stoichiometry. Proton displacement constants K{sub ex}{sup H} decreased logarithmically with pH and had similar slopes for alginic acid and biosorbents. This pH effect was interpreted as an electrostatic effect of increasing anionic charge making proton removal less favorable. The maximum number of exchangeable acid sites (capacity C{sub H}) decreased with pH for alginic acid but increased with pH for biosorbents. Consistent with titration behavior, this difference was explained in terms of more weak acid sites in the biosorbents.

  15. Effects of ionic strength and fulvic acid on adsorption of Tb(III) and Eu(III) onto clay

    NASA Astrophysics Data System (ADS)

    Poetsch, Maria; Lippold, Holger

    2016-09-01

    High salinity and natural organic matter are both known to facilitate migration of toxic or radioactive metals in geochemical systems, but little is known on their combined effect. We investigated complexation of Tb(III) and Eu(III) (as analogues for trivalent actinides) with fulvic acid and their adsorption onto a natural clay in the presence of NaCl, MgCl2 and CaCl2 up to very high ionic strengths. 160Tb, 152Eu and 14C-labelled fulvic acid were employed as radiotracers, allowing investigations at very low concentrations according to probable conditions in far-field scenarios of nuclear waste repositories. A combined Kd approach (Linear Additive Model) was tested for suitability in predicting solid-liquid distribution of metals in the presence of organic matter based on the interactions in the constituent subsystems. In this analysis, it could be shown that high ionic strength does not further enhance the mobilizing potential of humic matter. A quantitative reproduction of the influence of fulvic acid failed for most systems under study. Assumptions and limitations of the model are discussed.

  16. Effects of ionic strength and fulvic acid on adsorption of Tb(III) and Eu(III) onto clay.

    PubMed

    Poetsch, Maria; Lippold, Holger

    2016-09-01

    High salinity and natural organic matter are both known to facilitate migration of toxic or radioactive metals in geochemical systems, but little is known on their combined effect. We investigated complexation of Tb(III) and Eu(III) (as analogues for trivalent actinides) with fulvic acid and their adsorption onto a natural clay in the presence of NaCl, MgCl2 and CaCl2 up to very high ionic strengths. (160)Tb, (152)Eu and (14)C-labelled fulvic acid were employed as radiotracers, allowing investigations at very low concentrations according to probable conditions in far-field scenarios of nuclear waste repositories. A combined Kd approach (Linear Additive Model) was tested for suitability in predicting solid-liquid distribution of metals in the presence of organic matter based on the interactions in the constituent subsystems. In this analysis, it could be shown that high ionic strength does not further enhance the mobilizing potential of humic matter. A quantitative reproduction of the influence of fulvic acid failed for most systems under study. Assumptions and limitations of the model are discussed. PMID:27454893

  17. Hydrogen adsorption strength and sites in the metal organic framework MOF5: Comparing experiment and model calculations

    NASA Astrophysics Data System (ADS)

    Mulder, F. M.; Dingemans, T. J.; Schimmel, H. G.; Ramirez-Cuesta, A. J.; Kearley, G. J.

    2008-07-01

    Hydrogen adsorption in porous, high surface area, and stable metal organic frameworks (MOF's) appears a novel route towards hydrogen storage materials [N.L. Rosi, J. Eckert, M. Eddaoudi, D.T. Vodak, J. Kim, M. O'Keeffe, O.M. Yaghi, Science 300 (2003) 1127; J.L.C. Rowsell, A.R. Millward, K. Sung Park, O.M. Yaghi, J. Am. Chem. Soc. 126 (2004) 5666; G. Ferey, M. Latroche, C. Serre, F. Millange, T. Loiseau, A. Percheron-Guegan, Chem. Commun. (2003) 2976; T. Loiseau, C. Serre, C. Huguenard, G. Fink, F. Taulelle, M. Henry, T. Bataille, G. Férey, Chem. Eur. J. 10 (2004) 1373]. A prerequisite for such materials is sufficient adsorption interaction strength for hydrogen adsorbed on the adsorption sites of the material because this facilitates successful operation under moderate temperature and pressure conditions. Here we report detailed information on the geometry of the hydrogen adsorption sites, based on the analysis of inelastic neutron spectroscopy (INS). The adsorption energies for the metal organic framework MOF5 equal about 800 K for part of the different sites, which is significantly higher than for nanoporous carbon materials (˜550 K) [H.G. Schimmel, G.J. Kearley, M.G. Nijkamp, C.T. Visser, K.P. de Jong, F.M. Mulder, Chem. Eur. J. 9 (2003) 4764], and is in agreement with what is found in first principles calculations [T. Sagara, J. Klassen, E. Ganz, J. Chem. Phys. 121 (2004) 12543; F.M. Mulder, T.J. Dingemans, M. Wagemaker, G.J. Kearley, Chem. Phys. 317 (2005) 113]. Assignments of the INS spectra is realized using comparison with independently published model calculations [F.M. Mulder, T.J. Dingemans, M. Wagemaker, G.J. Kearley, Chem. Phys. 317 (2005) 113] and structural data [T. Yildirim, M.R. Hartman, Phys. Rev. Lett. 95 (2005) 215504].

  18. A study of the source-receptor relationships influencing the acidity of precipitation collected at a rural site in France

    NASA Astrophysics Data System (ADS)

    Charron, Aurélie; Plaisance, Hervé; Sauvage, Stéphane; Coddeville, Patrice; Galloo, Jean-Claude; Guillermo, René

    In order to examine the qualitative and quantitative source-receptor relationships responsible for acid rains at a background site in France, a receptor-oriented model was applied to the precipitation data collected from 1992 to 1995. Origins of acidic and alkaline species in precipitations have been investigated. The methodology combines precipitation chemical data with air parcel backward trajectories to establish concentration field maps of likely contributing sources. Highest acidities and concentrations of sulfate and nitrate in precipitation were associated with transport from the high emission areas of central Europe. Alkaline events were associated with air masses originating from Mediterranean basin or northern Africa. The quantitative relationships between the maps of potential sources and the European emissions of SO 2 and NO x were examined performing a correlation analysis. Good correlations were found between computed concentrations of acidic species and emissions of SO 2 and NO x. Substantial seasonal variations of acidic species were revealed. The highest concentrations occurred during the warm season. These seasonal variations are the effect of change of meteorological conditions and of the strength atmospheric processes according to the season.

  19. Modeling solubility, acid-base properties and activity coefficients of amoxicillin, ampicillin and (+)6-aminopenicillanic acid, in NaCl(aq) at different ionic strengths and temperatures.

    PubMed

    Crea, Francesco; Cucinotta, Daniela; De Stefano, Concetta; Milea, Demetrio; Sammartano, Silvio; Vianelli, Giuseppina

    2012-11-20

    The total solubility of three penicillin derivatives was determined, in pure water and NaCl aqueous solutions at different salt concentrations (from ∼0.15 to 1.0 mol L(-1) for ampicillin and amoxicillin, and from ∼0.05 to 2.0 mol L(-1) for (+)6-aminopenicillanic acid), using the shake-flask method for generating the saturated solutions, followed by potentiometric analysis. The knowledge of the pH of solubilization and of the protonation constants determined in the same experimental conditions, allowed us to calculate, by means of the mass balance equations, the solubility of the neutral species at different ionic strength values, to model its dependence on the salt concentration and to determine the corresponding values at infinite dilution. The salting parameter and the activity coefficients of the neutral species were calculated by the Setschenow equation. The protonation constants of ampicillin and amoxicillin, determined at different temperatures (from T=288.15 to 318.15K), from potentiometric and spectrophotometric measurements, were used to calculate, by means of the Van't Hoff equation, the temperature coefficients at different ionic strength values and the corresponding protonation entropies. The protonation enthalpies of the (+)6-aminopenicillanic acid were determined by isoperibol calorimetric titrations at T=298.15K and up to I=2.0 mol L(-1). The dependence of the protonation constants on ionic strength was modeled by means of the Debye-Hückel and SIT (Specific ion Interaction Theory) approaches, and the specific interaction parameters of the ionic species were determined. The hydrolysis of the β-lactam ring was studied by spectrophotometric and H NMR investigations as a function of pH, ionic strength and time. Potentiometric measurements carried out on the hydrolyzed (+)6-aminopenicillanic acid allowed us to highlight that the opened and the closed β-lactam forms of the (+)6-aminopenicillanic acid have quite different acid-base properties. An

  20. Effects of egg white protein supplementation on muscle strength and serum free amino acid concentrations.

    PubMed

    Hida, Azumi; Hasegawa, Yuko; Mekata, Yuko; Usuda, Mika; Masuda, Yasunobu; Kawano, Hitoshi; Kawano, Yukari

    2012-10-01

    The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM), one repetition maximum (1RM) muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal) and carbohydrate group (17.5 g maltodextrin, 78 kcal). Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press) increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength.

  1. Effect of storage and acid etching on the tensile bond strength of composite resins to glass ionomer cement.

    PubMed

    Mesquita, M F; Domitti, S S; Consani, S; de Goes, M F

    1999-01-01

    This in vitro study evaluates the effect of storage time and acid etching on the tensile bond strength of glass ionomer cement to composite resins. The bonded assemblies were stored at 100% relative humidity and 37 degrees C for 1 hour, 1 day, 1 week, 1 month and 3 months. The test specimen was loaded at tension to failure on an Otto Wolpert-Werke testing instrument with a crosshead speed of 6 mm/min. The results showed a significant statistical difference for etched Vidrion F when compared to etched Ketac Bond at all storage periods. The unetched samples were statistically similar at 3 months, with the highest values for Vidrion F.

  2. Site-Specific Advantages in Skeletal Geometry and Strength at the Proximal Femur and Forearm in Young Female Gymnasts

    PubMed Central

    Dowthwaite, Jodi N.; Rosenbaum, Paula F.; Scerpella, Tamara A.

    2012-01-01

    Purpose We evaluated site-specific skeletal adaptation to loading during growth,comparing radius (RAD) and femoral neck (FN) DXA scans in young female gymnasts (GYM) and non-gymnasts (NON). Methods Subjects from an ongoing longitudinal study (8-26 yrs old) underwent annual DXA scans (proximal femur, forearm, total body) and anthropometry, completing maturity and physical activity questionnaires. This cross-sectional analysis used the most recent data meeting the following criteria: gynecological age ≤2.5 yrs post-menarche; GYM annual mean gymnastic exposure ≥5.0 h/wk in the prior year. Bone geometric and strength indices were derived from scans for 173 subjects (8-17 yrs old) via hip structural analysis (femoral narrow neck, NN) and similar radius formulae (1/3 and Ultradistal (UD)). Maturity was coded as M1 (Tanner I breast), M2 (pre-menarche, ≥Tanner II breast) or M3 (post-menarche). ANOVA and chi square compared descriptive data. Two factor ANCOVA adjusted for age, height, total body non-bone lean mass and percent body fat; significance was tested for main effects and interactions between gymnastic exposure and maturity. Results At the distal radius, GYM means were significantly greater than NON means for all variables (p<0.05). At the proximal femur, GYM exhibited narrower periosteal and endosteal dimensions, but greater indices of cortical thickness, BMC, aBMD and section modulus, with lower buckling ratio (p <0.05). However, significant interactions between maturity and loading were detected for the following: 1) FN bone mineral content (BMC), NN buckling ratio (GYM BMC advantages only in M1 and M3; for BMC and buckling ratio, M1 advantages were greatest; 2) 1/3 radius BMC, width, endosteal diameter, cortical cross-sectional area, section modulus (GYM advantages primarily post-menarche); 3) UD radius BMC and axial compressive strength (GYM advantages were larger with greater maturity, greatest post-menarche). Conclusions Maturity-specific comparisons

  3. Influence of different forms of acidities on soil microbiological properties and enzyme activities at an acid mine drainage contaminated site.

    PubMed

    Sahoo, Prafulla Kumar; Bhattacharyya, Pradip; Tripathy, Subhasish; Equeenuddin, Sk Md; Panigrahi, M K

    2010-07-15

    Assessment of microbial parameters, viz. microbial biomass, fluorescence diacetate, microbial respiration, acid phosphatase, beta-glucosidase and urease with respect to acidity helps in evaluating the quality of soils. This study was conducted to investigate the effects of different forms of acidities on soil microbial parameters in an acid mine drainage contaminated site around coal deposits in Jainta Hills of India. Total potential and exchangeable acidity, extractable and exchangeable aluminium were significantly higher in contaminated soil compared to the baseline (p<0.01). Different forms of acidity were significantly and positively correlated with each other (p<0.05). Further, all microbial properties were positively and significantly correlated with organic carbon and clay (p<0.05). The ratios of microbial parameters with organic carbon were negatively correlated with different forms of acidity. Principal component analysis and cluster analyses showed that the microbial activities are not directly influenced by the total potential acidity and extractable aluminium. Though acid mine drainage affected soils had higher microbial biomass and activities due to higher organic matter content than those of the baseline soils, the ratios of microbial parameters/organic carbon indicated suppression of microbial growth and activities due to acidity stress. PMID:20417031

  4. Evidence that Chemical Chaperone 4-Phenylbutyric Acid Binds to Human Serum Albumin at Fatty Acid Binding Sites

    PubMed Central

    James, Joel; Shihabudeen, Mohamed Sham; Kulshrestha, Shweta; Goel, Varun; Thirumurugan, Kavitha

    2015-01-01

    Endoplasmic reticulum stress elicits unfolded protein response to counteract the accumulating unfolded protein load inside a cell. The chemical chaperone, 4-Phenylbutyric acid (4-PBA) is a FDA approved drug that alleviates endoplasmic reticulum stress by assisting protein folding. It is found efficacious to augment pathological conditions like type 2 diabetes, obesity and neurodegeneration. This study explores the binding nature of 4-PBA with human serum albumin (HSA) through spectroscopic and molecular dynamics approaches, and the results show that 4-PBA has high binding specificity to Sudlow Site II (Fatty acid binding site 3, subdomain IIIA). Ligand displacement studies, RMSD stabilization profiles and MM-PBSA binding free energy calculation confirm the same. The binding constant as calculated from fluorescence spectroscopic studies was found to be kPBA = 2.69 x 105 M-1. Like long chain fatty acids, 4-PBA induces conformational changes on HSA as shown by circular dichroism, and it elicits stable binding at Sudlow Site II (fatty acid binding site 3) by forming strong hydrogen bonding and a salt bridge between domain II and III of HSA. This minimizes the fluctuation of HSA backbone as shown by limited conformational space occupancy in the principal component analysis. The overall hydrophobicity of W214 pocket (located at subdomain IIA), increases upon occupancy of 4-PBA at any FA site. Descriptors of this pocket formed by residues from other subdomains largely play a role in compensating the dynamic movement of W214. PMID:26181488

  5. Lots of data, how do we use it? Strengths and inaccuracies of utility acid rain electronic data reports

    SciTech Connect

    Schott, J.

    1996-12-31

    Entergy Corporation is a Phase II utility with a fossil generation base composed primarily natural gas and low sulfur coal. This paper presents an analysis of a large Phase II utility`s continuous emissions monitoring data reported to EPA under Title IV Acid Rain. Electric utilities currently report hourly emissions of NOx, SO{sub 2}, CO{sub 2}, fuel use, and generation through electronic data reports to EPA. This paper describes strengths and weaknesses of the data reported to EPA as determined through an analysis of 1995 data. Emissions reported by this company tinder acid rain for SO{sub 2} and NOx are very different from emissions reported to state agencies for annual emission inventory purposes in past years and will represent a significant break with historic trends. A comparison of emissions has been made of 1995 emissions reported under Electronic Data Reports to the emissions that would have been reported using emission factors and fuel data in past years. In addition, the paper examines the impacts of 40 CFR Part 75 Acid Rain requirements such as missing data substitution and monitor bias adjustments. Measurement system errors including stack flow measurement and false NOx Lb/MMBtu readings at very low loads are discussed. This paper describes the implications for public policy, compliance, emissions inventories, and business decisions of Part 75 acid rain monitoring and reporting requirements.

  6. High melt strength, tear resistant blown film based on poly(lactic acid)

    NASA Astrophysics Data System (ADS)

    Edmonds, Neil R.; Plimmer, Peter N.; Tanner, Chris

    2015-05-01

    A major problem associated with the commercial manufacture of thin films from PLA is inferior processing characteristics on blown film lines compared to low density polyethylene. PLA has poor melt strength (leading to bubble instability) and develops a permanent crease in the flattened film as it exits the tower of the film line. In addition, the thin film product has poor tear strength and an unacceptable `noise' level when converted into flexible packaging. Furthermore, fabricated articles based on PLA are known to show an unattractive tendency toward dimensional instability. This behaviour is associated with `cold crystallization', a phenomenon which also causes exudation of any plasticizer added for improving flexibility. Blow moulded articles based on PLA also exhibit dimensional sensitivity above 60°C. All of these issues have been overcome by the technology described in this paper. This has been accomplished without loss of the valuable compostability characteristic of PLA; this was confirmed by evaluation of film in a commercial composting operation. These results have been achieved through novel reactive compounding technology which: (a) Creates a PLA-rich structure containing long chain crosslinks, (b) generates a low glass transition temperature phase covalently bonded to the PLA structure, and (c) provides a material which performs like LDPE in a blown film manufacturing operation. The technology developed is covered by NZ Patent 580231 (3). The patent is held by UniServices Ltd, The University of Auckland, New Zealand.

  7. Acid-base site detection and mapping on solid surfaces by Kelvin force microscopy (KFM).

    PubMed

    Gouveia, Rubia F; Bernardes, Juliana S; Ducati, Telma R D; Galembeck, Fernando

    2012-12-01

    Electrostatic potential at the surface of acidic or basic solids changes under higher relative humidity (RH), as determined by using Kelvin force microscopy (KFM). The potential on acid surfaces becomes more negative as the water vapor pressure increases, while it becomes more positive on basic solids. These results verify the following hypothesis: OH(-) or H(+) ions associated with atmospheric water ion clusters are selectively adsorbed on solid surfaces, depending on the respective Brønsted acid or base character. Therefore, Kelvin microscopy, under variable humidity, is a rigorous but convenient alternative to determine the acid-base character of solid surfaces, with a great advantage: it uses only one amphoteric and simple reagent to determine both the acid and base sites. Moreover, this technique provides information on the spatial distribution of acid-base sites, which is currently inaccessible to any other method.

  8. Soybean root growth in acid subsoils in relation to magnesium additions and soil solution ionic strength

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hydroponic studies with soybean [Glycine max (L.) Merr.] have shown that µM additions of Mg2+ were as effective in ameliorating Al rhizotoxicity as additions of Ca2+ in the mM concentration range. The objectives of this study were to assess ameliorative effects of Mg on soybean root growth in acidic...

  9. Probing the Binding Site of Bile Acids in TGR5.

    PubMed

    Macchiarulo, Antonio; Gioiello, Antimo; Thomas, Charles; Pols, Thijs W H; Nuti, Roberto; Ferrari, Cristina; Giacchè, Nicola; De Franco, Francesca; Pruzanski, Mark; Auwerx, Johan; Schoonjans, Kristina; Pellicciari, Roberto

    2013-12-12

    TGR5 is a G-protein-coupled receptor (GPCR) mediating cellular responses to bile acids (BAs). Although some efforts have been devoted to generate homology models of TGR5 and draw structure-activity relationships of BAs, none of these studies has hitherto described how BAs bind to TGR5. Here, we present an integrated computational, chemical, and biological approach that has been instrumental to determine the binding mode of BAs to TGR5. As a result, key residues have been identified that are involved in mediating the binding of BAs to the receptor. Collectively, these results provide new hints to design potent and selective TGR5 agonists. PMID:24900622

  10. Probing the Binding Site of Bile Acids in TGR5

    PubMed Central

    2013-01-01

    TGR5 is a G-protein-coupled receptor (GPCR) mediating cellular responses to bile acids (BAs). Although some efforts have been devoted to generate homology models of TGR5 and draw structure–activity relationships of BAs, none of these studies has hitherto described how BAs bind to TGR5. Here, we present an integrated computational, chemical, and biological approach that has been instrumental to determine the binding mode of BAs to TGR5. As a result, key residues have been identified that are involved in mediating the binding of BAs to the receptor. Collectively, these results provide new hints to design potent and selective TGR5 agonists. PMID:24900622

  11. Organic acids in cloud water and rainwater at a mountain site in acid rain areas of South China.

    PubMed

    Sun, Xiao; Wang, Yan; Li, Haiyan; Yang, Xueqiao; Sun, Lei; Wang, Xinfeng; Wang, Tao; Wang, Wenxing

    2016-05-01

    To investigate the chemical characteristics of organic acids and to identify their source, cloud water and rainwater samples were collected at Mount Lu, a mountain site located in the acid rain-affected area of south China, from August to September of 2011 and March to May of 2012. The volume-weighted mean (VWM) concentration of organic acids in cloud water was 38.42 μeq/L, ranging from 7.45 to 111.46 μeq/L, contributing to 2.50 % of acidity. In rainwater samples, organic acid concentrations varied from 12.39 to 68.97 μeq/L (VWM of 33.39 μeq/L). Organic acids contributed significant acidity to rainwater, with a value of 17.66 %. Formic acid, acetic acid, and oxalic acid were the most common organic acids in both cloud water and rainwater. Organic acids had an obviously higher concentration in summer than in spring in cloud water, whereas there was much less discrimination in rainwater between the two seasons. The contribution of organic acids to acidity was lower during summer than during spring in both cloud water (2.20 % in summer vs 2.83 % in spring) and rainwater (12.24 % in summer vs 19.89 % in spring). The formic-to-acetic acid ratio (F/A) showed that organic acids were dominated by primary emissions in 71.31 % of the cloud water samples and whole rainwater samples. Positive matrix factorization (PMF) analysis determined four factors as the sources of organic acids in cloud water, including biogenic emissions (61.8 %), anthropogenic emissions (15.28 %), marine emissions (15.07 %) and soil emissions (7.85 %). The findings from this study imply an indispensable role of organic acids in wet deposition, but organic acids may have a limited capacity to increase ecological risks in local environments. PMID:26841776

  12. Organic acids in cloud water and rainwater at a mountain site in acid rain areas of South China.

    PubMed

    Sun, Xiao; Wang, Yan; Li, Haiyan; Yang, Xueqiao; Sun, Lei; Wang, Xinfeng; Wang, Tao; Wang, Wenxing

    2016-05-01

    To investigate the chemical characteristics of organic acids and to identify their source, cloud water and rainwater samples were collected at Mount Lu, a mountain site located in the acid rain-affected area of south China, from August to September of 2011 and March to May of 2012. The volume-weighted mean (VWM) concentration of organic acids in cloud water was 38.42 μeq/L, ranging from 7.45 to 111.46 μeq/L, contributing to 2.50 % of acidity. In rainwater samples, organic acid concentrations varied from 12.39 to 68.97 μeq/L (VWM of 33.39 μeq/L). Organic acids contributed significant acidity to rainwater, with a value of 17.66 %. Formic acid, acetic acid, and oxalic acid were the most common organic acids in both cloud water and rainwater. Organic acids had an obviously higher concentration in summer than in spring in cloud water, whereas there was much less discrimination in rainwater between the two seasons. The contribution of organic acids to acidity was lower during summer than during spring in both cloud water (2.20 % in summer vs 2.83 % in spring) and rainwater (12.24 % in summer vs 19.89 % in spring). The formic-to-acetic acid ratio (F/A) showed that organic acids were dominated by primary emissions in 71.31 % of the cloud water samples and whole rainwater samples. Positive matrix factorization (PMF) analysis determined four factors as the sources of organic acids in cloud water, including biogenic emissions (61.8 %), anthropogenic emissions (15.28 %), marine emissions (15.07 %) and soil emissions (7.85 %). The findings from this study imply an indispensable role of organic acids in wet deposition, but organic acids may have a limited capacity to increase ecological risks in local environments.

  13. Site and chirality selective chemical modifications of boron nitride nanotubes (BNNTs) via Lewis acid-base interactions.

    PubMed

    Sundaram, Rajashabala; Scheiner, Steve; Roy, Ajit K; Kar, Tapas

    2015-02-01

    The pristine BNNTs contain both Lewis acid (boron) and Lewis base (nitrogen) centers at their surface. Interactions of ammonia and borane molecules, representatives of Lewis base and acid as adsorbates respectively, with matching sites at the surface of BNNTs, have been explored in the present DFT study. Adsorption energies suggest stronger chemisorption (about 15-20 kcal mol(-1)) of borane than ammonia (about 5-10 kcal mol(-1)) in both armchair (4,4) and zigzag (8,0) variants of the tube. NH3 favors (8,0) over the (4,4) tube, whereas BH3 exhibits the opposite preference, indicating some chirality dependence on acid-base interactions. A new feature of bonding is found in BH3/AlH3-BNNTs (at the edge site) complexes, where one hydrogen of the guest molecule is involved in three-center two-electron bonding, in addition to dative covalent bond (N: → B). This interaction causes a reversal of electron flow from borane/alane to BNNT, making the tube an electron acceptor, suggesting tailoring of electronic properties could be possible by varying strength of incoming Lewis acids. On the contrary, BNNTs always behave as electron acceptor in ammonia complexes. IR, XPS and NMR spectra show some characteristic features of complexes and can help experimentalists to identify not only structures of such complexes but also the location of the guest molecules and design second functionalizations. Interaction with several other neutral BF3, BCl3, BH2CH3 and ionic CH3(+) acids as well as amino group (CH3NH2 and NH2COOH) were also studied. The strongest interaction (>100 kcal mol(-1)) is found in BNNT-CH3(+) complexes and H-bonds are the only source of stability of NH2COOH-BNNT complexes. PMID:25559141

  14. Interaction morphology and bond strength of nanofilled simplified-step adhesives to acid etched dentin

    PubMed Central

    Di Hipólito, Vinicius; Reis, André Figueiredo; Mitra, Sumita B.; de Goes, Mario Fernando

    2012-01-01

    Objective: To evaluate the effect of nanofillers incorporated into adhesives on the microtensile bond strength (μ-TBS) and interfacial micromorphology to dentin. Methods: The occlusal enamel of 5 human molars was removed and each tooth sectioned into four quarters. The exposed dentin was treated with one of the following adhesives: Adper Single Bond (SB-unfilled), OptiBond Solo Plus (OS-barium aluminoborosilicate, 400nm Ø), Prime & Bond NT (NT-colloidal silica, 7–40 nm Ø) and Adper Single Bond 2 (SB2-colloidal silica, 5nm Ø). Cylinders of resin-based composite were constructed on the adhesive layers. After 24-hour storage, the restored tooth-quadrants were sectioned to obtain stick-shaped specimens (0.8 mm2, cross-sectional area) and submitted to μ-TBS at a cross-speed of 0.5 mm/min. Data were analyzed using one-way ANOVA and Tukey’s test (alpha = .05). Twenty-eight additional teeth were used for interfacial micro-morphologic analysis by SEM (16-teeth) and TEM (12-teeth). The dentin surfaces of 32 discs were treated with the adhesives (8 discs for adhesive) and laminated to form disc-pairs using a flowable resin composite for SEM/EDS analysis. For TEM, 90nm-thick nondemineralized unstained sections were processed. Results: SB2 showed significant higher bond strength than SB, OS and NT. The SEM/EDS and TEM analysis revealed nanofillers infiltrated within the interfibrillar spaces of the SB2-hybrid layer. Fillers were concentrated around patent tubular orifices and in the adhesive layer for OS and NT. Conclusion: The presence of nanofillers within the interfibrillar spaces of the SB2-hybrid layer suggests its importance in the improvement of the μ-TBS. PMID:23077413

  15. Effect of the acidic strength on the vapor phase Beckmann rearrangement of cyclohexanone oxime over the MFI zeolite: an embedded ONIOM study.

    PubMed

    Sirijaraensre, Jakkapan; Limtrakul, Jumras

    2009-01-21

    The mechanism and energetic profile of the Beckmann rearrangement reaction of cyclohexanone oxime to epsilon-caprolactam catalyzed by the H-[Al]-MFI and H-[B]-MFI zeolites were investigated by both the bare cluster and the ONIOM models at the B3LYP/6-31G(d,p) and the B3LYP/6-31G(d,p):MNDO levels of theory, respectively. In order to improve the energetic properties and take into account the whole zeolite framework effect, single point calculations are undertaken at the embedded ONIOM2 schemes; MP2/6-311G(d,p):HF/6-31G(d) with an additional long-range electrostatic potential from the extended zeolite framework. The reaction mechanism of the Beckmann rearrangement over the acid site of zeolites consists of three steps: the 1,2 H shift, the rearrangement and the tautomerization. The activation energies for the Beckmann rearrangement of cyclohexanone oxime on the H-[Al]-MFI zeolite are calculated to be 31.46, 16.15 and 18.95 kcal mol(-1), for the first, second and third steps, respectively, whereas in the H-[B]-MFI zeolite, the energy barriers for each step of the reaction are 24.33, 7.46 and 20.43 kcal mol(-1), respectively. The rate-determining step of the reaction is the first step, which is the transformation from the N-ended cyclohexanone oxime adsorption complex and the O-ended one. These results signify the important role that the acid strength of zeolites plays in altering the energy profile of the reaction. The results further indicate that the weak Brønsted acid sites in the [B]-MFI zeolite could better catalyze the Beckmann rearrangement of cyclohexanone oxime than the strong acid sites in the [Al]-MFI zeolite, as compared with the quantitatively low activation energy of most steps. However, the turnover reaction of the H-[B]-MFI zeolite might be delayed by the quantitatively high desorption energy of the product as compared to the adsorption energy of the reactant.

  16. A comparative study of shear bond strength of orthodontic bracket after acid-etched and Er:YAG treatment on enamel surface

    NASA Astrophysics Data System (ADS)

    Leão, Juliana C.; Mota, Cláudia C. B. O.; Cassimiro-silva, Patricia F.; Gomes, Anderson S. L.

    2016-02-01

    This study aimed to evaluate the shear bond strength (SBS) of teeth prepared for orthodontic bracket bonding with 37% phosphoric acid and Er:YAG laser. Forty bovine incisors were divided into two groups. In Group I, the teeth were conditioned with 37% phosphoric acid and brackets were bonded with Transbond XT; in Group II, the teeth were irradiated with Er:YAG and bonding with Transbond XT. After SBS test, the adhesive remnant index was determined. Adhesion to dental hard tissues after Er:YAG laser etching was inferior to that obtained after acid etching but exceeded what is believed to be clinically sufficient strength, and therefore can be used in patients.

  17. Shear bond strength of metal brackets to feldspathic porcelain treated by Nd:YAG laser and hydrofluoric acid.

    PubMed

    Hosseini, Mohammad Hashem; Sobouti, Farhad; Etemadi, Ardavan; Chiniforush, Nasim; Shariati, Mahsa

    2015-02-01

    Adult orthodontic treatment requires bonding orthodontic attachment to dental restorations. Ceramics are commonly used as esthetic restorative materials for the crowns and bridges. The present study evaluated the shear bond strength of metal orthodontic brackets to the feldspathic porcelain surfaces following conditioning by different powers of neodymium-doped yttrium aluminum garnet (Nd:YAG) laser and hydrofluoric acid as a conventional method. Seventy-two glazed porcelain samples were prepared and randomly attributed to six equal groups of 12. In the conventional hydrofluoric (HF) group, the specimens were etched by 9.6% hydrofluoric acid for 4 min. In laser groups, samples were conditioned by 0.75-, 1-, 1.25-, 1.5-, and 2-W Nd:YAG laser for 10 s. Metal brackets were bonded to porcelain samples and after being stored in distilled water for 24 h, they were subjected to thermocycling for 500 cycles. The debonding was carried out by a Zwick testing machine. The data were statistically analyzed by ANOVA and Tamhane multiple comparisons tests. The mean ± SD of the shear bond strength in the laser group 0.75, 1, 1.25, 1.5, and 2 W and HF group was 2.2 ± 0.9, 4.2 ± 1.1, 4.9 ± 2.4, 7 ± 1.7, 9.6 ± 2.7, and 9.4 ± 2.5, respectively. Together with the increased power of laser, the mean shear bond strength was increased continuously and no significant differences were found between the HF group and the laser groups with power of 1.5 or 2 W. Also, there was no significant difference between all test groups in ARI scores. There was no significant difference between bond strength of laser groups with power of 1.5 and 2 W and HF-etched group. So, Nd:YAG laser with appropriate parameters can be used as an alternative method for porcelain etching.

  18. Na+ Inhibits the Epithelial Na+ Channel by Binding to a Site in an Extracellular Acidic Cleft*

    PubMed Central

    Kashlan, Ossama B.; Blobner, Brandon M.; Zuzek, Zachary; Tolino, Michael; Kleyman, Thomas R.

    2015-01-01

    The epithelial Na+ channel (ENaC) has a key role in the regulation of extracellular fluid volume and blood pressure. ENaC belongs to a family of ion channels that sense the external environment. These channels have large extracellular regions that are thought to interact with environmental cues, such as Na+, Cl−, protons, proteases, and shear stress, which modulate gating behavior. We sought to determine the molecular mechanism by which ENaC senses high external Na+ concentrations, resulting in an inhibition of channel activity. Both our structural model of an ENaC α subunit and the resolved structure of an acid-sensing ion channel (ASIC1) have conserved acidic pockets in the periphery of the extracellular region of the channel. We hypothesized that these acidic pockets host inhibitory allosteric Na+ binding sites. Through site-directed mutagenesis targeting the acidic pocket, we modified the inhibitory response to external Na+. Mutations at selected sites altered the cation inhibitory preference to favor Li+ or K+ rather than Na+. Channel activity was reduced in response to restraining movement within this region by cross-linking structures across the acidic pocket. Our results suggest that residues within the acidic pocket form an allosteric effector binding site for Na+. Our study supports the hypothesis that an acidic cleft is a key ligand binding locus for ENaC and perhaps other members of the ENaC/degenerin family. PMID:25389295

  19. Na+ inhibits the epithelial Na+ channel by binding to a site in an extracellular acidic cleft.

    PubMed

    Kashlan, Ossama B; Blobner, Brandon M; Zuzek, Zachary; Tolino, Michael; Kleyman, Thomas R

    2015-01-01

    The epithelial Na(+) channel (ENaC) has a key role in the regulation of extracellular fluid volume and blood pressure. ENaC belongs to a family of ion channels that sense the external environment. These channels have large extracellular regions that are thought to interact with environmental cues, such as Na(+), Cl(-), protons, proteases, and shear stress, which modulate gating behavior. We sought to determine the molecular mechanism by which ENaC senses high external Na(+) concentrations, resulting in an inhibition of channel activity. Both our structural model of an ENaC α subunit and the resolved structure of an acid-sensing ion channel (ASIC1) have conserved acidic pockets in the periphery of the extracellular region of the channel. We hypothesized that these acidic pockets host inhibitory allosteric Na(+) binding sites. Through site-directed mutagenesis targeting the acidic pocket, we modified the inhibitory response to external Na(+). Mutations at selected sites altered the cation inhibitory preference to favor Li(+) or K(+) rather than Na(+). Channel activity was reduced in response to restraining movement within this region by cross-linking structures across the acidic pocket. Our results suggest that residues within the acidic pocket form an allosteric effector binding site for Na(+). Our study supports the hypothesis that an acidic cleft is a key ligand binding locus for ENaC and perhaps other members of the ENaC/degenerin family. PMID:25389295

  20. Comparative Evaluation of Tensile – Bond Strength of An Orthodontic Adhesive with and without Fluoride Application, After Acid Etching -An Invitro Study

    PubMed Central

    Yugandhar, G; Ramana, I Venkata; Srinivas, K; Yadav, S. Sarjeev Singh

    2015-01-01

    Background Fixed appliances hinder the effective control of plaque accumulation and white spot lesions may develop under the ill fitting bands or adjacent to the stainless steel brackets during orthodontic treatment particularly the etching process. Aims and Objectives Comparative study of tensile bond strength of an orthodontic adhesive with and without fluoride application after acid etching to know the effect of fluoride on bond strength. Materials and Methods This study is carried out on 90 non carious human premolar teeth, and divided in 6 groups with each group of 15 specimens. In those Groups I and IV were control group acid etch treatment, Group II and V is 1.23% APF gel (acid etch plus APF gel treatment,) and group III and VI is 8% SnF2 (acid etch plus SnF2 treatment). Samples of Group I, II and III bond strength were tested after 24 h and groups IV, V and VI after one month on microtechtensometer machine. The scanning electron microscope (SEM) investigation was carried out for the 2 specimens for the control group after acid etch and 4 specimens after acid etch with fluoride application for fluoride groups. Results Control and SnF2 treated groups was found to be nearly similar to the control group whereas APF treated group showed less focal holes than the other 2 groups. Conclusion Fluoride application after acid etching without having an adverse effect on bond strength but we can prevent the white spot lesions and caries. PMID:26023648

  1. Site specific incorporation of heavy atom-containing unnatural amino acids into proteins for structure determination

    DOEpatents

    Xie, Jianming; Wang, Lei; Wu, Ning; Schultz, Peter G.

    2008-07-15

    Translation systems and other compositions including orthogonal aminoacyl tRNA-synthetases that preferentially charge an orthogonal tRNA with an iodinated or brominated amino acid are provided. Nucleic acids encoding such synthetases are also described, as are methods and kits for producing proteins including heavy atom-containing amino acids, e.g., brominated or iodinated amino acids. Methods of determining the structure of a protein, e.g., a protein into which a heavy atom has been site-specifically incorporated through use of an orthogonal tRNA/aminoacyl tRNA-synthetase pair, are also described.

  2. Improve the Strength of PLA/HA Composite Through the Use of Surface Initiated Polymerization and Phosphonic Acid Coupling Agent

    PubMed Central

    Wang, Tongxin; Chow, Laurence C.; Frukhtbeyn, Stanislav A.; Ting, Andy Hai; Dong, Quanxiao; Yang, Mingshu; Mitchell, James W.

    2011-01-01

    Bioresorbable composite made from degradable polymers, e.g., polylactide (PLA), and bioactive calcium phosphates, e.g., hydroxyapatite (HA), are clinically desirable for bone fixation, repair and tissue engineering because they do not need to be removed by surgery after the bone heals. However, preparation of PLA/HA composite from non-modified HA usually results in mechanical strength reductions due to a weak interface between PLA and HA. In this study, a calcium-phosphate/phosphonate hybrid shell was developed to introduce a greater amount of reactive hydroxyl groups onto the HA particles. Then, PLA was successfully grafted on HA by surface-initiated polymerization through the non-ionic surface hydroxyl groups. Thermogravimetric analysis indiated that the amount of grafted PLA on HA can be up to 7 %, which is about 50 % greater than that from the literature. PLA grafted HA shows significantly different pH dependent ζ-potential and particle size profiles from those of uncoated HA. By combining the phosphonic acid coupling agent and surface initiated polymerization, PLA could directly link to HA through covalent bond so that the interfacial interaction in the PLA/HA composite can be significantly improved. The diametral tensile strength of PLA/HA composite prepared from PLA-grafted HA was found to be over twice that of the composite prepared from the non-modified HA. Moreover, the tensile strength of the improved composite was 23 % higher than that of PLA alone. By varying additional variables, this approach has the potential to produce bioresorbable composites with improved mechanical properties that are in the range of natural bones, and can have wide applications for bone fixation and repair in load-bearing areas. PMID:22399838

  3. Hydrogen-induced embrittlement wear of a high-strength, low-alloy steel in an acidic environment

    SciTech Connect

    Zhang, T.C.; Jiang, X.X.; Li, S.Z.

    1997-03-01

    Corrosive wear of a high-strength, low-alloy steel (HSLA) was examined in 0.02 mol/L sulfuric acid (H{sub 2}SO{sub 4}) solution at different polarized potentials and loads using a pin-on-disc wear device and a potentiostat. Morphologies of the wear tracks were observed by scanning electron microscopy (SEM). Hydrogen content in the surface or subsurface of wear tracks was determined using secondary ion mass spectrometry (SIMS). Results showed the increased material removal with a negative of potential in the cathodically polarized range resulted from the synergistic effect of hydrogen-induced damage and mechanical wear from hydrogen evolution on the wear surface. Increases in wear loss with potential in the anodically polarized range resulted from synergism between anodic dissolution and wear.

  4. Effect of hydrofluoric acid etching on shear bond strength of an indirect resin composite to an adhesive cement.

    PubMed

    Hori, Sayaka; Minami, Hiroyuki; Minesaki, Yoshito; Matsumura, Hideo; Tanaka, Takuo

    2008-07-01

    This study evaluated the effect of 1% hydrofluoric acid (HF) treatment on the bonding of an adhesive cement (Panavia F 2.0) to an indirect resin composite (Estenia C&B). Pairs of composite disks (10 and 8 mm in diameter by 3 mm thickness) were prepared. Adhesive surfaces were pretreated with either airborne particle abrasion or HF etching before being soaked for 30 seconds, five minutes or 10 minutes, with or without application of silane coupling agent. Adhesive specimens were fabricated by cementing a pair of treated disks. Shear bond strength was determined before and after 50,000 times of thermocycling (4 and 60 degrees C). All data were statistically analyzed using two-way ANOVA and Bonferroni's test (a=0.05). Bond strength achieved with five minutes of HF etching (18.3+/-1.1 MPa) was significantly higher (P=0.0025) than that obtained with airborne particle abrasion followed by application of silane coupling agent (14.3+/-1.8 MPa) after thermocycling.

  5. Dispersion and stability of bare hematite nanoparticles: effect of dispersion tools, nanoparticle concentration, humic acid and ionic strength.

    PubMed

    Dickson, Dionne; Liu, Guangliang; Li, Chenzhong; Tachiev, Georgio; Cai, Yong

    2012-03-01

    The aggregation and sedimentation of iron oxide nanoparticles (IONPs) can significantly affect the mobility and reactivity of IONPs and subsequently influence the interaction between IONPs and environmental contaminants. Dispersing bare IONPs into a stable suspension within nanoscale range is an important step for studying the interaction of IONPs with contaminants (e.g., toxic metals). In this study, different techniques to disperse bare IONPs (vortex, bath sonication and probe ultrasonication) and the effects of important environmental factors such as dissolved organic matter and ionic strength on the stability of IONPs dispersions were investigated. Vortex minimally dispersed IONPs with hydrodynamic diameter outside the "nano-size range" (698-2400 nm). Similar to vortex, bath sonication could not disperse IONPs efficiently. Probe ultrasonication was more effective at dispersing IONPs (50% or more) with hydrodynamic diameters ranging from 120 to 140 nm with minimal changes in size and sedimentation of IONPs for a prolonged period of time. Over the course of 168 h, considerable amounts of IONPs remained dispersed in the presence and absence of low ionic strength (0.1mM of NaCl) and 100mg/L of humic acid (HA). These results indicate that IONPs can be broken down efficiently into "nanosize range" by probe ultrasonication and a degree of stability can be achieved without the use of synthetic modifiers to enhance colloidal stability. This dispersion tool could be used to develop a laboratory method to study the adsorption mechanism between dispersed bare IONPs and toxic contaminants.

  6. Simultaneous determination of Ca, Cu, Ni, Zn and Cd binding strengths with fulvic acid fractions by Schubert's method

    USGS Publications Warehouse

    Brown, G.K.; MacCarthy, P.; Leenheer, J.A.

    1999-01-01

    The equilibrium binding of Ca2+, Ni2+, Cd2+, Cu2+ and Zn2+ with unfractionated Suwannee river fulvic acid (SRFA) and an enhanced metal binding subfraction of SRFA was measured using Schubert's ion-exchange method at pH 6.0 and at an ionic strength (??) of 0.1 (NaNO3). The fractionation and subfractionation were directed towards obtaining an isolate with an elevated metal binding capacity or binding strength as estimated by Cu2+ potentiometry (ISE). Fractions were obtained by stepwise eluting an XAD-8 column loaded with SRFA with water eluents of pH 1.0 to pH 12.0. Subfractions were obtained by loading the fraction eluted from XAD-8 at pH 5.0 onto a silica gel column and eluting with solvents of increasing polarity. Schuberts ion exchange method was rigorously tested by measuring simultaneously the conditional stability constants (K) of citric acid complexed with the five metals at pH 3.5 and 6.0. The logK of SRFA with Ca2+, Ni2+, Cd2+, Cu2+ and Zn2+ determined simultaneously at pH 6.0 follow the sequence of Cu2+>Cd2+>Ni2+>Zn2+>Ca2+ while all logK values increased for the enhanced metal binding subfraction and followed a different sequence of Cu2+>Cd2+>Ca2+>Ni2+>Zn2+. Both fulvic acid samples and citric acid exhibited a 1:1 metal to ligand stochiometry under the relatively low metal loading conditions used here. Quantitative 13C nuclear magnetic resonance spectroscopy showed increases in aromaticity and ketone content and decreases in aliphatic carbon for the elevated metal binding fraction while the carboxyl carbon, and elemental nitrogen, phosphorus, and sulfur content did not change. The more polar, elevated metal binding fraction did show a significant increase in molecular weight over the unfractionated SRFA. Copyright (C) 1999 Elsevier Science B.V.

  7. Quantitative measurement of the Brönsted acid sites in solid acids: toward a single-site design of Mo-modified ZSM-5 zeolite.

    PubMed

    Tessonnier, Jean-Philippe; Louis, Benoît; Walspurger, Stephane; Sommer, Jean; Ledoux, Marc-Jacques; Pham-Huu, Cuong

    2006-06-01

    On the basis of our previous H/D exchange studies devoted to the quantification of the number of Brönsted acid sites in solid acids, we report here an innovative approach to determine both the amount and the localization of Mo atoms inside the Mo/ZSM-5 catalyst, commonly used for the methane dehydroaromatization reaction. The influence of Mo introduction in the MFI framework was studied by means of BET, X-ray diffraction, 27Al magic angle spinning NMR, NH3 temperature-programmed desorption, and H/D isotopic exchange techniques. A dependence was found between the decrease of acidic OH groups and the Mo content. Depending on the Si/Al ratio of the zeolite, i.e., the proximity of two Brönsted acid sites, the Mo atoms substitute a different number of OH groups. Consequently, a chemical structure was proposed to describe the geometry of the Mo complex in the channels of the ZSM-5 zeolite.

  8. KDP crystal doped with L-arginine amino acid: growth, structure perfection, optical and strength characteristics

    NASA Astrophysics Data System (ADS)

    Pritula, I. M.; Kostenyukova, E. I.; Bezkrovnaya, O. N.; Kolybaeva, M. I.; Sofronov, D. S.; Dolzhenkova, E. F.; Kanaev, A.; Tsurikov, V.

    2016-07-01

    Potassium Dihydrogen Phosphate (KDP) crystal doped with L-arginine (L-arg) amino acid with 1.4 wt% concentration in the solution was grown onto a point seed by the method of temperature reduction. For the first time an attempt was made to grow large-size (7 × 6 × 8 cm3) optically transparent crystals, which allowed to analyze the effect of L-arg additive on the physical properties of the different growth sectors ({100} and {101}) of KDP. The incorporation of L-arg into both growth sectors of the crystal was confirmed by the methods of optical and IR spectroscopy and found to be caused by the ability of the amino acid to form hydrogen bonds with the face {100} and electrostatically interact with the positively charged face {101} of KDP crystal. A slight variation in the unit cell parameters was reported, the elementary cell volume of KDP:L-arg crystal increased in comparison with the one of pure KDP by 2·10-2 and 2.07·10-2 Å3 in the sectors {100} and {101}, respectively. It was found that the doping of L-arg enhanced the SHG efficiency of KDP and depended on the crystal growth sectors. The SHG efficiency of KDP:L-arg was by a factor 2.53 and 3.95 higher in comparison with those of pure KDP for {101} and {100} growth sector, respectively. The doping was found to lead to softening of both faces by ∼3-10% and ∼14-17% in the sectors {101} and {100}, respectively. Investigation of the influence of L-arg molecules on the bulk laser damage threshold of the crystals showed that the bulk laser damage threshold of the samples of KDP:L-arg crystal was higher than the one of the pure crystal in the sector {101} and lower in the sector {100}. The correlation between microhardness and laser damage threshold were discussed. The study is helpful for further searching, designing and simulation of hybrid NLO materials.

  9. Atmospheric geochemistry of formic and acetic acids at a mid-latitude temperate site

    NASA Technical Reports Server (NTRS)

    Talbot, R. W.; Beecher, K. M.; Harriss, R. C.; Cofer, R. W., III

    1988-01-01

    Tropospheric concentrations of formic and acetic acids in the gas, the aerosol, and the rainwater phases were determined in samples collected 1-2 m above ground level at an open field site in eastern Virginia. These acids were found to occur principally (98 percent or above) in the gas phase, with a marked annual seasonality, averaging 1890 ppt for formate and 1310 ppt for acetate during the growing season, as compared to 695 ppt and 700 ppt, respectively, over the nongrowing season. The data support the hypothesis that biogenic emissions from vegatation are important sources of atmospheric formic and acetic acid during the local growing season. The same time trends were observed for precipitation, although with less defined seasonality. The relative increase of the acetic acid/formic acid ratio during the nongrowing season points to the dominance of anthropogenic inputs of acetic acid from motor vehicles and biomass combustion in the wintertime.

  10. Strength and limits using 13C phospholipid fatty acid analysis in soil ecology

    NASA Astrophysics Data System (ADS)

    Watzinger, Andrea

    2016-04-01

    This presentation on microbial phospholipid biomarkers, their isotope analysis and their ability to reveal soil functions summarizes experiences gained by the author for more than 10 years. The amount and composition of phospholipid fatty acids (PLFAs) measured in environmental samples strongly depend on the methodology. To achieve comparable results the extraction, separation and methylation method must be kept constant. PLFAs patterns are sensitive to microbial community shifts even though the taxonomic resolution of PLFAs is low. The possibility to easily link lipid biomarkers with stable isotope techniques is identified as a major advantage when addressing soil functions. Measurement of PLFA isotopic ratios is sensitive and enables detecting isotopic fractionation. The difference between the carbon isotopic ratio of single PLFAs and their substrate (δ13C) can vary between -6 and +11‰. This difference derives from the fractionation during biosynthesis and from substrate inhomogeneity. Consequently, natural abundance studies are restricted to quantifying substrate uptake of the total microbial biomass. In contrast, artificial labelling enables quantifying carbon uptake into single PLFAs, but labelling success depends on homogeneous and undisturbed label application. Current developments in microbial ecology (e.g. 13C and 15N proteomics) and isotope techniques (online monitoring of CO2 isotope ratios) will likely improve soil functional interpretations in the future. 13C PLFA analysis will continue to contribute because it is affordable, sensitive and allows frequent sampling combined with the use of small amounts of 13C label.

  11. pH-adjustment strategy for volatile fatty acid production from high-strength wastewater for biological nutrient removal.

    PubMed

    Xie, Li; Liu, Hui; Chen, Yin-Guang; Zhou, Qi

    2014-01-01

    Volatile fatty acid (VFA) production from three types of high-strength organic wastewater (cassava thin stillage, starch wastewater and yellow-wine processing wastewater) were compared. The results showed that cassava thin stillage was the most suitable substrate, based on its high specific VFA production (0.68 g chemical oxygen demand (COD)/g initial soluble chemical oxygen demand (SCOD)) and yield (0.72 g COD/g SCOD) as well as low nutrient content in the substrate and fermented liquid. The acid fermented cassava thin stillage was evaluated and compared with sodium acetate in a sequencing batch reactor system. Total nitrogen removal efficiency was higher with fermented cassava thin stillage than with the sodium acetate. The effects of pH and a pH-adjustment strategy on VFA production and composition were determined using cassava thin stillage. At an initial pH range of 7-11, a relatively high VFA concentration of about 9 g COD/L was obtained. The specific VFA production (g COD/g initial SCOD) increased from 0.27 to 0.47 to 0.67 at pH 8 and from 0.26 to 0.68 to 0.81 at pH 9 (initial pH, interval pH, and constant pH adjustment, respectively). The dominant VFA species changed significantly with the increasing frequency of the pH adjustment. Further studies will examine the metabolic pathways responsible for VFA composition.

  12. A bidentate Lewis acid with a telluronium ion as an anion-binding site

    NASA Astrophysics Data System (ADS)

    Zhao, Haiyan; Gabbaï, François P.

    2010-11-01

    The search for receptors that can selectively capture small and potentially toxic anions in protic media has sparked a renewed interest in the synthesis and anion-binding properties of polydentate Lewis acids. Seeking new paradigms to enhance the anion affinities of such systems, we synthesized a bidentate Lewis acid that contains a boryl and a telluronium moiety as Lewis acidic sites. Anion-complexation studies indicate that this telluronium borane displays a high affinity for fluoride in methanol. Structural and computational studies show that the unusual fluoride affinity of this bidentate telluronium borane can be correlated with the formation of a B-F --> Te chelate motif supported by a strong lone-pair(F) --> σ*(Te-C) donor-acceptor interaction. These results, which illustrate the viability of heavier chalcogenium centres as anion-binding sites, allow us to introduce a novel strategy for the design of polydentate Lewis acids with enhanced anion affinities.

  13. Site-specific fatty acid-conjugation to prolong protein half-life in vivo.

    PubMed

    Lim, Sung In; Mizuta, Yukina; Takasu, Akinori; Hahn, Young S; Kim, Yong Hwan; Kwon, Inchan

    2013-09-10

    Therapeutic proteins are indispensable in treating numerous human diseases. However, therapeutic proteins often suffer short serum half-life. In order to extend the serum half-life, a natural albumin ligand (a fatty acid) has been conjugated to small therapeutic peptides resulting in a prolonged serum half-life via binding to patients' serum albumin in vivo. However, fatty acid-conjugation has limited applicability due to lack of site-specificity resulting in the heterogeneity of conjugated proteins and a significant loss in pharmaceutical activity. In order to address these issues, we exploited the site-specific fatty acid-conjugation to a permissive site of a protein, using copper-catalyzed alkyne-azide cycloaddition, by linking a fatty acid derivative to p-ethynylphenylalanine incorporated into a protein using an engineered pair of yeast tRNA/aminoacyl tRNA synthetase. As a proof-of-concept, we show that single palmitic acid conjugated to superfolder green fluorescent protein (sfGFP) in a site-specific manner enhanced a protein's albumin-binding in vitro about 20 times and the serum half-life in vivo 5 times when compared to those of the unmodified sfGFP. Furthermore, the fatty acid conjugation did not cause a significant reduction in the fluorescence of sfGFP. Therefore, these results clearly indicate that the site-specific fatty acid-conjugation is a very promising strategy to prolong protein serum half-life in vivo without compromising its folded structure and activity.

  14. Transcription of rat liver deoxyribonucleic acid in vitro at low ionic strength.

    PubMed Central

    Pays, E

    1978-01-01

    1. When RNA polymerase is in excess over DNA, the single-stranded breaks of DNA can be recognized as initiation sites for the ezyme. On the other hand stabel initiation complexes (resistant to inhibition by heparin) are the most abundant under these conditions. The formation of these complexes needs double-stranded DNA. It seems that RNA sequences rich in cytidine are preferentially synthesized; since rat liver DNA is A + T-rich, the transcription thus appears not to be random with respect to the base composition of DNA. 2. When the template is in excess over the polymerase, the single-stranded gaps of DNA are preferentially transcribed by rat liver RNA polymerase B and native DNA regions by Escherichia coli RNA polymerase. 3. With a large excess of DNA over the polymerase, the enzyme activity is markedly inhibited. This inhibition is proportional to the concentration of double-stranded DNA ends, but it also depends on the presence of a contaminant of DNA, removed when DNA is banded in a CsCl gradient. This contaminant could be polyphosphates. Low concentrations of spermine completely reverse this inhibition, by enhancing the rate of RNA chain elongation. 4. Double-stranded RNA is synthesized in great abundance when RNA polymerase is in excess over native DNA. Besides a majority of symmetrical sequences, stable 'hairpins' can be found. Whereas the synthesis of symmetrical sequences is more prevalent in polymerase excess, it seems that the proportion of stable 'hairpins' in RNA is independent of the polymerase/DNA ratio. PMID:32873

  15. In vitro enantioselective displacement of propranolol from protein binding sites by acetyl salicylic acid and salicylic acid.

    PubMed

    Rezaei, Z; Khabnadideh, S; Hemmateenejad, B; Dehghani, Z

    2007-09-01

    The influences of acetyl salicylic acid (ASA) and salicylic acid (SA) on the enantioselective binding of propranolol (PL) and its enantiomers to plasma proteins and human serum albumin (HSA) were investigated. The equilibrium dialysis was employed for protein binding studies. We observed statistically significant displacement of racemic-PL, (+)-(R)-PL, and (-)-(S)-PL (0.1-10 microM) from their protein binding sites by ASA (200 microg/ml) and SA (100 microg/ml). ASA and SA displaced PL stereoselectivly from its binding sites. We concluded that ASA and its metabolite SA could change R/S ratio of PL unbound fractions and they might affect pharmacokinetic properties of PL.

  16. Are carboxyl groups the most acidic sites in amino acids? Gas-phase acidities, photoelectron spectra, and computations on tyrosine, p-hydroxybenzoic acid, and their conjugate bases.

    PubMed

    Tian, Zhixin; Wang, Xue-Bin; Wang, Lai-Sheng; Kass, Steven R

    2009-01-28

    Deprotonation of tyrosine in the gas phase was found to occur preferentially at the phenolic site, and the conjugate base consists of a 70:30 mixture of phenoxide and carboxylate anions at equilibrium. This result was established by developing a chemical probe for differentiating these two isomers, and the presence of both ions was confirmed by photoelectron spectroscopy. Equilibrium acidity measurements on tyrosine indicated that deltaG(acid)(o) = 332.5 +/- 1.5 kcal mol(-1) and deltaH(acid)(o) = 340.7 +/- 1.5 kcal mol(-1). Photoelectron spectra yielded adiabatic electron detachment energies of 2.70 +/- 0.05 and 3.55 +/- 0.10 eV for the phenoxide and carboxylate anions, respectively. The H/D exchange behavior of deprotonated tyrosine was examined using three different alcohols (CF3CH2OD, C6H5CH2OD, and CH3CH2OD), and incorporation of up to three deuterium atoms was observed. Two pathways are proposed to account for these results, and all of the experimental findings are supplemented with B3LYP/aug-cc-pVDZ and G3B3 calculations. In addition, it was found that electrospray ionization of tyrosine from a 3:1 (v/v) CH3OH/H2O solution using a commercial source produces a deprotonated [M-H]- anion with the gas-phase equilibrium composition rather than the structure of the ion that exists in aqueous media. Electrospray ionization from acetonitrile, however, leads largely to the liquid-phase (carboxylate) structure. A control molecule, p-hydroxybenzoic acid, was found to behave in a similar manner. Thus, the electrospray conditions that are employed for the analysis of a compound can alter the isomeric composition of the resulting anion.

  17. Effect of strength training session on plasma amino acid concentration following oral ingestion of leucine, BCAAs or glutamine in men.

    PubMed

    Mero, Antti; Leikas, Anne; Knuutinen, Juha; Hulmi, Juha J; Kovanen, Vuokko

    2009-01-01

    We examined the acute effects of a 1-h strength training session (STS) on plasma amino acid concentration following orally ingestion of leucine, branched-chain amino acids (BCAAs) or glutamine in nine physically active men who participated in double-blinded and randomised experiments. The subjects took placebo, leucine, BCAAs, or glutamine capsules (50 mg/kg) in either rest (REST) or STS condition. Blood samples were taken before and at 30, 60, 90, and 120 min after the beginning of the treatment and they were assayed for plasma amino acids with HPLC. Following both leucine and BCAA ingestion the peak concentration of leucine was similar at rest (524 +/- 46 and 530 +/- 29 nmol/ml, respectively) and similar after STS (398 +/- 43 and 387 +/- 46 nmol/ml, respectively) but the rest and STS concentrations differed from each other (P < 0.01-0.001). The modelled polynomial data for the leucine treatment showed that the peak concentration of leucine occurred at 67 min at rest and at 90 min in STS (difference between REST and STS: P = 0.012). For the BCAA treatment the polynomial data showed that the peak concentration of leucine occurred at 72 min at rest and at 78 min in STS (P = 0.067). The peak concentration of glutamine was similar in both rest and STS condition and occurred at 60 min at rest and at 57 min in STS. In conclusion, 1-h of STS slows the increase in the peak concentration of plasma leucine similarly after oral ingestion of leucine or BCAAs but after oral ingestion of glutamine it has no slowing effect on glutamine concentration. PMID:19015870

  18. Efficient solid acid catalyst containing Lewis and Brønsted Acid sites for the production of furfurals.

    PubMed

    Mazzotta, Michael G; Gupta, Dinesh; Saha, Basudeb; Patra, Astam K; Bhaumik, Asim; Abu-Omar, Mahdi M

    2014-08-01

    Self-assembled nanoparticulates of porous sulfonated carbonaceous TiO2 material that contain Brønsted and Lewis acidic sites were prepared by a one-pot synthesis method. The material was characterized by XRD, FTIR spectroscopy, NH3 temperature-programmed desorption, pyridine FTIR spectroscopy, field-emission scanning electron microscopy, high-resolution transmission electron microscopy, N2 -sorption, atomic absorbance spectroscopy, and inductively coupled plasma optical emission spectroscopy. The carbonaceous heterogeneous catalyst (Glu-TsOH-Ti) with a Brønsted-to-Lewis acid density ratio of 1.2 and more accessible acid sites was effective to produce 5-hydroxymethylfurfural and furfural from biomass-derived mono- and disaccharides and xylose in a biphasic solvent that comprised water and biorenewable methyltetrahydrofuran. The catalyst was recycled in four consecutive cycles with a total loss of only 3 % activity. Thus, Glu-TsOH-Ti, which contains isomerization and dehydration catalytic sites and is based on a cheap and biorenewable carbon support, is a sustainable catalyst for the production of furfurals, platform chemicals for biofuels and chemicals. PMID:24807741

  19. Site-specific incorporation of a fluorescent terphenyl unnatural amino acid.

    PubMed

    Lampkowski, Jessica S; Uthappa, Diya M; Young, Douglas D

    2015-11-15

    The site-specific incorporation of unnatural amino acids into proteins has a wide range of biological implications. Of particular interest is the incorporation of fluorescent probes as a mechanism to track protein function, transport, and folding. Thus, the development of a novel system for the incorporation of new fluorescent unnatural amino acids has significant utility. Specifically, we have elucidated an aminoacyl-tRNA synthetase capable of recognizing a terphenyl UAA derivative, and charging a cognate tRNA with this amino acid for protein incorporation. Moreover, we have successfully incorporated this fluorescent UAA into GFP at several key residues, demonstrating a novel means to modulate fluorescence within the protein.

  20. Effect of acidic amino acids engineered into the active site cleft of Thermopolyspora flexuosa GH11 xylanase.

    PubMed

    Li, He; Turunen, Ossi

    2015-01-01

    Thermopolyspora flexuosa GH11 xylanase (XYN11A) shows optimal activity at pH 6-7 and 75-80 °C. We studied how mutation to aspartic acid (N46D and V48D) in the vicinity of the catalytic acid/base affects the pH activity of highly thermophilic GH11 xylanase. Both mutations shifted the pH activity profile toward acidic pH. In general, the Km values were lower at pH 4-5 than at pH 6, and in line with this, the rate of hydrolysis of xylotetraose was slightly faster at pH 4 than at pH 6. The N46D mutation and also lower pH in XYN11A increased the hydrolysis of xylotriose. The Km value increased remarkably (from 2.5 to 11.6 mg/mL) because of V48D, which indicates the weakening of binding affinity of the substrate to the active site. Xylotetraose functioned well as a substrate for other enzymes, but with lowered reaction rate for V48D. Both N46D and V48D increased the enzyme inactivation by ionic liquid [emim]OAc. In conclusion, the pH activity profile could be shifted to acidic pH due to an effect from two different directions, but the tightly packed GH11 active site can cause steric problems for the mutations.

  1. Effective and site-specific phosphoramidation reaction for universally labeling nucleic acids.

    PubMed

    Su, Yu-Chih; Chen, Hsing-Yin; Ko, Ni Chien; Hwang, Chi-Ching; Wu, Min Hui; Wang, Li-Fang; Wang, Yun-Ming; Chang, Sheng-Nan; Wang, Eng-Chi; Wang, Tzu-Pin

    2014-03-15

    Here we report efficient and selective postsynthesis labeling strategies, based on an advanced phosphoramidation reaction, for nucleic acids of either synthetic or enzyme-catalyzed origin. The reactions provided phosphorimidazolide intermediates of DNA or RNA which, whether reacted in one pot (one-step) or purified (two-step), were directly or indirectly phosphoramidated with label molecules. The acquired fluorophore-labeled nucleic acids, prepared from the phosphoramidation reactions, demonstrated labeling efficacy by their F/N ratio values (number of fluorophores per molecule of nucleic acid) of 0.02-1.2 which are comparable or better than conventional postsynthesis fluorescent labeling methods for DNA and RNA. Yet, PCR and UV melting studies of the one-step phosphoramidation-prepared FITC-labeled DNA indicated that the reaction might facilitate nonspecific hybridization in nucleic acids. Intrinsic hybridization specificity of nucleic acids was, however, conserved in the two-step phosphoramidation reaction. The reaction of site-specific labeling nucleic acids at the 5'-end was supported by fluorescence quenching and UV melting studies of fluorophore-labeled DNA. The two-step phosphoramidation-based, effective, and site-specific labeling method has the potential to expedite critical research including visualization, quantification, structural determination, localization, and distribution of nucleic acids in vivo and in vitro.

  2. In Situ Oxalic Acid Injection to Accelerate Arsenic Remediation at a Superfund Site in New Jersey

    PubMed Central

    Wovkulich, Karen; Stute, Martin; Mailloux, Brian J.; Keimowitz, Alison R.; Ross, James; Bostick, Benjamin; Sun, Jing; Chillrud, Steven N.

    2015-01-01

    Arsenic is a prevalent contaminant at a large number of US Superfund sites; establishing techniques that accelerate As remediation could benefit many sites. Hundreds of tons of As were released into the environment by the Vineland Chemical Co. in southern New Jersey during its manufacturing lifetime (1949–1994), resulting in extensive contamination of surface and subsurface soils and sediments, groundwater, and the downstream watershed. Despite substantial intervention at this Superfund site, sufficient aquifer cleanup could require many decades if based on traditional pump and treat technologies only. Laboratory column experiments have suggested that oxalic acid addition to contaminated aquifer solids could promote significant As release from the solid phase. To evaluate the potential of chemical additions to increase As release in situ and boost treatment efficiency, a forced gradient pilot scale study was conducted on the Vineland site. During spring/summer 2009, oxalic acid and bromide tracer were injected into a small portion (~50 m2) of the site for 3 months. Groundwater samples indicate that introduction of oxalic acid led to increased As release. Between 2.9 and 3.6 kg of As were removed from the sampled wells as a result of the oxalic acid treatment during the 3-month injection. A comparison of As concentrations on sediment cores collected before and after treatment and analyzed using X-ray fluorescence spectroscopy suggested reduction in As concentrations of ~36% (median difference) to 48% (mean difference). While further study is necessary, the addition of oxalic acid shows potential for accelerating treatment of a highly contaminated site and decreasing the As remediation time-scale. PMID:25598701

  3. Absolute acidity of clay edge sites from ab-initio simulations

    NASA Astrophysics Data System (ADS)

    Tazi, Sami; Rotenberg, Benjamin; Salanne, Mathieu; Sprik, Michiel; Sulpizi, Marialore

    2012-10-01

    We provide a microscopic understanding of the solvation structure and reactivity of the edges of neutral clays. In particular we address the tendency to deprotonation of the different reactive groups on the (0 1 0) face of pyrophyllite. Such information cannot be inferred directly from titration experiments, which do not discriminate between different sites and whose interpretation resorts to macroscopic models. The determination of the corresponding pKa then usually relies on bond valence models, sometimes improved by incorporating some structural information from ab-initio simulations. Here we use density functional theory based molecular dynamics simulations, combined with thermodynamic integration, to compute the free energy of the reactions of water with the different surface groups, leading to a deprotonated site and an aqueous hydronium ion. Our approach consistently describes the clay and water sides of the interface and includes naturally electronic polarization effects. It also allows to investigate the structure and solvation of all sites separately. We find that the most acidic group is SiOH, due to its ability to establish strong hydrogen bonds with adsorbed water, as it also happens on the quartz and amorphous silica surfaces. The acidity constant of AlOH2 is only 1 pKa unit larger. Finally, the pKa of AlOH is outside the possible range in water and this site should not deprotonate in aqueous solution. We show that the solvation of surface sites and hence their acidity is strongly affected by the proximity of other sites, in particular for AlOH and AlOH2 which share the same Al. We discuss the implications of our findings on the applicability of bond valence models to predict the acidity of edge sites of clays.

  4. PHYSICAL SOLUTIONS FOR ACID ROCK DRAINAGE AT REMOTE SITES DEMONSTRATION PROJECT

    EPA Science Inventory

    This report summarizes the results of Mine Waste Technology Program, Activity III, Project 42, Physical Solutions for Acid Rock Drainage at Remote Sites, funded by the U.S. Environmental Protection Agency (EPA) and jointly administered by EPA and the U.S. Department of Energy. A...

  5. LIME TREATMENT LAGOONS TECHNOLOGY FOR TREATING ACID MINE DRAINAGE FROM TWO MINING SITES

    EPA Science Inventory

    Runoff and drainage from active and inactive mines are someof the most environmentally damaging land uses i the US. Acid Mine drainage (AMD) from mining sites across the country requires treatment because of high metal concentrations that exceed regulatory standards for safe disc...

  6. Modeling Analysis for Characterizing Sulfate Reduction at an Acid Mine Drainage Site

    NASA Astrophysics Data System (ADS)

    Sengupta, A.; Ahlfeld, D. P.

    2004-05-01

    A field site has been established at Davis Mine, an abandoned pyrite mine in rural Rowe, Massachusetts in the United States. At the site, attenuation restricts the extent of AMD in both the groundwater and surface water of the area. Current research is examining the Fe(III) and sulfate reduction along with a complex community of acidophilic and acid-tolerant anaerobic microorganisms. In an effort to interlink the geochemical reduction with the microbial community existing in the site, the role of the Fe(III) and sulfate reducing bacteria is being investigated. Initial experimental data and column studies have shown the presence of sulfate reducing bacteria at the site. A detailed groundwater flow model for the affected site has been developed. A model is currently being developed of the various geochemical and biological processes at Davis Mine for use in distinguishing between sulfate reduction and dilution as they affect observed sulfate attenuation.

  7. RF1 Knockout Allows Ribosomal Incorporation of Unnatural Amino Acids at Multiple Sites

    PubMed Central

    Johnson, David B.F.; Xu, Jianfeng; Shen, Zhouxin; Takimoto, Jeffrey K.; Schultz, Matthew D.; Schmitz, Robert J.; Ecker, Joseph R.; Briggs, Steven P.; Wang, Lei

    2011-01-01

    Stop codons have been exploited for genetic incorporation of unnatural amino acids (Uaas) in live cells, but the efficiency is low possibly due to competition from release factors, limiting the power and scope of this technology. Here we show that the reportedly essential release factor 1 can be knocked out from Escherichia coli by fixing release factor 2. The resultant strain JX33 is stable and independent, and reassigns UAG from a stop signal to an amino acid when a UAG-decoding tRNA/synthetase pair is introduced. Uaas were efficiently incorporated at multiple UAG sites in the same gene without translational termination in JX33. We also found that amino acid incorporation at endogenous UAG codons is dependent on RF1 and mRNA context, which explains why E. coli tolerates apparent global suppression of UAG. JX33 affords a unique autonomous host for synthesizing and evolving novel protein functions by enabling Uaa incorporation at multiple sites. PMID:21926996

  8. The Optimal Suture Site for the Repair of Posterior Horn Root Tears: Biomechanical Evaluation of Pullout Strength in Porcine Menisci

    PubMed Central

    Kim, Young-Mo; Noh, Chang-Kyun; Park, Il-Young

    2016-01-01

    Purpose There are few studies on biomechanical evaluation of suture points in repair of root tears. The purpose of this study was to determine the point of greatest pullout strength for root tear repair. Materials and Methods A total of 120 fresh porcine medial menisci were obtained. The red-red and red-white zones of the meniscus were divided by two lines designated as lines A and B (groups A and B). Groups A and B were further divided into three groups each by dividing lines A and B into three points: 3, 5, and 7 mm from the meniscal ligament root insertion. Vertical meniscal repair was performed on each point. The pullout failure strength was tested using a biaxial servohydraulic testing machine. Results The average maximal load at failure was significantly greater in group A than group B (87.65 vs. 62.93; p<0.001) The average length at maximal load failure was greater in group A than group B (4.35 vs. 3.2; p<0.001). Among the subgroups of 3, 5, and 7 mm in both groups A and B, 7 mm showed the greatest maximal load (p<0.001). Conclusions The pullout strength was statistically significantly greater in group A than group B and in the 7 mm subgroup than the 3 and 5 mm subgroups. Thus, the 7 mm subgroup in group A showed the greatest pullout strength. PMID:27274472

  9. Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

    SciTech Connect

    Gangi Setty, Thanuja; Cho, Christine; Govindappa, Sowmya; Apicella, Michael A.; Ramaswamy, S.

    2014-07-01

    Structure–function studies of sialic acid-binding proteins from F. nucleatum, P. multocida, V. cholerae and H. influenzae reveal a conserved network of hydrogen bonds involved in conformational change on ligand binding. Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states.

  10. Transport and Retention of TiO2 Rutile Nanoparticles in Saturated Porous Media: Influence of Solution pH, Ionic Strength, and the Presence of Humic Acid

    EPA Science Inventory

    The influence of solution pH, ionic strength, and varying concentrations of the Suwannee River Humic Acid (SRHA) on the transport of titanium dioxide (TiO2, rutile) nanoparticle aggregates (nTiO2) in saturated porous media was investigated through systematically examining the tra...

  11. Is There a Need to Discuss Atomic Orbital Overlap When Teaching Hydrogen-Halide Bond Strength and Acidity Trends in Organic Chemistry?

    ERIC Educational Resources Information Center

    Devarajan, Deepa; Gustafson, Samantha J.; Bickelhaupt, F. Matthias; Ess, Daniel H.

    2015-01-01

    Undergraduate organic chemistry textbooks and Internet websites use a variety of approaches for presenting and explaining the impact of halogen atom size on trends in bond strengths and/or acidity of hydrogen halides. In particular, several textbooks and Internet websites explain these trends by invoking decreasing orbital overlap between the…

  12. Adsorption of Carbon Dioxide on Unsaturated Metal Sites in M2 (dobpdc) Frameworks with Exceptional Structural Stability and Relation between Lewis Acidity and Adsorption Enthalpy.

    PubMed

    Yoo, Ga Young; Lee, Woo Ram; Jo, Hyuna; Park, Joonho; Song, Jeong Hwa; Lim, Kwang Soo; Moon, Dohyun; Jung, Hyun; Lim, Juhyung; Han, Sang Soo; Jung, Yousung; Hong, Chang Seop

    2016-05-23

    A series of metal-organic frameworks (MOFs) M2 (dobpdc) (M=Mn, Co, Ni, Zn; H4 dobpdc=4,4'-dihydroxy-1,1'-biphenyl-3,3'-dicarboxylic acid), with a highly dense arrangement of open metal sites along hexagonal channels were prepared by microwave-assisted or simple solvothermal reactions. The activated materials were structurally expanded when guest molecules including CO2 were introduced into the pores. The Lewis acidity of the open metal sites varied in the order MnZn, as confirmed by C=O stretching bands in the IR spectra, which are related to the CO2 adsorption enthalpy. DFT calculations revealed that the high CO2 binding affinity of transition-metal-based M2 (dobpdc) is primarily attributable to the favorable charge transfer from CO2 (oxygen lone pair acting as a Lewis base) to the open metal sites (Lewis acid), while electrostatic effects, the underlying factor responsible for the particular order of binding strength observed across different transition metals, also play a role. The framework stability against water coincides with the order of Lewis acidity. In this series of MOFs, the structural stability of Ni2 (dobpdc) is exceptional; it endured in water vapor, liquid water, and in refluxing water for one month, and the solid remained intact on exposure to solutions of pH 2-13. The DFT calculations also support the experimental finding that Ni2 (dobpdc) has higher chemical stability than the other frameworks.

  13. Adsorption of Carbon Dioxide on Unsaturated Metal Sites in M2 (dobpdc) Frameworks with Exceptional Structural Stability and Relation between Lewis Acidity and Adsorption Enthalpy.

    PubMed

    Yoo, Ga Young; Lee, Woo Ram; Jo, Hyuna; Park, Joonho; Song, Jeong Hwa; Lim, Kwang Soo; Moon, Dohyun; Jung, Hyun; Lim, Juhyung; Han, Sang Soo; Jung, Yousung; Hong, Chang Seop

    2016-05-23

    A series of metal-organic frameworks (MOFs) M2 (dobpdc) (M=Mn, Co, Ni, Zn; H4 dobpdc=4,4'-dihydroxy-1,1'-biphenyl-3,3'-dicarboxylic acid), with a highly dense arrangement of open metal sites along hexagonal channels were prepared by microwave-assisted or simple solvothermal reactions. The activated materials were structurally expanded when guest molecules including CO2 were introduced into the pores. The Lewis acidity of the open metal sites varied in the order MnZn, as confirmed by C=O stretching bands in the IR spectra, which are related to the CO2 adsorption enthalpy. DFT calculations revealed that the high CO2 binding affinity of transition-metal-based M2 (dobpdc) is primarily attributable to the favorable charge transfer from CO2 (oxygen lone pair acting as a Lewis base) to the open metal sites (Lewis acid), while electrostatic effects, the underlying factor responsible for the particular order of binding strength observed across different transition metals, also play a role. The framework stability against water coincides with the order of Lewis acidity. In this series of MOFs, the structural stability of Ni2 (dobpdc) is exceptional; it endured in water vapor, liquid water, and in refluxing water for one month, and the solid remained intact on exposure to solutions of pH 2-13. The DFT calculations also support the experimental finding that Ni2 (dobpdc) has higher chemical stability than the other frameworks. PMID:27105924

  14. The influence of chemisorbed molecules on mass transfer in H-ZSM-5-type zeolites and the location of Broensted acid sites

    SciTech Connect

    Caro, J.; Buelow, M. ); Kaerger, J.; Pfeifer, H. )

    1988-11-01

    Heterogeneous catalysis is one of the most important applications of zeolites. Therefore, various methods have been developed to determine the strength and concentration of Bronsted acid sites in zeolites. Among them, in the last few years, {sup 1}H MAS NMR has become a powerful tool. In addition to the accessibility of the acid sites probed by chemisorption of N-bases, the steric environment of these catalytically active sites is of importance since it imposes constraints on the geometry of the transition state. However, only a few studies have been reported on this topic. Information was obtained from quantum chemical calculations, catalytic experiments, I.R. spectroscopy, and the arrangement of guest molecules. From these investigations it has been concluded that in H-ZSM-5 the channel intersections should be preferential location centers for the Bronsted acid sites. In adsorption technology, in the use of zeolites as shape-selective adsorbents, modification of the molecular sieve properties by chemisorption of nitrogen-containing bases (N-compounds) has become a common technique. The authors have applied the NMR pulsed field gradient technique to study the influence of chemisorbed N-compounds on transport properties of molecular sieves, considering the chemisorbed compounds as transport obstacles.

  15. Lipid Classes and Fatty Acids in Ophryotrocha cyclops, a Dorvilleid from Newfoundland Aquaculture Sites

    PubMed Central

    Salvo, Flora; Dufour, Suzanne C.; Hamoutene, Dounia; Parrish, Christopher C.

    2015-01-01

    A new opportunistic annelid (Ophryotrocha cyclops) discovered on benthic substrates underneath finfish aquaculture sites in Newfoundland (NL) may be involved in the remediation of organic wastes. At those aquaculture sites, bacterial mats and O. cyclops often coexist and are used as indicators of organic enrichment. Little is known on the trophic strategies used by these annelids, including whether they might consume bacteria or other aquaculture-derived wastes. We studied the lipid and fatty acid composition of the annelids and their potential food sources (degraded flocculent organic matter, fresh fish pellets and bacterial mats) to investigate feeding relationships in these habitats and compared the lipid and fatty acid composition of annelids before and after starvation. Fish pellets were rich in lipids, mainly terrestrially derived C18 fatty acids (18:1ω9, 18:2ω6, 18:3ω3), while bacterial samples were mainly composed of ω7 fatty acids, and flocculent matter appeared to be a mixture of fresh and degrading fish pellets, feces and bacteria. Ophryotrocha cyclops did not appear to store excessive amounts of lipids (13%) but showed a high concentration of ω3 and ω6 fatty acids, as well as a high proportion of the main fatty acids contained in fresh fish pellets and bacterial mats. The dorvilleids and all potential food sources differed significantly in their lipid and fatty acid composition. Interestingly, while all food sources contained low proportions of 20:5ω3 and 20:2ω6, the annelids showed high concentrations of these two fatty acids, along with 20:4ω6. A starvation period of 13 days did not result in a major decrease in total lipid content; however, microscopic observations revealed that very few visible lipid droplets remained in the gut epithelium after three months of starvation. Ophryotrocha cyclops appears well adapted to extreme environments and may rely on lipid-rich organic matter for survival and dispersal in cold environments. PMID:26308719

  16. Lipid Classes and Fatty Acids in Ophryotrocha cyclops, a Dorvilleid from Newfoundland Aquaculture Sites.

    PubMed

    Salvo, Flora; Dufour, Suzanne C; Hamoutene, Dounia; Parrish, Christopher C

    2015-01-01

    A new opportunistic annelid (Ophryotrocha cyclops) discovered on benthic substrates underneath finfish aquaculture sites in Newfoundland (NL) may be involved in the remediation of organic wastes. At those aquaculture sites, bacterial mats and O. cyclops often coexist and are used as indicators of organic enrichment. Little is known on the trophic strategies used by these annelids, including whether they might consume bacteria or other aquaculture-derived wastes. We studied the lipid and fatty acid composition of the annelids and their potential food sources (degraded flocculent organic matter, fresh fish pellets and bacterial mats) to investigate feeding relationships in these habitats and compared the lipid and fatty acid composition of annelids before and after starvation. Fish pellets were rich in lipids, mainly terrestrially derived C18 fatty acids (18:1ω9, 18:2ω6, 18:3ω3), while bacterial samples were mainly composed of ω7 fatty acids, and flocculent matter appeared to be a mixture of fresh and degrading fish pellets, feces and bacteria. Ophryotrocha cyclops did not appear to store excessive amounts of lipids (13%) but showed a high concentration of ω3 and ω6 fatty acids, as well as a high proportion of the main fatty acids contained in fresh fish pellets and bacterial mats. The dorvilleids and all potential food sources differed significantly in their lipid and fatty acid composition. Interestingly, while all food sources contained low proportions of 20:5ω3 and 20:2ω6, the annelids showed high concentrations of these two fatty acids, along with 20:4ω6. A starvation period of 13 days did not result in a major decrease in total lipid content; however, microscopic observations revealed that very few visible lipid droplets remained in the gut epithelium after three months of starvation. Ophryotrocha cyclops appears well adapted to extreme environments and may rely on lipid-rich organic matter for survival and dispersal in cold environments.

  17. Characterization of Naphthaleneacetic Acid Binding to Receptor Sites on Cellular Membranes of Maize Coleoptile Tissue 1

    PubMed Central

    Ray, Peter M.; Dohrmann, Ulrike; Hertel, Rainer

    1977-01-01

    Characteristics of and optimum conditions for saturable (“specific”) binding of [14C]naphthaleneacetic acid to sites located on membranous particles from maize (Zea mays L.) coleoptiles are described. Most, if not all, of the specific binding appears to be due to a single kinetic class of binding sites having a KD of 5 to 7 × 10−7m for naphthalene-1-acetic acid (NAA). Binding of NAA is insensitive to high monovalent salt concentrations, indicating that binding is not primarily ionic. However, specific binding is inhibited by Mg2+ or Ca2+ above 5 mm. Specific binding is improved by organic acids, especially citrate. Binding is heat-labile and is sensitive to agents that act either on proteins or on lipids. Specific binding is reversibly inactivated by reducing agents such as dithioerythritol; a reducible group, possibly a disulfide group, may be located at the binding site and required for its function. The affinity of the specific binding sites for auxins is modified by an unidentified dialyzable, heat-stable, apparently amphoteric, organic factor (“supernatant factor”) found in maize tissue. PMID:16659851

  18. Effects of Short-Term Docosahexaenoic Acid Supplementation on Markers of Inflammation after Eccentric Strength Exercise in Women

    PubMed Central

    Corder, Katherine E.; Newsham, Katherine R.; McDaniel, Jennifer L.; Ezekiel, Uthayashanker R.; Weiss, Edward P.

    2016-01-01

    The omega-3 fatty acid docosahexaenoic acid (DHA) has anti-inflammatory and anti-nociceptive (pain inhibiting) effects. Because strenuous exercise often results in local inflammation and pain, we hypothesized that DHA supplementation attenuates the rise in markers of local muscle inflammation and delayed onset muscle soreness (DOMS) that occur after eccentric strength exercise. Twenty-seven, healthy women (33 ± 2 y, BMI 23.1±1.0 kg·m-2) were randomized to receive 9d of 3000 mg/d DHA or placebo in a double-blind fashion. On day 7 of the supplementation period, the participants performed 4 sets of maximal-effort eccentric biceps curl exercise. Before and 48h after the eccentric exercise, markers of inflammation were measured including measures of muscle soreness (10-point visual analog pain scale, VAS), swelling (arm circumference), muscle stiffness (active and passive elbow extension), skin temperature, and salivary C-reactive protein (CRP) concentrations. As expected, muscle soreness and arm circumference increased while active and passive elbow extension decreased. The increase in soreness was 23% less in the DHA group (48h increase in VAS soreness ratings: 4.380.4 vs. 5.600.5, p=0.02). Furthermore, the number of subjects who were able to achieve full active elbow extension 48h after eccentric exercise was greater in the DHA group (71% vs. 15%, p = 0.006), indicating significantly less muscle stiffness. No between-group differences were observed for passive elbow extension (p = 0.78) or arm swelling (p = 0.75). Skin temperature and salivary CRP concentrations did not change from baseline to 48h after exercise in either group. These findings indicate that short-term DHA supplementation reduces exercise-induced muscle soreness and stiffness. Therefore, in addition to other health benefits that n-3 fatty acids have been associated with, DHA supplementation could be beneficial for improving tolerance to new and/or strenuous exercise programs and thereby might

  19. Zinc-induced oligomerization of zinc α2 glycoprotein reveals multiple fatty acid-binding sites.

    PubMed

    Zahid, Henna; Miah, Layeque; Lau, Andy M; Brochard, Lea; Hati, Debolina; Bui, Tam T T; Drake, Alex F; Gor, Jayesh; Perkins, Stephen J; McDermott, Lindsay C

    2016-01-01

    Zinc α2 glycoprotein (ZAG) is an adipokine with a class I MHC protein fold and is associated with obesity and diabetes. Although its intrinsic ligand remains unknown, ZAG binds the dansylated C11 fatty acid 11-(dansylamino)undecanoic acid (DAUDA) in the groove between the α1 and α2 domains. The surface of ZAG has approximately 15 weak zinc-binding sites deemed responsible for precipitation from human plasma. In the present study the functional significance of these metal sites was investigated. Analytical ultracentrifugation (AUC) and CD showed that zinc, but not other divalent metals, causes ZAG to oligomerize in solution. Thus ZAG dimers and trimers were observed in the presence of 1 and 2 mM zinc. Molecular modelling of X-ray scattering curves and sedimentation coefficients indicated a progressive stacking of ZAG monomers, suggesting that the ZAG groove may be occluded in these. Using fluorescence-detected sedimentation velocity, these ZAG-zinc oligomers were again observed in the presence of the fluorescent boron dipyrromethene fatty acid C16-BODIPY (4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-hexadecanoic acid). Fluorescence spectroscopy confirmed that ZAG binds C16-BODIPY. ZAG binding to C16-BODIPY, but not to DAUDA, was reduced by increased zinc concentrations. We conclude that the lipid-binding groove in ZAG contains at least two distinct fatty acid-binding sites for DAUDA and C16-BODIPY, similar to the multiple lipid binding seen in the structurally related immune protein CD1c. In addition, because high concentrations of zinc occur in the pancreas, the perturbation of these multiple lipid-binding sites by zinc may be significant in Type 2 diabetes where dysregulation of ZAG and zinc homoeostasis occurs.

  20. Evidence of sulphur and nitrogen deposition signals at the United Kingdom Acid Waters Monitoring Network sites.

    PubMed

    Cooper, D M

    2005-09-01

    Some recent studies of trends in sulphate in surface waters have alluded to possible lag effects imposed by catchment soils, resulting in discrepancies between trends in deposition and run-off. To assess the extent of these possible effects in the UK, sulphate concentration data from the United Kingdom Acid Waters Monitoring Network (AWMN) sites are compared with estimates of sulphur deposition at each site. From these data, input-output budgets are computed at an annual time scale. The estimated budgets suggest a close association between catchment sulphur inputs and outputs at an annual scale, with well-balanced annual budgets at most sites, indicative of only minor lag effects. A similar analysis of the AWMN site nitrogen budget shows little evidence of an association between nitrogen inputs and outputs at this time scale.

  1. Modeling protein evolution with several amino acid replacement matrices depending on site rates.

    PubMed

    Le, Si Quang; Dang, Cuong Cao; Gascuel, Olivier

    2012-10-01

    Most protein substitution models use a single amino acid replacement matrix summarizing the biochemical properties of amino acids. However, site evolution is highly heterogeneous and depends on many factors that influence the substitution patterns. In this paper, we investigate the use of different substitution matrices for different site evolutionary rates. Indeed, the variability of evolutionary rates corresponds to one of the most apparent heterogeneity factors among sites, and there is no reason to assume that the substitution patterns remain identical regardless of the evolutionary rate. We first introduce LG4M, which is composed of four matrices, each corresponding to one discrete gamma rate category (of four). These matrices differ in their amino acid equilibrium distributions and in their exchangeabilities, contrary to the standard gamma model where only the global rate differs from one category to another. Next, we present LG4X, which also uses four different matrices, but leaves aside the gamma distribution and follows a distribution-free scheme for the site rates. All these matrices are estimated from a very large alignment database, and our two models are tested using a large sample of independent alignments. Detailed analysis of resulting matrices and models shows the complexity of amino acid substitutions and the advantage of flexible models such as LG4M and LG4X. Both significantly outperform single-matrix models, providing gains of dozens to hundreds of log-likelihood units for most data sets. LG4X obtains substantial gains compared with LG4M, thanks to its distribution-free scheme for site rates. Since LG4M and LG4X display such advantages but require the same memory space and have comparable running times to standard models, we believe that LG4M and LG4X are relevant alternatives to single replacement matrices. Our models, data, and software are available from http://www.atgc-montpellier.fr/models/lg4x.

  2. An improved thermodynamic model for the complexation of trivalent actinides and lanthanide with oxalic acid valid to high ionic strength.

    DOE PAGES

    Xiong, Yongliang; Thakur, Punam; Borkowski, Marian

    2015-07-30

    The dissociation constants of oxalic acid (Ox), and the stability constants of Am3+, Cm3+ and Eu3+ with Ox2– have been determined at 25 °C, over a range of concentration varying from 0.1 to 6.60 m NaClO4 using potentiometric titration and extraction techniques, respectively. The experimental data support the formation of complexes, M(Ox)n3 – 2n, where (M = Am3+, Cm3+ and Eu3+ and n = 1 and 2). The dissociation constant and the stability constant values measured as a function of NaClO4 concentration were used to estimate the Pitzer parameters for the respective interactions of Am3+, Cm3+ and Eu3+ with Ox.more » Furthermore, the stability constants data of Am3+ –Ox measured in NaClO4 and in NaCl solutions from the literature were simultaneously fitted in order to refine the existing actinide–oxalate complexation model that can be used universally in the safety assessment of radioactive waste disposal. The thermodynamic stability constant: log β0101 = 6.30 ± 0.06 and log β0102 = 10.84 ± 0.06 for Am3+ was obtained by simultaneously fitting data in NaCl and NaClO4 media. Additionally, log β0101 = 6.72 ± 0.08 and log β0102 = 11.05 ± 0.09 for the Cm3+ and log β0101 = 6.67 ± 0.08 and log β0102 = 11.15 ± 0.09 for the Eu3+ were calculated by extrapolation of data to zero ionic strength in NaClO4 medium only. For all stability constants, the Pitzer model gives an excellent representation of the data using interaction parameters β(0), β(1), and CΦ determined in this work. The thermodynamic model developed in this work will be useful in accurately modeling the potential solubility of trivalent actinides and early lanthanides to ionic strength of 6.60 m in low temperature environments in the presence of Ox. Furthermore, the work is also applicable to the accurate modeling transport of rare earth elements in various environments under the surface conditions.« less

  3. Shear Bond Strength of an Etch-and-rinse Adhesive to Er:YAG Laser- and/or Phosphoric Acid-treated Dentin

    PubMed Central

    Davari, Abdolrahim; Sadeghi, Mostafa; Bakhshi, Hamid

    2013-01-01

    Background and aims. Er:YAG laser irradiation has been claimed to improve the adhesive properties of dentin; therefore, it has been proposed as an alternative to acid etching. The aim of this in vitro study was to investigate the shear bond strength of an etch-and-rinse adhesive system to dentin surfaces following Er:YAG laser and/or phosphoric acid etching. Materials and methods. The roots of 75 sound maxillary premolars were sectioned below the CEJ and the crowns were embedded in auto-polymerizing acrylic resin with the buccal surfaces facing up. The buccal surfaces were ground using a diamond bur and polished until the dentin was exposed; the samples were randomly divided into five groups (n=15) according to the surface treatment: (1) acid etching; (2) laser etching; (3) laser etching followed by acid etching; (4) acid etching followed by laser etching and (5) no acid etching and no laser etching (control group). Composite resin rods (Point 4, Kerr Co) were bonded to treated dentin surfaces with an etch-and-rise adhesive system (Optibond FL, Kerr Co) and light-cured.After storage for two weeks at 37°C and 100% humidity and then thermocycling, bond strength was measured with a Zwick Universal Testing Machine at a crosshead speed of 1 mm/min. Data was analyzed using parametric and non-parametric tests (P<0.05). Results. Mean shear bond strength for acid etching (20.1±1.8 MPa) and acid+laser (15.6±3.5 MPa) groups were significantly higher than those for laser+acid (15.6±3.5 MPa), laser etching (14.1±3.4 MPa) and control (8.1±2.1 MPa) groups. However, there were no significant differences between acid etching and acid+laser groups, and between laser+acid and laser groups. Conclusion. When the cavity is prepared by bur, it is not necessary to etch the dentin surface by Er:YAG laser following acid etching and acid etching after laser etching. PMID:23875083

  4. Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

    PubMed Central

    Gangi Setty, Thanuja; Cho, Christine; Govindappa, Sowmya; Apicella, Michael A.; Ramaswamy, S.

    2014-01-01

    Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states. PMID:25004958

  5. Mutation-selection models of coding sequence evolution with site-heterogeneous amino acid fitness profiles.

    PubMed

    Rodrigue, Nicolas; Philippe, Hervé; Lartillot, Nicolas

    2010-03-01

    Modeling the interplay between mutation and selection at the molecular level is key to evolutionary studies. To this end, codon-based evolutionary models have been proposed as pertinent means of studying long-range evolutionary patterns and are widely used. However, these approaches have not yet consolidated results from amino acid level phylogenetic studies showing that selection acting on proteins displays strong site-specific effects, which translate into heterogeneous amino acid propensities across the columns of alignments; related codon-level studies have instead focused on either modeling a single selective context for all codon columns, or a separate selective context for each codon column, with the former strategy deemed too simplistic and the latter deemed overparameterized. Here, we integrate recent developments in nonparametric statistical approaches to propose a probabilistic model that accounts for the heterogeneity of amino acid fitness profiles across the coding positions of a gene. We apply the model to a dozen real protein-coding gene alignments and find it to produce biologically plausible inferences, for instance, as pertaining to site-specific amino acid constraints, as well as distributions of scaled selection coefficients. In their account of mutational features as well as the heterogeneous regimes of selection at the amino acid level, the modeling approaches studied here can form a backdrop for several extensions, accounting for other selective features, for variable population size, or for subtleties of mutational features, all with parameterizations couched within population-genetic theory. PMID:20176949

  6. Site-specific study on stabilization of acid-generating mine tailings using coal fly ash

    SciTech Connect

    Shang, J.Q.; Wang, H.L.; Kovac, V.; Fyfe, J.

    2006-03-15

    A site-specific study on stabilizing acid-generating mine tailings from Sudbury Mine using a coal fly ash from Nanticoke Generating Station is presented in this paper. The objective of the study is to evaluate the feasibility of codisposal of the fly ash and mine tailings to reduce environmental impacts of Sudbury tailings disposal sites. The study includes three phases, i.e., characterization of the mine tailings, and coal fly ash, oxidation tests on the mine tailings and kinetic column permeation tests. The results of the experiments indicate that when permeated with acid mine drainage, the hydraulic conductivity of Nanticoke coal fly ash decreased more than three orders of magnitude (from 1 x 10{sup -6} to 1 x 10{sup -9} cm/s), mainly due to chemical reactions between the ash solids and acid mine drainage. Furthermore, the hydraulic gradient required for acid mine drainage to break through the coal fly ash is increased up to ten times (from 17 to 150) as compared with that for water. The results also show that the leachate from coal fly ash neutralizes the acidic pore fluid of mine tailings. The concentrations of trace elements in effluents from all kinetic column permeation tests indicated that coplacement of coal fly ash with mine tailings has the benefit of immobilizing trace elements, especially heavy metals. All regulated element concentrations from effluent during testing are well below the leachate quality criteria set by the local regulatory authority.

  7. The control of acid mine drainage at the Summitville Mine Superfund Site

    SciTech Connect

    Ketellapper, V.L.; Williams, L.O.

    1996-11-01

    The Summitville Mine Superfund Site is located about 25 miles south of Del Norte, Colorado, in Rio Grande County. Occurring at an average elevation of 11,500 feet in the San Juan Mountain Range, the mine site is located two miles east of the Continental Divide. Mining at Summitville has occurred since 1870. The mine was most recently operated by Summitville Consolidated Mining Company, Inc. (SCMCI) as an open pit gold mine with extraction by means of a cyanide leaching process. In December of 1992, SCMCI declared bankruptcy and vacated the mine site. At that time, the US Environmental Protection Agency (EPA) took over operations of the water treatment facilities to prevent a catastrophic release of cyanide and metal-laden water from the mine site. Due to high operational costs of water treatment (approximately $50,000 per day), EPA established a goal to minimize active water treatment by reducing or eliminating acid mine drainage (AMD). All of the sources of AMD generation on the mine site were evaluated and prioritized. Of the twelve areas identified as sources of AMD, the Cropsy Waste Pile, the Summitville Dam Impoundment, the Beaver Mud Dump, the Reynolds and Chandler adits, and the Mine Pits were consider to be the most significant contributors to the generation of metal-laden acidic (low pH) water. A two part plan was developed to control AMD from the most significant sources. The first part was initiated immediately to control AMD being released from the Site. This part focused on improving the efficiency of the water treatment facilities and controlling the AMD discharges from the mine drainage adits. The second part of the plan was aimed at reducing the AMD generated in groundwater and surface water runoff from the mine wastes. A lined and capped repository located in the mine pits for acid generating mining waste and water treatment plant sludge was found to be the most feasible alternative.

  8. Moleculary imprinted polymers with metalloporphyrin-based molecular recognition sites coassembled with methacrylic acid.

    PubMed

    Takeuchi, T; Mukawa, T; Matsui, J; Higashi, M; Shimizu, K D

    2001-08-15

    A diastereoselective molecularly imprinted polymer (MIP) for (-)-cinchonidine, PPM(CD), was prepared by the combined use of methacrylic acid and vinyl-substituted zinc(II) porphyrin as functional monomers. Compared to MIPs using only methacrylic acid or zinc porphyrin as a functional monomer, PM(CD) and PP(CD), respectively, PPM(CD) showed higher binding ability for (-)-cinchonidine in chromatographic tests using the MIP-packed columns. Scatchard analysis gave a higher association constant of PPM(CD) for (-)-cinchonidine (1.14 x 10(7) M(-1)) than those of PP(CD) (1.45 x 10(6) M(-1)) and PM(CD) (6.78 x 10(6) M(-1)). The affinity distribution of binding sites estimated by affinity spectrum analysis showed a higher percentage of high-affinity sites and a lower percentage of low-affinity sites in PPM(CD). The MIPs containing a zinc(II) porphyrin in the binding sites, PPM(CD) and PP(CD), showed fluorescence quenching according to the binding of (-)-cinchonidine, and the quenching was significant in the low-concentration range, suggesting that the high-affinity binding sites contain the porphyrin residue. The correlation of the relative fluorescence intensity against log of (-)-cinchonidine concentrations showed a linear relationship. These results revealed that the MIP having highly specific binding sites was assembled by the two functional monomers, vinyl-substituted zinc(II) porphyrin and methacrylic acid, and they cooperatively worked to yield the specific binding. In addition, the zinc(II) porphyrin-based MIPs appeared to act as fluorescence sensor selectively responded by binding events of the template molecule.

  9. Effect of experimental acid/base conditioner on microtensile bond strength of 4-META/MMA-TBB resin to dentin after long-term water immersion.

    PubMed

    Soeno, Kohyoh; Taira, Yohsuke; Ito, Shuichi

    2012-01-01

    An experimental conditioner (Exp), which was an aqueous solution of 10% ascorbic acid and 5% ferric chloride, was prepared in this study. This study evaluated the effect of Exp on the microtensile bond strength between a self-curing resin and dentin after long-term water immersion. Flat human dentin surfaces were sequentially pretreated with 40% phosphoric acid, 10% sodium hypochlorite, and Exp. Surface pretreatment with an aqueous solution of 10% citric and 3% ferric chloride (10-3) was used as a control. Composite resin rods were bonded to pretreated dentin surfaces using 4-META/MMA-TBB resin. Microtensile bond strengths were evaluated after water immersion at 24 h, 12 months, 24 months, and 36 months. At each immersion period, the bond strength of Exp was significantly higher than that of 10-3. After 36 months, Exp showed no significant decrease in microtensile bond strength, but 10-3 showed significant reductions. Pretreatment with experimental acid/base conditioner markedly improved the bonding durability of 4-META/MMA-TBB resin to human dentin when compared against the conventional 10-3 treatment.

  10. The strength of an Ig switch region is determined by its ability to drive R loop formation and its number of WGCW sites.

    PubMed

    Zhang, Zheng Z; Pannunzio, Nicholas R; Han, Li; Hsieh, Chih-Lin; Yu, Kefei; Lieber, Michael R

    2014-07-24

    R loops exist at the murine IgH switch regions and possibly other locations, but their functional importance is unclear. In biochemical systems, R loop initiation requires DNA sequence regions containing clusters of G nucleotides, but cellular studies have not been done. Here, we vary the G-clustering, total switch region length, and the number of target sites (WGCW sites for the activation-induced deaminase) at synthetic switch regions in a murine B cell line to determine the effect on class switch recombination (CSR). G-clusters increase CSR regardless of their immediate proximity to the WGCW sites. This increase is accompanied by an increase in R loop formation. CSR efficiency correlates better with the absolute number of WGCW sites in the switch region rather than the total switch region length or density of WGCW sites. Thus, the overall strength of the switch region depends on G-clusters, which initiate R loop formation, and on the number of WGCW sites. PMID:25017067

  11. The Strength of an Ig Switch Region is Determined by its Ability to Drive R-loop Formation and its Number of WGCW Sites

    PubMed Central

    Zhang, Zheng Z.; Pannunzio, Nicholas R.; Han, Li; Hsieh, Chih-Lin; Yu, Kefei; Lieber, Michael R.

    2014-01-01

    SUMMARY R-loops exist at the murine IgH switch regions and possibly other locations, but their functional importance is unclear. In biochemical systems, R-loop initiation requires DNA sequence regions containing clusters of G nucleotides, but cellular studies have not been done. Here, we vary the G-clustering, total switch region length, and the number of target sites (WGCW sites for the activation-induced deaminase) at synthetic switch regions in a murine B cell line to determine the effect on class switch recombination (CSR). G-clusters increase CSR, regardless of their immediate proximity to the WGCW sites. This increase is accompanied by an increase in R-loop formation. CSR efficiency correlates better with the absolute number of WGCW sites in the switch region rather than the total switch region length or density of WGCW sites. Thus, the overall strength of the switch region depends on G-clusters, which initiate R-loop formation, and on the number of WGCW sites. PMID:25017067

  12. Recognition of protein phosphorylation site based on amino acids sequence features

    NASA Astrophysics Data System (ADS)

    Zhang, Ying; Ding, Changjiang; Lu, Jun

    2012-09-01

    Protein phosphorylation is one of the most important reversible post-translational modifications (PTMs), and the theoretical recognition of the phosphorylation site is one of the important content of the computational biology. In the paper, we use the amino acid component, the position-dependent residue statistics and the non-adjacent residue pair frequency as the recognition parameters, and use Jensen-Shannon Divergence with Quadratic Discriminant analysis(JSDQD) as the method for predicting the phosphorylation sites. The 7-fold cross-validation test accuracies for the CK2, PKA and PKC kinase families are 90%, 90% and 86%, respectively.

  13. Estimates of cloud water deposition at Mountain Acid Deposition Program sites in the Appalachian Mountains.

    PubMed

    Baumgardner, Ralph E; Isil, Selma S; Lavery, Thomas F; Rogers, Christopher M; Mohnen, Volker A

    2003-03-01

    Cloud water deposition was estimated at three high-elevation sites in the Appalachian Mountains of the eastern United States (Whiteface Mountain, NY; Whitetop Mountain, VA; and Clingman's Dome, TN) from 1994 through 1999 as part of the Mountain Acid Deposition Program (MADPro). This paper provides a summary of cloud water chemistry, cloud liquid water content, cloud frequency, estimates of cloud water deposition of sulfur and nitrogen species, and estimates of total deposition of sulfur and nitrogen at these sites. Other cloud studies in the Appalachians and their comparison to MADPro are also summarized. Whiteface Mountain exhibited the lowest mean and median concentrations of sulfur and nitrogen ions in cloud water, while Clingman's Dome exhibited the highest mean and median concentrations. This geographic gradient is partly an effect of the different meteorological conditions experienced at northern versus southern sites in addition to the difference in pollution content of air masses reaching the sites. All sites measured seasonal cloud water deposition rates of SO4(2-) greater than 50 kg/ha and NO3(-) rates of greater than 25 kg/ha. These high-elevation sites experienced additional deposition loading of SO4(2-) and NO3(-) on the order of 6-20 times greater compared with lower elevation Clean Air Status and Trends Network (CASTNet) sites. Approximately 80-90% of this extra loading is from cloud deposition.

  14. Reducing N2O Emission from a Domestic-Strength Nitrifying Culture by Free Nitrous Acid-Based Sludge Treatment.

    PubMed

    Wang, Dongbo; Wang, Qilin; Laloo, Andrew Elohim; Yuan, Zhiguo

    2016-07-19

    An increase of nitrite in the domestic-strength range is generally recognized to stimulate nitrous oxide (N2O) production by ammonia-oxidizing bacteria (AOB). It was found in this study, however, that N2O emission from a mainstream nitritation system (cyclic nitrite = 25-45 mg of N/L) that was established by free nitrous acid (FNA)-based sludge treatment was not higher but much lower than that from the initial nitrifying system with full conversion of NH4(+)-N to NO3(-)-N. Under dissolved oxygen (DO) levels of 2.5-3.0 mg/L, N2O emission from the nitritation stage was 76% lower than that from the initial stage. Even when the DO level was reduced to 0.3-0.8 mg/L, N2O emission from the nitritation stage was still 40% lower. An investigation of the mechanism showed that FNA treatment caused a shift of the stimulation threshold of nitrite on N2O emission. At the nitritation stage, the maximal N2O emission factor occurred at ∼16 mg of N/(L of nitrite). However, it increased with increasing nitrite in the range of 0-56 mg of N/L at the initial stage. FNA treatment decreased the biomass-specific N2O production rate, suggesting that the enzymes relevant to nitrifier denitrification were inhibited. Microbial analysis revealed that FNA treatment decreased the microbial community diversity but increased the abundances of AOB and denitrifiers. PMID:27294698

  15. Site-specific incorporation of keto amino acids into functional G protein-coupled receptors using unnatural amino acid mutagenesis.

    PubMed

    Ye, Shixin; Köhrer, Caroline; Huber, Thomas; Kazmi, Manija; Sachdev, Pallavi; Yan, Elsa C Y; Bhagat, Aditi; RajBhandary, Uttam L; Sakmar, Thomas P

    2008-01-18

    G protein-coupled receptors (GPCRs) are ubiquitous heptahelical transmembrane proteins involved in a wide variety of signaling pathways. The work described here on application of unnatural amino acid mutagenesis to two GPCRs, the chemokine receptor CCR5 (a major co-receptor for the human immunodeficiency virus) and rhodopsin (the visual photoreceptor), adds a new dimension to studies of GPCRs. We incorporated the unnatural amino acids p-acetyl-L-phenylalanine (Acp) and p-benzoyl-L-phenylalanine (Bzp) into CCR5 at high efficiency in mammalian cells to produce functional receptors harboring reactive keto groups at three specific positions. We obtained functional mutant CCR5, at levels up to approximately 50% of wild type as judged by immunoblotting, cell surface expression, and ligand-dependent calcium flux. Rhodopsin containing Acp at three different sites was also purified in high yield (0.5-2 microg/10(7) cells) and reacted with fluorescein hydrazide in vitro to produce fluorescently labeled rhodopsin. The incorporation of reactive keto groups such as Acp or Bzp into GPCRs allows their reaction with different reagents to introduce a variety of spectroscopic and other probes. Bzp also provides the possibility of photo-cross-linking to identify precise sites of protein-protein interactions, including GPCR binding to G proteins and arrestins, and for understanding the molecular basis of ligand recognition by chemokine receptors. PMID:17993461

  16. Ab initio periodic Hartree-Fock investigation of a zeolite acid site

    SciTech Connect

    Nicholas, J.B.; Hess, A.C. )

    1994-06-15

    We have studied theoretical models of a zeolite Bronsted acid site using ab initio periodic Hartree-Fock (PHF) theory. We present results for both one and two acid sites inside a sodalite structure ([beta]-cage). We optimized the geometry of the bridging hydroxyl groups using the STO-3G basis set. All other reported crystal properties were evaluated using the more complete 6-21G (in which the outermost orbital exponents were reoptimized for the solid) level of theory. The results of the periodic calculations are compared to available experimental data, similar periodic calculations of silica sodalite in which there are no acid sites, and calculations of clusters that mimic parts of the periodic structure. The optimized geometry of the single bridging hydroxyl group has internal coordinates of Si-O = 1.64 [angstrom], Al-O = 1.72 [angstrom], O-H = 0.97 [angstrom], Si-O(H)-Al = 136.3[degree], and Si-O-H = 110.7[degree], and the hydrogen is bent 25.2[degrees] out of the Si-O-Al plane. The optimized geometry of the bridging hydroxyl group in the two-acid site model is very similar, with Si-O = 1.65 [angstrom], Al-O = 1.72 [angstrom], O-H = 0.97 [angstrom], Si-O(H)-Al = 135.5[degree], Si-O-H = 112.7[degree], and an out-of-plane angle of 21.5[degree]. 42 refs., 11 figs., 5 tabs.

  17. Disulfide bridge regulates ligand-binding site selectivity in liver bile acid-binding proteins.

    PubMed

    Cogliati, Clelia; Tomaselli, Simona; Assfalg, Michael; Pedò, Massimo; Ferranti, Pasquale; Zetta, Lucia; Molinari, Henriette; Ragona, Laura

    2009-10-01

    Bile acid-binding proteins (BABPs) are cytosolic lipid chaperones that play central roles in driving bile flow, as well as in the adaptation to various pathological conditions, contributing to the maintenance of bile acid homeostasis and functional distribution within the cell. Understanding the mode of binding of bile acids with their cytoplasmic transporters is a key issue in providing a model for the mechanism of their transfer from the cytoplasm to the nucleus, for delivery to nuclear receptors. A number of factors have been shown to modulate bile salt selectivity, stoichiometry, and affinity of binding to BABPs, e.g. chemistry of the ligand, protein plasticity and, possibly, the formation of disulfide bridges. Here, the effects of the presence of a naturally occurring disulfide bridge on liver BABP ligand-binding properties and backbone dynamics have been investigated by NMR. Interestingly, the disulfide bridge does not modify the protein-binding stoichiometry, but has a key role in modulating recognition at both sites, inducing site selectivity for glycocholic and glycochenodeoxycholic acid. Protein conformational changes following the introduction of a disulfide bridge are small and located around the inner binding site, whereas significant changes in backbone motions are observed for several residues distributed over the entire protein, both in the apo form and in the holo form. Site selectivity appears, therefore, to be dependent on protein mobility rather than being governed by steric factors. The detected properties further establish a parallelism with the behaviour of human ileal BABP, substantiating the proposal that BABPs have parallel functions in hepatocytes and enterocytes. PMID:19754879

  18. Disulfide bridge regulates ligand-binding site selectivity in liver bile acid-binding proteins.

    PubMed

    Cogliati, Clelia; Tomaselli, Simona; Assfalg, Michael; Pedò, Massimo; Ferranti, Pasquale; Zetta, Lucia; Molinari, Henriette; Ragona, Laura

    2009-10-01

    Bile acid-binding proteins (BABPs) are cytosolic lipid chaperones that play central roles in driving bile flow, as well as in the adaptation to various pathological conditions, contributing to the maintenance of bile acid homeostasis and functional distribution within the cell. Understanding the mode of binding of bile acids with their cytoplasmic transporters is a key issue in providing a model for the mechanism of their transfer from the cytoplasm to the nucleus, for delivery to nuclear receptors. A number of factors have been shown to modulate bile salt selectivity, stoichiometry, and affinity of binding to BABPs, e.g. chemistry of the ligand, protein plasticity and, possibly, the formation of disulfide bridges. Here, the effects of the presence of a naturally occurring disulfide bridge on liver BABP ligand-binding properties and backbone dynamics have been investigated by NMR. Interestingly, the disulfide bridge does not modify the protein-binding stoichiometry, but has a key role in modulating recognition at both sites, inducing site selectivity for glycocholic and glycochenodeoxycholic acid. Protein conformational changes following the introduction of a disulfide bridge are small and located around the inner binding site, whereas significant changes in backbone motions are observed for several residues distributed over the entire protein, both in the apo form and in the holo form. Site selectivity appears, therefore, to be dependent on protein mobility rather than being governed by steric factors. The detected properties further establish a parallelism with the behaviour of human ileal BABP, substantiating the proposal that BABPs have parallel functions in hepatocytes and enterocytes.

  19. Effect of hybridization on bond strength and adhesive interface after acid-base challenge using 4-META/MMA-TBB resin.

    PubMed

    Takagaki, Tomohiro; Nikaido, Toru; Tsuchiya, Satoko; Ikeda, Masaomi; Foxton, Richard M; Tagami, Junji

    2009-03-01

    The purposes of this study were twofold, namely to evaluate: (1) the effect of hybridization on microtensile bond strength (microTBS) to dentin, and (2) the ultrastructure of the dentin-adhesive interface with 4-META/MMA-TBB resin after acid-base challenge. Dentin surfaces, which received no treatment (NT), 65% phosphoric acid (PA), or 10% citric acid-3% ferric chloride (10-3), were bonded with a 4-META/MMA-TBB resin. To evaluate dentin bond strength, microTBS test was performed at a crosshead speed of 1 mm/min. For ultrastructural evaluation of the adhesive interfaces, SEM was used to examine the interfaces of the bonded specimens after acid-base challenge. The microTBS of NT was not determined, while that of 10-3 was significantly higher than that of PA (p < 0.05). With PA and 10-3, the hybrid layer was clearly observed, but no so for the acid-base resistant zone. Wall lesion was found in NT only. In conclusion, hybridization is vital to improving microTBS to dentin and enhancing resistance at the adhesive interface against acid-base challenge.

  20. Micro-shear bond strength and surface micromorphology of a feldspathic ceramic treated with different cleaning methods after hydrofluoric acid etching

    PubMed Central

    STEINHAUSER, Henrique Caballero; TURSSI, Cecília Pedroso; FRANÇA, Fabiana Mantovani Gomes; do AMARAL, Flávia Lucisano Botelho; BASTING, Roberta Tarkany

    2014-01-01

    Objective The aim of this study was to evaluate the effect of feldspathic ceramic surface cleaning on micro-shear bond strength and ceramic surface morphology. Material and Methods Forty discs of feldspathic ceramic were prepared and etched with 10% hydrofluoric acid for 2 minutes. The discs were randomly distributed into five groups (n=8): C: no treatment, S: water spray + air drying for 1 minute, US: immersion in ultrasonic bath for 5 minutes, F: etching with 37% phosphoric acid for 1 minute, followed by 1-minute rinse, F+US: etching with 37% phosphoric acid for 1 minute, 1-minute rinse and ultrasonic bath for 5 minutes. Composite cylinders were bonded to the discs following application of silane and hydrophobic adhesive for micro-shear bond strength testing in a universal testing machine at 0.5 mm/min crosshead speed until failure. Stereomicroscopy was used to classify failure type. Surface micromorphology of each treatment type was evaluated by scanning electron microscopy at 500 and 2,500 times magnification. Results One-way ANOVA test showed no significant difference between treatments (p=0.3197) and the most common failure types were cohesive resin cohesion followed by adhesive failure. Micro-shear bond strength of the feldspathic ceramic substrate to the adhesive system was not influenced by the different surface cleaning techniques. Absence of or less residue was observed after etching with hydrofluoric acid for the groups US and F+US. Conclusions Combining ceramic cleaning techniques with hydrofluoric acid etching did not affect ceramic bond strength, whereas, when cleaning was associated with ultrasound, less residue was observed. PMID:24676577

  1. Remaining Sites Verification Package for the 120-B-1, 105-B Battery Acid Sump, Waste Site Reclassification Form 2006-057

    SciTech Connect

    L. M. Dittmer

    2006-09-25

    The 120-B-1 waste site, located in the 100-BC-1 Operable Unit of the Hanford Site, consisted of a concrete battery acid sump that was used from 1944 to 1969 to neutralize the spent sulfuric acid from lead cell batteries of emergency power packs and the emergency lighting system. The battery acid sump was associated with the 105-B Reactor Building and was located adjacent to the building's northwest corner. The results of verification sampling demonstrated that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results also showed that residual contaminant concentrations are protective of groundwater and the Columbia River.

  2. Surface site density, silicic acid retention and transport properties of compacted magnetite powder

    NASA Astrophysics Data System (ADS)

    Mayant, C.; Grambow, B.; Abdelouas, A.; Ribet, S.; Leclercq, S.

    In France, within the framework of investigations of the feasibility of deep geological disposal of high-level radioactive waste, studies on corrosion products of steel over packs are ongoing. Such studies concern silica and radionuclide retention. The objective of the present work is to study sorption of silicic acid on compacted magnetite in percolation cells to attempt to simulate confined site conditions. Potentiometric titration of commercial magnetite was carried out with both dispersed and compacted magnetite. The titration of the magnetite suspension has been made with two different methods: a batch method (several suspensions) and a direct fast method (one suspension). The Gran’s function gave 1.7 (±0.4) and 2.4 (±0.5) sorption sites nm -2 with these respective methods but site densities as high as 20/nm 2 could be obtained by modelling. The titration of magnetite compacted at 120 bars showed that the evolution of charge density on magnetite surfaces is similar for compacted and dispersed magnetite. Silicic acid sorption onto dispersed and compacted magnetite was similar with sorption site densities ranging between 2.2 and 4.4/nm 2.

  3. Ionization-site effects on the photofragmentation of chloro- and bromoacetic acid molecules

    NASA Astrophysics Data System (ADS)

    Levola, Helena; Itälä, Eero; Schlesier, Kim; Kooser, Kuno; Laine, Sanna; Laksman, Joakim; Ha, Dang Trinh; Rachlew, Elisabeth; Tarkanovskaja, Marta; Tanzer, Katrin; Kukk, Edwin

    2015-12-01

    Fragmentation of gas-phase chloro- and bromoacetic acid samples, particularly its dependency on the atomic site of the initial core ionization, was studied in photoelectron-photoion-photoion coincidence (PEPIPICO) measurements. The fragmentation was investigated after ionizing carbon 1 s and bromine 3 d or chlorine 2 p core orbitals. It was observed that the samples had many similar fragmentation pathways and that their relative weights depended strongly on the initial ionization site. Additional Auger PEPIPICO measurements revealed a clear dependence of fragment pair intensities on the kinetic energy of the emitted Auger electrons. The modeled and measured Auger electron spectra indicated that the average internal energy of the molecule was larger following the carbon 1 s core-hole decay than after the decay of the halogen core hole. This difference in the internal energies was found to be the source of the site-dependent photofragmentation behavior.

  4. Influence of high-fat diet from differential dietary sources on bone mineral density, bone strength, and bone fatty acid composition in rats.

    PubMed

    Lau, Beatrice Y; Fajardo, Val Andrew; McMeekin, Lauren; Sacco, Sandra M; Ward, Wendy E; Roy, Brian D; Peters, Sandra J; Leblanc, Paul J

    2010-10-01

    Previous studies have suggested that high-fat diets adversely affect bone development. However, these studies included other dietary manipulations, including low calcium, folic acid, and fibre, and (or) high sucrose or cholesterol, and did not directly compare several common sources of dietary fat. Thus, the overall objective of this study was to investigate the effect of high-fat diets that differ in fat quality, representing diets high in saturated fatty acids (SFA), n-3 polyunsaturated fatty acids (PUFA), or n-6 PUFA, on femur bone mineral density (BMD), strength, and fatty acid composition. Forty-day-old male Sprague-Dawley rats were maintained for 65 days on high-fat diets (20% by weight), containing coconut oil (SFA; n = 10), flaxseed oil (n-3 PUFA; n = 10), or safflower oil (n-6 PUFA; n = 11). Chow-fed rats (n = 10), at 105 days of age, were included to represent animals on a control diet. Rats fed high-fat diets had higher body weights than the chow-fed rats (p < 0.001). Among all high-fat groups, there were no differences in femur BMD (p > 0.05) or biomechanical strength properties (p > 0.05). Femurs of groups fed either the high n-3 or high n-6 PUFA diets were stronger (as measured by peak load) than those of the chow-fed group, after adjustment for significant differences in body weight (p = 0.001). As expected, the femur fatty acid profile reflected the fatty acid composition of the diet consumed. These results suggest that high-fat diets, containing high levels of PUFA in the form of flaxseed or safflower oil, have a positive effect on bone strength when fed to male rats 6 to 15 weeks of age.

  5. An improved thermodynamic model for the complexation of trivalent actinides and lanthanide with oxalic acid valid to high ionic strength.

    SciTech Connect

    Xiong, Yongliang; Thakur, Punam; Borkowski, Marian

    2015-07-30

    The dissociation constants of oxalic acid (Ox), and the stability constants of Am3+, Cm3+ and Eu3+ with Ox2– have been determined at 25 °C, over a range of concentration varying from 0.1 to 6.60 m NaClO4 using potentiometric titration and extraction techniques, respectively. The experimental data support the formation of complexes, M(Ox)n3 – 2n, where (M = Am3+, Cm3+ and Eu3+ and n = 1 and 2). The dissociation constant and the stability constant values measured as a function of NaClO4 concentration were used to estimate the Pitzer parameters for the respective interactions of Am3+, Cm3+ and Eu3+ with Ox. Furthermore, the stability constants data of Am3+ –Ox measured in NaClO4 and in NaCl solutions from the literature were simultaneously fitted in order to refine the existing actinide–oxalate complexation model that can be used universally in the safety assessment of radioactive waste disposal. The thermodynamic stability constant: log β0101 = 6.30 ± 0.06 and log β0102 = 10.84 ± 0.06 for Am3+ was obtained by simultaneously fitting data in NaCl and NaClO4 media. Additionally, log β0101 = 6.72 ± 0.08 and log β0102 = 11.05 ± 0.09 for the Cm3+ and log β0101 = 6.67 ± 0.08 and log β0102 = 11.15 ± 0.09 for the Eu3+ were calculated by extrapolation of data to zero ionic strength in NaClO4 medium only. For all stability constants, the Pitzer model gives an excellent representation of the data using interaction parameters β(0), β(1), and CΦ determined in this work. The thermodynamic model developed in this work will be useful in accurately modeling the potential solubility of trivalent

  6. Microbial Communities in Biofilms of an Acid Mine Drainage Site Determined by Phospholipid Analysis

    NASA Astrophysics Data System (ADS)

    Das Gupta, S.; Fang, J.

    2008-12-01

    Phospholipids were extracted to determine the microbial biomass and community structure of biofims from an acid mine drainage (AMD) at the Green Valley coal mine site (GVS) in western Indiana. The distribution of specific biomarkers indicated the presence of a variety of microorganisms. Phototrophic microeukaryotes, which include Euglena mutabilis, algae, and cyanobacteria were the most dominant organisms, as indicated by the presence of polyunsaturated fatty acids. The presence of terminally methyl branched fatty acids suggests the presence of Gram-positive bacteria, and the mid-methyl branched fatty acids indicates the presence of sulfate-reducing bacteria. Fungi appear to also be an important part of the AMD microbial communities as suggested by the presence of 18:2 fatty acid. The acidophilic microeukaryotes Euglena dominated the biofilm microbial communities. These microorganisms appear to play a prominent role in the formation and preservation of stromatolites and in releasing oxygen to the atmosphere by oxygenic photosynthesis. Thus, the AMD environment comprises a host of microorganisms spreading out within the phylogenetic tree of life. Novel insights on the roles of microbial consortia in the formation and preservation of stromatolites and the production of oxygen through photosynthesis in AMD systems may have significance in the understanding of the interaction of Precambrian microbial communities in environments that produced microbially-mediated sedimentary structures and that caused oxygenation of Earth's atmosphere.

  7. Oxygen-enhanced biodegradation of phenoxy acids in ground water at contaminated sites.

    PubMed

    Tuxen, Nina; Reitzel, Lotte A; Albrechtsen, Hans-Jørgen; Bjerg, Poul L

    2006-01-01

    The effects of adding oxygen to anaerobic aquifer materials on biodegradation of phenoxy acid herbicides were studied by laboratory experiments with aquifer material from two contaminated sites (a former agricultural machinery service and an old landfill). At both sites, the primary pollutants were phenoxy acids and related chlorophenols. It was found that addition of oxygen enhanced degradation of the six original phenoxy acids and six original chlorophenols. Inverse modeling on 14C 4-chloro-2-methylphenoxypropanoic acid (MCPP) degradation curves revealed that increasing the oxygen concentrations from <0.3 mg/L up to 7 to 8 mg/L shortened the lag phases (from approximately 150 d to 5 to 25 d) and increased first-order degradation rate constants by 1 order of magnitude (from approximately 5 x 10(-2) d(-1) to up to 30 x 10(-2) d(-1)). Additionally, the degree of MCPP mineralization was increased (30% to 50% mineralized at low oxygen concentrations and 50% to 70% mineralized at high oxygen concentrations, based on 14CO2 recovery). These positive effects on degradation were observed even at relatively low oxygen concentrations (2 mg/L). Furthermore, effects related to the addition of oxygen on the general geochemistry were studied. An oxygen consumption of 2.2 to 2.6 mg O2/g dw was observed due to oxidation of solid organic matter and, to some extent (0.5% to 11% of the total oxygen consumption), water-soluble compounds such as Fe2+, dissolved Mn, nonvolatile organic carbon, and NH4+. Overall, the results suggest that stimulated biodegradation by addition of oxygen might be a feasible remediation technology at herbicide-contaminated sites, although oxygen consumption by the sediment could limit the applicability.

  8. Characterizing Surface Acidic Sites in Mesoporous-Silica-Supported Tungsten Oxide Catalysts Using Solid State NMR and Quantum Chemistry Calculations

    SciTech Connect

    Hu, Jian Z.; Kwak, Ja Hun; Wang, Yong; Hu, Mary Y.; Turcu, Romulus VF; Peden, Charles HF

    2011-10-18

    The acidic sites in dispersed tungsten oxide supported on SBA-15 mesoporous silica were investigated using a combination of pyridine titration, both fast-, and slow-MAS {sup 15}N NMR, static {sup 2}H NMR, and quantum chemistry calculations. It is found that the bridged acidic -OH groups in surface adsorbed tungsten dimers (i.e., W-OH-W) are the Broensted acid sites. The unusually strong acidity of these Broensted acid sites is confirmed by quantum chemistry calculations. In contrast, terminal W-OH sites are very stable and only weakly acidic as are terminal Si-OH sites. Furthermore, molecular interactions between pyridine molecules and the dimer Broensted and terminal W-OH sites for dispersed tungsten oxide species is strong. This results in restricted molecular motion for the interacting pyridine molecules even at room temperature, i.e., a reorientation mainly about the molecular 2-fold axis. This restricted reorientation makes it possible to estimate the relative ratio of the Broensted (tungsten dimer) to the weakly acidic terminal W-OH sites in the catalyst using the slow-MAS {sup 1}H-{sup 15}N CP PASS method.

  9. Comparison of shear bond strength of composite resin to enamel surface with laser etching versus acid etching: An in vitro evaluation

    PubMed Central

    Hoshing, Upendra A; Patil, Suvarna; Medha, Ashish; Bandekar, Siddhesh Dattatray

    2014-01-01

    Introduction: The aim of the study is in vitro evaluation of the shear bond strength of composite resin bonded to enamel which is pretreated using acid etchant and Er,Cr:Ysgg. Materials and Methods: 40 extracted human teeth were divided in two groups of 20 each (Groups A and B). In Group A, prepared surface of enamel was etched using 37% phosphoric acid (Scotchbond, 3M). In Group B, enamel was surface treated by a an Er, Cr: YSGG laser system (Waterlase MD, Biolase Technology Inc., San Clemente, CA, USA) operating at a wavelength of 2,780 nm and having a pulse duration of 140-200 microsecond with a repetition rate of 20 Hz and 40 Hz. Bonding agent ((Scotchbond Multipurpose, 3M) was applied over the test areas on 20 samples of Groups A and B each, and light cured. Composite resin (Ceram X duo Nanoceramic restorative, Densply) was applied onto the test areas as a 3 × 3 mm diameter bid, and light cured. The samples were tested for shear bond strength. Results: Mean shear bond strength for acid-etched enamel (26.41 ± 0.66MPa, range 25.155 to 27.150 MPa) was significantly higher (P < 0.01) than for laser-etched enamel (16.23 ± 0.71MPa, range 15.233 to 17.334 MPa). Conclusions: For enamel surface, mean shear bond strength of bonded composite obtained after laser etching were significantly lower than those obtained after acid etching. PMID:25125842

  10. The Effect of an Acidic Food-Simulating Environment on the Shear Bond Strength of Self-Ligating Brackets with Different Base Designs

    PubMed Central

    Sheibaninia, Ahmad; Sepasi, Sepehr; Saghiri, Mohammad Ali; Sepasi, Setareh

    2014-01-01

    Aim. This study aims to evaluate the effect of acidic food simulant and (acetic acid 3%) on the shear bond strength (SBS) and adhesive remnant index (ARI) scores of one conventional and three different self-ligating brackets with different base designs. Materials and Methods. Freshly extracted first maxillary premolars (n = 160) were embedded in resin blocks. A conventional stainless steel bracket, Equilibrium 2, and three types of self-ligating brackets, Speed, In-Ovation R, and Damon 3MX, were bonded to teeth and exposed to distilled water (groups 1, 3, 5, and 7) or acetic acid 3% (groups 2, 4, 6, 8) for 12 weeks. SBS and ARI were calculated and statistical analysis was performed with the analysis of variance (SBS) or χ2 test (ARI) to compare values between the different groups. Results. Equilibrium 2 and In-Ovation R showed a significantly lower SBS in the acidic environment than in distilled water. Significant differences in ARI scores were found for Equilibrium 2 after immersion in an acidic environment, shifting from 0 in distilled water to 2 in an acidic environment. Conclusions. Equilibrium 2 and In-Ovation R brackets showed a significant decrease in SBS after a 12-week immersion in acetic acid 3%, although all groups showed clinically acceptable SBS. Equilibrium 2 showed significant differences in ARI scores when exposed to acetic acid 3%. PMID:25328524

  11. Crystal Structure of Species D Adenovirus Fiber Knobs and Their Sialic Acid Binding Sites

    PubMed Central

    Burmeister, Wim P.; Guilligay, Delphine; Cusack, Stephen; Wadell, Göran; Arnberg, Niklas

    2004-01-01

    Adenovirus serotype 37 (Ad37) belongs to species D and can cause epidemic keratoconjunctivitis, whereas the closely related Ad19p does not. Primary cell attachment by adenoviruses is mediated through receptor binding of the knob domain of the fiber protein. The knobs of Ad37 and Ad19p differ at only two positions, Lys240Glu and Asn340Asp. We report the high-resolution crystal structures of the Ad37 and Ad19p knobs, both native and in complex with sialic acid, which has been proposed as a receptor for Ad37. Overall, the Ad37 and Ad19p knobs are very similar to previously reported knob structures, especially to that of Ad5, which binds the coxsackievirus-adenovirus receptor (CAR). Ad37 and Ad19p knobs are structurally identical with the exception of the changed side chains and are structurally most similar to CAR-binding knobs (e.g., that of Ad5) rather than non-CAR-binding knobs (e.g., that of Ad3). The two mutations in Ad19p result in a partial loss of the exceptionally high positive surface charge of the Ad37 knob but do not affect sialic acid binding. This site is located on the top of the trimer and binds both α(2,3) and α(2,6)-linked sialyl-lactose, although only the sialic acid residue makes direct contact. Amino acid alignment suggests that the sialic acid binding site is conserved in several species D serotypes. Our results show that the altered viral tropism and cell binding of Ad19p relative to those of Ad37 are not explained by a different binding ability toward sialyl-lactose. PMID:15220447

  12. Computational study on the roles of amino acid residues in the active site formation mechanism of blue-light photoreceptors

    NASA Astrophysics Data System (ADS)

    Sato, Ryuma; Kitoh-Nishioka, Hirotaka; Ando, Koji; Yamato, Takahisa

    2015-07-01

    To examine the functional roles of the active site methionine (M-site) and glutamic acid (E-site) residues of blue-light photoreceptors, we performed in silico mutation at the M-site in a systematic manner and focused on the hydrogen bonding between the E-site and the substrate: the cyclobutane-pyrimidine dimer (CPD). Fragment molecular orbital calculations with electron correlations demonstrated that substitution of the M-site methionine with either alanine or glutamine always destabilizes the interaction energy between the E-site and the CPD by more than 12.0 kcal/mol, indicating that the methionine and glutamic acid residues cooperatively facilitate the enzymatic reaction in the active site.

  13. A Large-Scale Assessment of Nucleic Acids Binding Site Prediction Programs

    PubMed Central

    Miao, Zhichao; Westhof, Eric

    2015-01-01

    Computational prediction of nucleic acid binding sites in proteins are necessary to disentangle functional mechanisms in most biological processes and to explore the binding mechanisms. Several strategies have been proposed, but the state-of-the-art approaches display a great diversity in i) the definition of nucleic acid binding sites; ii) the training and test datasets; iii) the algorithmic methods for the prediction strategies; iv) the performance measures and v) the distribution and availability of the prediction programs. Here we report a large-scale assessment of 19 web servers and 3 stand-alone programs on 41 datasets including more than 5000 proteins derived from 3D structures of protein-nucleic acid complexes. Well-defined binary assessment criteria (specificity, sensitivity, precision, accuracy…) are applied. We found that i) the tools have been greatly improved over the years; ii) some of the approaches suffer from theoretical defects and there is still room for sorting out the essential mechanisms of binding; iii) RNA binding and DNA binding appear to follow similar driving forces and iv) dataset bias may exist in some methods. PMID:26681179

  14. A Large-Scale Assessment of Nucleic Acids Binding Site Prediction Programs.

    PubMed

    Miao, Zhichao; Westhof, Eric

    2015-12-01

    Computational prediction of nucleic acid binding sites in proteins are necessary to disentangle functional mechanisms in most biological processes and to explore the binding mechanisms. Several strategies have been proposed, but the state-of-the-art approaches display a great diversity in i) the definition of nucleic acid binding sites; ii) the training and test datasets; iii) the algorithmic methods for the prediction strategies; iv) the performance measures and v) the distribution and availability of the prediction programs. Here we report a large-scale assessment of 19 web servers and 3 stand-alone programs on 41 datasets including more than 5000 proteins derived from 3D structures of protein-nucleic acid complexes. Well-defined binary assessment criteria (specificity, sensitivity, precision, accuracy…) are applied. We found that i) the tools have been greatly improved over the years; ii) some of the approaches suffer from theoretical defects and there is still room for sorting out the essential mechanisms of binding; iii) RNA binding and DNA binding appear to follow similar driving forces and iv) dataset bias may exist in some methods. PMID:26681179

  15. RF1 knockout allows ribosomal incorporation of unnatural amino acids at multiple sites.

    PubMed

    Johnson, David B F; Xu, Jianfeng; Shen, Zhouxin; Takimoto, Jeffrey K; Schultz, Matthew D; Schmitz, Robert J; Xiang, Zheng; Ecker, Joseph R; Briggs, Steven P; Wang, Lei

    2011-11-01

    Stop codons have been exploited for genetic incorporation of unnatural amino acids (Uaas) in live cells, but their low incorporation efficiency, which is possibly due to competition from release factors, limits the power and scope of this technology. Here we show that the reportedly essential release factor 1 (RF1) can be knocked out from Escherichia coli by 'fixing' release factor 2 (RF2). The resultant strain JX33 is stable and independent, and it allows UAG to be reassigned from a stop signal to an amino acid when a UAG-decoding tRNA-synthetase pair is introduced. Uaas were efficiently incorporated at multiple UAG sites in the same gene without translational termination in JX33. We also found that amino acid incorporation at endogenous UAG codons is dependent on RF1 and mRNA context, which explains why E. coli tolerates apparent global suppression of UAG. JX33 affords a unique autonomous host for synthesizing and evolving new protein functions by enabling Uaa incorporation at multiple sites. PMID:21926996

  16. Identification of a lithium interaction site in the gamma-aminobutyric acid (GABA) transporter GAT-1.

    PubMed

    Zhou, Yonggang; Zomot, Elia; Kanner, Baruch I

    2006-08-01

    The sodium- and chloride-dependent electrogenic gamma-aminobutyric acid (GABA) transporter GAT-1, which transports two sodium ions together with GABA, is essential for synaptic transmission by this neurotransmitter. Although lithium by itself does not support GABA transport, it has been proposed that lithium can replace sodium at one of the binding sites but not at the other. To identify putative lithium selectivity determinants, we have mutated the five GAT-1 residues corresponding to those whose side chains participate in the sodium binding sites Na1 and Na2 of the bacterial leucine-transporting homologue LeuT(Aa). In GAT-1 and in most other neurotransmitter transporter family members, four of these residues are conserved, but aspartate 395 replaces the Na2 residue threonine 354. At varying extracellular sodium, lithium stimulated sodium-dependent transport currents as well as [3H]GABA uptake in wild type GAT-1. The extent of this stimulation was dependent on the GABA concentration. In mutants in which aspartate 395 was replaced by threonine or serine, the stimulation of transport by lithium was abolished. Moreover, these mutants were unable to mediate the lithium leak currents. This phenotype was not observed in mutants at the four other positions, although their transport properties were severely impacted. Thus at saturating GABA, the site corresponding to Na2 behaves as a low affinity sodium binding site where lithium can replace sodium. We propose that GABA participates in the other sodium binding site, just like leucine does in the Na1 site, and that at limiting GABA, this site determines the apparent sodium affinity of GABA transport.

  17. Modelling metal centres, acid sites and reaction mechanisms in microporous catalysts.

    PubMed

    O'Malley, Alexander J; Logsdail, A J; Sokol, A A; Catlow, C R A

    2016-07-01

    We discuss the role of QM/MM (embedded cluster) computational techniques in catalytic science, in particular their application to microporous catalysis. We describe the methodologies employed and illustrate their utility by briefly summarising work on metal centres in zeolites. We then report a detailed investigation into the behaviour of methanol at acidic sites in zeolites H-ZSM-5 and H-Y in the context of the methanol-to-hydrocarbons/olefins process. Studying key initial steps of the reaction (the adsorption and subsequent methoxylation), we probe the effect of framework topology and Brønsted acid site location on the energetics of these initial processes. We find that although methoxylation is endothermic with respect to the adsorbed system (by 17-56 kJ mol(-1) depending on the location), there are intriguing correlations between the adsorption/reaction energies and the geometries of the adsorbed species, of particular significance being the coordination of methyl hydrogens. These observations emphasise the importance of adsorbate coordination with the framework in zeolite catalysed conversions, and how this may vary with framework topology and site location, particularly suited to investigation by QM/MM techniques. PMID:27136967

  18. Interaction of Beta-Hydroxy-Beta-Methylbutyrate Free Acid and Adenosine Triphosphate on Muscle Mass, Strength, and Power in Resistance Trained Individuals.

    PubMed

    Lowery, Ryan P; Joy, Jordan M; Rathmacher, John A; Baier, Shawn M; Fuller, John C; Shelley, Mack C; Jäger, Ralf; Purpura, Martin; Wilson, Stephanie M C; Wilson, Jacob M

    2016-07-01

    Lowery, RP, Joy, JM, Rathmacher, JA, Baier, SM, Fuller, JC Jr, Shelley, MC II, Jäger, R, Purpura, M, Wilson, SMC, and Wilson, JM. Interaction of beta-hydroxy-beta-methylbutyrate free acid and adenosine triphosphate on muscle mass, strength, and power in resistance trained individuals. J Strength Cond Res 30(7): 1843-1854, 2016-Adenosine-5'-triphosphate (ATP) supplementation helps maintain performance under high fatiguing contractions and with greater fatigue recovery demands also increase. Current evidence suggests that the free acid form of β-hydroxy-β-methylbutyrate (HMB-FA) acts by speeding regenerative capacity of skeletal muscle after high-intensity or prolonged exercise. Therefore, we investigated the effects of 12 weeks of HMB-FA (3 g) and ATP (400 mg) administration on lean body mass (LBM), strength, and power in trained individuals. A 3-phase double-blind, placebo-, and diet-controlled study was conducted. Phases consisted of an 8-week periodized resistance training program (phase 1), followed by a 2-week overreaching cycle (phase 2), and a 2-week taper (phase 3). Lean body mass was increased by a combination of HMB-FA/ATP by 12.7% (p < 0.001). In a similar fashion, strength gains after training were increased in HMB-FA/ATP-supplemented subjects by 23.5% (p < 0.001). Vertical jump and Wingate power were increased in the HMB-FA/ATP-supplemented group compared with the placebo-supplemented group, and the 12-week increases were 21.5 and 23.7%, respectively. During the overreaching cycle, strength and power declined in the placebo group (4.3-5.7%), whereas supplementation with HMB-FA/ATP resulted in continued strength gains (1.3%). In conclusion, HMB-FA and ATP in combination with resistance exercise training enhanced LBM, power, and strength. In addition, HMB-FA plus ATP blunted the typical response to overreaching, resulting in a further increase in strength during that period. It seems that the combination of HMB-FA/ATP could benefit those who

  19. Mutational Studies on Resurrected Ancestral Proteins Reveal Conservation of Site-Specific Amino Acid Preferences throughout Evolutionary History

    PubMed Central

    Risso, Valeria A.; Manssour-Triedo, Fadia; Delgado-Delgado, Asunción; Arco, Rocio; Barroso-delJesus, Alicia; Ingles-Prieto, Alvaro; Godoy-Ruiz, Raquel; Gavira, Jose A.; Gaucher, Eric A.; Ibarra-Molero, Beatriz; Sanchez-Ruiz, Jose M.

    2015-01-01

    Local protein interactions (“molecular context” effects) dictate amino acid replacements and can be described in terms of site-specific, energetic preferences for any different amino acid. It has been recently debated whether these preferences remain approximately constant during evolution or whether, due to coevolution of sites, they change strongly. Such research highlights an unresolved and fundamental issue with far-reaching implications for phylogenetic analysis and molecular evolution modeling. Here, we take advantage of the recent availability of phenotypically supported laboratory resurrections of Precambrian thioredoxins and β-lactamases to experimentally address the change of site-specific amino acid preferences over long geological timescales. Extensive mutational analyses support the notion that evolutionary adjustment to a new amino acid may occur, but to a large extent this is insufficient to erase the primitive preference for amino acid replacements. Generally, site-specific amino acid preferences appear to remain conserved throughout evolutionary history despite local sequence divergence. We show such preference conservation to be readily understandable in molecular terms and we provide crystallographic evidence for an intriguing structural-switch mechanism: Energetic preference for an ancestral amino acid in a modern protein can be linked to reorganization upon mutation to the ancestral local structure around the mutated site. Finally, we point out that site-specific preference conservation naturally leads to one plausible evolutionary explanation for the existence of intragenic global suppressor mutations. PMID:25392342

  20. The synergic effect between Mo species and acid sites in Mo/HMCM-22 catalysts for methane aromatization.

    PubMed

    Ma, Ding; Zhu, Qingjun; Wu, Zili; Zhou, Danhong; Shu, Yuying; Xin, Qin; Xu, Yide; Bao, Xinhe

    2005-08-21

    The acid properties of Mo/HMCM-22 catalyst, which is the precursor form of the working catalyst for methane aromatization reaction, and the synergic effect between Mo species and acid sites were studied and characterized by various characterization techniques. It is concluded that Brønsted and Lewis acidities of HMCM-22 are modified due to the introduction of molybdenum. We suggest a monomer of Mo species is formed by the exchange of Mo species with the Brønsted acid sites. On the other hand, coordinate unsaturated sites (CUS) are suggested to be responsible for the formation of newly detected Lewis acid sites. Computer modelling is established and coupling with experimental results, it is then speculated that the effective activation of methane is properly accomplished on Mo species accommodated in the 12 MR supercages of MCM-22 zeolite whereas the Brønsted acid sites in the same channel system play a key role for the formation of benzene. A much more pronounced volcano-typed reactivity curve of the Mo/HMCM-22 catalysts, as compared with that of the Mo/HZSM-5, with respect to Mo loading is found and this can be well understood due to the unique channel structure of MCM-22 zeolite and synergic effect between Mo species and acid sites.

  1. Mutational studies on resurrected ancestral proteins reveal conservation of site-specific amino acid preferences throughout evolutionary history.

    PubMed

    Risso, Valeria A; Manssour-Triedo, Fadia; Delgado-Delgado, Asunción; Arco, Rocio; Barroso-delJesus, Alicia; Ingles-Prieto, Alvaro; Godoy-Ruiz, Raquel; Gavira, Jose A; Gaucher, Eric A; Ibarra-Molero, Beatriz; Sanchez-Ruiz, Jose M

    2015-02-01

    Local protein interactions ("molecular context" effects) dictate amino acid replacements and can be described in terms of site-specific, energetic preferences for any different amino acid. It has been recently debated whether these preferences remain approximately constant during evolution or whether, due to coevolution of sites, they change strongly. Such research highlights an unresolved and fundamental issue with far-reaching implications for phylogenetic analysis and molecular evolution modeling. Here, we take advantage of the recent availability of phenotypically supported laboratory resurrections of Precambrian thioredoxins and β-lactamases to experimentally address the change of site-specific amino acid preferences over long geological timescales. Extensive mutational analyses support the notion that evolutionary adjustment to a new amino acid may occur, but to a large extent this is insufficient to erase the primitive preference for amino acid replacements. Generally, site-specific amino acid preferences appear to remain conserved throughout evolutionary history despite local sequence divergence. We show such preference conservation to be readily understandable in molecular terms and we provide crystallographic evidence for an intriguing structural-switch mechanism: Energetic preference for an ancestral amino acid in a modern protein can be linked to reorganization upon mutation to the ancestral local structure around the mutated site. Finally, we point out that site-specific preference conservation naturally leads to one plausible evolutionary explanation for the existence of intragenic global suppressor mutations.

  2. Mutational studies on resurrected ancestral proteins reveal conservation of site-specific amino acid preferences throughout evolutionary history.

    PubMed

    Risso, Valeria A; Manssour-Triedo, Fadia; Delgado-Delgado, Asunción; Arco, Rocio; Barroso-delJesus, Alicia; Ingles-Prieto, Alvaro; Godoy-Ruiz, Raquel; Gavira, Jose A; Gaucher, Eric A; Ibarra-Molero, Beatriz; Sanchez-Ruiz, Jose M

    2015-02-01

    Local protein interactions ("molecular context" effects) dictate amino acid replacements and can be described in terms of site-specific, energetic preferences for any different amino acid. It has been recently debated whether these preferences remain approximately constant during evolution or whether, due to coevolution of sites, they change strongly. Such research highlights an unresolved and fundamental issue with far-reaching implications for phylogenetic analysis and molecular evolution modeling. Here, we take advantage of the recent availability of phenotypically supported laboratory resurrections of Precambrian thioredoxins and β-lactamases to experimentally address the change of site-specific amino acid preferences over long geological timescales. Extensive mutational analyses support the notion that evolutionary adjustment to a new amino acid may occur, but to a large extent this is insufficient to erase the primitive preference for amino acid replacements. Generally, site-specific amino acid preferences appear to remain conserved throughout evolutionary history despite local sequence divergence. We show such preference conservation to be readily understandable in molecular terms and we provide crystallographic evidence for an intriguing structural-switch mechanism: Energetic preference for an ancestral amino acid in a modern protein can be linked to reorganization upon mutation to the ancestral local structure around the mutated site. Finally, we point out that site-specific preference conservation naturally leads to one plausible evolutionary explanation for the existence of intragenic global suppressor mutations. PMID:25392342

  3. A Comparison of Shear Bond Strengths of Metal and Ceramic Brackets using Conventional Acid Etching Technique and Er:YAG Laser Etching

    PubMed Central

    Yassaei, Sogra; Fekrazad, Reza; Shahraki, Neda; Goldani Moghadam, Mahdjoube

    2014-01-01

    Background and aims. The aim of this study was to compare shear bond strength (SBS) of metal and ceramic brackets bonded to enamel using acid versus Er:YAG laser etching. Materials and methods. Eighty premolars were divided into 4 groups: AM (acid etching/ metal brackets), AC (acid etching/ ceramic brackets), LM (laser etching/ metal brackets) and LC (laser etching/ ceramic brackets). Enamel condition-ing was done using acid in AC and AM and Er:YAG laser in LC and LM. Brackets were debonded with a Dartec machine and the SBSs were determined. Adhesive remnant index was evaluated under a stereomicroscope. Two additional teeth were conditioned with acid and laser for scanning electron microscopy examination. Comparisons of SBS value were done by ANOVA test. Results. statistical analyses showed that SBSs of acid groups were significantly higher than that of laser groups, but dif-ferences between SBS values of AC/ AM and LC/LM were not significant. SEM examination revealed different etching pattern. Conclusion. Low power Er:YAG laser etching offers clinically acceptable SBS which besides its other superiorities to acid etching can be an appropriate alternative for bonding of ceramic brackets. PMID:25024836

  4. Correlations Between Amino Acids at Different Sites in Local Sequences of Protein Fragments with Given Structural Patterns

    NASA Astrophysics Data System (ADS)

    Lu, Wen; Liu, Hai-yan

    2007-02-01

    Ample evidence suggests that the local structures of peptide fragments in native proteins are to some extent encoded by their local sequences. Detecting such local correlations is important but it is still an open question what would be the most appropriate method. This is partly because conventional sequence analyses treat amino acid preferences at each site of a protein sequence independently, while it is often the inter-site interactions that bring about local sequence-structure correlations. Here a new scheme is introduced to capture the correlation between amino acid preferences at different sites for different local structure types. A library of nine-residue fragments is constructed, and the fragments are divided into clusters based on their local structures. For each local structure cluster or type, chi-square tests are used to identify correlated preferences of amino acid combinations at pairs of sites. A score function is constructed including both the single site amino acid preferences and the dual-site amino acid combination preferences, which can be used to identify whether a sequence fragment would have a strong tendency to form a particular local structure in native proteins. The results show that, given a local structure pattern, dual-site amino acid combinations contain different information from single site amino acid preferences. Representative examples show that many of the statistically identified correlations agree with previously-proposed heuristic rules about local sequence-structure correlations, or are consistent with physical-chemical interactions required to stabilize particular local structures. Results also show that such dual-site correlations in the score function significantly improves the Z-score matching a sequence fragment to its native local structure relative to non-native local structures, and certain local structure types are highly predictable from the local sequence alone if inter-site correlations are considered.

  5. The mechanism of reduction of single-site redox proteins by ascorbic acid.

    PubMed Central

    Al-Ayash, A I; Wilson, M T

    1979-01-01

    The reduction of single-site haem and copper redox proteins by ascorbic acid was studied as a function of pH. Evidence is presented that indicates that the double-deprotonated ascorbate anion, ascorbate2-, is the reducing agent, and the pH-independent second-order rate constants for reduction by this species are given. Investigation of the temperature dependences of these rate constants have yielded the values of the activation parameters (delta H++ and delta S++) for reduction. These values, together with ligand-replacement studies, suggest that ascorbate2- acts as an outer-sphere reductant for these proteins. Reasons to account for the apparent inability of ascorbic acid to reduce the alkaline conformer of mammalian ferricytochrome c are suggested. PMID:35158

  6. REMOVING SLUDGE HEELS FROM SAVANNAH RIVER SITE WASTE TANKS BY OXALIC ACID DISSOLUTION

    SciTech Connect

    Poirier, M; David Herman, D; Fernando Fondeur, F; John Pareizs, J; Michael Hay, M; Bruce Wiersma, B; Kim Crapse, K; Thomas Peters, T; Samuel Fink, S; Donald Thaxton, D

    2009-03-01

    The Savannah River Site (SRS) will remove sludge as part of waste tank closure operations. Typically the bulk sludge is removed by mixing it with supernate to produce a slurry, and transporting the slurry to a downstream tank for processing. Experience shows that a residual heel may remain in the tank that cannot be removed by this conventional technique. In the past, SRS used oxalic acid solutions to disperse or dissolve the sludge heel to complete the waste removal. To better understand the actual conditions of oxalic acid cleaning of waste from carbon steel tanks, the authors developed and conducted an experimental program to determine its effectiveness in dissolving sludge, the hydrogen generation rate, the generation rate of other gases, the carbon steel corrosion rate, the impact of mixing on chemical cleaning, the impact of temperature, and the types of precipitates formed during the neutralization process. The test samples included actual SRS sludge and simulated SRS sludge. The authors performed the simulated waste tests at 25, 50, and 75 C by adding 8 wt % oxalic acid to the sludge over seven days. They conducted the actual waste tests at 50 and 75 C by adding 8 wt % oxalic acid to the sludge as a single batch. Following the testing, SRS conducted chemical cleaning with oxalic acid in two waste tanks. In Tank 5F, the oxalic acid (8 wt %) addition occurred over seven days, followed by inhibited water to ensure the tank contained enough liquid to operate the mixer pumps. The tank temperature during oxalic acid addition and dissolution was approximately 45 C. The authors analyzed samples from the chemical cleaning process and compared it with test data. The conclusions from the work are: (1) Oxalic acid addition proved effective in dissolving sludge heels in the simulant demonstration, the actual waste demonstration, and in SRS Tank 5F. (2) The oxalic acid dissolved {approx} 100% of the uranium, {approx} 100% of the iron, and {approx} 40% of the manganese

  7. Comparison of Shear Bond Strength of Orthodontic Brackets Bonded to Enamel Prepared By Er:YAG Laser and Conventional Acid-Etching

    PubMed Central

    Hosseini, M.H.; Namvar, F.; Chalipa, J.; Saber, K.; Chiniforush, N.; Sarmadi, S.; Mirhashemi, A.H.

    2012-01-01

    Introduction: The purpose of this study was to compare shear bond strength (SBS) of orthodontic brackets bonded to enamel prepared by Er:YAG laser with two different powers and conventional acid-etching. Materials and Methods: Forty-five human premolars extracted for orthodontic purposes were randomly assigned to three groups based on conditioning method: Group 1- conventional etching with 37% phosphoric acid; Group 2- irradiation with Er:YAG laser at 1 W; and Group 3- irradiation with Er:YAG laser at 1.5 W. Metal brackets were bonded on prepared enamel using a light-cured composite. All groups were subjected to thermocycling process. Then, the specimens mounted in auto-cure acryle and shear bond strength were measured using a universal testing machine with a crosshead speed of 0.5 mm per second. After debonding, the amount of resin remaining on the teeth was determined using the adhesive remnant index (ARI) scored 1 to 5. One-way analysis of variance was used to compare shear bond strengths and the Kruskal-Wallis test was performed to evaluate differences in the ARI for different etching types. Results: The mean and standard deviation of conventional acid-etch group, 1W laser group and 1.5W laser group was 3.82 ± 1.16, 6.97 ± 3.64 and 6.93 ± 4.87, respectively. Conclusion: The mean SBS obtained with an Er:YAG laser operated at 1W or 1.5W is approximately similar to that of conventional etching. However, the high variability of values in bond strength of irradiated enamel should be considered to find the appropriate parameters for applying Er:YAG laser as a favorable alternative for surface conditioning. PMID:22924098

  8. The species- and site-specific acid-base properties of penicillamine and its homodisulfide

    NASA Astrophysics Data System (ADS)

    Mirzahosseini, Arash; Szilvay, András; Noszál, Béla

    2014-08-01

    Penicillamine, penicillamine disulfide and 4 related compounds were studied by 1H NMR-pH titrations and case-tailored evaluation methods. The resulting acid-base properties are quantified in terms of 14 macroscopic and 28 microscopic protonation constants and the concomitant 7 interactivity parameters. The species- and site-specific basicities are interpreted by means of inductive and shielding effects through various intra- and intermolecular comparisons. The thiolate basicities determined this way are key parameters and exclusive means for the prediction of thiolate oxidizabilities and chelate forming properties in order to understand and influence chelation therapy and oxidative stress at the molecular level.

  9. Locations of the three primary binding sites for long-chain fatty acids on bovine serum albumin

    SciTech Connect

    Hamilton, J.A.; Era, S.; Bhamidipati, S.P. ); Reed, R.G. )

    1991-03-15

    Binding of {sup 13}C-enriched oleic acid to bovine serum albumin and to three large proteolytic fragments of albumin - two complementary fragments corresponding to the two halved of albumin and one fragment corresponding to the carboxyl-terminal domain - yielded unique patterns of NMR resonances (chemical shifts and relative intensities) that were used to identify the locations of binding of the first 5 mol of oleic acid to the multidomain albumin molecule. The first 3 mol of oleic acid added to intact albumin generated three distinct NMR resonances as a result of simultaneous binding of oleic acid to three heterogeneous sites (primary sites). This distribution suggests albumin to be a less symmetrical binding molecule than theoretical models predict. This work also demonstrates the power of NMR for the study of microenvironments of individual fatty acid binding sites in specific domain.

  10. The effect of the distance between acidic site and basic site immobilized on mesoporous solid on the activity in catalyzing aldol condensation

    SciTech Connect

    Yu Xiaofang; Yu Xiaobo; Wu Shujie; Liu Bo; Liu Heng; Guan Jingqi; Kan Qiubin

    2011-02-15

    Acid-base bifunctional heterogeneous catalysts containing carboxylic and amine groups, which were immobilized at defined distance from one another on the mesoporous solid were synthesized by immobilizing lysine onto carboxyl-SBA-15. The obtained materials were characterized by X-ray diffraction (XRD), N{sub 2} adsorption, Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron micrographs (SEM), transmission electron micrographs (TEM), elemental analysis, and back titration. Proximal-C-A-SBA-15 with a proximal acid-base distance was more active than maximum-C-A-SBA-15 with a maximum acid-base distance in aldol condensation reaction between acetone and various aldehydes. It appears that the distance between acidic site and basic site immobilized on mesoporous solid should be an essential factor for catalysis optimization. -- Graphical abstract: Proximal-C-A-SBA-15 with a proximal acid-base distance and maximum-C-A-SBA-15 with a maximum acid-base distance were synthesized by immobilizing lysine onto carboxyl-SBA-15. Display Omitted Research highlights: {yields} Proximal-C-A-SBA-15 with a proximal acid-base distance. {yields} Maximum-C-A-SBA-15 with a maximum acid-base distance. {yields} Compared to maximum-C-A-SBA-15, proximal-C-A-SBA-15 was more active toward aldol condensation reaction between acetone and various aldehydes.

  11. SAVANNAH RIVER SITE TANK CLEANING: CORROSION RATE FOR ONE VERSUS EIGHT PERCENT OXALIC ACID SOLUTION

    SciTech Connect

    Ketusky, E.; Subramanian, K.

    2011-01-20

    Until recently, the use of oxalic acid for chemically cleaning the Savannah River Site (SRS) radioactive waste tanks focused on using concentrated 4 and 8-wt% solutions. Recent testing and research on applicable dissolution mechanisms have concluded that under appropriate conditions, dilute solutions of oxalic acid (i.e., 1-wt%) may be more effective. Based on the need to maximize cleaning effectiveness, coupled with the need to minimize downstream impacts, SRS is now developing plans for using a 1-wt% oxalic acid solution. A technology gap associated with using a 1-wt% oxalic acid solution was a dearth of suitable corrosion data. Assuming oxalic acid's passivation of carbon steel was proportional to the free oxalate concentration, the general corrosion rate (CR) from a 1-wt% solution may not be bound by those from 8-wt%. Therefore, after developing the test strategy and plan, the corrosion testing was performed. Starting with the envisioned process specific baseline solvent, a 1-wt% oxalic acid solution, with sludge (limited to Purex type sludge-simulant for this initial effort) at 75 C and agitated, the corrosion rate (CR) was determined from the measured weight loss of the exposed coupon. Environmental variations tested were: (a) Inclusion of sludge in the test vessel or assuming a pure oxalic acid solution; (b) acid solution temperature maintained at 75 or 45 C; and (c) agitation of the acid solution or stagnant. Application of select electrochemical testing (EC) explored the impact of each variation on the passivation mechanisms and confirmed the CR. The 1-wt% results were then compared to those from the 8-wt%. The immersion coupons showed that the maximum time averaged CR for a 1-wt% solution with sludge was less than 25-mils/yr for all conditions. For an agitated 8-wt% solution with sludge, the maximum time averaged CR was about 30-mils/yr at 50 C, and 86-mils/yr at 75 C. Both the 1-wt% and the 8-wt% testing demonstrated that if the sludge was removed from

  12. Effect of cavity preparation method on microtensile bond strength of a self-etching primer vs phosphoric acid etchant to enamel.

    PubMed

    de Souza-Zaroni, Wanessa Christine; Delfino, Carina Sinclér; Ciccone-Nogueira, Juliane Cristina; Palma-Dibb, Regina Guenka; Corona, Silmara Aparecida Milori

    2007-10-01

    This study evaluated the effect of cavity preparation using air abrasion or carbide bur on bond strength to enamel treated with a self-etching primer (Tyrian SPE) or a phosphoric acid etchant. Twenty-four molars were divided into three groups: high-speed; standard handpiece (ST air abrasion) or supersonic handpiece (SP air abrasion) of the same air-abrasive system. The enamel surfaces were treated with one of the two etchants and the same adhesive agent One Step Plus, and then composite buildups were done with Filtek Z250. After 24 h at 37 degrees C, beams (0.8 mm2) were obtained and subjected to tensile stress in a universal testing machine (0.5 mm/min). The data were submitted to analysis of variance and Tukey's test (P < 0.05). For the conditioning agents, it was observed that the specimens conditioned with phosphoric acid presented superior results than the specimens that used Tyrian SPE. For the preparation techniques, it was verified that the SP air abrasion groups showed the highest bond strengths and carbide-bur groups presented the lowest bond strengths when the specimens were conditioned with Tyrian SPE. It can be concluded that the influence of the cavity preparation method was dependent on the conditioning system used, only when using carbide-bur preparation technique.

  13. Transport and deposition of Suwannee River Humic Acid/Natural Organic Matter formed silver nanoparticles on silica matrices: the influence of solution pH and ionic strength.

    PubMed

    Akaighe, Nelson; Depner, Sean W; Banerjee, Sarbajit; Sohn, Mary

    2013-07-01

    The transport and deposition of silver nanoparticles (AgNPs) formed from Ag(+) reduction by Suwannee River Humic Acid (SRHA) and Suwannee River Natural Organic Matter (SRNOM) utilizing a silica matrix is reported. The morphology and stability of the AgNPs was analyzed by transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential measurements. The percentage conversion of the initial [Ag(+)] to [AgNPs] was determined from a combination of atomic absorption (AAS) and UV-Vis spectroscopy, and centrifugation techniques. The results indicate higher AgNP transport and consequently low deposition in the porous media at basic pH conditions and low ionic strength. However, at low acidic pH and high ionic strength, especially with the divalent metallic cations, the mobility of the AgNPs in the porous media was very low, most likely due to NP aggregation. Overall, the results suggest the potential for AgNP contamination of subsurface soils and groundwater aquifers is mostly dependent on their aggregation state, controlled by the soil water and sediment ionic strength and pH.

  14. Promotion of viral internal ribosomal entry site-mediated translation under amino acid starvation.

    PubMed

    Licursi, Maria; Komatsu, Yumiko; Pongnopparat, Theerawat; Hirasawa, Kensuke

    2012-05-01

    Cap-dependent and internal ribosomal entry site (IRES)-mediated translation are regulated differently within cells. Viral IRES-mediated translation often remains active when cellular cap-dependent translation is severely impaired under cellular stresses induced by virus infection. To investigate how cellular stresses influence the efficiency of viral IRES-mediated translation, we used a bicistronic luciferase reporter construct harbouring IRES elements from the following viruses: encephalomyocarditis virus (EMCV), foot-and-mouth disease virus (FMDV), hepatitis C virus (HCV) or human rhinovirus (HRV). NIH3T3 cells transfected with these bicistronic reporter constructs were subjected to different cellular stresses. Increased translation initiation was only observed under amino acid starvation when EMCV or FMDV IRES elements were present. To identify cellular mechanisms that promoted viral IRES-mediated translation, we tested the involvement of eukaryotic initiation factor 4E-binding protein (4E-BP), general control non-depressed 2 (GCN2) and eukaryotic initiation factor 2B (eIF2B), as these are known to be modulated under amino acid starvation. Knockdown of 4E-BP1 impaired the promotion of EMCV and FMDV IRES-mediated translation under amino acid starvation, whereas GCN2 and eIF2B were not involved. To further investigate how 4E-BP1 regulates translation initiated by EMCV and FMDV IRES elements, we used a phosphoinositide kinase-3 inhibitor (LY294002), an mTOR inhibitor (Torin1) or leucine starvation to mimic 4E-BP1 dephosphorylation induced by amino acid starvation. 4E-BP1 dephosphorylation induced by the treatments was not sufficient to promote viral IRES-mediated translation. These results suggest that 4E-BP1 regulates EMCV and FMDV IRES-mediated translation under amino acid starvation, but not via its dephosphorylation. PMID:22302880

  15. Hatching success in salamanders and chorus frogs at two sites in Colorado, USA: Effects of acidic deposition and climate

    USGS Publications Warehouse

    Muths, E.; Campbell, D.H.; Corn, P.S.

    2003-01-01

    The snowpack in the vicinity of the Mount Zirkel Wilderness Area is among the most acidic in the western United States. We analyzed water chemistry and examined hatching success in tiger salamanders and chorus frogs at ponds there and at nearby Rabbit Ears Pass (Dumont) to determine whether acid deposition affects amphibians or their breeding habitats at these potentially sensitive locations. We found a wide range of acid neutralizing capacity among ponds within sites; the minimum pH recorded during the experiment was 5.4 at one of 12 ponds with all others at pH ??? 5.7. At Dumont, hatching success for chorus frogs was greater in ponds with low acid neutralizing capacity; however, lowest pHs were >5.8. At current levels of acid deposition, weather and pond characteristics are likely more important than acidity in influencing hatching success in amphibian larvae at these sites.

  16. State-of-the-Art pH Electrode Quality Control for Measurements of Acidic, Low Ionic Strength Waters.

    ERIC Educational Resources Information Center

    Stapanian, Martin A.; Metcalf, Richard C.

    1990-01-01

    Described is the derivation of the relationship between the pH measurement error and the resulting percentage error in hydrogen ion concentration including the use of variable activity coefficients. The relative influence of the ionic strength of the solution on the percentage error is shown. (CW)

  17. Amino acid sequence surrounding the chondroitin sulfate attachment site of thrombomodulin regulates chondroitin polymerization.

    PubMed

    Izumikawa, Tomomi; Kitagawa, Hiroshi

    2015-05-01

    Thrombomodulin (TM) is a cell-surface glycoprotein and a critical mediator of endothelial anticoagulant function. TM exists as both a chondroitin sulfate (CS) proteoglycan (PG) form and a non-PG form lacking a CS chain (α-TM); therefore, TM can be described as a part-time PG. Previously, we reported that α-TM bears an immature, truncated linkage tetrasaccharide structure (GlcAβ1-3Galβ1-3Galβ1-4Xyl). However, the biosynthetic mechanism to generate part-time PGs remains unclear. In this study, we used several mutants to demonstrate that the amino acid sequence surrounding the CS attachment site influences the efficiency of chondroitin polymerization. In particular, the presence of acidic residues surrounding the CS attachment site was indispensable for the elongation of CS. In addition, mutants defective in CS elongation did not exhibit anti-coagulant activity, as in the case with α-TM. Together, these data support a model for CS chain assembly in which specific core protein determinants are recognized by a key biosynthetic enzyme involved in chondroitin polymerization.

  18. Heterogeneous ceria catalyst with water-tolerant Lewis acidic sites for one-pot synthesis of 1,3-diols via Prins condensation and hydrolysis reactions.

    PubMed

    Wang, Yehong; Wang, Feng; Song, Qi; Xin, Qin; Xu, Shutao; Xu, Jie

    2013-01-30

    The use of a heterogeneous Lewis acid catalyst, which is insoluble and easily separable during the reaction, is a promising option for hydrolysis reactions from both environmental and practical viewpoints. In this study, ceria showed excellent catalytic activity in the hydrolysis of 4-methyl-1,3-dioxane to 1,3-butanediol in 95% yield and in the one-pot synthesis of 1,3-butanediol from propylene and formaldehyde via Prins condensation and hydrolysis reactions in an overall yield of 60%. In-depth investigations revealed that ceria is a water-tolerant Lewis acid catalyst, which has seldom been reported previously. The ceria catalysts showed rather unusual high activity in hydrolysis, with a turnover number (TON) of 260, which is rather high for bulk oxide catalysts, whose TONs are usually less than 100. Our conclusion that ceria functions as a Lewis acid catalyst in hydrolysis reactions is firmly supported by thorough characterizations with IR and Raman spectroscopy, acidity measurements with IR and (31)P magic-angle-spinning NMR spectroscopy, Na(+)/H(+) exchange tests, analyses using the in situ active-site capping method, and isotope-labeling studies. A relationship between surface vacancy sites and catalytic activity has been established. CeO(2)(111) has been confirmed to be the catalytically active crystalline facet for hydrolysis. Water has been found to be associatively adsorbed on oxygen vacancy sites with medium strength, which does not lead to water dissociation to form stable hydroxides. This explains why the ceria catalyst is water-tolerant. PMID:23228093

  19. Active-site mutations of diphtheria toxin: effects of replacing glutamic acid-148 with aspartic acid, glutamine, or serine.

    PubMed

    Wilson, B A; Reich, K A; Weinstein, B R; Collier, R J

    1990-09-18

    Glutamic acid-148, an active-site residue of diphtheria toxin identified by photoaffinity labeling with NAD, was replaced with aspartic acid, glutamine, or serine by directed mutagenesis of the F2 fragment of the toxin gene. Wild-type and mutant F2 proteins were synthesized in Escherichia coli, and the corresponding enzymic fragment A moieties (DTA) were derived, purified, and characterized. The Glu----Asp (E148D), Glu----Gln (E148Q), and Glu----Ser (E148S) mutations caused reductions in NAD:EF-2 ADP-ribosyltransferase activity of ca. 100-, 250-, and 300-fold, respectively, while causing only minimal changes in substrate affinity. The effects of the mutations on NAD-glycohydrolase activity were considerably different; only a 10-fold reduction in activity was observed for E148S, and the E148D and E148Q mutants actually exhibited a small but reproducible increase in NAD-glycohydrolytic activity. Photolabeling by nicotinamide-radiolabeled NAD was diminished ca. 8-fold in the E148D mutant and was undetectable in the other mutants. The results confirm that Glu-148 plays a crucial role in the ADP-ribosylation of EF-2 and imply an important function for the side-chain carboxyl group in catalysis. The carboxyl group is also important for photochemical labeling by NAD but not for NAD-glycohydrolase activity. The pH dependence of the catalytic parameters for the ADP-ribosyltransferase reaction revealed a group in DTA-wt that titrates with an apparent pKa of 6.2-6.3 and is in the protonated state in the rate-determining step.(ABSTRACT TRUNCATED AT 250 WORDS)

  20. Leaching of glyphosate and amino-methylphosphonic acid from Danish agricultural field sites.

    PubMed

    Kjaer, Jeanne; Olsen, Preben; Ullum, Marlene; Grant, Ruth

    2005-01-01

    Pesticide leaching is an important process with respect to contamination risk to the aquatic environment. The risk of leaching was thus evaluated for glyphosate (N-phosphonomethyl-glycine) and its degradation product AMPA (amino-methylphosphonic acid) under field conditions at one sandy and two loamy sites. Over a 2-yr period, tile-drainage water, ground water, and soil water were sampled and analyzed for pesticides. At a sandy site, the strong soil sorption capacity and lack of macropores seemed to prevent leaching of both glyphosate and AMPA. At one loamy site, which received low precipitation with little intensity, the residence time within the root zone seemed sufficient to prevent leaching of glyphosate, probably due to degradation and sorption. Minor leaching of AMPA was observed at this site, although the concentration was generally low, being on the order of 0.05 microg L(-1) or less. At another loamy site, however, glyphosate and AMPA leached from the root zone into the tile drains (1 m below ground surface [BGS]) in average concentrations exceeding 0.1 microg L(-1), which is the EU threshold value for drinking water. The leaching of glyphosate was mainly governed by pronounced macropore flow occurring within the first months after application. AMPA was frequently detected more than 1.5 yr after application, thus indicating a minor release and limited degradation capacity within the soil. Leaching has so far been confined to the depth of the tile drains, and the pesticides have rarely been detected in monitoring screens located at lower depths. This study suggests that as both glyphosate and AMPA can leach through structured soils, they thereby pose a potential risk to the aquatic environment.

  1. Composite active site of chondroitin lyase ABC accepting both epimers of uronic acid

    SciTech Connect

    Shaya, D.; Hahn, Bum-Soo; Bjerkan, Tonje Marita; Kim, Wan Seok; Park, Nam Young; Sim, Joon-Soo; Kim, Yeong-Shik; Cygler, M.

    2008-03-19

    Enzymes have evolved as catalysts with high degrees of stereospecificity. When both enantiomers are biologically important, enzymes with two different folds usually catalyze reactions with the individual enantiomers. In rare cases a single enzyme can process both enantiomers efficiently, but no molecular basis for such catalysis has been established. The family of bacterial chondroitin lyases ABC comprises such enzymes. They can degrade both chondroitin sulfate (CS) and dermatan sulfate (DS) glycosaminoglycans at the nonreducing end of either glucuronic acid (CS) or its epimer iduronic acid (DS) by a {beta}-elimination mechanism, which commences with the removal of the C-5 proton from the uronic acid. Two other structural folds evolved to perform these reactions in an epimer-specific fashion: ({alpha}/{alpha}){sub 5} for CS (chondroitin lyases AC) and {beta}-helix for DS (chondroitin lyases B); their catalytic mechanisms have been established at the molecular level. The structure of chondroitinase ABC from Proteus vulgaris showed surprising similarity to chondroitinase AC, including the presence of a Tyr-His-Glu-Arg catalytic tetrad, which provided a possible mechanism for CS degradation but not for DS degradation. We determined the structure of a distantly related Bacteroides thetaiotaomicron chondroitinase ABC to identify additional structurally conserved residues potentially involved in catalysis. We found a conserved cluster located {approx}12 {angstrom} from the catalytic tetrad. We demonstrate that a histidine in this cluster is essential for catalysis of DS but not CS. The enzyme utilizes a single substrate-binding site while having two partially overlapping active sites catalyzing the respective reactions. The spatial separation of the two sets of residues suggests a substrate-induced conformational change that brings all catalytically essential residues close together.

  2. Evaluation Of Sludge Heel Dissolution Efficiency With Oxalic Acid Cleaning At Savannah River Site

    SciTech Connect

    Sudduth, Christie; Vitali, Jason; Keefer, Mark

    2014-01-08

    The chemical cleaning process baseline strategy at the Savannah River Site was revised to improve efficiency during future execution of the process based on lessons learned during previous bulk oxalic acid cleaning activities and to account for operational constraints imposed by safety basis requirements. These improvements were also intended to transcend the difficulties that arise from waste removal in higher rheological yield stress sludge tanks. Tank 12 implemented this improved strategy and the bulk oxalic acid cleaning efforts concluded in July 2013. The Tank 12 radiological removal results were similar to previous bulk oxalic acid cleaning campaigns despite the fact that Tank 12 contained higher rheological yield stress sludge that would make removal more difficult than the sludge treated in previous cleaning campaigns. No appreciable oxalate precipitation occurred during the cleaning process in Tank 12 compared to previous campaigns, which aided in the net volume reduction of 75-80%. Overall, the controls established for Tank 12 provide a template for an improved cleaning process.

  3. Varying ratios of omega-6: omega-3 fatty acids on the pre-and postmortem bone mineral density, bone ash, and bone breaking strength of laying chickens.

    PubMed

    Baird, H T; Eggett, D L; Fullmer, S

    2008-02-01

    The purpose of this study was to investigate the effects of varying ratios of n-6 to n-3 fatty acids in the diets of White Leghorn chickens on tibia bone characteristics [bone mineral density, bone mineral content (BMC), ash bone mineral content, bone morphology, and cortical thickness] and tibia bone strength parameters (ultimate force, bending stress, maximum strain, Young's modulus of elasticity, area under the curve, and moment of inertia). Seventy-five 16-wk-old female White Leghorn chickens were randomly assigned to 1 of 5 dietary ratios of n-6 to n-3 fatty acids: 47.8:1, 18.0:1, 7.6:1, 5.9:1, or 4.7:1. Corn oil was the n-6 fatty acid source, whereas flax oil provided the n-3 fatty acids. Bone density was measured on the left tibia via dual-energy x-ray absorptiometry (DXA) prior to killing and after excision. Bones were ashed in a muffle furnace at 500 degrees F. Tibia bones were broken by using a 3-point bending rig. Results showed no significant effect of diet on bone characteristics. There were no significant differences among diet groups for parameters of bone strength except cortical thickness (P < or = 0.01). Bone mineral content determined by ashing was significantly different by 9.2% (P < or = 0.0001) from BMC determined in vivo by DXA; however, there were no differences in ex vivo BMC and BMC ash, although they were highly correlated (r = 0.99, P < or = 0.0001). We concluded that there was no effect of n-3 fatty acids on tibia bone in mature White Leghorn chickens. The GE Lunar Prodigy DXA instrument significantly underestimated the in vivo BMC in chickens.

  4. Varying ratios of omega-6: omega-3 fatty acids on the pre-and postmortem bone mineral density, bone ash, and bone breaking strength of laying chickens.

    PubMed

    Baird, H T; Eggett, D L; Fullmer, S

    2008-02-01

    The purpose of this study was to investigate the effects of varying ratios of n-6 to n-3 fatty acids in the diets of White Leghorn chickens on tibia bone characteristics [bone mineral density, bone mineral content (BMC), ash bone mineral content, bone morphology, and cortical thickness] and tibia bone strength parameters (ultimate force, bending stress, maximum strain, Young's modulus of elasticity, area under the curve, and moment of inertia). Seventy-five 16-wk-old female White Leghorn chickens were randomly assigned to 1 of 5 dietary ratios of n-6 to n-3 fatty acids: 47.8:1, 18.0:1, 7.6:1, 5.9:1, or 4.7:1. Corn oil was the n-6 fatty acid source, whereas flax oil provided the n-3 fatty acids. Bone density was measured on the left tibia via dual-energy x-ray absorptiometry (DXA) prior to killing and after excision. Bones were ashed in a muffle furnace at 500 degrees F. Tibia bones were broken by using a 3-point bending rig. Results showed no significant effect of diet on bone characteristics. There were no significant differences among diet groups for parameters of bone strength except cortical thickness (P < or = 0.01). Bone mineral content determined by ashing was significantly different by 9.2% (P < or = 0.0001) from BMC determined in vivo by DXA; however, there were no differences in ex vivo BMC and BMC ash, although they were highly correlated (r = 0.99, P < or = 0.0001). We concluded that there was no effect of n-3 fatty acids on tibia bone in mature White Leghorn chickens. The GE Lunar Prodigy DXA instrument significantly underestimated the in vivo BMC in chickens. PMID:18212376

  5. Characteristics of fluoride in pore-water at accidental hydrofluoric acid spillage site, Gumi, Korea

    NASA Astrophysics Data System (ADS)

    Kwon, E. H.; Lee, H. A.; Lee, J.; Kim, D.; Lee, S.; Yoon, H. O.

    2015-12-01

    A leakage accident of hydrofluoric acid (HF) occurred in Gumi, South Korea at Sep. 2012. The study site is located in the borderline between a large-scale industrial complex and a rural area. The HF plume was made immediately, and moved toward the rural area through air. After the accident, 212 ha of farm land were influenced and most of crops were withered. To recover the soil, CaO was applied after six months. Although several studies have done to estimate the extension and movement of HF plume in the air and to assess the impact on human health or plant after the incident, the long-term fate of fluoride (F) in the affected soils is not identified clearly. Thus, this study aimed to understand the behavior of F in the soil after HF releasing from accident site through chemical analysis and geochemical modeling. Within the radius of 1 km of accident site, 16 pore-water and soil samples were collected. The semi-quantitative soil composition (i.e., Al, Ca, Fe, K, Mg, Mn, Na, P, Si, Ti), total F, total P, OM contents in soil, and soil pH have already been measured, and pore-water compositions are also identified. From these experimental and modeling data, we could be evaluate if impact of accident exists until now, and also could be select and identify existing form of fluoride in soil and pore-water.

  6. Will reduced sulphur emissions under the Second Sulphur Protocol lead to recovery of acid sensitive sites in UK?

    PubMed

    Jenkins, A; Helliwell, R C; Swingewood, P J; Sefton, C; Renshaw, M; Ferrier, R C

    1998-01-01

    A conceptual model of the combined effects of acid deposition and land-use, Model of Acidification of Groundwater In Catchments (MAGIC), was applied to 21 upland sites in the UK Acid Waters Monitoring Network (AWMN) to assess the likely future recovery in response to the latest international agreements controlling anthropogenic sulphur emissions throughout Europe. Future estimates of sulphur deposition were generated by the Hull Acid Rain Model (HARM), based on the agreed reductions outlined in the Second Sulphur Protocol. The results indicate only a limited degree of recovery in surface-water chemistry at all sites over the next 50 years; moreover, a continuing decline in soil base status is predicted to occur at 70% of sites, resulting in longer term reacidification of surface-water at 38% of sites. However, compared with a 'business as usual' scenario the recovery is pronounced, although acidified sites will require further reductions in acidic deposition if recovery to pre-industrial chemical conditions are to be achieved. Furthermore, land-use scenarios at afforested sites suggest that replanting of felled forest will lead to a further increase in acidification. This strengthens the argument that plantation forestry should be avoided in areas considered geologically sensitive to acidic deposition.

  7. Eight Weeks of Phosphatidic Acid Supplementation in Conjunction with Resistance Training Does Not Differentially Affect Body Composition and Muscle Strength in Resistance-Trained Men

    PubMed Central

    Andre, Thomas L.; Gann, Joshua J.; McKinley-Barnard, Sarah K.; Song, Joon J.; Willoughby, Darryn S.

    2016-01-01

    This study attempted to determine the effects of eight weeks of resistance training (RT) combined with phosphatidic acid (PA) supplementation at a dose of either 250 mg or 375 mg on body composition and muscle size and strength. Twenty-eight resistance-trained men were randomly assigned to ingest 375 mg [PA375 (n = 9)] or 250 mg [PA250 (n = 9)] of PA or 375 mg of placebo [PLC (n = 10)] daily for eight weeks with RT. Supplements were ingested 60 minutes prior to RT and in the morning on non-RT days. Participants’ body composition, muscle size, and lower-body muscle strength were determined before and after training/supplementation. Separate group x time ANOVAs for each criterion variable were used employing an alpha level of ≤ 0.05. Magnitude- based inferences were utilized to determine the likely or unlikely impact of PA on each criterion variable. A significant main effect for time was observed for improvements in total body mass (p = 0.003), lean mass (p = 0.008), rectus femoris cross-sectional area [RF CSA (p = 0.011)], and lower-body strength (p < 0.001), but no significant interactions were present (p > 0.05). Collectively, magnitude-based inferences determined both doses of PA to have a likely impact of increasing body mass (74.2%), lean mass (71.3%), RF CSA (92.2%), and very likely impact on increasing lower-body strength (98.1% beneficial). When combined with RT, it appears that PA has a more than likely impact on improving lower-body strength, whereas a likely impact exists for increasing muscle size and lean mass. Key points In response to eight weeks resistance training and PLC and PA (375 mg and 250 mg) supplementation, similar increases in lower-body muscle strength occurred in all three groups; however, the increases were not different between supplement groups. In response to eight weeks resistance training and PLC and PA (375 mg and 250 mg) supplementation, similar increases in lean mass occurred in all three groups; however, the increases were

  8. Amino acid conditions near the GPI anchor attachment site of prion protein for the conversion and the GPI anchoring.

    PubMed

    Hizume, Masaki; Kobayashi, Atsushi; Mizusawa, Hidehiro; Kitamoto, Tetsuyuki

    2010-01-22

    Prion protein (PrP) is a glycosylphosphatidylinositol (GPI)-anchored protein, and the C-terminal GPI anchor signal sequence (GPI-SS) of PrP is cleaved before GPI anchoring. However, mutations near the GPI anchor attachment site (the omega site) in the GPI-SS have been recognized in human genetic prion diseases. Moreover, the omega site of PrP has not been identified except hamster, though it is known that amino acid restrictions are very severe at the omega and omega+2 sites in other GPI-anchored proteins. To investigate the effect of mutations near the omega site of PrP on the conversion and the GPI anchoring, and to discover the omega site of murine PrP, we systematically created mutant murine PrP with all possible single amino acid substitutions at every amino acid residue from codon 228 to 240. We transfected them into scrapie-infected mouse neuroblastoma cells and examined the conversion efficiencies and the GPI anchoring of each mutant PrP. Mutations near the omega site altered the conversion efficiencies and the GPI anchoring efficiencies. Especially, amino acid restrictions for the conversion and the GPI anchoring were severe at codons 230 and 232 in murine PrP, though they were less severe than in other GPI-anchored proteins. Only the mutant PrPs presented on a cell surface via a GPI anchor were conversion competent. The present study shows that mutations in the GPI-SS can affect the GPI anchoring and the conversion efficiency of PrP. We clarified for the first time the omega site of murine PrP and the amino acid conditions near the omega site for the conversion as well as GPI anchoring.

  9. In-situ stress and strength in the Nankai inner accretionary prism at Site C0002, IODP NanTroSEIZE

    NASA Astrophysics Data System (ADS)

    Kitajima, H.; Valdez, R. D.; Kitamura, M.; Sone, H.; Saffer, D. M.; Tobin, H. J.; Hirose, T.; Kuo, S. T.

    2015-12-01

    As a part of the International Ocean Discovery Program (IODP) Nankai Trough Seismogenic Zone Experiment (NanTroSEIZE), a deep riser borehole has been drilled into the Kumano forearc basin and the underlying inner accretionary wedge at Site C0002, located ~35 km landward from the trench. One of the primary objectives of drilling the riser site was to characterize in-situ stress and pore pressure in the hanging wall above the locked plate boundary. Here, we: (1) investigate the mechanical strength and deformation behavior of prism sediment via laboratory experiments on core samples; and (2) quantify in-situ stress (Sv, Sh, and SH), and pore pressure (Pp) in the Kumano basin and the inner prism. We conducted triaxial compression experiments on core samples recovered from ~ 2200 meters below sea floor (mbsf) during IODP Expedition 348, at effective pressures (Pe) ranging from 8 and 36 MPa, and at temperatures of either 25°C or 60°C. Our preliminary results indicate that the prism (20 - 42% porosity) rocks deform brittlely at Pe < 22 MPa, but exhibit strain hardening at Pe = 36 MPa. This pressure-porosity condition for a brittle-ductile transition is consistent with previous work defining yield models for incoming sediments at the Nankai Trough (Kitajima and Saffer, 2012). Combining P-wave velocity logs and downhole measurements of leak-off pressure at Site C0002 with an empirical relationship between P-wave velocity, porosity, and effective stress, we show that the Kumano forearc basin is in a uniaxial-strain loading path, which defines a normal faulting stress regime (Sv>SH>=Sh), whereas the inner accretionary prism is in a triaxial-strain loading path that defines a strike-slip faulting regime (SH>Sv>Sh). We estimate excess pore pressure below ~2000 mbsf ranging from 0-12 MPa, corresponding to a pore pressure ratio λ* of 0 - 0.40.

  10. The species- and site-specific acid-base properties of biological thiols and their homodisulfides.

    PubMed

    Mirzahosseini, Arash; Noszál, Béla

    2014-07-01

    Cysteamine, cysteine, homocysteine, their homodisulfides and 9 related compounds were studied by ¹H NMR-pH titrations and case-tailored evaluation methods. The resulting acid-base properties are quantified in terms of 33 macroscopic and 62 microscopic protonation constants and the concomitant 16 interactivity parameters, providing thus the first complete microspeciation of this vitally important family of biomolecules. The species- and site-specific basicities are interpreted by means of inductive and hydrogen-bonding effects through various intra- and intermolecular comparisons. The pH-dependent distribution of the microspecies is depicted. The thiolate basicities determined this way provide exclusive means for the prediction of thiolate oxidizabilities, a key parameter to understand and influence oxidative stress at the molecular level.

  11. 36C1 measurements and the hydrology of an acid injection site

    USGS Publications Warehouse

    Vourvopoulos, G.; Brahana, J.V.; Nolte, E.; Korschinek, G.; Priller, A.; Dockhorn, B.

    1990-01-01

    In an area in western Tennessee (United States), an industrial firm is injecting acidic (pH = 0.1) iron chloride into permeable zones of carbonate rocks at depths ranging from 1000 to 2200 m below land surface. Overlying the injection zone at a depth of approximately 500 m below land surface is a regional fresh-water aquifer, the Knox aquifer. A study is currently underway to investigate whether the injection wells are hydraulically isolated from the fresh-water aquifer. Drilling of a test well that will reach a total depth of 2700 m has been initiated. The 36Cl content of 15 samples from the Knox aquifer, from monitor wells in the vicinity of the injection site, and from the test well have been analyzed. ?? 1990.

  12. Application of a depositional facies model to an acid mine drainage site.

    PubMed

    Brown, Juliana F; Jones, Daniel S; Mills, Daniel B; Macalady, Jennifer L; Burgos, William D

    2011-01-01

    Lower Red Eyes is an acid mine drainage site in Pennsylvania where low-pH Fe(II) oxidation has created a large, terraced iron mound downstream of an anoxic, acidic, metal-rich spring. Aqueous chemistry, mineral precipitates, microbial communities, and laboratory-based Fe(II) oxidation rates for this site were analyzed in the context of a depositional facies model. Depositional facies were defined as pools, terraces, or microterracettes based on cm-scale sediment morphology, irrespective of the distance downstream from the spring. The sediments were composed entirely of Fe precipitates and cemented organic matter. The Fe precipitates were identified as schwertmannite at all locations, regardless of facies. Microbial composition was studied with fluorescence in situ hybridization (FISH) and transitioned from a microaerophilic, Euglena-dominated community at the spring, to a Betaproteobacteria (primarily Ferrovum spp.)-dominated community at the upstream end of the iron mound, to a Gammaproteobacteria (primarily Acidithiobacillus)-dominated community at the downstream end of the iron mound. Microbial community structure was more strongly correlated with pH and geochemical conditions than depositional facies. Intact pieces of terrace and pool sediments from upstream and downstream locations were used in flowthrough laboratory reactors to measure the rate and extent of low-pH Fe(II) oxidation. No change in Fe(II) concentration was observed with (60)Co-irradiated sediments or with no-sediment controls, indicating that abiotic Fe(II) oxidation was negligible. Upstream sediments attained lower effluent Fe(II) concentrations compared to downstream sediments, regardless of depositional facies.

  13. Gas-phase acidities of o-, m- and p-dehydrobenzoic acid radicals. Determination of the substituent constants for a phenyl radical site

    NASA Astrophysics Data System (ADS)

    Wenthold, Paul G.; Squires, Robert R.

    1998-05-01

    Reaction of CO2 with o-, m-, and p-benzyne radical anions in the gas phase produces o-, m-, and p-dehydrobenzoate radical anions, respectively. The (oxygen) gas-phase basicities of these ions, which are equivalent to the gas-phase acidities of the corresponding dehydrobenzoic acid radicals, [Delta]Gacid(o-, m-, or p-C6H4CO2-H), have been determined with a flowing afterglowtriple quadrupole apparatus by means of the kinetic method. The measured values are (in kcal mol-1): [Delta]Gacid(o-C6H4CO2H) = 330.4 +/- 0.4, [Delta]Gacid(m-C6H4CO2H) = 330.2 +/- 0.4, and [Delta]Gacid(p-C6H4CO2H) = 331.6 +/- 0.4 kcal mol-1. All three radicals are more acidic than benzoic acid ([Delta]Gacid = 333.1 +/- 2.0 kcal mol-1). The measured gas-phase acidities for the meta and para isomers suggest values for the resonance-effect substituent constant, [sigma]R, and the field/inductive effect substituent constant, [sigma]F, for a phenyl radical site of - 0.47 and 0.57, respectively. This classifies a phenyl radical site as a strong inductive withdrawing, and strong resonance donating substituent. Density functional calculations of the gas-phase acidities of dehydrobenzoic acids are in good agreement with the experimental results. The increased acidities of the dehydrobenzoic acids are shown to arise from a balance between the electron withdrawing effect of the electronegative radical site, and a compensating polarization of the [pi] system which mimics the effect of a resonance donor group located at the radical carbon.

  14. Activation of the Human Epithelial Sodium Channel (ENaC) by Bile Acids Involves the Degenerin Site.

    PubMed

    Ilyaskin, Alexandr V; Diakov, Alexei; Korbmacher, Christoph; Haerteis, Silke

    2016-09-16

    The epithelial sodium channel (ENaC) is a member of the ENaC/degenerin ion channel family, which also includes the bile acid-sensitive ion channel (BASIC). So far little is known about the effects of bile acids on ENaC function. ENaC is probably a heterotrimer consisting of three well characterized subunits (αβγ). In humans, but not in mice and rats, an additional δ-subunit exists. The aim of this study was to investigate the effects of chenodeoxycholic, cholic, and deoxycholic acid in unconjugated (CDCA, CA, and DCA) and tauro-conjugated (t-CDCA, t-CA, t-DCA) form on human ENaC in its αβγ- and δβγ-configuration. We demonstrated that tauro-conjugated bile acids significantly stimulate ENaC in the αβγ- and in the δβγ-configuration. In contrast, non-conjugated bile acids have a robust stimulatory effect only on δβγENaC. Bile acids stimulate ENaC-mediated currents by increasing the open probability of active channels without recruiting additional near-silent channels known to be activated by proteases. Stimulation of ENaC activity by bile acids is accompanied by a significant reduction of the single-channel current amplitude, indicating an interaction of bile acids with a region close to the channel pore. Analysis of the known ASIC1 (acid-sensing ion channel) crystal structure suggested that bile acids may bind to the pore region at the degenerin site of ENaC. Substitution of a single amino acid residue within the degenerin region of βENaC (N521C or N521A) significantly reduced the stimulatory effect of bile acids on ENaC, suggesting that this site is critical for the functional interaction of bile acids with the channel. PMID:27489102

  15. Site-specific acid-base properties of pholcodine and related compounds.

    PubMed

    Kovács, Z; Hosztafi, S; Noszál, B

    2006-11-01

    The acid-base properties of pholcodine, a cough-depressant agent, and related compounds including metabolites were studied by 1H NMR-pH titrations, and are characterised in terms of macroscopic and microscopic protonation constants. New N-methylated derivatives were also synthesized in order to quantitate site- and nucleus-specific protonation shifts and to unravel microscopic acid-base equilibria. The piperidine nitrogen was found to be 38 and 400 times more basic than its morpholine counterpart in pholcodine and norpholcodine, respectively. The protonation data show that the molecule of pholcodine bears an average of positive charge of 1.07 at physiological pH, preventing it from entering the central nervous system, a plausible reason for its lack of analgesic or addictive properties. The protonation constants of pholcodine and its derivatives are interpreted by comparing with related molecules of pharmaceutical interest. The pH-dependent relative concentrations of the variously protonated forms of pholcodine and morphine are depicted in distribution diagrams.

  16. Site-specific acid-base properties of pholcodine and related compounds.

    PubMed

    Kovács, Z; Hosztafi, S; Noszál, B

    2006-11-01

    The acid-base properties of pholcodine, a cough-depressant agent, and related compounds including metabolites were studied by 1H NMR-pH titrations, and are characterised in terms of macroscopic and microscopic protonation constants. New N-methylated derivatives were also synthesized in order to quantitate site- and nucleus-specific protonation shifts and to unravel microscopic acid-base equilibria. The piperidine nitrogen was found to be 38 and 400 times more basic than its morpholine counterpart in pholcodine and norpholcodine, respectively. The protonation data show that the molecule of pholcodine bears an average of positive charge of 1.07 at physiological pH, preventing it from entering the central nervous system, a plausible reason for its lack of analgesic or addictive properties. The protonation constants of pholcodine and its derivatives are interpreted by comparing with related molecules of pharmaceutical interest. The pH-dependent relative concentrations of the variously protonated forms of pholcodine and morphine are depicted in distribution diagrams. PMID:17004059

  17. Diversity of acidophilic prokaryotes at two acid mine drainage sites in Turkey.

    PubMed

    Aytar, Pınar; Kay, Catherine Melanie; Mutlu, Mehmet Burçin; Çabuk, Ahmet; Johnson, David Barrie

    2015-04-01

    The biodiversity of acidophilic prokaryotes in two acidic (pH 2.8-3.05) mine drainage (AMD) sites (Balya and Çan) in Turkey was examined using a combined cultivation-based and cultivation-independent approach. The latter included analyzing microbial diversity using fluorescent in situ hybridization (FISH), terminal restriction enzyme fragment length polymorphism (`T-RFLP), and quantitative PCR (qPCR). Numbers of cultivatable heterotrophic acidophilic bacteria were over an order of magnitude greater than those of chemolithotrophic acidophiles in both AMD ponds examined. Isolates identified as strains of Acidithiobacillus ferrivorans, Acidiphilium organovorum, and Ferrimicrobium acidiphilum were isolated from the Balya AMD pond, and others identified as strains of Leptospirillum ferriphilum, Acidicapsa ligni, and Acidiphilium rubrum from Çan AMD. Other isolates were too distantly related (from analysis of their 16S rRNA genes) to be identified at the species level. Archaeal diversity in the two ponds appeared to be far more limited. T-RFLP and qPCR confirmed the presence of Ferroplasma-like prokaryotes, but no archaea were isolated from the two sites. qPCR generated semiquantitative data for genera of some of the iron-oxidizing acidophiles isolated and/or detected, suggesting the order of abundance was Leptospirillum > Ferroplasma > Acidithiobacillus (Balya AMD) and Ferroplasma > Leptospirillum > Acidithiobacillus (Çan AMD).

  18. The Protonation Site of para-Dimethylaminobenzoic Acid Using Atmospheric Pressure Ionization Methods

    NASA Astrophysics Data System (ADS)

    Chai, Yunfeng; Weng, Guofeng; Shen, Shanshan; Sun, Cuirong; Pan, Yuanjiang

    2015-04-01

    The protonation site of para-dimethylaminobenzoic acid ( p-DMABA) was investigated using atmospheric pressure ionization methods (ESI and APCI) coupled with collision-induced dissociation (CID), nuclear magnetic resonance (NMR), and computational chemistry. Theoretical calculations and NMR experiments indicate that the dimethyl amino group is the preferred site of protonation both in the gas phase and aqueous solution. Protonation of p-DMABA occurs at the nitrogen atom by ESI independent of the solvents and other operation conditions under typical thermodynamic control. However, APCI produces a mixture of the nitrogen- and carbonyl oxygen-protonated p-DMABA when aprotic organic solvents (acetonitrile, acetone, and tetrahydrofuran) are used, exhibiting evident kinetic characteristics of protonation. But using protic organic solvents (methanol, ethanol, and isopropanol) in APCI still leads to the formation of thermodynamically stable N-protonated p-DMABA. These structural assignments were based on the different CID behavior of the N- and O-protonated p-DMABA. The losses of methyl radical and water are the diagnostic fragmentations of the N- and O-protonated p-DMABA, respectively. In addition, the N-protonated p-DMABA is more stable than the O-protonated p-DMABA in CID revealed by energy resolved experiments and theoretical calculations.

  19. Studies on the site of biosynthesis of acidic glycoproteins of guinea-pig serum

    PubMed Central

    Simkin, J. L.; Jamieson, J. C.

    1967-01-01

    1. Studies were carried out to determine the cellular and subcellular site of biosynthesis of components of fraction I, an α-globulin fraction containing acidic glycoproteins isolated from guinea-pig serum. l-[U-14C]Leucine or -valine and d-[1-14C]glucosamine were used as precursors. 2. A lag of about 10min. occurred before appreciable label appeared in fraction I of serum after injection of leucine or glucosamine. Label in fraction I after 60min. labelling with glucosamine was present almost entirely in hexosamine and sialic acid. 3. Site of synthesis was investigated by studies in vivo up to 17min. after injection of precursor. Particulate subcellular fractions isolated from liver, spleen and kidney or homogenates of the latter two tissues were extracted with Lubrol. Extracts were allowed to react by double diffusion with antisera to fraction I or to subfractions isolated from it, and gels were subsequently subjected to radioautography. With either amino acid or glucosamine as precursor, only extracts of the microsome fraction of liver formed precipitin lines that were appreciably radioactive. 4. The role of the microsome fraction of liver in the synthesis of these glycoproteins was confirmed by immunological studies after incubation of liver slices with leucine or glucosamine. Incorporation of leucine was also investigated in a cell-free microsome system. 5. Material was also precipitated from certain Lubrol extracts of liver microsomes by direct addition of antiserum and its radioactivity measured. Degradation of material thus precipitated and use of heterologous immune systems showed that labelling of precipitin lines represented biosynthesis. 6. A study of extraction procedures suggested that the substances present in the microsome fraction of liver that react with specific antisera are associated with membranous structures. 7. Most or all precipitin lines formed by Lubrol extracts of liver microsomes interacted with precipitin lines given by guinea

  20. Effects of amino acid sequence, buffers, and ionic strength on the rate and mechanism of deamidation of asparagine residues in small peptides.

    PubMed

    Tyler-Cross, R; Schirch, V

    1991-11-25

    The nonenzymatic rates of deamidation of Asn residues in a series of pentapeptides with the sequences VSNXV and VXNSV, where X is one of 10 different amino acids, were determined at neutral, alkaline, and acid pH values. The results demonstrate that in neutral and alkaline solutions the amino acid residue on the amino side of the Asn had little or no effect on the rate of deamidation regardless of its charge or size. The group on the carboxyl side of Asn affected the rate of deamidation significantly. Increasing size and branching in the side chain of this residue decreased the rate of deamidation by as much as 70-fold compared to glycine in the N-G sequence, which had the greatest rate of deamidation. In acidic solution, the rate of deamidation of the Asn residue was not affected by the amino acid sequence of the peptide. The products for each deamidation reaction were tested for the formation of isoAsp residues. In neutral and alkaline solutions, all products showed that the isoAsp:Asp peptide products were formed in about a 3:1 ratio. In acidic solution, the Asp peptide was the only deamidation product formed. All peptides in which a Ser residue follows the Asn residue were found to undergo a peptide cleavage reaction in neutral and alkaline solutions, yielding a tripeptide and a dipeptide. The rate of the cleavage reaction was about 10% of the rate of the deamidation pathway at neutral and alkaline pH values. The rates of deamidation of Asn residues in the peptides studied were not affected by ionic strength, and were not specific base catalyzed. General base catalysis was observed for small bases like ammonia. A model for the deamidation reaction is proposed to account for the observed effects. PMID:1939272

  1. Differing effects of PTH 1-34, PTH 1-84, and zoledronic acid on bone microarchitecture and estimated strength in postmenopausal women with osteoporosis: an 18-month open-labeled observational study using HR-pQCT.

    PubMed

    Hansen, Stinus; Hauge, Ellen M; Beck Jensen, Jens-Erik; Brixen, Kim

    2013-04-01

    Whereas the beneficial effects of intermittent treatment with parathyroid hormone (PTH) (intact PTH 1-84 or fragment PTH 1-34, teriparatide) on vertebral strength is well documented, treatment may not be equally effective in the peripheral skeleton. We used high-resolution peripheral quantitative computed tomography (HR-pQCT) to detail effects on compartmental geometry, density, and microarchitecture as well as finite element (FE) estimated integral strength at the distal radius and tibia in postmenopausal osteoporotic women treated with PTH 1-34 (20 µg sc daily, n = 18) or PTH 1-84 (100 µg sc daily, n = 20) for 18 months in an open-label, nonrandomized study. A group of postmenopausal osteoporotic women receiving zoledronic acid (5 mg infusion once yearly, n = 33) was also included. Anabolic therapy increased cortical porosity in radius (PTH 1-34 32 ± 37%, PTH 1-84 39 ± 32%, both p < 0.001) and tibia (PTH 1-34 13 ± 27%, PTH 1-84 15 ± 22%, both p < 0.001) with corresponding declines in cortical density. With PTH 1-34, increases in cortical thickness in radius (2.0 ± 3.8%, p < 0.05) and tibia (3.8 ± 10.4%, p < 0.01) were found. Trabecular number increased in tibia with both PTH 1-34 (4.2 ± 7.1%, p < 0.05) and PTH 1-84 (5.3 ± 8.3%, p < 0.01). Zoledronic acid did not impact cortical porosity at either site but increased cortical thickness (3.0 ± 3.5%, p < 0.01), total (2.7 ± 2.5%, p < 0.001) and cortical density (1.5 ± 2.0%, p < 0.01) in tibia as well as trabecular volume fraction in radius (2.5 ± 5.1%, p < 0.05) and tibia (2.2 ± 2.2%, p < 0.01). FE estimated bone strength was preserved, but not increased, with PTH 1-34 and zoledronic acid at both sites, whereas it decreased with PTH 1-84 in radius (-2.8 ± 5.8%, p < 0.05) and tibia (-3.9 ± 4.8%, p < 0.001). Conclusively, divergent treatment-specific effects in cortical

  2. 24% Indigenously Prepared Ethylene Diamine Tetra Acetic Acid Compared to Self-Etching Adhesives and their Effect on Shear Bond Strength of Composites in Primary Teeth: An In-vitro Study

    PubMed Central

    Nagar, Priya; Tandil, Yogesh L.; T.P., Chandru; Gupta, Anamika; Kalaria, Devendra; Kumar, Prafful

    2015-01-01

    Background: Over the years, it has been known that 34% phosphoric acid is the benchmark in etchants with the best shear bond strength shown with composites in primary teeth. However, with latest technological advancements and innovations, in order to reduce the number of steps and less damage to the tooth structure, non-rinse conditioner (NRC) & Single-Etch and various other etchants have been tried and tested. These etchants have been found to have shear bond strength comparable to phosphoric acid. In this study, indigenously prepared 24% ethylenediaminetetraacetic acid (EDTA) has been compared with established etchants, as to prove if their shear bond strength was closely related. As it is a well-known fact that EDTA could be less damaging to the enamel during etching and hence can be an alternative for etching of primary teeth. Materials and Methods: For the study 60 caries-free primary molars were used, they were sectioned in the middle, after making area for bonding; the marked area was then etched using different etchants for 30 s. Each of the teeth was then rinsed and bonded with composite resin and thermocycling was done. Shear bond strength testing was done on the composite using Universal Testing Machine. Results: Results of the study showed that phosphoric acid showed the highest bond strength, closely followed by Single Etch (Adper Prompt) and NRC, then by EDTA. Conclusions: About 24% EDTA can be another comparable replacement for phosphoric acid if used with a Single Etch Primer, like Prime and Bond NT on primary teeth. 34% phosphoric acid has the highest bond strength values with composite resin. Single etch followed by NRC has the second and third highest bond strength values, which are comparable to phosphoric acid. PMID:26464540

  3. Atmospheric oxalic acid and related secondary organic aerosols in Qinghai Lake, a continental background site in Tibet Plateau

    NASA Astrophysics Data System (ADS)

    Meng, Jingjing; Wang, Gehui; Li, Jianjun; Cheng, Chunlei; Cao, Junji

    2013-11-01

    Summertime PM2.5 aerosols collected from Qinghai Lake (3200 m a.s.l.), a remote continental site in the northeastern part of Tibetan Plateau, were analyzed for dicarboxylic acids (C2-C11), ketocarboxylic acids and α-dicarbonyals. Oxalic acid (C2) is the dominant dicarboxylic acid in the samples, followed by malonic, succinic and azelaic acids. Total dicarboxylic acids (231 ± 119 ng m-3), ketocarboxylic acids (8.4 ± 4.3 ng m-3), and α-dicarbonyls (2.7 ± 2.1 ng m-3) at the Tibetan background site are 2-5 times less than those detected in lowland areas such as 14 Chinese megacities. Compared to those in other urban and marine areas enhancements in relative abundances of C2/total diacids and diacids-C/WSOC of the PM2.5 samples suggest that organic aerosols in the region are more oxidized due to strong solar radiation. Molecular compositions and air mass trajectories demonstrate that the above secondary organic aerosols in the Qinghai Lake atmosphere are largely derived from long-range transport. Ratios of oxalic acid, glyoxal and methylglyoxal to levoglucosan in PM2.5 aerosols emitted from household burning of yak dung, a major energy source for Tibetan in the region, are 30-400 times lower than those in the ambient air, which further indicates that primary emission from biomass burning is a negligible source of atmospheric oxalic acid and α-dicarbonyls at this background site.

  4. Isotope geochemistry of waters affected by acid mine drainage in old labour sites (SE, Spain).

    NASA Astrophysics Data System (ADS)

    Pérez-Sirvent, Carmen; Martinez-Sanchez, Maria Jose; Garcia-Lorenzo, Maria Luz; Agudo, Ines; Hernandez-Cordoba, Manuel; Recio, Clemente

    2015-04-01

    The ore deposits of this zone have iron, lead and zinc as the main metal components. Iron is present in oxides, hydroxides, sulfides, sulfates, carbonates, and silicates; lead and zinc occur in sulfides (galena and sphalerite, respectively), carbonates, sulfates, and lead or zinc-bearing (manganese, iron) oxides. Mining started with the Romans and activity peaked in the second half of the 19th century and throughout the 20th century until the 1980's. From 1940 to 1957, mineral concentration was made by froth flotation and, prior to this, by gravimetric techniques. The mining wastes, or tailings, with a very fine particle size were deposited inland (tailings dams) and, since 1957, huge releases were made in directly the sea coast. The objective of this work was to evaluate processes affecting waters from abandoned mine sites by way of stable isotopic analysis, particularly H and O stable isotopes from water and S and O from dissolved sulfates. Several common chemical and physical processes, such as evaporation, water-rock interaction and mixing could alter water isotopic composition. Evaporation, which causes an enrichment in δD and δ18O in the residual water, is an important process in semiarid areas. The results obtained indicate that, for sites near the coast, waters are meteoric, and marine infiltration only takes place in the deepest layers near the shore or if water remains stagnated in sediments with low permeability. The main source of sulfate was the oxidation of sulfides, resulting in the liberation of acid, sulfate and metals. In order to assess the mechanism responsible for sulfide oxidation, the stoichiometric isotope balance model and the general isotope balance model were tested, suggesting that the oxidation via Fe3+ was predominant in the surface, and controlled by A. ferrooxidans, while at depth, sulfate reduction occurred.

  5. Early smelter sites: A neglected chapter in the history and geography of acid rain in the United States

    NASA Astrophysics Data System (ADS)

    Quinn, M.-L.

    Dominant spatial and temporal theories of acid rain in the U.S. are identified, followed by brief comments on how historical data have generally been used in modern acid rain research. A frequently-cited 1982 article by E.B. Cowling is examined, one that has influenced much thinking on the history of acid rain. The article overlooks early American smelters, however, and the role they played in the true history and geography of acid rain in the United States. Continuing with this theme, a connection is established between acid rain and turn-of-the-century smelter smoke problems. Literature on the latter subject is discussed, and American and German examples are given. A beginning is then made on writing acid rain's neglected chapter, focusing on Tennessee's Copper Basin (Ducktown District) where copper smelting dates back to the 1850s. A short historical overview of this area's smelting operations is given, with particular attention to the air pollution and other environmental problems resulting from large emissions of sulfur dioxide. Five additional early smelter sites for potential study are mentioned as well. The paper concludes with some observations regarding the way in which expanded research of early smelter sites could affect the general perception of acid rain in the U.S. It is also suggested that such research might contribute to a better atmosphere for making decisions and policies pertaining to the phenomenon as it exists today.

  6. CHARACTERIZATION OF INDIVIDUAL CHEMICAL REACTIONS CONSUMING ACID DURING NUCLEAR WASTE PROCESSING AT THE SAVANNAH RIVER SITE - 136B

    SciTech Connect

    Koopman, D.; Pickenheim, B.; Lambert, D.; Newell, J.; Stone, M.

    2009-09-02

    Conversion of legacy radioactive high-level waste at the Savannah River Site into a stable glass waste form involves a chemical pretreatment process to prepare the waste for vitrification. Waste slurry is treated with nitric and formic acids to achieve certain goals. The total quantity of acid added to a batch of waste slurry is constrained by the catalytic activity of trace noble metal fission products in the waste that can convert formic acid into hydrogen gas at many hundreds of times the radiolytic hydrogen generation rate. A large block of experimental process simulations were performed to characterize the chemical reactions that consume acid prior to hydrogen generation. The analysis led to a new equation for predicting the quantity of acid required to process a given volume of waste slurry.

  7. Influence of methoxy-substituents on the strength of Br … Br type II halogen bonds in bromobenzoic acid

    NASA Astrophysics Data System (ADS)

    Raffo, Pablo A.; Marcolongo, Juan P.; Funes, Alejandro V.; Slep, Leonardo D.; Baggio, Ricardo F.; Cukiernik, Fabio D.

    2016-03-01

    4-bromo-3,5-di(methoxy)benzoic acid (I) crystallizes in the monoclinic C2/c space group, a = 22.3405 (6) Å, b = 4.85142 (14) Å, c = 18.1583 (5) Å, β = 93.086 (2)°. The crystal structure shows head-to-head dimeric units linked via type II Br … Br interactions as well as Br … π and weak H-bonding interactions. The whole structure exhibits features similar to those of the parent 4-bromobenzoic acid (II), most notably the overall geometrical features involved in the Br … Br type II interactions. Both structures display comparable C-Br … Br angles (θ1 = 98.3 and 91.6° and θ2 = 163.0 and 163.5° for (I) and (II) respectively), but the Br … Br distance is significantly shorter in (I) (3.58 Å) than in (II) (3.81 Å). QM computations provide the magnitude of the intermolecular interactions present in both (I) and (II), and allow disclosing the individual covalent and electrostatic contributions to the Br⋯Br halogen bond in terms of interaction energies, electrostatic potentials, and a molecular orbital (MO) analysis.

  8. Iodine 125-lysergic acid diethylamide binds to a novel serotonergic site on rat choroid plexus epithelial cells

    SciTech Connect

    Yagaloff, K.A.; Hartig, P.R.

    1985-12-01

    /sup 125/I-Lysergic acid diethylamide (/sup 125/I-LSD) binds with high affinity to serotonergic sites on rat choroid plexus. These sites were localized to choroid plexus epithelial cells by use of a novel high resolution stripping film technique for light microscopic autoradiography. In membrane preparations from rat choroid plexus, the serotonergic site density was 3100 fmol/mg of protein, which is 10-fold higher than the density of any other serotonergic site in brain homogenates. The choroid plexus site exhibits a novel pharmacology that does not match the properties of 5-hydroxytryptamine-1a (5-HT1a), 5-HT1b, or 5-HT2 serotonergic sites. /sup 125/I-LSD binding to the choroid plexus site is potently inhibited by mianserin, serotonin, and (+)-LSD. Other serotonergic, dopaminergic, and adrenergic agonists and antagonists exhibit moderate to weak affinities for this site. The rat choroid plexus /sup 125/I-LSD binding site appears to represent a new type of serotonergic site which is located on non-neuronal cells in this tissue.

  9. The two active sites in human branched-chain alpha-keto acid dehydrogenase operate independently without an obligatory alternating-site mechanism.

    PubMed

    Li, Jun; Machius, Mischa; Chuang, Jacinta L; Wynn, R Max; Chuang, David T

    2007-04-20

    A long standing controversy is whether an alternating activesite mechanism occurs during catalysis in thiamine diphosphate (ThDP)-dependent enzymes. We address this question by investigating the ThDP-dependent decarboxylase/dehydrogenase (E1b) component of the mitochondrial branched-chain alpha-keto acid dehydrogenase complex (BCKDC). Our crystal structure reveals that conformations of the two active sites in the human E1b heterotetramer harboring the reaction intermediate are identical. Acidic residues in the core of the E1b heterotetramer, which align with the proton-wire residues proposed to participate in active-site communication in the related pyruvate dehydrogenase from Bacillus stearothermophilus, are mutated. Enzyme kinetic data show that, except in a few cases because of protein misfolding, these alterations are largely without effect on overall activity of BCKDC, ruling out the requirement of a proton-relay mechanism in E1b. BCKDC overall activity is nullified at 50% phosphorylation of E1b, but it is restored to nearly half of the pre-phosphorylation level after dissociation and reconstitution of BCKDC with the same phosphorylated E1b. The results suggest that the abolition of overall activity likely results from the specific geometry of the half-phosphorylated E1b in the BCKDC assembly and not due to a disruption of the alternating active-site mechanism. Finally, we show that a mutant E1b containing only one functional active site exhibits half of the wild-type BCKDC activity, which directly argues against the obligatory communication between active sites. The above results provide evidence that the two active sites in the E1b heterotetramer operate independently during the ThDP-dependent decarboxylation reaction. PMID:17329260

  10. Effects of background electrolytes and ionic strength on enrichment of Cd(II) ions with magnetic graphene oxide-supported sulfanilic acid.

    PubMed

    Hu, Xin-jiang; Liu, Yun-guo; Zeng, Guang-ming; You, Shao-hong; Wang, Hui; Hu, Xi; Guo, Yi-ming; Tan, Xiao-fei; Guo, Fang-ying

    2014-12-01

    To elucidate the influence mechanisms of background electrolytes and ionic strength on Cd(II) removal, the adsorption of Cd(II) onto magnetic graphene oxide-supported sulfanilic acid (MGO-SA) in aqueous solutions containing different types and concentrations of background electrolytes was studied. The results indicate that Cd(II) adsorption was strongly dependent on pH and could be strongly affected by background electrolytes and ionic strength. The Cd(II) removal was decreased with the presence of background electrolyte cations (Na(+), K(+), Ca(2+), Mg(2+), Mn(2+), Zn(2+), and Ni(2+)), and the divalent cations exerted more obvious influences on the Cd(II) uptake than the monovalent cations at pH 6. Both Cl(-) and NO3(-) had negative effects on Cd(II) adsorption because they can form water-soluble metal-anion complexes with Cd(II) ions. The presence of 0.01molL(-1) Na3PO4 reduced the removal percentage of Cd(II) at pH<5 but extremely enhanced the Cd(II) removal when the pH>5. The Cd(II) adsorption was sensitive to changes in the concentration of NaCl, NaNO3, NaClO4, and Na3PO4. Besides, the adsorption isotherm of Cd(II) onto MGO-SA could be well described by the Freundlich model and was also influenced by the type of background electrolyte ions and the ionic strength.

  11. Identification of gamma-aminobutyric acid and its binding sites in Caenorhabditis elegans

    SciTech Connect

    Schaeffer, J.M.; Bergstrom, A.R.

    1988-01-01

    Gamma-aminobutyric acid (GABA), glutamate decarboxylase and GABA-transaminase were identified in the nematode Caenorhabditis elegans. The concentration of GABA in C. elegans is approximately 10-fold lower than the concentration of GABA in rat brain. Glutamate decarboxylase and GABA-transaminase, the GABA anabolic and catabolic enzymes, are also present in C. elegans. Crude membrane fractions were prepared from C. elegans and used to study specific (/sup 3/H) GABA binding sites. GABA binds to C. elegans membranes with high affinity and low capacity. Muscimol is a competitive inhibitor of specific GABA binding with a K/sub I/ value of 120 nM. None of the other GABA agonists or antagonists inhibited greater than 40% of the specific GABA binding at concentrations up to 10/sup -4/M. Thirteen spider venoms were examined as possible GABA agonists or antagonists, the venom from Calilena agelenidae inhibits specific GABA binding with a K/sub I/ value of 6 nl/ml. These results suggest that GABA has a physiological role as a neurotransmitter in C. elegans.

  12. Interaction of pseudolaric acid B with the colchicine site of tubulin.

    PubMed

    Sarkar, Taradas; Nguyen, Tam Luong; Su, Zhi-Wei; Hao, Jun; Bai, Ruoli; Gussio, Rick; Qiu, Samuel X; Hamel, Ernest

    2012-08-15

    We purified pseudolaric acid B (PAB) from the root and stem bark of Pseudolarix kaempferi (Lindl.) Gorden. Confirming previous findings, we found that the compound had high nanomolar IC₅₀ antiproliferative effects in several cultured cell lines, causing mitotic arrest and the disappearance of intracellular microtubules. PAB strongly inhibited tubulin assembly (IC₅₀, 1.1 μM) but weakly inhibited the binding of colchicine to tubulin, as demonstrated by fluorescence and with [³H]colchicine. Kinetic analysis demonstrated that the mechanism of inhibition was competitive, with an apparent K(i) of 12-15 μM. Indirect studies demonstrated that PAB bound rapidly to tubulin and dissociated more rapidly from tubulin than the colchicine analog 2-methoxy-5-(2',3',4'-trimethoxyphenyl)tropone, whose complex with tubulin is known to have a half-life of 17s at 37 °C. We modeled PAB into the colchicine site of tubulin, using the crystal structure 1SA0 that contains two αβ-tubulin heterodimers, both bound to a colchicinoid and to a stathmin fragment. The binding model of PAB revealed common pharmacophoric features between PAB and colchicinoids, not readily apparent from their chemical structures. PMID:22634405

  13. High-Temperature Decomposition of Brønsted Acid Sites in Gallium-Substituted Zeolites

    SciTech Connect

    K Al-majnouni; N Hould; W Lonergan; D Vlachos; R Lobo

    2011-12-31

    The dehydroxylation of Broensted acid sites (BAS) in Ga-substituted zeolites was investigated at temperatures up to 850 C using X-ray absorption spectroscopy (XAS), Fourier transform infrared spectroscopy (FTIR), and mass spectrometry-temperature programmed desorption (MS-TPD). X-ray absorption near-edge spectroscopy (XANES) revealed that the majority of gallium has tetrahedral coordination even after complete dehydroxylation. The interatomic gallium-oxygen distance and gallium coordination number determined by extended X-ray absorption fine structure (EXAFS) are consistent with gallium in tetrahedral coordination at low T (< 550 C). Upon heating Ga-Beta and Ga-ZSM5 to 850 C, analysis of the EXAFS showed that 70 and 80% of the gallium was still in tetrahedral coordination. The remainder of the gallium was found to be in octahedral coordination. No trigonal Ga atoms were observed. FTIR measurements carried out at similar temperatures show that the intensity of the OH vibration due to BAS has been eliminated. MS-TPD revealed that hydrogen in addition to water evolved from the samples during dehydroxylation. This shows that dehydrogenation in addition to dehydration is a mechanism that contributes to BAS decomposition. Dehydrogenation was further confirmed by exposing the sample to hydrogen to regenerate some of the BAS as monitored by FTIR and MS-TPD.

  14. Compromised Osseous Healing of Dental Extraction Sites in Zoledronic Acid-Treated Dogs

    PubMed Central

    Allen, Matthew R.; Kubek, Daniel J.; Burr, David B.; Ruggiero, Salvatore L.; Chu, Tien-Min Gabriel

    2010-01-01

    PURPOSE The goal of this study was to document how treatment with a bisphosphonate affects the bone tissue following dental extraction. METHODS Skeletally mature female beagle dogs were either untreated controls (CON) or treated with intravenous zoledronic acid (ZOL). Following the extraction of the 4th premolars, healing was allowed for 4 or 8 weeks. Properties of the extraction site were assessed using micro-computed tomography (micro-CT) and dynamic histomorphometry. RESULTS The initial infilling of the extraction socket with bone was not affected by ZOL but subsequent removal of this bone was significantly suppressed compared to CON. After 8-weeks of healing, the alveolar cortical bone adjacent to the extraction socket had a remodeling rate of ~50%/year in CON animals while ZOL-treated animals had a rate of < 1%/year. One ZOL-treated animal developed exposed bone post-extraction which eventually led to the formation of a sequestrum. Assessment of the sequestrum with micro-CT and histology showed that it had features consistent with those reported in humans with osteonecrosis of the jaw. CONCLUSIONS These results, showing significantly compromised post-extraction osseous healing as well as presence of exposed bone and development of a sequestrum in one ZOL animal, provide a building block toward understanding the pathophysiology of osteonecrosis of the jaw. PMID:20458574

  15. 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.

  16. An Aqueous Thermodynamic Model for the Complexation of Sodium and Strontium with Organic Chelates valid to High Ionic Strength. I. Ethylenedinitrilotetraacetic acid (EDTA)

    SciTech Connect

    Felmy, Andrew R.; Mason, Marvin J.

    2003-04-01

    An aqueous thermodynamic model is developed, which accurately describes the effects of Na+ complexation, ionic strength, carbonate concentration, and temperature on the complexation of Sr2+ by ethylenedinitrilotetraacetic acid (EDTA) under basic conditions. The model is developed from the analysis of literature data on apparent equilibrium constants, enthalpies, and heat capacities; as well as on an extensive set of solubility data on SrCO3(c) in the presence of EDTA obtained as part of this study. The solubility data for SrCO3(c) were obtained in solutions ranging in Na2CO3 concentration from 0.01m to 1.8m, in NaNO3 concentration from 0 to 5m, and at temperatures extending to 75?C. The final aqueous thermodynamic model is based upon the equations of Pitzer and requires the inclusion of a NaEDTA3- species. An accurate model for the ionic strength dependence of the ion-interaction coefficients for the SrEDTA2- and NaEDTA3- aqueous species allows the extrapolation of standard state equilibrium constants for these species which are significantly different from the 0.1m reference state values available in the literature. The final model is tested by application to chemical systems containing competing metal ions (i.e., Ca2+) to further verify the proposed model and indicate the applicability of the model parameters to chemical systems containing other divalent metal-EDTA complexes.

  17. ESTIMATES OF CLOUD WATER DEPOSITION AT MOUNTAIN DEPOSITION AT MOUNTAIN ACID DEPOSITION PROGRAM SITES IN THE APPALACHIAN MOUNTAINS

    EPA Science Inventory

    Cloud water deposition was estimated at three high elevation sites in the Appalachian Mountains of the eastern United States (Whiteface Mountain, NY, Whitetop Mountain, VA, and Clingrnan's Dome, TN) from 1994 through 1999 as part of the Mountain Acid Deposition Program (MADPro). ...

  18. Non-thermal plasma-assisted NOx reduction over Na-Y zeolites: The promotional effect of acid sites

    SciTech Connect

    Kwak, Ja Hun; Peden, Charles HF; Szanyi, Janos

    2006-06-01

    The effect of acid sites on the catalytic activities of a series of H+-modified Na-Y zeolites was investigated in the non-thermal plasma assisted NOx reduction reaction using a simulated diesel engine exhaust gas mixture. The acid sites were formed by NH4+ ion exchange and subsequent heat treatment of a NaY zeolite. The catalytic activities of these H+-modified NaY zeolites significantly increased with the number of acid sites. This NOx conversion increase was correlated with the decrease in the amount of unreacted NO2. The increase in the number of acid sites did not change the NO level, it stayed constant. Temperature programmed desorption following NO2 adsorption showed the appearance of a high temperature desorption peak at 453 K in addition to the main desorption feature of 343 K observed for the base Na-Y. The results of both the IR and TPD experiments revealed the formation of crotonaldehyde, resulting from condensation reaction of adsorbed acetaldehyde. Strong adsorptions of both NOx and hydrocarbon species are proposed to be responsible for the higher catalytic activity of H+-modified Na-Y zeolites in comparison to the base NaY material

  19. Shear bond strength and debonding characteristics of metal and ceramic brackets bonded with conventional acid-etch and self-etch primer systems: An in-vivo study

    PubMed Central

    Mirzakouchaki, Behnam; Sharghi, Reza; Shirazi, Samaneh; Moghimi, Mahsan; Shahrbaf, Shirin

    2016-01-01

    Background Different in-vitro studies have reported various results regarding shear bond strength (SBS) of orthodontic brackets when SEP technique is compared to conventional system. This in-vivo study was designed to compare the effect of conventional acid-etching and self-etching primer adhesive (SEP) systems on SBS and debonding characteristics of metal and ceramic orthodontic brackets. Material and Methods 120 intact first maxillary and mandibular premolars of 30 orthodontic patients were selected and bonded with metal and ceramic brackets using conventional acid-etch or self-etch primer system. The bonded brackets were incorporated into the wire during the study period to simulate the real orthodontic treatment condition. The teeth were extracted and debonded after 30 days. The SBS, debonding characteristics and adhesive remnant indices (ARI) were determined in all groups. Results The mean SBS of metal brackets was 10.63±1.42 MPa in conventional and 9.38±1.53 MPa in SEP system, (P=0.004). No statistically significant difference was noted between conventional and SEP systems in ceramic brackets. The frequency of 1, 2 and 3 ARI scores and debonding within the adhesive were the most common among all groups. No statistically significant difference was observed regarding ARI or failure mode of debonded specimens in different brackets or bonding systems. Conclusions The SBS of metal brackets bonded using conventional system was significantly higher than SEP system, although the SBS of SEP system was clinically acceptable. No significant difference was found between conventional and SEP systems used with ceramic brackets. Total SBS of metal brackets was significantly higher than ceramic brackets. Due to adequate SBS of SEP system in bonding the metal brackets, it can be used as an alternative for conventional system. Key words:Shear bond strength, Orthodontic brackets, Adhesive remnant index, self-etch. PMID:26855704

  20. pH and ionic strength effects on the binding constant between a nitrogen-containing polycyclic aromatic compound and humic acid.

    PubMed

    Chang, Kuei-Chen; Lee, Chon-Lin; Hsieh, Ping-Chieh; Brimblecombe, Peter; Kao, Shu-Min

    2015-09-01

    Polycyclic aromatic compounds (PACs) are widespread environmental pollutants with a high potential to act as human carcinogens and mutagens. The behavior of PACs is significantly affected by their interactions with dissolved organic matter (DOM), such as their transport, solubility, bioavailability, and bioaccumulation in the aquatic environment. Being a basic PAC, benzo(h)quinoline (BQ) is the dominant species, as the solution's pH value is higher than BQ's pK a (pK a of BQ = 4.2). In contrast, benzo(h)quinolinium (BQH(+)) is the major species, as the solution's pH value is lower than its pK a. The binding constant (K DOC), measured by fluorescence quenching, between BQ/BQH(+) and Leonardite humic acid (LHA) would decrease 70 to 95 % and 20 to 90 % when increasing the ionic strength in acidic and neutral to basic conditions, respectively. The results can be attributed to the added cation (Na(+) and Mg(2+)), which forms a bridge with LHA and enhances the intramolecular reaction among these functional groups, therefore inducing the coiling up within the LHA molecule. In addition, the decrease of the K DOC with added MgCl2/MgSO4 (75-95 %) is higher than that with added NaCl/Na2SO4 (20-75 %), indicating that the K DOC was affected by the charge density of cations. The fluorescence intensity of BQH(+) in the absence of LHA (F 0) was found to decay only in the acidic solution with Cl(-), suggesting that Cl(-) might be a heavy atom serving as a quencher in an acidic solution. PMID:25940463

  1. Uptake of gamma-aminobutyric acid and L-glutamic acid by synaptosomes from postmortem human cerebral cortex: multiple sites, sodium dependence and effect of tissue preparation.

    PubMed

    Dodd, P R; Watson, W E; Morrison, M M; Johnston, G A; Bird, E D; Cowburn, R F; Hardy, J A

    1989-06-26

    The uptake of gamma-aminobutyric acid (GABA) and L-glutamic acid by synaptosomes prepared from frozen postmortem human brain was shown to be effected via distinct high and low affinity sites. At approximately 17 h postmortem delay, the kinetic parameters for GABA uptake were: high affinity site, Km 7.1 +/- 2.5 microM, Vmax 18.7 +/- 4.8 nmol.min-1 per 100 mg protein; low affinity site, Km 2 +/- 1 mM, Vmax 425 +/- 250 nmol.min-1 per 100 mg protein (means +/- S.E.M., n = 13). Kinetic parameters for L-glutamate uptake were: high affinity site, Km 7.5 +/- 1.0 microM, Vmax 85 +/- 8 nmol.min-1 per 100 mg protein; low affinity site, Km 1.8 +/- 1.2 mM. Vmax 780 +/- 175 nmol.min-1 per 100 mg protein (n = 11). A detailed kinetic analysis of high affinity GABA uptake was performed over a range of sodium ion concentrations. The results were consistent with a coupling ratio of one Na+ ion to one GABA molecule; a similar result was found with rat brain synaptosomes. However, rat and human synaptosomes differed in the degree to which the substrate affinity of the high affinity GABA uptake site varied with decreasing Na+ ion concentration. High affinity GABA uptake was markedly affected by the method used to freeze and divide the tissue, but did not vary greatly in different cortical regions. There was some decline of high affinity GABA uptake activity with postmortem delay, apparently due to a loss of sites rather than a change in site affinity.

  2. Forced Ambiguity of the Leucine Codons for Multiple-Site-Specific Incorporation of a Noncanonical Amino Acid

    PubMed Central

    Kwon, Inchan; Choi, Eun Sil

    2016-01-01

    Multiple-site-specific incorporation of a noncanonical amino acid into a recombinant protein would be a very useful technique to generate multiple chemical handles for bioconjugation and multivalent binding sites for the enhanced interaction. Previously combination of a mutant yeast phenylalanyl-tRNA synthetase variant and the yeast phenylalanyl-tRNA containing the AAA anticodon was used to incorporate a noncanonical amino acid into multiple UUU phenylalanine (Phe) codons in a site-specific manner. However, due to the less selective codon recognition of the AAA anticodon, there was significant misincorporation of a noncanonical amino acid into unwanted UUC Phe codons. To enhance codon selectivity, we explored degenerate leucine (Leu) codons instead of Phe degenerate codons. Combined use of the mutant yeast phenylalanyl-tRNA containing the CAA anticodon and the yPheRS_naph variant allowed incorporation of a phenylalanine analog, 2-naphthylalanine, into murine dihydrofolate reductase in response to multiple UUG Leu codons, but not to other Leu codon sites. Despite the moderate UUG codon occupancy by 2-naphthylalaine, these results successfully demonstrated that the concept of forced ambiguity of the genetic code can be achieved for the Leu codons, available for multiple-site-specific incorporation. PMID:27028506

  3. Dynamic HypA zinc site is essential for acid viability and proper urease maturation in Helicobacter pylori

    PubMed Central

    Johnson, Ryan C.; Hu, Heidi Q.; Merrell, D. Scott; Maroney, Michael J.

    2015-01-01

    Helicobacter pylori requires urease activity in order to survive in the acid environment of the human stomach. Urease is regulated in part by nickelation, a process that requires the HypA protein, which is a putative nickel metallochaperone that is generally associated with hydrogenase maturation. However, in H. pylori, HypA plays a dual role. In addition to an N-terminal nickel binding site, HypA proteins also contain a structural zinc site that is coordinated by two rigorously conserved CXXC sequences, which in H. pylori are flanked by His residues. These structural Zn sites are known to be dynamic, converting from Zn(Cys)4 centers at pH 7.2 to Zn(Cys)2(His)2 centers at pH 6.3 in the presence of Ni(II) ions. In this study, mutant strains of H. pylori that express zinc site variants of the HypA protein are used to show that the structural changes in the zinc site are important for the acid viability of the bacterium, and that a reduction in acid viability in these variants can be traced in large measure to deficient urease activity. This in turn leads to a model that connects the Zn(Cys)4 coordination to urease maturation. PMID:25608738

  4. Effects of sex and site on amino acid metabolism enzyme gene expression and activity in rat white adipose tissue

    PubMed Central

    Arriarán, Sofía; Agnelli, Silvia; Remesar, Xavier; Fernández-López, José Antonio

    2015-01-01

    Background and Objectives. White adipose tissue (WAT) shows marked sex- and diet-dependent differences. However, our metabolic knowledge of WAT, especially on amino acid metabolism, is considerably limited. In the present study, we compared the influence of sex on the amino acid metabolism profile of the four main WAT sites, focused on the paths related to ammonium handling and the urea cycle, as a way to estimate the extent of WAT implication on body amino-nitrogen metabolism. Experimental Design. Adult female and male rats were maintained, undisturbed, under standard conditions for one month. After killing them under isoflurane anesthesia. WAT sites were dissected and weighed. Subcutaneous, perigonadal, retroperitoneal and mesenteric WAT were analyzed for amino acid metabolism gene expression and enzyme activities. Results. There was a considerable stability of the urea cycle activities and expressions, irrespective of sex, and with only limited influence of site. Urea cycle was more resilient to change than other site-specialized metabolic pathways. The control of WAT urea cycle was probably related to the provision of arginine/citrulline, as deduced from the enzyme activity profiles. These data support a generalized role of WAT in overall amino-N handling. In contrast, sex markedly affected WAT ammonium-centered amino acid metabolism in a site-related way, with relatively higher emphasis in males’ subcutaneous WAT. Conclusions. We found that WAT has an active amino acid metabolism. Its gene expressions were lower than those of glucose-lipid interactions, but the differences were quantitatively less important than usually reported. The effects of sex on urea cycle enzymes expression and activity were limited, in contrast with the wider variations observed in other metabolic pathways. The results agree with a centralized control of urea cycle operation affecting the adipose organ as a whole. PMID:26587356

  5. Amino acid sequences of neuropeptides in the sinus gland of the land crab Cardisoma carnifex: a novel neuropeptide proteolysis site.

    PubMed

    Newcomb, R W

    1987-08-01

    The sinus gland is a major neurosecretory structure in Crustacea. Five peptides, labeled C, D, E, F, and I, isolated from the sinus gland of the land crab have been hypothesized to arise from the incomplete proteolysis at two internal sites on a single biosynthetic intermediate peptide "H", based on amino acid composition additivities and pulse-chase radiolabeling studies. The presence of only a single major precursor for the sinus gland peptides implies that peptide H may be synthesized on a common precursor with crustacean hyperglycemic hormone forms, "J" and "L," and a peptide, "K," similar to peptides with molt inhibiting activity. Here I report amino acid sequences of these peptides. The amino terminal sequence of the parent peptide, H, (and the homologous fragments) proved refractory to Edman degradation. Data from amino acid analysis and carboxypeptidase digestion of the naturally occurring fragments and of fragments produced by endopeptidase digestion were used together with Edman degradation to obtain the sequences. Amino acid analysis of fragments of the naturally occurring "overlap" peptides (those produced by internal cleavage at one site on H) was used to obtain the sequences across the cleavage sites. The amino acid sequence of the land crab peptide H is Arg-Ser-Ala-Asp-Gly-Phe-Gly-Arg-Met-Glu-Ser-Leu-Leu-Thr-Ser-Leu-Arg-Gly- Ser-Ala-Glu- Ser-Pro-Ala-Ala-Leu-Gly-Glu-Ala-Ser-Ala-Ala-His-Pro-Leu-Glu. In vivo cleavage at one site involves excision of arginine from the sequence Leu-Arg-Gly, whereas cleavage at the other site involves excision of serine from the sequence Glu-Ser-Leu. Proteolysis at the latter sequence has not been previously reported in intact secretory granules. The aspartate at position 4 is possibly covalently modified.

  6. Retinoic acid signalling centres in the avian embryo identified by sites of expression of synthesising and catabolising enzymes.

    PubMed

    Blentic, Aida; Gale, Emily; Maden, Malcolm

    2003-05-01

    Retinoic acid is an important signalling molecule in the developing embryo, but its precise distribution throughout development is very difficult to determine by available techniques. Examining the distribution of the enzymes by which it is synthesised by using in situ hybridisation is an alternative strategy. Here, we describe the distribution of three retinoic acid synthesising enzymes and one retinoic acid catabolic enzyme during the early stages of chick embryogenesis with the intention of identifying localized retinoic acid signalling regions. The enzymes involved are Raldh1, Raldh2, Raldh3, and Cyp26A1. Although some of these distributions have been described before, here we assemble them all in one species and several novel sites of enzyme expression are identified, including Hensen's node, the cardiac endoderm, the presumptive pancreatic endoderm, and the dorsal lens. This study emphasizes the dynamic pattern of expression of the enzymes that control the availability of retinoic acid as well as the role that retinoic acid plays in the development of many regions of the embryo throughout embryogenesis. This strategy provides a basis for understanding the phenotypes of retinoic acid teratology and retinoic acid-deficiency syndromes.

  7. Participation and strength of interaction of lysine 95(beta) in the polymerization of hemoglobin S as determined by its site-directed substitution by isoleucine.

    PubMed

    Himanen, J P; Schneider, K; Chait, B; Manning, J M

    1995-06-01

    The role of Lys-95(beta), which is on the exterior of the hemoglobin (HbS) tetramer, in the aggregation process has been addressed because there is a lack of agreement on its importance. The early studies on the aggregation of HbS in the presence of other mutant hemoglobins are consistent with the subsequent electron microscopic studies in demonstrating the participation of Lys-95(beta) in gelation; the results of the crystal structure do not agree with these conclusions. Therefore, with the objective of clarifying its role we have carried out site-directed substitution of Lys-95(beta) to an isoleucine residue. The mutation was introduced by polymerase chain reaction recombination methodology, and the absence of other mutations in the beta-globin gene was established by sequencing the gene in its entirety. The recombinant mutant hemoglobin was expressed in yeast and characterized by peptide mapping and sequencing, which demonstrated that the only different tryptic peptide had the Ile substitution at position 95(beta). The recombinant hemoglobin had the correct amino acid composition and molecular weight by mass spectrometric analysis. It was also pure as judged by isoelectric focusing. It was fully functional because it had an average Hill coefficient of 3.1 and responded normally to the allosteric regulators, chloride, 2,3-diphosphoglycerate, and inositol hexaphosphate. Of particular interest was the finding that this hemoglobin mutant aggregated at a concentration of about 40 g/dl, nearly twice that at which HbS itself aggregated (24 g/dl). Therefore, Lys-95(beta) has a very important role in the aggregation process and is a good candidate site for the design of a therapeutic agent for sickle cell anemia. PMID:7775447

  8. Predicting Effects of Corrosion Erosion of High Strength Steel Pipelines Elbow on CO2-Acetic Acid (HAc) Solution

    NASA Astrophysics Data System (ADS)

    Asmara, Y. P.; Ismail, M. F.; Giok Chui, L.; Halimi, Jamiludin

    2016-02-01

    Simultaneously effect of erosion combined with corrosion becomes the most concern in oil and gas industries. It is due to the fast deterioration of metal as effects of solid particles mixed with corrosive environment. There are many corrosion software to investigate possible degradation mechanisms developed by researchers. They are using many combination factors of chemical reactions and physical process. However effects of CO2 and acid on pipelines orientations are still remain uncovered in their simulation. This research will investigate combination effects of CO2 and HAc on corrosion and erosion artificial environmental containing sands particles in 45°, 90° and 180° elbow pipelines. The research used theoretical calculations combined with experiments for verification. The main concerns are to investigate the maximum erosion corrosion rate and maximum shear stress at the surface. Methodology used to calculate corrosion rate are Linear Polarization Resistance (LPR) and weight loss. The results showed that at 45°, erosion rate is the more significant effects in contributing degradation of the metal. The effects of CO2 and HAc gave significant effects when flow rate of the solution are high which reflect synergism effects of solid particles and those chemical compositions.

  9. Acid-base strength and acidochromism of some dimethylamino-azinium iodides. An integrated experimental and theoretical study.

    PubMed

    Benassi, Enrico; Carlotti, Benedetta; Fortuna, Cosimo G; Barone, Vincenzo; Elisei, Fausto; Spalletti, Anna

    2015-01-15

    The effects of pH on the spectral properties of stilbazolium salts bearing dimethylamino substituents, namely, trans isomers of the iodides of the dipolar E-[2-(4-dimethylamino)styryl]-1-methylpyridinium, its branched quadrupolar analogue E,E-[2,6-di-(p-dimethylamino)styryl]-1-methylpyridinium, and three analogues, chosen to investigate the effects of the stronger quinolinium acceptor, the longer butadiene π bridge, or both, were investigated through a joint experimental and computational approach. A noticeable acidochromism of the absorption spectra (interesting for applications) was observed, with the basic and protonated species giving intensely colored and transparent solutions, respectively. The acid–base equilibrium constants for the protonation of the dimethylamino group in the ground state (pKa) were experimentally derived. Theoretical calculations according to the thermodynamic Born-Haber cycle provided pKa values in good agreement with the experimental values. The very low fluorescence yield did not allow a direct investigation of the changes in the acid-base properties in the excited state (pKa*) by fluorimetric titrations. Their values were derived by quantum-mechanical calculations and estimated experimentally on the basis of the Förster cycle.

  10. Gamma-aminobutyric acid-modulated benzodiazepine binding sites in bacteria

    SciTech Connect

    Lummis, S.C.R.; Johnston, G.A.R. ); Nicoletti, G. ); Holan, G. )

    1991-01-01

    Benzodiazepine binding sites, which were once considered to exist only in higher vertebrates, are here demonstrated in the bacteria E. coli. The bacterial ({sup 3}H)diazepam binding sites are modulated by GABA; the modulation is dose dependent and is reduced at high concentrations. The most potent competitors of E.Coli ({sup 3}H)diazepam binding are those that are active in displacing ({sup 3}H)benzodiazepines from vertebrate peripheral benzodiazepine binding sites. These vertebrate sites are not modulated by GABA, in contrast to vertebrate neuronal benzodiazepine binding sites. The E.coli benzodiazepine binding sites therefore differ from both classes of vertebrate benzodiazepine binding sites; however the ligand spectrum and GABA-modulatory properties of the E.coli sites are similar to those found in insects. This intermediate type of receptor in lower species suggests a precursor for at least one class of vertebrate benzodiazepine binding sites may have existed.

  11. Complex coacervation of hyaluronic acid and chitosan: effects of pH, ionic strength, charge density, chain length and the charge ratio.

    PubMed

    Kayitmazer, A B; Koksal, A F; Kilic Iyilik, E

    2015-11-28

    Hyaluronic acid (HA) and chitosan (CH) can form nanoparticles, hydrogels, microspheres, sponges, and films, all with a wide range of biomedical applications. This variety of phases reflects the multiple pathways available to HA/CH complexes. Here, we use turbidimetry, dynamic light scattering, light microscopy and zeta potential measurements to show that the state of the dense phase depends on the molar ratio of HA carboxyl to CH amines, and is strongly dependent on their respective degrees of ionization, α and β. Due to the strong charge complementarity between HA and CH, electrostatic self-assembly takes place at very acidic pH, but is almost unobservable at ionic strength (I) ≥ 1.5 M NaCl. All systems display discontinuity in the I-dependence of the turbidity, corresponding to a transition from coacervates to flocculates. An increase in either polymer chain length or charge density enhances phase separation. Remarkably, non-stoichiometric coacervate suspensions form at zeta potentials far away from zero. This result is attributed to the entropic effects of chain semi-flexibility as well as to the charge mismatch between the two biopolymers. PMID:26406548

  12. Observation of multiple, identical binding sites in the exchange of carboxylic acid ligands with CdS nanocrystals.

    PubMed

    Li, Xin; Nichols, Valerie M; Zhou, Dapeng; Lim, Cynthia; Pau, George Shu Heng; Bardeen, Christopher J; Tang, Ming L

    2014-06-11

    We study ligand exchange between the carboxylic acid group and 5.0 nm oleic-acid capped CdS nanocrystals (NCs) using fluorescence resonance energy transfer (FRET). This is the first measurement of the initial binding events between cadmium chalcogenide NCs and carboxylic acid groups. The binding behavior can be described as an interaction between a ligand with single binding group and a substrate with multiple, identical binding sites. Assuming Poissonian binding statistics, our model fits both steady-state and time-resolved photoluminescence (SSPL and TRPL, respectively) data well. A modified Langmuir isotherm reveals that a CdS nanoparticle has an average of 3.0 new carboxylic acid ligands and binding constant, Ka, of 3.4 × 10(5) M(-1).

  13. Acid-Sulfate-Weathering Activity in Shergottite Sites on Mars Recorded in Grim Glasses

    NASA Technical Reports Server (NTRS)

    Rao, M. N.; Nyquist, L. E.; Ross, K.; Sutton, S. R.; Schwandt, C. S.

    2011-01-01

    Based on mass spectrometric studies of sulfur species in Shergotty and EET79001, [1] and [2] showed that sulfates and sulfides occur in different proportions in shergottites. Sulfur speciation studies in gas-rich impact-melt (GRIM) glasses in EET79001 by the XANES method [3] showed that S K-XANES spectra in GRIM glasses from Lith A indicate that S is associated with Ca and Al presumably as sulfides/sulfates whereas the XANES spectra of amorphous sulfide globules in GRIM glasses from Lith B indicate that S is associated with Fe as FeS. In these amorphous iron sulfide globules, [4] found no Ni using FE-SEM and suggested that the globules resulting from immiscible sulfide melt may not be related to the igneous iron sulfides having approximately 1-3% Ni. Furthermore, in the amorphous iron sulfides from 507 GRIM glass, [5] determined delta(sup 34)S values ranging from +3.5%o to -3.1%o using Nano-SIMS. These values plot between the delta(sup 34)S value of +5.25%o determined in the sulfate fraction in Shergotty [6] at one extreme and the value of -1.7%o obtained for igneous sulfides in EET79001 and Shergotty [7] at the other. These results suggest that the amorphous Fe-S globules likely originated by shock reduction of secondary iron sulfate phases occurring in the regolith precursor materials during impact [7]. Sulfates in the regolith materials near the basaltic shergottite sites on Mars owe their origin to surficial acid-sulfate interactions. We examine the nature of these reactions by studying the composition of the end products in altered regolith materials. For the parent material composition, we use that of the host shergottite material in which the impact glasses are situated.

  14. Effects of acidic recharge on groundwater at the St. Kevin Gulch site, Leadville, Colorado

    USGS Publications Warehouse

    Paschke, S.S.; Harrison, W.J.; Walton-Day, K.

    2001-01-01

    The acid rock drainage-affected stream of St. Kevin Gulch recharges the Quaternary sand and gravel aquifer of Tennessee Park, near Leadville, Colorado, lowering pH and contributing iron, cadmium, copper, zinc and sulphate to the ground-water system. Dissolved metal mobility is controlled by the seasonal spring runoff as well as oxidation/reduction (redox) reactions in the aquifer. Oxidizing conditions occur in the unconfined portions of the aquifer whilst sulphate-reducing conditions are found down gradient where semi-confined groundwater flow occurs beneath a natural wetland. Iron-reducing conditions occur in the transition from unconfined to semi-confined groundwater flow. Dissolved iron concentrations are low to not detectable in the alluvial fan recharge zone and increase in a down gradient direction. The effects of low-pH, metal-rich recharge are pronounced during low-flow in the fall when there is a defined area of low pH groundwater with elevated concentrations of dissolved zinc, cadmium, copper and sulphate adjacent to St. Kevin Gulch. Dissolved metal and sulphate concentrations in the recharge zone are diluted during spring runoff, although the maximum concentrations of dissolved zinc, cadmium, copper and sulphate occur at selected down gradient locations during high flow. Dissolved zinc, cadmium and copper concentrations are low to not detectable, whereas dissolved iron concentrations are greatest, in groundwater samples from the sulphate-reducing zone. Attenuation of zinc, cadmium and copper beneath the wetland suggests sulphide mineral precipitation is occurring in the semi-confined aquifer, in agreement with previous site investigations and saturation index calculations. Adsorption of dissolved zinc, cadmium and copper onto iron hydroxides is a minor attenuation process due to the low pH of the groundwater system.

  15. Trace metal biogeochemistry in mangrove ecosystems: a comparative assessment of acidified (by acid sulfate soils) and non-acidified sites.

    PubMed

    Nath, Bibhash; Birch, Gavin; Chaudhuri, Punarbasu

    2013-10-01

    The generation of acidity and subsequent mobilization of toxic metals induced by acid sulfate soils (ASSs) are known to cause severe environmental damage to many coastal wetlands and estuaries of Australia and worldwide. Mangrove ecosystems serve to protect coastal environments, but are increasingly threatened from such ASS-induced acidification due to variable hydrological conditions (i.e., inundation-desiccation cycles). However, the impact of such behaviors on trace metal distribution, bio-availability and accumulation in mangrove tissues, i.e., leaves and pneumatophores, are largely unknown. In this study, we examined how ASS-induced acidifications controlled trace metal distribution and bio-availability in gray mangrove (Avicennia marina) soils and in tissues in the Kooragang wetland, New South Wales, Australia. We collected mangrove soils, leaves and pneumatophores from a part of the wetland acidified from ASS (i.e., an affected site) for detailed biogeochemical studies. The results were compared with samples collected from a natural intertidal mangrove forest (i.e., a control site) located within the same wetland. Soil pH (mean: 5.90) indicated acidic conditions in the affected site, whereas pH was near-neutral (mean: 7.17) in the control site. The results did not show statistically significant differences in near-total and bio-available metal concentrations, except for Fe and Mn, between affected and control sites. Iron concentrations were significantly (p values≤0.001) greater in the affected site, whereas Mn concentrations were significantly (p values≤0.001) greater in the control site. However, large proportions of near-total metals were potentially bio-available in control sites. Concentrations of Fe and Ni were significantly (p values≤0.001) greater in leaves and pneumatophores of the affected sites, whereas Mn, Cu, Pb and Zn were greater in control sites. The degree of metal bio-accumulation in leaves and pneumatophores suggest contrasting

  16. Trace metal biogeochemistry in mangrove ecosystems: a comparative assessment of acidified (by acid sulfate soils) and non-acidified sites.

    PubMed

    Nath, Bibhash; Birch, Gavin; Chaudhuri, Punarbasu

    2013-10-01

    The generation of acidity and subsequent mobilization of toxic metals induced by acid sulfate soils (ASSs) are known to cause severe environmental damage to many coastal wetlands and estuaries of Australia and worldwide. Mangrove ecosystems serve to protect coastal environments, but are increasingly threatened from such ASS-induced acidification due to variable hydrological conditions (i.e., inundation-desiccation cycles). However, the impact of such behaviors on trace metal distribution, bio-availability and accumulation in mangrove tissues, i.e., leaves and pneumatophores, are largely unknown. In this study, we examined how ASS-induced acidifications controlled trace metal distribution and bio-availability in gray mangrove (Avicennia marina) soils and in tissues in the Kooragang wetland, New South Wales, Australia. We collected mangrove soils, leaves and pneumatophores from a part of the wetland acidified from ASS (i.e., an affected site) for detailed biogeochemical studies. The results were compared with samples collected from a natural intertidal mangrove forest (i.e., a control site) located within the same wetland. Soil pH (mean: 5.90) indicated acidic conditions in the affected site, whereas pH was near-neutral (mean: 7.17) in the control site. The results did not show statistically significant differences in near-total and bio-available metal concentrations, except for Fe and Mn, between affected and control sites. Iron concentrations were significantly (p values≤0.001) greater in the affected site, whereas Mn concentrations were significantly (p values≤0.001) greater in the control site. However, large proportions of near-total metals were potentially bio-available in control sites. Concentrations of Fe and Ni were significantly (p values≤0.001) greater in leaves and pneumatophores of the affected sites, whereas Mn, Cu, Pb and Zn were greater in control sites. The degree of metal bio-accumulation in leaves and pneumatophores suggest contrasting

  17. Screening and assessment of solidification/stabilization amendments suitable for soils of lead-acid battery contaminated site.

    PubMed

    Zhang, Zhuo; Guo, Guanlin; Teng, Yanguo; Wang, Jinsheng; Rhee, Jae Seong; Wang, Sen; Li, Fasheng

    2015-05-15

    Lead exposure via ingestion of soil and dust generally occurs at lead-acid battery manufacturing and recycling sites. Screening solidification/stabilization (S/S) amendments suitable for lead contaminated soil in an abandoned lead-acid battery factory site was conducted based on its chemical forms and environmental risks. Twelve amendments were used to immobilize the Pb in soil and assess the solidification/stabilization efficiency by toxicity leaching tests. The results indicated that three amendments, KH₂PO₄ (KP), KH₂PO₄:oyster shell power=1:1 (by mass ratio; SPP), and KH₂PO₄:sintered magnesia=1:1 (by mass ratio; KPM) had higher remediation efficiencies that led to a 92% reduction in leachable Pb with the addition of 5% amendments, while the acid soluble fraction of Pb (AS-Pb) decreased by 41-46% and the residual fraction (RS-Pb) increased by 16-25%. The S/S costs of the three selected amendments KP, SPP, and KPM could be controlled to $22.3 per ton of soil when the Pb concentration in soil ranged from 2000 to 3000 mg/kg. The results of this study demonstrated that KP, SPP, and KPM can effectively decrease bioavailability of Pb. These findings could provide basis for decision-making of S/S remediation of lead-acid battery contaminated sites.

  18. Screening and assessment of solidification/stabilization amendments suitable for soils of lead-acid battery contaminated site.

    PubMed

    Zhang, Zhuo; Guo, Guanlin; Teng, Yanguo; Wang, Jinsheng; Rhee, Jae Seong; Wang, Sen; Li, Fasheng

    2015-05-15

    Lead exposure via ingestion of soil and dust generally occurs at lead-acid battery manufacturing and recycling sites. Screening solidification/stabilization (S/S) amendments suitable for lead contaminated soil in an abandoned lead-acid battery factory site was conducted based on its chemical forms and environmental risks. Twelve amendments were used to immobilize the Pb in soil and assess the solidification/stabilization efficiency by toxicity leaching tests. The results indicated that three amendments, KH₂PO₄ (KP), KH₂PO₄:oyster shell power=1:1 (by mass ratio; SPP), and KH₂PO₄:sintered magnesia=1:1 (by mass ratio; KPM) had higher remediation efficiencies that led to a 92% reduction in leachable Pb with the addition of 5% amendments, while the acid soluble fraction of Pb (AS-Pb) decreased by 41-46% and the residual fraction (RS-Pb) increased by 16-25%. The S/S costs of the three selected amendments KP, SPP, and KPM could be controlled to $22.3 per ton of soil when the Pb concentration in soil ranged from 2000 to 3000 mg/kg. The results of this study demonstrated that KP, SPP, and KPM can effectively decrease bioavailability of Pb. These findings could provide basis for decision-making of S/S remediation of lead-acid battery contaminated sites. PMID:25699676

  19. Site-specific regulation of adult neurogenesis by dietary fatty acid content, vitamin E and flight exercise in European starlings.

    PubMed

    Hall, Zachary J; Bauchinger, Ulf; Gerson, Alexander R; Price, Edwin R; Langlois, Lillie A; Boyles, Michelle; Pierce, Barbara; McWilliams, Scott R; Sherry, David F; Macdougall-Shackleton, Scott A

    2014-03-01

    Exercise is known to have a strong effect on neuroproliferation in mammals ranging from rodents to humans. Recent studies have also shown that fatty acids and other dietary supplements can cause an upregulation of neurogenesis. It is not known, however, how exercise and diet interact in their effects on adult neurogenesis. We examined neuronal recruitment in multiple telencephalic sites in adult male European starlings (Sturnus vulgaris) exposed to a factorial combination of flight exercise, dietary fatty acids and antioxidants. Experimental birds were flown in a wind tunnel following a training regime that mimicked the bird's natural flight behaviour. In addition to flight exercise, we manipulated the composition of dietary fatty acids and the level of enrichment with vitamin E, an antioxidant reported to enhance neuronal recruitment. We found that all three factors - flight exercise, fatty acid composition and vitamin E enrichment - regulate neuronal recruitment in a site-specific manner. We also found a robust interaction between flight training and vitamin E enrichment at multiple sites of neuronal recruitment. Specifically, flight training was found to enhance neuronal recruitment across the telencephalon, but only in birds fed a diet with a low level of vitamin E. Conversely, dietary enrichment with vitamin E upregulated neuronal recruitment, but only in birds not flown in the wind tunnel. These findings indicate conserved modulation of adult neurogenesis by exercise and diet across vertebrate taxa and indicate possible therapeutic interventions in disorders characterized by reduced adult neurogenesis.

  20. Distribution and integrated assessment of lead in an abandoned lead-acid battery site in Southwest China before redevelopment.

    PubMed

    Wang, Mei; Zhang, Chao; Zhang, Zhuo; Li, Fasheng; Guo, Guanlin

    2016-06-01

    Lead-acid battery sites have contributed enormous amounts of lead to the environment, significantly affecting its global biogeochemical cycle and leaving the potential risks to human health. An abandoned lead-acid battery site prepared for redevelopment was selected in order to study the distribution of lead in soils, plants, rhizosphere soils and soil solutions. In total, 197 samples from 77 boreholes were collected and analyzed. Single extractions by acetic acid (HOAc) were conducted to assess the bioavailability and speciation of lead in soils for comparison with the parts of the plants that are aboveground. Health risks for future residential development were evaluated by the integrated exposure uptake biokinetic (IEUBK) model. The results indicated that lead concentrations in 83% of the soil samples exceeded the Chinese Environmental Quality Standard for soil (350 mg/kg for Pb) and mainly occurred at depths between 0 and 1.5 m while accumulating at the surface of demolished construction waste and miscellaneous fill. Lead concentrations in soil solutions and HOAc extraction leachates were linked closely to the contents of aboveground Broussonetia papyrifera and Artemisia annua, two main types of local plants that were found at the site. The probability density of lead in blood (PbB) in excess of 10 µg/dL could overtake the 99% mark in the residential scenario. The findings provided a relatively integrated method to illustrate the onsite investigations and assessment for similar sites before remediation and future development from more comprehensive aspects.

  1. Distribution and integrated assessment of lead in an abandoned lead-acid battery site in Southwest China before redevelopment.

    PubMed

    Wang, Mei; Zhang, Chao; Zhang, Zhuo; Li, Fasheng; Guo, Guanlin

    2016-06-01

    Lead-acid battery sites have contributed enormous amounts of lead to the environment, significantly affecting its global biogeochemical cycle and leaving the potential risks to human health. An abandoned lead-acid battery site prepared for redevelopment was selected in order to study the distribution of lead in soils, plants, rhizosphere soils and soil solutions. In total, 197 samples from 77 boreholes were collected and analyzed. Single extractions by acetic acid (HOAc) were conducted to assess the bioavailability and speciation of lead in soils for comparison with the parts of the plants that are aboveground. Health risks for future residential development were evaluated by the integrated exposure uptake biokinetic (IEUBK) model. The results indicated that lead concentrations in 83% of the soil samples exceeded the Chinese Environmental Quality Standard for soil (350 mg/kg for Pb) and mainly occurred at depths between 0 and 1.5 m while accumulating at the surface of demolished construction waste and miscellaneous fill. Lead concentrations in soil solutions and HOAc extraction leachates were linked closely to the contents of aboveground Broussonetia papyrifera and Artemisia annua, two main types of local plants that were found at the site. The probability density of lead in blood (PbB) in excess of 10 µg/dL could overtake the 99% mark in the residential scenario. The findings provided a relatively integrated method to illustrate the onsite investigations and assessment for similar sites before remediation and future development from more comprehensive aspects. PMID:26921546

  2. A Mutational Analysis of the Active Site Loop Residues in cis-3-Chloroacrylic Acid Dehalogenase

    PubMed Central

    Schroeder, Gottfried K.; Huddleston, Jamison P.; Johnson, William H.; Whitman, Christian P.

    2013-01-01

    cis -3-Chloroacrylic acid dehalogenase (cis-CaaD) from Pseudomonas pavonaceae 170 and a homologue from Corynebacterium glutamicum designated Cg10062 share 34% sequence identity (54% similarity). The former catalyzes a key step in a bacterial catabolic pathway for the nematocide 1,3-dichloropropene, whereas the latter has no known biological activity. Although Cg10062 has the six active site residues (Pro-1, His-28, Arg-70, Arg-73, Tyr-103, Glu-114) that are critical for cis-CaaD activity, it shows only a low level cis-CaaD activity and lacks the specificity of cis-CaaD: Cg10062 processes both isomers of 3-chloroacrylate with a preference for the cis-isomer. Although the basis for these differences is unknown, a comparison of the crystal structures of the enzymes covalently modified by an adduct resulting from their incubation with the same inhibitor offers a possible explanation. A 6-residue active site loop in cis-CaaD shows a strikingly different conformation from that observed in Cg10062: the loop closes down on the active site of cis-CaaD, but not on that of Cg10062. In order to examine what this loop might contribute to cis-CaaD catalysis and specificity, the residues were changed individually to those found in Cg10062. Subsequent kinetic and mechanistic analysis suggests that the T34A mutant of cis-CaaD is more Cg10062-like. The mutant enzyme shows a 4-fold increase in Km (using cis-3-bromoacrylate), but not to the degree observed for Cg10062 (687-fold). The mutation also causes a 4-fold decrease in the burst rate (compared to the wild type cis-CaaD), whereas Cg10062 shows no burst rate. More telling is the reaction of the T34A mutant of cis-CaaD with the alternate substrate, 2,3-butadienoate. In the presence of NaBH4 and the allene, cis-CaaD is completely inactivated after one turnover due to the covalent modification of Pro-1. The same experiment with Cg10062 does not result in the covalent modification of Pro-1. The different outcomes are attributed to

  3. Rapid atomic layer deposition of silica nanolaminates: synergistic catalysis of Lewis/Brønsted acid sites and interfacial interactions

    NASA Astrophysics Data System (ADS)

    Fang, Guoyong; Ma, Jing

    2013-11-01

    Rapid atomic layer deposition (RALD) has been applied to prepare various nanolaminates with repeated multilayer structures. The possible reaction pathways for RALD of the Al2O3/SiO2 nanolaminate using trimethylaluminum (TMA) and tris(tert-butoxy)silanol (TBS) are investigated by using density functional theory (DFT) calculations. The introduction of a Lewis-acid catalyst, TMA, can result in the formation of the catalytic site, which accelerates the propagation of the siloxane polymer. The rate-determining step of whole RALD is the elimination of isobutene of the tert-butoxy groups. The Brønsted acid site of [AlO4] can catalyze the elimination of isobutene. At the same time, the interfacial interactions, such as hydrogen bonding interactions between tert-butoxy groups and the surface, further catalyze the elimination of isobutene and accelerate SiO2 RALD reactions. The synergistic catalysis of Lewis/Brønsted acid sites and interfacial interactions may be applied in the RALD fabrication of other silica nanolaminates, such as HfO2/SiO2, ZrO2/SiO2, and TiO2/SiO2, in microelectronics, catalysis, energy storage, and conversion.Rapid atomic layer deposition (RALD) has been applied to prepare various nanolaminates with repeated multilayer structures. The possible reaction pathways for RALD of the Al2O3/SiO2 nanolaminate using trimethylaluminum (TMA) and tris(tert-butoxy)silanol (TBS) are investigated by using density functional theory (DFT) calculations. The introduction of a Lewis-acid catalyst, TMA, can result in the formation of the catalytic site, which accelerates the propagation of the siloxane polymer. The rate-determining step of whole RALD is the elimination of isobutene of the tert-butoxy groups. The Brønsted acid site of [AlO4] can catalyze the elimination of isobutene. At the same time, the interfacial interactions, such as hydrogen bonding interactions between tert-butoxy groups and the surface, further catalyze the elimination of isobutene and accelerate

  4. Developing palaeolimnological records of organic content (DOC and POC) using the UK Acid Water Monitoring Network sites

    NASA Astrophysics Data System (ADS)

    Russell, Fiona; Chiverrell, Richard; Boyle, John

    2016-04-01

    Monitoring programmes have shown increases in concentrations of dissolved organic matter (DOM) in the surface waters of northern and central Europe (Monteith et al. 2007), and negative impacts of the browning of river waters have been reported for fish populations (Jonsson et al. 2012; Ranaker et al. 2012) and for ecosystem services such as water treatment (Tuvendal and Elmqvist 2011). Still the exact causes of the recent browning remain uncertain, the main contenders being climate change (Evans et al. 2005) and reduced ionic strength in surface water resulting from declines in anthropogenic sulphur and sea salt deposition (Monteith et al. 2007). There is a need to better understand the pattern, drivers and trajectory of these increases in DOC and POC in both recent and longer-term (Holocene) contexts to improve the understanding of carbon cycling within lakes and their catchments. In Britain there are some ideal sites for testing whether these trends are preserved and developing methods for reconstructing organic fluxes from lake sedimentary archives. There is a suite of lakes distributed across the country, the UK Acid Waters Monitoring Network (UKAWMN) sites, which have been monitored monthly for dissolved organic carbon and other aqueous species since 1988. These 12 lakes have well studied recent and in some case whole Holocene sediment records. Here four of those lakes (Grannoch, Chon, Scoat Tarn and Cwm Mynach) are revisited, with sampling focused on the sediment-water interface and very recent sediments (approx.150 years). At Scoat Tarn (approx. 1000 years) and Llyn Mynach (11.5k years) longer records have been obtained to assess equivalent patterns through the Holocene. Analyses of the gravity cores have focused on measuring and characterising the organic content for comparison with recorded surface water DOC measurements (UKAWMN). Data from pyrolysis measurements (TGA/DSC) in an N atmosphere show that the mass loss between 330-415°C correlates well with

  5. Strength Testing.

    ERIC Educational Resources Information Center

    Londeree, Ben R.

    1981-01-01

    Postural deviations resulting from strength and flexibility imbalances include swayback, scoliosis, and rounded shoulders. Screening tests are one method for identifying strength problems. Tests for the evaluation of postural problems are described, and exercises are presented for the strengthening of muscles. (JN)

  6. Hyperfine and Nuclear Quadrupole Tensors of Nitrogen Donors in the QA Site of Bacterial Reaction Centers: Correlation of the Histidine Nδ Tensors with Hydrogen Bond Strength

    PubMed Central

    2015-01-01

    X- and Q-band pulsed EPR spectroscopy was applied to study the interaction of the QA site semiquinone (SQA) with nitrogens from the local protein environment in natural abundance 14N and in 15N uniformly labeled photosynthetic reaction centers of Rhodobacter sphaeroides. The hyperfine and nuclear quadrupole tensors for His-M219 Nδ and Ala-M260 peptide nitrogen (Np) were estimated through simultaneous simulation of the Q-band 15N Davies ENDOR, X- and Q-band 14,15N HYSCORE, and X-band 14N three-pulse ESEEM spectra, with support from DFT calculations. The hyperfine coupling constants were found to be a(14N) = 2.3 MHz, T = 0.3 MHz for His-M219 Nδ and a(14N) = 2.6 MHz, T = 0.3 MHz for Ala-M260 Np. Despite that His-M219 Nδ is established as the stronger of the two H-bond donors, Ala-M260 Np is found to have the larger value of a(14N). The nuclear quadrupole coupling constants were estimated as e2Qq/4h = 0.38 MHz, η = 0.97 and e2Qq/4h = 0.74 MHz, η = 0.59 for His-M219 Nδ and Ala-M260 Np, respectively. An analysis of the available data on nuclear quadrupole tensors for imidazole nitrogens found in semiquinone-binding proteins and copper complexes reveals these systems share similar electron occupancies of the protonated nitrogen orbitals. By applying the Townes–Dailey model, developed previously for copper complexes, to the semiquinones, we find the asymmetry parameter η to be a sensitive probe of the histidine Nδ–semiquinone hydrogen bond strength. This is supported by a strong correlation observed between η and the isotropic coupling constant a(14N) and is consistent with previous computational works and our own semiquinone-histidine model calculations. The empirical relationship presented here for a(14N) and η will provide an important structural characterization tool in future studies of semiquinone-binding proteins. PMID:25026433

  7. High strength of physical hydrogels based on poly(acrylic acid)-g-poly(ethylene glycol) methyl ether: role of chain architecture on hydrogel properties.

    PubMed

    Yang, Jun; Gong, Cheng; Shi, Fu-Kuan; Xie, Xu-Ming

    2012-10-01

    This investigation was to study the connections between polymer branch architecture of physical hydrogels and their properties. The bottle-brush-like polymer chains of poly(acrylic acid)-g-poly(ethylene glycol) methyl ether (PAA-g-mPEG) with PAA as backbones and mPEG as branch architecture were synthesized and in situ grafted from silica nanoparticles (SNs) to construct hydrogels cross-linked networks in aqueous solutions. The structural variables to be discussed included molecular weight and molar ratio of branch chains, and new aspects of the formation mechanism of physical hydrogels with branch structure in the absence of organic cross-links were present. The results indicated that the differences of polymer chain architecture could be distinguished via their different interactions that are present by gelation process and mature gel properties, such as gel strength and swelling ratio. The gelation occurred at the critical polymer concentration and molecular weight, respectively, and the inorganic/organic (SNs/PAA-g-mPEG) nanoparticles began to entangle and construct the cross-linking networks afterward. The gel-to-sol transition temperature (T(g-s)) and radii of SNs that were encapsulated by polymer chains as a function of time for chains' disentanglement were monitored according to the observation of the dissolution process, and the molecular weight between two consecutive entanglements (M(e)) was calculated thereafter. This study showed that the introduction of branch chain onto the linear backbone significantly promoted the chain interactions and increased entanglement density, which contributed to the hydrogels' network integrity and rigidity, thus illustrating greater elongation at break and tensile strength than the hydrogels formulated with linear polymer chains.

  8. Orchidectomy-induced alterations in volumetric bone density, cortical porosity and strength of femur are attenuated by dietary conjugated linoleic acid in aged guinea pigs.

    PubMed

    DeGuire, Jason R; Mak, Ivy L; Lavery, Paula; Agellon, Sherry; Wykes, Linda J; Weiler, Hope A

    2015-04-01

    Age-related osteoporosis and sarcopenia are ascribed in part to reductions in anabolic hormones. Dietary conjugated linoleic acid (CLA) improves lean and bone mass, but its impact during androgen deficiency is not known. This study tested if CLA would attenuate the effects of orchidectomy (ORX)-induced losses of bone and lean tissue. Male guinea pigs (n=40; 70-72 weeks), were randomized into four groups: (1) SHAM+Control diet, (2) SHAM+CLA diet, (3) ORX+Control diet, (4) ORX+CLA diet. Baseline blood sampling and dual-energy X-ray absorptiometry (DXA) scans were conducted, followed by surgery 4 days later with the test diets started 7 days after baseline sampling. Serial blood sampling and DXA scans were repeated 2, 4, 8 and 16 weeks on the test diets. Body composition and areal BMD (aBMD) of whole body, lumbar spine, femur and tibia were measured using DXA. At week 16, muscle protein fractional synthesis rate (FSR), volumetric BMD (vBMD), microarchitecture and bone strength were assessed. Body weight declined after SHAM and ORX surgery, with slower recovery in the ORX group. Dietary CLA did not affect weight or lean mass, but attenuated gains in fat mass. Lean mass was stable in SHAM and reduced in ORX by 2 weeks with whole body and femur bone mineral content (BMC) reduced by 4 weeks; CLA did not alter BMC. By week 16 ORX groups had lower free testosterone and myofibrillar FSR, yet higher cortisol, osteocalcin and ionized calcium with no alterations due to CLA. ORX+Control had higher prostaglandin E2 (PGE2) and total alkaline phosphatase compared to SHAM+Control whereas ORX+CLA were not different from SHAM groups. Femur metaphyseal vBMD was reduced in ORX+CTRL with the reduction attenuated by CLA. Femur cortical thickness (Ct.Th.) and biomechanical strength were reduced and cortical porosity (Ct.Po.) elevated by ORX and attenuated by CLA. This androgen deficient model with a sarcopenic-osteoporotic phenotype similar to aging men responded to dietary CLA with

  9. Methyl Acetate Synthesis by Esterification on the Modified Ferrierite: Correlation of Acid Sites Measured by Pyridine IR and NH3-TPD for Steady-State Activity.

    PubMed

    Park, Jae Hyun; Pang, Changhyun; Chung, Chan-Hwa; Bae, Jong Wook

    2016-05-01

    The amounts of Brønsted acid sites on K, P, and Zr-modified microporous Ferrierite zeolite were investigated through pyridine FT-IR and NH3-TPD analyses. P-modified Ferrierite showed a superior catalytic activity for methyl acetate synthesis by esterification of methanol and acetic acid. The catalytic activity at steady-state with the acidic properties of as-prepared catalysts was well correlated with the results of pyridine FT-IR (intensity ratio of Brønsted acid sites to total acid sites) compared with that of NH3-TPD. The results can suggest the proper and simple method to estimate the esterification activity at steady-state using the measured acid sites on the as-prepared zeolites. PMID:27483801

  10. Site-specific incorporation of redox active amino acids into proteins

    DOEpatents

    Alfonta, Lital; Schultz, Peter G.; Zhang, Zhiwen

    2009-02-24

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate redox active amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with redox active amino acids using these orthogonal pairs.

  11. Site-specific incorporation of redox active amino acids into proteins

    DOEpatents

    Alfonta; Lital , Schultz; Peter G. , Zhang; Zhiwen

    2010-10-12

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate redox active amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with redox active amino acids using these orthogonal pairs.

  12. Site-specific incorporation of redox active amino acids into proteins

    SciTech Connect

    Alfonta, Lital; Schultz, Peter G.; Zhang, Zhiwen

    2012-02-14

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate redox active amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with redox active amino acids using these orthogonal pairs.

  13. Site-specific incorporation of redox active amino acids into proteins

    SciTech Connect

    Alfonta, Lital; Schultz, Peter G.; Zhang, Zhiwen

    2011-08-30

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate redox active amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with redox active amino acids using these orthogonal pairs.

  14. Coupling acidic organelles with the ER through Ca²⁺ microdomains at membrane contact sites.

    PubMed

    Penny, Christopher J; Kilpatrick, Bethan S; Eden, Emily R; Patel, Sandip

    2015-10-01

    Acidic organelles such as lysosomes serve as non-canonical Ca(2+) stores. The Ca(2+) mobilising messenger NAADP is thought to trigger local Ca(2+) release from such stores. These events are then amplified by Ca(2+) channels on canonical ER Ca(2+) stores to generate physiologically relevant global Ca(2+) signals. Coupling likely occurs at microdomains formed at membrane contact sites between acidic organelles and the ER. Molecular analyses and computational modelling suggest heterogeneity in the composition of these contacts and predicted Ca(2+) microdomain behaviour. Conversely, acidic organelles might also locally amplify and temper ER-evoked Ca(2+) signals. Ca(2+) microdomains between distinct Ca(2+) stores are thus likely to be integral to the genesis of complex Ca(2+) signals. PMID:25866010

  15. Role of arginine 285 in the active site of Rhodotorula gracilis D-amino acid oxidase. A site-directed mutagenesis study.

    PubMed

    Molla, G; Porrini, D; Job, V; Motteran, L; Vegezzi, C; Campaner, S; Pilone, M S; Pollegioni, L

    2000-08-11

    Arg(285), one of the very few conserved residues in the active site of d-amino acid oxidases, has been mutated to lysine, glutamine, aspartate, and alanine in the enzyme from the yeast Rhodotorula gracilis (RgDAAO). The mutated proteins are all catalytically competent. Mutations of Arg(285) result in an increase ( approximately 300-fold) of K(m) for the d-amino acid and in a large decrease ( approximately 500-fold) of turnover number. Stopped-flow analysis shows that the decrease in turnover is paralleled by a similar decrease in the rate of flavin reduction (k(2)), the latter still being the rate-limiting step of the reaction. In agreement with data from the protein crystal structure, loss of the guanidinium group of Arg(285) in the mutated DAAOs drastically reduces the binding of several carboxylic acids (e.g. benzoate). These results highlight the importance of this active site residue in the precise substrate orientation, a main factor in this redox reaction. Furthermore, Arg(285) DAAO mutants have spectral properties similar to those of the wild-type enzyme, but show a low degree of stabilization of the flavin semiquinone and a change in the redox properties of the free enzyme. From this, we can unexpectedly conclude that Arg(285) in the free enzyme form is involved in the stabilization of the negative charge on the N(1)-C(2)=O locus of the isoalloxazine ring of the flavin. We also suggest that the residue undergoes a conformational change in order to bind the carboxylate portion of the substrate/ligand in the complexed enzyme. PMID:10821840

  16. Novel human D-amino acid oxidase inhibitors stabilize an active-site lid-open conformation

    PubMed Central

    Terry-Lorenzo, Ryan T.; Chun, Lawrence E.; Brown, Scott P.; Heffernan, Michele L. R.; Fang, Q. Kevin; Orsini, Michael A.; Pollegioni, Loredano; Hardy, Larry W.; Spear, Kerry L.; Large, Thomas H.

    2014-01-01

    The NMDAR (N-methyl-D-aspartate receptor) is a central regulator of synaptic plasticity and learning and memory. hDAAO (human D-amino acid oxidase) indirectly reduces NMDAR activity by degrading the NMDAR co-agonist D-serine. Since NMDAR hypofunction is thought to be a foundational defect in schizophrenia, hDAAO inhibitors have potential as treatments for schizophrenia and other nervous system disorders. Here, we sought to identify novel chemicals that inhibit hDAAO activity. We used computational tools to design a focused, purchasable library of compounds. After screening this library for hDAAO inhibition, we identified the structurally novel compound, ‘compound 2’ [3-(7-hydroxy-2-oxo-4-phenyl-2H-chromen-6-yl)propanoic acid], which displayed low nM hDAAO inhibitory potency (Ki=7 nM). Although the library was expected to enrich for compounds that were competitive for both D-serine and FAD, compound 2 actually was FAD uncompetitive, much like canonical hDAAO inhibitors such as benzoic acid. Compound 2 and an analog were independently co-crystalized with hDAAO. These compounds stabilized a novel conformation of hDAAO in which the active-site lid was in an open position. These results confirm previous hypotheses regarding active-site lid flexibility of mammalian D-amino acid oxidases and could assist in the design of the next generation of hDAAO inhibitors. PMID:25001371

  17. The Effectiveness of Predict-Observe-Explain Technique in Probing Students' Understanding about Acid-Base Chemistry: A Case for the Concepts of pH, pOH, and Strength

    ERIC Educational Resources Information Center

    Kala, Nesli; Yaman, Fatma; Ayas, Alipasa

    2013-01-01

    The present study describes high school students' conceptions about acids and bases in terms of pH, pOH, microscopic level, strength, and concentration. A total of 27 high school students participated in the study. The data was collected using 3 POE tasks and a semi-structured interview. The data analysis demonstrated that most of the students had…

  18. Site-directed gene mutation at mixed sequence targets by psoralen-conjugated pseudo-complementary peptide nucleic acids

    PubMed Central

    Kim, Ki-Hyun; Nielsen, Peter E.; Glazer, Peter M.

    2007-01-01

    Sequence-specific DNA-binding molecules such as triple helix-forming oligonucleotides (TFOs) provide a means for inducing site-specific mutagenesis and recombination at chromosomal sites in mammalian cells. However, the utility of TFOs is limited by the requirement for homopurine stretches in the target duplex DNA. Here, we report the use of pseudo-complementary peptide nucleic acids (pcPNAs) for intracellular gene targeting at mixed sequence sites. Due to steric hindrance, pcPNAs are unable to form pcPNA–pcPNA duplexes but can bind to complementary DNA sequences by Watson–Crick pairing via double duplex-invasion complex formation. We show that psoralen-conjugated pcPNAs can deliver site-specific photoadducts and mediate targeted gene modification within both episomal and chromosomal DNA in mammalian cells without detectable off-target effects. Most of the induced psoralen-pcPNA mutations were single-base substitutions and deletions at the predicted pcPNA-binding sites. The pcPNA-directed mutagenesis was found to be dependent on PNA concentration and UVA dose and required matched pairs of pcPNAs. Neither of the individual pcPNAs alone had any effect nor did complementary PNA pairs of the same sequence. These results identify pcPNAs as new tools for site-specific gene modification in mammalian cells without purine sequence restriction, thereby providing a general strategy for designing gene targeting molecules. PMID:17977869

  19. Conceptual Model of Uranium in the Vadose Zone for Acidic and Alkaline Wastes Discharged at the Hanford Site Central Plateau

    SciTech Connect

    Truex, Michael J.; Szecsody, James E.; Qafoku, Nikolla; Serne, R. Jeffrey

    2014-09-01

    Historically, uranium was disposed in waste solutions of varying waste chemistry at the Hanford Site Central Plateau. The character of how uranium was distributed in the vadose zone during disposal, how it has continued to migrate through the vadose zone, and the magnitude of potential impacts on groundwater are strongly influenced by geochemical reactions in the vadose zone. These geochemical reactions can be significantly influenced by the disposed-waste chemistry near the disposal location. This report provides conceptual models and supporting information to describe uranium fate and transport in the vadose zone for both acidic and alkaline wastes discharged at a substantial number of waste sites in the Hanford Site Central Plateau. The conceptual models include consideration of how co-disposed acidic or alkaline fluids influence uranium mobility in terms of induced dissolution/precipitation reactions and changes in uranium sorption with a focus on the conditions near the disposal site. This information, when combined with the extensive information describing uranium fate and transport at near background pH conditions, enables focused characterization to support effective fate and transport estimates for uranium in the subsurface.

  20. Extending the Diffuse Layer Model of Surface Acidity Behavior: III. Estimating Bound Site Activity Coefficients

    EPA Science Inventory

    Although detailed thermodynamic analyses of the 2-pK diffuse layer surface complexation model generally specify bound site activity coefficients for the purpose of accounting for those non-ideal excess free energies contributing to bound site electrochemical potentials, in applic...

  1. Analysis of the five glycosylation sites of human alpha 1-acid glycoprotein.

    PubMed Central

    Treuheit, M J; Costello, C E; Halsall, H B

    1992-01-01

    Orosomucoid (OMD) contains complex bi-, tri- and tetra-antennary glycan chains. Subfractionation of OMD into three molecular variants using concanavalin A lectin chromatography is based on variations in these complex structures. Standard h.p.l.c. profiles have been developed to analyse the percentage and distribution of the glycoforms present at each glycosylation site in OMD and its molecular variants. The ability to quantify the glycoforms present at each site allows us to extend the earlier results of others and resolve the remaining questions concerning the glycan structures of these variants. Most significantly, the proportions of bi-, tri- and tetra-antennary chains differ at each site for the three molecular variants. The most strongly retained variant from concanavalin A is uniquely capable of possessing biantennary chains at all five sites, whereas the unretained variant is completely devoid of biantennary chains. Only glycosylation site II of the five present is 100% biantennary in the retained and weakly retained variants. In addition, the two gene products of OMD were differentially glycosylated. Molecular masses of the glycoforms were verified by matrix-assisted u.v. laser desorption mass spectrometry. On the basis of the site distribution of oligosaccharides in the variants, efforts were made to understand the factors that control the processing of the carbohydrate chains in OMD. The results indicate that the 'site-directed' model of processing offers the most consistent explanation for the structures seen at the individual glycosylation sites of OMD. PMID:1567356

  2. Corrosion inhibition in 2.0 M sulfuric acid solutions of high strength maraging steel by aminophenyl tetrazole as a corrosion inhibitor

    NASA Astrophysics Data System (ADS)

    Sherif, El-Sayed M.

    2014-02-01

    The corrosion of high strength maraging steel after varied immersion times in concentrated solution, 2.0 M, of sulfuric acid has been investigated. The work was also extended to study the effect of 5-(3-aminophenyl)-tetrazole (APTA) on the inhibition of the steel corrosion. The study has been carried out using electrochemical impedance spectroscopy (EIS), potentiodynamic polarization and scanning electron microscope (SEM) along with energy dispersive X-ray analyzer (EDX) investigations. EIS spectra showed that the corrosion and polarization resistances decrease with increasing the immersion time of the steel before measurement and increase in the presence of APTA and the increase of its concentration. Polarization data agreed with the EIS measurements and indicated that the increase of immersion time increases the corrosion of steel by increasing its corrosion current and corrosion rate and lowering its polarization resistance. On the other hand, the addition of APTA and the increase of its concentration minimized the corrosion of steel through decreasing the corrosion current and corrosion rate and increasing the polarization resistance at all exposure test periods. SEM and EDX investigations confirmed that the inhibition of the maraging steel in the 2.0 M H2SO4 solutions is achieved via the adsorption of the APTA molecules onto the steel protecting its surface from being dissolved easily.

  3. Hindered Diffusion of Oligosaccharides in High Strength Poly(ethylene glycol)/Poly(acrylic acid) Interpenetrating Network Hydrogels: Hydrodynamic Versus Obstruction Models

    PubMed Central

    Waters, Dale J.; Frank, Curtis W.

    2010-01-01

    Diffusion coefficients of small oligosaccharides within high strength poly(ethylene glycol)/poly(acrylic acid) interpenetrating network (PEG/PAA IPN) hydrogels were measured by diffusion through hydrogel slabs. The ability of hindered diffusion models previously presented in the literature to fit the experimental data is examined. A model based solely on effects due to hydrodynamics is compared to a model based solely on solute obstruction. To examine the effect of polymer volume fraction on the observed diffusion coefficients, the equilibrium volume fraction of polymer in PEG/PAA IPNs was systematically varied by changing the initial PEG polymer concentration in hydrogel precursor solutions from 20 to 50 wt./wt.%. To examine the effect of solute radius on the observed diffusion coefficients, solute radii were varied from 3.3 to 5.1 Å by measuring diffusion coefficients of glucose, a monosaccharide; maltose, a disaccharide; and maltotriose, a trisaccharide. Both the hydrodynamic and obstruction models rely on scaling relationships to predict diffusion coefficients. The proper scaling relationship for each of the hindered diffusion models is evaluated based on fits to experimental data. The scaling relationship employed is found to have a greater significance for the hydrodynamic model than the obstruction model. Regardless of the scaling relationship employed, the obstruction model provides a better fit to our experimental data than the hydrodynamic model. PMID:20514136

  4. Evaluation of biomechanical strength, stability, bioactivity, and in vivo biocompatibility of a novel calcium deficient hydroxyapatite/poly(amino acid) composite cervical vertebra cage.

    PubMed

    Xiong, Yi; Li, Hong; Zhou, Chunguang; Yang, Xi; Song, Yueming; Qing, Yan; Yan, Yonggang

    2014-01-01

    A new type of cervical vertebra cage was prepared using a novel composite, calcium deficient hydroxyapatite/poly(amino acid) (HA/PAA), and its mechanical properties, in vitro stability and bioactivity, and in vivo biocompatibility were characterized. The results showed that the axial compressive loads of the HA/PAA cage were in the range of 10058-10612 N and the lateral compressive loads were in the range of 1180-2363 N, and varied with the height of the cervical vertebra cages. After immersion in simulated body fluid (SBF) for 16 weeks, the axial compressive loads of the cage decreased from 10058 to 7131 N and the lateral compressive loads decreased from 1180 to 479 N. In addition, the weight loss decreased 6.01%, showing that HA/PAA composites had good stability during the incubation period. The pH value of SBF was also monitored during the whole soaking period; it fluctuated in the range of 6.9-7.4. Scanning electron microscope and energy dispersive spectrometer results showed the cage was bioactive with a new apatite layer attached on the surface. The histological evaluation revealed that new bone tissue bonded tightly with the surfaces of the implants, showing excellent biocompatibility. In conclusion, the HA/PAA cage showed sufficient strength, good stability, bioactivity, and biocompatibility, and has potential applications for clinical cervical vertebrae repair.

  5. Biodegradation of industrial-strength 2,4-dichlorophenoxyacetic acid wastewaters in the presence of glucose in aerobic and anaerobic sequencing batch reactors.

    PubMed

    Elefsiniotis, Panagiotis; Wareham, David G

    2013-01-01

    This research explored the biodegradability of 2,4-dichlorophenoxyacetic acid (2,4-D) in two laboratory-scale sequencing batch reactors (SBRs) that operated under aerobic and anaerobic conditions. The potential limit of 2,4-D degradation was investigated at a hydraulic retention time of 48 h, using glucose as a supplemental substrate and increasing feed concentrations of 2,4-D; namely 100 to 700 mg/L (i.e. industrial strength) for the aerobic system and 100 to 300 mg/L for the anaerobic SBR. The results revealed that 100 mg/L of 2,4-D was completely degraded following an acclimation period of 29 d (aerobic SBR) and 70 d (anaerobic SBR). The aerobic system achieved total 2,4-D removal at feed concentrations up to 600 mg/L which appeared to be a practical limit, since a further increase to 700 mg/L impaired glucose degradation while 2,4-D biodegradation was non-existent. In all cases, glucose was consumed before the onset of 2,4-D degradation. In the anaerobic SBR, 2,4-D degradation was limited to 120 mg/L.

  6. Transport behavior of humic acid-modified nano-hydroxyapatite in saturated packed column: effects of Cu, ionic strength, and ionic composition.

    PubMed

    Wang, Dengjun; Chu, Lingyang; Paradelo, Marcos; Peijnenburg, Willie J G M; Wang, Yujun; Zhou, Dongmei

    2011-08-15

    The surfaces of nano-hydroxyapatite (nHAP) used for contaminated soil and groundwater remediation may be modified to render nHAP highly mobile in the subsurface. Humic acid (HA) is widely used to modify and stabilize colloid suspensions. In this work, column experiments were conducted to determine the effects of contaminant (e.g., Cu) concentration, ionic strength (IS), and ion composition (IC) on the transport behavior of HA-modified nHAP in saturated packed columns. IS and nature of the cation had strong effects on the deposition of nHAP, and the effect was greater for divalent than for monovalent cations. Divalent cations have a greater capacity to screen the surface charge of nHAP, and Ca(2+) bridges the HA-modified nHAP colloidal particles, which causes greater deposition. Moreover, Cu(2+) had a greater effect on the transport behavior than Ca(2+) due to their strong exchange with Ca(2+) of nHAP and its surface complexation with nHAP. The relative travel distance L(T), of the injected HA-modified nHAP colloids, ranges from less than one to several meters at varying Cu concentrations, ISs, and ICs in saturated packed columns. The results are crucial to evaluate the efficacy of nHAP on the remediation of contaminated soil and groundwater environments.

  7. Computational investigation of locked nucleic acid (LNA) nucleotides in the active sites of DNA polymerases by molecular docking simulations.

    PubMed

    Poongavanam, Vasanthanathan; Madala, Praveen K; Højland, Torben; Veedu, Rakesh N

    2014-01-01

    Aptamers constitute a potential class of therapeutic molecules typically selected from a large pool of oligonucleotides against a specific target. With a scope of developing unique shorter aptamers with very high biostability and affinity, locked nucleic acid (LNA) nucleotides have been investigated as a substrate for various polymerases. Various reports showed that some thermophilic B-family DNA polymerases, particularly KOD and Phusion DNA polymerases, accepted LNA-nucleoside 5'-triphosphates as substrates. In this study, we investigated the docking of LNA nucleotides in the active sites of RB69 and KOD DNA polymerases by molecular docking simulations. The study revealed that the incoming LNA-TTP is bound in the active site of the RB69 and KOD DNA polymerases in a manner similar to that seen in the case of dTTP, and with LNA structure, there is no other option than the locked C3'-endo conformation which in fact helps better orienting within the active site. PMID:25036012

  8. Evaluation of Shear Strength Threshold of Concern for Retrieval of Interim-Stored K-Basin Sludge in the Hanford Site

    SciTech Connect

    Onishi, Yasuo; Yokuda, Satoru T.; Schmidt, Andrew J.

    2010-11-01

    K-Basin sludge will be recovered into the Sludge Transport and Storage Containers (STSCs) and will be stored in the T Plant for interim storage (at least 10 years). Long-term sludge storage tests conducted by Pacific Northwest National Laboratory show that high uranium content K Basin sludge can self-cement and form a strong sludge with a bulk shear strength of up to 65 kPa. Some of this sludge has "paste" and "chunks" with shear strengths of approximately 3~5 kPa and 380 ~ 770 kPa, respectively. High uranium content sludge samples subjected to hydrothermal testing (e.g., 185°C, 10 h) have been observed to form agglomerates with a shear strength up to 170 kPa. After interim storage at T Plant, the sludge in the STSCs will be mobilized by water jets impinging the sludge. The objective of the evaluation was to determine the range of sludge shear strength for which there is high confidence that a water-jet retrieval system can mobilize stored K-Basin sludge from STSCs. The shear strength at which the sludge can be retrieved is defined as the "shear strength threshold of concern." If the sludge shear strength is greater than the value of the shear strength threshold of concern, a water-jet retrieval system will be unlikely to mobilize the sludge up to the container’s walls. The shear strength threshold of concern can be compared with the range of possible shear strengths of K-Basin stored sludge to determine if the current post interim-storage, water-jet retrieval method is adequate. Fourteen effective cleaning radius (ECR) models were reviewed, and their validity was examined by applying them to Hanford 241-SY-101 and 241-AZ-101 Tanks to reproduce the measured ECR produced by the mixer pumps. The validation test identified that the Powell-3 and Crowe-2 ECR models are more accurate than other ECR models reviewed. These ECR models were used to address a question as to whether the effective cleaning radius of a water jet is sufficient or if it can be readily expanded

  9. [Identification of rat and human hemoglobin acetilation sites after its interaction with acetylsalicylic acid].

    PubMed

    Shreĭner, E V; Murashko, E A; Dubrovskiĭ, Ia D; Krasnov, N V; Podol'skaia, E P; Babakov, V N

    2012-01-01

    The possibility of interaction of 0.1 mg/mL acetylsalicylic acid with purified human and rat globin in vitro during 24 h at 37 degrees C was investigated. The rat globin can be modified with acetylsalicylic acid on aminoacid residues K-17, K-57, K-91, K-140 in alpha subunit as well as on K-18, K-77 in beta subunit. The human globin can be modified with acetylsalicylic acid on aminoacid residues K-17, K-41, K-57 and K-91 in alpha subunit as well as on K-18, K-96 and K- 133 in beta subunit. We identified of acetetylated lysines K-17 and K-57 in alpha subunit of human hemoglobin after incubation whole blood with 0.1 mg/mL acetylsalicylic acid during 3 h.

  10. The impact of acid treatment on soilwater chemistry at the Humex site

    SciTech Connect

    Vogt, R.D.; Ranneklev, S.B.; Mykkelbost, T.C. )

    1994-01-01

    The effects of acid treatment on soil water dissolved organic carbon (DOC) and inorganic chemistry are being monitored at the Humic Lake Acidification Experiment (HUMEX) in western Norway. The HUMEX project involves artificial acidification of half of a dystrophic lake and the corresponding drainage basin. Soil water chemistry data were collected from 30 monitoring lysimeters and 130 grid lysimeters. The samples from the monitoring lysimeters were collected before and, for a period of two years, after the onset of acid treatment. Operationally-defined functional fractions of DOC showed that hydrophilic (HPI) and hydrophobic (HPO) acids account for 60% to 90% of the DOC. In soils rich in DOC, the HPO acids were dominant, whereas in mineral soil horizons low in DOC, the HPI acid fractions were highest. The amount of DOC relative to labile aluminum and iron may determine the HPO/HPI acid ratio. The sulphate concentration has increased more in the treated than in the reference side. Coincident decreases in DOC and organically complexes aluminum (Al[sub 0]) concentrations were observed for surface histosol locations. The temporal and spatial variations in c(Al[sub 0]) were mainly explained by variation in c(DOC). 26 refs., 5 figs., 3 tabs.

  11. Low-cost silica, calcite and metal sulfide scale control through on-site production of sulfurous acid from H{sub 2}S or elemental sulfur

    SciTech Connect

    Gallup, D.L.; Kitz, K.

    1997-12-31

    UNOCAL Corporation currently utilizes brine pH modification technology to control scale deposition. Acids utilized in commercial operations include, sulfuric and hydrochloric. A new process reduces costs by producing acid on-site by burning hydrogen sulfide or elemental sulfur. Hydrogen sulfide in non-condensible gas emissions is reduced by oxidization to sulfurous acid. Brine or condensate is treated with sulfurous acid to control scale deposition, mitigate corrosion and improve gas partitioning in condensers.

  12. Specificity of the antibody receptor site to D-lysergamide: model of a physiological receptor for lysergic acid diethylamide.

    PubMed

    Van Vunakis, H; Farrow, J T; Gjika, H B; Levine, L

    1971-07-01

    Antibodies to D-lysergic acid have been produced in rabbits and guinea pigs and a radioimmunoassay for the hapten was developed. The specificity of this lysergamide-antilysergamide reaction was determined by competitive binding with unlabeled lysergic acid diethylamide (LSD), psychotomimetic drugs, neurotransmitters, and other compounds with diverse structures. LSD and several related ergot alkaloids were potent competitors, three to seven times more potent than lysergic acid itself. The N,N-dimethyl derivatives of several compounds, including tryptamine, 5-hydroxytryptamine, 4-hydroxytryptamine, 5-methoxytryptamine, tyramine, and mescaline, were only about ten times less effective than lysergic acid, even though these compounds lack some of the ring systems of lysergic acid. The pattern of inhibition by related compounds with various substituents suggests that the antibody receptor site recognizes structural features resembling the LSD molecule. In particular, the aromatic nucleus and the dimethylated ethylamine side chain in phenylethylamine and tryptamine derivatives may assume in solution a conformation resembling ring A and the methylated nitrogen in ring C of LSD. Among the tryptamine derivatives, a large percentage of the most potent competitors are also psychotomimetic compounds.

  13. Runoff of acidic substances that originated from atmospheric deposition on Yakushima Island, a world natural heritage site.

    PubMed

    Nagafuchi, O; Kakimoto, H; Ebise, S; Inoue, T; Koga, M

    2001-01-01

    In this paper we present monitoring data of stream waters that may reflect acidic impacts on the island as well as the rainwater qualities. The pH ranges of the river water in the Kawara streams in the western part of the island and the Yodogo stream in the central part of the island were 5.71-6.35 and 5.85-6.12 during 1992-1999, respectively. The concentrations of SO4(2-) and NO3- in the river water were lower than those in the rainwater. Many differences were observed among the sampling sites. Higher concentrations of acid substances are found in the stream waters of the western area compared to the other areas. On the other hand, sulfuric acid is the major acid in the rainwater, snow and rime ice. No differences were observed in the ion constituents of the rainwater collected in the areas. These results suggested that the densely growing canopy may play a role in holding air pollutants, and acidic substances deposited on the canopy would be discharged as a through-fall and a stem flow. Furthermore, the water mass containing high ionic substances in the western area has been held in the groundwater layer, continuously supplying the stream waters during dry weather days. On the other hand, part of the basic runoff will be diluted with a surface runoff during the rainy days. As a result, the concentrations of the ionic substances in the stream waters during rainy days decreased.

  14. CD44 Binding to Hyaluronic Acid Is Redox Regulated by a Labile Disulfide Bond in the Hyaluronic Acid Binding Site

    PubMed Central

    Kellett-Clarke, Helena; Stegmann, Monika; Barclay, A. Neil; Metcalfe, Clive

    2015-01-01

    CD44 is the primary leukocyte cell surface receptor for hyaluronic acid (HA), a component of the extracellular matrix. Enzymatic post translational cleavage of labile disulfide bonds is a mechanism by which proteins are structurally regulated by imparting an allosteric change and altering activity. We have identified one such disulfide bond in CD44 formed by Cys77 and Cys97 that stabilises the HA binding groove. This bond is labile on the surface of leukocytes treated with chemical and enzymatic reducing agents. Analysis of CD44 crystal structures reveal the disulfide bond to be solvent accessible and in the–LH hook configuration characteristic of labile disulfide bonds. Kinetic trapping and binding experiments on CD44-Fc chimeric proteins show the bond is preferentially reduced over the other disulfide bonds in CD44 and reduction inhibits the CD44-HA interaction. Furthermore cells transfected with CD44 no longer adhere to HA coated surfaces after pre-treatment with reducing agents. The implications of CD44 redox regulation are discussed in the context of immune function, disease and therapeutic strategies. PMID:26379032

  15. The Effect of Hydrofluoric Acid Concentration on the Bond Strength and Morphology of the Surface and Interface of Glass Ceramics to a Resin Cement.

    PubMed

    Sundfeld Neto, D; Naves, L Z; Costa, A R; Correr, A B; Consani, S; Borges, G A; Correr-Sobrinho, L

    2015-01-01

    The purpose of this study was to evaluate the influence of various concentrations of hydrofluoric acid (HF) on the surface/interface morphology and μ-shear bond strength (μSBS) between IPS Empress Esthetic (EST) (Ivoclar Vivadent) and IPS e.max Press (EMX) (Ivoclar Vivadent) ceramics and resin cement. Ceramic blocks were divided into 12 groups for each kind of ceramic. Six different HF concentrations were evaluated: 1%, 2.5%, 5%, 7.5%, 10%, and 15%. All groups were silanated after etching, and half of the specimens within each group received a thin layer of unfilled resin (UR). Three resin cement cylinders were prepared on each ceramic block for μSBS testing. The specimens were stored in distilled water at 37°C for 24 hours. The μSBS test was carried out in a universal testing machine at a crosshead speed of 0.5 mm/min until fracture. The data were submitted to three-way analysis of variance and multiple comparisons were performed using the Tukey post hoc test (p<0.05). The etched surfaces and bonded interfaces were evaluated using scanning electron microscopy. μSBS means (MPa) for 1%, 2.5%, 5%, 7.5%, 10%, and 15% HF concentrations were, respectively, 25.2, 27.2, 30.1, 31.4, 33.3, and 31.8. μSBS means with or without UR application measured 32.24 and 27.4, respectively; EST and EMX measured 29.8 and 29.9, respectively. For the HF concentrations, 10% and 15% showed higher μSBS means than did 1% and 2.5% (p<0.05); 7.5% was higher than 1% (p<0.05); and no statistical differences were found among the other concentrations (p>0.05). When evaluating UR, μSBS mean was significantly higher and better infiltration was observed on the etched surfaces. No statistical difference was found between the ceramics. The HF concentration and UR influenced the bond strength and surface/interface morphology.

  16. Computational Identification of Protein Pupylation Sites by Using Profile-Based Composition of k-Spaced Amino Acid Pairs.

    PubMed

    Hasan, Md Mehedi; Zhou, Yuan; Lu, Xiaotian; Li, Jinyan; Song, Jiangning; Zhang, Ziding

    2015-01-01

    Prokaryotic proteins are regulated by pupylation, a type of post-translational modification that contributes to cellular function in bacterial organisms. In pupylation process, the prokaryotic ubiquitin-like protein (Pup) tagging is functionally analogous to ubiquitination in order to tag target proteins for proteasomal degradation. To date, several experimental methods have been developed to identify pupylated proteins and their pupylation sites, but these experimental methods are generally laborious and costly. Therefore, computational methods that can accurately predict potential pupylation sites based on protein sequence information are highly desirable. In this paper, a novel predictor termed as pbPUP has been developed for accurate prediction of pupylation sites. In particular, a sophisticated sequence encoding scheme [i.e. the profile-based composition of k-spaced amino acid pairs (pbCKSAAP)] is used to represent the sequence patterns and evolutionary information of the sequence fragments surrounding pupylation sites. Then, a Support Vector Machine (SVM) classifier is trained using the pbCKSAAP encoding scheme. The final pbPUP predictor achieves an AUC value of 0.849 in 10-fold cross-validation tests and outperforms other existing predictors on a comprehensive independent test dataset. The proposed method is anticipated to be a helpful computational resource for the prediction of pupylation sites. The web server and curated datasets in this study are freely available at http://protein.cau.edu.cn/pbPUP/.

  17. Counting Active Sites on Titanium Oxide-Silica Catalysts for Hydrogen Peroxide Activation through In Situ Poisoning with Phenylphosphonic Acid

    SciTech Connect

    Eaton, Todd R.; Boston, Andrew M.; Thompson, Anthony B.; Gray, Kimberly A.; Notestein, Justin M.

    2015-06-04

    Quantifying specific active sites in supported catalysts improves our understanding and assists in rational design. Supported oxides can undergo significant structural changes as surface densities increase from site-isolated cations to monolayers and crystallites, which changes the number of kinetically relevant sites. Herein, TiOx domains are titrated on TiOx–SiO2 selectively with phenylphosphonic acid (PPA). An ex situ method quantifies all fluid-accessible TiOx, whereas an in situ titration during cis-cyclooctene epoxidation provides previously unavailable values for the number of tetrahedral Ti sites on which H2O2 activation occurs. We use this method to determine the active site densities of 22 different catalysts with different synthesis methods, loadings, and characteristic spectra and find a single intrinsic turnover frequency for cis-cyclooctene epoxidation of (40±7) h-1. This simple method gives molecular-level insight into catalyst structure that is otherwise hidden when bulk techniques are used.

  18. Interaction of P-aminobenzoic acid with normal and sickel erythrocyte membrane: photoaffinity labelling of the binding sites

    SciTech Connect

    Premachandra, B.R.

    1986-03-05

    Electron microscopic studies revealed that P-Amino benzoic acid (PABA) could prevent eichinocytosis of red cells in vitro. Equilibrium binding studies with right side out membrane vesicles (ROV) revealed a similar number of binding sites (1.2-1.4 ..mu..mol/mg) and Kd (1.4-1.6 mM) values for both normal and sickle cell membranes. /sup 14/C-Azide analogue of PABA was synthesized as a photoaffinity label to probe its sites of interaction on the erythrocyte membranes. Competitive binding studies of PABA with its azide indicated that both the compounds share common binding sites on the membrane surface since a 20 fold excess of azide inhibited PABA binding in a linear fashion. The azide was covalently incorporated into the membrane components only upon irradiation (52-35% of the label found in the proteins and the rest in lipids). Electrophoretic analysis of photolabelled ROV revealed that the azide interacts chiefly with Band 3 protein. PABA inhibited both high and low affinity calcium (Ca) binding sites situated on either surface of the membrane in a non-competitive manner; however, Ca binding stimulated by Mg-ATP was not affected. Ca transport into inside out vesicles was inhibited by PABA; but it did not affect the calcium ATP-ase activity. The authors studies suggest that the mechanism of action of PABA is mediated by its interaction with Band 3 protein (anion channel), calcium channel and calcium binding sites of erythrocyte membrane.

  19. An active site mutant of Escherichia coli cyclopropane fatty acid synthase forms new non-natural fatty acids providing insights on the mechanism of the enzymatic reaction.

    PubMed

    E, Guangqi; Drujon, Thierry; Correia, Isabelle; Ploux, Olivier; Guianvarc'h, Dominique

    2013-12-01

    We have produced and purified an active site mutant of the Escherichia coli cyclopropane fatty acid synthase (CFAS) by replacing the strictly conserved G236 within cyclopropane synthases, by a glutamate residue, which corresponds to E146 of the homologous mycolic acid methyltransferase, Hma, producing hydroxymethyl mycolic acids. The G236E CFAS mutant had less than 1% of the in vitro activity of the wild type enzyme. We expressed the G236E CFAS mutant in an E. coli (DE3) strain in which the chromosomal cfa gene had been deleted. After extraction of phospholipids and conversion into the corresponding fatty acid methyl esters (FAMEs), we observed the formation of cyclopropanated FAMEs suggesting that the mutant retained some of the normal activity in vivo. However, we also observed the formation of new C17 methyl-branched unsaturated FAMEs whose structures were determined using GC/MS and NMR analyses. The double bond was located at different positions 8, 9 or 10, and the methyl group at position 10 or 9. Thus, this new FAMEs are likely arising from a 16:1 acyl chain of a phospholipid that had been transformed by the G236E CFAS mutant in vivo. The reaction catalyzed by this G236E CFAS mutant thus starts by the methylation of the unsaturated acyl chain at position 10 or 9 yielding a carbocation at position 9 or 10 respectively. It follows then two competing steps, a normal cyclopropanation or hydride shift/elimination events giving different combinations of alkenes. This study not only provides further evidence that cyclopropane synthases (CSs) form a carbocationic intermediate but also opens the way to CSs engineering for the synthesis of non-natural fatty acids.

  20. Does physical activity in adolescence have site-specific and sex-specific benefits on young adult bone size, content, and estimated strength?

    PubMed

    Duckham, Rachel L; Baxter-Jones, Adam D G; Johnston, James D; Vatanparast, Hassanali; Cooper, David; Kontulainen, Saija

    2014-02-01

    The long-term benefits of habitual physical activity during adolescence on adult bone structure and strength are poorly understood. We investigated whether physically active adolescents had greater bone size, density, content, and estimated bone strength in young adulthood when compared to their peers who were inactive during adolescence. Peripheral quantitative computed tomography (pQCT) was used to measure the tibia and radius of 122 (73 females) participants (age mean ± SD, 29.3 ± 2.3 years) of the Saskatchewan Pediatric Bone Mineral Accrual Study (PBMAS). Total bone area (ToA), cortical density (CoD), cortical area (CoA), cortical content (CoC), and estimated bone strength in torsion (SSIp ) and muscle area (MuA) were measured at the diaphyses (66% tibia and 65% radius). Total density (ToD), trabecular density (TrD), trabecular content (TrC), and estimated bone strength in compression (BSIc ) were measured at the distal ends (4%). Participants were grouped by their adolescent physical activity (PA) levels (inactive, average, and active) based on mean PA Z-scores obtained from serial questionnaire assessments completed during adolescence. We compared adult bone outcomes across adolescent PA groups in each sex using analysis of covariance followed by post hoc pairwise comparisons with Bonferroni adjustments. When adjusted for adult height, MuA, and PA, adult males who were more physically active than their peers in adolescence had 13% greater adjusted torsional bone strength (SSIp , p < 0.05) and 10% greater adjusted ToA (p < 0.05) at the tibia diaphysis. Females who were more active in adolescence had 10% larger adjusted CoA (p < 0.05), 12% greater adjusted CoC (p < 0.05) at the tibia diaphysis, and 3% greater adjusted TrC (p < 0.05) at the distal tibia when compared to their inactive peers. Benefits to tibia bone size, content, and strength in those who were more active during adolescence seemed to persist into young adulthood

  1. Strength nutrition.

    PubMed

    Volek, Jeff S

    2003-08-01

    Muscle strength is determined by muscle size and factors related to neural recruitment. Resistance training is a potent stimulus for increasing muscle size and strength. These increases are, to a large extent, influenced and mediated by changes in hormones that regulate important events during the recovery process following exercise. Provision of nutrients in the appropriate amounts and at the appropriate times is necessary to optimize the recovery process. This review discusses the results of research that has examined the potential for nutrition and dietary supplements to impact the acute response to resistance exercise and chronic adaptations to resistance training. To date, the most promising strategies to augment gains in muscle size and strength appear to be consumption of protein-carbohydrate calories before and after resistance exercise, and creatine supplementation.

  2. Analysis of a nucleotide-binding site of 5-lipoxygenase by affinity labelling: binding characteristics and amino acid sequences.

    PubMed Central

    Zhang, Y Y; Hammarberg, T; Radmark, O; Samuelsson, B; Ng, C F; Funk, C D; Loscalzo, J

    2000-01-01

    5-Lipoxygenase (5LO) catalyses the first two steps in the biosynthesis of leukotrienes, which are inflammatory mediators derived from arachidonic acid. 5LO activity is stimulated by ATP; however, a consensus ATP-binding site or nucleotide-binding site has not been found in its protein sequence. In the present study, affinity and photoaffinity labelling of 5LO with 5'-p-fluorosulphonylbenzoyladenosine (FSBA) and 2-azido-ATP showed that 5LO bound to the ATP analogues quantitatively and specifically and that the incorporation of either analogue inhibited ATP stimulation of 5LO activity. The stoichiometry of the labelling was 1.4 mol of FSBA/mol of 5LO (of which ATP competed with 1 mol/mol) or 0.94 mol of 2-azido-ATP/mol of 5LO (of which ATP competed with 0.77 mol/mol). Labelling with FSBA prevented further labelling with 2-azido-ATP, indicating that the same binding site was occupied by both analogues. Other nucleotides (ADP, AMP, GTP, CTP and UTP) also competed with 2-azido-ATP labelling, suggesting that the site was a general nucleotide-binding site rather than a strict ATP-binding site. Ca(2+), which also stimulates 5LO activity, had no effect on the labelling of the nucleotide-binding site. Digestion with trypsin and peptide sequencing showed that two fragments of 5LO were labelled by 2-azido-ATP. These fragments correspond to residues 73-83 (KYWLNDDWYLK, in single-letter amino acid code) and 193-209 (FMHMFQSSWNDFADFEK) in the 5LO sequence. Trp-75 and Trp-201 in these peptides were modified by the labelling, suggesting that they were immediately adjacent to the C-2 position of the adenine ring of ATP. Given the stoichiometry of the labelling, the two peptide sequences of 5LO were probably near each other in the enzyme's tertiary structure, composing or surrounding the ATP-binding site of 5LO. PMID:11042125

  3. Identification of Rosmarinic Acid-Adducted Sites in Meat Proteins in a Gel Model under Oxidative Stress by Triple TOF MS/MS.

    PubMed

    Tang, Chang-Bo; Zhang, Wan-Gang; Wang, Yao-Song; Xing, Lu-Juan; Xu, Xing-Lian; Zhou, Guang-Hong

    2016-08-24

    Triple TOF MS/MS was used to identify adducts between rosmarinic acid (RosA)-derived quinones and meat proteins in a gel model under oxidative stress. Seventy-five RosA-modified peptides responded to 67 proteins with adduction of RosA. RosA conjugated with different amino acids in proteins, and His, Arg, and Lys adducts with RosA were identified for the first time in meat. A total of 8 peptides containing Cys, 14 peptides containing His, 48 peptides containing Arg, 64 peptides containing Lys, and 5 peptides containing N-termini that which participated in adduction reaction with RosA were identified, respectively. Seventy-seven adduction sites were subdivided into all adducted proteins including 2 N-terminal adduction sites, 3 Cys adduction sites, 4 His adduction sites, 29 Arg adduction sites, and 39 Lys adduction sites. Site occupancy analyses showed that approximately 80.597% of the proteins carried a single RosA-modified site, 14.925% retained two sites, 1.492% contained three sites, and the rest 2.985% had four or more sites. Large-scale triple TOF MS/MS mapping of RosA-adducted sites reveals the adduction regulations of quinone and different amino acids as well as the adduction ratios, which clarify phenol-protein adductions and pave the way for industrial meat processing and preservation. PMID:27486909

  4. Evaluation of muscle strength and motor abilities in children with type II and III spinal muscle atrophy treated with valproic acid

    PubMed Central

    2011-01-01

    Background Spinal muscular atrophy (SMA) is an autosomal recessive disorder that affects the motoneurons of the spinal anterior horn, resulting in hypotonia and muscle weakness. The disease is caused by deletion or mutation in the telomeric copy of SMN gene (SMN1) and clinical severity is in part determined by the copy number of the centromeric copy of the SMN gene (SMN2). The SMN2 mRNA lacks exon 7, resulting in a production of lower amounts of the full-length SMN protein. Knowledge of the molecular mechanism of diseases has led to the discovery of drugs capable of increasing SMN protein level through activation of SMN2 gene. One of these drugs is the valproic acid (VPA), a histone deacetylase inhibitor. Methods Twenty-two patients with type II and III SMA, aged between 2 and 18 years, were treated with VPA and were evaluated five times during a one-year period using the Manual Muscle Test (Medical Research Council scale-MRC), the Hammersmith Functional Motor Scale (HFMS), and the Barthel Index. Results After 12 months of therapy, the patients did not gain muscle strength. The group of children with SMA type II presented a significant gain in HFMS scores during the treatment. This improvement was not observed in the group of type III patients. The analysis of the HFMS scores during the treatment period in the groups of patients younger and older than 6 years of age did not show any significant result. There was an improvement of the daily activities at the end of the VPA treatment period. Conclusion Treatment of SMA patients with VPA may be a potential alternative to alleviate the progression of the disease. Trial Registration ClinicalTrials.gov: NCT01033331 PMID:21435220

  5. Gas/particle partitioning of low-molecular-weight dicarboxylic acids at a suburban site in Saitama, Japan

    NASA Astrophysics Data System (ADS)

    Bao, Linfa; Matsumoto, Mariko; Kubota, Tsutomu; Sekiguchi, Kazuhiko; Wang, Qingyue; Sakamoto, Kazuhiko

    2012-02-01

    Low-molecular-weight dicarboxylic acids (diacids) exhibit semivolatile behavior in the atmosphere, but their partitioning between the gaseous and particulate phases is still unclear. An annular denuder-filter pack system with a cyclone PM 2.5 was employed to investigate the gaseous and particulate phase concentrations of diacids, with high collection efficiency of most target compounds. Saturated diacids, unsaturated diacids, ketocarboxylic acids, and dicarbonyls were determined in gaseous and particulate samples collected from a suburban site in Japan, during 2007 summer, 2008 late-winter and early-winter. The concentrations of gaseous and particulate diacids in early-winter were lower than those in summer, but higher than those in late-winter. Individual diacid in gaseous phase showed a relatively good correlation with ambient oxidants, but a low correlation with NO gas (a primary pollutant). Particulate fraction to the total amount ( FP) of individual acid was larger in winter than in summer, and also was larger at night than in the daytime. In the same sample, individual diacid and ketocarboxylic acid had higher particulate phase occurrence ( FP > 56% in summer), whereas unsaturated diacid had higher gaseous phase occurrence ( FP < 18% in summer). In summer, gas/particle partitioning of diacids varied diurnally; FP values of oxalic and glyoxylic acids increased from their lowest values in the morning to their highest values at night, exhibiting the similar diurnal variation of relative humidity in the atmosphere. The higher humidity at night may lead to the formation of droplets in which water-soluble gaseous phases can dissolve, thus promoting gas-to-particle conversion. These results suggest that gas/particle partitioning of diacids depends not only on the concentrations in the gaseous phase by photochemical oxidation, but also on the characteristics of the atmosphere (e.g., temperature, sunlight, and relative humidity) and the aerosols (e.g., acidity

  6. Boron and calcium sites involved in indole-3-acetic Acid transport in sunflower hypocotyl segments.

    PubMed

    Tang, P M; Dela Fuente, R K

    1986-06-01

    Sunflower (Helianthus annuus L. cv Russian Mammoth) hypocotyl segments deficient in either B or Ca exhibited a higher rate of potassium leakage, compared to nondeficient segments. Potassium leakage, used here as an indication of membrane integrity, was completely reversed by the addition of H(3)BO(3) or Ca(NO(3))(2) to the incubation medium of the B-deficient or Ca-deficient hypocotyl segments, respectively. This role of B and Ca in membrane integrity, which may be important in the entry and exit of auxin in cells, is identified as the first site of action for each of these two essential elements in the basipetal secretion of auxin. A second site for B is postulated because auxin transport was not restored, even when K(+) leakage has been completely reversed to the nondeficient level, when B-deficient hypocotyls were incubated in B solution. This lack of reversibility of auxin transport implied that the incubation for 2 h in B solution was not enough to restore the auxin transport process. However, since the transfer of B-deficient seedlings to B solutions prevented further deterioration of auxin transport, these observations suggest that: (a) either an intact seedling, or a longer period of incubation of the hypocotyl in B solution, is required for the synthesis or maintenance of the functional second site for B; (b) B is probably essential in the synthesis of a ligand, which may or may not be needed to bind B, but which is essential in the basipetal transport of auxin. The second site for Ca in auxin transport, is indicated by the complete reversal of its inhibition in Ca-deficient hypocotyl, when incubated in Ca solution. The second site for Ca is thought to be directly involved in the secretion of auxin, in which Ca probably plays the role of a second messenger, as in stimulus-response coupling. The two sites for Ca can be distinguished from each other by their cation specificity. The requirement for Ca in the first site can be substituted by other divalent

  7. Histomorphological researches on large porous hydroxyapatite cylinder tubes with polylactic acid surface coating in different nonskeletal sites in vivo.

    PubMed

    Zhang, Cong; Huang, Peng; Weng, Jie; Zhi, Wei; Hu, Yonghe; Feng, Huaizhi; Yao, Yimin; Li, Shuo; Xia, Tian

    2012-05-01

    Porous hydroxyapatite (HA) ceramic cylinder tubes coated with polylactic acid on the exposed surfaces were implanted in four nonskeletal sites (omentum, peritoneum, vastus lateralis, and side of femur). Six months postoperatively, proper amount of Chinese ink was injected to dye the implanting areas. Decalcified and nondecalcified sections were observed under inverted microscope. The results showed that the soft tissues around the HA cylinder tubes in peritoneum, vastus lateralis, and side of femur groups appeared visible black. Some small blacked vascular architectures were also discernible. However in omentum group, only small number of blacked vessels existed. Histological observations indicated that vascularization and ossification occurred in peritoneum, vastus lateralis, and side of femur groups. In omentum group, there was no any sign of vascularization and ossification. A conclusion could be made in this study that excepting bones and muscles, parietal peritoneum could serve as a potential spot for culturing histoengineering hydroxyapatite (HA)-polylactic acid (PLA) ceramic bone substitutes.

  8. Histomorphological researches on large porous hydroxyapatite cylinder tubes with polylactic acid surface coating in different nonskeletal sites in vivo.

    PubMed

    Zhang, Cong; Huang, Peng; Weng, Jie; Zhi, Wei; Hu, Yonghe; Feng, Huaizhi; Yao, Yimin; Li, Shuo; Xia, Tian

    2012-05-01

    Porous hydroxyapatite (HA) ceramic cylinder tubes coated with polylactic acid on the exposed surfaces were implanted in four nonskeletal sites (omentum, peritoneum, vastus lateralis, and side of femur). Six months postoperatively, proper amount of Chinese ink was injected to dye the implanting areas. Decalcified and nondecalcified sections were observed under inverted microscope. The results showed that the soft tissues around the HA cylinder tubes in peritoneum, vastus lateralis, and side of femur groups appeared visible black. Some small blacked vascular architectures were also discernible. However in omentum group, only small number of blacked vessels existed. Histological observations indicated that vascularization and ossification occurred in peritoneum, vastus lateralis, and side of femur groups. In omentum group, there was no any sign of vascularization and ossification. A conclusion could be made in this study that excepting bones and muscles, parietal peritoneum could serve as a potential spot for culturing histoengineering hydroxyapatite (HA)-polylactic acid (PLA) ceramic bone substitutes. PMID:22344718

  9. Hydrogenation of biofuels with formic acid over a palladium-based ternary catalyst with two types of active sites.

    PubMed

    Wang, Liang; Zhang, Bingsen; Meng, Xiangju; Su, Dang Sheng; Xiao, Feng-Shou

    2014-06-01

    A composite catalyst including palladium nanoparticles on titania (TiO2) and on nitrogen-modified porous carbon (Pd/TiO2@N-C) is synthesized from palladium salts, tetrabutyl titanate, and chitosan. N2 sorption isotherms show that the catalyst has a high BET surface area (229 m(2)  g(-1)) and large porosity. XPS and TEM characterization of the catalyst shows that palladium species with different chemical states are well dispersed across the TiO2 and nitrogen-modified porous carbon, respectively. The Pd/TiO2@N-C catalyst is very active and shows excellent stability towards hydrogenation of vanillin to 2-methoxy-4-methylphenol using formic acid as hydrogen source. This activity can be attributed to a synergistic effect between the Pd/TiO2 (a catalyst for dehydrogenation of formic acid) and Pd/N-C (a catalyst for hydrogenation of vanillin) sites.

  10. Chicoric acid binds to two sites and decreases the activity of the YopH bacterial virulence factor

    PubMed Central

    Kuban-Jankowska, Alicja; Sahu, Kamlesh K.; Gorska, Magdalena; Tuszynski, Jack A.; Wozniak, Michal

    2016-01-01

    Chicoric acid (CA) is a phenolic compound present in dietary supplements with a large spectrum of biological properties reported ranging from antioxidant, to antiviral, to immunostimulatory properties. Due to the fact that chicoric acid promotes phagocytic activity and was reported as an allosteric inhibitor of the PTP1B phosphatase, we examined the effect of CA on YopH phosphatase from pathogenic bacteria, which block phagocytic processes of a host cell. We performed computational studies of chicoric acid binding to YopH as well as validation experiments with recombinant enzymes. In addition, we performed similar studies for caffeic and chlorogenic acids to compare the results. Docking experiments demonstrated that, from the tested compounds, only CA binds to both catalytic and secondary binding sites of YopH. Our experimental results showed that CA reduces activity of recombinant YopH phosphatase from Yersinia enterocolitica and human CD45 phosphatase. The inhibition caused by CA was irreversible and did not induce oxidation of catalytic cysteine. We proposed that inactivation of YopH induced by CA is involved with allosteric inhibition by interacting with essential regions responsible for ligand binding. PMID:26735581

  11. Experimental and Mechanistic Understanding of Aldehyde Hydrogenation Using Au25 Nanoclusters with Lewis Acids: Unique Sites for Catalytic Reactions.

    PubMed

    Li, Gao; Abroshan, Hadi; Chen, Yuxiang; Jin, Rongchao; Kim, Hyung J

    2015-11-18

    The catalytic activity of Au25(SR)18 nanoclusters (R = C2H4Ph) for the aldehyde hydrogenation reaction in the presence of a base, e.g., ammonia or pyridine, and transition-metal ions M(z+), such as Cu(+), Cu(2+), Ni(2+) and Co(2+), as a Lewis acid is studied. The addition of a Lewis acid is found to significantly promote the catalytic activity of Au25(SR)18/CeO2 in the hydrogenation of benzaldehyde and a number of its derivatives. Matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) mass spectrometry in conjunction with UV-vis spectroscopy confirm the generation of new species, Au25-n(SR)18-n (n = 1-4), in the presence of a Lewis acid. The pathways for the speciation of Au24(SR)17 from its parent Au25(SR)18 nanocluster as well as its structure are investigated via the density functional theory (DFT) method. The adsorption of M(z+) onto a thiolate ligand "-SR-" of Au25(SR)18, followed by a stepwise detachment of "-SR-" and a gold atom bonded to "-SR-" (thus an "Au-SR" unit) is found to be the most likely mechanism for the Au24(SR)17 generation. This in turn exposes the Au13-core of Au24(SR)17 to reactants, providing an active site for the catalytic hydrogenation. DFT calculations indicate that M(z+) is also capable of adsorbing onto the Au13-core surface, producing a possible active metal site of a different kind to catalyze the aldehyde hydrogenation reaction. This study suggests, for the first time, that species with an open metal site like adducts [nanoparticle-M]((z-1)+) or fragments Au25-n(SR)18-n function as the catalysts rather than the intact Au25(SR)18.

  12. Remediation of acid mine drainage at the friendship hill national historic site with a pulsed limestone bed process

    USGS Publications Warehouse

    Sibrell, P.L.; Watten, B.; Boone, T.; ,

    2003-01-01

    A new process utilizing pulsed fluidized limestone beds was tested for the remediation of acid mine drainage at the Friendship Hill National Historic Site, in southwestern Pennsylvania. A 230 liter-per-minute treatment system was constructed and operated over a fourteen-month period from June 2000 through September 2001. Over this period of time, 50,000 metric tons of limestone were used to treat 50 million liters of water. The influent water pH was 2.5 and acidity was 1000 mg/L as CaCO3. Despite the high potential for armoring at the site, effluent pH during normal plant operation ranged from 5.7 to 7.8 and averaged 6.8. As a result of the high influent acidity, sufficient CO2 was generated and recycled to provide a net alkaline discharge with about 50 mg/L as CaCO3 alkalinity. Additions of commercial CO2 increased effluent alkalinity to as high as 300 mg/L, and could be a useful process management tool for transient high flows or acidities. Metal removal rates were 95% for aluminum (60 mg/L in influent), 50 to 90% for iron (Fe), depending on the ratio of ferrous to ferric iron, which varied seasonally (200 mg/L in influent), and <10% of manganese (Mn) (10 mg/L in influent). Ferrous iron and Mn removal was incomplete because of the high pH required for precipitation of these species. Iron removal could be improved by increased aeration following neutralization, and Mn removal could be effected by a post treatment passive settling/oxidation pond. Metal hydroxide sludges were settled in settling tanks, and then hauled from the site for aesthetic purposes. Over 450 metric tons of sludge were removed from the water over the life of the project. The dried sludge was tested by the Toxicity Characteristics Leaching Protocol (TCLP) and was found to be non-hazardous. Treatment costs were $43,000 per year and $1.08 per m 3, but could be decreased to $22,000 and $0.51 per m3 by decreasing labor use and by onsite sludge handling. These results confirm the utility of the new

  13. Technology development for phosphoric acid fuel cell powerplant (phase 2). [on site integrated energy systems

    NASA Technical Reports Server (NTRS)

    Christner, L.

    1980-01-01

    Progress is reported in the development of material, cell components, and reformers for on site integrated energy systems. Internal resistance and contact resistance were improved. Dissolved gases (O2, N2, and CO2) were found to have no effect on the electrochemical corrosion of phenolic composites. Stack performance was increased by 100 mV over the average 1979 level.

  14. Site-Specifically Labeled Immunoconjugates for Molecular Imaging--Part 2: Peptide Tags and Unnatural Amino Acids.

    PubMed

    Adumeau, Pierre; Sharma, Sai Kiran; Brent, Colleen; Zeglis, Brian M

    2016-04-01

    Molecular imaging using radioisotope- or fluorophore-labeled antibodies is increasingly becoming a critical component of modern precision medicine. Yet despite this promise, the vast majority of these immunoconjugates are synthesized via the random coupling of amine-reactive bifunctional probes to lysines within the antibody, a process that can result in heterogeneous and poorly defined constructs with suboptimal pharmacological properties. In an effort to circumvent these issues, the last 5 years have played witness to a great deal of research focused on the creation of effective strategies for the site-specific attachment of payloads to antibodies. These chemoselective modification methods yield immunoconjugates that are more homogenous and better defined than constructs created using traditional synthetic approaches. Moreover, site-specifically labeled immunoconjugates have also been shown to exhibit superior in vivo behavior compared to their randomly modified cousins. The over-arching goal of this two-part review is to provide a broad yet detailed account of the various site-specific bioconjugation approaches that have been used to create immunoconjugates for positron emission tomography (PET), single photon emission computed tomography (SPECT), and fluorescence imaging. In Part 1, we covered site-specific bioconjugation techniques based on the modification of cysteine residues and the chemoenzymatic manipulation of glycans. In Part 2, we will detail two families of bioconjugation approaches that leverage biochemical tools to achieve site-specificity. First, we will discuss modification methods that employ peptide tags either as sites for enzyme-catalyzed ligations or as radiometal coordination architectures. And second, we will examine bioconjugation strategies predicated on the incorporation of unnatural or non-canonical amino acids into antibodies via genetic engineering. Finally, we will compare the advantages and disadvantages of the modification

  15. Site-Specifically Labeled Immunoconjugates for Molecular Imaging—Part 2: Peptide Tags and Unnatural Amino Acids

    PubMed Central

    Adumeau, Pierre; Sharma, Sai Kiran; Brent, Colleen; Zeglis, Brian M.

    2016-01-01

    Molecular imaging using radioisotope- or fluorophore-labeled antibodies is increasingly becoming a critical component of modern precision medicine. Yet despite this promise, the vast majority of these immunoconjugates are synthesized via the random coupling of amine-reactive bifunctional probes to lysines within the antibody, a process that can result in heterogeneous and poorly defined constructs with suboptimal pharmacological properties. In an effort to circumvent these issues, the last 5 years have played witness to a great deal of research focused on the creation of effective strategies for the site-specific attachment of payloads to antibodies. These chemoselective modification methods yield immunoconjugates that are more homogenous and better defined than constructs created using traditional synthetic approaches. Moreover, site-specifically labeled immunoconjugates have also been shown to exhibit superior in vivo behavior compared to their randomly modified cousins. The over-arching goal of this two-part review is to provide a broad yet detailed account of the various site-specific bioconjugation approaches that have been used to create immunoconjugates for positron emission tomography (PET), single photon emission computed tomography (SPECT), and fluorescence imaging. In Part 1, we covered site-specific bioconjugation techniques based on the modification of cysteine residues and the chemoenzymatic manipulation of glycans. In Part 2, we will detail two families of bioconjugation approaches that leverage biochemical tools to achieve site-specificity. First, we will discuss modification methods that employ peptide tags either as sites for enzyme-catalyzed ligations or as radiometal coordination architectures. And second, we will examine bioconjugation strategies predicated on the incorporation of unnatural or non-canonical amino acids into antibodies via genetic engineering. Finally, we will compare the advantages and disadvantages of the modification

  16. The Significance of Lewis Acid Sites for the Selective Catalytic Reduction of Nitric Oxide on Vanadium-Based Catalysts.

    PubMed

    Marberger, Adrian; Ferri, Davide; Elsener, Martin; Kröcher, Oliver

    2016-09-19

    The long debated reaction mechanisms of the selective catalytic reduction (SCR) of nitric oxide with ammonia (NH3 ) on vanadium-based catalysts rely on the involvement of Brønsted or Lewis acid sites. This issue has been clearly elucidated using a combination of transient perturbations of the catalyst environment with operando time-resolved spectroscopy to obtain unique molecular level insights. Nitric oxide reacts predominantly with NH3 coordinated to Lewis sites on vanadia on tungsta-titania (V2 O5 -WO3 -TiO2 ), while Brønsted sites are not involved in the catalytic cycle. The Lewis site is a mono-oxo vanadyl group that reduces only in the presence of both nitric oxide and NH3 . We were also able to verify the formation of the nitrosamide (NH2 NO) intermediate, which forms in tandem with vanadium reduction, and thus the entire mechanism of SCR. Our experimental approach, demonstrated in the specific case of SCR, promises to progress the understanding of chemical reactions of technological relevance.

  17. Structure and Mutagenesis of Neural Cell Adhesion Molecule Domains Evidence for Flexibility in the Placement of Polysialic Acid Attachment Sites

    SciTech Connect

    Foley, Deirdre A.; Swartzentruber, Kristin G.; Lavie, Arnon; Colley, Karen J.

    2010-11-09

    The addition of {alpha}2,8-polysialic acid to the N-glycans of the neural cell adhesion molecule, NCAM, is critical for brain development and plays roles in synaptic plasticity, learning and memory, neuronal regeneration, and the growth and invasiveness of cancer cells. Our previous work indicates that the polysialylation of two N-glycans located on the fifth immunoglobulin domain (Ig5) of NCAM requires the presence of specific sequences in the adjacent fibronectin type III repeat (FN1). To understand the relationship of these two domains, we have solved the crystal structure of the NCAM Ig5-FN1 tandem. Unexpectedly, the structure reveals that the sites of Ig5 polysialylation are on the opposite face from the FN1 residues previously found to be critical for N-glycan polysialylation, suggesting that the Ig5-FN1 domain relationship may be flexible and/or that there is flexibility in the placement of Ig5 glycosylation sites for polysialylation. To test the latter possibility, new Ig5 glycosylation sites were engineered and their polysialylation tested. We observed some flexibility in glycosylation site location for polysialylation and demonstrate that the lack of polysialylation of a glycan attached to Asn-423 may be in part related to a lack of terminal processing. The data also suggest that, although the polysialyltransferases do not require the Ig5 domain for NCAM recognition, their ability to engage with this domain is necessary for polysialylation to occur on Ig5 N-glycans.

  18. The Significance of Lewis Acid Sites for the Selective Catalytic Reduction of Nitric Oxide on Vanadium-Based Catalysts.

    PubMed

    Marberger, Adrian; Ferri, Davide; Elsener, Martin; Kröcher, Oliver

    2016-09-19

    The long debated reaction mechanisms of the selective catalytic reduction (SCR) of nitric oxide with ammonia (NH3 ) on vanadium-based catalysts rely on the involvement of Brønsted or Lewis acid sites. This issue has been clearly elucidated using a combination of transient perturbations of the catalyst environment with operando time-resolved spectroscopy to obtain unique molecular level insights. Nitric oxide reacts predominantly with NH3 coordinated to Lewis sites on vanadia on tungsta-titania (V2 O5 -WO3 -TiO2 ), while Brønsted sites are not involved in the catalytic cycle. The Lewis site is a mono-oxo vanadyl group that reduces only in the presence of both nitric oxide and NH3 . We were also able to verify the formation of the nitrosamide (NH2 NO) intermediate, which forms in tandem with vanadium reduction, and thus the entire mechanism of SCR. Our experimental approach, demonstrated in the specific case of SCR, promises to progress the understanding of chemical reactions of technological relevance. PMID:27553251

  19. Capture and Recycling of Sortase A through Site-Specific Labeling with Lithocholic Acid.

    PubMed

    Rosen, Christian B; Kwant, Richard L; MacDonald, James I; Rao, Meera; Francis, Matthew B

    2016-07-18

    Enzyme-mediated protein modification often requires large amounts of biocatalyst, adding significant costs to the process and limiting industrial applications. Herein, we demonstrate a scalable and straightforward strategy for the efficient capture and recycling of enzymes using a small-molecule affinity tag. A proline variant of an evolved sortase A (SrtA 7M) was N-terminally labeled with lithocholic acid (LA)-an inexpensive bile acid that exhibits strong binding to β-cyclodextrin (βCD). Capture and recycling of the LA-Pro-SrtA 7M conjugate was achieved using βCD-modified sepharose resin. The LA-Pro-SrtA 7M conjugate retained full enzymatic activity, even after multiple rounds of recycling. PMID:27239057

  20. Fixed-site ion exchanger for liquid chromatographic determination of multifunctional carboxylic acids

    SciTech Connect

    Cassidy, R.M.; Elchuk, S.

    1985-03-01

    Reversed phases coated with a permanently sorbed ion exchanger and indirect UV detection have been investigated for the determination of simple and multifunctional carboxylic acids in chemical cleaning solutions. The advantages of being able to vary both the ion-exchange capacity and the hydrophobic interactions on these types of ion exchangers for the optimization of resolution and detection are illustrated, and the selection of optimum separation conditions is discussed. Dissolved iron interferes with the analysis due to photochemical, redox, and kinetic effects but good recoveries can be obtained after reduction of the iron with hydroxylamine and complexation with 1,2-diaminocyclohexanetetraacetic acid. Detection limits (3 x base line noise) for oxalate, citrate, ethylenediaminetetraacetate, and hydroxyethylenediaminetriacetate are 0.6-20 ..mu..g x mL/sup -1/ for a 20-..mu..L sample, and relative standard deviations are 3 to % in the 75-350 ..mu..g x mL/sup -1/ range. Analysis results for reactor decontamination solutions containing up to 250 ..mu..g x mL/sup -1/ of iron agree with results obtained by other techniques, and it is shown that this technique should also be useful for determination of metal ions in the samples. A determination of the above reagents in the presence of Fe(II) and Ni(II) takes 7 to 12 min after a 5 to 10 min reduction step. Cr(III) forms nonlabile complexes with ethylenediaminetetraacetic acid, and its presence will cause low results for this acid. 17 references, 4 figures, 6 tables.

  1. One site is enough: a theoretical investigation of iron-catalyzed dehydrogenation of formic Acid.

    PubMed

    Sánchez-de-Armas, Rocío; Xue, Liqin; Ahlquist, Mårten S G

    2013-09-01

    Dehydrogenation of HCO2H: The reaction mechanism for the dehydrogenation of formic acid catalyzed by a highly active and selective iron complex has been studied by DFT. The most favorable pathway shows the hydride in Fe-H complexes acting as a spectator ligand throughout the catalytic cycle. This result opens up the Fe complex for modification in order to achieve more efficient and selective catalysts. PMID:23907850

  2. Metal ion blockage of tritium incorporation into gamma-carboxyglutamic acid of prothrombin. Stoichiometry of gamma-carboxyglutamic acid to Gd3+ for the high affinity sites

    SciTech Connect

    Bajaj, S.P.; Saini, R.; Katz, A.; Cai, G.Z.; Maki, S.L.; Brodsky, G.L.

    1988-07-15

    Prothrombin possesses two high affinity and four low affinity gamma-carboxyglutamic acid (Gla)-dependent gadolinium binding sites. Earlier work has shown that tritium can be specifically incorporated at the gamma-carbon of Gla in proteins at pH 5. In the present work we show that inclusion of saturating concentrations of Ca2+ in nondenaturing buffer systems ranging from pH 5.5 to 8.5 prevents the exchange of tritium into all 10 Gla residues of prothrombin. Similarly, saturating concentrations of Gd3+ prevent tritium incorporation into Gla at pH 5.5. Positive cooperativity was observed for the binding of Gd3+ to human prothrombin (at pH 5.5) for the two high affinity sites (Kd congruent to 35 nM). The four low affinity sites bind Gd3+ with a Kd congruent to 5 microM. Incubation of prothrombin ranging in concentrations from 10 to 40 microM with 2 eq of Gd3+ at pH 5.5 prevents 5.7 (average of seven determinations) Gla residues from tritium incorporation. Sedimentation velocity experiments conducted at pH 5.5 indicate that prothrombin in the presence of saturating concentrations of Gd3+ polymerizes, most likely, to a trimer. Further, in the presence of 2 eq of Gd3+, calculated percent weight average concentration of monomer prothrombin is congruent to 100% at 10 microM, approximately equal to 95% at 20 microM, and congruento to 80% at 40 microM protein concentration. Thus, it appears that under conditions in which prothrombin primarily exists as a monomer, occupancy of the initial two metal binding sites by Gd3+ involves six Gla residues.

  3. Heterogeneous Chemistry of Lipopolysaccharides with Gas-Phase Nitric Acid: Reactive Sites and Reaction Pathways.

    PubMed

    Trueblood, Jonathan V; Estillore, Armando D; Lee, Christopher; Dowling, Jacqueline A; Prather, Kimberly A; Grassian, Vicki H

    2016-08-18

    Recent studies have shown that sea spray aerosol (SSA) has a size-dependent, complex composition consisting of biomolecules and biologically derived organic compounds in addition to salts. This additional chemical complexity most likely influences the heterogeneous reactivity of SSA, as these other components will have different reactive sites and reaction pathways. In this study, we focus on the reactivity of a class of particles derived from some of the biological components of sea spray aerosol including lipopolysaccharides (LPS) that undergo heterogeneous chemistry within the reactive sites of the biological molecule. Examples of these reactions and the relevant reactive sites are proposed as follows: R-COONa(s) + HNO3(g) → NaNO3 + R-COOH and R-HPO4Na(s) + HNO3(g) → NaNO3 + R-H2PO4. These reactions may be a heterogeneous pathway not only for sea spray aerosol but also for a variety of other types of atmospheric aerosol as well.

  4. The Association between Elevated Levels of Peripheral Serotonin and Its Metabolite – 5-Hydroxyindoleacetic Acid and Bone Strength and Metabolism in Growing Rats with Mild Experimental Chronic Kidney Disease

    PubMed Central

    Oksztulska-Kolanek, Ewa; Znorko, Beata; Domaniewski, Tomasz; Rogalska, Joanna; Roszczenko, Alicja; Brzóska, Małgorzata Michalina; Pryczynicz, Anna; Kemona, Andrzej

    2016-01-01

    Chronic kidney disease (CKD) is associated with disturbances in bone strength and metabolism. The alterations of the serotonergic system are also observed in CKD. We used the 5/6 nephrectomy model of CKD to assess the impact of peripheral serotonin and its metabolite– 5-hydroxyindoleacetic acid on bone biomechanical properties and metabolism in growing rats. The animals were sacrificed one and three months after nephrectomy. Biomechanical properties were determined on two different bone types: the cortical bone of the femoral diaphysis using three-point bending test and the mixed cortico-trabecular bone by the bending test of the femoral neck. Biomechanical tests revealed preserved cortical bone strength, whereas work to fracture (W) and yield load (Fy) of mixed cortico-trabecular bone were significantly lower in CKD compared to controls. Serum activity of alkaline phosphatase (ALP), a bone formation marker, and tartrate-resistant acid phosphatase (TRACP 5b) reflecting bone resorption, were similar in CKD and controls. ALP was associated with lower femoral stiffness and strength, and higher displacements and W. TRACP 5b was inversely associated with cortical Fu and W. The elevated peripheral serotonergic system in CKD was: inversely associated with stiffness but positively related to the displacements and W; inversely associated with cortical Fy but positively correlated with this parameter in cortico-trabecular bone; inversely associated with ALP in controls but positively correlated with this biomarker in CKD animals. In conclusion, this study demonstrates the distinct effect of mild degree of CKD on bone strength in rapidly growing rats. The impaired renal function affects the peripheral serotonin metabolism, which in turn may influence the strength and metabolism of bones in these rats. This relationship seems to be beneficial on the biomechanical properties of the cortico-trabecular bone, whereas the cortical bone strength can be potentially reduced. PMID

  5. Distinguishing two groups of flavin reductases by analyzing the protonation state of an active site carboxylic acid.

    PubMed

    Dumit, Verónica I; Cortez, Néstor; Matthias Ullmann, G

    2011-07-01

    Flavin-containing reductases are involved in a wide variety of physiological reactions such as photosynthesis, nitric oxide synthesis, and detoxification of foreign compounds, including therapeutic drugs. Ferredoxin-NADP(H)-reductase (FNR) is the prototypical enzyme of this family. The fold of this protein is highly conserved and occurs as one domain of several multidomain enzymes such as the members of the diflavin reductase family. The enzymes of this family have emerged as fusion of a FNR and a flavodoxin. Although the active sites of these enzymes are very similar, different enzymes function in opposite directions, that is, some reduce oxidized nicotinamide adenine dinucleotide phosphate (NADP(+)) and some oxidize reduced nicotinamide adenine dinucleotide phosphate (NADPH). In this work, we analyze the protonation behavior of titratable residues of these enzymes through electrostatic calculations. We find that a highly conserved carboxylic acid in the active site shows a different titration behavior in different flavin reductases. This residue is deprotonated in flavin reductases present in plastids, but protonated in bacterial counterparts and in diflavin reductases. The protonation state of the carboxylic acid may also influence substrate binding. The physiological substrate for plastidic enzymes is NADP(+), but it is NADPH for the other mentioned reductases. In this article, we discuss the relevance of the environment of this residue for its protonation and its importance in catalysis. Our results allow to reinterpret and explain experimental data. PMID:21538544

  6. Impact of acid mine drainages on surficial waters of an abandoned mining site.

    PubMed

    García-Lorenzo, M L; Marimón, J; Navarro-Hervás, M C; Pérez-Sirvent, C; Martínez-Sánchez, M J; Molina-Ruiz, José

    2016-04-01

    Weathering of sulphide minerals produces a great variety of efflorescences of soluble sulphate salts. These minerals play an important role for environmental pollution, since they can be either a sink or a source for acidity and trace elements. This paper aims to characterise surface waters affected by mining activities in the Sierra Minera of Cartagena-La Union (SE, Spain). Water samples were analysed for trace metals (Zn, Cd, Pb, Cu, As and Fe), major ions (Na(+), K(+), Ca(2+) and Mg(2+)) and anions (F(-), Cl(-), NO3 (-), CO3 (2-), SO4 (2-)) concentrations and were submitted to an "evaporation-precipitation" experiment that consisted in identifying the salts resulting from the evaporation of the water aliquots sampled onsite. Mineralogy of the salts was studied using X-ray diffraction and compared with the results of calculations using VISUAL MINTEQ. The study area is heavily polluted as a result of historical mining and processing activities that has produced large amount of wastes characterised by a high trace elements content, acidic pH and containing minerals resulting from the supergene alteration of the raw materials. The mineralogical study of the efflorescences obtained from waters shows that magnesium, zinc, iron and aluminium sulphates predominate in the acid mine drainage precipitates. Minerals of the hexahydrite group have been quantified together with minerals of the rozenite group, alunogen and other phases such as coquimbite and copiapite. Calcium sulphates correspond exclusively to gypsum. In a semiarid climate, such as that of the study area, these minerals contribute to understand the response of the system to episodic rainfall events. MINTEQ model could be used for the analysis of waters affected by mining activities but simulation of evaporation gives more realistic results considering that MINTEQ does not consider soluble hydrated salts.

  7. Impact of acid mine drainages on surficial waters of an abandoned mining site.

    PubMed

    García-Lorenzo, M L; Marimón, J; Navarro-Hervás, M C; Pérez-Sirvent, C; Martínez-Sánchez, M J; Molina-Ruiz, José

    2016-04-01

    Weathering of sulphide minerals produces a great variety of efflorescences of soluble sulphate salts. These minerals play an important role for environmental pollution, since they can be either a sink or a source for acidity and trace elements. This paper aims to characterise surface waters affected by mining activities in the Sierra Minera of Cartagena-La Union (SE, Spain). Water samples were analysed for trace metals (Zn, Cd, Pb, Cu, As and Fe), major ions (Na(+), K(+), Ca(2+) and Mg(2+)) and anions (F(-), Cl(-), NO3 (-), CO3 (2-), SO4 (2-)) concentrations and were submitted to an "evaporation-precipitation" experiment that consisted in identifying the salts resulting from the evaporation of the water aliquots sampled onsite. Mineralogy of the salts was studied using X-ray diffraction and compared with the results of calculations using VISUAL MINTEQ. The study area is heavily polluted as a result of historical mining and processing activities that has produced large amount of wastes characterised by a high trace elements content, acidic pH and containing minerals resulting from the supergene alteration of the raw materials. The mineralogical study of the efflorescences obtained from waters shows that magnesium, zinc, iron and aluminium sulphates predominate in the acid mine drainage precipitates. Minerals of the hexahydrite group have been quantified together with minerals of the rozenite group, alunogen and other phases such as coquimbite and copiapite. Calcium sulphates correspond exclusively to gypsum. In a semiarid climate, such as that of the study area, these minerals contribute to understand the response of the system to episodic rainfall events. MINTEQ model could be used for the analysis of waters affected by mining activities but simulation of evaporation gives more realistic results considering that MINTEQ does not consider soluble hydrated salts. PMID:26347422

  8. Incorporating significant amino acid pairs to identify O-linked glycosylation sites on transmembrane proteins and non-transmembrane proteins

    PubMed Central

    2010-01-01

    Background While occurring enzymatically in biological systems, O-linked glycosylation affects protein folding, localization and trafficking, protein solubility, antigenicity, biological activity, as well as cell-cell interactions on membrane proteins. Catalytic enzymes involve glycotransferases, sugar-transferring enzymes and glycosidases which trim specific monosaccharides from precursors to form intermediate structures. Due to the difficulty of experimental identification, several works have used computational methods to identify glycosylation sites. Results By investigating glycosylated sites that contain various motifs between Transmembrane (TM) and non-Transmembrane (non-TM) proteins, this work presents a novel method, GlycoRBF, that implements radial basis function (RBF) networks with significant amino acid pairs (SAAPs) for identifying O-linked glycosylated serine and threonine on TM proteins and non-TM proteins. Additionally, a membrane topology is considered for reducing the false positives on glycosylated TM proteins. Based on an evaluation using five-fold cross-validation, the consideration of a membrane topology can reduce 31.4% of the false positives when identifying O-linked glycosylation sites on TM proteins. Via an independent test, GlycoRBF outperforms previous O-linked glycosylation site prediction schemes. Conclusion A case study of Cyclic AMP-dependent transcription factor ATF-6 alpha was presented to demonstrate the effectiveness of GlycoRBF. Web-based GlycoRBF, which can be accessed at http://GlycoRBF.bioinfo.tw, can identify O-linked glycosylated serine and threonine effectively and efficiently. Moreover, the structural topology of Transmembrane (TM) proteins with glycosylation sites is provided to users. The stand-alone version of GlycoRBF is also available for high throughput data analysis. PMID:21034461

  9. Genetic Incorporation of the Unnatural Amino Acid p-Acetyl Phenylalanine into Proteins for Site-Directed Spin Labeling

    PubMed Central

    Evans, Eric G.B.; Millhauser, Glenn L.

    2016-01-01

    Site-directed spin labeling (SDSL) is a powerful tool for the characterization of protein structure and dynamics; however, its application in many systems is hampered by the reliance on unique and benign cysteine substitutions for the site-specific attachment of the spin label. An elegant solution to this problem involves the use of genetically encoded unnatural amino acids (UAAs) containing reactive functional groups that are chemically orthogonal to those of the 20 amino acids found naturally in proteins. These unique functional groups can then be selectively reacted with an appropriately functionalized spin probe. In this chapter, we detail the genetic incorporation of the ketone-bearing amino acid p-acetyl phenylalanine (pAcPhe) into recombinant proteins expressed in E. coli. Incorporation of pAcPhe is followed by chemoselective reaction of the ketone side chain with a hydroxylamine-functionalized nitroxide to afford the spin-labeled side chain “K1,” and we present two protocols for successful K1 labeling of proteins bearing site-specific pAcPhe. We outline the basic requirements for pAcPhe incorporation and labeling, with an emphasis on practical aspects that must be considered by the researcher if high yields of UAA incorporation and efficient labeling reactions are to be achieved. To this end, we highlight recent advances that have led to increased yields of pAcPhe incorporation, and discuss the use of aniline-based catalysts allowing for facile conjugation of the hydroxylamine spin label under mild reaction conditions. To illustrate the utility of K1 labeling in proteins where traditional cysteine-based SDSL methods are problematic, we site-specifically K1 label the cellular prion protein at two positions in the C-terminal domain and determine the interspin distance using double electron–electron resonance EPR. Recent advances in UAA incorporation and ketone-based bioconjugation, in combination with the commercial availability of all requisite

  10. Genetic Incorporation of the Unnatural Amino Acid p-Acetyl Phenylalanine into Proteins for Site-Directed Spin Labeling.

    PubMed

    Evans, Eric G B; Millhauser, Glenn L

    2015-01-01

    Site-directed spin labeling (SDSL) is a powerful tool for the characterization of protein structure and dynamics; however, its application in many systems is hampered by the reliance on unique and benign cysteine substitutions for the site-specific attachment of the spin label. An elegant solution to this problem involves the use of genetically encoded unnatural amino acids (UAAs) containing reactive functional groups that are chemically orthogonal to those of the 20 amino acids found naturally in proteins. These unique functional groups can then be selectively reacted with an appropriately functionalized spin probe. In this chapter, we detail the genetic incorporation of the ketone-bearing amino acid p-acetyl phenylalanine (pAcPhe) into recombinant proteins expressed in E. coli. Incorporation of pAcPhe is followed by chemoselective reaction of the ketone side chain with a hydroxylamine-functionalized nitroxide to afford the spin-labeled side chain "K1," and we present two protocols for successful K1 labeling of proteins bearing site-specific pAcPhe. We outline the basic requirements for pAcPhe incorporation and labeling, with an emphasis on practical aspects that must be considered by the researcher if high yields of UAA incorporation and efficient labeling reactions are to be achieved. To this end, we highlight recent advances that have led to increased yields of pAcPhe incorporation, and discuss the use of aniline-based catalysts allowing for facile conjugation of the hydroxylamine spin label under mild reaction conditions. To illustrate the utility of K1 labeling in proteins where traditional cysteine-based SDSL methods are problematic, we site-specifically K1 label the cellular prion protein at two positions in the C-terminal domain and determine the interspin distance using double electron-electron resonance EPR. Recent advances in UAA incorporation and ketone-based bioconjugation, in combination with the commercial availability of all requisite reagents

  11. Genetic Incorporation of the Unnatural Amino Acid p-Acetyl Phenylalanine into Proteins for Site-Directed Spin Labeling.

    PubMed

    Evans, Eric G B; Millhauser, Glenn L

    2015-01-01

    Site-directed spin labeling (SDSL) is a powerful tool for the characterization of protein structure and dynamics; however, its application in many systems is hampered by the reliance on unique and benign cysteine substitutions for the site-specific attachment of the spin label. An elegant solution to this problem involves the use of genetically encoded unnatural amino acids (UAAs) containing reactive functional groups that are chemically orthogonal to those of the 20 amino acids found naturally in proteins. These unique functional groups can then be selectively reacted with an appropriately functionalized spin probe. In this chapter, we detail the genetic incorporation of the ketone-bearing amino acid p-acetyl phenylalanine (pAcPhe) into recombinant proteins expressed in E. coli. Incorporation of pAcPhe is followed by chemoselective reaction of the ketone side chain with a hydroxylamine-functionalized nitroxide to afford the spin-labeled side chain "K1," and we present two protocols for successful K1 labeling of proteins bearing site-specific pAcPhe. We outline the basic requirements for pAcPhe incorporation and labeling, with an emphasis on practical aspects that must be considered by the researcher if high yields of UAA incorporation and efficient labeling reactions are to be achieved. To this end, we highlight recent advances that have led to increased yields of pAcPhe incorporation, and discuss the use of aniline-based catalysts allowing for facile conjugation of the hydroxylamine spin label under mild reaction conditions. To illustrate the utility of K1 labeling in proteins where traditional cysteine-based SDSL methods are problematic, we site-specifically K1 label the cellular prion protein at two positions in the C-terminal domain and determine the interspin distance using double electron-electron resonance EPR. Recent advances in UAA incorporation and ketone-based bioconjugation, in combination with the commercial availability of all requisite reagents

  12. Element mobility during pyrite weathering: implications for acid and heavy metal pollution at mining-impacted sites

    NASA Astrophysics Data System (ADS)

    Lu, Long; Wang, Rucheng; Chen, Fanrong; Xue, Jiyue; Zhang, Peihua; Lu, Jianjun

    2005-11-01

    Based on back scattered electron images and electron micro-probe analysis results, four alteration layers, including a transition layer, a reticulated ferric oxide layer, a nubby ferric oxide layer and a cellular ferric oxide layer, were identified in the naturally weathering products of pyrite. These layers represent a progressive alteration sequence of pyrite under weathering conditions. The cellular ferric oxide layer correlates with the strongest weathering phase and results from the dissolution of nubby ferric oxide by acidic porewater. Leaching coefficient was introduced to better express the response of element mobility to the degree of pyrite weathering. Its variation shows that the mobility of S, Co and Bi is stronger than As, Cu and Zn. Sulfur in pyrite is oxidized to sulfuric acid and sulfate that are basically released into to porewater, and heavy metals Co and Bi are evidently released by acid dissolution. As, Cu and Zn are enriched in ferric oxide by adsorption and by co-precipitation, but they would re-release to the environment via desorption or dissolution when porewater pH becomes low enough. Consequently, Co, Bi, As, Cu and Zn may pose a substantial impact on water quality. Considering that metal mobility and its concentration in mine waste are two important factors influencing heavy metal pollution at mining-impacted sites, Bi and Co are more important pollutants in this case.

  13. DNA adducts of aristolochic acid II: total synthesis and site-specific mutagenesis studies in mammalian cells

    PubMed Central

    Attaluri, Sivaprasad; Bonala, Radha R.; Yang, In-Young; Lukin, Mark A.; Wen, Yujing; Grollman, Arthur P.; Moriya, Masaaki; Iden, Charles R.; Johnson, Francis

    2010-01-01

    Aristolochic acids I and II (AA-I, AA-II) are found in all Aristolochia species. Ingestion of these acids either in the form of herbal remedies or as contaminated wheat flour causes a dose-dependent chronic kidney failure characterized by renal tubulointerstitial fibrosis. In ∼50% of these cases, the condition is accompanied by an upper urinary tract malignancy. The disease is now termed aristolochic acid nephropathy (AAN). AA-I is largely responsible for the nephrotoxicity while both AA-I and AA-II are genotoxic. DNA adducts derived from AA-I and AA-II have been isolated from renal tissues of patients suffering from AAN. We describe the total synthesis, de novo, of the dA and dG adducts derived from AA-II, their incorporation site-specifically into DNA oligomers and the splicing of these modified oligomers into a plasmid construct followed by transfection into mouse embryonic fibroblasts. Analysis of the plasmid progeny revealed that both adducts blocked replication but were still partly processed by DNA polymerase(s). Although the majority of coding events involved insertion of correct nucleotides, substantial misincorporation of bases also was noted. The dA adduct is significantly more mutagenic than the dG adduct; both adducts give rise, almost exclusively, to misincorporation of dA, which leads to AL-II-dA→T and AL-II-dG→T transversions. PMID:19854934

  14. Phylogeny of microorganisms populating a thick, subaerial, predominantly lithotrophic biofilm at an extreme acid mine drainage site.

    PubMed

    Bond, P L; Smriga, S P; Banfield, J F

    2000-09-01

    An unusually thick ( approximately 1 cm) slime developed on a slump of finely disseminated pyrite ore within an extreme acid mine drainage site at Iron Mountain, near Redding, Calif. The slime was studied over the period of 1 year. The subaerial form of the slime distinguished it from more typical submerged streamers. Phylogenetic analysis of 16S rRNA genes revealed a diversity of sequences that were mostly novel. Nearest relatives to the majority of sequences came from iron-oxidizing acidophiles, and it appears that iron oxidation is the predominant metabolic characteristic of the organisms in the slime. The most abundant of the 16S rRNA genes detected were from organisms related to Leptospirillum species. The dominant sequence (71% of clones) may represent a new genus. Sequences within the Archaea of the Thermoplasmales lineage were detected. Most of these were only distantly related to known microorganisms. Also, sequences affiliating with Acidimicrobium were detected. Some of these were closely related to "Ferromicrobium acidophilus," and others were affiliated with a lineage only represented by environmental clones. Unexpectedly, sequences that affiliated within the delta subdivision of the Proteobacteria were detected. The predominant metabolic feature of bacteria of this subdivision is anaerobic sulfate or metal reduction. Thus, microenvironments of low redox potential possibly exist in the predominantly oxidizing environments of the slime. These results expand our knowledge of the biodiversity of acid mine drainage environments and extend our understanding of the ecology of extremely acidic systems. PMID:10966399

  15. Modeling solubility and acid-base properties of some amino acids in aqueous NaCl and (CH3)4NCl aqueous solutions at different ionic strengths and temperatures.

    PubMed

    Bretti, Clemente; Giuffrè, Ottavia; Lando, Gabriele; Sammartano, Silvio

    2016-01-01

    New potentiometric experiments have been performed in NaCl and in (CH3)4NCl media, to determine the protonation constants, the protonation enthalpy changes and the solubility of six natural α-amino acids, namely Glycine (Gly), Alanine (Ala), Valine (Val), Leucine (Leu), Serine (Ser) and Phenylalanine (Phe). The aim of the work is the rationalization of the protonation thermodynamics (log [Formula: see text], solubility and [Formula: see text]) in NaCl, determining recommended, tentative or provisional values in selected experimental conditions and to report, for the first time, data in a weak interacting medium, as (CH3)4NCl. Literature data analysis was performed selecting the most reliable values, analyzed together with new data here reported. Significant trends and similarities were observed in the behavior of the six amino acids, and in some cases it was possible to determine common parameters for the ionic strength and temperature dependence. In general, the first protonation step, relative to the amino group, is significantly exothermic (average value is [Formula: see text] = -44.5 ± 0.4 kJ mol(-1) at infinite dilution and T = 298.15 K), and the second, relative to the carboxylate group, is fairly close to zero ([Formula: see text] = -2.5 ± 1.6, same conditions). In both cases, the main contribution to the proton binding reaction is mainly entropic in nature. For phenylalanine and leucine, solubility measurements at different concentrations of supporting electrolyte allowed to determine total and specific solubility values, then used to obtain the Setschenow and the activity coefficients of all the species involved in the protonation equilibria. The values of the first protonation constant in (CH3)4NCl are lower than the corresponding values in NaCl, due to the weak interaction between the deprotonated amino group and (CH3)4N(+). In this light, differences between the protonation functions in NaCl and (CH3)4NCl were used for the quantification

  16. Modeling solubility and acid-base properties of some amino acids in aqueous NaCl and (CH3)4NCl aqueous solutions at different ionic strengths and temperatures.

    PubMed

    Bretti, Clemente; Giuffrè, Ottavia; Lando, Gabriele; Sammartano, Silvio

    2016-01-01

    New potentiometric experiments have been performed in NaCl and in (CH3)4NCl media, to determine the protonation constants, the protonation enthalpy changes and the solubility of six natural α-amino acids, namely Glycine (Gly), Alanine (Ala), Valine (Val), Leucine (Leu), Serine (Ser) and Phenylalanine (Phe). The aim of the work is the rationalization of the protonation thermodynamics (log [Formula: see text], solubility and [Formula: see text]) in NaCl, determining recommended, tentative or provisional values in selected experimental conditions and to report, for the first time, data in a weak interacting medium, as (CH3)4NCl. Literature data analysis was performed selecting the most reliable values, analyzed together with new data here reported. Significant trends and similarities were observed in the behavior of the six amino acids, and in some cases it was possible to determine common parameters for the ionic strength and temperature dependence. In general, the first protonation step, relative to the amino group, is significantly exothermic (average value is [Formula: see text] = -44.5 ± 0.4 kJ mol(-1) at infinite dilution and T = 298.15 K), and the second, relative to the carboxylate group, is fairly close to zero ([Formula: see text] = -2.5 ± 1.6, same conditions). In both cases, the main contribution to the proton binding reaction is mainly entropic in nature. For phenylalanine and leucine, solubility measurements at different concentrations of supporting electrolyte allowed to determine total and specific solubility values, then used to obtain the Setschenow and the activity coefficients of all the species involved in the protonation equilibria. The values of the first protonation constant in (CH3)4NCl are lower than the corresponding values in NaCl, due to the weak interaction between the deprotonated amino group and (CH3)4N(+). In this light, differences between the protonation functions in NaCl and (CH3)4NCl were used for the quantification

  17. Chemical characterization of the surface sites of coal

    SciTech Connect

    Fowkes, F.M.; Lloyd, T.B.; Cole, D.A.

    1990-08-01

    We propose to do experimental studies in four related areas concerning the acid-base properties of coal surfaces; (1) develop high precision flow microcalorimetric methods for determining the concentrations and strengths of the acidic and basic surface sites of coal powders; (2) develop photo-acoustic FTIR and solid-state NMR spectral shift techniques for determination of the concentrations and strengths of acidic and basic surface sites of coal powders; (3) determine the concentrations and strengths of the acidic and basic surface sites of some of the well-characterized coal samples from Argonne National Labs, comparing the coal samples before and after demineralization treatments with HCl and HF; (4) study the effects of surface acidity and basicity on the coal/water interface, with emphasis on the role of interfacial acid-base interactions in the adsorption of ions, surfactants and coal/water slurry stabilizers. A practical application of the new measurements of the acidity and basicity of surface sites will be to exploit acid-base contributions to preferential wettability for optimizing separation of coal pyrites from the organic components. Work this quarter concentrated on flow microcalorimetry for assessing acidity/basicity of coal powders. 8 refs., 9 figs., 1 tab.

  18. Acid potentiation of the capsaicin receptor determined by a key extracellular site.

    PubMed

    Jordt, S E; Tominaga, M; Julius, D

    2000-07-01

    The capsaicin (vanilloid) receptor, VR1, is a sensory neuron-specific ion channel that serves as a polymodal detector of pain-producing chemical and physical stimuli. The response of VR1 to capsaicin or noxious heat is dynamically potentiated by extracellular protons within a pH range encountered during tissue acidosis, such as that associated with arthritis, infarction, tumor growth, and other forms of injury. A molecular determinant for this important physiological activity was localized to an extracellular Glu residue (E600) in the region linking the fifth transmembrane domain with the putative pore-forming region of the channel. We suggest that this residue serves as a key regulatory site of the receptor by setting sensitivity to other noxious stimuli in response to changes in extracellular proton concentration. We also demonstrate that protons, vanilloids, and heat promote channel opening through distinct pathways, because mutations at a second site (E648) selectively abrogate proton-evoked channel activation without diminishing responses to other noxious stimuli. Our findings provide molecular evidence for stimulus-specific steps in VR1 activation and offer strategies for the development of novel analgesic agents. PMID:10859346

  19. Protein engineering of alcohol dehydrogenase--1. Effects of two amino acid changes in the active site of yeast ADH-1.

    PubMed

    Murali, C; Creaser, E H

    1986-01-01

    One of the promises held out by protein engineering is the ability to alter predictably the properties of an enzyme to enable it to find new substrates or catalyse existing substrates more efficiently, such manipulations being of interest both enzymologically and, potentially, industrially. It has been postulated that in yeast alcohol dehydrogenase (YADH-1) certain amino acids such as Trp 93 and Thr 48 constrict the active site due to their bulky side chains and thus impede catalysis of molecules larger than ethanol. To study effects of enlarging the active site we have made two changes into YADH-1, replacing Trp 93 with Phe and Thr 48 with Ser. Kinetic experiments showed that this enzyme had marked increases in reaction velocity for the n-alcohols propanol, butanol, pentanol, hexanol, heptanol, octanol and cinnamyl alcohol compared to the parent, agreeing with the prediction that expanding the active site should facilitate the oxidation of larger alcohols. The substrate affinities were slightly reduced in the altered enzyme, possibly due to its having reduced hydrophobicity at Phe 93.

  20. Isolation and properties of the acid site-specific endonuclease from mature eggs of the sea urchin Strongylocentrotus intermedius

    SciTech Connect

    Sibirtsev, Yu.T.; Konechnyi, A.A.; Rasskazov, V.A.

    1986-01-10

    An acid site-specific endonuclease has been detected in mature sea urchin eggs and cells of embryos at early stages of differentiation. Fractionation with ammonium sulfate, followed by chromatography on columns with DEAE, phosphocellulose, and hydroxyapatite resulted in an 18,000-fold purification. The molecular weight of the enzyme was determined at approx. 29,000, the optimum pH 5.5. The activity of the enzyme does not depend on divalent metal ions, EDTA, ATP, and tRNA, but it is modulated to a substantial degree by NaCl. The maximum rate of cleavage of the DNA supercoil (form I) is observed at 100 mM NaCl. Increasing the NaCl concentration to 350 mM only slightly lowers the rate of cleavage of form I, yielding form II, but entirely suppresses the accumulation of form III. Restriction analysis of the products of enzymatic hydrolysis of Co1E1 and pBR322 DNA showed that at the early stages of hydrolysis the enzyme exhibits pronounced specificity for definite sites, the number of which is 12 for Co1 E1 DNA and 8 sites for pBR322 DNA.

  1. Computational Investigation of Locked Nucleic Acid (LNA) Nucleotides in the Active Sites of DNA Polymerases by Molecular Docking Simulations

    PubMed Central

    Poongavanam, Vasanthanathan; Madala, Praveen K.; Højland, Torben; Veedu, Rakesh N.

    2014-01-01

    Aptamers constitute a potential class of therapeutic molecules typically selected from a large pool of oligonucleotides against a specific target. With a scope of developing unique shorter aptamers with very high biostability and affinity, locked nucleic acid (LNA) nucleotides have been investigated as a substrate for various polymerases. Various reports showed that some thermophilic B-family DNA polymerases, particularly KOD and Phusion DNA polymerases, accepted LNA-nucleoside 5′-triphosphates as substrates. In this study, we investigated the docking of LNA nucleotides in the active sites of RB69 and KOD DNA polymerases by molecular docking simulations. The study revealed that the incoming LNA-TTP is bound in the active site of the RB69 and KOD DNA polymerases in a manner similar to that seen in the case of dTTP, and with LNA structure, there is no other option than the locked C3′-endo conformation which in fact helps better orienting within the active site. PMID:25036012

  2. One-Tube-Only Standardized Site-Directed Mutagenesis: An Alternative Approach to Generate Amino Acid Substitution Collections

    PubMed Central

    Mingo, Janire; Erramuzpe, Asier; Luna, Sandra; Aurtenetxe, Olaia; Amo, Laura; Diez, Ibai; Schepens, Jan T. G.; Hendriks, Wiljan J. A. J.; Cortés, Jesús M.; Pulido, Rafael

    2016-01-01

    Site-directed mutagenesis (SDM) is a powerful tool to create defined collections of protein variants for experimental and clinical purposes, but effectiveness is compromised when a large number of mutations is required. We present here a one-tube-only standardized SDM approach that generates comprehensive collections of amino acid substitution variants, including scanning- and single site-multiple mutations. The approach combines unified mutagenic primer design with the mixing of multiple distinct primer pairs and/or plasmid templates to increase the yield of a single inverse-PCR mutagenesis reaction. Also, a user-friendly program for automatic design of standardized primers for Ala-scanning mutagenesis is made available. Experimental results were compared with a modeling approach together with stochastic simulation data. For single site-multiple mutagenesis purposes and for simultaneous mutagenesis in different plasmid backgrounds, combination of primer sets and/or plasmid templates in a single reaction tube yielded the distinct mutations in a stochastic fashion. For scanning mutagenesis, we found that a combination of overlapping primer sets in a single PCR reaction allowed the yield of different individual mutations, although this yield did not necessarily follow a stochastic trend. Double mutants were generated when the overlap of primer pairs was below 60%. Our results illustrate that one-tube-only SDM effectively reduces the number of reactions required in large-scale mutagenesis strategies, facilitating the generation of comprehensive collections of protein variants suitable for functional analysis. PMID:27548698

  3. The role of amino acid residues in the active site of L-methionine γ-lyase from Pseudomonas putida.

    PubMed

    Fukumoto, Mitsuki; Kudou, Daizou; Murano, Shouko; Shiba, Tomoo; Sato, Dan; Tamura, Takashi; Harada, Shigeharu; Inagaki, Kenji

    2012-01-01

    Cys116, Lys240*, and Asp241* (asterisks indicate residues from the second subunit of the active dimer) at the active site of L-methionine γ-lyase of Pseudomonas putida (MGL_Pp) are highly conserved among heterologous MGLs. In a previous study, we found that substitution of Cys116 for His led to a drastic increase in activity toward L-cysteine and a decrease in that toward L-methionine. In this study, we examined some properties of the C116H mutant by kinetic analysis and 3D structural analysis. We assumed that substitution of Cys116 for His broke the original hydrogen-bond network and that this induced a significant effect of Tyr114 as a general acid catalyst, possibly due to the narrow space in the active site. The C116H mutant acquired a novel β-elimination activity and lead a drastic conformation change in the histidine residue at position 116 by binding the substrate, suggesting that this His residue affects the reaction specificity of C116H. Furthermore, we suggest that Lys240* is important for substrate recognition and structural stability and that Asp241* is also involved in substrate specificity in the elimination reaction. Based on this, we suggest that the hydrogen-bond network among Cys116, Lys240*, and Asp241* contributes to substrate specificity that is, to L-methionine recognition at the active site in MGL_Pp.

  4. One-Tube-Only Standardized Site-Directed Mutagenesis: An Alternative Approach to Generate Amino Acid Substitution Collections.

    PubMed

    Mingo, Janire; Erramuzpe, Asier; Luna, Sandra; Aurtenetxe, Olaia; Amo, Laura; Diez, Ibai; Schepens, Jan T G; Hendriks, Wiljan J A J; Cortés, Jesús M; Pulido, Rafael

    2016-01-01

    Site-directed mutagenesis (SDM) is a powerful tool to create defined collections of protein variants for experimental and clinical purposes, but effectiveness is compromised when a large number of mutations is required. We present here a one-tube-only standardized SDM approach that generates comprehensive collections of amino acid substitution variants, including scanning- and single site-multiple mutations. The approach combines unified mutagenic primer design with the mixing of multiple distinct primer pairs and/or plasmid templates to increase the yield of a single inverse-PCR mutagenesis reaction. Also, a user-friendly program for automatic design of standardized primers for Ala-scanning mutagenesis is made available. Experimental results were compared with a modeling approach together with stochastic simulation data. For single site-multiple mutagenesis purposes and for simultaneous mutagenesis in different plasmid backgrounds, combination of primer sets and/or plasmid templates in a single reaction tube yielded the distinct mutations in a stochastic fashion. For scanning mutagenesis, we found that a combination of overlapping primer sets in a single PCR reaction allowed the yield of different individual mutations, although this yield did not necessarily follow a stochastic trend. Double mutants were generated when the overlap of primer pairs was below 60%. Our results illustrate that one-tube-only SDM effectively reduces the number of reactions required in large-scale mutagenesis strategies, facilitating the generation of comprehensive collections of protein variants suitable for functional analysis. PMID:27548698

  5. Synthesis and Site-Specific Incorporation of Red-Shifted Azobenzene Amino Acids into Proteins.

    PubMed

    John, Alford A; Ramil, Carlo P; Tian, Yulin; Cheng, Gang; Lin, Qing

    2015-12-18

    A series of red-shifted azobenzene amino acids were synthesized in moderate-to-excellent yields via a two-step procedure in which tyrosine derivatives were first oxidized to the corresponding quinonoidal spirolactones followed by ceric ammonium nitrate-catalyzed azo formation with the substituted phenylhydrazines. The resulting azobenzene-alanine derivatives exhibited efficient trans/cis photoswitching upon irradiation with a blue (448 nm) or green (530 nm) LED light. Moreover, nine superfolder green fluorescent protein (sfGFP) mutants carrying the azobenzene-alanine analogues were expressed in E. coli in good yields via amber codon suppression with an orthogonal tRNA/PylRS pair, and one of the mutants showed durable photoswitching with the LED light.

  6. Magic-angle-spinning NMR studies of acid sites in zeolite H-ZSM-5

    SciTech Connect

    Brunner, E.; Ernst, H.; Freude, D.; Froehlich, T.; Hunger, M.; Pfeifer, H. )

    1991-01-01

    {sup 1}H, {sup 13}C, {sup 27}Al, and {sup 29}Si magic-angle-spinning (MAS) NMR was used to elucidate the nature of the catalytic activity of zeolite H-ZSM-5. {sup 1}H MAS NMR of sealed samples after mild hydrothermal dealumination shows that the enhanced activity for n-hexane cracking is not due to an enhanced Bronstead acidity. The concentrations of the various OH groups and aluminous species suggest that the reason for the enhanced catalytic activity is the interaction of the n-hexane molecule with a bridging hydroxyl group and with extra-framework aluminium species, which give rise to the enhanced activity, cannot be easily removed from their positions, and are therefore immobilized by the zeolitic framework.

  7. Site-Selective Binding of Nanoparticles to Double-Stranded DNA via Peptide Nucleic Acid "Invasion"

    SciTech Connect

    Stadler, A.L.; van der Lelie, D.; Sun, D.; Maye, M. M.; Gang, O.

    2011-04-01

    We demonstrate a novel method for by-design placement of nano-objects along double-stranded (ds) DNA. A molecular intercalator, designed as a peptide nucleic acid (PNA)-DNA chimera, is able to invade dsDNA at the PNA-side due to the hybridization specificity between PNA and one of the duplex strands. At the same time, the single-stranded (ss) DNA tail of the chimera, allows for anchoring of nano-objects that have been functionalized with complementary ssDNA. The developed method is applied for interparticle attachment and for the fabrication of particle clusters using a dsDNA template. This method significantly broadens the molecular toolbox for constructing nanoscale systems by including the most conventional not yet utilized DNA motif, double helix DNA.

  8. Neurologic syndrome associated with homozygous mutation at MAG sialic acid binding site.

    PubMed

    Roda, Ricardo H; FitzGibbon, Edmond J; Boucekkine, Houda; Schindler, Alice B; Blackstone, Craig

    2016-08-01

    The MAG gene encodes myelin-associated glycoprotein (MAG), an abundant protein involved in axon-glial interactions and myelination during nerve regeneration. Several members of a consanguineous family with a clinical syndrome reminiscent of Pelizaeus-Merzbacher disease and demyelinating leukodystrophy on brain MRI were recently found to harbor a homozygous missense p.Ser133Arg MAG mutation. Here, we report two brothers from a nonconsanguineous family afflicted with progressive cognitive impairment, neuropathy, ataxia, nystagmus, and gait disorder. Exome sequencing revealed the homozygous missense mutation p.Arg118His in MAG. This Arg118 residue in immunoglobulin domain 1 is critical for sialic acid binding, providing a compelling mechanistic basis for disease pathogenesis. PMID:27606346

  9. A Wood-Waste Cover Prevents Sulphide Oxidation and Treats Acid Effluents at the East-Sullivan Mine Site

    NASA Astrophysics Data System (ADS)

    Germain, D.; Tassé, N.; Cyr, J.

    2004-05-01

    At the East Sullivan site, wood wastes covering the abandoned mine tailings impoundment prevent sulphide oxidation by creating an anoxic environment. The addition of coarse ligneous wastes favours infiltration, resulting in a water table rise. This maintains most tailings saturated and thus provides an additional protection against sulphide oxidation. Moreover, high infiltration allows a more rapid flushing of acid prone groundwater generated prior to the cover placement. Finally, the pore-waters under the cover are characterized by a strong reducing potential and high alkalinity. These conditions favour sulphate reduction and base metal precipitation as sulphides and carbonates. The restoration strategy capitalized on the alkaline and reductive properties of the waters underlying the wood-waste cover. An original treatment of acid effluents, based on the recirculation of water discharging around the impoundment through the organic cover, was implemented in 1998. In 2003, the total volume of water treated was 725 000 m3. Data gathered near the dispersal zone show that despite dispersing acid water, the groundwater pH decreases by only one unit from 7 to 6, during the recirculation period: May to October. However, alkalinity decreases from 800 to 100 mg/L-CaCO3. But it is back up to 800 mg/L the following spring, thanks to sulphate reduction. Fe2+ concentrations near the dispersal zone are maintained below 2 mg/L. Evolution of the iron mass in the surface waters suggests that the contaminated groundwater flush is completed in the north and west sectors of the impoundment; the east and south ones are expected to be recovered within 3 to 4 years. A wood-waste cover, besides limiting sulphide oxidation, can fill the role of alkaline reducing barrier for the treatment of these acidogenic waters, until a balance between acidity and alkalinity in the effluent is reached.

  10. Schiff base structured acid-base cooperative dual sites in an ionic solid catalyst lead to efficient heterogeneous knoevenagel condensations.

    PubMed

    Zhang, Mingjue; Zhao, Pingping; Leng, Yan; Chen, Guojian; Wang, Jun; Huang, Jun

    2012-10-01

    An acid-base bifunctional ionic solid catalyst [PySaIm](3)PW was synthesized by the anion exchange of the ionic-liquid (IL) precursor 1-(2-salicylaldimine)pyridinium bromide ([PySaIm]Br) with the Keggin-structured sodium phosphotungstate (Na(3) PW). The catalyst was characterized by FTIR, UV/Vis, XRD, SEM, Brunauer-Emmett-Teller (BET) theory, thermogravimetric analysis, (1)H NMR spectroscopy, ESI-MS, elemental analysis, and melting points. Together with various counterparts, [PySaIm](3)PW was evaluated in Knoevenagel condensation under solvent and solvent-free conditions. The Schiff base structure attached to the IL cation of [PySaIm](3)PW involves acidic salicyl hydroxyl and basic imine, and provides a controlled nearby position for the acid-base dual sites. The high melting and insoluble properties of [PySaIm](3)PW are relative to the large volume and high valence of PW anions, as well as the intermolecular hydrogen-bonding networks among inorganic anions and IL cations. The ionic solid catalyst [PySaIm](3)PW leads to heterogeneous Knoevenagel condensations. In solvent-free condensation of benzaldehyde with ethyl cyanoacetate, it exhibits a conversion of 95.8 % and a selectivity of 100 %; the conversion is even much higher than that (78.2 %) with ethanol as a solvent. The solid catalyst has a convenient recoverability with only a slight decrease in conversion following subsequent recyclings. Furthermore, the new catalyst is highly applicable to many substrates of aromatic aldehydes with activated methylene compounds. On the basis of the characterization and reaction results, a unique acid-base cooperative mechanism within a Schiff base structure is proposed and discussed, which thoroughly explains not only the highly efficient catalytic performance of [PySaIm](3)PW, but also the lower activities of various control catalysts.

  11. Characterization of cyclo-Acetoacetyl-L-Tryptophan Dimethylallyltransferase in Cyclopiazonic Acid Biosynthesis: Substrate Promiscuity and Site Directed Mutagenesis Studies

    PubMed Central

    Liu, Xinyu; Walsh, Christopher T.

    2009-01-01

    The fungal neurotoxin α-cyclopiazonic acid (CPA), a nanomolar inhibitor of Ca2+-ATPase with a unique pentacyclic indole tetramic acid scaffold is assembled by a three enzyme pathway CpaS, CpaD and CpaO in Aspergillus sp. We recently characterized the first pathway-specific enzyme CpaS, a hybrid two module polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS) that generates cyclo-acetoacetyl-L-tryptophan (cAATrp). Here we report the characterization of the second pathway-specific enzyme CpaD that regiospecifically dimethylallylates cAATrp to form β-cyclopiazonic acid. By exploring the tryptophan and tetramate moieties of cAATrp, we demonstrate that CpaD discriminates against free Trp but accepts tryptophan-containing thiohydantoins, diketopiperazines and linear peptides as substrates for C4-prenylation and also acts as regiospecific O-dimethylallyltransferase (DMAT) on a tyrosine-derived tetramic acid. Comparative evaluation of CpaDs from A. oryzae RIB40 and A. flavus NRRL3357 indicated the importance of the N-terminal region for its activity. Sequence alignment of CpaD with eleven homologous fungal Trp-DMATs revealed five regions of conservation suggesting the presense of critical motifs that could be diagonostic for discovering additional Trp-DMATs. Subsequent site-directed mutagenesis studies identified five polar/charged residues and five tyrosine residues within these motifs that are critical for CpaD activity. This motif characerization will enable a gene probe-based approach to discover additional biosynthetic Trp-DMATs. PMID:19877600

  12. Direct conversion of bio-ethanol to isobutene on nanosized Zn(x)Zr(y)O(z) mixed oxides with balanced acid-base sites.

    PubMed

    Sun, Junming; Zhu, Kake; Gao, Feng; Wang, Chongmin; Liu, Jun; Peden, Charles H F; Wang, Yong

    2011-07-27

    We report the design and synthesis of nanosized Zn(x)Zr(y)O(z) mixed oxides for direct and high-yield conversion of bio-ethanol to isobutene (~83%). ZnO is addded to ZrO(2) to selectively passivate zirconia's strong Lewis acidic sites and weaken Brönsted acidic sites, while simultaneously introducing basicity. As a result, the undesired reactions of bio-ethanol dehydration and acetone polymerization/coking are suppressed. Instead, a surface basic site-catalyzed ethanol dehydrogenation to acetaldehyde, acetaldehyde to acetone conversion via a complex pathway including aldol-condensation/dehydrogenation, and a Brönsted acidic site-catalyzed acetone-to-isobutene reaction pathway dominates on the nanosized Zn(x)Zr(y)O(z) mixed oxide catalyst, leading to a highly selective process for direct conversion of bio-ethanol to isobutene. PMID:21682296

  13. A Mutational Analysis of Active Site Residues in trans-3-Chloroacrylic Acid Dehalogenase

    PubMed Central

    Poelarends, Gerrit J.; Serrano, Hector; Huddleston, Jamison P.; Johnson, William H.; Whitman, Christian P.

    2013-01-01

    trans -3-Chloroacrylic acid dehalogenase (CaaD) catalyzes the hydrolytic dehalogenation of trans-3-haloacrylates to yield malonate semialdehyde by a mechanism utilizing βPro-1, αArg-8, αArg-11, and αGlu-52. These residues are implicated in a promiscuous hydratase activity where 2-oxo-3-pentynoate is processed to acetopyruvate. The roles of three nearby residues (βAsn-39, αPhe-39, and αPhe-50) are unexplored. Mutants were constructed at these positions (βN39A, αF39A, αF39T, αF50A and αF50Y) and kinetic parameters determined along with those of the αR8K and αR11K mutants. Analysis indicates that αArg-8, αArg-11, and βAsn-39 are critical for dehalogenase activity whereas αArg-11 and αPhe-50 are critical for hydratase activity. Docking studies suggest structural bases for these observations. PMID:23851010

  14. Acid mine drainage risks - A modeling approach to siting mine facilities in Northern Minnesota USA

    NASA Astrophysics Data System (ADS)

    Myers, Tom

    2016-02-01

    Most watershed-scale planning for mine-caused contamination concerns remediation of past problems while future planning relies heavily on engineering controls. As an alternative, a watershed scale groundwater fate and transport model for the Rainy Headwaters, a northeastern Minnesota watershed, has been developed to examine the risks of leaks or spills to a pristine downstream watershed. The model shows that the risk depends on the location and whether the source of the leak is on the surface or from deeper underground facilities. Underground sources cause loads that last longer but arrive at rivers after a longer travel time and have lower concentrations due to dilution and attenuation. Surface contaminant sources could cause much more short-term damage to the resource. Because groundwater dominates baseflow, mine contaminant seepage would cause the most damage during low flow periods. Groundwater flow and transport modeling is a useful tool for decreasing the risk to downgradient sources by aiding in the placement of mine facilities. Although mines are located based on the minerals, advance planning and analysis could avoid siting mine facilities where failure or leaks would cause too much natural resource damage. Watershed scale transport modeling could help locate the facilities or decide in advance that the mine should not be constructed due to the risk to downstream resources.

  15. Site-directed mutagenesis of tobacco anionic peroxidase: Effect of additional aromatic amino acids on stability and activity.

    PubMed

    Poloznikov, A A; Zakharova, G S; Chubar, T A; Hushpulian, D M; Tishkov, V I; Gazaryan, I G

    2015-08-01

    Tobacco anionic peroxidase (TOP) is known to effectively catalyze luminol oxidation without enhancers, in contrast to horseradish peroxidase (HRP). To pursue structure-activity relationship studies for TOP, two amino acids have been chosen for mutation, namely Thr151, close to the heme plane, and Phe140 at the entrance to the active site pocket. Three mutant forms TOP F140Y, T151W and F140Y/T151W have been expressed in Escherichia coli, and reactivated to yield active enzymes. Single-point mutations introducing additional aromatic amino acid residues at the surface of TOP exhibit a significant effect on the enzyme catalytic activity and stability as judged by the results of steady-state and transient kinetics studies. TOP T151W is up to 4-fold more active towards a number of aromatic substrates including luminol, whereas TOP F140Y is 2-fold more stable against thermal inactivation and 8-fold more stable in the reaction course. These steady-state observations have been rationalized with the help of transient kinetic studies on the enzyme reaction with hydrogen peroxide in a single turnover regime. The stopped-flow data reveal (a) an increased stability of F140Y Compound I towards hydrogen peroxide, and thus, a higher operational stability as compared to the wild-type enzyme, and (b) a lesser leakage of oxidative equivalents from TOP T151W Compound I resulting in the increased catalytic activity. The results obtained show that TOP unique properties can be further improved for practical applications by site-directed mutagenesis.

  16. Changes in polysialic acid expression on myeloid cells during differentiation and recruitment to sites of inflammation: Role in phagocytosis

    PubMed Central

    Stamatos, Nicholas M; Zhang, Lei; Jokilammi, Anne; Finne, Jukka; Chen, Wilbur H; El-Maarouf, Abderrahman; Cross, Alan S; Hankey, Kim G

    2014-01-01

    Polysialic acid (polySia) is a unique linear homopolymer of α2,8-linked sialic acid that has been studied extensively as a posttranslational modification of neural cell adhesion molecule in the central nervous system. Only two proteins are known to be polysialylated in cells of the immune system: CD56 on human natural killer cells and murine bone marrow (BM) leukocytes, and neuropilin-2 (NRP-2) on dendritic cells (DCs). We tested the hypothesis that polySia expression is regulated during maturation and migration of leukocytes and plays a role in functional activity. Using wild-type and NCAM−/− mice, we show that BM neutrophils express only polysialylated CD56, whereas a subset of BM monocytes expresses polysialylated CD56 and/or another polysialylated protein(s). We demonstrate that polysialylated CD56 expression is progressively down-regulated in wild-type monocytes and monocyte-derived cells during migration from BM through peripheral blood to pulmonary and peritoneal sites of inflammation. Freshly isolated monocyte-derived peritoneal macrophages are devoid of polySia yet re-express polySia on NRP-2 and an additional protein(s) after maintenance in culture. Removal of polySia from these cells enhances phagocytosis of Klebsiella pneumoniae, suggesting that down-regulation of polySia on macrophages facilitates bacterial clearance. Using wild-type and NRP-2−/− mice, we demonstrate that NRP-2 and an additional protein(s) are polysialylated by ST8 SiaIV in BM-derived DCs. We conclude that polySia expression in monocyte-derived cells is dynamically regulated by ST8 SiaIV activity and by expression of carrier proteins during recruitment to sites of inflammation and influences cellular interactions with microbes, contributing to innate and adaptive immune responses. PMID:24865221

  17. A mass-spectrometric method for the estimation of the ratio of gamma-carboxyglutamic acid to glutamic acid at specific sites in proteins. Application to the N-terminal region of bovine prothrombin.

    PubMed

    Rose, K; Priddle, J D; Offord, R E; Esnouf, M P

    1980-04-01

    When a polypeptide containing gamma-carboxyglutamic acid is decarboxylated in 2H2O, residue of (gamma gamma-2H2)glutamic acid are formed. Subsequent proteolytic digestion produces peptides which contain at each site 2H2-substituted and unsubstituted glutamic acid in the same ratio as existed for gramma-carboxy-substitution. The peptides may be identified and this ratio determined by combined gas chromatography--mass spectrometry. We also discuss decarboxylation in 3H2O followed by amino-acid analysis and Edman degradation.

  18. Newly identified essential amino acid residues affecting Δ8-sphingolipid desaturase activity revealed by site-directed mutagenesis.

    PubMed

    Li, Shu-Fen; Song, Li-Ying; Zhang, Guo-Jun; Yin, Wei-Bo; Chen, Yu-Hong; Wang, Richard R-C; Hu, Zan-Min

    2011-12-01

    In order to identify amino acid residues crucial for the enzymatic activity of Δ(8)-sphingolipid desaturases, a sequence comparison was performed among Δ(8)-sphingolipid desaturases and Δ(6)-fatty acid desaturases from various plants. In addition to the known conserved cytb(5) (cytochrome b(5)) HPGG motif and three conserved histidine boxes, they share additional 15 completely conserved residues. A series of site-directed mutants were generated using our previously isolated Δ(8)-sphingolipid desaturase gene from Brassica rapa to evaluate the importance of these residues to the enzyme function. The mutants were functionally characterized by heterologous expression in yeast, allowing the identification of the products of the enzymes. The results revealed that residues H63, N203, D208, D210, and G368 were obligatorily required for the enzymatic activity, and substitution of the residues F59, W190, W345, L369 and Q372 markedly decreased the enzyme activity. Among them, replacement of the residues W190, L369 and Q372 also has significant influence on the ratio of the two enzyme products. Information obtained in this work provides the molecular basis for the Δ(8)-sphingolipid desaturase activity and aids in our understanding of the structure-function relationships of the membrane-bound desaturases.

  19. The active site of purple acid phosphatase from sweet potatoes (Ipomoea batatas) metal content and spectroscopic characterization.

    PubMed

    Durmus, A; Eicken, C; Sift, B H; Kratel, A; Kappl, R; Hüttermann, J; Krebs, B

    1999-03-01

    Purple acid phosphatase from sweet potatoes Ipomoea batatas (spPAP) has been purified to homogeneity and characterized using spectroscopic investigations. Matrix-assisted laser desorption/ionization mass spectrometry analysis revealed a molecular mass of approximately 112 kDa. The metal content was determined by X-ray fluorescence using synchrotron radiation. In contrast to previous studies it is shown that spPAP contains a Fe(III)-Zn(II) center in the active site as previously determined for the purple acid phosphatase from red kidney bean (kbPAP). Moreover, an alignment of the amino acid sequences suggests that the residues involved in metal-binding are identical in both plant PAPs. Tyrosine functions as one of the ligands for the chromophoric Fe(III). Low temperature EPR spectra of spPAP show a signal near g = 4.3, characteristic for high-spin Fe(III) in a rhombic environment. The Tyr-Fe(III) charge transfer transition and the EPR signal are both very sensitive to changes in pH. The pH dependency strongly suggests the presence of an ionizable group with a pKa of 4.7, arising from an aquo ligand coordinated to Fe(III). EPR and UV/visible studies of spPAP in the presence of the inhibitors phosphate or arsenate suggest that both anions bind to Fe(III) in the binuclear center replacing the coordinated water or hydroxide ligand necessary for hydrolysis. The conserved histidine residues of spPAP corresponding to His202 and His296 in kbPAP probably interact in catalysis. PMID:10102999

  20. Vesiculoviral matrix (M) protein occupies nucleic acid binding site at nucleoporin pair (Rae1 • Nup98).

    PubMed

    Quan, Beili; Seo, Hyuk-Soo; Blobel, Günter; Ren, Yi

    2014-06-24

    mRNA export factor 1 (Rae1) and nucleoporin 98 (Nup98) are host cell targets for the matrix (M) protein of vesicular stomatitis virus (VSV). How Rae1 functions in mRNA export and how M protein targets both Rae1 and Nup98 are not understood at the molecular level. To obtain structural insights, we assembled a 1:1:1 complex of M•Rae1•Nup98 and established a crystal structure at 3.15-Å resolution. We found that the M protein contacts the Rae1•Nup98 heterodimer principally by two protrusions projecting from the globular domain of M like a finger and thumb. Both projections clamp to the side of the β-propeller of Rae1, with the finger also contacting Nup98. The most prominent feature of the finger is highly conserved Methionine 51 (Met51) with upstream and downstream acidic residues. The complementary surface on Rae1 displays a deep hydrophobic pocket, into which Met51 fastens like a bolt, and a groove of basic residues on either side, which bond to the acidic residues of the finger. Notably, the M protein competed for in vitro binding of various oligonucleotides to Rae1•Nup98. We localized this competing activity of M to its finger using a synthetic peptide. Collectively, our data suggest that Rae1 serves as a binding protein for the phosphate backbone of any nucleic acid and that the finger of M mimics this ligand. In the context of mRNA export, we propose that a given mRNA segment, after having been deproteinated by helicase, is transiently reproteinated by Nup98-tethered Rae1. We suggest that such repetitive cycles provide cytoplasmic stopover sites required for ratcheting mRNA across the nuclear pore.

  1. The active site of purple acid phosphatase from sweet potatoes (Ipomoea batatas) metal content and spectroscopic characterization.

    PubMed

    Durmus, A; Eicken, C; Sift, B H; Kratel, A; Kappl, R; Hüttermann, J; Krebs, B

    1999-03-01

    Purple acid phosphatase from sweet potatoes Ipomoea batatas (spPAP) has been purified to homogeneity and characterized using spectroscopic investigations. Matrix-assisted laser desorption/ionization mass spectrometry analysis revealed a molecular mass of approximately 112 kDa. The metal content was determined by X-ray fluorescence using synchrotron radiation. In contrast to previous studies it is shown that spPAP contains a Fe(III)-Zn(II) center in the active site as previously determined for the purple acid phosphatase from red kidney bean (kbPAP). Moreover, an alignment of the amino acid sequences suggests that the residues involved in metal-binding are identical in both plant PAPs. Tyrosine functions as one of the ligands for the chromophoric Fe(III). Low temperature EPR spectra of spPAP show a signal near g = 4.3, characteristic for high-spin Fe(III) in a rhombic environment. The Tyr-Fe(III) charge transfer transition and the EPR signal are both very sensitive to changes in pH. The pH dependency strongly suggests the presence of an ionizable group with a pKa of 4.7, arising from an aquo ligand coordinated to Fe(III). EPR and UV/visible studies of spPAP in the presence of the inhibitors phosphate or arsenate suggest that both anions bind to Fe(III) in the binuclear center replacing the coordinated water or hydroxide ligand necessary for hydrolysis. The conserved histidine residues of spPAP corresponding to His202 and His296 in kbPAP probably interact in catalysis.

  2. Vesiculoviral matrix (M) protein occupies nucleic acid binding site at nucleoporin pair (Rae1∙Nup98)

    SciTech Connect

    Quan, Beili; Seo, Hyuk-Soo; Blobel, Günter; Ren, Yi

    2014-07-01

    mRNA export factor 1 (Rae1) and nucleoporin 98 (Nup98) are host cell targets for the matrix (M) protein of vesicular stomatitis virus (VSV). How Rae1 functions in mRNA export and how M protein targets both Rae1 and Nup98 are not understood at the molecular level. To obtain structural insights, we assembled a 1:1:1 complex of M•Rae1•Nup98 and established a crystal structure at 3.15-Å resolution. We found that the M protein contacts the Rae1•Nup98 heterodimer principally by two protrusions projecting from the globular domain of M like a finger and thumb. Both projections clamp to the side of the β-propeller of Rae1, with the finger also contacting Nup98. The most prominent feature of the finger is highly conserved Methionine 51 (Met51) with upstream and downstream acidic residues. The complementary surface on Rae1 displays a deep hydrophobic pocket, into which Met51 fastens like a bolt, and a groove of basic residues on either side, which bond to the acidic residues of the finger. Notably, the M protein competed for in vitro binding of various oligonucleotides to Rae1•Nup98. We localized this competing activity of M to its finger using a synthetic peptide. Collectively, our data suggest that Rae1 serves as a binding protein for the phosphate backbone of any nucleic acid and that the finger of M mimics this ligand. In the context of mRNA export, we propose that a given mRNA segment, after having been deproteinated by helicase, is transiently reproteinated by Nup98-tethered Rae1. We suggest that such repetitive cycles provide cytoplasmic stopover sites required for ratcheting mRNA across the nuclear pore.

  3. Vesiculoviral matrix (M) protein occupies nucleic acid binding site at nucleoporin pair (Rae1•Nup98)

    PubMed Central

    Quan, Beili; Seo, Hyuk-Soo; Blobel, Günter; Ren, Yi

    2014-01-01

    mRNA export factor 1 (Rae1) and nucleoporin 98 (Nup98) are host cell targets for the matrix (M) protein of vesicular stomatitis virus (VSV). How Rae1 functions in mRNA export and how M protein targets both Rae1 and Nup98 are not understood at the molecular level. To obtain structural insights, we assembled a 1:1:1 complex of M•Rae1•Nup98 and established a crystal structure at 3.15-Å resolution. We found that the M protein contacts the Rae1•Nup98 heterodimer principally by two protrusions projecting from the globular domain of M like a finger and thumb. Both projections clamp to the side of the β-propeller of Rae1, with the finger also contacting Nup98. The most prominent feature of the finger is highly conserved Methionine 51 (Met51) with upstream and downstream acidic residues. The complementary surface on Rae1 displays a deep hydrophobic pocket, into which Met51 fastens like a bolt, and a groove of basic residues on either side, which bond to the acidic residues of the finger. Notably, the M protein competed for in vitro binding of various oligonucleotides to Rae1•Nup98. We localized this competing activity of M to its finger using a synthetic peptide. Collectively, our data suggest that Rae1 serves as a binding protein for the phosphate backbone of any nucleic acid and that the finger of M mimics this ligand. In the context of mRNA export, we propose that a given mRNA segment, after having been deproteinated by helicase, is transiently reproteinated by Nup98-tethered Rae1. We suggest that such repetitive cycles provide cytoplasmic stopover sites required for ratcheting mRNA across the nuclear pore. PMID:24927547

  4. Binding site multiplicity with fatty acid ligands: implications for the regulation of PKR kinase autophosphorylation with palmitate.

    PubMed

    Fang, Liang; Cho, Hyun Ju; Chan, Christina; Feig, Michael

    2014-10-01

    Saturated long chain-free fatty acids (FFAs), especially palmitate, have been implicated in apoptosis by inhibiting the activity of PKR (double-stranded RNA-dependent protein kinase). We recently found evidence that palmitate interacts directly with the kinase domain of PKR, subsequently inhibiting the autophosphorylation of PKR. To investigate the interactions of palmitate with PKR and its effects on PKR autophosphorylation, we performed extensive unbiased MD simulations combined with biochemical and biophysical experiments. The simulations predict multiple putative binding sites of palmitate on both the phosphorylated and unphosphorylated PKR with similar binding affinities. Ligand-protein interactions involving a large variety of different binding modes challenge the conventional view of highly specific, single binding sites. Key interactions of palmitate involve the αC-helix of PKR, especially near residue R307. Experimental mutation of R307 was found to affect palmitate binding and reduce its inhibitory effect. Based on this study a new allosteric mechanism is proposed where palmitate binding to the αC-helix prevents the inactive-to-active transition of PKR and subsequently reduces its ability to autophosphorylate.

  5. Experimental and computational investigation of acetic acid deoxygenation over oxophilic molybdenum carbide: Surface chemistry and active site identity

    DOE PAGES

    Schaidle, Joshua A.; Blackburn, Jeffrey; Farberow, Carrie A.; Nash, Connor; Steirer, K. Xerxes; Clark, Jared; Robichaud, David J.; Ruddy, Daniel A.

    2016-01-21

    Ex situ catalytic fast pyrolysis (CFP) is a promising route for producing fungible biofuels; however, this process requires bifunctional catalysts that favor C–O bond cleavage, activate hydrogen at near atmospheric pressure and high temperature (350–500 °C), and are stable under high-steam, low hydrogen-to-carbon environments. Recently, early transition-metal carbides have been reported to selectively cleave C–O bonds of alcohols, aldehydes, and oxygenated aromatics, yet there is limited understanding of the metal carbide surface chemistry under reaction conditions and the identity of the active sites for deoxygenation. In this study, we evaluated molybdenum carbide (Mo2C) for the deoxygenation of acetic acid, anmore » abundant component of biomass pyrolysis vapors, under ex situ CFP conditions, and we probed the Mo2C surface chemistry, identity of the active sites, and deoxygenation pathways using in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), X-ray photoelectron spectroscopy (XPS), and density functional theory (DFT) calculations.« less

  6. Multiplex Detection of Functional G Protein-Coupled Receptors Harboring Site-Specifically Modified Unnatural Amino Acids

    PubMed Central

    2015-01-01

    We developed a strategy for identifying positions in G protein-coupled receptors that are amenable to bioorthogonal modification with a peptide epitope tag under cell culturing conditions. We introduced the unnatural amino acid p-azido-l-phenylalanine (azF) into human CC chemokine receptor 5 (CCR5) at site-specific amber codon mutations. We then used strain-promoted azide–alkyne [3+2] cycloaddition to label the azF-CCR5 variants with a FLAG peptide epitope-conjugated aza-dibenzocyclooctyne (DBCO) reagent. A microtiter plate-based sandwich fluorophore-linked immunosorbent assay was used to probe simultaneously the FLAG epitope and the receptor using infrared dye-conjugated antibodies so that the extent of DBCO incorporation, corresponding nominally to labeling efficiency, could be quantified ratiometrically. The extent of incorporation of DBCO at the various sites was evaluated in the context of a recent crystal structure of maraviroc-bound CCR5. We observed that labeling efficiency varied dramatically depending on the topological location of the azF in CCR5. Interestingly, position 109 in transmembrane helix 3, located in a hydrophobic cavity on the extracellular side of the receptor, was labeled most efficiently. Because the bioorthogonal labeling and detection strategy described might be used to introduce a variety of different peptide epitopes or fluorophores into engineered expressed receptors, it might prove to be useful for a wide range of applications, including single-molecule detection studies of receptor trafficking and signaling mechanism. PMID:25524496

  7. Structural model for gamma-aminobutyric acid receptor noncompetitive antagonist binding: widely diverse structures fit the same site.

    PubMed

    Chen, Ligong; Durkin, Kathleen A; Casida, John E

    2006-03-28

    Several major insecticides, including alpha-endosulfan, lindane, and fipronil, and the botanical picrotoxinin are noncompetitive antagonists (NCAs) for the GABA receptor. We showed earlier that human beta(3) homopentameric GABA(A) receptor recognizes all of the important GABAergic insecticides and reproduces the high insecticide sensitivity and structure-activity relationships of the native insect receptor. Despite large structural diversity, the NCAs are proposed to fit a single binding site in the chloride channel lumen lined by five transmembrane 2 segments. This hypothesis is examined with the beta(3) homopentamer by mutagenesis, pore structure studies, NCA binding, and molecular modeling. The 15 amino acids in the cytoplasmic half of the pore were mutated to cysteine, serine, or other residue for 22 mutants overall. Localization of A-1'C, A2'C, T6'C, and L9'C (index numbers for the transmembrane 2 region) in the channel lumen was established by disulfide cross-linking. Binding of two NCA radioligands [(3)H]1-(4-ethynylphenyl)-4-n-propyl-2,6,7-trioxabicyclo[2.2.2]octane and [(3)H] 3,3-bis-trifluoromethyl-bicyclo[2,2,1]heptane-2,2-dicarbonitrile was dramatically reduced with 8 of the 15 mutated positions, focusing attention on A2', T6', and L9' as proposed binding sites, consistent with earlier mutagenesis studies. The cytoplasmic half of the beta3 homopentamer pore was modeled as an alpha-helix. The six NCAs listed above plus t-butylbicyclophosphorothionate fit the 2' to 9' pore region forming hydrogen bonds with the T6' hydroxyl and hydrophobic interactions with A2', T6', and L9' alkyl substituents, thereby blocking the channel. Thus, widely diverse NCA structures fit the same GABA receptor beta subunit site with important implications for insecticide cross-resistance and selective toxicity between insects and mammals.

  8. Encapsulating Metal Clusters and Acid Sites within Small Voids: Synthetic Strategies and Catalytic Consequences

    NASA Astrophysics Data System (ADS)

    Goel, Sarika

    The selective encapsulation of metal clusters within zeolites can be used to prepare clusters that are uniform in diameter and to protect them against sintering and contact with feed impurities, while concurrently allowing active sites to select reactants based on their molecular size, thus conferring enzyme-like specificity to chemical catalysis. The apertures in small and medium-pore zeolites preclude the use of post-synthetic protocols to encapsulate the relevant metal precursors because cationic or anionic precursors with their charge-balancing double layer and gaseous complexes cannot diffuse through their windows or channels. We have developed general strategies to encapsulate metal clusters within small-pore zeolites by using metal precursors stabilized by ammonia or organic amine ligands, which stabilize metal precursors against their premature precipitation at the high temperature and pH conditions required for the hydrothermal synthesis of the target zeolite structures and favor interactions between metal precursors and incipient aluminosilicate nuclei during the self-assembly of microporous frameworks. When synthesis temperatures were higher than 400 K, available ligands were unable to prevent the premature precipitation of the metal precursors. In such cases, encapsulation was achieved instead via interzeolite transformations after successfully encapsulating metal precursors or clusters via post-synthesis exchange or ligand protection into parent zeolites and subsequently converting them into the target structures while retaining the encapsulated clusters or precursors. Such strategies led to the successful selective encapsulation of a wide range of metal clusters (Pt, Pd, Ru, Rh, Ir, Re, and Ag) within small-pore (SOD (sodalite), LTA (Linde type A (zeolite A)), GIS (gismondine), and ANA (analcime)) and medium-pore (MFI (ZSM-5)) zeolites. These protocols provide novel and diverse mechanism-based strategies for the design of catalysts with protected

  9. SORPTION OF CU AND ZN TO KAOLINITE AND IRON OXIDE: EFFECTS OF HUMIC ACID AND IONIC STRENGTH AND IMPLICATIONS FOR STORMWATER RUNOFF

    EPA Science Inventory

    Heavy metals are common pollutants in wet weather flows and urban waterways. Changes in ionic strength, whether from mixing with saline waters, road salt, or from the large osmotic adjustment needed for the Microtox toxicity assay, affect the aqueous chemistry of stormwater runof...

  10. Hydrogeology and ground-water quality of the Chromic Acid Pit site, US Army Air Defense Artillery Center and Fort Bliss, El Paso, Texas

    USGS Publications Warehouse

    Abeyta, Cynthia G.; Thomas, C.L.

    1996-01-01

    The Chromic Acid Pit site is an inactive waste disposal site that is regulated by the Resource Conservation and Recovery Act of 1976. The 2.2-cubic-yard cement-lined pit was operated from 1980 to 1983 by a contractor to the U.S. Army Air Defense Artillery Center and Fort Bliss. The pit, located on the Fort Bliss military reservation, in El Paso, Texas, was used for disposal and evaporation of chromic acid waste generated from chrome plating operations. The site was certified closed in 1989 and the Texas Natural Resources Conservation Commission issued Permit Number HW-50296 (U.S. Environmental Protection Agency Permit Number TX4213720101), which approved and implemented post-closure care for the Chromic Acid Pit site. In accordance with an approved post-closure plan, the U.S. Geological Survey is cooperating with the U.S. Army in evaluating hydrogeologic conditions and ground- water quality at the site. One upgradient and two downgradient ground-water monitoring wells were installed adjacent to the chromic acid pit by a private contractor. Quarterly ground-water sampling of these wells by the U.S. Geological Survey began in December 1993. The Chromic Acid Pit site is situated in the Hueco Bolson intermontane valley. The Hueco Bolson is a primary source of ground water in the El Paso area. City of El Paso and U.S. Army water-supply wells are located on all sides of the study area and are completed 600 to more than 1,200 feet below land surface. The ground-water level in the area of the Chromic Acid Pit site has declined about 25 feet from 1982 to 1993. Depth to water at the Chromic Acid Pit site in September 1994 was about 284 feet below land surface; ground-water flow is to the southeast. Ground-water samples collected from monitoring wells at the Chromic Acid Pit site contained dissolved-solids concentrations of 442 to 564 milligrams per liter. Nitrate as nitrogen concentrations ranged from 2.1 to 2.7 milligrams per liter; nitrite plus nitrate as nitrogen

  11. Molecular simulations of β-lactoglobulin complexed with fatty acids reveal the structural basis of ligand affinity to internal and possible external binding sites.

    PubMed

    Evoli, Stefania; Guzzi, Rita; Rizzuti, Bruno

    2014-10-01

    The interaction of saturated fatty acids of different length (C8:0 to C18:0) with β-lactoglobulin (βLG) was investigated by molecular dynamics simulation and docking approaches. The results show that the presence of such ligands in the hydrophobic central cavity of βLG, known as the protein calyx, determines an enhancement of atomic fluctuations compared with the unliganded form, especially for loops at the entrance of the binding site. Concerted motions are evidenced for protein regions that could favor the binding of ligands. The mechanism of anchoring of fatty acids of different length is similar for the carboxylate head-group, through electrostatic interactions with the side chains of Lys60/Lys69. The key protein residues to secure the hydrocarbon chain are Phe105/Met107, which adapt their conformation upon ligand binding. In particular, Phe105 provides an additional hydrophobic clamp only for the tail of the two fatty acids with the longest chains, palmitic, and stearic acid, which are known to bind βLG with a high affinity. The search of additional external binding sites for fatty acids, distinct from the calyx, was also carried out for palmitic acid. Two external sites with a lower affinity were identified as secondary sites, one consisting in a hydrophobic cavity allowing two distinct binding modes for the fatty acid, and the other corresponding to a surface crevice close to the protein α-helix. The overall results provide a comprehensive picture of the dynamical behavior of βLG in complex with fatty acids, and elucidate the structural basis of the binding of these physiological ligands.

  12. The role of outer surface/inner bulk Brønsted acidic sites in the adsorption of a large basic molecule (simazine) on H-Y zeolite.

    PubMed

    Sannino, Filomena; Pansini, Michele; Marocco, Antonello; Bonelli, Barbara; Garrone, Edoardo; Esposito, Serena

    2015-11-21

    The simple means adopted for investigating H-Y zeolite acidity in water is the pH-dependence of the amount of a basic molecule adsorbed under isochoric conditions, a technique capable of yielding, under equilibrium conditions, an estimate of the pKa value of the involved acidic centres: the behaviour with temperature of adsorbed amounts yields instead some information on thermodynamics. Simazine (Sim, 2-chloro-4,6-bis(ethylamino)-s-triazine) was chosen as an adsorbate because its transverse dimension (7.5 Å) is close to the opening of the supercage in the faujasite structure of H-Y (7.4 Å). In short term measurements, Sim adsorption at 25 °C occurs only at the outer surface of H-Y particles. Two types of mildly acidic centres are present (with pKaca. 7 and ca. 8, respectively) and no strong one is observed. Previous adsorption of ammonia from the gas phase discriminates between the two. The former survives, and shows features common with the silanols of amorphous silica. The latter is suppressed: because of this and other features distinguishing this site from silanol species (e.g. the formation of dimeric Sim2H(+) species, favoured by coverage and unfavoured by temperatures of adsorption higher than ambient temperature) a candidate is an Al based site. We propose a Lewis centre coordinating a water molecule, exhibiting acidic properties. This acidic water molecule can be replaced by the stronger base ammonia, also depleting inner strong Brønsted sites. A mechanism for the generation of the two sites from surface Brønsted species is proposed. Long term adsorption measurements at 25 °C already show the onset of the interaction with inner strongly acidic Brønsted sites: because of its size, activation is required for Sim to pass the supercage openings and reach inner acidic sites. When adsorption is run at 40-50 °C, uptake is much larger and increases with temperature. Isochoric measurements suggest a pKa value of ca. 3 compatible with its marked acidic

  13. Amino acid sequence homology between Piv, an essential protein in site-specific DNA inversion in Moraxella lacunata, and transposases of an unusual family of insertion elements.

    PubMed Central

    Lenich, A G; Glasgow, A C

    1994-01-01

    Deletion analysis of the subcloned DNA inversion region of Moraxella lacunata indicates that Piv is the only M. lacunata-encoded factor required for site-specific inversion of the tfpQ/tfpI pilin segment. The predicted amino acid sequence of Piv shows significant homology solely with the transposases/integrases of a family of insertion sequence elements, suggesting that Piv is a novel site-specific recombinase. Images PMID:8021196

  14. Toughening of poly(lactic acid) without sacrificing stiffness and strength by melt-blending with polyamide 11 and selective localization of halloysite nanotubes

    NASA Astrophysics Data System (ADS)

    Rashmi, Baralu Jagannatha; Prashantha, Kalappa; Lacrampe, Marie-France; Krawczak, Patricia

    2016-03-01

    This paper aims at improving the mechanical behavior of biobased brittle amorphous polylactide (PLA) by extrusion melt-blending with biobased semi-crystalline polyamide 11 (PA 11) and addition of natural halloysite nanotubes (HNT). The structure and properties of PLA/PA11/HNT blends were studied in terms of morphological, thermal and mechanical properties. The morphological analysis of the PLA/PA11/HNT blends shows a strong interface between the two polymeric phases due to hydrogen bonding, and the migration of HNTs towards PA 11 phase inducing their selective localization in one of the polymeric phases of the blend. A "salami-like" structure is formed revealing a HNTs-rich tubular-like (fibrillar) PA11 phase. Moreover, HNTs localized in the dispersed phase acts as nucleating agents for PA11. Blending PLA (80 wt.%) and PA11 (20 wt.%) increases PLA ductility (elongation at break, ɛr, is multiplied by more than 20), however at the slight expense of strength and stiffness. Further addition of HNTs (2 wt.%) further increases ductility (ɛr reaches 155 %, i.e. it is multiplied by more than 40) whereas tensile strength and modulus of PLA are unchanged and impact strength is more than doubled. The toughening mechanism is discussed based on the combined effect of resistance to crack propagation and nanotubes load bearing capacity due to the existence of the fibrillar structure. Thus, blending brittle PLA with PA11 and HNT nanotubes results in tailor-made PLA-based compounds with enhanced ductility without sacrificing stiffness and strength.

  15. Acid-resistant calcium silicate-based composite implants with high-strength as load-bearing bone graft substitutes and fracture fixation devices.

    PubMed

    Wei, Chung-Kai; Ding, Shinn-Jyh

    2016-09-01

    To achieve the excellent mechanical properties of biodegradable materials used for cortical bone graft substitutes and fracture fixation devices remains a challenge. To this end, the biomimetic calcium silicate/gelatin/chitosan oligosaccharide composite implants were developed, with an aim of achieving high strength, controlled degradation, and superior osteogenic activity. The work focused on the effect of gelatin on mechanical properties of the composites under four different kinds of mechanical stresses including compression, tensile, bending, and impact. The evaluation of in vitro degradability and fatigue at two simulated body fluid (SBF) of pH 7.4 and 5.0 was also performed, in which the pH 5.0 condition simulated clinical conditions caused by bacterial induced local metabolic acidosis or tissue inflammation. In addition, human mesenchymal stem cells (hMSCs) were sued to examine osteogenic activity. Experimental results showed that the appropriate amount of gelatin positively contributed to failure enhancement in compressive and impact modes. The 10wt% gelatin-containing composite exhibits the maximum value of the compressive strength (166.1MPa), which is within the reported compressive strength for cortical bone. The stability of the bone implants was apparently affected by the in vitro fatigue, but not by the initial pH environments (7.4 or 5.0). The gelatin not only greatly enhanced the degradation of the composite when soaked in the dynamic SBF solution, but effectively promoted attachment, proliferation, differentiation, and formation of mineralization of hMSCs. The 10wt%-gelatin composite with high initial strength may be a potential implant candidate for cortical bone repair and fracture fixation applications. PMID:27254281

  16. Analysis of diurnal variability of atmospheric halocarbons and CFC replacements to imply emission strength and sources at an urban site of Lukang in central Taiwan

    NASA Astrophysics Data System (ADS)

    Lee, Bing-Sun; Chiou, Chung-Biau; Lin, Chung-Yi

    2014-12-01

    Hourly atmospheric measurements of halocarbons and chlorofluorocarbon (CFC) replacements were conducted at an urban site of Lukang, Changhua, in central Taiwan from May to August, 2013. The temporal distribution of different groups of halocarbons in the Lukang urban atmosphere, including chlorofluorocarbons (CFCs), Chlorodifluoromethane (HCFC-22), Bromochlorodifluoromethane (Halon-1211), and other chlorinated compounds, is presented and discussed. The concentrations (mixing ratios) of HCFC-22, Dichlorodifluoromethane (CFC-12), Halon-1211, Trichlorofluoromethane (CFC-11), Dichloromethane (CH2Cl2), and Trichloroethylene (TCE) were enhanced with respect to the local background levels; the atmospheric mixing ratio of carbon tetrachloride (CCl4) was slightly higher than its local background level; on the other hand, 1,1,2-Trichlorotrifluoroethane (CFC-113) was relatively uniform and not very different from background atmospheric level in non-urban areas. Among these compounds, HCFC-22, Halon-1211 and the halogenated compounds, CH2Cl2 and TCE, used as solvents were strongly enhanced. The average mixing ratio of Halon-1211 was higher than the local background of ∼4.5 ppt by ∼60% although Halon-1211 production had been phased out by 1996. Hourly average mixing ratios of halocarbons (HCFC-22, CFC-12, Halon-1211, CFC-11, CH2Cl2, and TCE) illustrated a distinct diurnal cycle characterized with a pattern of elevated mixing ratio and large mixing ratio variability amplitude at night relative to that in daytime. Although emission sources of these halocarbons were complex, hourly average mixing ratios for most of these high variability halocarbons peaked at ∼5:00 AM when the hourly average wind speed reached the minimum value of the day; by contrast, the hourly average mixing ratio of CO peaked at ∼8:30 AM when the ambient atmospheric wind condition was strongly influenced by sea breezes during the traffic rush hours. This phenomenon revealed that meteorological factors

  17. Site-directed mutagenesis of HgcA and HgcB reveals amino acid residues important for mercury methylation.

    PubMed

    Smith, Steven D; Bridou, Romain; Johs, Alexander; Parks, Jerry M; Elias, Dwayne A; Hurt, Richard A; Brown, Steven D; Podar, Mircea; Wall, Judy D

    2015-05-01

    Methylmercury is a potent neurotoxin that is produced by anaerobic microorganisms from inorganic mercury by a recently discovered pathway. A two-gene cluster, consisting of hgcA and hgcB, encodes two of the proteins essential for this activity. hgcA encodes a corrinoid protein with a strictly conserved cysteine proposed to be the ligand for cobalt in the corrinoid cofactor, whereas hgcB encodes a ferredoxin-like protein thought to be an electron donor to HgcA. Deletion of either gene eliminates mercury methylation by the methylator Desulfovibrio desulfuricans ND132. Here, site-directed mutants of HgcA and HgcB were constructed to determine amino acid residues essential for mercury methylation. Mutations of the strictly conserved residue Cys93 in HgcA, the proposed ligand for the corrinoid cobalt, to Ala or Thr completely abolished the methylation capacity, but a His substitution produced measurable methylmercury. Mutations of conserved amino acids near Cys93 had various impacts on the methylation capacity but showed that the structure of the putative "cap helix" region harboring Cys93 is crucial for methylation function. In the ferredoxin-like protein HgcB, only one of two conserved cysteines found at the C terminus was necessary for methylation, but either cysteine sufficed. An additional, strictly conserved cysteine, Cys73, was also determined to be essential for methylation. This study supports the previously predicted importance of Cys93 in HgcA for methylation of mercury and reveals additional residues in HgcA and HgcB that facilitate the production of this neurotoxin.

  18. Site-Directed Mutagenesis of HgcA and HgcB Reveals Amino Acid Residues Important for Mercury Methylation

    PubMed Central

    Smith, Steven D.; Bridou, Romain; Johs, Alexander; Parks, Jerry M.; Elias, Dwayne A.; Hurt, Richard A.; Brown, Steven D.; Podar, Mircea

    2015-01-01

    Methylmercury is a potent neurotoxin that is produced by anaerobic microorganisms from inorganic mercury by a recently discovered pathway. A two-gene cluster, consisting of hgcA and hgcB, encodes two of the proteins essential for this activity. hgcA encodes a corrinoid protein with a strictly conserved cysteine proposed to be the ligand for cobalt in the corrinoid cofactor, whereas hgcB encodes a ferredoxin-like protein thought to be an electron donor to HgcA. Deletion of either gene eliminates mercury methylation by the methylator Desulfovibrio desulfuricans ND132. Here, site-directed mutants of HgcA and HgcB were constructed to determine amino acid residues essential for mercury methylation. Mutations of the strictly conserved residue Cys93 in HgcA, the proposed ligand for the corrinoid cobalt, to Ala or Thr completely abolished the methylation capacity, but a His substitution produced measurable methylmercury. Mutations of conserved amino acids near Cys93 had various impacts on the methylation capacity but showed that the structure of the putative “cap helix” region harboring Cys93 is crucial for methylation function. In the ferredoxin-like protein HgcB, only one of two conserved cysteines found at the C terminus was necessary for methylation, but either cysteine sufficed. An additional, strictly conserved cysteine, Cys73, was also determined to be essential for methylation. This study supports the previously predicted importance of Cys93 in HgcA for methylation of mercury and reveals additional residues in HgcA and HgcB that facilitate the production of this neurotoxin. PMID:25724962

  19. Site-Directed Mutagenesis of HgcA and HgcB Reveals Amino Acid Residues Important for Mercury Methylation

    DOE PAGES

    Smith, Steven D.; Bridou, Romain; Johs, Alexander; Parks, Jerry M.; Elias, Dwayne A.; Hurt, Richard A.; Brown, Steven D.; Podar, Mircea; Wall, Judy D.

    2015-02-27

    Methylmercury is a potent neurotoxin that is produced by anaerobic microorganisms from inorganic mercury by a recently discovered pathway. A two-gene cluster, consisting of hgcA and hgcB, encodes two of the proteins essential for this activity. hgcA encodes a corrinoid protein with a strictly conserved cysteine proposed to be the ligand for cobalt in the corrinoid cofactor, whereas hgcB encodes a ferredoxin-like protein thought to be an electron donor to HgcA. Deletion of either gene eliminates mercury methylation by the methylator Desulfovibrio desulfuricans ND132. Here, site-directed mutants of HgcA and HgcB were constructed to determine amino acid residues essential formore » mercury methylation. Mutations of the strictly conserved residue Cys93 in HgcA, the proposed ligand for the corrinoid cobalt, to Ala or Thr completely abolished the methylation capacity, but a His substitution produced measurable methylmercury. Mutations of conserved amino acids near Cys93 had various impacts on the methylation capacity but showed that the structure of the putative “cap helix” region harboring Cys93 is crucial for methylation function. In the ferredoxin-like protein HgcB, only one of two conserved cysteines found at the C terminus was necessary for methylation, but either cysteine sufficed. An additional, strictly conserved cysteine, Cys73, was also determined to be essential for methylation. Ultimately, this study supports the previously predicted importance of Cys93 in HgcA for methylation of mercury and reveals additional residues in HgcA and HgcB that facilitate the production of this neurotoxin.« less

  20. The linoleic acid derivative DCP-LA selectively activates PKC-epsilon, possibly binding to the phosphatidylserine binding site.

    PubMed

    Kanno, Takeshi; Yamamoto, Hideyuki; Yaguchi, Takahiro; Hi, Rika; Mukasa, Takeshi; Fujikawa, Hirokazu; Nagata, Tetsu; Yamamoto, Satoshi; Tanaka, Akito; Nishizaki, Tomoyuki

    2006-06-01

    This study examined the effect of 8-[2-(2-pentyl-cyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA), a newly synthesized linoleic acid derivative with cyclopropane rings instead of cis-double bonds, on protein kinase C (PKC) activity. In the in situ PKC assay with reverse-phase high-performance liquid chromatography, DCP-LA significantly activated PKC in PC-12 cells in a concentration-dependent (10 nM-100 microM) manner, with the maximal effect at 100 nM, and the DCP-LA effect was blocked by GF109203X, a PKC inhibitor, or a selective inhibitor peptide of the novel PKC isozyme PKC-epsilon. Furthermore, DCP-LA activated PKC in HEK-293 cells that was inhibited by the small, interfering RNA against PKC-epsilon. In the cell-free PKC assay, of the nine isozymes examined here, DCP-LA most strongly activated PKC-epsilon, with >7-fold potency over other PKC isozymes, in the absence of dioleoyl-phosphatidylserine and 1,2-dioleoyl-sn-glycerol; instead, the DCP-LA action was inhibited by dioleoyl-phosphatidylserine. DCP-LA also activated PKC-gamma, a conventional PKC, but to a much lesser extent compared with that for PKC-epsilon, by a mechanism distinct from PKC-epsilon activation. Thus, DCP-LA serves as a selective activator of PKC-epsilon, possibly by binding to the phosphatidylserine binding site on PKC-epsilon. These results may provide fresh insight into lipid signaling in PKC activation.

  1. Site-Directed Mutagenesis of HgcA and HgcB Reveals Amino Acid Residues Important for Mercury Methylation

    SciTech Connect

    Smith, Steven D.; Bridou, Romain; Johs, Alexander; Parks, Jerry M.; Elias, Dwayne A.; Hurt, Richard A.; Brown, Steven D.; Podar, Mircea; Wall, Judy D.

    2015-02-27

    Methylmercury is a potent neurotoxin that is produced by anaerobic microorganisms from inorganic mercury by a recently discovered pathway. A two-gene cluster, consisting of hgcA and hgcB, encodes two of the proteins essential for this activity. hgcA encodes a corrinoid protein with a strictly conserved cysteine proposed to be the ligand for cobalt in the corrinoid cofactor, whereas hgcB encodes a ferredoxin-like protein thought to be an electron donor to HgcA. Deletion of either gene eliminates mercury methylation by the methylator Desulfovibrio desulfuricans ND132. Here, site-directed mutants of HgcA and HgcB were constructed to determine amino acid residues essential for mercury methylation. Mutations of the strictly conserved residue Cys93 in HgcA, the proposed ligand for the corrinoid cobalt, to Ala or Thr completely abolished the methylation capacity, but a His substitution produced measurable methylmercury. Mutations of conserved amino acids near Cys93 had various impacts on the methylation capacity but showed that the structure of the putative “cap helix” region harboring Cys93 is crucial for methylation function. In the ferredoxin-like protein HgcB, only one of two conserved cysteines found at the C terminus was necessary for methylation, but either cysteine sufficed. An additional, strictly conserved cysteine, Cys73, was also determined to be essential for methylation. Ultimately, this study supports the previously predicted importance of Cys93 in HgcA for methylation of mercury and reveals additional residues in HgcA and HgcB that facilitate the production of this neurotoxin.

  2. In Silico Structure Prediction of Human Fatty Acid Synthase–Dehydratase: A Plausible Model for Understanding Active Site Interactions

    PubMed Central

    John, Arun; Umashankar, Vetrivel; Samdani, A.; Sangeetha, Manoharan; Krishnakumar, Subramanian; Deepa, Perinkulam Ravi

    2016-01-01

    Fatty acid synthase (FASN, UniProt ID: P49327) is a multienzyme dimer complex that plays a critical role in lipogenesis. Consequently, this lipogenic enzyme has gained tremendous biomedical importance. The role of FASN and its inhibition is being extensively researched in several clinical conditions, such as cancers, obesity, and diabetes. X-ray crystallographic structures of some of its domains, such as β-ketoacyl synthase, acetyl transacylase, malonyl transacylase, enoyl reductase, β-ketoacyl reductase, and thioesterase, (TE) are already reported. Here, we have attempted an in silico elucidation of the uncrystallized dehydratase (DH) catalytic domain of human FASN. This theoretical model for DH domain was predicted using comparative modeling methods. Different stand-alone tools and servers were used to validate and check the reliability of the predicted models, which suggested it to be a highly plausible model. The stereochemical analysis showed 92.0% residues in favorable region of Ramachandran plot. The initial physiological substrate β-hydroxybutyryl group was docked into active site of DH domain using Glide. The molecular dynamics simulations carried out for 20 ns in apo and holo states indicated the stability and accuracy of the predicted structure in solvated condition. The predicted model provided useful biochemical insights into the substrate–active site binding mechanisms. This model was then used for identifying potential FASN inhibitors using high-throughput virtual screening of the National Cancer Institute database of chemical ligands. The inhibitory efficacy of the top hit ligands was validated by performing molecular dynamics simulation for 20 ns, where in the ligand NSC71039 exhibited good enzyme inhibition characteristics and exhibited dose-dependent anticancer cytotoxicity in retinoblastoma cancer cells in vitro. PMID:27559295

  3. In Silico Structure Prediction of Human Fatty Acid Synthase-Dehydratase: A Plausible Model for Understanding Active Site Interactions.

    PubMed

    John, Arun; Umashankar, Vetrivel; Samdani, A; Sangeetha, Manoharan; Krishnakumar, Subramanian; Deepa, Perinkulam Ravi

    2016-01-01

    Fatty acid synthase (FASN, UniProt ID: P49327) is a multienzyme dimer complex that plays a critical role in lipogenesis. Consequently, this lipogenic enzyme has gained tremendous biomedical importance. The role of FASN and its inhibition is being extensively researched in several clinical conditions, such as cancers, obesity, and diabetes. X-ray crystallographic structures of some of its domains, such as β-ketoacyl synthase, acetyl transacylase, malonyl transacylase, enoyl reductase, β-ketoacyl reductase, and thioesterase, (TE) are already reported. Here, we have attempted an in silico elucidation of the uncrystallized dehydratase (DH) catalytic domain of human FASN. This theoretical model for DH domain was predicted using comparative modeling methods. Different stand-alone tools and servers were used to validate and check the reliability of the predicted models, which suggested it to be a highly plausible model. The stereochemical analysis showed 92.0% residues in favorable region of Ramachandran plot. The initial physiological substrate β-hydroxybutyryl group was docked into active site of DH domain using Glide. The molecular dynamics simulations carried out for 20 ns in apo and holo states indicated the stability and accuracy of the predicted structure in solvated condition. The predicted model provided useful biochemical insights into the substrate-active site binding mechanisms. This model was then used for identifying potential FASN inhibitors using high-throughput virtual screening of the National Cancer Institute database of chemical ligands. The inhibitory efficacy of the top hit ligands was validated by performing molecular dynamics simulation for 20 ns, where in the ligand NSC71039 exhibited good enzyme inhibition characteristics and exhibited dose-dependent anticancer cytotoxicity in retinoblastoma cancer cells in vitro. PMID:27559295

  4. Effect of different soil layers on porewater to remediate acidic surface environment at a close mine site.

    PubMed

    Salinas Villafane, Omar R; Igarashi, Toshifumi; Harada, Shusaku; Kurosawa, Mitsuru; Takase, Toshio

    2012-12-01

    This paper describes the chemistry of porewater when constructing different soil layers on acidic weathered rock of a closed mine to remediate the surface environment. Three cases were set on a flat surface of the site, all under different layer systems. Case 1 was only composed of weathered rocks. A top neutralization layer was constructed on the weathered rocks in case 2, whereas both an upper low-permeable and middle neutralization layers were constructed on the weathered rocks in case 3. The low-permeable layer of 30 cm thick consists of clay, and the neutralization layer of 30 cm thick consists of the mixture of the weathered rock and calcium carbonate as a neutralizer. Porewater sampling systems and soil sensors to measure temperature, water content, and electrical conductivity were set at different depths. In case 1, steadily high concentrations of heavy metals were observed regardless of the depth, and the pH ranged from 2 to 4. In cases 2 and 3, a dramatic decrease in concentrations of heavy metals was observed, even below the neutralization layer. For both cases, pH values were circumneutral. There were no significant seasonable changes in heavy metals concentrations and pH of porewater by considering the temperature and precipitation. In addition, the water content of the layers in case 3 fluctuated more mildly than that in cases 1 and 2, indicating that the low-permeable layer reduced the rate of infiltration. Therefore, a significant reduction in the load of heavy metals released from the site can be achieved by both implementing neutralization and low-permeable layers.

  5. Creation of a thermostable NADP⁺-dependent D-amino acid dehydrogenase from Ureibacillus thermosphaericus strain A1 meso-diaminopimelate dehydrogenase by site-directed mutagenesis.

    PubMed

    Akita, Hironaga; Doi, Katsumi; Kawarabayasi, Yutaka; Ohshima, Toshihisa

    2012-09-01

    A thermostable, NADP(+)-dependent D: -amino acid dehydrogenase (DAADH) was created from the meso-diaminopimelate dehydrogenase of Ureibacillus thermosphaericus strain A1 by introducing five point mutations into amino acid residues located in the active site. The recombinant protein, expressed in Escherichia coli, was purified to homogeneity using a two-step separation procedure and then characterized. In the presence of NADP(+), the protein catalyzed the oxidative deamination of several D: -amino acids, including D: -cyclohexylalanine, D: -isoleucine and D: -2-aminooctanoate, but not meso-diaminopimelate, confirming the creation of a NADP(+)-dependent DAADH. For the reverse reaction, the corresponding 2-oxo acids were aminated in the presence of NADPH and ammonia. In addition, the D: -amino acid dehydrogenase showed no loss of activity at 65 °C, indicating the mutant enzyme was more thermostable than its parental meso-diaminopimelate dehydrogenase.

  6. Chemical repair of base lesions, AP-sites, and strand breaks on plasmid DNA in dilute aqueous solution by ascorbic acid

    SciTech Connect

    Hata, Kuniki; Urushibara, Ayumi; Yamashita, Shinichi; Shikazono, Naoya; Yokoya, Akinari; Katsumura, Yosuke

    2013-05-03

    Highlights: •We report a novel mechanism of radiation protection of DNA by chemical activity of ascorbic acid. •The “chemical repair” of DNA damage was revealed using biochemical assay and chemical kinetics analysis. •We found that ascorbic acid significantly repairs precursors of nucleobase lesions and abasic sites. •However, ascorbic acid seldom repairs precursors of DNA-strand breaks. -- Abstract: We quantified the damage yields produced in plasmid DNA by γ-irradiation in the presence of low concentrations (10–100 μM) of ascorbic acid, which is a major antioxidant in living systems, to clarify whether it chemically repairs radiation damage in DNA. The yield of DNA single strand breaks induced by irradiation was analyzed with agarose gel electrophoresis as conformational changes in closed circular plasmids. Base lesions and abasic sites were also observed as additional conformational changes by treating irradiated samples with glycosylase proteins. By comparing the suppression efficiencies to the induction of each DNA lesion, in addition to scavenging of the OH radicals derived from water radiolysis, it was found that ascorbic acid promotes the chemical repair of precursors of AP-sites and base lesions more effectively than those of single strand breaks. We estimated the efficiency of the chemical repair of each lesion using a kinetic model. Approximately 50–60% of base lesions and AP-sites were repaired by 10 μM ascorbic acid, although strand breaks were largely unrepaired by ascorbic acid at low concentrations. The methods in this study will provide a route to understanding the mechanistic aspects of antioxidant activity in living systems.

  7. Key Amino Acid Residues of Ankyrin-Sensitive Phosphatidylethanolamine/Phosphatidylcholine-Lipid Binding Site of βI-Spectrin

    PubMed Central

    Wolny, Marcin; Grzybek, Michał; Bok, Ewa; Chorzalska, Anna; Lenoir, Marc; Czogalla, Aleksander; Adamczyk, Klaudia; Kolondra, Adam; Diakowski, Witold; Overduin, Michael; Sikorski, Aleksander F.

    2011-01-01

    It was shown previously that an ankyrin-sensitive, phosphatidylethanolamine/phosphatidylcholine (PE/PC) binding site maps to the N-terminal part of the ankyrin-binding domain of β-spectrin (ankBDn). Here we have identified the amino acid residues within this domain which are responsible for recognizing monolayers and bilayers composed of PE/PC mixtures. In vitro binding studies revealed that a quadruple mutant with substituted hydrophobic residues W1771, L1775, M1778 and W1779 not only failed to effectively bind PE/PC, but its residual PE/PC-binding activity was insensitive to inhibition with ankyrin. Structure prediction and analysis, supported by in vitro experiments, suggests that “opening” of the coiled-coil structure underlies the mechanism of this interaction. Experiments on red blood cells and HeLa cells supported the conclusions derived from the model and in vitro lipid-protein interaction results, and showed the potential physiological role of this binding. We postulate that direct interactions between spectrin ankBDn and PE-rich domains play an important role in stabilizing the structure of the spectrin-based membrane skeleton. PMID:21738695

  8. Efficient Catalytic Ozonation over Reduced Graphene Oxide for p-Hydroxylbenzoic Acid (PHBA) Destruction: Active Site and Mechanism.

    PubMed

    Wang, Yuxian; Xie, Yongbing; Sun, Hongqi; Xiao, Jiadong; Cao, Hongbin; Wang, Shaobin

    2016-04-20

    Nanocarbons have been demonstrated as promising environmentally benign catalysts for advanced oxidation processes (AOPs) upgrading metal-based materials. In this study, reduced graphene oxide (rGO) with a low level of structural defects was synthesized via a scalable method for catalytic ozonation of p-hydroxylbenzoic acid (PHBA). Metal-free rGO materials were found to exhibit a superior activity in activating ozone for catalytic oxidation of organic phenolics. The electron-rich carbonyl groups were identified as the active sites for the catalytic reaction. Electron spin resonance (ESR) and radical competition tests revealed that superoxide radical ((•)O2(-)) and singlet oxygen ((1)O2) were the reactive oxygen species (ROS) for PHBA degradation. The intermediates and the degradation pathways were illustrated from mass spectroscopy. It was interesting to observe that addition of NaCl could enhance both ozonation and catalytic ozonation efficiencies and make ·O2(-) as the dominant ROS. Stability of the catalysts was also evaluated by the successive tests. Loss of specific surface area and changes in the surface chemistry were suggested to be responsible for catalyst deactivation. PMID:27007603

  9. Site-specific influence of polyunsaturated fatty acids on atherosclerosis in immune incompetent LDL receptor deficient mice.

    PubMed

    Reardon, Catherine A; Blachowicz, Lydia; Gupta, Gaorav; Lukens, John; Nissenbaum, Michael; Getz, Godfrey S

    2006-08-01

    Polyunsaturated fatty acids (PUFA) are thought to influence plasma lipid levels, atherosclerosis, and the immune system. In this study, we fed male LDL receptor deficient (LDLR(-/-)) mice and immune incompetent LDLR(-/-) RAG2(-/-) mice diets containing predominantly saturated fats (milk fat) or PUFA (safflower oil) to determine if the response to diet was influenced by immune status. Relative to milk fat diet, plasma lipid and VLDL levels in both the LDLR(-/-) and LDLR(-/-) RAG2(-/-) mice fed safflower oil diet were lower, suggesting that the primary effect of PUFA on plasma lipids was not due to its inhibition of the immune system. Neither diet nor immune status influenced hepatic triglyceride production and post-heparin lipase activity, suggesting that the differences in triglyceride levels are due to differences in rates of catabolism of triglyceride-rich lipoproteins. While both diets promoted atherogenesis, both aortic root and innominate artery atherosclerosis in LDLR(-/-) mice was less in safflower oil fed animals. In contrast, a site-specific effect of PUFA was observed in the immune incompetent LDLR(-/-) RAG2(-/-). In these mice, aortic root atherosclerosis, but not innominate artery atherosclerosis, was less in PUFA fed animal. These results suggest that PUFA and the immune system may influence innominate artery atherosclerosis by some overlapping mechanisms.

  10. "Opening" the ferritin pore for iron release by mutation of conserved amino acids at interhelix and loop sites.

    PubMed

    Jin, W; Takagi, H; Pancorbo, B; Theil, E C

    2001-06-26

    Ferritin concentrates, stores, and detoxifies iron in most organisms. The iron is a solid, ferric oxide mineral (< or =4500 Fe) inside the protein shell. Eight pores are formed by subunit trimers of the 24 subunit protein. A role for the protein in controlling reduction and dissolution of the iron mineral was suggested in preliminary experiments [Takagi et al. (1998) J. Biol. Chem. 273, 18685-18688] with a proline/leucine substitution near the pore. Localized pore disorder in frog L134P crystals coincided with enhanced iron exit, triggered by reduction. In this report, nine additional substitutions of conserved amino acids near L134 were studied for effects on iron release. Alterations of a conserved hydrophobic pair, a conserved ion pair, and a loop at the ferritin pores all increased iron exit (3-30-fold). Protein assembly was unchanged, except for a slight decrease in volume (measured by gel filtration); ferroxidase activity was still in the millisecond range, but a small decrease indicates slight alteration of the channel from the pore to the oxidation site. The sensitivity of reductive iron exit rates to changes in conserved residues near the ferritin pores, associated with localized unfolding, suggests that the structure around the ferritin pores is a target for regulated protein unfolding and iron release.

  11. Acid-Sensitive Sheddable PEGylated PLGA Nanoparticles Increase the Delivery of TNF-α siRNA in Chronic Inflammation Sites.

    PubMed

    Aldayel, Abdulaziz M; Naguib, Youssef W; O'Mary, Hannah L; Li, Xu; Niu, Mengmeng; Ruwona, Tinashe B; Cui, Zhengrong

    2016-01-01

    There has been growing interest in utilizing small interfering RNA (siRNA) specific to pro-inflammatory cytokines, such as tumor necrosis factor-α ( TNF-α), in chronic inflammation therapy. However, delivery systems that can increase the distribution of the siRNA in chronic inflammation sites after intravenous administration are needed. Herein we report that innovative functionalization of the surface of siRNA-incorporated poly (lactic-co-glycolic) acid (PLGA) nanoparticles significantly increases the delivery of the siRNA in the chronic inflammation sites in a mouse model. The TNF-α siRNA incorporated PLGA nanoparticles were prepared by the standard double emulsion method, but using stearoyl-hydrazone-polyethylene glycol 2000, a unique acid-sensitive surface active agent, as the emulsifying agent, which renders (i) the nanoparticles PEGylated and (ii) the PEGylation sheddable in low pH environment such as that in chronic inflammation sites. In a mouse model of lipopolysaccharide-induced chronic inflammation, the acid-sensitive sheddable PEGylated PLGA nanoparticles showed significantly higher accumulation or distribution in chronic inflammation sites than PLGA nanoparticles prepared with an acid-insensitive emulsifying agent (i.e., stearoyl-amide-polyethylene glycol 2000) and significantly increased the distribution of the TNF-α siRNA incorporated into the nanoparticles in inflamed mouse foot. PMID:27434685

  12. Bond strength of composite to dentin: effect of acid etching and laser irradiation through an uncured self-etch adhesive system

    NASA Astrophysics Data System (ADS)

    Castro, F. L. A.; Carvalho, J. G.; Andrade, M. F.; Saad, J. R. C.; Hebling, J.; Lizarelli, R. F. Z.

    2014-08-01

    This study evaluated the effect on micro-tensile bond strength (µ-TBS) of laser irradiation of etched/unetched dentin through an uncured self-etching adhesive. Dentinal surfaces were treated with Clearfil SE Bond Adhesive (CSE) either according to the manufacturer’s instructions (CSE) or without applying the primer (CSE/NP). The dentin was irradiated through the uncured adhesive, using an Nd:YAG laser at 0.75 or 1 W power settings. The adhesive was cured, composite crowns were built up, and the teeth were sectioned into beams (0.49 mm2) to be stressed under tension. Data were analyzed using one-way ANOVA and Tukey statistics (α = 5%). Dentin of the fractured specimens and the interfaces of untested beams were observed under scanning electron microscopy (SEM). The results showed that non-etched irradiated surfaces presented higher µ-TBS than etched and irradiated surfaces (p < 0.05). Laser irradiation alone did not lead to differences in µ-TBS (p > 0.05). SEM showed solidification globules on the surfaces of the specimens. The interfaces were similar on irradiated and non-irradiated surfaces. Laser irradiation of dentin through the uncured adhesive did not lead to higher µ-TBS when compared to the suggested manufacturer’s technique. However, this treatment brought benefits when performed on unetched dentin, since bond strengths were higher when compared to etched dentin.

  13. Effects of pH, dissolved oxygen, and ionic strength on the survival of Escherichia coli O157:H7 in organic acid solutions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ability of Escherichia coli O157:H7 to survive in acidified vegetable products is of concern because of previously documented outbreaks associated with fruit juices. A study was conducted to determine the survival of E. coli O157:H7 in organic acids at pH values typical of acidified vegetable pr...

  14. Mapping general anesthetic binding site(s) in human α1β3 γ-aminobutyric acid type A receptors with [³H]TDBzl-etomidate, a photoreactive etomidate analogue.

    PubMed

    Chiara, David C; Dostalova, Zuzana; Jayakar, Selwyn S; Zhou, Xiaojuan; Miller, Keith W; Cohen, Jonathan B

    2012-01-31

    The γ-aminobutyric acid type A receptor (GABA(A)R) is a target for general anesthetics of diverse chemical structures, which act as positive allosteric modulators at clinical doses. Previously, in a heterogeneous mixture of GABA(A)Rs purified from bovine brain, [³H]azietomidate photolabeling of αMet-236 and βMet-286 in the αM1 and βM3 transmembrane helices identified an etomidate binding site in the GABA(A)R transmembrane domain at the interface between the β and α subunits [Li, G. D., et.al. (2006) J. Neurosci. 26, 11599-11605]. To further define GABA(A)R etomidate binding sites, we now use [³H]TDBzl-etomidate, an aryl diazirine with broader amino acid side chain reactivity than azietomidate, to photolabel purified human FLAG-α1β3 GABA(A)Rs and more extensively identify photolabeled GABA(A)R amino acids. [³H]TDBzl-etomidate photolabeled in an etomidate-inhibitable manner β3Val-290, in the β3M3 transmembrane helix, as well as α1Met-236 in α1M1, a residue photolabeled by [³H]azietomidate, while no photolabeling of amino acids in the αM2 and βM2 helices that also border the etomidate binding site was detected. The location of these photolabeled amino acids in GABA(A)R homology models derived from the recently determined structures of prokaryote (GLIC) or invertebrate (GluCl) homologues and the results of computational docking studies predict the orientation of [³H]TDBzl-etomidate bound in that site and the other amino acids contributing to this GABA(A)R intersubunit etomidate binding site. Etomidate-inhibitable photolabeling of β3Met-227 in βM1 by [³H]TDBzl-etomidate and [³H]azietomidate also provides evidence of a homologous etomidate binding site at the β3-β3 subunit interface in the α1β3 GABA(A)R.

  15. Enhancement of the catalytic activity of ferulic acid decarboxylase from Enterobacter sp. Px6-4 through random and site-directed mutagenesis.

    PubMed

    Lee, Hyunji; Park, Jiyoung; Jung, Chaewon; Han, Dongfei; Seo, Jiyoung; Ahn, Joong-Hoon; Chong, Youhoon; Hur, Hor-Gil

    2015-11-01

    The enzyme ferulic acid decarboxylase (FADase) from Enterobacter sp. Px6-4 catalyzes the decarboxylation reaction of lignin monomers and phenolic compounds such as p-coumaric acid, caffeic acid, and ferulic acid into their corresponding 4-vinyl derivatives, that is, 4-vinylphenol, 4-vinylcatechol, and 4-vinylguaiacol, respectively. Among various ferulic acid decarboxylase enzymes, we chose the FADase from Enterobacter sp. Px6-4, whose crystal structure is known, and produced mutants to enhance its catalytic activity by random and site-directed mutagenesis. After three rounds of sequential mutations, FADase(F95L/D112N/V151I) showed approximately 34-fold higher catalytic activity than wild-type for the production of 4-vinylguaiacol from ferulic acid. Docking analyses suggested that the increased activity of FADase(F95L/D112N/V151I) could be due to formation of compact active site compared with that of the wild-type FADase. Considering the amount of phenolic compounds such as lignin monomers in the biomass components, successfully bioengineered FADase(F95L/D112N/V151I) from Enterobacter sp. Px6-4 could provide an ecofriendly biocatalytic tool for producing diverse styrene derivatives from biomass.

  16. Enhancement of the catalytic activity of ferulic acid decarboxylase from Enterobacter sp. Px6-4 through random and site-directed mutagenesis.

    PubMed

    Lee, Hyunji; Park, Jiyoung; Jung, Chaewon; Han, Dongfei; Seo, Jiyoung; Ahn, Joong-Hoon; Chong, Youhoon; Hur, Hor-Gil

    2015-11-01

    The enzyme ferulic acid decarboxylase (FADase) from Enterobacter sp. Px6-4 catalyzes the decarboxylation reaction of lignin monomers and phenolic compounds such as p-coumaric acid, caffeic acid, and ferulic acid into their corresponding 4-vinyl derivatives, that is, 4-vinylphenol, 4-vinylcatechol, and 4-vinylguaiacol, respectively. Among various ferulic acid decarboxylase enzymes, we chose the FADase from Enterobacter sp. Px6-4, whose crystal structure is known, and produced mutants to enhance its catalytic activity by random and site-directed mutagenesis. After three rounds of sequential mutations, FADase(F95L/D112N/V151I) showed approximately 34-fold higher catalytic activity than wild-type for the production of 4-vinylguaiacol from ferulic acid. Docking analyses suggested that the increased activity of FADase(F95L/D112N/V151I) could be due to formation of compact active site compared with that of the wild-type FADase. Considering the amount of phenolic compounds such as lignin monomers in the biomass components, successfully bioengineered FADase(F95L/D112N/V151I) from Enterobacter sp. Px6-4 could provide an ecofriendly biocatalytic tool for producing diverse styrene derivatives from biomass. PMID:26059194

  17. D-aspartic acid supplementation combined with 28 days of heavy resistance training has no effect on body composition, muscle strength, and serum hormones associated with the hypothalamo-pituitary-gonadal axis in resistance-trained men.

    PubMed

    Willoughby, Darryn S; Leutholtz, Brian

    2013-10-01

    It was hypothesized that D-aspartic acid (D-ASP) supplementation would not increase endogenous testosterone levels or improve muscular performance associated with resistance training. Therefore, body composition, muscle strength, and serum hormone levels associated with the hypothalamo-pituitary-gonadal axis were studied after 28 days of resistance training and D-ASP supplementation. Resistance-trained men resistance trained 4 times/wk for 28 days while orally ingesting either 3 g of placebo or 3 g of D-ASP. Data were analyzed with 2 × 2 analysis of variance (P < .05). Before and after resistance training and supplementation, body composition and muscle strength, serum gonadal hormones, and serum D-ASP and d-aspartate oxidase (DDO) were determined. Body composition and muscle strength were significantly increased in both groups in response to resistance training (P < .05) but not different from one another (P > .05). Total and free testosterone, luteinizing hormone, gonadotropin-releasing hormone, and estradiol were unchanged with resistance training and D-ASP supplementation (P > .05). For serum D-ASP and DDO, D-ASP resulted in a slight increase compared with baseline levels (P > .05). For the D-ASP group, the levels of serum DDO were significantly increased compared with placebo (P < .05). The gonadal hormones were unaffected by 28 days of D-ASP supplementation and not associated with the observed increases in muscle strength and mass. Therefore, at the dose provided, D-ASP supplementation is ineffective in up-regulating the activity of the hypothalamo-pituitary-gonadal axis and has no anabolic or ergogenic effects in skeletal muscle.

  18. Active Site Dynamical Effects in the Hydrogen Transfer Rate-limiting Step in the Catalysis of Linoleic Acid by Soybean Lipoxygenase-1 (SLO-1): Primary and Secondary Isotope Contributions.

    PubMed

    Phatak, Prasad; Venderley, Jordan; Debrota, John; Li, Junjie; Iyengar, Srinivasan S

    2015-07-30

    Using ab initio molecular dynamics (AIMD) simulations that facilitate the treatment of rare events, we probe the active site participation in the rate-determining hydrogen transfer step in the catalytic oxidation of linoleic acid by soybean lipoxygenase-1 (SLO-1). The role of two different active site components is probed. (a) On the hydrogen atom acceptor side of the active site, the hydrogen bonding propensity between the acceptor side hydroxyl group, which is bound to the iron cofactor, and the backbone carboxyl group of isoleucine (residue number 839) is studied toward its role in promoting the hydrogen transfer event. Primary and secondary (H/D) isotope effects are also probed and a definite correlation with subtle secondary H/D isotope effects is found. With increasing average nuclear kinetic energy, the increase in transfer probability is enhanced due to the presence of the hydrogen bond between the backbone carbonyl of I839 and the acceptor oxygen. Further increase in average nuclear kinetic energy reduces the strength of this secondary hydrogen bond which leads to a deterioration in hydrogen transfer rates and finally embrances an Arrhenius-like behavior. (b) On the hydrogen atom donor side, the coupling between vibrational modes predominantly localized on the donor-side linoleic acid group and the reactive mode is probed. There appears to be a qualitative difference in the coupling between modes that belong to linoleic acid and the hydrogen transfer mode, for hydrogen and deuterium transfer. For example, the donor side secondary hydrogen atom is much more labile (by nearly a factor of 5) during deuterium transfer as compared to the case for hydrogen transfer. This appears to indicate a greater coupling between the modes belonging to the linoleic acid scaffold and the deuterium transfer mode and also provides a new rationalization for the abnormal (nonclassical) secondary isotope effect results obtained by Knapp, Rickert, and Klinman in J. Am. Chem. Soc

  19. Effects of folic acid on overall and site-specific cancer incidence during the randomised trials: meta-analyses of data on 50 000 individuals

    PubMed Central

    Vollset, Stein Emil; Clarke, Robert; Lewington, Sarah; Ebbing, Marta; Halsey, Jim; Lonn, Eva; Armitage, Jane; Manson, JoAnn E; Hankey, Graeme J; Spence, J David; Galan, Pilar; Bønaa, Kaare H; Jamison, Rex; Gaziano, J Michael; Guarino, Peter; Baron, John A; Logan, Richard FA; Giovannucci, Edward L; den Heijer, Martin; Ueland, Per M; Bennett, Derrick; Collins, Rory; Peto, Richard

    2013-01-01

    Summary Background Some countries fortify flour with folic acid to prevent neural tube defects but others do not, partly because of concerns about cancer risks. We aimed to assess the effects of folic acid supplementation on site-specific cancer rates in the randomised trials. Methods Meta-analyses of data on each individual in all placebo-controlled trials of folic acid for prevention of cardiovascular disease (10 trials, n=46,969) or colorectal adenoma (3 trials, n=2652) that recorded cancer incidence and recruited >500 participants. All trials were evenly randomised. Risk ratios (RRs) compare those allocated folic acid vs those allocated placebo, giving cancer incidence rate ratios (among those still free of cancer) during, but not after the scheduled treatment period. Findings During a weighted mean follow-up duration of 5.5 years, allocation to folic acid quadrupled plasma folate, but had no statistically significant effect on overall cancer incidence (1904 vs 1809 cancers, RR=1.06 [95%CI 0.99–1.13], p=0.10; trend with duration of treatment p=0.46). There was no significant heterogeneity between the results of individual trials (p=0.23), or between the cadiovascular prevention trials and the adenoma prevention trials (p=0.13). Moreover, there was no significant effect of folic acid supplementation on the incidence of cancer of the large intestine, prostate, lung, breast or any other specific site. Interpretation Folic acid supplementation does not substantially increase or decrease site-specific cancer incidence during the first 5 years of treatment. Funding British Heart Foundation, Medical Research Council, Cancer Research UK, Food Standards Agency. PMID:23352552

  20. Fatty acid-binding site environments of serum vitamin D-binding protein and albumin are different

    PubMed Central

    Swamy, Narasimha; Ray, Rahul

    2008-01-01

    Vitamin D-binding protein (DBP) and albumin (ALB) are abundant serum proteins and both possess high-affinity binding for saturated and unsaturated fatty acids. However, certain differences exist. We surmised that in cases where serum albumin level is low, DBP presumably can act as a transporter of fatty acids. To explore this possibility we synthesized several alkylating derivatives of 14C-palmitic acid to probe the fatty acid binding pockets of DBP and ALB. We observed that N-ethyl-5-phenylisooxazolium-3′-sulfonate-ester (WRK ester) of 14C-palmitic acid specifically labeled DBP; but p-nitrophenyl- and N-hydroxysuccinimidyl-esters failed to do so. However, p-nitrophenyl ester of 14C-palmitic acid specifically labeled bovine ALB, indicating that the micro-environment of the fatty acid-binding domains of DBP and ALB may be different; and DBP may not replace ALB as a transporter of fatty acids. PMID:18374965

  1. Identification of glutamic acid 78 as the active site nucleophile in Bacillus subtilis xylanase using electrospray tandem mass spectrometry.

    PubMed

    Miao, S; Ziser, L; Aebersold, R; Withers, S G

    1994-06-14

    A new mechanism-based inactivator of beta-1,4-xylanases, 2',4'-dinitrophenyl 2-deoxy-2-fluoro-beta-xylobioside, has been synthesized and used to trap the covalent intermediate formed during catalysis by Bacillus subtilis xylanase. Electrospray mass spectrometry confirmed the 1:1 stoichiometry of the incorporation of inactivator into the enzyme. Inactivation of xylanase followed the expected pseudo-first-order kinetic behavior, and kinetic parameters were determined. The intermediate trapped was relatively stable toward hydrolytic turnover (t1/2 = 350 min). However, turnover could be facilitated by transglycosylation following the addition of the acceptor benzyl thio-beta-xylobioside, thus demonstrating the catalytic competence of the trapped intermediate. Reactivation kinetic parameters for this process of kre = 0.03 min-1 and Kre = 46 mM were determined. The nucleophilic amino acid was identified as Glu78 by a tandem mass spectrometric technique which does not require the use of radiolabels. The peptic digest of the labeled enzyme was separated by high-performance liquid chromatography and the eluent fed into a tandem mass spectrometer via an electrospray ionization device. The labeled peptide was identified as one of m/z = 826 (doubly charged) which fragmented in the collision chamber between the mass analyzers with loss of the mass of a 2-fluoroxylobiosyl unit. Confirmation of the peptide identity was obtained both by tandem mass spectrometric sequencing and by Edman degradation of the purified peptide. Glu78 is completely conserved in all members of this xylanase family and indeed is shown to be located in the active site in the recently determined X-ray crystal structure.

  2. Application of Strength Diagnosis.

    ERIC Educational Resources Information Center

    Newton, Robert U.; Dugan, Eric

    2002-01-01

    Discusses the various strength qualities (maximum strength, high- and low-load speed strength, reactive strength, rate of force development, and skill performance), noting why a training program design based on strength diagnosis can lead to greater efficacy and better performance gains for the athlete. Examples of tests used to assess strength…

  3. Characterization of the N-Acetyl-5-neuraminic Acid-binding Site of the Extracytoplasmic Solute Receptor (SiaP) of Nontypeable Haemophilus influenzae Strain 2019

    SciTech Connect

    Johnston, Jason W.; Coussens, Nathan P.; Allen, Simon; Houtman, Jon C.D.; Turner, Keith H.; Zaleski, Anthony; Ramaswamy, S.; Gibson, Bradford W.; Apicella, Michael A.

    2012-11-14

    Nontypeable Haemophilus influenzae is an opportunistic human pathogen causing otitis media in children and chronic bronchitis and pneumonia in patients with chronic obstructive pulmonary disease. The outer membrane of nontypeable H. influenzae is dominated by lipooligosaccharides (LOS), many of which incorporate sialic acid as a terminal nonreducing sugar. Sialic acid has been demonstrated to be an important factor in the survival of the bacteria within the host environment. H. influenzae is incapable of synthesizing sialic acid and is dependent on scavenging free sialic acid from the host environment. To achieve this, H. influenzae utilizes a tripartite ATP-independent periplasmic transporter. In this study, we characterize the binding site of the extracytoplasmic solute receptor (SiaP) from nontypeable H. influenzae strain 2019. A crystal structure of N-acetyl-5-neuraminic acid (Neu5Ac)-bound SiaP was determined to 1.4 {angstrom} resolution. Thermodynamic characterization of Neu5Ac binding shows this interaction is enthalpically driven with a substantial unfavorable contribution from entropy. This is expected because the binding of SiaP to Neu5Ac is mediated by numerous hydrogen bonds and has several buried water molecules. Point mutations targeting specific amino acids were introduced in the putative binding site. Complementation with the mutated siaP constructs resulted either in full, partial, or no complementation, depending on the role of specific residues. Mass spectrometry analysis of the O-deacylated LOS of the R127K point mutation confirmed the observation of reduced incorporation of Neu5Ac into the LOS. The decreased ability of H. influenzae to import sialic acid had negative effects on resistance to complement-mediated killing and viability of biofilms in vitro, confirming the importance of sialic acid transport to the bacterium.

  4. Thermodynamics of the interactions of m-AMSA and o-AMSA with nucleic acids: influence of ionic strength and DNA base composition.

    PubMed Central

    Wadkins, R M; Graves, D E

    1989-01-01

    The equilibrium binding of the antitumor agent m-AMSA and its biologically inactive analog o-AMSA to native and synthetic DNAs are compared over a wide range of ionic strengths and temperatures. Although o-AMSA binds DNA with a higher affinity than m-AMSA it is not effective as an antitumor agent. Both m-AMSA and o-AMSA bind DNA in an intercalative manner. Indepth investigations into the thermodynamic parameters of these interactions reveal the interaction of m-AMSA with DNA to be an enthalpy driven process. In contrast, the structurally similar but biologically inactive o-AMSA binds DNA through an entropy driven process. The differences in thermodynamic mechanisms of binding between the two isomers reveal that the electronic and/or steric factors resulting from the position of the methoxy substituent group on the anilino ring directs the DNA binding properties of these compounds and ultimately the biological effectiveness as an antitumor agent. PMID:2602146

  5. A tightly bound quinone functions in the ubiquinone reaction sites of quinoprotein alcohol dehydrogenase of an acetic acid bacterium, Gluconobacter suboxydans.

    PubMed

    Matsushita, Kazunobu; Kobayashi, Yoshiki; Mizuguchi, Mitsuhiro; Toyama, Hirohide; Adachi, Osao; Sakamoto, Kimitoshi; Miyoshi, Hideto

    2008-10-01

    Quinoprotein alcohol dehydrogenase (ADH) of acetic acid bacteria is a membrane-bound enzyme that functions as the primary dehydrogenase in the ethanol oxidase respiratory chain. It consists of three subunits and has a pyrroloquinoline quinone (PQQ) in the active site and four heme c moieties as electron transfer mediators. Of these, three heme c sites and a further site have been found to be involved in ubiquinone (Q) reduction and ubiquinol (QH2) oxidation respectively (Matsushita et al., Biochim. Biophys. Acta, 1409, 154-164 (1999)). In this study, it was found that ADH solubilized and purified with dodecyl maltoside, but not with Triton X-100, had a tightly bound Q, and thus two different ADHs, one having the tightly bound Q (Q-bound ADH) and Q-free ADH, could be obtained. The Q-binding sites of both the ADHs were characterized using specific inhibitors, a substituted phenol PC16 (a Q analog inhibitor) and antimycin A. Based on the inhibition kinetics of Q2 reductase and ubiquinol-2 (Q2H2) oxidase activities, it was suggested that there are one and two PC16-binding sites in Q-bound ADH and Q-free ADH respectively. On the other hand, with antimycin A, only one binding site was found for Q2 reductase and Q2H2 oxidase activities, irrespective of the presence of bound Q. These results suggest that ADH has a high-affinity Q binding site (QH) besides low-affinity Q reduction and QH2 oxidation sites, and that the bound Q in the QH site is involved in the electron transfer between heme c moieties and bulk Q or QH2 in the low-affinity sites.

  6. N-Glycosylation Improves the Pepsin Resistance of Histidine Acid Phosphatase Phytases by Enhancing Their Stability at Acidic pHs and Reducing Pepsin's Accessibility to Its Cleavage Sites.

    PubMed

    Niu, Canfang; Luo, Huiying; Shi, Pengjun; Huang, Huoqing; Wang, Yaru; Yang, Peilong; Yao, Bin

    2015-12-04

    N-Glycosylation can modulate enzyme structure and function. In this study, we identified two pepsin-resistant histidine acid phosphatase (HAP) phytases from Yersinia kristensenii (YkAPPA) and Yersinia rohdei (YrAPPA), each having an N-glycosylation motif, and one pepsin-sensitive HAP phytase from Yersinia enterocolitica (YeAPPA) that lacked an N-glycosylation site. Site-directed mutagenesis was employed to construct mutants by altering the N-glycosylation status of each enzyme, and the mutant and wild-type enzymes were expressed in Pichia pastoris for biochemical characterization. Compared with those of the N-glycosylation site deletion mutants and N-deglycosylated enzymes, all N-glycosylated counterparts exhibited enhanced pepsin resistance. Introduction of the N-glycosylation site into YeAPPA as YkAPPA and YrAPPA conferred pepsin resistance, shifted the pH optimum (0.5 and 1.5 pH units downward, respectively) and improved stability at acidic pH (83.2 and 98.8% residual activities at pH 2.0 for 1 h). Replacing the pepsin cleavage sites L197 and L396 in the immediate vicinity of the N-glycosylation motifs of YkAPPA and YrAPPA with V promoted their resistance to pepsin digestion when produced in Escherichia coli but had no effect on the pepsin resistance of N-glycosylated enzymes produced in P. pastoris. Thus, N-glycosylation may improve pepsin resistance by enhancing the stability at acidic pH and reducing pepsin's accessibility to peptic cleavage sites. This study provides a strategy, namely, the manipulation of N-glycosylation, for improvement of phytase properties for use in animal feed.

  7. N-Glycosylation Improves the Pepsin Resistance of Histidine Acid Phosphatase Phytases by Enhancing Their Stability at Acidic pHs and Reducing Pepsin's Accessibility to Its Cleavage Sites

    PubMed Central

    Niu, Canfang; Luo, Huiying; Shi, Pengjun; Huang, Huoqing; Wang, Yaru; Yang, Peilong

    2015-01-01

    N-Glycosylation can modulate enzyme structure and function. In this study, we identified two pepsin-resistant histidine acid phosphatase (HAP) phytases from Yersinia kristensenii (YkAPPA) and Yersinia rohdei (YrAPPA), each having an N-glycosylation motif, and one pepsin-sensitive HAP phytase from Yersinia enterocolitica (YeAPPA) that lacked an N-glycosylation site. Site-directed mutagenesis was employed to construct mutants by altering the N-glycosylation status of each enzyme, and the mutant and wild-type enzymes were expressed in Pichia pastoris for biochemical characterization. Compared with those of the N-glycosylation site deletion mutants and N-deglycosylated enzymes, all N-glycosylated counterparts exhibited enhanced pepsin resistance. Introduction of the N-glycosylation site into YeAPPA as YkAPPA and YrAPPA conferred pepsin resistance, shifted the pH optimum (0.5 and 1.5 pH units downward, respectively) and improved stability at acidic pH (83.2 and 98.8% residual activities at pH 2.0 for 1 h). Replacing the pepsin cleavage sites L197 and L396 in the immediate vicinity of the N-glycosylation motifs of YkAPPA and YrAPPA with V promoted their resistance to pepsin digestion when produced in Escherichia coli but had no effect on the pepsin resistance of N-glycosylated enzymes produced in P. pastoris. Thus, N-glycosylation may improve pepsin resistance by enhancing the stability at acidic pH and reducing pepsin's accessibility to peptic cleavage sites. This study provides a strategy, namely, the manipulation of N-glycosylation, for improvement of phytase properties for use in animal feed. PMID:26637601

  8. Mass spectrometric identification of citrullination sites and immunohistochemical detection of citrullinated glial fibrillary acidic protein in Alzheimer's disease brains.

    PubMed

    Ishigami, Akihito; Masutomi, Hirofumi; Handa, Setsuko; Nakamura, Megumi; Nakaya, Shuuichi; Uchida, Yoshiaki; Saito, Yuko; Murayama, Shigeo; Jang, Byungki; Jeon, Yong-Chul; Choi, Eun-Kyoung; Kim, Yong-Sun; Kasahara, Yasushi; Maruyama, Naoki; Toda, Tosifusa

    2015-11-01

    Peptidylarginine deiminases (PADs) are posttranslational modification enzymes that convert protein arginine to citrulline residues in a calcium ion-dependent manner. Previously, we reported the abnormal accumulation of citrullinated proteins and the increase in the amount of PAD2 in hippocampi from Alzheimer's disease (AD) patients. Moreover, glial fibrillary acidic protein (GFAP), an astrocyte-specific marker protein, and vimentin were identified as citrullinated proteins by using two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry. To clarify the substrate specificity of PADs against GFAP, we prepared recombinant human (rh)PAD1, rhPAD2, rhPAD3, rhPAD4, and rhGFAP. After incubation of rhGFAP with rhPAD1, rhPAD2, rhPAD3, and rhPAD4, citrullinated (cit-)rhGFAP was detected by Western blotting. The citrullination of rhGFAP by rhPAD2 was unique, specific, and time dependent; additionally, rhPAD1 slightly citrullinated rhGFAP. We then generated eight anti-cit-rhGFAP monoclonal antibodies, CTGF-125, -128, -129, -1212, -1213, -1221, -122R, and -1224R, which reacted specifically with cit-rhGFAP. Two of those eight monoclonal antibodies, CTGF-122R and -1224R, reacted with both cit-rhGFAP and rhGFAP in Western blots. By using the CTGF-1221 antibody and a tandem mass spectrometer, we identified the two independent citrullination sites (R270Cit and R416Cit) of cit-rhGFAP. Immunohistochemical analysis with CTGF-1221 antibody revealed cit-GFAP staining in the hippocampus of AD brain, and the cit-GFAP-positive cells appeared to be astrocyte-like cells. These collective results strongly suggest that PAD2 is responsible for the citrullination of GFAP in the progression of AD and that the monoclonal antibody CTGF-1221, reacting with cit-GFAP at R270Cit and R416Cit, is useful for immunohistochemical investigation of AD brains.

  9. Chemical characterization of the surface sites of coal

    SciTech Connect

    Fowkes, F.M.; Kardos, K.; Riddle, F.L. Jr.; Cole, D.A.

    1990-08-01

    We propose to do experimental studies in four related areas concerning the acid-base properties of coal surfaces: (1) develop high precision flow microcalorimetric methods for determining the concentrations and strengths of the acidic and basic surface sites of coal powders: (2) develop photo-acoustic FTIR and solid-state NMR spectral shift techniques for determination of the concentrations and strengths of acidic and basic surface sites of coal powders; (3) determine the concentrations and strengths of the acidic and basic surface sites of some of the well-characterized coal samples from Argonne National Labs., comparing the coal samples before and after demineralization treatments with HCl and HF; (4) study the effects of surface acidity and basicity on the coal/water interface, with emphasis on the role of interfacial acid-base interactions in the adsorption of ions, surfactants and coal/water slurry stabilizers. From measured heats of interaction, a reasonable estimate can be made of the most prevalent functional groups in coal. This quarter, heats of adsorption of phenols and pyridines were investigated. 2 tabs. (CBS)

  10. Bovine heart fructose-6-phosphate 2-kinase/fructose-2,6-bisphosphatase: complete amino acid sequence and localization of phosphorylation sites.

    PubMed Central

    Sakata, J; Uyeda, K

    1990-01-01

    We have shown previously that bovine heart fructose-6-phosphate 2-kinase/fructose-2,6-bisphosphatase (EC 2.7.1.105/3.1.3.46) is phosphorylated by cAMP-dependent protein kinase and protein kinase C; phosphorylation results in activation of kinase. This activation of heart enzyme is in contrast to results with the liver isozyme, in which phosphorylation by cAMP-dependent protein kinase inhibits the kinase activity. As an initial step toward understanding this difference between the isozymes we have determined the DNA sequence of the heart enzyme and analyzed the amino acid sequence with special emphasis on the location of the phosphorylation site. We isolated and sequenced two overlapping cDNA fragments, which together could encode the complete amino acid sequence of bovine heart fructose-6-phosphate 2-kinase/fructose-2,6-bisphosphatase, a protein of 530 amino acids, with a calculated molecular weight of 60,679. Since the deduced protein contained amino acid sequences identical to the sequences of four known tryptic peptides from this enzyme we concluded that the deduced protein sequence did represent bovine heart enzyme. In addition, a cDNA fragment hybridized to a 4-kilobase mRNA from bovine heart. The phosphorylation sites of the heart enzyme were located near the C terminus, whereas the phosphorylation site of the liver isozyme is known to be located near the N terminus. These opposite locations of the phosphorylation sites may explain the contrasting effect of the covalent modification on the enzymes' activities. Images PMID:2164212

  11. Microbial Community Structure and Physiological Status of Different Types of Biofilms in an Acid Mine Drainage Site Determined by Phospholipid Analysis

    NASA Astrophysics Data System (ADS)

    Fang, J.

    2009-12-01

    A unique aspect of the acid mine drainage (AMD) system at the Green Valley coal mine site (GVS) in western Indiana is the abundance of biofims and biolaminates - stromatolites. Three major types of biofilms have been observed from the AMD site: bright green biofilm dominated by the acidophilic, oxygenic photosynthetic protozoan Euglena mutabilis, olive green biofilm of photosynthetic diatom belonging to the genus Nitzschia, and an olive-green to brownish-green filamentous algae-dominated community. These biofilms are either attached to hard substrata of the effluent channel, or floating at the surface of the effluent with abundant oxygen bubbles, with or without encrusted Fe precipitates. We analyzed lipids (hydrocarbons, wax esters, phospholipids, glycolipids, and neutral lipids) to determine the microbial biomass, community structure and physiological status of biofims collected from the GVS site. Distinctive lipid compositions were observed. The attached, red-crusted biofilms were characterized by abundant wax esters, monounsaturated fatty acids, whereas the floating biofilms by phytadienes, phytanol, polyunsaturated n-alkenes, polyunsaturated fatty acids. The accumulation of abundant wax esters probably reflects the readily available carbon and limitation of nutrients to the biofilm. Alternatively, the wax esters may be the biochemical relics of the anaerobic past of the Earth and the detection of these compounds has important implications for the evolution of eukaryotes and the paleo-environmental conditions on early Earth. This type of biochemical machine may have allowed early eukaryotes to survive recurrent anoxic conditions on early Earth.

  12. The reaction mechanism for the SCR process on monomer V(5+) sites and the effect of modified Brønsted acidity.

    PubMed

    Arnarson, Logi; Falsig, Hanne; Rasmussen, Søren B; Lauritsen, Jeppe V; Moses, Poul Georg

    2016-06-22

    The energetics, structures and activity of a monomeric VO3H/TiO2(001) catalyst are investigated for the selective catalytic reduction (SCR) reaction by the use of density functional theory (DFT). Furthermore we study the influences of a dopant substitute in the TiO2 support and its effects on the known properties of the SCR system such as Brønsted acidity and reducibility of vanadium. We find for the reduction part of the SCR mechanism that it involves two Ti-O-V oxygen sites. One is a hydroxyl possessing Brønsted acidity which contributes to the formation of NH4(+), while the other accepts a proton which charge stabilizes the reduced active site. In the reduction the proton is donated to the latter due to a reaction between NH3 and NO that forms a H2NNO molecule which decomposes into N2(g) and H2O(g). A dopant substitution of 10 different dopants: Si, Ge, Se, Zr, Sn, Te, Hf, V, Mo and W at each of the sites, which participate in the reaction, modifies the energetics and therefore the SCR activity. We find that Brønsted acidity is a descriptor for the SCR activity at low temperatures. Based on this descriptor we find that Zr, Hf and Sn have a positive effect as they decrease the activation energy for the SCR reaction. PMID:27297567

  13. A quantum chemical study for exploring the inhibitory effect of nitrogen containing species on the adsorption of polynuclear aromatic hydrocarbons over a Bronsted acid site

    NASA Astrophysics Data System (ADS)

    Celis-Cornejo, C. M.; Garnica Mantilla, M. M.; Baldovino-Medrano, V. G.; Ramírez-Caballero, G. E.

    2016-08-01

    The analysis of the inhibitory effect of nitrogenated compounds on the hydroprocessing and hydropurification of oil derived fuels is important to produce cleaner fuels. In this work, density functional theory calculations were performed to investigate the effect of the nitrogen containing molecules on the adsorption of Polynuclear Aromatic Hydrocarbons (PAHs). Mordenite was chosen as a zeolitic structure for simulating a Bronsted acid site. The character of the acid site was confirmed by both a vibrational frequency calculation and a Bader charge analysis. From the adsorption calculations, it was found that the adsorption energy of PAHs increases with the number of aromatic rings in the structure. Also, the nitrogen containing species possibly inhibit more extensively two and three rings PAHs because of their lower adsorption energies. Finally, it was observed that the nitrogen species tend to drag the proton from the mordenite acid site. This explains the inhibitory effect in the adsorption of PAHs and contributes to understanding the dynamics of hydrocarbon hydroprocessing in refineries.

  14. The reaction mechanism for the SCR process on monomer V(5+) sites and the effect of modified Brønsted acidity.

    PubMed

    Arnarson, Logi; Falsig, Hanne; Rasmussen, Søren B; Lauritsen, Jeppe V; Moses, Poul Georg

    2016-06-22

    The energetics, structures and activity of a monomeric VO3H/TiO2(001) catalyst are investigated for the selective catalytic reduction (SCR) reaction by the use of density functional theory (DFT). Furthermore we study the influences of a dopant substitute in the TiO2 support and its effects on the known properties of the SCR system such as Brønsted acidity and reducibility of vanadium. We find for the reduction part of the SCR mechanism that it involves two Ti-O-V oxygen sites. One is a hydroxyl possessing Brønsted acidity which contributes to the formation of NH4(+), while the other accepts a proton which charge stabilizes the reduced active site. In the reduction the proton is donated to the latter due to a reaction between NH3 and NO that forms a H2NNO molecule which decomposes into N2(g) and H2O(g). A dopant substitution of 10 different dopants: Si, Ge, Se, Zr, Sn, Te, Hf, V, Mo and W at each of the sites, which participate in the reaction, modifies the energetics and therefore the SCR activity. We find that Brønsted acidity is a descriptor for the SCR activity at low temperatures. Based on this descriptor we find that Zr, Hf and Sn have a positive effect as they decrease the activation energy for the SCR reaction.

  15. Site-directed mutagenesis of highly conserved amino acids in the first cytoplasmic loop of Drosophila Rh1 opsin blocks rhodopsin synthesis in the nascent state.

    PubMed Central

    Bentrop, J; Schwab, K; Pak, W L; Paulsen, R

    1997-01-01

    The cytoplasmic surface of Drosophila melanogaster Rh1 rhodopsin (ninaE) harbours amino acids which are highly conserved among G-protein-coupled receptors. Site-directed mutations which cause Leu81Gln or Asn86Ile amino acid substitutions in the first cytoplasmic loop of the Rh1 opsin protein, are shown to block rhodopsin synthesis in the nascent, glycosylated state from which the mutant opsin is degraded rapidly. In mutants Leu81Gln and Asn86Ile, only 20-30% and <2% respectively, of functional rhodopsins are synthesized and transported to the photoreceptive membrane. Thus, conserved amino acids in opsin's cytoplasmic surface are a critical factor in the interaction of opsin with proteins of the rhodopsin processing machinery. Photoreceptor cells expressing mutant rhodopsins undergo age-dependent degeneration in a recessive manner. PMID:9130705

  16. pH- and ionic-strength-induced structural changes in poly(acrylic acid)-lipid-based self-assembled materials.

    SciTech Connect

    Crisci, A.; Hay, D. N. T.; Seifert, S.; Firestone, M. A.

    2009-01-01

    The effect of a polyanion introduced as a lipid conjugate (poly(acrylic acid)- dimyristoyl-sn-glycero-3-phosphoethanolamine, PAA-DMPE) on the structure of a self-assembled, biomembrane mimetic has been evaluated using synchrotron small-angle X-ray scattering (SAXS). At high grafting density (8-11 mol.%), the PAA chains were found to produce significant changes in structure in response to changes in pH and electrolyte composition. At low pH and in the absence of salt (NaCl), the neutral PAA chains adopt a coil conformational state that leads to the formation of a swollen lamellar structure. Upon the addition of salt at low to intermediate pH values, two lamellar phases, a collapsed and an expanded structure, coexist. Finally, when the polymer is fully ionized (at high pH), the extended conformation of the polymer generates a cubic phase. The results of this study contribute to an understanding of how polyelectrolytes may ultimately be harnessed for the preparation of self-assembling materials responsive to external stimuli.

  17. Identification of Active and Spectator Sn Sites in Sn-β Following Solid-State Stannation, and Consequences for Lewis Acid Catalysis

    PubMed Central

    Hammond, Ceri; Padovan, Daniele; Al-Nayili, Abbas; Wells, Peter P; Gibson, Emma K; Dimitratos, Nikolaos

    2015-01-01

    Lewis acidic zeolites are rapidly emerging liquid-phase Lewis acid catalysts. Nevertheless, their inefficient synthesis procedure currently prohibits greater utilization and exploitation of these promising materials. Herein, we demonstrate that SnIV-containing zeolite beta can readily be prepared both selectively and extremely rapidly by solid-state incorporation (SSI) method. Through a combination of spectroscopic (XRD, UV/Vis, X-ray absorption, magic-angle spinning NMR, and diffuse reflectance infrared Fourier transform spectroscopy) studies, we unambiguously demonstrate that site-isolated, isomorphously substituted SnIV sites dominate the Sn population up to a loading of 5 wt % Sn. These sites are identical to those found in conventionally prepared Sn-beta, and result in our SSI material exhibiting identical levels of intrinsic activity (that is, turnover frequency) despite the threefold increase in Sn loading, and the extremely rapid and benign nature of our preparation methodology. We also identify the presence of spectator sites, in the form of SnIV oligomers, at higher levels of Sn loading. The consequences of this mixed population with regards to catalysis (Meerwein–Pondorf–Verley reaction and glucose isomerization) are also identified. PMID:26583051

  18. Multiple Propofol-binding Sites in a γ-Aminobutyric Acid Type A Receptor (GABAAR) Identified Using a Photoreactive Propofol Analog*♦

    PubMed Central

    Jayakar, Selwyn S.; Zhou, Xiaojuan; Chiara, David C.; Dostalova, Zuzana; Savechenkov, Pavel Y.; Bruzik, Karol S.; Dailey, William P.; Miller, Keith W.; Eckenhoff, Roderic G.; Cohen, Jonathan B.

    2014-01-01

    Propofol acts as a positive allosteric modulator of γ-aminobutyric acid type A receptors (GABAARs), an interaction necessary for its anesthetic potency in vivo as a general anesthetic. Identifying the location of propofol-binding sites is necessary to understand its mechanism of GABAAR modulation. [3H]2-(3-Methyl-3H-diaziren-3-yl)ethyl 1-(phenylethyl)-1H-imidazole-5-carboxylate (azietomidate) and R-[3H]5-allyl-1-methyl-5-(m-trifluoromethyl-diazirynylphenyl)barbituric acid (mTFD-MPAB), photoreactive analogs of 2-ethyl 1-(phenylethyl)-1H-imidazole-5-carboxylate (etomidate) and mephobarbital, respectively, have identified two homologous but pharmacologically distinct classes of intersubunit-binding sites for general anesthetics in the GABAAR transmembrane domain. Here, we use a photoreactive analog of propofol (2-isopropyl-5-[3-(trifluoromethyl)-3H-diazirin-3-yl]phenol ([3H]AziPm)) to identify propofol-binding sites in heterologously expressed human α1β3 GABAARs. Propofol, AziPm, etomidate, and R-mTFD-MPAB each inhibited [3H]AziPm photoincorporation into GABAAR subunits maximally by ∼50%. When the amino acids photolabeled by [3H]AziPm were identified by protein microsequencing, we found propofol-inhibitable photolabeling of amino acids in the β3-α1 subunit interface (β3Met-286 in β3M3 and α1Met-236 in α1M1), previously photolabeled by [3H]azietomidate, and α1Ile-239, located one helical turn below α1Met-236. There was also propofol-inhibitable [3H]AziPm photolabeling of β3Met-227 in βM1, the amino acid in the α1-β3 subunit interface photolabeled by R-[3H]mTFD-MPAB. The propofol-inhibitable [3H]AziPm photolabeling in the GABAAR β3 subunit in conjunction with the concentration dependence of inhibition of that photolabeling by etomidate or R-mTFD-MPAB also establish that each anesthetic binds to the homologous site at the β3-β3 subunit interface. These results establish that AziPm as well as propofol bind to the homologous intersubunit sites in the

  19. Influence of surface treatments on the shear bond strength of orthodontic brackets to porcelain

    NASA Astrophysics Data System (ADS)

    Wang, Cong; Zeng, Jishan; Wang, Shaoan; Yang, Zheng; Huang, Qian; Chen, Pixiu; Zhou, Shujuan; Liu, Xiaoqing

    2008-11-01

    The purpose of this study was to investigate the effect of various surface treatments after different storage time and thermocycling on the shear bond strength of orthodontic brackets to the feldspathic porcelain surfaces. 128 disc-shaped porcelain specimens were randomly assigned to the following surface treatments: 9.6% HFA, 9.6% HFA combined with silane, 50 μ aluminum trioxide sandblasting followed by silane and application of silane after 37% phosphoric acid. Metal or ceramic brackets were bonded onto each treated porcelain facet with light cured resin. The samples were stored in 37 °C water 1 day or 7 days, thermocycled 500 times from 5 to 55 °C. The shear bond strengths were measured (1 mm/min), and statistically analyzed. The bond failure sites were classified according to ARI system. The surface of the glazed, sandblasted, hydrofluoric and phosphoric acid etched porcelain were examined with SEM. All groups achieved reasonable bond strengths to withstand the application of orthodontic forces. Water storage for 7 days caused lower shear bond strength than that of 1 day. But there is no statistically significant difference between the two groups. The mean shear bond strength provided by ceramic bracket with mechanical retention had no statistical difference with that of metal bracket. Therefore, the optimal treatment for orthodontic brackets bonding to feldspathic porcelain was to apply phosphoric acid combined with silane.

  20. Changes in the Cytoplasmic Composition of Amino Acids and Proteins Observed in Staphylococcus aureus during Growth under Variable Growth Conditions Representative of the Human Wound Site

    PubMed Central

    Alreshidi, Mousa M.; Dunstan, R. Hugh; Gottfries, Johan; Macdonald, Margaret M.; Crompton, Marcus J.; Ang, Ching-Seng; Williamson, Nicholas A.; Roberts, Tim K.

    2016-01-01

    Staphylococcus aureus is an opportunistic pathogen responsible for a high proportion of nosocomial infections. This study was conducted to assess the bacterial responses in the cytoplasmic composition of amino acids and ribosomal proteins under various environmental conditions designed to mimic those on the human skin or within a wound site: pH6-8, temperature 35–37°C, and additional 0–5% NaCl. It was found that each set of environmental conditions elicited substantial adjustments in cytoplasmic levels of glutamic acid, aspartic acid, proline, alanine and glycine (P< 0.05). These alterations generated characteristic amino acid profiles assessed by principle component analysis (PCA). Substantial alterations in cytoplasmic amino acid and protein composition occurred during growth under conditions of higher salinity stress implemented via additional levels of NaCl in the growth medium. The cells responded to additional NaCl at pH 6 by reducing levels of ribosomal proteins, whereas at pH 8 there was an upregulation of ribosomal proteins compared with the reference control. The levels of two ribosomal proteins, L32 and S19, remained constant across all experimental conditions. The data supported the hypothesis that the bacterium was continually responding to the dynamic environment by modifying the proteome and optimising metabolic homeostasis. PMID:27442022

  1. Active-Site Engineering of ω-Transaminase for Production of Unnatural Amino Acids Carrying a Side Chain Bulkier than an Ethyl Substituent

    PubMed Central

    Han, Sang-Woo; Park, Eul-Soo; Dong, Joo-Young

    2015-01-01

    ω-Transaminase (ω-TA) is a promising enzyme for use in the production of unnatural amino acids from keto acids using cheap amino donors such as isopropylamine. The small substrate-binding pocket of most ω-TAs permits entry of substituents no larger than an ethyl group, which presents a significant challenge to the preparation of structurally diverse unnatural amino acids. Here we report on the engineering of an (S)-selective ω-TA from Ochrobactrum anthropi (OATA) to reduce the steric constraint and thereby allow the small pocket to readily accept bulky substituents. On the basis of a docking model in which l-alanine was used as a ligand, nine active-site residues were selected for alanine scanning mutagenesis. Among the resulting variants, an L57A variant showed dramatic activity improvements in activity for α-keto acids and α-amino acids carrying substituents whose bulk is up to that of an n-butyl substituent (e.g., 48- and 56-fold increases in activity for 2-oxopentanoic acid and l-norvaline, respectively). An L57G mutation also relieved the steric constraint but did so much less than the L57A mutation did. In contrast, an L57V substitution failed to induce the improvements in activity for bulky substrates. Molecular modeling suggested that the alanine substitution of L57, located in a large pocket, induces an altered binding orientation of an α-carboxyl group and thereby provides more room to the small pocket. The synthetic utility of the L57A variant was demonstrated by carrying out the production of optically pure l- and d-norvaline (i.e., enantiomeric excess [ee] > 99%) by asymmetric amination of 2-oxopantanoic acid and kinetic resolution of racemic norvaline, respectively. PMID:26231640

  2. Site-specific solvent exposure analysis of a membrane protein using unnatural amino acids and {sup 19}F nuclear magnetic resonance

    SciTech Connect

    Shi, Pan; Li, Dong; Chen, Hongwei; Xiong, Ying; Tian, Changlin

    2011-10-22

    Highlights: {yields} Solvent isotope shift analysis of {sup 19}F-tfmF in different H{sub 2}O/D{sub 2}O molar ratio. {yields} Correlation between solvent isotope shift of {sup 19}F-spins and solvent exposure analysis. {yields} Solvent exposure analysis of membrane proteins. -- Abstract: Membrane proteins play an essential role in cellular metabolism, transportation and signal transduction across cell membranes. The scarcity of membrane protein structures has thus far prevented a full understanding of their molecular mechanisms. Preliminary topology studies and residue solvent exposure analysis have the potential to provide valuable information on membrane proteins of unknown structure. Here, a {sup 19}F-containing unnatural amino acid (trimethylfluoro-phenylalanine, tfmF) was applied to accomplish site-specific {sup 19}F spin incorporation at different sites in diacylglycerol kinase (DAGK, an Escherichia coli membrane protein) for site-specific solvent exposure analysis. Due to isotope effect on {sup 19}F spins, a standard curve for {sup 19}F-tfmF chemical shifts was drawn for varying solvent H{sub 2}O/D{sub 2}O ratios. Further site-specific {sup 19}F solvent isotope shift analysis was conducted for DAGK to distinguish residues in water-soluble loops, interfacial areas or hydrophobic membrane regions. This site-specific solvent exposure analysis method could be applied for further topological analysis of other membrane proteins.

  3. Acidic episodes and surface-water chemistry: a comparison of northeast and southeast study sites. Project report

    SciTech Connect

    Ford, D.E.; Malcom, J.T.; Murdoch, P.S.; Olem, H.; Witt, E.C.

    1987-05-01

    Much of the emphasis in the National Acid Precipitation Assessment Program (NAPAP) has been on historical or long-term trends in surface-water acidification. Short-term acidic episodes, however, also might have significant adverse effects on aquatic ecosystems. The U.S. EPA is presently designing an Episodic Response Project to investigate the regional extent, frequency, duration and magnitude of acidic episodes. The studies discussed in the report, however, were conducted under NAPAP Task Group E2-Aquatic Effects. A total of four episodic studies were conducted in the Catskill Mountains of New York (Murdoch, USGS), Laurel Hills, PA (Witt and Barker, USGS), Southern Blue Ridge Province, NC, TN (Olem, TVA), and Ouachita Mountains, AR (Nix et al., Ouachita Baptist University).

  4. In vivo biotinylation and incorporation of a photo-inducible unnatural amino acid to an antibody-binding domain improve site-specific labeling of antibodies.

    PubMed

    Kanje, Sara; Hober, Sophia

    2015-04-01

    Antibodies are important molecules in many research fields, where they play a key role in various assays. Antibody labeling is therefore of great importance. Currently, most labeling techniques take advantage of certain amino acid side chains that commonly appear throughout proteins. This makes it hard to control the position and exact degree of labeling of each antibody. Hence, labeling of the antibody may affect the antibody-binding site. This paper presents a novel protein domain based on the IgG-binding domain C2 of streptococcal protein G, containing the unnatural amino acid BPA, that can cross-link other molecules. This novel domain can, with improved efficiency compared to previously reported similar domains, site-specifically cross-link to IgG at the Fc region. An efficient method for simultaneous in vivo incorporation of BPA and specific biotinylation in a flask cultivation of Escherichia coli is described. In comparison to a traditionally labeled antibody sample, the C2-labeled counterpart proved to have a higher proportion of functional antibodies when immobilized on a solid surface and the same limit of detection in an ELISA. This method of labeling is, due to its efficiency and simplicity, of high interest for all antibody-based assays where it is important that labeling does not interfere with the antibody-binding site.

  5. Probing the S-adenosylmethionine-binding site of rat guanidinoacetate methyltransferase. Effect of site-directed mutagenesis of residues that are conserved across mammalian non-nucleic acid methyltransferases.

    PubMed Central

    Hamahata, A; Takata, Y; Gomi, T; Fujioka, M

    1996-01-01

    Most mammalian non-nucleic acid methyltransferases share three sequence motifs. To gain insight into the S-adenosyl-methionine (AdoMet)-binding site of guanidinoacetate methyltransferase, we mutated several conserved residues that are found in or near motifs I and II. Conversion of either of two glycine residues of motif I (Gly67 and Gly69) to an alanine resulted in an inactive enzyme. These enzymes, although having UV absorption, fluorescence and far-UV CD spectra virtually identical with those of the wild-type enzyme, seem to be conformationally different from the wild-type enzyme as judged by near-UV CD spectra and the extent of urea denaturation, and are apparently not capable of binding AdoMet. Mutation of Tyr136 of motif II to a valine resulted in a decrease in Kcat/Km values for substrates. Changing this residue to a phenylalanine caused only a minor change in Kcat/Km for AdoMet. This suggests that the aromatic side chain stabilizes the binding of AdoMet. Mutagenic changes of Glu89, which is the residue corresponding to the conserved acidic residue on the C-terminal side of motif I, indicated its contribution to AdoMet binding. These results are consistent with the idea that both motifs I and II are crucial in forming the AdoMet binding site of guanidinoacetate methyltransferase. PMID:8694756

  6. Potential use of fatty acid profiles of the adductor muscle of cockles (Cerastoderma edule) for traceability of collection site.

    PubMed

    Ricardo, Fernando; Pimentel, Tânia; Moreira, Ana S P; Rey, Felisa; Coimbra, Manuel A; Rosário Domingues, M; Domingues, Pedro; Costa Leal, Miguel; Calado, Ricardo

    2015-06-18

    Geographic traceability of seafood is key for controlling its quality and safeguarding consumers' interest. The present study assessed if the fatty acid (FA) profile of the adductor muscle (AM) of fresh cockles (Cerastoderma edule) can be used to discriminate the origin of specimens collected in different bivalve capture/production areas legally defined within a coastal lagoon. Results suggest that this biochemical approach holds the potential to trace sampling locations with a spatial resolution <10 Km, even for areas with identical classification for bivalve production. Cockles further away from the inlet, i.e. in areas exposed to a higher saline variation, exhibited lower levels of saturated fatty acids, which are key for stabilizing the bilayer structure of cell membranes, and a higher percentage of polyunsaturated fatty acids, which enhance bilayer fluidity. Results suggest that the structural nature of the lipids present in the AM provides a stable fatty acid signature and holds potential for tracing the origin of bivalves to their capture/production areas.

  7. Particle-facilitated lead and arsenic transport in abandoned mine sites soil influenced by simulated acid rain.

    PubMed

    Shaoping, Hu; Xincai, Chen; Jiyan, Shi; Yingxu, Chen; Qi, Lin

    2008-05-01

    The role of acid rain in affecting Pb and As transport from mine tailings was investigated by pumping simulated acid rain at a infiltration rate of 10.2 cm/h through soil columns. Simulated acid rain with pH of 3.0, 4.5 and 5.6 were used as leaching solutions. Results showed that 86.9-95.9% of Pb and 90-91.8% of As eluted from the columns were adsorbed by particles in the leachates. Scanning electron microscopy (SEM) analysis showed that particles released from the columns were mainly composed of flocculated aggregates and plate or rod shaped discrete grains. Transmission electron microscopy (TEM) coupled with energy dispersive X-ray analysis (EDX) showed that these particles were predominantly silicate minerals. Results from our experiments demonstrated that when rapid infiltration conditions or a rainstorm exist, particle-facilitated transport of contaminants is likely to the dominant metal transport pathway influenced by acid rain.

  8. The RRM Domain of Human Fused in Sarcoma Protein Reveals a Non-Canonical Nucleic Acid Binding Site

    PubMed Central

    Liu, Xuehui; Niu, Chunyan; Ren, Jintao; Zhang, Jiayu; Xie, Xiaodong; Zhu, Haining; Feng, Wei; Gong, Weimin

    2012-01-01

    Fused in sarcoma (FUS) is involved in many processes of RNA metabolism. FUS and another RNA binding protein, TDP-43, are implicated in amyotrophic lateral sclerosis (ALS). It is significant to characterize the RNA recognition motif (RRM) of FUS as its nucleic acid binding properties are unclear. More importantly, abolishing the RNA binding ability of the RRM domain of TDP43 was reported to suppress the neurotoxicity of TDP-43 in Drosophila. The sequence of FUS-RRM varies significantly from canonical RRMs, but the solution structure of FUS-RRM determined by NMR showed a similar overall folding as other RRMs. We found that FUS-RRM directly bound to RNA and DNA and the binding affinity was in the micromolar range as measured by surface plasmon resonance and NMR titration. The nucleic acid binding pocket in FUS-RRM is significantly distorted since several critical aromatic residues are missing. An exceptionally positively charged loop in FUS-RRM, which is not found in other RRMs, is directly involved in the RNA/DNA binding. Substituting the lysine residues in the unique KK loop impaired the nucleic acid binding and altered FUS subcellular localization. The results provide insights into the nucleic acid binding properties of FUS-RRM and its potential relevance to ALS. PMID:23200923

  9. Potential use of fatty acid profiles of the adductor muscle of cockles (Cerastoderma edule) for traceability of collection site.

    PubMed

    Ricardo, Fernando; Pimentel, Tânia; Moreira, Ana S P; Rey, Felisa; Coimbra, Manuel A; Rosário Domingues, M; Domingues, Pedro; Costa Leal, Miguel; Calado, Ricardo

    2015-01-01

    Geographic traceability of seafood is key for controlling its quality and safeguarding consumers' interest. The present study assessed if the fatty acid (FA) profile of the adductor muscle (AM) of fresh cockles (Cerastoderma edule) can be used to discriminate the origin of specimens collected in different bivalve capture/production areas legally defined within a coastal lagoon. Results suggest that this biochemical approach holds the potential to trace sampling locations with a spatial resolution <10 Km, even for areas with identical classification for bivalve production. Cockles further away from the inlet, i.e. in areas exposed to a higher saline variation, exhibited lower levels of saturated fatty acids, which are key for stabilizing the bilayer structure of cell membranes, and a higher percentage of polyunsaturated fatty acids, which enhance bilayer fluidity. Results suggest that the structural nature of the lipids present in the AM provides a stable fatty acid signature and holds potential for tracing the origin of bivalves to their capture/production areas. PMID:26084395

  10. Potential use of fatty acid profiles of the adductor muscle of cockles (Cerastoderma edule) for traceability of collection site

    PubMed Central

    Ricardo, Fernando; Pimentel, Tânia; Moreira, Ana S. P.; Rey, Felisa; Coimbra, Manuel A.; Rosário Domingues, M.; Domingues, Pedro; Costa Leal, Miguel; Calado, Ricardo

    2015-01-01

    Geographic traceability of seafood is key for controlling its quality and safeguarding consumers’ interest. The present study assessed if the fatty acid (FA) profile of the adductor muscle (AM) of fresh cockles (Cerastoderma edule) can be used to discriminate the origin of specimens collected in different bivalve capture/production areas legally defined within a coastal lagoon. Results suggest that this biochemical approach holds the potential to trace sampling locations with a spatial resolution <10 Km, even for areas with identical classification for bivalve production. Cockles further away from the inlet, i.e. in areas exposed to a higher saline variation, exhibited lower levels of saturated fatty acids, which are key for stabilizing the bilayer structure of cell membranes, and a higher percentage of polyunsaturated fatty acids, which enhance bilayer fluidity. Results suggest that the structural nature of the lipids present in the AM provides a stable fatty acid signature and holds potential for tracing the origin of bivalves to their capture/production areas. PMID:26084395

  11. Flexibility and Muscular Strength.

    ERIC Educational Resources Information Center

    Liemohn, Wendell

    1988-01-01

    This definition of flexibility and muscular strength also explores their roles in overall physical fitness and focuses on how increased flexibility and muscular strength can help decrease or eliminate lower back pain. (CB)

  12. Non-coding nucleotides and amino acids near the active site regulate peptide deformylase expression and inhibitor susceptibility in Chlamydia trachomatis

    PubMed Central

    Bao, Xiaofeng; Pachikara, Niseema D.; Oey, Christopher B.; Balakrishnan, Amit; Westblade, Lars F.; Tan, Ming; Chase, Theodore; Nickels, Bryce E.

    2011-01-01

    Chlamydia trachomatis, an obligate intracellular bacterium, is a highly prevalent human pathogen. Hydroxamic-acid-based matrix metalloprotease inhibitors can effectively inhibit the pathogen both in vitro and in vivo, and have exhibited therapeutic potential. Here, we provide genome sequencing data indicating that peptide deformylase (PDF) is the sole target of the inhibitors in this organism. We further report molecular mechanisms that control chlamydial PDF (cPDF) expression and inhibition efficiency. In particular, we identify the σ66-dependent promoter that controls cPDF gene expression and demonstrate that point mutations in this promoter lead to resistance by increasing cPDF transcription. Furthermore, we show that substitution of two amino acids near the active site of the enzyme alters enzyme kinetics and protein stability. PMID:21719536

  13. MERCURY-NITRITE-RHODIUM-RUTHENIUM INTERACTIONS IN NOBLE METAL CATALYZED HYDROGEN GENERATION FROM FORMIC ACID DURING NUCLEAR WASTE PROCESSING AT THE SAVANNAH RIVER SITE - 136C

    SciTech Connect

    Koopman, D.; Pickenheim, B.; Lambert, D.; Newell, J; Stone, M.

    2009-09-02

    Chemical pre-treatment of radioactive waste at the Savannah River Site is performed to prepare the waste for vitrification into a stable waste glass form. During pre-treatment, compounds in the waste become catalytically active. Mercury, rhodium, and palladium become active for nitrite destruction by formic acid, while rhodium and ruthenium become active for catalytic conversion of formic acid into hydrogen and carbon dioxide. Nitrite ion is present during the maximum activity of rhodium, but is consumed prior to the activation of ruthenium. Catalytic hydrogen generation during pre-treatment can exceed radiolytic hydrogen generation by several orders of magnitude. Palladium and mercury impact the maximum catalytic hydrogen generation rates of rhodium and ruthenium by altering the kinetics of nitrite ion decomposition. New data are presented that illustrate the interactions of these various species.

  14. Measurement of caffeic and ferulic acid equivalents in plasma after coffee consumption: small intestine and colon are key sites for coffee metabolism.

    PubMed

    Renouf, Mathieu; Guy, Philippe A; Marmet, Cynthia; Fraering, Anne-Lise; Longet, Karin; Moulin, Julie; Enslen, Marc; Barron, Denis; Dionisi, Fabiola; Cavin, Christophe; Williamson, Gary; Steiling, Heike

    2010-06-01

    Previous studies on coffee examined absorption of phenolic acids (PA) in the small intestine, but not the contribution of the colon to absorption. Nine healthy volunteers ingested instant soluble coffee ( approximately 335 mg total chlorogenic acids (CGAs)) in water. Blood samples were taken over 12 h, and at 24 h to assess return to baseline. Many previous studies, which used glucuronidase and sulfatase, measured only PA and did not rigorously assess CGAs. To improve this, plasma samples were analyzed after full hydrolysis by chlorogenate esterase, glucuronidase and sulfatase to release aglycone equivalents of PA followed by liquid-liquid extraction and ESI-LC-ESI-MS/MS detection. Ferulic, caffeic and isoferulic acid equivalents appeared rapidly in plasma, peaking at 1-2 h. Dihydrocaffeic and dihydroferulic acids appeared in plasma 6-8 h after ingestion (T(max=)8-12 h). Substantial variability in maximum plasma concentration and T(max) was also observed between individuals. This study confirms that the small intestine is a significant site for absorption of PA, but shows for the first time that the colon/microflora play the major role in absorption and metabolism of CGAs and PA from coffee. PMID:19937852

  15. Incorporation and electron transfer of anthracene in pores of ZSM-5 zeolites. Effect of Brønsted acid site density.

    PubMed

    Hureau, Matthieu; Moissette, Alain; Marquis, Séverine; Brémard, Claude; Vezin, Hervé

    2009-08-01

    The sorption course of anthracene (ACENE-3) into Brønsted-acidic medium pore MFI zeolites was monitored by in situ EPR and diffuse reflectance UV-visible absorption over one year. Weighed amounts of solid ACENE-3 were merely exposed to H(n)ZSM-5 (H(n)(AlO(2))(n)(SiO(2))(96-n)), with the following Brønsted acid site (BAS) densities, n = 0.0, 0.17, 0.57, 0.95, 2.0, 3.4, 6.6, dehydrated at 623 K under argon. The weighed amounts correspond to 1 ACENE-3 per zeolite unit cell. ACENE-3 is found to be incorporated as intact molecules in purely siliceous MFI (silicalite-1). Monte Carlo simulations indicate that ACENE-3 lies in the intersection of straight and zigzag channels. In contrast, the presence of BASs on the inner surface of channels induces spontaneous ionization of ACENE-3 (ionization potential = 7.44 eV). The charge separation as ACENE-3*(+)@H(n)ZSM-5*(-) is caused by the strong Coulombic field gradient of Si-O(-)(H(+))-Al BAS in the absence of any Lewis acid site. The rate and yield of ionization are found to increase dramatically with BAS density increase. The stabilization of ACENE-3*(+)@H(n)ZSM-5*(-) is explained by the tight fit between the rod-shape ACENE-3 and the channel dimensions and especially by the compartmentalization of ejected electrons as AlO(4)H*(-) centers away from the initial site of ionization. The final charge recombination occurs after more than one year and leads to ACENE-3 occluded in the straight channel in close proximity to BAS without any protonation of ACENE-3 (pK(a) = -13.5).

  16. Hydroxamic acid interactions with solvated cerium hydroxides in the flotation of monazite and bastnäsite-Experiments and DFT study

    NASA Astrophysics Data System (ADS)

    Sarvaramini, A.; Azizi, D.; Larachi, F.

    2016-11-01

    Density functional theory (DFT) simulations and experiments were performed to clarify the interaction mechanisms between hydroxamic acid collectors and cerium hydroxides during the flotation of bastnäsite and monazite minerals. These minerals showed considerable floatability at moderately alkaline pH which was related to the adsorption of hydroxamic acids on their surfaces as confirmed by vibrational spectroscopic and zeta potential measurements. DFT simulations showed that at moderately alkaline pH, the interactions between solvated Ce(OH)2+ and Ce(OH)2+ and heptyl-hydroxamic acid (HHA) anions resulted in the formation of, respectively, [Ce(OH)(HHA)x(H2O)y]2-x (x[y = ] = 1[6],2[3],3[1]) and [Ce(OH)2(HHA)x(H2O)y]1-x (x[y = ] = 1[5],2[1],3[0]) complexes. The collector anions were found to interact directly through formation of two covalent bonds between their two polar-head oxygen atoms and cerium in the hydroxide complexes. However, formation of such new bonds resulted in breakage of a few covalent/electrostatic bonds between cerium and water molecules initially present in the first hydration shell of the rare-earth metal cation. Building up in the electric double layer of the semi-soluble minerals, these complexes, and by extension, those from other rare-earth elements belonging to monazite and bastnäsite, are speculated to play a role in the interactions between rare-earth minerals and hydroxamic acid collectors.

  17. What Is a Strength?

    ERIC Educational Resources Information Center

    Wolin, Sybil

    2003-01-01

    As the strength-based perspective gains recognition, it is important to describe what constitutes strengths and to develop a specific vocabulary to name them. This article draws on resilience research to help identify specific competencies and areas of strengths in youth. (Contains 1 table.)

  18. Strength Training for Girls.

    ERIC Educational Resources Information Center

    Connaughton, Daniel; Connaughton, Angela; Poor, Linda

    2001-01-01

    Strength training can be fun, safe, and appropriate for young girls and women and is an important component of any fitness program when combined with appropriate cardiovascular and flexibility activities. Concerns and misconceptions regarding girls' strength training are discussed, presenting general principles of strength training for children…

  19. Directing Group in Decarboxylative Cross-Coupling: Copper-Catalyzed Site-Selective C-N Bond Formation from Nonactivated Aliphatic Carboxylic Acids.

    PubMed

    Liu, Zhao-Jing; Lu, Xi; Wang, Guan; Li, Lei; Jiang, Wei-Tao; Wang, Yu-Dong; Xiao, Bin; Fu, Yao

    2016-08-01

    Copper-catalyzed directed decarboxylative amination of nonactivated aliphatic carboxylic acids is described. This intramolecular C-N bond formation reaction provides efficient access to the synthesis of pyrrolidine and piperidine derivatives as well as the modification of complex natural products. Moreover, this reaction presents excellent site-selectivity in the C-N bond formation step through the use of directing group. Our work can be considered as a big step toward controllable radical decarboxylative carbon-heteroatom cross-coupling. PMID:27439145

  20. Mixed-Metal-Organic Framework with Effective Lewis Acidic Sites for Sulfur Confinement in High-Performance Lithium-Sulfur Batteries.

    PubMed

    Wang, Ziqi; Wang, Buxue; Yang, Yu; Cui, Yuanjing; Wang, Zhiyu; Chen, Banglin; Qian, Guodong

    2015-09-23

    The mixed-metal-organic framework approach and a representative zirconium-metalloporphyrin framework (MOF-525) have been developed to create novel sulfur hosts and Li-S batteries. The different local environments at the centers of the porphyrin moieties in a series of MMOFs-MOF-525(2H), MOF-525(FeCl), and MOF-525(Cu)-have led to their different behaviors for the confinement of sulfur and thus Li-S batteries. The unique structure of MOF-525(Cu) has enabled each Cu(2+) site to offer two Lewis acidic sites, featuring it as a very powerful MOF host for the inclusion of sulfur and polysulfides. The S@MOF-525(Cu) cathode has demonstrated the best performance among all reported sulfur/MOFs composite cathode materials, with a reversible capacity of about 700 mAh/g at 0.5 C after 200 cycles. PMID:26323942

  1. Peptide Synthesis through Cell-Free Expression of Fusion Proteins Incorporating Modified Amino Acids as Latent Cleavage Sites for Peptide Release.

    PubMed

    Liutkus, Mantas; Fraser, Samuel A; Caron, Karine; Stigers, Dannon J; Easton, Christopher J

    2016-05-17

    Chlorinated analogues of Leu and Ile are incorporated during cell-free expression of peptides fused to protein, by exploiting the promiscuity of the natural biosynthetic machinery. They then act as sites for clean and efficient release of the peptides simply by brief heat treatment. Dehydro analogues of Leu and Ile are similarly incorporated as latent sites for peptide release through treatment with iodine under cold conditions. These protocols complement enzyme-catalyzed methods and have been used to prepare calcitonin, gastrin-releasing peptide, cholecystokinin-7, and prolactin-releasing peptide prohormones, as well as analogues substituted with unusual amino acids, thus illustrating their practical utility as alternatives to more traditional chemical peptide synthesis. PMID:26918308

  2. Effect of amino acid substitutions in a potential metal-binding site of AnfA on expression from the anfH promoter in Azotobacter vinelandii.

    PubMed

    Premakumar, R; Loveless, T M; Bishop, P E

    1994-10-01

    AnfA, an activator required for transcription of the structural genes encoding nitrogenase 3 (anfHDGK) in Azotobacter vinelandii, has a potential metal-binding site [(S19)H(C21)FTGE(C26)R] in its N terminus. Growth studies and expression of an anfH-lacZ fusion in mutants containing amino acid substitutions in this site indicate that Ser-19 is not required for AnfA activity whereas Cys-21 and Cys-26 are required. Residual expression of the anfH-lacZ fusion in AnfA- mutants was found to be due to activation by VnfA, the activator required for expression of genes encoding nitrogenase 2.

  3. Assessment of acid rock drainage pollutants release in the uranium mining site of Poços de Caldas--Brazil.

    PubMed

    Fernandes, H M; Franklin, M R

    2001-01-01

    We compared three different techniques to assess acid drainage occurrence connected to pyritic waste rock piles at a uranium mining and milling site in Poços de Caldas--Brazil: (1) mass balance calculations, (2) column leaching experiments and (3) geochemical modelling. The study site was chosen because all the drainage coming from the pile is collected in one holding pond and a huge database (monitoring program) was available. The three independent methods predicted similar values for the intrinsic oxidation rate (IOR) (about 10(-9) kg m-3 s-1). We estimate the total time for consumption of all oxidizable material in the dump to be greater than 500 years. Geochemical model results showed a good agreement between predicted sulphate concentrations in relation to those found in the waste pile drainage, although the Al values were overestimated and pH values were underestimated. PMID:11379073

  4. Denuder/filter sampling of organic acids and organosulfates at urban and boreal forest sites: Gas/particle distribution and possible sampling artifacts

    NASA Astrophysics Data System (ADS)

    Kristensen, Kasper; Bilde, Merete; Aalto, Pasi P.; Petäjä, Tuukka; Glasius, Marianne

    2016-04-01

    Carboxylic acids and organosulfates comprise an important fraction of atmospheric secondary organic aerosols formed from both anthropogenic and biogenic precursors. The partitioning of these compounds between the gas and particle phase is still unclear and further research is warranted to better understand the abundance and effect of organic acids and organosulfates on the formation and properties of atmospheric aerosols. This work compares atmospheric aerosols collected at an urban and a boreal forest site using two side-by-side sampling systems; a high volume sampler (HVS) and a low volume (LVS) denuder/filter sampling system allowing for separate collection of gas- and particle-phase organics. All particle filters and denuder samples were collected at H.C. Andersen Boulevard (HCAB), Copenhagen, Denmark in the summer of 2010, and at the remote boreal forest site at Hyytiälä forestry field station in Finland in the summer of 2012. The chemical composition of gas- and particle-phase secondary organic aerosol was investigated by ultra-high performance liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC/ESI-Q-TOFMS), with a focus on carboxylic acids and organosulfates. Results show gas-phase concentrations higher than those observed in the particle phase by a factor of 5-6 in HCAB 2010 and 50-80 in Hyytiälä 2012. Although abundant in the particle phase, no organosulfates were detected in the gas phase at either site. Through a comparison of samples collected by the HVS and the LVS denuder/filter sampling system we evaluate the potential artifacts associated with sampling of atmospheric aerosols. Such comparison shows that particle phase concentrations of semi-volatile organic acids obtained from the filters collected by HVS are more than two times higher than concentrations obtained from filters collected using LVS denuder/filter system. In most cases, higher concentrations of organosulfates are observed in particles

  5. Structure of a highly acidic β-lactamase from the moderate halophile Chromohalobacter sp. 560 and the discovery of a Cs{sup +}-selective binding site

    SciTech Connect

    Arai, Shigeki; Yonezawa, Yasushi; Okazaki, Nobuo; Matsumoto, Fumiko; Shibazaki, Chie; Shimizu, Rumi; Yamada, Mitsugu; Adachi, Motoyasu; Tamada, Taro; Kawamoto, Masahide; Tokunaga, Hiroko; Ishibashi, Matsujiro; Blaber, Michael; Tokunaga, Masao; Kuroki, Ryota

    2015-03-01

    The tertiary structure of a β-lactamase derived from the halobacterium Chromohalobacter sp. 560 (HaBLA) was determined by X-ray crystallography. Three unique Sr{sup 2+}-binding sites and one Cs{sup +}-binding site were discovered in the HaBLA molecule. Environmentally friendly absorbents are needed for Sr{sup 2+} and Cs{sup +}, as the removal of the radioactive Sr{sup 2+} and Cs{sup +} that has leaked from the Fukushima Nuclear Power Plant is one of the most important problems in Japan. Halophilic proteins are known to have many acidic residues on their surface that can provide specific binding sites for metal ions such as Cs{sup +} or Sr{sup 2+}. The crystal structure of a halophilic β-lactamase from Chromohalobacter sp. 560 (HaBLA) was determined to resolutions of between 1.8 and 2.9 Å in space group P3{sub 1} using X-ray crystallography. Moreover, the locations of bound Sr{sup 2+} and Cs{sup +} ions were identified by anomalous X-ray diffraction. The location of one Cs{sup +}-specific binding site was identified in HaBLA even in the presence of a ninefold molar excess of Na{sup +} (90 mM Na{sup +}/10 mM Cs{sup +}). From an activity assay using isothermal titration calorimetry, the bound Sr{sup 2+} and Cs{sup +} ions do not significantly affect the enzymatic function of HaBLA. The observation of a selective and high-affinity Cs{sup +}-binding site provides important information that is useful for the design of artificial Cs{sup +}-binding sites that may be useful in the bioremediation of radioactive isotopes.