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Sample records for surface trp residue

  1. Förster energy-transfer studies between Trp residues of alpha1-acid glycoprotein (orosomucoid) and the glycosylation site of the protein.

    Science.gov (United States)

    Albani, Jihad R

    2003-10-10

    Energy-transfer studies between Trp residues of alpha(1)-acid glycoprotein and the fluorescent probe Calcofluor White were performed. Calcofluor White interacts with carbohydrate residues of the protein, while the three Trp residues are located at the surface (Trp-160) and in hydrophobic domains of the protein (Trp-25 and Trp-122). Binding of Calcofluor to the protein induces a decrease in the fluorescence intensity of the Trp residues accompanied by an increase of that of Calcofluor White. Efficiency (E) of Trp fluorescence quenching was determined to be equal to 45%, and the Förster distance R(o), at which the efficiency of energy transfer is 50%, was calculated to be 18.13 A. This low distance and the value of the efficiency clearly indicate that energy transfer between Trp residues and Calcofluor White is weak.

  2. Effect of the secondary structure of carbohydrate residues of alpha 1-acid glycoprotein (orosomucoid) on the local dynamics of Trp residues.

    Science.gov (United States)

    Albani, Jihad René

    2004-01-01

    We studied in this work the relation between the secondary structure of the carbohydrate residues of alpha1-acid glycoprotein and the local motions of Trp residues of the protein. We measured for this purpose the fluorescence emission intensity and anisotropy of the Trp residues between -46 and +30 degrees of the sialylated and asialylated protein. Our results indicate that, in both forms, the global profile of the emission intensity with temperature shows that Trp residues display static and collisional interaction with the neighboring amino acids. However, the profile of the asialylated form is more structured than that observed for the sialylated protein. The Y-plot analysis of the emission-anisotropy results indicated that the frictional resistance to rotation of the surface Trp residue is less important in the sialylated protein than in the asialylated form. This result is in good agreement with the fact that, in the asialylated conformation, the carbohydrate residues are closer to the protein surface than in the sialylated form, thereby increasing the contact of the surface Trp residue with the neighboring amino acids. Also, the interaction between the carbohydrate residues and the surface Trp residue contributes to the modification of the frictional resistance to rotation of the fluorophore.

  3. Fluorescence properties of porcine odorant binding protein Trp 16 residue

    Energy Technology Data Exchange (ETDEWEB)

    Albani, Jihad Rene, E-mail: Jihad-Rene.Albani@univ-lille1.f [Laboratoire de Biophysique Moleculaire, Universite des Sciences et Technologies de Lille, F-59655 Villeneuve d' Ascq Cedex (France)

    2010-11-15

    Summary: The present work deals with fluorescence studies of adult porcine odorant binding protein at pH=7.5. At this pH, the protein is a dimer, each monomer contains one tryptophan residue. Our results show that tryptophan residue displays significant motions and emits with three fluorescence lifetimes. Decay associated spectra showed that the three lifetime's components emanate from sub-structures surrounded by the same microenvironment.

  4. A potential yeast actin allosteric conduit dependent on hydrophobic core residues val-76 and trp-79.

    Science.gov (United States)

    Wen, Kuo-Kuang; McKane, Melissa; Stokasimov, Ema; Fields, Jonathon; Rubenstein, Peter A

    2010-07-02

    Intramolecular allosteric interactions responsible for actin conformational regulation are largely unknown. Previous work demonstrated that replacing yeast actin Val-76 with muscle actin Ile caused decreased nucleotide exchange. Residue 76 abuts Trp-79 in a six-residue linear array beginning with Lys-118 on the surface and ending with His-73 in the nucleotide cleft. To test if altering the degree of packing of these two residues would affect actin dynamics, we constructed V76I, W79F, and W79Y single mutants as well as the Ile-76/Phe-79 and Ile-76/Tyr-79 double mutants. Tyr or Phe should decrease crowding and increase protein flexibility. Subsequent introduction of Ile should restore packing and dampen changes. All mutants showed decreased growth in liquid medium. W79Y alone was severely osmosensitive and exhibited vacuole abnormalities. Both properties were rescued by Ile-76. Phe-79 or Tyr decreased the thermostability of actin and increased its nucleotide exchange rate. These effects, generally greater for Tyr than for Phe, were reversed by introduction of Ile-76. HD exchange showed that the mutations caused propagated conformational changes to all four subdomains. Based on results from phosphate release and light-scattering assays, single mutations affected polymerization in the order of Ile, Phe, and Tyr from least to most. Introduction of Ile-76 partially rescued the polymerization defects caused by either Tyr-79 or Phe-79. Thus, alterations in crowding of the 76-79 residue pair can strongly affect actin conformation and behavior, and these results support the theory that the amino acid array in which they are located may play a central role in actin regulation.

  5. Position-Dependent Influence of the Three Trp Residues on the Membrane Activity of the Antimicrobial Peptide, Tritrpticin

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    Mauricio Arias

    2014-11-01

    Full Text Available Antimicrobial peptides (AMPs constitute promising candidates for the development of new antibiotics. Among the ever-expanding family of AMPs, tritrpticin has strong antimicrobial activity against a broad range of pathogens. This 13-residue peptide has an unusual amino acid sequence that is almost symmetrical and features three central Trp residues with two Arg residues near each end of the peptide. In this work, the role of the three sequential Trp residues in tritrpticin was studied in a systematic fashion by making a series of synthetic peptides with single-, double- and triple-Trp substitutions to Tyr or Ala. 1H NMR and fluorescence spectroscopy demonstrated the ability of all of the tritrpticin-analog peptides to interact with negatively-charged membranes. Consequently, most tritrpticin analogs exhibited the ability to permeabilize synthetic ePC:ePG (egg-yolk phosphatidylcholine (ePC, egg-yolk phosphatidylglycerol (ePG vesicles and live Escherichia coli bacteria. The membrane perturbation characteristics were highly dependent on the location of the Trp residue substitution, with Trp6 being the most important residue and Trp8 the least. The membrane permeabilization activity of the peptides in synthetic and biological membranes was directly correlated with the antimicrobial potency of the peptides against E. coli. These results contribute to the understanding of the role of each of the three Trp residues to the antimicrobial activity of tritrpticin.

  6. Comparisons of Interfacial Phe, Tyr, and Trp Residues as Determinants of Orientation and Dynamics for GWALP Transmembrane Peptides

    OpenAIRE

    Sparks, Kelsey A.; Gleason, Nicholas J.; Gist, Renetra; Langston, Rebekah; Greathouse, Denise V.; Koeppe, Roger E.

    2014-01-01

    Aromatic amino acids often flank the transmembrane alpha helices of integral membrane proteins. By favoring locations within the membrane–water interface of the lipid bilayer, aromatic residues Trp, Tyr, and sometimes Phe may serve as anchors to help stabilize a transmembrane orientation. In this work, we compare the influence of interfacial Trp, Tyr, or Phe residues upon the properties of tilted helical transmembrane peptides. For such comparisons, it has been critical to start with no more ...

  7. Role of Non-Active-Site Residue Trp-93 in the Function and Stability of New Delhi Metallo-β-Lactamase 1

    Science.gov (United States)

    Rehman, M. Tabish

    2015-01-01

    New Delhi metallo-β-lactamase-1 (NDM-1) is expressed by various members of Enterobacteriaceae as a defense mechanism to hydrolyze β-lactam antibiotics. Despite various studies showing the significance of active-site residues in the catalytic mechanism, there is a paucity of reports addressing the role of non-active-site residues in the structure and function of NDM-1. In this study, we investigated the significance of non-active-site residue Trp-93 in the structure and function of NDM-1. We cloned blaNDM-1 from an Enterobacter cloacae clinical strain (EC-15) and introduced the mutation of Trp-93 to Ala (yielding the Trp93Ala mutant) by PCR-based site-directed mutagenesis. Proteins were expressed and purified to homogeneity by affinity chromatography. The MICs of the Trp93Ala mutant were reduced 4- to 8-fold for ampicillin, cefotaxime, ceftazidime, cefoxitin, imipenem, and meropenem. The poor hydrolytic activity of the Trp93Ala mutant was also reflected by its reduced catalytic efficiency. The overall catalytic efficiency of the Trp93Ala mutant was reduced by 40 to 55% (the Km was reduced, while the kcat was similar to that of wild-type NDM-1 [wtNDM-1]). Heat-induced denaturation showed that the ΔGDo and Tm of Trp93Ala mutant were reduced by 1.8 kcal/mol and 4.8°C, respectively. Far-UV circular dichroism (CD) analysis showed that the α-helical content of the Trp93Ala mutant was reduced by 2.9%. The decrease in stability and catalytic efficiency of the Trp93Ala mutant was due to the loss of two hydrogen bonds with Ser-63 and Val-73 and hydrophobic interactions with Leu-65, Val-73, Gln-123, and Asp-124. The study provided insight into the role of non-active-site amino acid residues in the hydrolytic mechanism of NDM-1. PMID:26525789

  8. Identification of a Residue (Glu60) in TRAP Required for Inducing Efficient Transcription Termination at the trp Attenuator Independent of Binding Tryptophan and RNA.

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    McAdams, Natalie M; Patterson, Andrea; Gollnick, Paul

    2017-03-15

    Transcription of the tryptophan (trp) operon in Bacillus subtilis is regulated by an attenuation mechanism. Attenuation is controlled by the trpRNA-binding attenuation protein (TRAP). TRAP binds to a site in the 5' leader region of the nascent trp transcript in response to the presence of excess intracellular tryptophan. This binding induces transcription termination upstream of the structural genes of the operon. In prior attenuation models, the role of TRAP was only to alter the secondary structure of the leader region RNA so as to promote formation of the trp attenuator, which was presumed to function as an intrinsic terminator. However, formation of the attenuator alone has been shown to be insufficient to induce efficient termination, indicating that TRAP plays an additional role in this process. To further examine the function of TRAP, we performed a genetic selection for mutant TRAPs that bind tryptophan and RNA but show diminished termination at the trp attenuator. Five such TRAP mutants were obtained. Four of these have substitutions at Glu60, three of which are Lys (E60K) substitutions and the fourth of which is a Val (E60V) substitution. The fifth mutant obtained contains a substitution at Ile63, which is on the same β-strand of TRAP as Glu60. Purified E60K TRAP binds tryptophan and RNA with properties similar to those of the wild type but is defective at inducing termination at the trp attenuator in vitroIMPORTANCE Prior models for attenuation control of the B. subtilis trp operon suggested that the only role for TRAP is to bind to the leader region RNA and alter its folding to induce formation of an intrinsic terminator. However, several recent studies suggested that TRAP plays an additional role in the termination mechanism. We hypothesized that this function could involve residues in TRAP other than those required to bind tryptophan and RNA. Here we obtained TRAP mutants with alterations at Glu60 that are deficient at inducing termination in the

  9. Functional analysis of three amino acid residues of purR re-pressor, Trp147, Gln-218 and Gln-292 in Salmonella typhi-murium

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The amber mutation sites of 6 purR(am) mutants were determined bycloning and DNA sequencing. The results showed that the mutations were distributed at three different sites in PurR coding region, G721(→A), C933(→T) and C1155(→T), which respectively turn Trp-147,Gln-218 and Gln-292 of PurR into TAG terminal codon. To determine the effect of the three amino acid residues on regulatory function of PurR protein 5 different kinds of tRNA suppressor genes, Su3, Su4, Su6, Su7 and Su9 were used for creating the PurR protein variants with single amino acid substitution. The results indicated that Cys, Glu, Gly, His and Arg which substituted Trp-147 respectively all could not recover the regulation function of PurR. It confirmed that Trp-147 is a critical amino acid for the PurR function. Gln-292 substituted respectively by the same amino acids also could not recover the PurR function, demonstrating that Gln-292 is also an important amino acid residue in PurR.

  10. Structural analysis of a penicillin V acylase from Pectobacterium atrosepticum confirms the importance of two Trp residues for activity and specificity.

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    Avinash, Vellore Sunder; Panigrahi, Priyabrata; Chand, Deepak; Pundle, Archana; Suresh, Cheravakattu Gopalan; Ramasamy, Sureshkumar

    2016-02-01

    Penicillin V acylases (PVA) catalyze the deacylation of the beta-lactam antibiotic phenoxymethylpenicillin (Pen V). They are members of the Ntn hydrolase family and possess an N-terminal cysteine as the main catalytic nucleophile residue. They form the evolutionarily related cholylglycine hydrolase (CGH) group which includes bile salt hydrolases (BSH) responsible for bile deconjugation. Even though a few PVA and BSH structures have been reported, no structure of a functional PVA from Gram-negative bacteria is available. Here, we report the crystal structure of a highly active PVA from Gram-negative Pectobacterium atrosepticum (PaPVA) at 2.5Å resolution. Structural comparison with PVAs from Gram-positive bacteria revealed that PaPVA had a distinctive tetrameric structure and active site organization. In addition, mutagenesis of key active site residues and biochemical characterization of the resultant variants elucidated the role of these residues in substrate binding and catalysis. The importance of residue Trp23 and Trp87 side chains in binding and correct positioning of Pen V by PVAs was confirmed using mutagenesis and substrate docking with a 15ns molecular dynamics simulation. These results establish the unique nature of Gram-negative CGHs and necessitate further research about their substrate spectrum.

  11. Formation of truncated peptide by-products via sequence-specific formyl group transfer from Trp(For) residues to Nα in the course of Boc-SPPS.

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    Azev, Viatcheslav N; Mustaeva, Leila G; Gorbunova, Elena Yu; Molchanov, Maksim V; Rodionov, Igor L

    2013-10-01

    (N(In))-Formyl protective group of tryptophan has been introduced as a base/nucleophile-labile protective group. It has long been known that a free Nα-amino group of the peptide can serve as a nucleophile: an irreversible formyl N(In)  → NH(2) transfer is consistently observed when deformylation is performed last on an otherwise deprotected peptide that possesses free Nα-amino group. Obviously, this particular side reaction should be expected any time free amino group is exposed to Trp(For), but, at the best of our knowledge, has never been reported in the course of Boc-SPPS. In the present communication, we describe a set of appropriately designed model experiments that permitted to detect the title side reaction both in solution and in solid-phase reactions. We observed intermolecular formyl group transfer with a model compound, Trp(For)-NH(2). Importantly, we also observed this migration on solid support with the rate roughly estimated to be up to 1% of residues per minute. We also observed that the formyl-group transfer reaction occurred in a sequence-dependent manner and was suppressed to a non-detectable level using 'in situ neutralization' technique. Because this side reaction is sequence dependent, there might be situations when the rate of the formation of Nα -formyl termination by-products is significant. In other cases, the Nα -For truncated by-products would not contaminate the final peptide significantly but still could be a source of microheterogeneity. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd.

  12. Both the dimerization and immunochemical properties of E-cadherin EC1 domain depend on Trp(156) residue

    DEFF Research Database (Denmark)

    Laur, Oscar Y; Klingelhöfer, Jörg; Troyanovsky, Regina B;

    2002-01-01

    Using site-directed mutagenesis, we show in this paper that the adhesive interface detected in cadherin crystals is unlikely to mediate adhesive interaction between myc- and flag-tagged E-cadherin molecules in human A-431 cells. We also found that a critical residue within this interface, His(233...

  13. Specific function of the Met-Tyr-Trp adduct radical and residues Arg-418 and Asp-137 in the atypical catalase reaction of catalase-peroxidase KatG.

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    Zhao, Xiangbo; Khajo, Abdelahad; Jarrett, Sanchez; Suarez, Javier; Levitsky, Yan; Burger, Richard M; Jarzecki, Andrzej A; Magliozzo, Richard S

    2012-10-26

    Catalase activity of the dual-function heme enzyme catalase-peroxidase (KatG) depends on several structural elements, including a unique adduct formed from covalently linked side chains of three conserved amino acids (Met-255, Tyr-229, and Trp-107, Mycobacterium tuberculosis KatG numbering) (MYW). Mutagenesis, electron paramagnetic resonance, and optical stopped-flow experiments, along with calculations using density functional theory (DFT) methods revealed the basis of the requirement for a radical on the MYW-adduct, for oxyferrous heme, and for conserved residues Arg-418 and Asp-137 in the rapid catalase reaction. The participation of an oxyferrous heme intermediate (dioxyheme) throughout the pH range of catalase activity is suggested from our finding that carbon monoxide inhibits the activity at both acidic and alkaline pH. In the presence of H(2)O(2), the MYW-adduct radical is formed normally in KatG[D137S] but this mutant is defective in forming dioxyheme and lacks catalase activity. KatG[R418L] is also catalase deficient but exhibits normal formation of the adduct radical and dioxyheme. Both mutants exhibit a coincidence between MYW-adduct radical persistence and H(2)O(2) consumption as a function of time, and enhanced subunit oligomerization during turnover, suggesting that the two mutations disrupting catalase turnover allow increased migration of the MYW-adduct radical to protein surface residues. DFT calculations showed that an interaction between the side chain of residue Arg-418 and Tyr-229 in the MYW-adduct radical favors reaction of the radical with the adjacent dioxyheme intermediate present throughout turnover in WT KatG. Release of molecular oxygen and regeneration of resting enzyme are thereby catalyzed in the last step of a proposed catalase reaction.

  14. Fluorescence kinetics of Trp-Trp dipeptide and its derivatives in water via ultrafast fluorescence spectroscopy.

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    Jia, Menghui; Yi, Hua; Chang, Mengfang; Cao, Xiaodan; Li, Lei; Zhou, Zhongneng; Pan, Haifeng; Chen, Yan; Zhang, Sanjun; Xu, Jianhua

    2015-08-01

    Ultrafast fluorescence dynamics of Tryptophan-Tryptophan (Trp-Trp/Trp2) dipeptide and its derivatives in water have been investigated using a picosecond resolved time correlated single photon counting (TCSPC) apparatus together with a femtosecond resolved upconversion spectrophotofluorometer. The fluorescence decay profiles at multiple wavelengths were fitted by a global analysis technique. Nanosecond fluorescence kinetics of Trp2, N-tert-butyl carbonyl oxygen-N'-aldehyde group-l-tryptophan-l-tryptophan (NBTrp2), l-tryptophan-l-tryptophan methyl ester (Trp2Me), and N-acetyl-l-tryptophan-l-tryptophan methyl ester (NATrp2Me) exhibit multi-exponential decays with the average lifetimes of 1.99, 3.04, 0.72 and 1.22ns, respectively. Due to the intramolecular interaction between two Trp residues, the "water relaxation" lifetime was observed around 4ps, and it is noticed that Trp2 and its derivatives also exhibit a new decay with a lifetime of ∼100ps, while single-Trp fluorescence decay in dipeptides/proteins shows 20-30ps. The intramolecular interaction lifetime constants of Trp2, NBTrp2, Trp2Me and NATrp2Me were then calculated to be 3.64, 0.93, 11.52 and 2.40ns, respectively. Candidate mechanisms (including heterogeneity, solvent relaxation, quasi static self-quenching or ET/PT quenching) have been discussed.

  15. Spectroscopic evidence for an engineered, catalytically active Trp radical that creates the unique reactivity of lignin peroxidase.

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    Smith, Andrew T; Doyle, Wendy A; Dorlet, Pierre; Ivancich, Anabella

    2009-09-22

    The surface oxidation site (Trp-171) in lignin peroxidase (LiP) required for the reaction with veratryl alcohol a high-redox-potential (1.4 V) substrate, was engineered into Coprinus cinereus peroxidase (CiP) by introducing a Trp residue into a heme peroxidase that has similar protein fold but lacks this activity. To create the catalytic activity toward veratryl alcohol in CiP, it was necessary to reproduce the Trp site and its negatively charged microenvironment by means of a triple mutation. The resulting D179W+R258E+R272D variant was characterized by multifrequency EPR spectroscopy. The spectra unequivocally showed that a new Trp radical [g values of g(x) = 2.0035(5), g(y) = 2.0027(5), and g(z) = 2.0022(1)] was formed after the [Fe(IV)=O Por(*+)] intermediate, as a result of intramolecular electron transfer between Trp-179 and the porphyrin. Also, the EPR characterization crucially showed that [Fe(IV)=O Trp-179(*)] was the reactive intermediate with veratryl alcohol. Accordingly, our work shows that it is necessary to take into account the physicochemical properties of the radical, fine-tuned by the microenvironment, as well as those of the preceding [Fe(IV)=O Por(*+)] intermediate to engineer a catalytically competent Trp site for a given substrate. Manipulation of the microenvironment of the Trp-171 site in LiP allowed the detection by EPR spectroscopy of the Trp-171(*), for which direct evidence has been missing so far. Our work also highlights the role of Trp residues as tunable redox-active cofactors for enzyme catalysis in the context of peroxidases with a unique reactivity toward recalcitrant substrates that require oxidation potentials not realized at the heme site.

  16. Functional diversity and evolutionary dynamics of thermoTRP channels.

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    Saito, Shigeru; Tominaga, Makoto

    2015-03-01

    Animals have evolved sophisticated physiological systems for sensing ambient temperature since changes in environmental temperatures affect various biological processes. Thermosensitive transient receptor potential (thermoTRP) channels serve as thermal sensors in diverse animal species. They are multimodal receptors that are activated by temperature as well as other physical and chemical stimuli. Since thermoTRP channels are calcium permeable non-selective cation channels, their activation leads to an influx of calcium and sodium ions into the cell and triggers downstream signal transduction. ThermoTRP channels have been characterized in diverse animal species over the past several years, illuminating the diversification of thermoTRP channels in the course of evolution. The gene repertoires of thermoTRP channels differ among animal species. Additionally, in some cases, the temperature and chemical sensitivities among orthologous thermoTRP channels vary among species. The evolutionary flexibility of thermoTRP channels enabled them to contribute to unique physiological systems such as infrared sensation in snakes and bats and seasonal adaptation in silk moth. On the other hand, the functional differences of thermoTRP channels among species have been utilized for understanding the molecular basis for their activation (or inhibition) mechanisms, and amino acid residues (or domains) responsible for the respective channel properties have been identified in various thermoTRP channels. Here we summarize the current understanding of the functional diversity and evolutionary dynamics of thermoTRP channels.

  17. TRP channels in schistosomes

    Directory of Open Access Journals (Sweden)

    Swarna Bais

    2016-12-01

    Full Text Available Praziquantel (PZQ is effectively the only drug currently available for treatment and control of schistosomiasis, a disease affecting hundreds of millions of people worldwide. Many anthelmintics, likely including PZQ, target ion channels, membrane protein complexes essential for normal functioning of the neuromusculature and other tissues. Despite this fact, only a few classes of parasitic helminth ion channels have been assessed for their pharmacological properties or for their roles in parasite physiology. One such overlooked group of ion channels is the transient receptor potential (TRP channel superfamily. TRP channels share a common core structure, but are widely diverse in their activation mechanisms and ion selectivity. They are critical to transducing sensory signals, responding to a wide range of external stimuli. They are also involved in other functions, such as regulating intracellular calcium and organellar ion homeostasis and trafficking. Here, we review current literature on parasitic helminth TRP channels, focusing on those in schistosomes. We discuss the likely roles of these channels in sensory and locomotor activity, including the possible significance of a class of TRP channels (TRPV that is absent in schistosomes. We also focus on evidence indicating that at least one schistosome TRP channel (SmTRPA has atypical, TRPV1-like pharmacological sensitivities that could potentially be exploited for future therapeutic targeting.

  18. Post-Translational Modifications of TRP Channels

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    Olaf Voolstra

    2014-04-01

    Full Text Available Transient receptor potential (TRP channels constitute an ancient family of cation channels that have been found in many eukaryotic organisms from yeast to human. TRP channels exert a multitude of physiological functions ranging from Ca2+ homeostasis in the kidney to pain reception and vision. These channels are activated by a wide range of stimuli and undergo covalent post-translational modifications that affect and modulate their subcellular targeting, their biophysical properties, or channel gating. These modifications include N-linked glycosylation, protein phosphorylation, and covalent attachment of chemicals that reversibly bind to specific cysteine residues. The latter modification represents an unusual activation mechanism of ligand-gated ion channels that is in contrast to the lock-and-key paradigm of receptor activation by its agonists. In this review, we summarize the post-translational modifications identified on TRP channels and, when available, explain their physiological role.

  19. TRP channels: an overview

    DEFF Research Database (Denmark)

    Pedersen, Stine Falsig; Owsianik, Grzegorz; Nilius, Bernd

    2005-01-01

    to a plethora of data on the roles of TRPs in a variety of tissues and species, including mammals, insects, and yeast. The present review summarizes the most pertinent recent evidence regarding the structural and functional properties of TRP channels, focusing on the regulation and physiology of mammalian TRPs....

  20. Role of the middle residue in the triple tryptophan electron transfer chain of DNA photolyase: ultrafast spectroscopy of a Trp-->Phe mutant.

    Science.gov (United States)

    Lukacs, Andras; Eker, André P M; Byrdin, Martin; Villette, Sandrine; Pan, Jie; Brettel, Klaus; Vos, Marten H

    2006-08-17

    Photoreduction of the semi-reduced flavin adenine dinucleotide cofactor FADH* in DNA photolyase from Escherichia coli into FADH- involves three tryptophan (W) residues that form a closely spaced electron-transfer chain FADH*-W382-W359-W306. To investigate this process, we have constructed a mutant photolyase in which W359 is replaced by phenylalanine (F). Monitoring its photoproducts by femtosecond spectroscopy, the excited-state FADH* was found to decay in approximately 30 ps, similar as in wild type (WT) photolyase. In contrast to WT, however, in W359F mutant photolyase the ground-state FADH* fully recovered virtually concomitantly with the decay of its excited state and, despite the presence of the primary electron donor W382, no measurable flavin reduction was observed at any time. Thus, W359F photolyase appears to behave like many other flavoproteins, where flavin excited states are quenched by very short-lived oxidation of aromatic residues. Our analysis indicates that both charge recombination of the primary charge separation state FADH-W382*+ and (in WT) electron transfer from W359 to W382*+ occur with time constants FADH* electron-transfer step. Our results provide a first experimental indication that electron transfer between aromatic residues can take place on the time scale of approximately 10(-12) s.

  1. Drosophila nociceptors mediate larval aversion to dry surface environments utilizing both the painless TRP channel and the DEG/ENaC subunit, PPK1.

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    Wayne A Johnson

    Full Text Available A subset of sensory neurons embedded within the Drosophila larval body wall have been characterized as high-threshold polymodal nociceptors capable of responding to noxious heat and noxious mechanical stimulation. They are also sensitized by UV-induced tissue damage leading to both thermal hyperalgesia and allodynia very similar to that observed in vertebrate nociceptors. We show that the class IV multiple-dendritic(mdIV nociceptors are also required for a normal larval aversion to locomotion on to a dry surface environment. Drosophila melanogaster larvae are acutely susceptible to desiccation displaying a strong aversion to locomotion on dry surfaces severely limiting the distance of movement away from a moist food source. Transgenic inactivation of mdIV nociceptor neurons resulted in larvae moving inappropriately into regions of low humidity at the top of the vial reflected as an increased overall pupation height and larval desiccation. This larval lethal desiccation phenotype was not observed in wild-type controls and was completely suppressed by growth in conditions of high humidity. Transgenic hyperactivation of mdIV nociceptors caused a reciprocal hypersensitivity to dry surfaces resulting in drastically decreased pupation height but did not induce the writhing nocifensive response previously associated with mdIV nociceptor activation by noxious heat or harsh mechanical stimuli. Larvae carrying mutations in either the Drosophila TRP channel, Painless, or the degenerin/epithelial sodium channel subunit Pickpocket1(PPK1, both expressed in mdIV nociceptors, showed the same inappropriate increased pupation height and lethal desiccation observed with mdIV nociceptor inactivation. Larval aversion to dry surfaces appears to utilize the same or overlapping sensory transduction pathways activated by noxious heat and harsh mechanical stimulation but with strikingly different sensitivities and disparate physiological responses.

  2. TRP channels and pain.

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    Julius, David

    2013-01-01

    Nociception is the process whereby primary afferent nerve fibers of the somatosensory system detect noxious stimuli. Pungent irritants from pepper, mint, and mustard plants have served as powerful pharmacological tools for identifying molecules and mechanisms underlying this initial step of pain sensation. These natural products have revealed three members of the transient receptor potential (TRP) ion channel family--TRPV1, TRPM8, and TRPA1--as molecular detectors of thermal and chemical stimuli that activate sensory neurons to produce acute or persistent pain. Analysis of TRP channel function and expression has validated the existence of nociceptors as a specialized group of somatosensory neurons devoted to the detection of noxious stimuli. These studies are also providing insight into the coding logic of nociception and how specification of nociceptor subtypes underlies behavioral discrimination of noxious thermal, chemical, and mechanical stimuli. Biophysical and pharmacological characterization of these channels has provided the intellectual and technical foundation for developing new classes of analgesic drugs.

  3. Residual stress around the cortical surface in bovine femoral diaphysis.

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    Yamada, Satoshi; Tadano, Shigeru

    2010-04-01

    Residual stress in living tissue plays an important role in mechanical strength. We have reported that residual stress exists in the bone tissue of a rabbit's tibiofibula. The purpose of this study is to measure the residual stress around the outer cortical region of bovine femoral diaphysis and to discuss the distribution of the stress. This work proposed the sin(2) psi method of X-ray diffraction to the measurement of residual stresses in bone tissue. In this method, residual stress can be estimated from the variation in the interplanar spacings orientated to a number of directions without the lattice strain in the stress direction. Four-point bending tests of strip specimens taken from bovine femoral diaphysis were carried out during X-ray irradiation in advance. In the proximal, middle, and distal sections of bovine femoral diaphyses, the residual stresses at the cortical surface were measured using characteristic Mo-Kalpha X-rays. The bending tests of strip specimens with X-ray irradiation showed that the method could reliably estimate residual stresses in the bone tissue. The residual stress of the bone axial direction was larger than that of the circumferential direction. The stresses in the middle part of five diaphyses along the bone axial direction were tensile. The maximum stress was 162 MPa at the lateral position and the minimum was 78 MPa at the posterior position. The residual stress in the bone axial direction varies around the circumferential region. In addition, the bone axial distributions of residual stresses were different in the proximal, middle, and distal sections of the individual femur. Furthermore, it was confirmed that residual stress in the bone tissue was released by the cutting out of the specimen. The residual stresses in bone tissue could be measured by this method. The results show that residual stress in the bone axial direction at the cortical surface in bovine femoral diaphysis is tensile and varies around the circumferential

  4. Aging of residual surface resistance of superconducting lead cavities

    DEFF Research Database (Denmark)

    Danielsen, M.

    1972-01-01

    Measurements of the residual surface resistance of superconducting lead cavities as a function of time during a period of a month showed an oscillating variation. An explanation of the ageing curves is proposed. ©1972 The American Institute of Physics......Measurements of the residual surface resistance of superconducting lead cavities as a function of time during a period of a month showed an oscillating variation. An explanation of the ageing curves is proposed. ©1972 The American Institute of Physics...

  5. Functional analysis of three amino acid residues of purR re-pressor, Trp147, Gln-218 and Gln-292 in Salmonella typhi-murium

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Hesheng

    2001-01-01

    [1]Zalkin, H., Nygaard, P., Escherichia coli and Salmmella typhimurium Cellular and Molecular Biology, Washington D.C: American Society for Microbiology, 1996, 561-579.[2]Maria, A. S., Choi, K. Y., Lu, F. et al., Mechanism of co-repressor mediated specific DNA binding by the purine repressor, Cell, 1995, 83: 147.[3]Choi, K. Y., Fu, L., Zalkin, H., Mutagenesis of amino acid residues required for binding of co-repressors to the purine repressor, J. Biol. Chem., 1993, 269: 24066.[4]Lu, F., Brennan, R. G., Zalkin, H., Escherichia coli purine repressor: key residues for the allsteic transition between active and inactive conformation and for inter domain signaling, Biochemistry, 1998, 37: 15680.[5]Tang Hua, Qin Junchuan, Wang Aoquan, Regulation of purine biosynthetic genes expression in Salmonella typhimurium (VI)-- Isolation and characterization of superrepressor mutants, Chinese Journal of Genetics (in Chinese), 1998, 25(2): 181.[6]Zhang Hesheng, Wang Aoquan, Regulation of purine biosynthetic genes expression in Salmonella typhimurium (X)--Isola-tion of purR(am) mutant and preliminary study of amino acid substitution, Chinese Journal of Genetics (in Chinese), 2000, 27(2): 170.[7]Davis, R. W., Roth, J. R., Advanced Bacteria Genetics, NY: Cold Spring Harbor Laboratory Press, 1984.[8]Miller, J. H., Experiments in Molecular Genetics, NY: Cold Spring Harbor Laboratory Press, 1972.[9]Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning, A Laboratory Manual, NY: Cold Spring Harbor laboratory Press, 1989.[10] Katzif, S. D., Lu, C. D., Abdelal, A.T., Salmonela typhimurium purine nucleotide synthesis repressor (purR), GenBank, Ac-cession AF040636, 1998.[11] Weicket,M.J.,Adhya,S., A family of bacteria regulators homologous to Gal and Lac repressors,J.Biol.Chem.,1992,267:15869.

  6. Chemical Modification and Fluorescence Spectrum of Tryptophan Residues in Pullulanase

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Tryptophan(Trp) residues in a pullulanase were modified by N-bromosuccinimide(NBS). The results of the Spande method indicate that there are 18 Trp residues in the pullulanase and nine of them are located on the surface of the enzyme. Three of these Trp residues are nonessential residues which show the fastest reaction rate according to the Zou′s plot. Two of the seven relative faster reacting residues are essential for the activity of the enzyme. The other eight are the slowest in the reaction rate or non-reactive residues for the reaction. The fluorescence and circular dichroism(CD) spectra of the pullulanase have been changed after the reaction with NBS. Potassium iodide(KI) and acrylamide also have remarkable influences on the fluorescence spectra of the pullulanase.

  7. Killing of Staphylococcus aureus and Salmonella enteritidis and neutralization of lipopolysaccharide by 17-residue bovine lactoferricins: improved activity of Trp/Ala-containing molecules

    Science.gov (United States)

    Hao, Ya; Yang, Na; Wang, Xiumin; Teng, Da; Mao, Ruoyu; Wang, Xiao; Li, Zhanzhan; Wang, Jianhua

    2017-01-01

    Bovine lactoferricin (LfcinB) has potent antibacterial, antifungal and antiparasitic activities but is also hemolytic. Our objective was to identify LfcinB17-31 derivatives with reduced hemolysis and improved antimicrobial activity via substituting Cys3, Arg4, Gln7, Met10, and Gly14 with more hydrophobic residues. Two peptides, Lfcin4 and Lfcin5, showed higher activity against Staphylococcus aureus and Salmonella enteritidis and lower hemolytic activity than the parent peptide LfcinB17-31. These peptides permeabilized the outer and inner membranes of S. enteritidis; however, Lfcin5 did not permeabilize the inner membrane of S. aureus. Gel retardation and circular dichroism spectra showed that Lfcin4 and Lfcin5 bound to bacterial genomic DNA. Lfcin4 inhibited DNA, RNA and protein synthesis. Both peptides induced the peeling of membranes and the lysis of S. enteritidis. At doses of 10 and 15 mg/kg, Lfcin4 and Lfcin5 reduced the bacterial counts in infected thigh muscles by 0.03‒0.10 and 0.05‒0.63 log10 CFU/g of tissue, respectively, within 10 h. Lfcin4 and Lfcin5 enhanced the survival rate of endotoxemic mice; reduced serum IL-6, IL-1β and TNF-α levels; and protected mice from lipopolysaccharide-induced lung injury. These data suggest that Lfcin4 and Lfcin5 may be antimicrobial and anti-endotoxin peptides that could serve as the basis for the development of dual-function agents. PMID:28287172

  8. Engineered Surfaces for Mitigation of Insect Residue Adhesion

    Science.gov (United States)

    Siochi, Emilie J.; Smith, Joseph G.; Wohl, Christopher J.; Gardner, J. M.; Penner, Ronald K.; Connell, John W.

    2013-01-01

    Maintenance of laminar flow under operational flight conditions is being investigated under NASA s Environmentally Responsible Aviation (ERA) Program. Among the challenges with natural laminar flow is the accretion of residues from insect impacts incurred during takeoff or landing. Depending on air speed, temperature, and wing structure, the critical residue height for laminar flow disruption can be as low as 4 microns near the leading edge. In this study, engineered surfaces designed to minimize insect residue adhesion were examined. The coatings studied included chemical compositions containing functional groups typically associated with abhesive (non-stick) surfaces. To reduce surface contact by liquids and enhance abhesion, the engineered surfaces consisted of these coatings doped with particulate additives to generate random surface topography, as well as coatings applied to laser ablated surfaces having precision patterned topographies. Performance evaluation of these surfaces included contact angle goniometry of pristine coatings and profilometry of surfaces after insect impacts were incurred in laboratory scale tests, wind tunnel tests and flight tests. The results illustrate the complexity of designing antifouling surfaces for effective insect contamination mitigation under dynamic conditions and suggest that superhydrophobic surfaces may not be the most effective solution for preventing insect contamination on aircraft wing leading edges.

  9. Assessment of interplay between UV wavelengths, material surfaces and food residues in open surface hygiene validation

    DEFF Research Database (Denmark)

    Abban, Stephen; Jakobsen, Mogens; Jespersen, Lene

    2014-01-01

    The use of UV-visible radiation for detecting invisible residue on different surfaces as a means of validating cleanliness was investigated. Wavelengths at 365, 395, 435, 445, 470 and 490 nm from a monochromator were used to detect residues of beef, chicken, apple, mango and skim milk. These were......-time hygiene validation of surfaces....

  10. INTERFACE DEVICE FOR NONDESTRUCTIVE TESTING OF RESIDUAL SURFACE STRESSES

    Directory of Open Access Journals (Sweden)

    Gennady A. Perepelkin

    2016-01-01

    Full Text Available The paper considers the organization of connection of a personal computer with a device for nondestructive testing of residual surface stresses. The device works is based on the phenomenon of diffraction of ionizing radiation from the crystal lattice near the surface of the crystallites. Proposed software interface to the organization for each type of user: the device developers, administrators, users. Some aspects of the organization of communication microcontroller to a PC via USB-port

  11. Persistence and residue activity of deltamethrin on indoor residual spraying surfaces against malaria vectors in southeastern Iran

    Institute of Scientific and Technical Information of China (English)

    Abtahi Mohammad; Shayeghi Mansoreh; Khoobdel Mehdi; Vatandoost Hasan; Abaei Mohammad Reza; Akbarzadeh Kamran

    2011-01-01

    Objective:To evaluate the efficacy of deltamethrin and find a relation between persistence and residue of this insecticide on the prevalent surfaces against malaria vectors in southeastern Iran. Methods:After indoor residual spraying on prevalent surfaces in studied areas (plaster and mud as absorbent surfaces, wood as non absorbent surface and filter paper as control) for malaria control, conical tests as a bioassay method and chromatographic method as an analytical method were used for evolution of persistence and residue of deltamethrin insecticide. Results were investigated statistically by ANOVA and Tukey-HSD tests for determining relations or differences between residue and persistence of deltamethrin. Results:According to the results, there was no significant difference between mortality rates from bioassay tests on different surfaces, and deltamethrin kept its utility to malaria vector control until 120 days after indoor residual spraying on these surfaces. In the case of residue, there was no significant relation between residue amounts and mortality rates on different surfaces, whereas this relation existed between residual amounts on filter papers and mortality rates from bioassay tests. Conclusions: This study shows that measurement of residue in filter papers is a suitable tool for evolution and dictum of efficiency of deltamethrin insecticide in indoor residual spraying for malaria control.

  12. Prediction and Optimization of Residual Stresses on Machined Surface and Sub-Surface in MQL Turning

    Science.gov (United States)

    Ji, Xia; Zou, Pan; Li, Beizhi; Rajora, Manik; Shao, Yamin; Liang, Steven Y.

    Residual stress in the machined surface and subsurface is affected by materials, machining conditions, and tool geometry and can affect the component life and service quality significantly. Empirical or numerical experiments are commonly used for determining residual stresses but these are very expensive. There has been an increase in the utilization of minimum quantity lubrication (MQL) in recent years in order to reduce the cost and tool/part handling efforts, while its effect on machined part residual stress, although important, has not been explored. This paper presents a hybrid neural network that is trained using Simulated Annealing (SA) and Levenberg-Marquardt Algorithm (LM) in order to predict the values of residual stresses in cutting and radial direction on the surface and within the work piece after the MQL face turning process. Once the ANN has successfully been trained, an optimization procedure, using Genetic Algorithm (GA), is applied in order to find the best cutting conditions in order to minimize the surface tensile residual stresses and maximize the compressive residual stresses within the work piece. The optimization results show that the usage of MQL decreases the surface tensile residual stresses and increases the compressive residual stresses within the work piece.

  13. TRP Channels in Human Prostate

    Directory of Open Access Journals (Sweden)

    Carl Van Haute

    2010-01-01

    Full Text Available This review gives an overview of morphological and functional characteristics in the human prostate. It will focus on the current knowledge about transient receptor potential (TRP channels expressed in the human prostate, and their putative role in normal physiology and prostate carcinogenesis. Controversial data regarding the expression pattern and the potential impact of TRP channels in prostate function, and their involvement in prostate cancer and other prostate diseases, will be discussed.

  14. Molecular cloning of a preprohormone from Hydra magnipapillata containing multiple copies of Hydra-L Wamide (Leu-Trp-NH2) neuropeptides: evidence for processing at Ser and Asn residues

    DEFF Research Database (Denmark)

    Leviev, I; Williamson, M; Grimmelikhuijzen, C J

    1997-01-01

    The simple, freshwater polyp Hydra is often used as a model to study development in cnidarians. Recently, a neuropeptide, < Glu-Gln-Pro-Gly-Leu-Trp-NH2, has been isolated from sea anemones that induces metamorphosis in a hydroid planula larva to become a polyp. Here, we have cloned a preprohormon...

  15. Trafficking of ThermoTRP Channels

    Directory of Open Access Journals (Sweden)

    Clotilde Ferrandiz-Huertas

    2014-08-01

    Full Text Available ThermoTRP channels (thermoTRPs define a subfamily of the transient receptor potential (TRP channels that are activated by changes in the environmental temperature, from noxious cold to injurious heat. Acting as integrators of several stimuli and signalling pathways, dysfunction of these channels contributes to several pathological states. The surface expression of thermoTRPs is controlled by both, the constitutive and regulated vesicular trafficking. Modulation of receptor surface density during pathological processes is nowadays considered as an interesting therapeutic approach for management of diseases, such as chronic pain, in which an increased trafficking is associated with the pathological state. This review will focus on the recent advances trafficking of the thermoTRP channels, TRPV1, TRPV2, TRPV4, TRPM3, TRPM8 and TRPA1, into/from the plasma membrane. Particularly, regulated membrane insertion of thermoTRPs channels contributes to a fine tuning of final channel activity, and indeed, it has resulted in the development of novel therapeutic approaches with successful clinical results such as disruption of SNARE-dependent exocytosis by botulinum toxin or botulinomimetic peptides.

  16. 75 FR 64974 - Notice of Data Availability on Coal Combustion Residual Surface Impoundments

    Science.gov (United States)

    2010-10-21

    ... Coal Combustion Residual Surface Impoundments AGENCY: Environmental Protection Agency (EPA). ACTION... of Coal Combustion Residuals from Electric Utilities. The Agency is seeking public comment on how, if...; Disposal of Coal Combustion Residuals From Electric Utilities Docket, Attention Docket ID No.,...

  17. Denitrification 'hot spots' in soil following surface residue application

    Science.gov (United States)

    Kuntz, Marianne; Morley, Nicholas J.; Hallett, Paul D.; Watson, Christine; Baggs, Elizabeth M.

    2015-04-01

    The availability of organic C is an important driver for the production and reduction of the greenhouse gas nitrous oxide (N2O) during denitrification. Denitrification as a response to plant residue amendments to soil surfaces has been extensively researched. However, the nature of hotspot sites of N2O production and reduction within the soil profile, especially in relation to the location of applied residues, is unknown. In a laboratory experiment we investigated the relationship between denitrifier N2O surface fluxes and N2O production and reduction sites. Probes which equilibrate with the soil gas phase by diffusion were developed to quantify denitrification products and product ratios at 1-2 cm, 4.5-5.5 cm or 8-9 cm from the surface. 13C labelled barley straw was incorporated at rates of 0, 2 and 4 t ha-1 into the top 3 cm of soil and subsequently amended with 14NH415NO3. In a three week experiment the soil gas phase at the three depths was analysed for 15N-N2O, 15N-N2, 13C-CO2 and O2 concentrations. Additionally, cores were destructively sampled for mineral 15N as well as microbial C and dissolved C in the respective depths. 15N-N2O and CO2 surface fluxes peaked one day after N application, with residue application resulting in significantly higher 15N-N2O emission rates compared to the non-amended control. The timing of the 15N-N2O surface flux on day 1 was related to maximum 15N-N2O concentrations of 36.6 μg 15N L-1 within the pore space at 5 cm depth. Three days after fertilizer application 15N-N2O pore space concentrations had significantly increased to 193 μg 15N L-1 at 9 cm depth indicating denitrifier activity at greater depth. Denitrification below the soil surface could be explained by increased microbial activity, oxygen depletion with increasing depth and progressive downwards diffusion of fertilizer NO3-. However, C availability appeared to only affect denitrification in the surface layer in which the residue was incorporated. Our results provide

  18. Microscopic theory of the residual surface resistivity of Rashba electrons

    Science.gov (United States)

    Bouaziz, Juba; Lounis, Samir; Blügel, Stefan; Ishida, Hiroshi

    2016-07-01

    A microscopic expression of the residual electrical resistivity tensor is derived in linear response theory for Rashba electrons scattering at a magnetic impurity with cylindrical or noncylindrical potential. The behavior of the longitudinal and transversal residual resistivity is obtained analytically and computed for an Fe impurity at the Au(111) surface. We studied the evolution of the resistivity tensor elements as a function of the Rashba spin-orbit strength and the magnetization direction of the impurity. We found that the absolute values of longitudinal resistivity reduce with increasing spin-orbit strength of the substrate and that the scattering of the conduction electrons at magnetic impurities with magnetic moments pointing in directions not perpendicular to the surface plane produce a planar Hall effect and an anisotropic magnetoresistance even if the impurity carries no spin-orbit interaction. Functional forms are provided describing the anisotropy of the planar Hall effect and the anisotropic magnetoresistance with respect to the direction of the impurity moment. In the limit of no spin-orbit interaction and a nonmagnetic impurity of cylindrical symmetry, the expression of the residual resistivity of a two-dimensional electron gas has the same simplicity and form as for the three-dimensional electron gas [J. Friedel, J. Nuovo. Cim. 7, 287 (1958), 10.1007/BF02751483] and can also be expressed in terms of scattering phase shifts.

  19. Relation between the secondary structure of carbohydrate residues of alpha1-acid glycoprotein (orosomucoid) and the fluorescence of the protein.

    Science.gov (United States)

    Albani, Jihad R

    2003-05-01

    We studied in this work the relation that exists between the secondary structure of the glycans of alpha(1)-acid glycoprotein and the fluorescence of the Trp residues of the protein. We calculated for that the efficiency of quenching and the radiative and non-radiative constants. Our results indicate that the glycans display a spatial structure that is modified upon asialylation. The asialylated conformation is closer to the protein matrix than the sialylated form, inducing by that a decrease in the fluorescence parameters of the Trp residues. In fact, the mean quantum yield of Trp residues in sialylated and asialylated alpha(1)-acid glycoprotein are 0.0645 and 0.0385, respectively. Analysis of the fluorescence emission of alpha(1)-acid glycoprotein as the result of two contributions (surface and hydrophobic domains) indicates that quantum yields of both classes of Trp residues are lower when the protein is in the asialylated form. Also, the mean fluorescence lifetime of Trp residues decreases from 2.285 ns in the sialylated protein to 1.948 ns in the asialylated one. The radiative rate constant k(r) of the Trp residues in the sialylated alpha(1)-acid glycoprotein is higher than that in the asialylated protein. Thus, the carbohydrate residues are closer to the Trp residues in the absence of sialic acid. The modification of the spatial conformation of the glycans upon asialylation is confirmed by the decrease of the fluorescence lifetimes of Calcofluor, a fluorophore that binds to the carbohydrate residues. Finally, thermal intensity quenching of Calcofluor bound to alpha(1)-acid glycoprotein shows that the carbohydrate residues have slower residual motions in the absence of sialic acid residues.

  20. Analysis of the sexual development-promoting region of Schizophyllum commune TRP1 gene.

    Science.gov (United States)

    Sen, Kikuo; Kinoshita, Hideki; Tazuke, Kazuyuki; Maki, Yoshinori; Yoshiura, Yumi; Yakushi, Toshiharu; Shibai, Hiroshiro; Kurosawa, Shin-Ichi

    2016-10-01

    This study aims to elucidate the mechanism of sexual development of basidiomycetous mushrooms from mating to fruit body formation. Sequencing analysis showed the TRP1 gene of basidiomycete Schizophyllum commune encoded an enzyme with three catalytic regions of GAT (glutamine amidotransferase), IGPS (indole-3-glycerol phosphate synthase), and PRAI (5-phosphoribosyl anthranilate isomerase); among these three regions, the trp1 mutant (Trp(-)) had a missense mutation (L→F) of a 338th amino acid residue of the TRP1 protein within the IGPS region. To investigate the function of IGPS region related to sexual development, dikaryons with high, usual, and no expression of the IGPS region of TRP1 gene were made. The dikaryotic mycelia with high expression of the IGPS formed mature fruit bodies earlier than those with usual and no expression of the IGPS. These results showed that the IGPS region in TRP1 gene promoted sexual development of S. commune.

  1. Evaluation of the residue from microset on various metal surfaces.

    Energy Technology Data Exchange (ETDEWEB)

    Brumbach, Michael Todd

    2011-04-01

    Fast-curing impression materials are sometimes used to cast negative-mold replications of physical defects on material surfaces. The negative-mold impressions can then be used for further measurements to record the nature of the defect. These impression materials have been designed to cure quickly, and with very low adhesion, so that they can be easily removed from the surface leaving little residual contamination. Unfortunately, some contaminant is retained by the substrate material. This investigation seeks to identify the composition and quantity of the remaining material upon removal of Microset Synthetic Rubber Replicating Compound from several material surfaces. Coe-Flex was used as a relative comparison to Microset. On fifteen different substrate materials the Microset leaves no visible trace of contaminant, however, X-ray photoelectron spectroscopy shows evidence of a thin silicone-based contaminant film of approximately 2 nm thickness.

  2. Tryptophan 19 residue is the origin of bovine β-lactoglobulin fluorescence.

    Science.gov (United States)

    Albani, Jihad René; Vogelaer, Julie; Bretesche, Loïc; Kmiecik, Daniel

    2014-03-01

    β-Lactoglobulin consists of a single polypeptide of 162 amino acid residues with 2 Trp residues, Trp 19 present in a hydrophobic pocket and Trp 61 present at the surface of the protein near the pocket. This study aimed to characterize the respective contribution of the two Trp residues to the overall fluorescence of the protein. We added for that calcofluor white, an extrinsic fluorophore, which, at high concentration compared to that of the protein, quenches completely emission of hydrophobic Trp residue(s). The study was performed at different pHs by recording fluorescence steady state spectra and measuring fluorescence lifetimes of the Trp-residues using Single Time Photon Counting method. Our results indicate that addition of calcofluor white does not induce a red shift of the tryptophan(s) emission peak (332nm) but only a decrease in the fluorescence intensity. This means that Trp 61 residue does not contribute to the protein emission, tryptophan emission occurs from Trp 19 residue only. Also, excitation spectrum peak position (283nm) of β-lactoglobulin is not modified upon calcofluor white binding. These results mean that structural rearrangements within β-lactoglobulin are not occurring upon calcofluor white binding. Energy transfer between Trp 19 residue and calcofluor white occurs with 100% efficiency, i.e. the two fluorophores are very close one to each other (<5Å). This energy transfer is not Forster type. Fluorescence intensity decay of Trp 19 residue occurs with three lifetimes, equal to 0.48, 1.49 and 4.29ns at pH 2 (monomeric state). Very close values were obtained at the different studied pHs (2-12) and where β-lactoglobulin is at different quaternary structure or present in solution in a mixture of dimers and monomers. Our data are interpreted as the results of emission occurring from different substructures of the tryptophan, reached at the excited state. The populations of these substructures characterized by the pre-exponential parameters

  3. TRP channels and psychiatric disorders.

    Science.gov (United States)

    Chahl, Loris A

    2011-01-01

    Depression and schizophrenia are major psychiatric disorders that cause much human suffering. Current treatments have major limitations and new drug targets are eagerly sought. Study of transient receptor potential (TRP) channels in these disorders is at an early stage and the potential of agents that activate or inhibit these channels remains speculative. The findings that TRPC6 channels promote dendritic growth and are selectively activated by hyperforin, the key constitutent of St John's wort, suggest that TRPC6 channels might prove to be a new target for antidepressant drug development. There is now considerable evidence that TRPV1 antagonists have anxiolytic activity but there is no direct evidence that they have antidepressant activity. There is also no direct evidence that TRP channels play a role in schizophrenia. However, the findings that TRPC channels are involved in neuronal development and fundamental synaptic mechanisms, and that TRPV1 channels play a role in central dopaminergic and cannabinoid mechanisms is suggestive of potential roles of these channels in schizophrenia. Investigation of TRP channels in psychiatric disorders holds the promise of yielding further understanding of the aetiology of psychiatric disorders and the development of new drug treatments.

  4. Insertion mode of a novel anionic antimicrobial peptide MDpep5 (Val-Glu-Ser-Trp-Val) from Chinese traditional edible larvae of housefly and its effect on surface potential of bacterial membrane.

    Science.gov (United States)

    Tang, Ya-Li; Shi, Yong-Hui; Zhao, Wei; Hao, Gang; Le, Guo-Wei

    2008-12-01

    Antimicrobial molecules from insects may serve as a potentially significant group of antibiotics. To identify the effect of antimicrobial peptides (AMPs) on bacterial membrane and obtain further insight in the mechanism of membrane transport of AMPs, the interaction of surface potential and permeation of a novel antimicrobial peptide MDpep5 (Val-Glu-Ser-Trp-Val) from Chinese traditional edible larvae of housefly was examined using liposomes from bacterial lipids extract. Compared with the cationic AMPs, MDpep5 cannot completely disrupt membrane. The uptake of MDpep5 by bacterial liposomes was dependent on the membrane surface potential. The mutual inhibition of the transport of MDpep5 through the cell membrane was caused by the change in surface potential due to the binding of MDpep5 to the membrane. Furthermore, formation of MDpep5-enriched lipid aggregates could lead to the disorder of the bilayer structure. Based on our experimental data, we propose that MDpep5 initiated its antimicrobial activity by profoundly disordering the structure and affecting physical properties of bacterial membrane when binding to the phospholipid which accounts for its bactericidal activity.

  5. Finite Element Surface Layer Inheritable Condition Residual Stresses Model in Surface Plastic Deformation Processes

    Science.gov (United States)

    Mahalov, M. S.; Blumenstein, V. Yu

    2016-04-01

    The residual stresses (RS) research and computational algorithms creation in complex types of loading on the product lifecycle stages relevance is shown. The RS forming finite element model at surface plastic deformation strengthening machining, including technological inheritance effect, is presented. A model feature is the production previous stages obtained transformation properties consideration, as well as these properties evolution during metal particles displacement through the deformation space in the present loading step.

  6. Influence of hydrophobic and superhydrophobic surfaces on reducing aerodynamic insect residues

    Science.gov (United States)

    Krishnan, K. Ghokulla; Milionis, Athanasios; Loth, Eric; Farrell, Thomas E.; Crouch, Jeffrey D.; Berry, Douglas H.

    2017-01-01

    Insect fouling during takeoff, climb and landing can result in increased drag and fuel consumption for aircrafts with laminar-flow surfaces. This study investigates the effectiveness of various hydrophobic and superhydrophobic surfaces in reducing residue of insects on an aerodynamic surface at relatively high impact speeds (about 45 m/s). An experimental setup consisting of a wind tunnel and a method to inject live flightless fruit flies was used to test the effectiveness of various surfaces against insect fouling. Insect fouling was analyzed based on residue area and height from multiple impacts. In general most of the residue area was due to the hemolymph spreading while most of the residue height was due to adhesion of exoskeleton parts. Hydrophobic and especially superhydrophobic surfaces performed better than a hydrophilic aluminum surface in terms of minimizing the residue area of various insect components (exoskeleton, hemolymph, and red fluid). Surfaces with reduced wettability and short lateral length scales tended to have the smallest residue area. Residue height was not as strongly influenced by surface wettability since even a single exoskeleton adhered to the surface upon impact was enough to produce a residue height of the order of one mm. In general, the results indicate that hemolymph spread needs to be avoided (e.g. by having reduced wettability and short lateral correlation lengths) in order to minimize the residue area, while exoskeleton adherence needs to be avoided (e.g. by having oleophobic properties and micro/nano roughness) in order to minimize the residue height. In particular, two of the superhydrophobic coatings produced substantial reduction in residue height and area, relative to the baseline surface of aluminum. However, the surfaces also showed poor mechanical durability on the high-speed insect impact location. This suggests that although low wettability materials show great insect anti-fouling behavior, their durability needs to

  7. TRP Channels in Skin Biology and Pathophysiology

    OpenAIRE

    2016-01-01

    Ion channels of the Transient Receptor Potential (TRP) family mediate the influx of monovalent and/or divalent cations into cells in response to a host of chemical or physical stimuli. In the skin, TRP channels are expressed in many cell types, including keratinocytes, sensory neurons, melanocytes, and immune/inflammatory cells. Within these diverse cell types, TRP channels participate in physiological processes ranging from sensation to skin homeostasis. In addition, there is a growing body ...

  8. Finite Element Analysis and Experiment Research on Surface Residual Stress of Ceramics Grinding

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The theoretical model of residual stress of ceramics grinding has been established applying thermal elastoplastic mechanics theory. While grinding at the course of grinding wheel moved along workpiece surface the distributing regulation of residual stress can be simplified into thermal elastioplastic mechanical issue, under the action of the both moving centralized force and heat source. Calculating and evaluating of surface residual stress using current procedure of finite element analysis which has been...

  9. Chemical Modification of Tryptophan Residues in Superoxide Dismutase from Camellia Pollen and Its Fluorescence Spectrum

    Institute of Scientific and Technical Information of China (English)

    HE Xiao-hong; WU Min; LI Shan-yu; CHU Yu-zhuo; CHEN Jia; LIU Lan-ying

    2005-01-01

    The amino acid composition of the superoxide dismutase(SOD) from camellia pollen was measured and the tryptophan(Trp) residues were modified by using N-bromosuccinimide(NBS). The results show that there are 21 Trp residues in an SOD molecule and seven of which are located on the surface of the enzyme. By researching the fluorescence spectra of the native SOD and the modified SOD, we have found that the emission wavelength of Trp is at 335 nm and the fluorescence intensity will decrease when the enzyme is modified. The results also show that potassium iodide(KI) can significantly quench the fluorescence of the native SOD, but it has a less pronounced effect on the modified enzyme. Glycerin as a surface activation reagent can stabilize the fluorescence of the modified enzyme.

  10. Influence of Surface Properties and Impact Conditions on Adhesion of Insect Residues

    Science.gov (United States)

    Wohl, Christopher J.; Smith, Joseph G.; Connell, John W.; Siochi, Emilie J.; Doss, Jereme R.; Shanahan, Michelle H.; Penner, Ronald K.

    2015-01-01

    Insect residues can cause premature transition to turbulent flow on laminar flow airfoils. Engineered surfaces that mitigate the adhesion of insect residues provide, therefore, a route to more efficient aerodynamics and reduced fuel burn rates. Areal coverage and heights of residues depend not only on surface properties, but also on impact conditions. We report high speed photography of fruit fly impacts at different angles of inclination on a rigid aluminum surface, optical microscopy and profilometry, and contact angle goniometry to support the design of engineered surfaces. For the polyurethane and epoxy coatings studied, some of which exhibited superhydrophobicity, it was determined that impact angle and surface compositions play critical roles in the efficacy of these surfaces to reduce insect residue adhesion.

  11. TRP Channels in Skin Biology and Pathophysiology

    Directory of Open Access Journals (Sweden)

    Michael J. Caterina

    2016-12-01

    Full Text Available Ion channels of the Transient Receptor Potential (TRP family mediate the influx of monovalent and/or divalent cations into cells in response to a host of chemical or physical stimuli. In the skin, TRP channels are expressed in many cell types, including keratinocytes, sensory neurons, melanocytes, and immune/inflammatory cells. Within these diverse cell types, TRP channels participate in physiological processes ranging from sensation to skin homeostasis. In addition, there is a growing body of evidence implicating abnormal TRP channel function, as a product of excessive or deficient channel activity, in pathological skin conditions such as chronic pain and itch, dermatitis, vitiligo, alopecia, wound healing, skin carcinogenesis, and skin barrier compromise. These diverse functions, coupled with the fact that many TRP channels possess pharmacologically accessible sites, make this family of proteins appealing therapeutic targets for skin disorders.

  12. TRP Channels in Skin Biology and Pathophysiology

    Science.gov (United States)

    Caterina, Michael J.; Pang, Zixuan

    2016-01-01

    Ion channels of the Transient Receptor Potential (TRP) family mediate the influx of monovalent and/or divalent cations into cells in response to a host of chemical or physical stimuli. In the skin, TRP channels are expressed in many cell types, including keratinocytes, sensory neurons, melanocytes, and immune/inflammatory cells. Within these diverse cell types, TRP channels participate in physiological processes ranging from sensation to skin homeostasis. In addition, there is a growing body of evidence implicating abnormal TRP channel function, as a product of excessive or deficient channel activity, in pathological skin conditions such as chronic pain and itch, dermatitis, vitiligo, alopecia, wound healing, skin carcinogenesis, and skin barrier compromise. These diverse functions, coupled with the fact that many TRP channels possess pharmacologically accessible sites, make this family of proteins appealing therapeutic targets for skin disorders. PMID:27983625

  13. Experimental Research on Residual Stress in Surface of Silicon Nitride Ceramic Balls

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The influence of the residual stress in surface of ceramic balls on the fatigue life is large, because the life of silicon nitride ball bearings is more sensitive to the load acted on the bearings than the life of all-steel ball bearings. In this paper, the influence of thermal stress produced in sintering and mechanical stress formed in lapping process on residual stress in surface of silicon nitride ceramic balls was discussed. The residual compress stress will be formed in the surface of silicon nitride ...

  14. Surface analysis of carbon black waste materials from tire residues

    Science.gov (United States)

    Lee, W. H.; Kim, J. Y.; Ko, Y. K.; Reucroft, P. J.; Zondlo, J. W.

    1999-03-01

    X-ray photoelectron spectroscopy (XPS) has been used to obtain surface chemical state information on two carbon black waste materials in terms of the surface element distribution/concentration and chemical structure. Small amounts of sulfur in the form of CS 2 were detected on the surface (less than 1.7 mass %). C-H/C-C was the major carbon functional component on the surface of carbon black samples but other functional forms of carbon were also present such as CO and C-O. The surface of the carbon black obtained from a hydropyrolysis process was highly oxidized primarily in the form of carbon based oxygen groups. On the other hand, surface oxygen atoms on the surface of the carbon black obtained from a pyrolysis process in the absence of H 2 were in the form of both metal oxides and carbon based oxygen groups.

  15. Surface electrochemical properties of red mud (bauxite residue): zeta potential and surface charge density.

    Science.gov (United States)

    Liu, Yanju; Naidu, Ravendra; Ming, Hui

    2013-03-15

    The surface electrochemical properties of red mud (bauxite residue) from different alumina refineries in Australia and China were studied by electrophoresis and measuring surface charge density obtained from acid/base potentiometric titrations. The electrophoretic properties were measured from zeta potentials obtained in the presence of 0.01 and 0.001 M KNO(3) over a wide pH range (3.5-10) by titration. The isoelectric point (IEP) values were found to vary from 6.35 to 8.70 for the red mud samples. Further investigation into the surface charge density of one sample (RRM) by acid/base potentiometric titration showed similar results for pH(PZC) with pH(IEP) obtained from electrokinetic measurements. The pH(IEP) determined from zeta potential measurements can be used as a characteristic property of red mud. The minerals contained in red mud contributed to the different values of pH(IEP) of samples obtained from different refineries. Different relationships of pH(IEP) with Al/Fe and Al/Si ratios (molar basis) were also found for different red mud samples.

  16. Effect of the residual stresses on surface coercive force in amorphous alloys

    Energy Technology Data Exchange (ETDEWEB)

    Tejedor, M.; Garcia, J.A.; Carrizo, J.; Elbaile, L. E-mail: elbaile@pinon.ccu.uniovi.es; Santos, J.D

    2000-06-02

    The dependence of the coercivity H{sub c} on the applied stress {sigma} in the surfaces of various amorphous ribbons with positive magnetostriction is studied. The results obtained show that the behaviour of H{sub c}({sigma}) in the surfaces is the same as in the bulk and the minimum of H{sub c}({sigma}) does not depend on the residual stresses. The residual stresses affect the value of the coercive field but not its stress dependence.

  17. Experimental measurements of surface damage and residual stresses in micro-engineered plasma facing materials

    Science.gov (United States)

    Rivera, David; Wirz, Richard E.; Ghoniem, Nasr M.

    2017-04-01

    The thermomechanical damage and residual stresses in plasma-facing materials operating at high heat flux are experimentally investigated. Materials with micro-surfaces are found to be more resilient, when exposed to cyclic high heat flux generated by an arc-jet plasma. An experimental facility, dedicated to High Energy Flux Testing (HEFTY), is developed for testing cyclic heat flux in excess of 10 MW/m2. We show that plastic deformation and subsequent fracture of the surface can be controlled by sample cooling. We demonstrate that W surfaces with micro-pillar type surface architecture have significantly reduced residual thermal stresses after plasma exposure, as compared to those with flat surfaces. X-ray diffraction (XRD) spectra of the W-(110) peak reveal that broadening of the Full Width at Half Maximum (FWHM) for micro-engineered samples is substantially smaller than corresponding flat surfaces. Spectral shifts of XRD signals indicate that residual stresses due to plasma exposure of micro-engineered surfaces build up in the first few cycles of exposure. Subsequent cyclic plasma heat loading is shown to anneal out most of the built-up residual stresses in micro-engineered surfaces. These findings are consistent with relaxation of residual thermal stresses in surfaces with micro-engineered features. The initial residual stress state of highly polished flat W samples is compressive (≈ -1.3 GPa). After exposure to 50 plasma cycles, the surface stress relaxes to -1.0 GPa. Micro-engineered samples exposed to the same thermal cycling show that the initial residual stress state is compressive at (- 250 MPa), and remains largely unchanged after plasma exposure.

  18. Atomistic modelling of residual stress at UO2 surfaces.

    Science.gov (United States)

    Arayro, Jack; Tréglia, Guy; Ribeiro, Fabienne

    2016-01-13

    Modelling oxide surface behaviour is of both technological and fundamental interest. In particular, in the case of the UO2 system, which is of major importance in the nuclear industry, it is essential to account for the link between microstructure and macroscopic mechanical properties. Indeed micromechanical models at the mesoscale need to be supplied by the energetic and stress data calculated at the nanoscale. In this framework, we present a theoretical study, coupling an analytical model and thermostatistical simulation to investigate the modifications induced by the presence of a surface regarding atomic relaxation and energetic and stress profiles. In particular, we show that the surface effective thickness as well as the stress profile, which are required by micromechanical approaches, are strongly anisotropic.

  19. Catalytic domain surface residues mediating catecholamine inhibition in tyrosine hydroxylase.

    Science.gov (United States)

    Briggs, Gabrielle D; Bulley, Jesse; Dickson, Phillip W

    2014-03-01

    Tyrosine hydroxylase (TH) performs the rate-limiting step in catecholamine (CA) synthesis and is a tetramer composed of regulatory, catalytic and tetramerization domains. CAs inhibit TH by binding two sites in the active site; one with high affinity and one with low affinity. Only high affinity CA binding requires the regulatory domain, believed to interact with the catalytic domain in the presence of CA. Without a crystal structure of the regulatory domain, the specific areas involved in this process are largely undefined. It is not clear whether the regulatory domain-catalytic domain interaction is asymmetrical across the tetramer to produce the high and low affinity sites. To investigate this, pure dimeric TH was generated through double substitution of residues at the tetramerization interface and dimerization salt bridge (K170E/L480A). This was shown to be the core regulatory unit of TH for CA inhibition, possessing both high and low affinity CA binding sites, indicating that there is symmetry between dimers of the tetramer. We also examined possible regulatory domain-interacting regions on the catalytic domain that mediate high affinity CA binding. Using site-directed mutagenesis, A297, E362/E365 and S368 were shown to mediate high affinity dopamine inhibition through V(max) reduction and increasing the K(M) for the cofactor.

  20. Introduction of Enhanced Compressive Residual Stress Profiles in Aerospace Components Using Combined Mechanical Surface Treatments

    Science.gov (United States)

    Gopinath, Abhay; Lim, Andre; Nagarajan, Balasubramanian; Cher Wong, Chow; Maiti, Rajarshi; Castagne, Sylvie

    2016-11-01

    Mechanical surface treatments such as Shot Peening (SP) and Deep Cold Rolling (DCR) are being used to introduce Compressive Residual Stress (CRS) at the surface and subsurface layers of aerospace components, respectively. This paper investigates the feasibility of a combined introduction of both the surface and sub-surface compressive residual stress on Ti6Al4V material through a successive application of the two aforementioned processes, one after the other. CRS profiles between individual processes were compared to that of combination of processes to validate the feasibility. It was found out that shot peening introduces surface compressive residual stress into the already deep cold rolled sample, resulting in both surface and sub-surface compressive residual stresses in the material. However the drawback of such a combination would be the increased surface roughness after shot peening a deep cold rolled sample which can be critical especially in compressor components. Hence, a new technology, Vibro-Peening (VP) may be used as an alternative to SP to introduce surface stress at reduced roughness.

  1. Surface Finish and Residual Stresses Induced by Orthogonal Dry Machining of AA7075-T651.

    Science.gov (United States)

    Jomaa, Walid; Songmene, Victor; Bocher, Philippe

    2014-02-28

    The surface finish was extensively studied in usual machining processes (turning, milling, and drilling). For these processes, the surface finish is strongly influenced by the cutting feed and the tool nose radius. However, a basic understanding of tool/surface finish interaction and residual stress generation has been lacking. This paper aims to investigate the surface finish and residual stresses under the orthogonal cutting since it can provide this information by avoiding the effect of the tool nose radius. The orthogonal machining of AA7075-T651 alloy through a series of cutting experiments was performed under dry conditions. Surface finish was studied using height and amplitude distribution roughness parameters. SEM and EDS were used to analyze surface damage and built-up edge (BUE) formation. An analysis of the surface topography showed that the surface roughness was sensitive to changes in cutting parameters. It was found that the formation of BUE and the interaction between the tool edge and the iron-rich intermetallic particles play a determinant role in controlling the surface finish during dry orthogonal machining of the AA7075-T651 alloy. Hoop stress was predominantly compressive on the surface and tended to be tensile with increased cutting speed. The reverse occurred for the surface axial stress. The smaller the cutting feed, the greater is the effect of cutting speed on both axial and hoop stresses. By controlling the cutting speed and feed, it is possible to generate a benchmark residual stress state and good surface finish using dry machining.

  2. [Residue characteristics and distributions of perfluorinated compounds in surface seawater along Shenzhen coastline].

    Science.gov (United States)

    Chen, Qing-Wu; Zhang, Hong; Chai, Zhi-Fang; Shen, Jin-Can; Yang, Bo

    2012-06-01

    In order to explore the residue characteristics and distributions of 15 perfluorinated compounds (PFCs) in 18 surface seawater samples along Shenzhen coastline, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) combined with solid phase extraction enrichment was applied in this research. The results indicated that residue level of PFCs in coastal surface seawater samples was significantly affected by human activities. Sigma PFCs residue levels in surface seawater from Shenzhen west coast, which locates below the estuary of Pearl River and Donghao River, are much higher than those from the east coast, which has low development and sparse population (P<0.05). Under natural conditions, sigma PFCs residue levels in coastal surface seawater samples from Shenzhen Bays are higher than those out of bays. The major residue species in surface seawater samples along Shenzhen coast were medium- and short-chain PFCs, including perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorohexanoic acid and perfluoropentanoic acid. Their similar environmental behavior (P<0.05, P<0.01) is likely associated with the production process of PFCs-related products. Furthermore, cluster analysis results show that PFOS (R2 = 0.4092) level can be used as a representative parameter for evaluating PFCs contamination status in surface seawater along Shenzhen coast.

  3. Detection of organic residues on food processing equipment surfaces by spectral imaging method

    Science.gov (United States)

    Qin, Jianwei; Jun, Won; Kim, Moon S.; Chao, Kaunglin

    2010-04-01

    Organic residues on equipment surfaces in poultry processing plants can generate cross contamination and increase the risk of unsafe food for consumers. This research was aimed to investigate the potential of LED-induced fluorescence imaging technique for rapid inspection of organic residues on poultry processing equipment surfaces. High-power blue LEDs with a spectral output at 410 nm were used as the excitation source for a line-scanning hyperspectral imaging system. Common chicken residue samples including fat, blood, and feces from ceca, colon, duodenum, and small intestine were prepared on stainless steel sheets. Fluorescence emission images were acquired from 120 samples (20 for each type of residue) in the wavelength range of 500-700 nm. LED-induced fluorescence characteristics of the tested samples were determined. PCA (principal component analysis) was performed to analyze fluorescence spectral data. Two SIMCA (soft independent modeling of class analogy) models were developed to differentiate organic residues and stainless steel samples. Classification accuracies using 2-class ('stainless steel' and 'organic residue') and 4-class ('stainless steel', 'fat', 'blood', and 'feces') SIMCA models were 100% and 97.5%, respectively. An optimal single-band and a band-pair that are promising for rapid residue detection were identified by correlation analysis. The single-band approach using the selected wavelength of 666 nm could generate false negative errors for chicken blood inspection. Two-band ratio images using 503 and 666 nm (F503/F666) have great potential for detecting various chicken residues on stainless steel surfaces. This wavelength pair can be adopted for developing a LED-based hand-held fluorescence imaging device for inspecting poultry processing equipment surfaces.

  4. Near-surface residual stresses and microstructural changes after turning of a nickel-based superalloy

    Energy Technology Data Exchange (ETDEWEB)

    Schlauer, Christian

    2003-07-01

    Nickel-based superalloys are precipitation hardened alloys with complex compositions. They are used in aircraft engines and land-based gas turbines in load bearing structural components that are exposed to high temperatures. Failure mechanisms in this environment are high and low cycle fatigue, creep, and corrosion. During manufacturing, residual stresses are often introduced into the material due to inhomogeneous plastic deformations, both intentionally and unintentionally. One such manufacturing process is metal cutting, which introduces residual stresses in the surface layer. The stress state in the near-surface zone of components is of special interest as the surface often experiences peak loads and cracks have their starting point there. In this thesis, near-surface residual stress distributions and microstructural changes are studied in the nickel-based superalloy Inconel 718 for two different turning operations, face grooving and facing. Process variables are in both cases cutting speed and feed that have been varied between (10 and 1200) m/min and (0.01 and 0.5) mm, respectively. The first turning technique face grooving, which gives cutting conditions similar to orthogonal cutting, showed a clear dependency of the residual stresses on the cutting speed. The tensile stress at the surface, the maximum compressive stress below the surface, and the thickness of the affected layer increase with increasing cutting speed. The tensile stresses are constrained to a thin surface layer and compressive residual stresses below the surface dominate the depth profile of the residual stresses. Only at low cutting speed, residual stresses were largely avoided. The second turning technique facing confirmed the dependency of the residual stresses on the cutting speed and revealed a similar dependency on the feed. Microstructural investigations of near-surface cross-sections by means of transmission electron microscopy showed a zone where the grains had undergone plastic

  5. Targeting TRP channels for novel migraine therapeutics.

    Science.gov (United States)

    Dussor, Gregory; Yan, J; Xie, Jennifer Y; Ossipov, Michael H; Dodick, David W; Porreca, Frank

    2014-11-19

    Migraine is increasingly understood to be a disorder of the brain. In susceptible individuals, a variety of "triggers" may influence altered central excitability, resulting in the activation and sensitization of trigeminal nociceptive afferents surrounding blood vessels (i.e., the trigeminovascular system), leading to migraine pain. Transient receptor potential (TRP) channels are expressed in a subset of dural afferents, including those containing calcitonin gene related peptide (CGRP). Activation of TRP channels promotes excitation of nociceptive afferent fibers and potentially lead to pain. In addition to pain, allodynia to mechanical and cold stimuli can result from sensitization of both peripheral afferents and of central pain pathways. TRP channels respond to a variety of endogenous conditions including chemical mediators and low pH. These channels can be activated by exogenous stimuli including a wide range of chemical and environmental irritants, some of which have been demonstrated to trigger migraine in humans. Activation of TRP channels can elicit CGRP release, and blocking the effects of CGRP through receptor antagonism or antibody strategies has been demonstrated to be effective in the treatment of migraine. Identification of approaches that can prevent activation of TRP channels provides an additional novel strategy for discovery of migraine therapeutics.

  6. Human salivary alpha-amylase Trp58 situated at subsite -2 is critical for enzyme activity.

    Science.gov (United States)

    Ramasubbu, Narayanan; Ragunath, Chandran; Mishra, Prasunkumar J; Thomas, Leonard M; Gyémánt, Gyöngyi; Kandra, Lili

    2004-06-01

    The nonreducing end of the substrate-binding site of human salivary alpha-amylase contains two residues Trp58 and Trp59, which belong to beta2-alpha2 loop of the catalytic (beta/alpha)(8) barrel. While Trp59 stacks onto the substrate, the exact role of Trp58 is unknown. To investigate its role in enzyme activity the residue Trp58 was mutated to Ala, Leu or Tyr. Kinetic analysis of the wild-type and mutant enzymes was carried out with starch and oligosaccharides as substrates. All three mutants exhibited a reduction in specific activity (150-180-fold lower than the wild type) with starch as substrate. With oligosaccharides as substrates, a reduction in k(cat), an increase in K(m) and distinct differences in the cleavage pattern were observed for the mutants W58A and W58L compared with the wild type. Glucose was the smallest product generated by these two mutants in the hydrolysis oligosaccharides; in contrast, wild-type enzyme generated maltose as the smallest product. The production of glucose by W58L was confirmed from both reducing and nonreducing ends of CNP-labeled oligosaccharide substrates. The mutant W58L exhibited lower binding affinity at subsites -2, -3 and +2 and showed an increase in transglycosylation activity compared with the wild type. The lowered affinity at subsites -2 and -3 due to the mutation was also inferred from the electron density at these subsites in the structure of W58A in complex with acarbose-derived pseudooligosaccharide. Collectively, these results suggest that the residue Trp58 plays a critical role in substrate binding and hydrolytic activity of human salivary alpha-amylase.

  7. Effect of shot peening and grit blasting on surface integrity: Influence on residual stresses

    Institute of Scientific and Technical Information of China (English)

    K.TOSHA; LU Jian

    2006-01-01

    The influences of factors such as particle size (0.55-2.2 mm), particle velocity (15-35 m/s) and thickness of work material on the surface integrity were investigated. The residual stresses induced by shot peening or grit blasting were examined. In order to clarify the influences of those factors on residual stress included in the surface integrity, a medium carbon steel (w(C)= 0.45%, 180 HV) was peened by a centrifugal type peening machine using cast steel particles (650-800 HV). The results show that the compressive residual stresses on the peened surface are larger than those of grit blasting; the critical thickness of shot peening is about 50% thicker than that of grit blasting; the high compressive stresses induced by blasting are owing to the wrought or peening effect.

  8. Insect Residue Contamination on Wing Leading Edge Surfaces: A Materials Investigation for Mitigation

    Science.gov (United States)

    Lorenzi, Tyler M.; Wohl, Christopher J.; Penner, Ronald K.; Smith, Joseph G.; Siochi, Emilie J.

    2011-01-01

    Flight tests have shown that residue from insect strikes on aircraft wing leading edge surfaces may induce localized transition of laminar to turbulent flow. The highest density of insect populations have been observed between ground level and 153 m during light winds (2.6 -- 5.1 m/s), high humidity, and temperatures from 21 -- 29 C. At a critical residue height, dependent on the airfoil and Reynolds number, boundary layer transition from laminar to turbulent results in increased drag and fuel consumption. Although this represents a minimal increase in fuel burn for conventional transport aircraft, future aircraft designs will rely on maintaining laminar flow across a larger portion of wing surfaces to reduce fuel burn during cruise. Thus, insect residue adhesion mitigation is most critical during takeoff and initial climb to maintain laminar flow in fuel-efficient aircraft configurations. Several exterior treatments investigated to mitigate insect residue buildup (e.g., paper, scrapers, surfactants, flexible surfaces) have shown potential; however, implementation has proven to be impractical. Current research is focused on evaluation of wing leading edge surface coatings that may reduce insect residue adhesion. Initial work under NASA's Environmentally Responsible Aviation Program focused on evaluation of several commercially available products (commercial off-the-shelf, COTS), polymers, and substituted alkoxy silanes that were applied to aluminum (Al) substrates. Surface energies of these coatings were determined from contact angle data and were correlated to residual insect excrescence on coated aluminum substrates using a custom-built "bug gun." Quantification of insect excrescence surface coverage was evaluated by a series of digital photographic image processing techniques.

  9. Residual strain evaluation of curved surface by grating-transferring technique and GPA

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    This paper investigates an advanced grating-transferring technique combined with geometric phase analysis (GPA) for residual strain evaluation of curved surface.A standard holographic grating is first transferred to a pre-produced epoxy resin film and then consolidated to a test region of curved surface.With a rubber mold and silicone rubber the deformed grating is replicated to a sheet metal after hole-drilling for release of residual stress.After that the grating is transferred from the sheet metal to the...

  10. Mechanosensitive TRP channels in cardiovascular pathophysiology.

    Science.gov (United States)

    Inoue, Ryuji; Jian, Zhong; Kawarabayashi, Yasuhiro

    2009-09-01

    Transient receptor potential (TRP) proteins constitute a large non-voltage-gated cation channel superfamily, activated polymodally by various physicochemical stimuli, and are implicated in a variety of cellular functions. Known activators for TRP include not only chemical stimuli such as receptor stimulation, increased acidity and pungent/cooling agents, but temperature change and various forms of mechanical stimuli such as osmotic stress, membrane stretch, and shear force. Recent investigations have revealed that at least ten mammalian TRPs exhibit mechanosensitivity (TRPC1, 5, 6; TRPV1, 2, 4; TRPM3, 7; TRPA1; TRPP2), but the mechanisms underlying it appear considerably divergent and complex. The proposed mechanisms are associated with lipid bilayer mechanics, specialized force-transducing structures, biochemical reactions, membrane trafficking and transcriptional regulation. Many of mechanosensitive (MS)-TRP channel likely undergo multiple regulations via these mechanisms. In the cardiovascular system in which hemodynamic forces constantly operate, the impact of mechanical stress may be particularly significant. Extensive morphological and functional studies have indicated that several MS-TRP channels are expressed in cardiac muscle, vascular smooth muscle, endothelium and vasosensory neurons, each differentially contributing to cardiovascular (CV) functions. To further complexity, the recent evidence suggests that mechanical stress may synergize with neurohormonal mechanisms thereby amplifying otherwise marginal responses. Furthermore, the currently available data suggest that MS-TRP channels may be involved in CV pathophysiology such as cardiac arrhythmia, cardiac hypertrophy/myopathy, hypertension and aneurysms. This review will overview currently known mechanisms for mechanical activation/modulation of TRPs and possible connections of MS-TRP channels to CV disorders.

  11. The role of basic residues in the adsorption of blood proteins onto the graphene surface

    Science.gov (United States)

    Gu, Zonglin; Yang, Zaixing; Wang, Lingle; Zhou, Hong; Jimenez-Cruz, Camilo A.; Zhou, Ruhong

    2015-06-01

    With its many unique properties, graphene has shown great potential in various biomedical applications, while its biocompatibility has also attracted growing concerns. Previous studies have shown that the formation of protein-graphene corona could effectively reduce its cytotoxicity; however, the underlying molecular mechanism remains not well-understood. Herein, we use extensive molecular dynamics simulations to demonstrate that blood proteins such as bovine fibrinogen (BFG) can absorb onto the graphene surface quickly and tightly to form a corona complex. Aromatic residues contributed significantly during this adsorption process due to the strong π-π stacking interactions between their aromatic rings and the graphene sp2-carbons. Somewhat surprisingly, basic residues like arginine, also played an equally or even stronger role during this process. The strong dispersion interactions between the sidechains of these solvent-exposed basic residues and the graphene surface provide the driving force for a tight binding of these basic residues. To the best of our knowledge, this is the first study with blood proteins to show that, in addition to the aromatic residues, the basic residues also play an important role in the formation of protein-graphene corona complexes.

  12. Analysis list: Trp53 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Trp53 Embryo,Embryonic fibroblast + mm9 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/target/Trp53....1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/target/Trp53.5.tsv http://dbarchive.bi...osciencedbc.jp/kyushu-u/mm9/target/Trp53.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/colo/Trp53.Em...bryo.tsv,http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/colo/Trp53.Embryonic_fibr

  13. Surface Finish and Residual Stresses Induced by Orthogonal Dry Machining of AA7075-T651

    Directory of Open Access Journals (Sweden)

    Walid Jomaa

    2014-02-01

    Full Text Available The surface finish was extensively studied in usual machining processes (turning, milling, and drilling. For these processes, the surface finish is strongly influenced by the cutting feed and the tool nose radius. However, a basic understanding of tool/surface finish interaction and residual stress generation has been lacking. This paper aims to investigate the surface finish and residual stresses under the orthogonal cutting since it can provide this information by avoiding the effect of the tool nose radius. The orthogonal machining of AA7075-T651 alloy through a series of cutting experiments was performed under dry conditions. Surface finish was studied using height and amplitude distribution roughness parameters. SEM and EDS were used to analyze surface damage and built-up edge (BUE formation. An analysis of the surface topography showed that the surface roughness was sensitive to changes in cutting parameters. It was found that the formation of BUE and the interaction between the tool edge and the iron-rich intermetallic particles play a determinant role in controlling the surface finish during dry orthogonal machining of the AA7075-T651 alloy. Hoop stress was predominantly compressive on the surface and tended to be tensile with increased cutting speed. The reverse occurred for the surface axial stress. The smaller the cutting feed, the greater is the effect of cutting speed on both axial and hoop stresses. By controlling the cutting speed and feed, it is possible to generate a benchmark residual stress state and good surface finish using dry machining.

  14. A residual life prediction model based on the generalized σ -N curved surface

    Directory of Open Access Journals (Sweden)

    Zongwen AN

    2016-06-01

    Full Text Available In order to investigate change rule of the residual life of structure under random repeated load, firstly, starting from the statistic meaning of random repeated load, the joint probability density function of maximum stress and minimum stress is derived based on the characteristics of order statistic (maximum order statistic and minimum order statistic; then, based on the equation of generalized σ -N curved surface, considering the influence of load cycles number on fatigue life, a relationship among minimum stress, maximum stress and residual life, that is the σmin(n- σmax(n-Nr(n curved surface model, is established; finally, the validity of the proposed model is demonstrated by a practical case. The result shows that the proposed model can reflect the influence of maximum stress and minimum stress on residual life of structure under random repeated load, which can provide a theoretical basis for life prediction and reliability assessment of structure.

  15. Engineering the residual stress state and microstructure of stainless steel with mechanical surface treatments

    Energy Technology Data Exchange (ETDEWEB)

    Turski, M.; Clitheroe, S.; Withers, P.J. [Manchester University, School of Materials, Manchester (United Kingdom); Evans, A.D. [Paul Scherrer Institut, Villigen-PSI (Switzerland); Rodopoulos, C. [University of Patras, Patras (Greece); Hughes, D.J. [Institut Laue Langevin, Grenoble (France)

    2010-06-15

    Four mechanical surface treatments have been considered for the application to austenitic stainless steel structures. Shot peening (SP), laser shock peening (LSP), ultrasonic impact treatment (UIT) and water jet cavitation peening (WJCP), also known as cavitation shotless peening (CSP), have been applied to 8 mm thick Type 304 austenitic stainless steel coupons. This study considers the merits of each of these mechanical surface treatments in terms of their effect on the surface roughness, microstructure, level of plastic work and through thickness residual stress distribution. Microstructural studies have revealed the formation of martensite close to the treated surface for each process. Residual stress measurements in the samples show compressive stresses to a significantly greater depth for the LSP, UIT and WJCP samples compared to the more conventional SP treated sample. (orig.)

  16. Side chain flexibility and coupling between the S4-S5 linker and the TRP domain in thermo-sensitive TRP channels: Insights from protein modeling.

    Science.gov (United States)

    Romero-Romero, Sergio; Gomez Lagunas, Froylan; Balleza, Daniel

    2017-04-01

    The transient receptor potential (TRP) superfamily is subdivided into several subfamilies on the basis of sequence similarity, which is highly heterogeneous but shows a molecular architecture that resembles the one present in members of the Kv channel superfamily. Because of this diversity, they produce a large variety of channels with different gating and permeability properties. Elucidation of these particular features necessarily requires comparative studies based on structural and functional data. The present study aims to compilate, analyze, and determine, in a coherent way, the relationship between intrinsic side-chain flexibility and the allosteric coupling in members of the TRPV, TRPM, and TRPC families. Based on the recently determined structures of TRPV1 and TRPV2, we have generated protein models for single subunits of TRPV5, TRPM8, and TRPC5 channels. With these models, we focused our attention on the apparently crucial role of the GP dipeptide at the center of the S4-S5 linker and discussed its role in the interaction with the TRP domain, specifically with the highly-conserved Trp during this coupling. Our analysis suggests an important role of the S4-S5L flexibility in the thermosensitivity, where heat-activated channels possess rigid S4-S5 linkers, whereas cold-activated channels have flexible ones. Finally, we also present evidence of the key interaction between the conserved Trp residue of the TRP box and of several residues in the S4-S5L, importantly the central Pro. Proteins 2017; 85:630-646. © 2016 Wiley Periodicals, Inc.

  17. Residual stresses in the surface layer of laser-treated steels

    NARCIS (Netherlands)

    Brussel, B.A. van; Hosson, J.Th.M. De

    1993-01-01

    Although laser treatment of certain metals may enhance the wear performance in general it may result equally well in large residual stresses which affect the wear performance detrimentally. Tensile stresses generated in the surface layer may lead to severe cracking of the material. This paper

  18. Cropping sequence and nitrogen fertilization impact on surface residue, soil carbon sequestration, and crop yields

    Science.gov (United States)

    Information is needed on the effect of management practices on soil C storage for obtaining C credit. The effects of tillage, cropping sequence, and N fertilization were evaluated on dryland crop and surface residue C and soil organic C (SOC) at the 0-120 cm depth in a Williams loam from 2006 to 201...

  19. Novel Epoxy Particulate Composites for Mitigation of Insect Residue Adhesion on Future Aircraft Surfaces

    Science.gov (United States)

    Wohl, Christopher J.; Smith, Joseph G., Jr.; Gardner, John M.; Penner, Ronald K.; Connell, John W.; Siochi, Emilie J.

    2014-01-01

    Drag is reduced significantly for airflow over surfaces when laminar flow can be maintained over greater chord lengths, the distance from the leading edge of an airfoil.1 However, surface imperfections, such as chipped paint, scratches, and events that change topography on a microscopic scale can introduce airflow instabilities resulting in premature transition to turbulent flow.1 Although many of these surface imperfections can be avoided with proper maintenance, advanced materials, and advanced manufacturing practices, topographical surface anomalies arising during flight from insect impacts cannot be controlled and can influence laminar flow stability. Practical solutions to this operational challenge need to be developed for future aircraft to have full advantage of laminar flow designs that improve fuel efficiency.2 Researchers have investigated various methods to mitigate insect residue adhesion for decades.3 Although several techniques have demonstrated efficacy including mechanical scrapers, active liquid discharge systems, and sacrificial paper coatings, they have not been commercially implemented due to increased manufacturing and operational complexity, environmental impact, and weight penalties. Coatings offer a simple route for passive insect residue adhesion prevention without many of the challenges associated with maintenance of laminar flow.4 In our previous work, we determined that most commercially available materials were not effective at insect residue adhesion.5 We also identified improvements when both surface energy could be controlled by surface modifying agents and the topography could be altered through the use of micron-sized and nanometer-sized filler materials.6 In this work, these general principles were applied to an epoxy system to evaluate the behavior of the surface modifying agent, a fluorinated alkyl ether oligomer, on surface energy and insect residue adhesion properties.

  20. Biochar production from coffee residues: Optimization of surface characteristics and sorptive behavior

    Science.gov (United States)

    Fotopoulou, Kalliopi; Manariotis, Ioannis D.; Karapanagioti, Hrissi K.

    2015-04-01

    Biochar with high surface area is a promising sorbent for environmental remediation and is produced by heating biomass in an oxygen-limited environment. Knowing the surface characteristics increases our understanding of biochar interactions with pollutants. The hypothesis of the present study is that by controlling pyrolysis conditions, the surface characteristics and subsequently the sorption behavior of produced biochars can be optimized. Coffee residues were dried overnight at 50oC and then pyrolized into a gradient furnace at 850oC. Different solid/oxygen ratios during pyrolysis were tested as well as the up scaling of the process. The biochars produced were systematically characterized for their surface characteristics such as BET surface area, open surface area, pore and micropore volume, and average pore size. The effect of pyrolysis on the biochar suspension pH was examined with the mass addition technique that involves the addition of increasing amounts of the biochar to bottles containing 0.1 M NaNO3. FTIR analysis was used in order to determine the functional groups of the coffee residue and of the biochars. The macrostructure of the biochars was visualized by Scanning Electron Microscopy (SEM). Total Carbon (TC) in the samples was determined by Carlo Erba Elemental Analyzer CHNS, EO 1108 after calibration with standard samples. The sorption behavior of produced biochars was tested with two different pollutants (Hg(II), phenanthrene) using batch reactors with the same initial single-compound solution and the same mass of coffee residue and different biochars. The biochars produced exhibited a wide range of surface area from 21 to 770 m2/g and open surface area due to macropores from 21 to 65 m2/g. This suggests that the surface area in the biochars with high surface area results from the formation of pores. Actually for the biochar with the highest surface area, it was calculated that up to 90

  1. An Amino Acid Coordination Polymer[Co(L-trp)(D-trp)]n%氨基酸配位聚合物[Co(L-trp)(D-trp)]n

    Institute of Scientific and Technical Information of China (English)

    王建; 许兴友; 马卫兴; 胡喜兰; 施鹏飞; 王明艳; 陆路德; 杨旭杰; 汪信

    2008-01-01

    The coordination polymer [Co(L-trp)(D-trp)]n (where L-trp is L-tryptophan or (S)-2-Amino-3-(3-indolyl)propionic acid and D-trp is D-tryptophan or (R)-2-Amino-3-(3-indolyl)propionic acid) was prepared by hydrothermal method. The structure and physicochemical properties were characterized by single-crystal X-ray diffraction structure determination, infrared spectroscopy, elemental analysis and cyclic vohammogram. The crystal data for the title compound: Monoclinic, P21/c, β=98.024(2)°, α=1.9595(3) nm, b=0.546 45(7) nm, c=0.905 54(17) nm, Z=2,μ=0.934 mm-1, R1=0.063 2, wR2=0. 155 3. The crystal structure shows a two-dimensional double chain plane structure feature. The cyrstallized coordination polymer has a centrosymmetric space group. Each Co2+ is coordinated with a pair of racemes, and the complex is a mesomer. CCDC: 651088.

  2. [Quantitative Analysis of Dimethoate Pesticide Residues in Honey by Surface-Enhanced Raman Spectroscopy].

    Science.gov (United States)

    Sun, Xu-dong; Dong, Xiao-ling

    2015-06-01

    The feasibility of a combination method of surface-enhanced Raman spectroscopy (SERS) technology and linear regression algorithm was investigated for rapid quantitative analysis of pesticide residues in honey. The total of 30 samples was applied in the experiment with dimethoate pesticide residues range from 1 ppm to 10 ppm. The samples were divided into calibration set (20) and prediction set (10). The substrate of Klarite with an inverted pyramidal structure was adopted for improvement of the relative intensity of the majority of Raman shift peaks. The comparative analysis was carried out between SERS spectra of dimethoate pesticide residues in honey samples and conventional Raman spectra of dimethoate standard sample. And four characteristic Raman shift peaks at the wavenumbers of 867, 1 065, 1 317 and 1 453 cm(-1) were found, which were related with the vibrational information of dimethoate molecule. The relationship was developed by linear regression algorithm between the intensity of Raman shift and the concentration of dimethoate pesticide residues. The 10 new samples in the prediction set were applied to evaluate the performance of the models. By comparison, the optimal model was obtained with the characteristic Raman shift peak of 867 cm(-1). The higher correlation coefficient of prediction of 0.984 and lower root mean square error of prediction of 0.663 ppm were obtained. The detection limit of this method was 2 ppm, which was close to the maximum levels of pesticide residue detection limits. Experimental results showed that it was feasible to rapidly analyze quantitative of pesticide residues in honey with the combination method of SERS technology and linear regression algorithm. Compared with the conventional method coupled with the suitable pretreatment, the combination method of SERS technology and linear regression method could analyze the dimethoate pesticide residues in honey, and it also provided an optional method for rapid quantitative analysis

  3. Antioxidant and antiglycation activities of the synthesised dipeptide, Asn-Trp, derived from computer-aided simulation of yam dioscorin hydrolysis and its analogue, Gln-Trp.

    Science.gov (United States)

    Han, Chuan-Hsiao; Lin, Yin-Shiou; Lin, Shyr-Yi; Hou, Wen-Chi

    2014-03-15

    Previous studies have shown that the Trp residue contributes to a high oxygen radical absorbance capacity (ORAC). Therefore, in this study, a Trp-containing dipeptide derived from a computer-aided simulation of pepsin hydrolysis of the yam tuber, dioscorin-namely, Asn-Trp (NW), and its analogue, Gln-Trp (QW)-were synthesised to compare their antioxidant and antiglycation activities with carnosine, homocarnosine, or glutathione (GSH). The antioxidant assays included hydroxyl radical-scavenging activity, anti-AAPH-induced hemolysis, and ORAC activity. NW had a significantly higher antioxidant activity than had QW and performed much better than carnosine, homocarnosine, or GSH. Using bovine serum albumin (BSA)/galactose or BSA/glucose as experimental models, NW had better antiglycation effects than had QW, as detected by an anti-N(ε)-(carboxymethyl)lysine (anti-CML) antibody. Moreover, NW and QW (50-200 μM) showed protection against methylglyoxal-induced cell deaths in human umbilical vein endothelial cells. These results suggest that NW, derived from computer-aided simulation of dioscorin hydrolysis, exhibits antioxidant and antiglycation activities, which thus shows the benefits of the yam tuber as an antioxidant-rich food. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Surface roughness due to residual ice in the use of low power deicing systems

    Science.gov (United States)

    Shin, Jaiwon; Bond, Thomas H.

    1993-01-01

    Thicknesses of residual ice are presented to provide information on surface contamination and associated roughness during deicing events. Data was obtained from low power ice protection systems tests conducted in the Icing Research Tunnel at NASA Lewis Research Center (LeRC) with nine different deicing systems. Results show that roughness associated with residual ice is not characterized by uniformly distributed roughness. Results also show that deicing systems require a critical mass of ice to generate a sufficient expelling force to remove the ice.

  5. SEM Analysis of Residual Dentin Surface in Primary Teeth Using Different Chemomechanical Caries Removal Agents.

    Science.gov (United States)

    Thakur, Rachna; Patil, Sandya Devi S; Kush, Anil; Madhu, K

    The purpose of this in vitro study was to analyze the residual dentinal surfaces following caries removal using two chemomechanical methods (Papacarie Duo and Carie Care), by scanning electron microscopy (SEM). Twenty extracted primary molars with active occlusal carious lesions were randomly assigned two groups depending on the CMCR agent used for the caries excavation - Group 1 - with Papacarie Duo and Group - 2 with Carie Care. After the caries excavation, the specimens were subjected to SEM analysis. Though both the agents showed the minimal smear layer with the patent dentinal tubules, Carie care showed patent dentinal tubules with a clearly exposed peritubular and intertubular collagen network. Carie Care treated surface exhibited better surface morphology of residual dentin.

  6. Influence of Initial Inclined Surface Crack on Estimated Residual Fatigue Lifetime of Railway Axle

    Science.gov (United States)

    Náhlík, Luboš; Pokorný, Pavel; Ševčík, Martin; Hutař, Pavel

    2016-11-01

    Railway axles are subjected to cyclic loading which can lead to fatigue failure. For safe operation of railway axles a damage tolerance approach taking into account a possible defect on railway axle surface is often required. The contribution deals with an estimation of residual fatigue lifetime of railway axle with initial inclined surface crack. 3D numerical model of inclined semi-elliptical surface crack in railway axle was developed and its curved propagation through the axle was simulated by finite element method. Presence of press-fitted wheel in the vicinity of initial crack was taken into account. A typical loading spectrum of railway axle was considered and residual fatigue lifetime was estimated by NASGRO approach. Material properties of typical axle steel EA4T were considered in numerical calculations and lifetime estimation.

  7. EFFECT OF SOIL TILLAGE AND PLANT RESIDUE ON SURFACE ROUGHNESS OF AN OXISOL UNDER SIMULATED RAIN

    Directory of Open Access Journals (Sweden)

    Elói Panachuki

    2015-02-01

    Full Text Available Surface roughness of the soil is formed by mechanical tillage and is also influenced by the kind and amount of plant residue, among other factors. Its persistence over time mainly depends on the fundamental characteristics of rain and soil type. However, few studies have been developed to evaluate these factors in Latossolos (Oxisols. In this study, we evaluated the effect of soil tillage and of amounts of plant residue on surface roughness of an Oxisol under simulated rain. Treatments consisted of the combination of the tillage systems of no-tillage (NT, conventional tillage (CT, and minimum tillage (MT with rates of plant residue of 0, 1, and 2 Mg ha-1 of oats (Avena strigosa Schreb and 0, 3, and 6 Mg ha-1 of maize (Zea mays L.. Seven simulated rains were applied on each experimental plot, with intensity of 60±2 mm h-1 and duration of 1 h at weekly intervals. The values of the random roughness index ranged from 2.94 to 17.71 mm in oats, and from 5.91 to 20.37 mm in maize, showing that CT and MT are effective in increasing soil surface roughness. It was seen that soil tillage operations carried out with the chisel plow and the leveling disk harrow are more effective in increasing soil roughness than those carried out with the heavy disk harrow and leveling disk harrow. The roughness index of the soil surface decreases exponentially with the increase in the rainfall volume applied under conditions of no tillage without soil cover, conventional tillage, and minimum tillage. The oat and maize crop residue present on the soil surface is effective in maintaining the roughness of the soil surface under no-tillage.

  8. Structure-activity relationship of Trp-containing analogs of the antimicrobial peptide gomesin.

    Science.gov (United States)

    Domingues, Tatiana M; Buri, Marcus V; Daffre, Sirlei; Campana, Patricia T; Riske, Karin A; Miranda, Antonio

    2014-06-01

    Gomesin (Gm) has a broad antimicrobial activity making it of great interest for development of drugs. In this study, we analyzed three Gm analogs, [Trp(1) ]-Gm, [Trp(7) ]-Gm, and [Trp(9) ]-Gm, in an attempt to gain insight into the contributions of different regions of the peptide sequence to its activity. The incorporation of the tryptophan residue in different positions has no effect on the antimicrobial and hemolytic activities of the Gm analogs in relation to Gm. Spectroscopic studies (circular dichroism, fluorescence and absorbance) of Gm and its analogs were performed in the presence of SDS, below and above its critical micelle concentration (CMC) (~8 mM), in order to monitor structural changes induced by the interaction with this anionic surfactant (0-15 mM). Interestingly, we found that the analogs interact more strongly with SDS at low concentrations (0.3-6.0 mM) than close to or above its CMC. This suggests that SDS monomers are able to cover the whole peptide, forming large detergent-peptide aggregates. On the other hand, the peptides interact differently with SDS micelles, inserting partially into the micelle core. Among the peptides, Trp in position 1 becomes more motionally-restricted in the presence of SDS, probably because this residue is located at the N-terminal region, which presents higher conformational freedom to interact stronger with SDS molecules. Trp residues in positions 7 and 9, close to and in the region of the turn of the molecule, respectively, induced a more constrained structure and the compounds cannot insert deeper into the micelle core or be completely buried by SDS monomers. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.

  9. Species-specific identity elements of tRNA Trp

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Through the comparisons among 91 tRNA Trp sequences from prokaryotes, archea and eukaryotes, the potential species-specific identity elements of tRNA Trp are found to be located within acceptor stem, dihydrouridine (D) stem, anticodon(AC) stem and discriminator base. Mutagenesis of B. subtilis tRNA Trp to the eukaryotic consensus se quence, in vitro transcription and enzymatic assay of tRNA Trp toward different tryptophanyl-tRNA synthetases (TrpRS) were employed to shed light on these species-specific identity elements and demonstrate the accurate recognition and the coevolution between tRNA and TrpRS. B. subtilis tRNA Trp with its acceptor stem and discriminator base transplanted by eukaryotic counterparts exhibited diminished reactivity toward B. subtilis enzyme but could be efficiently aminoacylated by rat liver crude enzyme. In contrast, B. subtilis tRNA Trp analog with the eukaryotic anticodon stem and D stem retains its recognition by B. subtilis enzyme. The results provide a strong evidence that the species-specific identity elements of tRNA Trp are orientated within the acceptor stem and discriminator base of tRNA Trp, and the anticodon stem and D stem are of little importance to the interaction between tRNA Trp and its cognate synthetase (TrpRS).

  10. Analytical Modeling of Surface Roughness, Hardness and Residual Stress Induced by Deep Rolling

    Science.gov (United States)

    Magalhães, Frederico C.; Abrão, Alexandre M.; Denkena, Berend; Breidenstein, Bernd; Mörke, Tobias

    2016-12-01

    Deep rolling is a mechanical surface treatment that can significantly alter the features of metallic components and despite the fact that it has been used for a long time, to date the influence of the interaction among the principal process parameters has not been thoroughly understood. Aiming to fulfill this gap, this work addresses the effect of deep rolling on surface finish and mechanical properties from the analytical and experimental viewpoints. More specifically, the influence of deep rolling pressure and number of passes on surface roughness, hardness and residual stress induced on AISI 1060 steel is investigated. The findings indicate that the surface roughness after deep rolling is closely related to the yield strength of the work material and the available models can satisfactorily predict the former parameter. Better agreement between the mathematical and experimental hardness values is achieved when a single deep rolling pass is employed, as well as when the yield strength of the work material increases. Compressive residual stress is generally induced after deep rolling, irrespectively of the selected heat treatment and deep rolling parameters. Finally, the model proposed to predict residual stress provides results closest to the experimental data especially when the annealed material is considered.

  11. RESIDUAL EFFECT OF LAMBDACYHALOTHRIN (ICON 10% WP ON DIFFERENT SURFACES IN SOUTH OF IRAN

    Directory of Open Access Journals (Sweden)

    H. Ladonni

    1994-08-01

    Full Text Available An investigation was made to study the residual activity and air-borne effect of lambdacyhalothrin (Icon 10 WP at 30, 40 and 50 a.i. mg/rn2 against An.stephensi in hut scale trials on different surfaces in Mamasani, and Rudan, south of Iran. Bio-assay contact mortality test with Icon at 30 a.i. mgl/m2 on plaster, cement and wood surfaces decreased from 100% to 51.4%, 53% and 63.8% after 84, 80 and 107 days of hut spraying, respectively. At 40 a.i mg/m2 Icon showed longer residual activity than the 30 mg/rn2, i.e. the mortality rate deceased from 100% to 50.7% for plaster, to 52.5% for cement and to 53.5% for wood surfaces, after 105, 105 and 124 days of hut spraying, respectively. Icon at 50 a.i. mg/m2 showed longer residual activity than the former dosages, i.e. the mortality dropped from 100% to 62.1% for plaster, to 50.7% for cement and to 90.4% for wooden surfaces, after 124 days of hut spraying, respectively. The result of air home tests showed that Icon has negligible or slight killing effect on An.stephensi, i.e. at 30. 40 and 50 a.i. mg/m2, the mortality rates dropped from 100% to about 60% after 10, 15 and 25 days of hut spraying, respectively. Based on the results of this study lambdacyhalothrin at 50/m2 had a residual activity for more than 4 months on different surface. It could be concluded that, lambdacyhalothrin at 50 mg/m2 might be a candidate dosage for continuation of study in village scale vial in south of Iran.

  12. pH-induced structural change of a multi-tryptophan protein MPT63 with immunoglobulin-like fold: identification of perturbed tryptophan residue/residues.

    Science.gov (United States)

    Mukherjee, Manini; Ghosh, Ranendu; Chattopadhyay, Krishnananda; Ghosh, Sanjib

    2015-01-01

    The structural change of M. tuberculosis MPT63, which is predominantly a β-sheet protein having an immunoglobulin like fold, has been investigated in the pH range 7.5-1.5 using various biophysical techniques along with low-temperature phosphorescence (LTP) spectroscopy. MPT63 contains four Tryptophan (Trp) residues at 26, 48, 82, and 129. Although circular dichroism, steady-state and time-resolved fluorescence, time-resolved anisotropy, 1-aniline-8-naphthalene sulfonic (ANS) acid binding, and analytical ultracentrifuge depict more open largely unfolded structure of MPT63 at pH 1.5 and also more accessible nature of Trp residues to neutral quencher at pH 1.5, it is, however, not possible to assign the specific Trp residue/residues being perturbed. This problem has been resolved using LTP of MPT63, which shows optically resolved four distinct (0, 0) bands corresponding to four Trp residues in the pH range 7.5-3.0. LTP at pH 1.5 clearly reveals that the solvent-exposed Trp 82 and the almost buried Trp 129 are specifically affected compared with Trp 48 and Trp 26. Lys 8 and Lys 27 are predicted to affect Trp 129. Tyrosine residues are found to be silent even at pH 1.5. This type of specific perturbation in a multi-Trp protein has not been addressed before. LTP further indicates that partially exposed Trp 48 is preferentially quenched by acrylamide compared with other Trp residues at both pH 7.5 and 1.5. The solvent-exposed Trp 82 is surprisingly found to be not quenched by acrylamide at pH 7.5. All the results are obtained using micromolar concentration of protein and without using any Trp-substituted mutant.

  13. Effect of Laser Shock Peening on surface properties and residual stress of Al6061-T6

    Science.gov (United States)

    Salimianrizi, A.; Foroozmehr, E.; Badrossamay, M.; Farrokhpour, H.

    2016-02-01

    The purpose of this study is to investigate the effects of Laser Shock Peening (LSP) on Al 6061-T6. The confined LSP regime using Nd: YAG laser with 1200 mJ of energy per pulse and 8 ns of pulse width were applied. The treated specimens were evaluated by means of surface integrity with optical microscopy, scanning electron microscope, microhardness, surface roughness and induced residual stress using an X-ray diffraction method. Results showed that by the use of LSP, compressive residual stress could effectively be induced on the surface of treated material. It was also revealed that the hardened depth of the material, up to a maximum depth of 1875 μm, could be achieved due to work hardening and grain refinement. In addition, surface roughness measurements showed that the LSP could deteriorate surface quality depending on the LSP parameters. The influences of beam overlap rates, number of laser shots and scanning pattern on microhardness as well as surface roughness are discussed.

  14. Residual stress and electromagnetic characteristics in loop type frequency selective surface embedded hybrid structures

    Energy Technology Data Exchange (ETDEWEB)

    Park, Kyung Mi; Seo, Yun Seok; Chun, Heoung Jae [Yonsei University, Seoul (Korea, Republic of); Hong, Ik Pyo [Kongju National University, Cheonan (Korea, Republic of); Park, Yong Bae [Ajoo University, Suwon (Korea, Republic of); Kim, Yoon Jae [Agency for defense development, Daejeon (Korea, Republic of)

    2015-01-15

    Residual stresses occur in frequency-selective surface (FSS)-embedded composite structures after co-curing due to differences between the coefficients of thermal expansion between composite skins and FSSs. Furthermore, the electromagnetic characteristics may be affected by the deformation of the FSS pattern by residual stresses. Therefore, we studied the changes in electromagnetic characteristics due to the deformation of FSS, using residual stresses to deform loop-type FSS-embedded hybrid composites. We considered the effects of loop-type FSS patterns of equal dimension as well as the stacking sequences of composite laminates on the electromagnetic characteristics of FSSs: Square loop, triangular loop and circular loop. The stacking sequences of composite laminates considered in this study were [0]{sub 8}, [0/90]{sub 4}, [+-45]{sub 4} and [0/+-45/90]{sub 2}. The FSS was located between composite laminates in the middle plane. To determine the residual stresses and deformations in the FSS embedded laminate structures, the thermal loading condition in the finite element analysis was induced by cooling the hybrid structures from 125 .deg. C to 20 .deg. C based on the cure cycle of the composite. Also, the electromagnetic reflection characteristics of the hybrid structures were predicted using deformed models by residual stresses, considering the effects of stacking sequence of composite laminates. The results showed that the maximum residual stresses and deformations were produced in the [0]{sub 8} composites with all three loop-types of FSS pattern. However, the maximum resonance frequency shifts occurred in the square and triangle loop-types with stacking sequence of [0]{sub 8} , while the maximum resonance frequency shift occurred in the circular loop-type with stacking sequence of [0/+-45/90]{sub 2}.

  15. Residual Stress on Surface and Cross-section of Porous Silicon Studied by Micro-Raman Spectroscopy

    Institute of Scientific and Technical Information of China (English)

    LEI Zhen-Kun; KANG YI-Lan; CEN Hao; HU Ming; QIU Yu

    2005-01-01

    @@ Surface and cross-sectional residual stresses of electrochemical etching porous silicon are investigated quantitatively by micro-Raman spectroscopy.The results reveal that a larger tensile residual stress exists on the surface and increase linearly with the porosity.On the other hand, across the depth direction perpendicular to the surface, the tensile residual stress decreases gradually from the surface to regions near the interface between the porous silicon layer and the Si substrate.However, a compressive stress appears at the interface near to the Si substrate for balancing with the tensile stress in the porous silicon layer.The cross-sectional residual stress profile is due to the porosity and lattice mismatch gradients existing in the cross-section and influencing each other.Furthermore, the presented residual stresses of the porous silicon have a close relation with its microstructure.

  16. Sequence composition and environment effects on residue fluctuations in protein structures

    Science.gov (United States)

    Ruvinsky, Anatoly M.; Vakser, Ilya A.

    2010-10-01

    Structure fluctuations in proteins affect a broad range of cell phenomena, including stability of proteins and their fragments, allosteric transitions, and energy transfer. This study presents a statistical-thermodynamic analysis of relationship between the sequence composition and the distribution of residue fluctuations in protein-protein complexes. A one-node-per-residue elastic network model accounting for the nonhomogeneous protein mass distribution and the interatomic interactions through the renormalized inter-residue potential is developed. Two factors, a protein mass distribution and a residue environment, were found to determine the scale of residue fluctuations. Surface residues undergo larger fluctuations than core residues in agreement with experimental observations. Ranking residues over the normalized scale of fluctuations yields a distinct classification of amino acids into three groups: (i) highly fluctuating-Gly, Ala, Ser, Pro, and Asp, (ii) moderately fluctuating-Thr, Asn, Gln, Lys, Glu, Arg, Val, and Cys, and (iii) weakly fluctuating-Ile, Leu, Met, Phe, Tyr, Trp, and His. The structural instability in proteins possibly relates to the high content of the highly fluctuating residues and a deficiency of the weakly fluctuating residues in irregular secondary structure elements (loops), chameleon sequences, and disordered proteins. Strong correlation between residue fluctuations and the sequence composition of protein loops supports this hypothesis. Comparing fluctuations of binding site residues (interface residues) with other surface residues shows that, on average, the interface is more rigid than the rest of the protein surface and Gly, Ala, Ser, Cys, Leu, and Trp have a propensity to form more stable docking patches on the interface. The findings have broad implications for understanding mechanisms of protein association and stability of protein structures.

  17. The persistence of illicit drug smoke residues and their recovery from common household surfaces.

    Science.gov (United States)

    Bitter, Julie L

    2017-04-01

    Third-hand smoke is the residue remaining on surfaces during smoking events. It is composed of particles and vapours that form upon heating. The phrase 'third-hand smoke' is primarily used to describe nicotine and other chemicals from cigarettes, but any residues formed from the smoking of various substances could be classified similarly. There has been an increasing body of research on third-hand smoke from cigarettes in the last decade, but little has been done in regards to understanding the persistence of particles and vapours from illicit drugs. In this work, small samples of cocaine and methamphetamine were volatilized to produce an illicit drug smoke that was collected onto various surface materials and left exposed to ambient conditions over 672 h (four weeks). Chemical analyses by electrospray ionization-mass spectrometry of residues on silicon, plastic, laminate, and artificial leather surfaces indicated a rapid decrease in recovery of the parent molecule, with varied formation of decomposition products over the first 168 h of exposure. Measurable amounts of the parent molecule were still present after 672 h, exhibiting a strong persistence of these drugs on various household materials. This is important in a forensic science context, as third-hand smoke residues could provide a viable source of trace evidence previously not utilized. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  18. Corroding and Protecting of Surface Residual Stress With the Surface Hardening of Steel

    Institute of Scientific and Technical Information of China (English)

    ZOU Qing-hua

    2004-01-01

    The states of the stress with the surface hardening are analyzed, the related experiential formula of the stress -temperature and stress-depth hardened curves are set up, the protecting measures against hydrogen stress corrosion are discussed.

  19. Residues of endosulfan in surface and subsurface agricultural soil and its bioremediation.

    Science.gov (United States)

    Odukkathil, Greeshma; Vasudevan, Namasivayam

    2016-01-01

    The persistence of many hydrophobic pesticides has been reported by various workers in various soil environments and its bioremediation is a major concern due to less bioavailability. In the present study, the pesticide residues in the surface and subsurface soil in an area of intense agricultural activity in Pakkam Village of Thiruvallur District, Tamilnadu, India, and its bioremediation using a novel bacterial consortium was investigated. Surface (0-15 cm) and subsurface soils (15-30 cm and 30-40 cm) were sampled, and pesticides in different layers of the soil were analyzed. Alpha endosulfan and beta endosulfan concentrations ranged from 1.42 to 3.4 mg/g and 1.28-3.1 mg/g in the surface soil, 0.6-1.4 mg/g and 0.3-0.6 mg/g in the subsurface soil (15-30 cm), and 0.9-1.5 mg/g and 0.34-1.3 mg/g in the subsurface soil (30-40 cm) respectively. Residues of other persistent pesticides were also detected in minor concentrations. These soil layers were subjected to bioremediation using a novel bacterial consortium under a simulated soil profile condition in a soil reactor. The complete removal of alpha and beta endosulfan was observed over 25 days. Residues of endosulfate were also detected during bioremediation, which was subsequently degraded on the 30th day. This study revealed the existence of endosulfan in the surface and subsurface soils and also proved that the removal of such a ubiquitous pesticide in the surface and subsurface environment can be achieved in the field by bioaugumenting a biosurfactant-producing bacterial consortium that degrades pesticides.

  20. Residual compressive surface stress increases the bending strength of dental zirconia.

    Science.gov (United States)

    Inokoshi, Masanao; Zhang, Fei; Vanmeensel, Kim; De Munck, Jan; Minakuchi, Shunsuke; Naert, Ignace; Vleugels, Jozef; Van Meerbeek, Bart

    2017-04-01

    To assess the influence of surface treatment and thermal annealing on the four-point bending strength of two ground dental zirconia grades. Fully-sintered zirconia specimens (4.0×3.0×45.0mm(3)) of Y-TZP zirconia (LAVA Plus, 3M ESPE) and Y-TZP/Al2O3 zirconia (ZirTough, Kuraray Noritake) were subjected to four surface treatments: (1) 'GROUND': all surfaces were ground with a diamond-coated grinding wheel on a grinding machine; (2) 'GROUND+HEAT': (1) followed by annealing at 1100°C for 30min; (3) 'GROUND+Al2O3 SANDBLASTED': (1) followed by sandblasting using Al2O3; (4) 'GROUND+CoJet SANDBLASTED': (1) followed by tribochemical silica (CoJet) sandblasting. Micro-Raman spectroscopy was used to assess the zirconia-phase composition and potentially induced residual stress. The four-point bending strength was measured using a universal material-testing machine. Weibull analysis revealed a substantially higher Weibull modulus and slightly higher characteristic strength for ZirTough (Kuraray Noritake) than for LAVA Plus (3M ESPE). For both zirconia grades, the 'GROUND' zirconia had the lowest Weibull modulus in combination with a high characteristic strength. Sandblasting hardly changed the bending strength but substantially increased the Weibull modulus of the ground zirconia, whereas a thermal treatment increased the Weibull modulus of both zirconia grades but resulted in a significantly lower bending strength. Micro-Raman analysis revealed a higher residual compressive surface stress that correlated with an increased bending strength. Residual compressive surface stress increased the bending strength of dental zirconia. Thermal annealing substantially reduced the bending strength but increased the consistency (reliability) of 'GROUND' zirconia. Copyright © 2017 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  1. A wafer mapping technique for residual stress in surface micromachined films

    Science.gov (United States)

    Schiavone, G.; Murray, J.; Smith, S.; Desmulliez, M. P. Y.; Mount, A. R.; Walton, A. J.

    2016-09-01

    The design of MEMS devices employing movable structures is crucially dependant on the mechanical behaviour of the deposited materials. It is therefore important to be able to fully characterize the micromachined films and predict with confidence the mechanical properties of patterned structures. This paper presents a characterization technique that enables the residual stress in MEMS films to be mapped at the wafer level by using microstructures released by surface micromachining. These dedicated MEMS test structures and the associated measurement techniques are used to extract localized information on the strain and Young’s modulus of the film under investigation. The residual stress is then determined by numerically coupling this data with a finite element analysis of the structure. This paper illustrates the measurement routine and demonstrates it with a case study using electrochemically deposited alloys of nickel and iron, particularly prone to develop high levels of residual stress. The results show that the technique enables wafer mapping of film non-uniformities and identifies wafer-to-wafer differences. A comparison between the results obtained from the mapping technique and conventional wafer bow measurements highlights the benefits of using a procedure tailored to films that are non-uniform, patterned and surface-micromachined, as opposed to simple standard stress extraction methods. The presented technique reveals detailed information that is generally unexplored when using conventional stress extraction methods such as wafer bow measurements.

  2. Quantitative assessments of residual stress fields at the surface of alumina hip joints.

    Science.gov (United States)

    Pezzotti, Giuseppe; Munisso, Maria Chiara; Lessnau, Kristina; Zhu, Wenliang

    2010-11-01

    In-depth and in-plane response functions of photo- and electro-stimulated probes have been modeled and quantitatively evaluated in order to assess their suitability to detect the highly graded residual stress fields generated at the surface of alumina hip joints. Optical calibrations revealed large differences in probe size, which strongly affected the detected magnitude of residual stress. A comparison between the responses of Raman and fluorescence probes in polycrystalline alumina showed that the depth of those probes spread to an extent in the order of the tens of microns even with using a confocal probe configuration. On the other hand, the electro-stimulated luminescence emitted by oxygen vacancy sites (F(+) center) in the alumina lattice represented the most suitable choice for confining to a shallow volume the stress probe. This latter probe enabled us to reduce the measurement depth to the order of the tens of nanometers. We show maps of surface residual stress as collected on both main-wear and nonwear zones of an alumina femoral head. A comparison among stress maps taken at exactly the same location, but employing different probes, revealed averaging effects on the stress magnitude detected with photo-stimulated probes, while proving the superior spatial resolution of the electron probe.

  3. Anti-tumor Immunity Elicited by Adenovirus Encoding AdhTrp2 or AdmTrp2 without Vitiligo

    Institute of Scientific and Technical Information of China (English)

    Hongju LIU; Xianzhi XIONG; Zuoya LI; Jianbao XIN; Xiaonan TAO; Yu HU

    2008-01-01

    To compare the difference in tumor immunity and autoimmunity elicited by adenovirus (Ad) encoding human or murine tyrosinase-related protein 2 (AdhTRP2 or AdmTRP2), and to find the most effective way to induce immunity by AdhTRP2 or AdmTRP2, C57BL/6 mice were im-munized with AdhTRP2 or AdmTRP2 intramuscularly at different doses of 105, 106, 107 and 108 separately (10 mice for each dose). Two weeks after the immunization, in vivo CTL assay and in- tracellular staining (ICS) of IFN-γ were carried out to analyze the dose-effect relationship. Tumor growth and vitiligo (as an sign of autoimmunity) were observed until 3 months after challenge with 105 B I6F10 tumor cells. The results showed that Ad encoding AdmTrp2 induced weak tumor im- mune response. Similar immunization with AdhTrp-2 elicited stronger protective immunity. CTL activity and IFN-γ-produced CD8+T cells were directly proportional to dose of AdhTrp2 or AdmTrp2. Moreover, AdhTrp2 group showed tumor rejection in 100% of challenged mice till the end of 3rd month while 60% of mice immunized with AdmTrp2 were protected against tumor. In the whole process of this experiment, no vitiligo was observed in mice immunized either with AdhTrp2 or AdmTrp2. It is concluded that anti-melanoma responses induced by genetic vaccina- tion expressing xenoantigens breaks immune tolerance effectively and is able to elicit strong anti-gen-specific cytotoxic T cell response without vitiligo.

  4. Surface Residual Stresses in Ti-6Al-4V Friction Stir Welds: Pre- and Post-Thermal Stress Relief

    Science.gov (United States)

    Edwards, P.; Ramulu, M.

    2015-09-01

    The purpose of this study was to determine the residual stresses present in titanium friction stir welds and if a post-weld thermal stress relief cycle would be effective in minimizing those weld-induced residual stresses. Surface residual stresses in titanium 6Al-4V alloy friction stir welds were measured in butt joint thicknesses ranging from 3 to 12 mm. The residual stress states were also evaluated after the welds were subjected to a post-weld thermal stress relief cycle of 760 °C for 45 min. High (300-400 MPa) tensile residual stresses were observed in the longitudinal direction prior to stress relief and compressive residual stresses were measured in the transverse direction. After stress relief, the residual stresses were decreased by an order of magnitude to negligible levels.

  5. Requiring pollutant discharge permits for pesticide applications that deposit residues in surface waters.

    Science.gov (United States)

    Centner, Terence; Eberhart, Nicholas

    2014-05-08

    Agricultural producers and public health authorities apply pesticides to control pests that damage crops and carry diseases. Due to the toxic nature of most pesticides, they are regulated by governments. Regulatory provisions require pesticides to be registered and restrictions operate to safeguard human health and the environment. Yet pesticides used near surface waters pose dangers to non-target species and drinking water supplies leading some governments to regulate discharges of pesticides under pollution discharge permits. The dual registration and discharge permitting provisions are burdensome. In the United States, agricultural interest groups are advancing new legislation that would exempt pesticide residues from water permitting requirements. An analysis of the dangers posed by pesticide residues in drinking water leads to a conclusion that both pesticide registration and pollutant discharge permitting provisions are needed to protect human health and aquatic species.

  6. Requiring Pollutant Discharge Permits for Pesticide Applications that Deposit Residues in Surface Waters

    Directory of Open Access Journals (Sweden)

    Terence Centner

    2014-05-01

    Full Text Available Agricultural producers and public health authorities apply pesticides to control pests that damage crops and carry diseases. Due to the toxic nature of most pesticides, they are regulated by governments. Regulatory provisions require pesticides to be registered and restrictions operate to safeguard human health and the environment. Yet pesticides used near surface waters pose dangers to non-target species and drinking water supplies leading some governments to regulate discharges of pesticides under pollution discharge permits. The dual registration and discharge permitting provisions are burdensome. In the United States, agricultural interest groups are advancing new legislation that would exempt pesticide residues from water permitting requirements. An analysis of the dangers posed by pesticide residues in drinking water leads to a conclusion that both pesticide registration and pollutant discharge permitting provisions are needed to protect human health and aquatic species.

  7. Mutations in Transhydrogenase Change the Fluorescence Emission State of TRP72 from 1La to 1Lb

    Science.gov (United States)

    Tveen Jensen, Karina; Strambini, Giovanni; Gonnelli, Margherita; Broos, Jaap; Jackson, J. Baz

    2008-01-01

    The dI component of Rhodospirillum rubrum transhydrogenase has a single Trp residue (Trp72), which has distinctive optical properties, including short-wavelength fluorescence emission with clear vibrational fine structure, and long-lived, well-resolved phosphorescence emission. We have made a set of mutant dI proteins in which residues contacting Trp72 are conservatively substituted. The room-temperature fluorescence-emission spectra of our three Met97 mutants are blue shifted by ∼4 nm, giving them a shorter-wavelength emission than any other protein described in the literature, including azurin from Pseudomonas aeruginosa. Fluorescence spectra in low-temperature glasses show equivalent well-resolved vibrational bands in wild-type and the mutant dI proteins, and in azurin. Substitution of Met97 in dI changes the relative intensities of some of these vibrational bands. The analysis supports the view that fluorescence from the Met97 mutants arises predominantly from the 1Lb excited singlet state of Trp72, whereas 1La is the predominant emitting state in wild-type dI. It is suggested that the sulfur atom of Met97 promotes greater stabilization of 1La than either 1Lb or the ground state. The phosphorescence spectra of Met97 mutants are also blue-shifted, indicating that the sulfur atom decreases the transition energy between the 3La state of the Trp and the ground state. PMID:18599622

  8. X-ray measurement of residual stresses in laser surface melted Ti-6Al-4V alloy

    NARCIS (Netherlands)

    Robinson, J.M.; van Brussel, B.A.; de Hosson, J.T.M.; Reed, R.C.

    1996-01-01

    In this paper, we report on the residual stresses in laser surface melted Ti-6Al-4V, determined using X-ray diffraction methods. The principal result is that there is an increase in the transverse residual stress with each successive, overlapping laser track. The result can be used to explain the ob

  9. Influence of Surface Properties and Impact Conditions on Insect Residue Adhesion

    Science.gov (United States)

    Wohl, Christopher J.; Doss, Jereme R.; Shanahan, Michelle H.; Smith, Joseph G., Jr.; Penner, Ronald K.; Connell, John W.; Siochi, Emilie J.

    2015-01-01

    Airflow over airfoils used on current commercial aircraft transitions from laminar to turbulent at relatively low chord positions. As a result, drag increases, requiring more thrust to maintain flight. An airfoil with increased laminar flow would experience reduced drag and a lower fuel burn rate. One of the objectives of NASA's Environmentally Responsible Aviation project is to identify and demonstrate technologies that will enable more environmentally friendly commercial aircraft. While more aerodynamically efficient airfoil shapes can be designed, surface contamination from ice, dirt, pollen, runway debris, and insect residue can degrade performance.

  10. Inequivalent contribution of the five tryptophan residues in the C-lobe of human serum transferrin to the fluorescence increase when iron is released.

    Science.gov (United States)

    James, Nicholas G; Byrne, Shaina L; Steere, Ashley N; Smith, Valerie C; MacGillivray, Ross T A; Mason, Anne B

    2009-04-07

    Human serum transferrin (hTF), with two Fe3+ binding lobes, transports iron into cells. Diferric hTF preferentially binds to a specific receptor (TFR) on the surface of cells, and the complex undergoes clathrin dependent receptor-mediated endocytosis. The clathrin-coated vesicle fuses with an endosome where the pH is lowered, facilitating iron release from hTF. On a biologically relevant time scale (2-3 min), the factors critical to iron release include pH, anions, a chelator, and the interaction of hTF with the TFR. Previous work, in which the increase in the intrinsic fluorescence signal was used to monitor iron release from the hTF/TFR complex, established that the TFR significantly enhances the rate of iron release from the C-lobe of hTF. In the current study, the role of the five C-lobe Trp residues in reporting the fluorescence change has been evaluated (+/-sTFR). Only four of the five recombinant Trp --> Phe mutants produced well. A single slow rate constant for iron release is found for the monoferric C-lobe (FeC hTF) and the four Trp mutants in the FeC hTF background. The three Trp residues equivalent to those in the N-lobe differed from the N-lobe and each other in their contributions to the fluorescent signal. Two rate constants are observed for the FeC hTF control and the four Trp mutants in complex with the TFR: k(obsC1) reports conformational changes in the C-lobe initiated by the TFR, and k(obsC2) is ascribed to iron release. Excitation at 295 nm (Trp only) and at 280 nm (Trp and Tyr) reveals interesting and significant differences in the rate constants for the complex.

  11. On-matrix Derivatization for Dynamic Headspace Sampling of Nonvolatile Surface Residues

    Energy Technology Data Exchange (ETDEWEB)

    Harvey, Scott D.; Wahl, Jon H.

    2012-09-01

    The goal of this study is to extend sampling by the field and laboratory emission cell (FLEC) purge-and-trap technique to applications that target nonvolatile residues. On-matrix derivatization of residues to render analytes stable and more volatile is explored to achieve this goal. Results show that on-matrix derivatizations of nerve agent hydrolysis products (monoalkyl methylphosphonic acids and methylphosphonic acid [MPA]) with diazomethane were successful on glass and painted wallboard (at the 10-µg level). It also was successful on the more difficult concrete (at the 500-µg level) and carpet (at the 20-µg level) substrates that cannot be successfully sampled using swipe techniques. Analysis of additional chemical warfare (CW)-associated residues can be approached by on-matrix derivatization with trifluoroacetic anhydride (TFAA). For example, amines (used as stabilizers or present as decomposition products of the nerve agent VX) or thiodiglycol (hydrolysis product of sulfur mustard) could be sampled as their TFAA derivatives from glass, painted wallboard, and concrete (at the 40-µg level), as well as carpet (at the 80-µg level) surfaces. Although the amine and thiodiglycol are semi-volatile and could be sampled directly, derivatization improves the recovery and chromatographic behavior of these analytes.

  12. On-matrix derivatization for dynamic headspace sampling of nonvolatile surface residues.

    Science.gov (United States)

    Harvey, Scott D; Wahl, Jon H

    2012-09-21

    The goal of this study is to extend sampling by the field and laboratory emission cell (FLEC) dynamic headspace technique to applications that target nonvolatile residues. On-matrix derivatization of residues to render analytes stable and more volatile is explored to achieve this goal. Results show that on-matrix derivatizations of nerve agent hydrolysis products (monoalkyl methylphosphonic acids and methylphosphonic acid [MPA]) with diazomethane were successful on glass and painted wallboard (at the 10-μg level). It also was successful on the more difficult concrete (at the 500-μg level) and carpet (at the 20-μg level), substrates that cannot be successfully sampled using swipe techniques. Analysis of additional chemical warfare (CW)-associated residues can be approached by on-matrix derivatization with trifluoroacetic anhydride (TFAA). For example, amines (used as stabilizers or present as decomposition products of the nerve agent VX) or thiodiglycol (hydrolysis product of sulfur mustard) could be sampled as their TFAA derivatives from glass, painted wallboard, and concrete (at the 40-μg level), as well as carpet (at the 80-μg level) surfaces. Although the amine and thiodiglycol are semi-volatile and could be sampled directly, derivatization improves the recovery and chromatographic behavior of these analytes.

  13. The evaluation of hierarchical structured superhydrophobic coatings for the alleviation of insect residue to aircraft laminar flow surfaces

    Science.gov (United States)

    Kok, Mariana; Young, Trevor M.

    2014-09-01

    Surface contamination caused by insects on laminar flow wing surfaces causes a disruption of the flow, resulting in an increase in drag and fuel consumption. Consequently, the use of superhydrophobic coatings to mitigate insect residue adhesion was investigated. A range of hierarchical superhydrophobic coatings with different surface chemistry and topography was examined. Candidate coatings were characterized in terms of their morphology and hydrophobic properties by scanning electron microscopy (SEM) and static and dynamic contact angle measurements, respectively. Arithmetic mean surface roughness (Ra) values were measured using profilometry. Only superhydrophobic coatings with a specific topography showed complete mitigation against insect residue adhesion. A surface which exhibited a specific microstructure (Ra = 5.26 μm) combined with a low sliding angle (SA = 7.6°) showed the best anti-contamination properties. The dynamics of an insect impact event and its influence on the wetting and adhesion mechanisms of insect residue to a surface were discussed.

  14. The evaluation of hierarchical structured superhydrophobic coatings for the alleviation of insect residue to aircraft laminar flow surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Kok, Mariana [Department of Mechanical, Aeronautical and Biomedical Engineering, University of Limerick, Limerick (Ireland); Materials and Surface Science Institute, University of Limerick, Limerick (Ireland); Young, Trevor M., E-mail: Trevor.Young@ul.ie [Department of Mechanical, Aeronautical and Biomedical Engineering, University of Limerick, Limerick (Ireland); Materials and Surface Science Institute, University of Limerick, Limerick (Ireland)

    2014-09-30

    Surface contamination caused by insects on laminar flow wing surfaces causes a disruption of the flow, resulting in an increase in drag and fuel consumption. Consequently, the use of superhydrophobic coatings to mitigate insect residue adhesion was investigated. A range of hierarchical superhydrophobic coatings with different surface chemistry and topography was examined. Candidate coatings were characterized in terms of their morphology and hydrophobic properties by scanning electron microscopy (SEM) and static and dynamic contact angle measurements, respectively. Arithmetic mean surface roughness (R{sub a}) values were measured using profilometry. Only superhydrophobic coatings with a specific topography showed complete mitigation against insect residue adhesion. A surface which exhibited a specific microstructure (R{sub a} = 5.26 μm) combined with a low sliding angle (SA = 7.6°) showed the best anti-contamination properties. The dynamics of an insect impact event and its influence on the wetting and adhesion mechanisms of insect residue to a surface were discussed.

  15. Noncontact dipole effects on channel permeation. I. Experiments with (5F-indole)Trp13 gramicidin A channels.

    OpenAIRE

    Busath, D D; Thulin, C D; Hendershot, R W; Phillips, L R; Maughan, P; Cole, C D; Bingham, N C; Morrison, S.; Baird, L C; Hendershot, R J; Cotten, M; Cross, T A

    1998-01-01

    Gramicidin A (gA), with four Trp residues per monomer, has an increased conductance compared to its Phe replacement analogs. When the dipole moment of the Trp13 side chain is increased by fluorination at indole position 5 (FgA), the conductance is expected to increase further. gA and FgA conductances to Na+, K+, and H+ were measured in planar diphytanoylphosphatidylcholine (DPhPC) or glycerylmonoolein (GMO) bilayers. In DPhPC bilayers, Na+ and K+ conductances increased upon fluorination, wher...

  16. Variations in Surface Texture of the North Polar Residual Cap of Mars

    Science.gov (United States)

    Milkovich, S. M.; Byrne, S.; Russell, P. S.

    2011-01-01

    The northern polar residual cap (NPRC) of Mars is a water ice deposit with a rough surface made up of pits, knobs, and linear depressions on scales of tens of meters. This roughness manifests as a series of bright mounds and dark hollows in visible images; these bright and dark patches have a characteristic wavelength and orientation. Spectral data indicate that the surface of the NPRC is composed of large-grained (and therefore old) water ice. Due to the presence of this old ice, it is thought that the NPRC is in a current state of net loss of material a result potentially at odds with impact crater statistics, which suggest ongoing deposition over the past 10-20 Kyr.

  17. Variations in Surface Texture of the North Polar Residual Cap of Mars

    Science.gov (United States)

    Milkovich, S. M.; Byrne, S.; Russell, P. S.

    2011-01-01

    The northern polar residual cap (NPRC) of Mars is a water ice deposit with a rough surface made up of pits, knobs, and linear depressions on scales of tens of meters. This roughness manifests as a series of bright mounds and dark hollows in visible images; these bright and dark patches have a characteristic wavelength and orientation. Spectral data indicate that the surface of the NPRC is composed of large-grained (and therefore old) water ice. Due to the presence of this old ice, it is thought that the NPRC is in a current state of net loss of material a result potentially at odds with impact crater statistics, which suggest ongoing deposition over the past 10-20 Kyr.

  18. Residual Viral and Bacterial Contamination of Surfaces after Cleaning and Disinfection

    Science.gov (United States)

    Tuladhar, Era; Hazeleger, Wilma C.; Koopmans, Marion; Zwietering, Marcel H.; Beumer, Rijkelt R.

    2012-01-01

    Environmental surfaces contaminated with pathogens can be sources of indirect transmission, and cleaning and disinfection are common interventions focused on reducing contamination levels. We determined the efficacy of cleaning and disinfection procedures for reducing contamination by noroviruses, rotavirus, poliovirus, parechovirus, adenovirus, influenza virus, Staphylococcus aureus, and Salmonella enterica from artificially contaminated stainless steel surfaces. After a single wipe with water, liquid soap, or 250-ppm free chlorine solution, the numbers of infective viruses and bacteria were reduced by 1 log10 for poliovirus and close to 4 log10 for influenza virus. There was no significant difference in residual contamination levels after wiping with water, liquid soap, or 250-ppm chlorine solution. When a single wipe with liquid soap was followed by a second wipe using 250- or 1,000-ppm chlorine, an extra 1- to 3-log10 reduction was achieved, and except for rotavirus and norovirus genogroup I, no significant additional effect of 1,000 ppm compared to 250 ppm was found. A reduced correlation between reduction in PCR units (PCRU) and reduction in infectious particles suggests that at least part of the reduction achieved in the second step is due to inactivation instead of removal alone. We used data on infectious doses and transfer efficiencies to estimate a target level to which the residual contamination should be reduced and found that a single wipe with liquid soap followed by a wipe with 250-ppm free chlorine solution was sufficient to reduce the residual contamination to below the target level for most of the pathogens tested. PMID:22941071

  19. Method for the Collection, Gravimetric and Chemical Analysis of Nonvolatile Residue (NVR) on Surfaces

    Science.gov (United States)

    Gordon, Keith; Rutherford, Gugu; Aranda, Denisse

    2017-01-01

    Nonvolatile residue (NVR), sometimes referred to as molecular contamination is the term used for the total composition of the inorganic and high boiling point organic components in particulates and molecular films deposited on critical surfaces surrounding space structures, with the particulate and NVR contamination originating primarily from pre-launch operations. The "nonvolatile" suggestion from the terminology NVR implies that the collected residue will not experience much loss under ambient conditions. NVR has been shown to have a dramatic impact on the ability to perform optical measurements from platforms based in space. Such contaminants can be detected early by the controlled application of various detection techniques and contamination analyses. Contamination analyses are the techniques used to determine if materials, components, and subsystems can be expected to meet the performance requirements of a system. Of particular concern is the quantity of NVR contaminants that might be deposited on critical payload surfaces from these sources. Subsequent chemical analysis of the contaminant samples by infrared spectroscopy and gas chromatography mass spectrometry identifies the components, gives semi-quantitative estimates of contaminant thickness, indicates possible sources of the NVR, and provides guidance for effective cleanup procedures. In this report, a method for the collection and determination of the mass of NVR was generated by the authors at NASA Langley Research Center. This report describes the method developed and implemented for collecting NVR contaminants, and procedures for gravimetric and chemical analysis of the residue obtained. The result of this NVR analysis collaboration will help pave the way for Langley's ability to certify flight hardware outgassing requirements in support of flight projects such as Stratospheric Aerosol and Gas Experiment III (SAGE III), Clouds and the Earth's Radiant Energy System (CERES), Materials International

  20. Exploiting Oceanic Residual Depth to Quantify Present-day Dynamic Topography at the Earth's Surface

    Science.gov (United States)

    Hoggard, Mark; White, Nicky

    2014-05-01

    Convective circulation within the mantle causes vertical motions at the Earth's surface. This dynamic topography is time dependent and occurs on wavelengths of 1000s km with maximum amplitudes of ±2 km. Convective simulation models have been used extensively to make predictions of dynamic topography and have thus far out-paced observational constraints. Here, the well-established relationship between seafloor subsidence and age is used to produce a global map of residual depth anomalies in the oceanic realm. Care is taken to remove other causes of topography, including an isostatic correction for sedimentary loading that takes compaction into account, a correction for variable oceanic crustal thickness, and lithospheric thickening with age away from mid-ocean ridge spreading centres. A dataset including over 1000 seismic reflection profiles and 300 modern wide-angle refraction experiments has been amassed, primarily on old ocean floor adjacent to the continents. Calculation of residual depth yields a map of present-day dynamic topography with amplitudes significantly larger than the errors associated with the corrections. One of the most interesting results occurs along the west coast of Africa, where two full 2000 km wavelengths of dynamic topography have been captured with amplitudes ±1 km that correlate well with the long-wavelength free air gravity anomaly. Comparison with predictive models reveal poor to moderate correlations. This is a direct result of the limited resolution of the mantle tomography models used to set-up convection simulations and also the currently poor understanding of viscosity structure within the Earth. It is hoped that this residual depth dataset should provide an excellent surface boundary constraint for future convective simulation.

  1. An enhanced trend surface analysis equation for regional-residual separation of gravity data

    Science.gov (United States)

    Obasi, A. I.; Onwuemesi, A. G.; Romanus, O. M.

    2016-12-01

    Trend surface analysis is a geological term for a mathematical technique which separates a given map set into a regional component and a local component. This work has extended the steps for the derivation of the constants in the trend surface analysis equation from the popularly known matrix and simultaneous form to a more simplified and easily achievable format. To achieve this, matrix inversion was applied to the existing equations and the outcome was tested for suitability using a large volume of gravity data set acquired from the Anambra Basin, south-eastern Nigeria. Tabulation of the field data set was done using the Microsoft Excel spread sheet, while gravity maps were generated from the data set using Oasis Montaj software. A comparison of the residual gravity map produced using the new equations with its software derived counterpart has shown that the former has a higher enhancing capacity than the latter. This equation has shown strong suitability for application in the separation of gravity data sets into their regional and residual components.

  2. Portable surface plasmon resonance immunosensor for the detection of fluoroquinolone antibiotic residues in milk.

    Science.gov (United States)

    Fernández, Fátima; Pinacho, Daniel G; Sánchez-Baeza, Francisco; Marco, M Pilar

    2011-05-11

    An inexpensive and portable surface plasmon resonance (SPR) sensor, SPReeta Evaluation Kit SPR3, has been used to develop a biosensor for the determination of fluoroquinolone antibiotics (FQs) and to demonstrate its performance analyzing FQ residues in milk samples. The SPReeta three-channel gold chips were activated with a mixed self-assembled monolayer (m-SAM) and functionalized with a FQ haptenized protein. Binding of the antibody produced a concentration-dependent increase of the SPR signal as a result of the change in the refraction index. Similarly, the presence of the FQ produced a dose-dependent decrease of the response, which allowed a good limit of detection (LOD) to be obtained (1.0 ± 0.4 μg L(-1) for enrofloxacin in buffer). The response was reproducible in all three channels, on different injections and days, and also between chips. Milk samples could be analyzed after a simple sample treatment involving fat removal by centrifugation and dilution with water. Under these conditions calibration curves were obtained showing that FQ residues can be analyzed in milk samples with an IC(50) value of 26.4 ± 7.2 μg L(-1) and a LOD of 2.0 ± 0.2 μg L(-1) (for enrofloxacin), far below the European Union regulations for this antibiotic family in this matrix. Finally, the paper also demonstrates that the biosensor is able to selectively detect the presence of FQs in milk samples, even in the presence of other antibiotics. Enrofloxacin, ciprofloxacin, and norfloxacin residues were detected in blind samples supplied by Nestlé Co.

  3. Instrumental aspects of TPD / TRP techniques; Aspetti strumentali delle tecniche TPD / TRP

    Energy Technology Data Exchange (ETDEWEB)

    Dossi, C.; Recchia, S. [CNR, Milan (Italy). Dipt. di Chimica Inorganica, Metallorganica e Analitica

    1997-11-01

    Criteria for designing the most suitable experimental setup for TPD / TRP measurements are discussed in detail. Interfacing the control and measurement components with a PC represents a crucial step in this operation. Two different approaches are considered: the first based on a computer-linked gas chromatographic integrator, the second featuring an analogic/ digital conversion module. When complex probe molecules or chemical reactions are involved, combining selective detectors such as GC and MS is recommended as the best solution.

  4. Comparison pesticide residue levels in the surface of Bertam River in Cameron Highlands, Pahang

    Energy Technology Data Exchange (ETDEWEB)

    Haron, S. H., E-mail: ismail@ukm.edu.my; Ismail, B. S., E-mail: sthumaira@yahoo.com [School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia 43600 UKM, Bangi, Selangor (Malaysia)

    2015-09-25

    The presence of pesticide residues in the surface water of Bertam River in the agricultural areas of Cameron Highlands in Pahang, Malaysia was monitored from May to October 2014. The sampling sites were located at 10 sampling points along the Bertam River in the vegetable planting areas. The extraction method of the pesticide (organophosphate/pyrethroid) from the river samples used solid phase extraction followed by gas chromatography (with electron capture detector, ECD). Insecticides, cypermethrin and chlorpyrifos were found in the surface water of Bertam River. High level concentrations of those insecticides in the river were observed during the period from May to October 2014, a period which included both seasons (wet and dry seasons). The highest concentration of 2.66 µg/mL and 1.23 µg/mL of cypermethrin was observed during the wet and dry seasons respectively. This could be due to the frequent usage of the above-mentioned insecticides coupled with contamination that could have originated from the application sites. Meanwhile, the lowest concentration detected in the surface water was chlorpyrifos (0.11 µg/mL and 0.17 µg/mL) during the dry and wet seasons, respectively.

  5. Comparison pesticide residue levels in the surface of Bertam River in Cameron Highlands, Pahang

    Science.gov (United States)

    Haron, S. H.; Ismail B., S.

    2015-09-01

    The presence of pesticide residues in the surface water of Bertam River in the agricultural areas of Cameron Highlands in Pahang, Malaysia was monitored from May to October 2014. The sampling sites were located at 10 sampling points along the Bertam River in the vegetable planting areas. The extraction method of the pesticide (organophosphate/pyrethroid) from the river samples used solid phase extraction followed by gas chromatography (with electron capture detector, ECD). Insecticides, cypermethrin and chlorpyrifos were found in the surface water of Bertam River. High level concentrations of those insecticides in the river were observed during the period from May to October 2014, a period which included both seasons (wet and dry seasons). The highest concentration of 2.66 µg/mL and 1.23 µg/mL of cypermethrin was observed during the wet and dry seasons respectively. This could be due to the frequent usage of the above-mentioned insecticides coupled with contamination that could have originated from the application sites. Meanwhile, the lowest concentration detected in the surface water was chlorpyrifos (0.11 µg/mL and 0.17 µg/mL) during the dry and wet seasons, respectively.

  6. Identifying Surface Changes on HRSC Images of the Mars South Polar Residual CAP (sprc)

    Science.gov (United States)

    Putri, Alfiah Rizky Diana; Sidiropoulos, Panagiotis; Muller, Jan-Peter

    2016-06-01

    The surface of Mars has been an object of interest for planetary research since the launch of Mariner 4 in 1964. Since then different cameras such as the Viking Visual Imaging Subsystem (VIS), Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC), and Mars Reconnaissance Orbiter (MRO) Context Camera (CTX) and High Resolution Imaging Science Experiment (HiRISE) have been imaging its surface at ever higher resolution. The High Resolution Stereo Camera (HRSC) on board of the European Space Agency (ESA) Mars Express, has been imaging the Martian surface, since 25th December 2003 until the present-day. HRSC has covered 100 % of the surface of Mars, about 70 % of the surface with panchromatic images at 10-20 m/pixel, and about 98 % at better than 100 m/pixel (Neukum et. al., 2004), including the polar regions of Mars. The Mars polar regions have been studied intensively recently by analysing images taken by the Mars Express and MRO missions (Plaut et al., 2007). The South Polar Residual Cap (SPRC) does not change very much in volume overall but there are numerous examples of dynamic phenomena associated with seasonal changes in the atmosphere. In particular, we can examine the time variation of layers of solid carbon dioxide and water ice with dust deposition (Bibring, 2004), spider-like channels (Piqueux et al., 2003) and so-called Swiss Cheese Terrain (Titus et al., 2004). Because of seasonal changes each Martian year, due to the sublimation and deposition of water and CO2 ice on the Martian south polar region, clearly identifiable surface changes occur in otherwise permanently icy region. In this research, good quality HRSC images of the Mars South Polar region are processed based on previous identification as the optimal coverage of clear surfaces (Campbell et al., 2015). HRSC images of the Martian South Pole are categorized in terms of quality, time, and location to find overlapping areas, processed into high quality Digital Terrain Models (DTMs) and

  7. IDENTIFYING SURFACE CHANGES ON HRSC IMAGES OF THE MARS SOUTH POLAR RESIDUAL CAP (SPRC

    Directory of Open Access Journals (Sweden)

    A. R. D. Putri

    2016-06-01

    Full Text Available The surface of Mars has been an object of interest for planetary research since the launch of Mariner 4 in 1964. Since then different cameras such as the Viking Visual Imaging Subsystem (VIS, Mars Global Surveyor (MGS Mars Orbiter Camera (MOC, and Mars Reconnaissance Orbiter (MRO Context Camera (CTX and High Resolution Imaging Science Experiment (HiRISE have been imaging its surface at ever higher resolution. The High Resolution Stereo Camera (HRSC on board of the European Space Agency (ESA Mars Express, has been imaging the Martian surface, since 25th December 2003 until the present-day. HRSC has covered 100 % of the surface of Mars, about 70 % of the surface with panchromatic images at 10-20 m/pixel, and about 98 % at better than 100 m/pixel (Neukum et. al., 2004, including the polar regions of Mars. The Mars polar regions have been studied intensively recently by analysing images taken by the Mars Express and MRO missions (Plaut et al., 2007. The South Polar Residual Cap (SPRC does not change very much in volume overall but there are numerous examples of dynamic phenomena associated with seasonal changes in the atmosphere. In particular, we can examine the time variation of layers of solid carbon dioxide and water ice with dust deposition (Bibring, 2004, spider-like channels (Piqueux et al., 2003 and so-called Swiss Cheese Terrain (Titus et al., 2004. Because of seasonal changes each Martian year, due to the sublimation and deposition of water and CO2 ice on the Martian south polar region, clearly identifiable surface changes occur in otherwise permanently icy region. In this research, good quality HRSC images of the Mars South Polar region are processed based on previous identification as the optimal coverage of clear surfaces (Campbell et al., 2015. HRSC images of the Martian South Pole are categorized in terms of quality, time, and location to find overlapping areas, processed into high quality Digital Terrain Models (DTMs and

  8. REPLACEMENT OF TRYPTOPHAN RESIDUES IN HALOALKANE DEHALOGENASE REDUCES HALIDE BINDING AND CATALYTIC ACTIVITY

    NARCIS (Netherlands)

    KENNES, C; PRIES, F; KROOSHOF, GH; BOKMA, E; Kingma, Jacob; JANSSEN, DB

    1995-01-01

    Haloalkane dehalogenase catalyzes the hydrolytic cleavage of carbon-halogen bonds in short-chain haloalkanes. Two tryptophan residues of the enzyme (Trp125 and Trp175) form a halide-binding site in the active-site cavity, and were proposed to play a role in catalysis. The function of these residues

  9. Two-dimensional fluorescence correlation spectroscopy IV: Resolution of fluorescence of tryptophan residues in alcohol dehydrogenase and lysozyme

    Science.gov (United States)

    Fukuma, Hiroaki; Nakashima, Kenichi; Ozaki, Yukihiro; Noda, Isao

    2006-11-01

    Generalized two-dimensional (2D) fluorescence correlation spectroscopy has been used to resolve the fluorescence spectra of two tryptophan (Trp) residues in alcohol dehydrogenase and lysozyme. In each protein, one Trp residue is buried in a hydrophobic domain of the protein matrix and the other Trp residue is located at a hydrophilic domain close to the protein-water interface. Fluorescence quenching by iodide ion, a hydrophilic quencher, was employed as a perturbation to induce the intensity change in the spectra. The Trp residue which is located at the hydrophilic domain is effectively quenched by the quencher, while the Trp residue located at the hydrophobic domain is protected from the quenching. Therefore, the fluorescence of these two Trp residues have a different sensitivity to the quenching, showing a different response to the concentration of the quencher. Fluorescence spectra of the two Trp residues in alcohol dehydrogenase, which are heavily overlapped in conventional one-dimensional spectra, have been successfully resolved by the 2D correlation technique. From the asynchronous correlation map, it was revealed that the quenching of Trp located at the hydrophobic part was brought about after that of Trp located at the hydrophilic part. In contrast, the fluorescence spectra of the two Trp residues could not be resolved after the alcohol dehydrogenase was denatured with guanidine hydrochloride. These results are consistent with the well-known structure of alcohol dehydrogenase. Furthermore, it was elucidated that the present 2D analysis is not interfered by Raman bands of the solvent, which sometimes bring difficulty into the conventional fluorescence analysis. Fluorescence spectra of the Trp residues in lysozyme could not be resolved by the 2D correlation technique. The differences between the two proteins are attributed to the fact that the Trp residue in the hydrophobic site of lysozyme is not sufficiently protected from the quenching.

  10. Trp63 is regulated by STAT5 in mammary tissue and subject to differentiation in cancer

    OpenAIRE

    Assefnia, Shahin; Kang, Keunsoo; Groeneveld, Svenja; Yamaji, Daisuke; Dabydeen, Sarah; Alamri, Ahmad; Liu, Xuefeng; Hennighausen, Lothar; Furth, Priscilla A

    2014-01-01

    Trp63, founding member of the Trp53 family, contributes to epithelial differentiation and is expressed in breast neoplasia. Trp63 features two distinct promoters yielding specific mRNAs encoding two major TRP63 isoforms, a transactivating transcription factor and a dominant negative isoform. Specific TRP63 isoforms are linked to cell cycle arrest, apoptosis, survival and epithelial mesenchymal transition. Although TRP63 overexpression in cultured cells is used to elucidate functions, little i...

  11. Quantifying residual stress in nanoscale thin polymer films via surface wrinkling.

    Science.gov (United States)

    Chung, Jun Young; Chastek, Thomas Q; Fasolka, Michael J; Ro, Hyun Wook; Stafford, Christopher M

    2009-04-28

    Residual stress, a pervasive consequence of solid materials processing, is stress that remains in a material after external forces have been removed. In polymeric materials, residual stress results from processes, such as film formation, that force and then trap polymer chains into nonequilibrium stressed conformations. In solvent-cast films, which are central to a wide range of technologies, residual stress can cause detrimental effects, including microscopic defect formation and macroscopic dimensional changes. Since residual stress is difficult to measure accurately, particularly in nanoscale thin polymer films, it remains a challenge to understand and control. We present here a quantitative method of assessing residual stress in polymer thin films by monitoring the onset of strain-induced wrinkling instabilities. Using this approach, we show that thin (>100 nm) polystyrene films prepared via spin-coating possess residual stresses of approximately 30 MPa, close to the crazing and yield stress. In contrast to conventional stress measurement techniques such as wafer curvature, our technique has the resolution to measure residual stress in films as thin as 25 nm. Furthermore, we measure the dissipation of residual stress through two relaxation mechanisms: thermal annealing and plasticizer addition. In quantifying the amount of residual stress in these films, we find that the residual stress gradually decreases with increasing annealing time and plasticizer amounts. Our robust and simple route to measure residual stress adds a key component to the understanding of polymer thin film behavior and will enable identification of more effective processing routes that mitigate the detrimental effects of residual stress.

  12. Adsorptive removal of residual catalyst from palm biodiesel: Application of response surface methodology

    Directory of Open Access Journals (Sweden)

    Mjalli Sabri Farouq

    2012-01-01

    Full Text Available In this work, the residual potassium hydroxide catalyst was removed from palm oil-based methyl esters using an adsorption technique. The produced biodiesel was initially purified through a water washing process. To produce a biodiesel with a better quality and also to meet standard specifications (EN 14214 and ASTM D6751, batch adsorption on palm shell activated carbon was used for further catalyst removal. The Central Composite Design (CCD of the Response Surface Methodology (RSM was used to study the influence of adsorbent amount, time and temperature on the adsorption of potassium species. The maximum catalyst removal was achieved at 40°C using 0.9 g activated carbon for 20 h adsorption time. The results from the Response Surface Methodology are in a good agreement with the measured values. The absolute error in prediction at the optimum condition was 3.7%, which is reasonably accurate. This study proves that adsorption post-treatment techniques can be successfully employed to improve the quality of biodiesel fuel for its effective use on diesel engines and to minimize the usage of water.

  13. Large surface radiative forcing from topographic blowing snow residuals measured in the High Arctic at Eureka

    Directory of Open Access Journals (Sweden)

    G. Lesins

    2009-03-01

    Full Text Available Ice crystals, also known as diamond dust, are suspended in the boundary layer air under clear sky conditions during most of the Arctic winter in Northern Canada. Occasionally ice crystal events can produce significantly thick layers with optical depths in excess of 2.0 even in the absence of liquid water clouds. Four case studies of high optical depth ice crystal events at Eureka in the Nunavut Territory of Canada during the winter of 2006/07 are presented. They show that the measured ice crystal surface infrared downward radiative forcing ranged from 8 to 36 W m−2 in the wavelength band from 5.6 to 20 μm for 532 nm optical depths ranging from 0.2 to 1.7. MODIS infrared and visible images and the operational radiosonde wind profile were used to show that these high optical depth events were caused by surface snow being blown off 600 to 800 m high mountain ridges about 20 to 30 km North-West of Eureka and advected by the winds towards Eureka as they settled towards the ground within the highly stable boundary layer. This work presents the first study that demonstrates the important role that surrounding topography plays in determining the occurrence of high optical depth ice crystal events from residual blowing snow that becomes a source of boundary layer ice crystals distinct from the classical diamond dust phenomenon.

  14. Direct surface plasmon resonance immunosensing of pyraclostrobin residues in untreated fruit juices.

    Science.gov (United States)

    Mauriz, E; García-Fernández, C; Mercader, J V; Abad-Fuentes, A; Escuela, A M; Lechuga, L M

    2012-12-01

    A surface plasmon resonance (SPR) immunoassay for on-line detection of the strobilurin fungicide pyraclostrobin in untreated fruit juices is presented. The analysis of pyraclostrobin residues is accomplished in apple, grape, and cranberry samples by monitoring the recognition events occurring separately in a two-channel home-made SPR biosensor. Covalent coupling of the analyte derivative results in a reversible method, enabling more than 80 measurements on the same sensor surface. Optimization of the immunoassay conditions provides limits of detection as low as 0.16 μg L(-1). The selectivity and reproducibility of the analysis is ensured by studying both non-specific interactions with unrelated compounds and inter-assay coefficients of variation. Excellent recovery ranging from 98 to 103% was achieved by a simple 1:5 dilution of fruit juice with assay buffer before the analysis. The lack of previous cleaning and homogenization procedures reduces the analysis time of a single food sample to only 25 min, including the regeneration cycle.

  15. Residues and Characteristics of Organochlorine Pesticides in the Surface Water in the Suburb of Beijing

    Science.gov (United States)

    CHEN, Jiawei; LIU, Chen; YANG, Zhongfang; WANG, Jiyuan

    Organochlorine Pesticides (OCPs), such as DDT and HCH, have stable chemical properties and less biodegradability. As a kind of persistent organic pollutants, they have high risk to the environment and human health. Although DDT and HCH have been prohibited in China since 1983, they are still found in some soil and water nowadays. Water resource is very important in natural environment and essential for agriculture. The existence of OCPs in some surface water in Beijing has been detected with different levels according to previous investigations. In recent years, many measures have been taken to control the pollution and to monitor the environment, and more attention has been paid to the status of surface water today. In this study, the water samples were collected from the Wenyu, Beiyun, Yanqing, Fangshan, Changping, and Shunyi Rivers in the suburb of Beijing, and the residues and characteristics of DDT and HCH were studied. The results showed that the contents of DDTs and HCHs were ND-13.98 ng/L and 3.87-146.42 ng/L, respectively. According to the indicators of the ratio values of (DDD+DDE)/DDT and α-HCH/γ-HCH, the source of pollution and its potential risk are also discussed in this article.

  16. [Aging effect on mechanical properties in fluid resin. (Part 3) Affection of residual monomer on the surface morphology after tensile test by microscopic observation (author's transl)].

    Science.gov (United States)

    Horiuchi, A

    1981-04-01

    Affection of residual monomer on the surface morphology after tensile test was observed by an scanning microscope. Evaporation or leaching of the monomer gave big influence on the morphology of pearls and matrix. The observation suggested that the residual monomer existed mainly in the matrix. When the residual monomer disappeared, the surface morphology did not change by tensile load. It is the most important point to get good denture with fluid resin that we could decrease the residual monomer as possible.

  17. A channelopathy mechanism revealed by direct calmodulin activation of TrpV4.

    Science.gov (United States)

    Loukin, Stephen H; Teng, Jinfeng; Kung, Ching

    2015-07-28

    Ca(2+)-calmodulin (CaM) regulates varieties of ion channels, including Transient Receptor Potential vanilloid subtype 4 (TrpV4). It has previously been proposed that internal Ca(2+) increases TrpV4 activity through Ca(2+)-CaM binding to a C-terminal Ca(2+)-CaM binding domain (CBD). We confirmed this model by directly presenting Ca(2+)-CaM protein to membrane patches excised from TrpV4-expressing oocytes. Over 50 TRPV4 mutations are now known to cause heritable skeletal dysplasia (SD) and other diseases in human. We have previously examined 14 SD alleles and found them to all have gain-of-function effects, with the gain of constitutive open probability paralleling disease severity. Among the 14 SD alleles examined, E797K and P799L are located immediate upstream of the CBD. They not only have increase basal activity, but, unlike the wild-type or other SD-mutant channels examined, they were greatly reduced in their response to Ca(2+)-CaM. Deleting a 10-residue upstream peptide (Δ795-804) that covers the two SD mutant sites resulted in strong constitutive activity and the complete lack of Ca(2+)-CaM response. We propose that the region immediately upstream of CBD is an autoinhibitory domain that maintains the closed state through electrostatic interactions, and adjacent detachable Ca(2+)-CaM binding to CBD sterically interferes with this autoinhibition. This work further supports the notion that TrpV4 mutations cause SD by constitutive leakage. However, the closed conformation is likely destabilized by various mutations by different mechanisms, including the permanent removal of an autoinhibition documented here.

  18. Trp(56) of rac1 specifies interaction with a subset of guanine nucleotide exchange factors.

    Science.gov (United States)

    Gao, Y; Xing, J; Streuli, M; Leto, T L; Zheng, Y

    2001-12-14

    Signaling specificity of Rho GTPase pathways is achieved in part by selective interaction between members of the Dbl family guanine nucleotide exchange factors (GEFs) and their Rho GTPase substrates. For example, Trio, GEF-H1, and Tiam1 are a subset of GEFs that specifically activate Rac1 but not the closely related Cdc42. The Rac1 specificity of these GEFs appears to be governed by Rac1-GEF binding interaction. To understand the detailed mechanism underlying the GEF specificity issue, we have analyzed a panel of chimeras made between Rac1 and Cdc42 and examined a series of point mutants of Rac1 made at the switch I, switch II, and beta(2)/beta(3) regions for their ability to interact with and to be activated by the GEFs. The results reveal that Rac1 residues of both the switch I and switch II regions are involved in GEF docking and GEF-mediated nucleotide disruption, because mutation of Asp(38), Asn(39), Gln(61), Tyr(64), or Arg(66)/Leu(67) into Ala results in the loss of GEF binding, whereas mutation at Tyr(32), Asp(65), or Leu(70)/Ser(71) leads to the loss of GEF catalysis while retaining the binding capability. The region between amino acids 53-72 of Rac1 is required for specific recognition and activation by the GEFs, and Trp(56) in beta(3) appears to be the critical determinant. Introduction of Trp(56) to Cdc42 renders it fully responsive to the Rac-specific GEF in vitro and in cells. Further, a polypeptide derived from the beta(3) region of Rac1 including the Trp(56) residue serves as a specific inhibitor for Rac1 interaction with the GEFs. Taken together, these results indicate that Trp(56) is the necessary and sufficient determinant of Rac1 for discrimination by the subset of Rac1-specific GEFs and suggest that a compound mimicking Trp(56) action could be explored as an interfering reagent specifically targeting Rac1 activation.

  19. Mutagenesis of tGCN5 core region reveals two critical surface residues F90 and R140

    Energy Technology Data Exchange (ETDEWEB)

    Mehta, Kinjal Rajesh; Chan, Yan M.; Lee, Man X.; Yang, Ching Yao; Voloshchuk, Natalya [Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, 6 MetroTech Center, Brooklyn, NY 11201 (United States); Montclare, Jin Kim, E-mail: jmontcla@poly.edu [Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, 6 MetroTech Center, Brooklyn, NY 11201 (United States); Department of Biochemistry, SUNY-Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203 (United States)

    2010-09-24

    Research highlights: {yields} Mutagenesis of the tGCN5 core region reveals two residues important for function. {yields} Developed a fluorescent lysate-based activity assay to assess mutants. {yields} Surface-exposed residues F90 and R140 of tGCN5 are critical for H3 acetylation. -- Abstract: Tetrahymena General Control Non-Derepressor 5 (tGCN5) is a critical regulator of gene transcription via acetylation of histones. Since the acetylation ability has been attributed to the 'core region', we perform mutagenesis of residues within the tGCN5 'core region' in order to identify those critical for function and stability. Residues that do not participate in catalysis are identified, mutated and characterized for activity, structure and thermodynamic stability. Variants I107V, Q114L, A121T and A130S maintain the acetylation function relative to wild-type tGCN5, while variants F90Y, F112R and R140H completely abolish function. Of the three non-functional variants, since F112 is mutated into a non-homologous charged residue, a loss in function is expected. However, the remaining two variants are mutated into homologous residues, suggesting that F90 and R140 are critical for the activity of tGCN5. While mutation to homologous residue maintains acetylation of histone H3 for the majority of the variants, the two surface-exposed residues, F90 and R140, appear to be essential for tGCN5 function, structure or stability.

  20. TRP Channels as Therapeutic Targets in Diabetes and Obesity

    Science.gov (United States)

    Zsombok, Andrea; Derbenev, Andrei V.

    2016-01-01

    During the last three to four decades the prevalence of obesity and diabetes mellitus has greatly increased worldwide, including in the United States. Both the short- and long-term forecasts predict serious consequences for the near future, and encourage the development of solutions for the prevention and management of obesity and diabetes mellitus. Transient receptor potential (TRP) channels were identified in tissues and organs important for the control of whole body metabolism. A variety of TRP channels has been shown to play a role in the regulation of hormone release, energy expenditure, pancreatic function, and neurotransmitter release in control, obese and/or diabetic conditions. Moreover, dietary supplementation of natural ligands of TRP channels has been shown to have potential beneficial effects in obese and diabetic conditions. These findings raised the interest and likelihood for potential drug development. In this mini-review, we discuss possibilities for better management of obesity and diabetes mellitus based on TRP-dependent mechanisms. PMID:27548188

  1. Swift residue-screening identifies key N-glycosylated asparagines sufficient for surface expression of neuroglycoprotein Lingo-1.

    Science.gov (United States)

    Zhong, Xiaotian; Pocas, Jennifer; Liu, Yan; Wu, Paul W; Mosyak, Lidia; Somers, Will; Kriz, Ron

    2009-03-18

    Advances in genomics and proteomics have generated the needs for the efficient identification of key residues for structure and function of target proteins. Here we report the utilization of a new residue-screening approach, which combines a mammalian high-throughput transient expression system with a PCR-based expression cassette, for the study of the post-translational modification. Applying this approach results in a quick identification of essential N-glycosylation sites of a heavily glycosylated neuroglycoprotein Lingo-1, which are sufficient for the support of its surface expression. These key N-glycosylated sites uniquely locate on the concave surface of the elongated arc-shape structure of the leucine-rich repeat domain. The swift residue-screening approach may provide a new strategy for structural and functional analysis.

  2. TRP channels in microglia: role in physiology and disease

    OpenAIRE

    2015-01-01

    Microglia are responsible for modulating the cellular environment in the nervous system, where they have neuroprotective effects. In these cells has been reported the expression of different ion channels, including those belonging to the transient receptor potential (TRP) family. TRP are nonselective cation channels that have six transmembrane domains and both N-terminus and C-terminus domain located in the cytoplasm. These proteins can be activated by mechanical, thermal and chemical stimuli...

  3. POCIS sampling in combination with ELISA: screening of sulfonamide residues in surface and waste waters.

    Science.gov (United States)

    Černoch, Ivo; Fránek, Milan; Diblíková, Iva; Hilscherová, Klára; Randák, Tomáš; Ocelka, Tomáš; Bláha, Luděk

    2012-01-01

    Sulfonamide antibiotics coming from both human and veterinary medicine are among the most common emerging pollutants in freshwater. The present paper shows the successful application of passive sampling using POCIS in combination with an immunochemical ELISA technique and HPLC/MS/MS analysis to study the distribution of sulfonamides in streams around small towns in the Czech Republic, as well as around a major agglomeration of the city of Brno, including its waste water treatment plant (WWTP). Results indicated the presence of sulfonamides at most studied sites with concentrations ranging from POCIS. Very high levels were detected in both the influent and effluent of the Brno WWTP with maxima > 8000 ng SMX per POCIS. All samplers collected down-stream of the studied towns and WWTPs clearly showed an increase in sulfonamide drug residues. Higher concentrations were determined in rivers at the city of Brno agglomeration. In agreement with other available studies, these findings indicate low efficiency of conventional WWTPs to eliminate polar pharmaceuticals such as sulfonamides. Good performance and correlation with the LC/MS results, as well as ease of use, indicate good potential for the immunochemical ELISA technique to become the screening tool for sulfonamide determination in surface waters including passive samplers.

  4. Apoptosis induced by (di-isopropyloxyphoryl-Trp)2-Lys-OCH3 in K562 and HeLa cells

    Indian Academy of Sciences (India)

    Feng Liu; Shi-Ying Liu; Ping Xu; Zhen-Hua Xie; Guo-Ping Cai; Yu-Yang Jiang

    2008-03-01

    According to the method used in our laboratory, our group synthesized (DIPP-Trp)2-Lys-OCH3. It inhibited the proliferation of K562 and HeLa cells in a dose- and time-dependent manner with an IC50 of 15.12 and 42.23 M, respectively. (DIPP-Trp)2-Lys-OCH3 induced a dose-dependent increase of the G2/M cell population in K562 cells, and S cell population in HeLa cells; the sub-G0 population increased dramatically in both cell lines as seen by PI staining experiments using a FACS Calibur Flow cytometer (BeckmanCoulter, USA). Phosphatidylserine could significantly translocate to the surface of the membrane in (DIPP-Trp)2-Lys-OCH3-treated K562 and HeLa cells. The increase of an early apoptotic population was observed in a dose-dependent manner by both annexin-FITC and PI staining. It was concluded that (DIPP-Trp)2-Lys-OCH3 not only induced cells to enter into apoptosis, but also affected the progress of the cell cycle. It may have arrested the K562 and HeLa cells in the G2/M, S phases, respectively. The apoptotic pathway was pulsed at this point, resulting in the treated cells entering into programmed cell death. (DIPP-Trp)2-Lys-OCH3 is a potential anticancer drug that intervenes in the signalling pathway.

  5. TRP channels: sensors and transducers of gasotransmitter signals

    Directory of Open Access Journals (Sweden)

    Nobuaki eTakahashi

    2012-08-01

    Full Text Available The transient receptor potential (trp gene superfamily encodes cation channels that act as multimodal sensors for a wide variety of stimuli from outside and inside the cell. Upon sensing, they transduce electrical and Ca2+ signals via their cation channel activities. These functional features of TRP channels allow the body to react and adapt to different forms of environmental changes. Indeed, members of one class of TRP channels have emerged as sensors of gaseous messenger molecules that control various cellular processes. Nitric oxide (NO, a vasoactive gaseous molecule, regulates TRP channels directly via cysteine S-nitrosylation or indirectly via cGMP/PKG-dependent phosphorylation. Recent studies have revealed that changes in the availability of molecular oxygen (O2 also control the activation of TRP channels. Anoxia induced by O2-glucose deprivation and severe hypoxia (1% O2 activates TRPM7 and TRPC6, respectively, whereas TRPA1 has recently been identified as a novel sensor of hyperoxia and mild hypoxia (15% O2 in vagal and sensory neurons. TRPA1 also detects other gaseous molecules such as hydrogen sulfide (H2S and carbon dioxide (CO2. In this review, we focus on how signaling by gaseous molecules is sensed and integrated by TRP channels.

  6. Structural insights into the evolution of a non-biological protein: importance of surface residues in protein fold optimization.

    Directory of Open Access Journals (Sweden)

    Matthew D Smith

    Full Text Available Phylogenetic profiling of amino acid substitution patterns in proteins has led many to conclude that most structural information is carried by interior core residues that are solvent inaccessible. This conclusion is based on the observation that buried residues generally tolerate only conserved sequence changes, while surface residues allow more diverse chemical substitutions. This notion is now changing as it has become apparent that both core and surface residues play important roles in protein folding and stability. Unfortunately, the ability to identify specific mutations that will lead to enhanced stability remains a challenging problem. Here we discuss two mutations that emerged from an in vitro selection experiment designed to improve the folding stability of a non-biological ATP binding protein. These mutations alter two solvent accessible residues, and dramatically enhance the expression, solubility, thermal stability, and ligand binding affinity of the protein. The significance of both mutations was investigated individually and together, and the X-ray crystal structures of the parent sequence and double mutant protein were solved to a resolution limit of 2.8 and 1.65 A, respectively. Comparative structural analysis of the evolved protein to proteins found in nature reveals that our non-biological protein evolved certain structural features shared by many thermophilic proteins. This experimental result suggests that protein fold optimization by in vitro selection offers a viable approach to generating stable variants of many naturally occurring proteins whose structures and functions are otherwise difficult to study.

  7. Validation of an HPLC–UV method for the determination of digoxin residues on the surface of manufacturing equipment

    Directory of Open Access Journals (Sweden)

    ZORAN B. TODOROVIĆ

    2009-09-01

    Full Text Available In the pharmaceutical industry, an important step consists in the removal of possible drug residues from the involved equipments and areas. The cleaning procedures must be validated and methods to determine trace amounts of drugs have, therefore, to be considered with special attention. An HPLC–UV method for the determination of digoxin residues on stainless steel surfaces was developed and validated in order to control a cleaning procedure. Cotton swabs, moistened with methanol were used to remove any residues of drugs from stainless steel surfaces, and give recoveries of 85.9, 85.2 and 78.7 % for three concentration levels. The precision of the results, reported as the relative standard deviation (RSD, were below 6.3 %. The method was validated over a concentration range of 0.05–12.5 µg mL-1. Low quantities of drug residues were determined by HPLC–UV using a Symmetry C18 column (150´4.6 mm, 5 µm at 20 °C with an acetonitrile–water (28:72, v/v mobile phase at a flow rate of 1.1 mL min-1, an injection volume of 100 µL and were detected at 220 nm. A simple, selective and sensitive HPLC–UV assay for the determination of digoxin residues on stainless steel was developed, validated and applied.

  8. Dependence of the residual surface resistance of SRF cavities on the cooling rate through $T_\\mathrm{c}$

    CERN Document Server

    Romanenko, A; Melnychuk, O; Sergatskov, D A

    2014-01-01

    We report a strong effect of the cooling rate through $T_\\mathrm{c}$ on the amount of trapped external magnetic flux in superconducting niobium cavities. The effect is similar for all surface treatments including electropolishing with and without 120$^\\circ$C baking, and nitrogen doping. Direct magnetic field measurements on the cavity walls show that the effect stems from changes in the flux trapping efficiency at different cooling rates. Slow cooling leads to almost complete flux trapping and higher residual resistance while fast cooling leads to the much more efficient flux expulsion and lower residual resistance.

  9. Effect of laser surface hardening on the microstructure, hardness and residual stresses of austempered ductile iron grades

    Energy Technology Data Exchange (ETDEWEB)

    Soriano, C., E-mail: csoriano@tekniker.es [Tekniker-IK4, Manufacturing Processes Department, Avda. Otaola 20, CP-20600, Eibar (Gipuzkoa) (Spain); Leunda, J.; Lambarri, J.; Garcia Navas, V.; Sanz, C. [Tekniker-IK4, Manufacturing Processes Department, Avda. Otaola 20, CP-20600, Eibar (Gipuzkoa) (Spain)

    2011-06-01

    A study of the laser surface hardening process of two austempered ductile iron grades, with different austempering treatments has been carried out. Hardening was performed with an infrared continuous wave Nd:YAG laser in cylindrical specimens. The microstructure of the laser hardened samples was investigated using an optical microscope, microhardness profiles were measured and surface and radial residual stresses were studied by an X-ray diffractometer. Similar results were achieved for both materials. A coarse martensite with retained austenite structure was found in the treated area, resulting in a wear resistant effective layer of 0.6 mm to 1 mm with a microhardness between 650 HV and 800 HV. Compressive residual stresses have been found at the hardened area being in agreement with the microhardness and microstructural variations observed. The achieved results point out that the laser surface hardening is a suitable method for improving the mechanical properties of austempered ductile irons.

  10. Expression of the wild-type and the Cys-/Trp-less alpha 3 beta 3 gamma complex of thermophilic F1-ATPase in Escherichia coli.

    Science.gov (United States)

    Matsui, T; Yoshida, M

    1995-09-12

    The alpha, beta and gamma subunits of F1-ATPase from thermophilic Bacillus PS3 were expressed in Escherichia coli cells simultaneously in large amounts. Most of the expressed subunits assembled into a form of alpha 3 beta 3 gamma complex in E. coli cells and this complex was easily purified to homogeneity. The recombinant alpha 3 beta 3 gamma complex thus obtained showed similar enzymatic properties to the alpha 3 beta 3 gamma complex obtained by in vitro reconstitution from individual subunits (Yokoyama, K. et al. (1989) J. Biol. Chem. 264, 21837-21841) except that the former had several-fold higher ATPase activity than the latter. Using this expression system, a mutant alpha 3 beta 3 gamma complex with no Trp and Cys was generated by replacing alpha Cys193 and alpha Trp463 with Ser and Phe, respectively. This mutant complex was functionally intact, indicating both residues are not essential for catalysis. The Cys-/Trp-less complex is a convenient 'second wild type' enzyme from which one can generate mutants with Trp (as a fluorescent probe) or Cys (as an acceptor of a variety of probes) at desired positions without concern for 'background' Trp and Cys residues.

  11. Massive glycosaminoglycan-dependent entry of Trp-containing cell-penetrating peptides induced by exogenous sphingomyelinase or cholesterol depletion.

    Science.gov (United States)

    Bechara, Chérine; Pallerla, Manjula; Burlina, Fabienne; Illien, Françoise; Cribier, Sophie; Sagan, Sandrine

    2015-02-01

    Among non-invasive cell delivery strategies, cell-penetrating peptide (CPP) vectors represent interesting new tools. To get fundamental knowledge about the still debated internalisation mechanisms of these peptides, we modified the membrane content of cells, typically by hydrolysis of sphingomyelin or depletion of cholesterol from the membrane outer leaflet. We quantified and visualised the effect of these viable cell surface treatments on the internalisation efficiency of different CPPs, among which the most studied Tat, R9, penetratin and analogues, that all carry the N-terminal biotin-Gly4 tag cargo. Under these cell membrane treatments, only penetratin and R6W3 underwent a massive glycosaminoglycan (GAG)-dependent entry in cells. Internalisation of the other peptides was only slightly increased, similarly in the absence or the presence of GAGs for R9, and only in the presence of GAGs for Tat and R6L3. Ceramide formation (or cholesterol depletion) is known to lead to the reorganisation of membrane lipid domains into larger platforms, which can serve as a trap and cluster receptors. These results show that GAG clustering, enhanced by formation of ceramide, is efficiently exploited by penetratin and R6W3, which contains Trp residues in their sequence but not Tat, R9 and R6L3. Hence, these data shed new lights on the differences in the internalisation mechanism and pathway of these peptides that are widely used in delivery of cargo molecules.

  12. Folding Dynamics of the Trp-Cage Miniprotein: Evidence for a Native-Like Intermediate from Combined Time-Resolved Vibrational Spectroscopy and Molecular Dynamics Simulations

    NARCIS (Netherlands)

    Meuzelaar, H.; Marino, K.A.; Huerta-Viga, A.; Panman, M.R.; Smeenk, L.E.J.; Kettelarij, A.J.; van Maarseveen, J.H.; Timmerman, P.; Bolhuis, P.G.; Woutersen, S.

    2013-01-01

    Trp-cage is a synthetic 20-residue miniprotein which folds rapidly and spontaneously to a well-defined globular structure more typical of larger proteins. Due to its small size and fast folding, it is an ideal model system for experimental and theoretical investigations of protein folding

  13. Psychiatric Disorders and TRP Channels: Focus on Psychotropic Drugs.

    Science.gov (United States)

    Nazıroğlu, Mustafa; Demirdaş, Arif

    2015-01-01

    Psychiatric and neurological disorders are mostly associated with the changes in neural calcium ion signaling pathways required for activity-triggered cellular events. One calcium channel family is the TRP cation channel family, which contains seven subfamilies. Results of recent papers have discovered that calcium ion influx through TRP channels is important. We discuss the latest advances in calcium ion influx through TRP channels in the etiology of psychiatric disorders. Activation of TRPC4, TRPC5, and TRPV1 cation channels in the etiology of psychiatric disorders such as anxiety, fear-associated responses, and depression modulate calcium ion influx. Evidence substantiates that anandamide and its analog (methanandamide) induce an anxiolytic-like effect via CB1 receptors and TRPV1 channels. Intracellular calcium influx induced by oxidative stress has an significant role in the etiology of bipolar disorders (BDs), and studies recently reported the important role of TRP channels such as TRPC3, TRPM2, and TRPV1 in converting oxidant or nitrogen radical signaling to cytosolic calcium ion homeostasis in BDs. The TRPV1 channel also plays a function in morphine tolerance and hyperalgesia. Among psychotropic drugs, amitriptyline and capsazepine seem to have protective effects on psychiatric disorders via the TRP channels. Some drugs such as cocaine and methamphetamine also seem to have an important role in alcohol addiction and substance abuse via activation of the TRPV1 channel. Thus, we explore the relationships between the etiology of psychiatric disorders and TRP channel-regulated mechanisms. Investigation of the TRP channels in psychiatric disorders holds the promise of the development of new drug treatments.

  14. Antibiotic resistance genes and residual antimicrobials in cattle feedlot surface soil

    Science.gov (United States)

    Antibiotic residues and resistant bacteria in cattle feedlot manure may impact antibiotic resistance in the environment. This study investigated common antimicrobials (tetracyclines and monensin) and associated resistance genes in cattle feedlot soils over time. Animal diets and other feedlot soil...

  15. Modelling the Effects of Surface Residual Stresses on Fatigue Behavior of PM Disk Alloys Project

    Data.gov (United States)

    National Aeronautics and Space Administration — A finite element based model will be developed and validated to capture the evolution of residual stresses and cold work at machined features of compressor and...

  16. Residual Effects of Deltamthrin WG 25% as a New Formulationon Different Surfaces against Anopheles stephensi, inSoutheastern Iran

    Directory of Open Access Journals (Sweden)

    A Raeisi

    2010-06-01

    Full Text Available Background: Indoor residual spraying (IRS is functioned as national interventions against malaria in southeastern foci of Iran and deltamethrin WP one of the insecticides have been used since past decade. In this study, the residual activity of the wettable granule (WG was studied on different surfaces in hut scale trial against Anopheles stephensi  in Iranshahr district, southeastern Iran.Methods: Three dosages of 25, 40 and 50 mg a.i./m2 of deltamethrin WG 25% formulation were applied on plaster, cement, mud, and wooden surfaces using Hudson® X-pert compression sprayer having 10 litters capacity.Results: The residual effects of deltamethrin WG 25% on different surfaces was assessed based on reduction of mortality An. stepehnsi from 100% to about 70%. At 25, 40 and 50 mg a.i./m2 the WG formulation of deltamethrin had a bioefficacy for about 2, 3 and 4 months respectively.Conclusion: There was an expectable fluctuation in mortality of An. stephensi at different sprayed surfaces as well as dosages. The proposed 50 mg/m2 WG is the longest activity for up to 4 months which needs to be applied for two spraying cycles per year at the climatically condition of southwestern Iran.

  17. Residual Effects of Deltamthrin WG 25% as a New Formulationon Different Surfaces against Anopheles stephensi, inSoutheastern Iran

    Directory of Open Access Journals (Sweden)

    A Raeisi

    2010-06-01

    Full Text Available "n "nBackground: Indoor residual spraying (IRS is functioned as national interventions against malaria in southeastern foci of Iran and deltamethrin WP one of the insecticides have been used since past decade. In this study, the residual activity of the wettable granule (WG was studied on different surfaces in hut scale trial against Anopheles stephensi  in Iranshahr district, southeastern Iran. "n "n "nMethods: Three dosages of 25, 40 and 50 mg a.i./m2 of deltamethrin WG 25% formulation were applied on plaster, cement, mud, and wooden surfaces using Hudson® X-pert compression sprayer having 10 litters capacity. "n "nResults: The residual effects of deltamethrin WG 25% on different surfaces was assessed based on reduction of mortality An. stepehnsi from 100% to about 70%. At 25, 40 and 50 mg a.i./m2 the WG formulation of deltamethrin had a bioefficacy for about 2, 3 and 4 months respectively. "nConclusion: There was an expectable fluctuation in mortality of An. stephensi at different sprayed surfaces as well as dosages. The proposed 50 mg/m2 WG is the longest activity for up to 4 months which needs to be applied for two spraying cycles per year at the climatically condition of southwestern Iran.

  18. Surface preparation for XRD residual stress measurements; Preparacao de superficie para medicao de tensoes residuais em soldagem por DRX

    Energy Technology Data Exchange (ETDEWEB)

    Mishchenko, Andrii; Oliveira, Bruno Jose de; Scotti, Americo, E-mail: asmwelder@gmail.com, E-mail: brunojoliveira7@gmail.com, E-mail: ascotti@mecanica.ufu.br [Universidade Federal de Uberlandia (UFU), MG (Brazil)

    2016-10-15

    A characteristic feature of the X Ray Diffraction (XRD) method for stress determination is that measurements occur at a thin surface layer. Steel sheets come with surfaces modified by lamination, cleaning (sandblasting, grinding) and even corrosion, which induce residual stresses or roughness inherent to the material. Therefore, surface preparation prior to the residual stress measurement is essential, although no standard procedure seems to be available. A general recommendation is to remove a thin layer so that only residual stresses related to the welding process will be measured. In this study, the use of portable electrolytic equipment was evaluated for mechanized surface material removal. Chemical compositions of electrolytic solutions and the influence of current on the removed material, removal time and temperature during the process were studied. As a result, a suitable chemical solution for electro etching of low carbon steel was developed and a set of “soft” parameters that allowed the removal of about 300 um in a reasonable time was found. Higher currents reduce the removal time, yet increasing the consumption of the solution and plate temperature (which could adversely alter the microstructure or generate thermal stresses). Furthermore, the influence of these parameters on the operability of the process was demonstrated. (author)

  19. Evolutionary conservation and changes in insect TRP channels

    Directory of Open Access Journals (Sweden)

    Tominaga Makoto

    2009-09-01

    Full Text Available Abstract Background TRP (Transient Receptor Potential channels respond to diverse stimuli and thus function as the primary integrators of varied sensory information. They are also activated by various compounds and secondary messengers to mediate cell-cell interactions as well as to detect changes in the local environment. Their physiological roles have been primarily characterized only in mice and fruit flies, and evolutionary studies are limited. To understand the evolution of insect TRP channels and the mechanisms of integrating sensory inputs in insects, we have identified and compared TRP channel genes in Drosophila melanogaster, Bombyx mori, Tribolium castaneum, Apis mellifera, Nasonia vitripennis, and Pediculus humanus genomes as part of genome sequencing efforts. Results All the insects examined have 2 TRPV, 1 TRPN, 1 TRPM, 3 TRPC, and 1 TRPML subfamily members, demonstrating that these channels have the ancient origins in insects. The common pattern also suggests that the mechanisms for detecting mechanical and visual stimuli and maintaining lysosomal functions may be evolutionarily well conserved in insects. However, a TRPP channel, the most ancient TRP channel, is missing in B. mori, A. mellifera, and N. vitripennis. Although P. humanus and D. melanogaster contain 4 TRPA subfamily members, the other insects have 5 TRPA subfamily members. T. castaneum, A. mellifera, and N. vitripennis contain TRPA5 channels, which have been specifically retained or gained in Coleoptera and Hymenoptera. Furthermore, TRPA1, which functions for thermotaxis in Drosophila, is missing in A. mellifera and N. vitripennis; however, they have other Hymenoptera-specific TRPA channels (AmHsTRPA and NvHsTRPA. NvHsTRPA expressed in HEK293 cells is activated by temperature increase, demonstrating that HsTRPAs function as novel thermal sensors in Hymenoptera. Conclusion The total number of insect TRP family members is 13-14, approximately half that of mammalian TRP

  20. Evolutionary conservation and changes in insect TRP channels.

    Science.gov (United States)

    Matsuura, Hironori; Sokabe, Takaaki; Kohno, Keigo; Tominaga, Makoto; Kadowaki, Tatsuhiko

    2009-09-10

    TRP (Transient Receptor Potential) channels respond to diverse stimuli and thus function as the primary integrators of varied sensory information. They are also activated by various compounds and secondary messengers to mediate cell-cell interactions as well as to detect changes in the local environment. Their physiological roles have been primarily characterized only in mice and fruit flies, and evolutionary studies are limited. To understand the evolution of insect TRP channels and the mechanisms of integrating sensory inputs in insects, we have identified and compared TRP channel genes in Drosophila melanogaster, Bombyx mori, Tribolium castaneum, Apis mellifera, Nasonia vitripennis, and Pediculus humanus genomes as part of genome sequencing efforts. All the insects examined have 2 TRPV, 1 TRPN, 1 TRPM, 3 TRPC, and 1 TRPML subfamily members, demonstrating that these channels have the ancient origins in insects. The common pattern also suggests that the mechanisms for detecting mechanical and visual stimuli and maintaining lysosomal functions may be evolutionarily well conserved in insects. However, a TRPP channel, the most ancient TRP channel, is missing in B. mori, A. mellifera, and N. vitripennis. Although P. humanus and D. melanogaster contain 4 TRPA subfamily members, the other insects have 5 TRPA subfamily members. T. castaneum, A. mellifera, and N. vitripennis contain TRPA5 channels, which have been specifically retained or gained in Coleoptera and Hymenoptera. Furthermore, TRPA1, which functions for thermotaxis in Drosophila, is missing in A. mellifera and N. vitripennis; however, they have other Hymenoptera-specific TRPA channels (AmHsTRPA and NvHsTRPA). NvHsTRPA expressed in HEK293 cells is activated by temperature increase, demonstrating that HsTRPAs function as novel thermal sensors in Hymenoptera. The total number of insect TRP family members is 13-14, approximately half that of mammalian TRP family members. As shown for mammalian TRP channels, this

  1. Expression and distribution of transient receptor potential (TRP) channels in bladder epithelium.

    Science.gov (United States)

    Yu, Weiqun; Hill, Warren G; Apodaca, Gerard; Zeidel, Mark L

    2011-01-01

    The urothelium is proposed to be a sensory tissue that responds to mechanical stress by undergoing dynamic membrane trafficking and neurotransmitter release; however, the molecular basis of this function is poorly understood. Transient receptor potential (TRP) channels are ideal candidates to fulfill such a role as they can sense changes in temperature, osmolarity, and mechanical stimuli, and several are reported to be expressed in the bladder epithelium. However, their complete expression profile is unknown and their cellular localization is largely undefined. We analyzed expression of all 33 TRP family members in mouse bladder and urothelium by RT-PCR and found 22 specifically expressed in the urothelium. Of the latter, 10 were chosen for closer investigation based on their known mechanosensory or membrane trafficking functions in other cell types. Western blots confirmed urothelial expression of TRPC1, TRPC4, TRPV1, TRPV2, TRPV4, TRPM4, TRPM7, TRPML1, and polycystins 1 and 2 (PKD1 and PKD2) proteins. We further defined the cellular and subcellular localization of all 10 TRP channels. TRPV2 and TRPM4 were prominently localized to the umbrella cell apical membrane, while TRPC4 and TRPV4 were identified on their abluminal surfaces. TRPC1, TRPM7, and TRPML1 were localized to the cytoplasm, while PKD1 and PKD2 were expressed on the apical and basolateral membranes of umbrella cells as well as in the cytoplasm. The cellular location of TRPV1 in the bladder has been debated, but colocalization with neuronal marker calcitonin gene-related peptide indicated clearly that it is present on afferent neurons that extend into the urothelium, but may not be expressed by the urothelium itself. These findings are consistent with the hypothesis that the urothelium acts as a sentinel and by expressing multiple TRP channels it is likely it can detect and presumably respond to a diversity of external stimuli and suggest that it plays an important role in urothelial signal

  2. TRP channels in microglia: role in physiology and disease

    Directory of Open Access Journals (Sweden)

    Yolima P Torres

    2015-02-01

    Full Text Available Microglia are responsible for modulating the cellular environment in the nervous system, where they have neuroprotective effects. In these cells has been reported the expression of different ion channels, including those belonging to the transient receptor potential (TRP family. TRP are nonselective cation channels that have six transmembrane domains and both N-terminus and C-terminus domain located in the cytoplasm. These proteins can be activated by mechanical, thermal and chemical stimuli as well as pH changes. Their activation can induce calcium influx contributing to regulate the intracellular calcium concentration. Different studies describe the effect of TRP channels in cellular processes such as osmotic regulation, cytokine production, proliferation, activation, cell death and responses to oxidative stress. As a result, these channels have an important role in neurodegenerative diseases, ischemia, inflammatory responses and pain perception. Modulating the activity of the TRP channels can be a therapeutic target for various diseases associated with the central nervous system. In this review the most relevant aspects about the role of the TRP channels in the physiology of microglia and their potential use as a therapeutic target in central nervous system diseases will be described.

  3. Functional and fluorescence analyses of tryptophan residues in H+-pyrophosphatase of Clostridium tetani.

    Science.gov (United States)

    Chen, Yen-Wei; Lee, Ching-Hung; Huang, Yun-Tzu; Pan, Yih-Jiuan; Lin, Shih-Ming; Lo, Yueh-Yu; Lee, Chien-Hsien; Huang, Lin-Kun; Huang, Yu-Fen; Hsu, Yu-Di; Pan, Rong-Long

    2014-04-01

    Homodimeric proton-translocating pyrophosphatase (H+-PPase; EC 3.6.1.1) maintains the cytoplasmic pH homeostasis of many bacteria and higher plants by coupling pyrophosphate (PPi) hydrolysis and proton translocation. H+-PPase accommodates several essential motifs involved in the catalytic mechanism, including the PPi binding motif and Acidic I and II motifs. In this study, 3 intrinsic tryptophan residues, Trp-75, Trp-365, and Trp-602, in H+-PPase from Clostridium tetani were used as internal probes to monitor the local conformational state of the periplasm domain, transmembrane region, and cytoplasmic domain, respectively. Upon binding of the substrate analog Mg-imidodiphosphate (Mg-IDP), local structural changes prevented the modification of tryptophan residues by N-bromosuccinimide (NBS), especially at Trp-602. Following Mg-Pi binding, Trp-75 and Trp-365, but not Trp-602, were slightly protected from structural modifications by NBS. These results reveal the conformation of H+-PPase is distinct in the presence of different ligands. Moreover, analyses of the Stern-Volmer relationship and steady-state fluorescence anisotropy also indicate that the local structure around Trp-602 is more exposed to solvent and varied under different environments. In addition, Trp-602 was identified to be a crucial residue in the H+-PPase that may potentially be involved in stabilizing the structure of the catalytic region by site-directed mutagenesis analysis.

  4. Performance Analysis of Flat Surface Assumption and Residual Motion Errors on Airborne Repeat-pass InSAR

    Directory of Open Access Journals (Sweden)

    Lin Xue

    2013-09-01

    Full Text Available When applying to the airborne repeat-pass Interferometric Synthetic Aperture Radar (InSAR, which has long synthetic aperture and large azimuth-dependent errors, the surface assumption used to simply the time-domain algorithm model and the residual motion errors due to the precision of the navigation system will affect the imaging result and the interferometric measurement. This paper analyzes the altitude errors introduced by the surface assumption and the residual motion errors due to the precision of the navigation system. We deduce the range errors model during the single pass and analyze the effects of these errors on the plane location, interferometric phase and DEM precision. Then the accuracy of the theoretical deduction is verified by simulation and real data. The research provides theoretical bases for the system design and signal processing of airborne repeat-pass InSAR.

  5. TRP ion channels in thermosensation, thermoregulation and metabolism

    Science.gov (United States)

    Wang, Hong; Siemens, Jan

    2015-01-01

    In humans, the TRP superfamily of cation channels includes 27 related molecules that respond to a remarkable variety of chemical and physical stimuli. While physiological roles for many TRP channels remain unknown, over the past years several have been shown to function as molecular sensors in organisms ranging from yeast to humans. In particular, TRP channels are now known to constitute important components of sensory systems, where they participate in the detection or transduction of osmotic, mechanical, thermal, or chemosensory stimuli. We here summarize our current understanding of the role individual members of this versatile receptor family play in thermosensation and thermoregulation, and also touch upon their immerging role in metabolic control. PMID:27227022

  6. Oxidation of free, peptide and protein tryptophan residues mediated by AAPH-derived free radicals: role of alkoxyl and peroxyl radicals

    DEFF Research Database (Denmark)

    Fuentes-Lemus, E.; Dorta, E.; Escobar, E.

    2016-01-01

    The oxidation of tryptophan (Trp) residues, mediated by peroxyl radicals (ROOc), follows a complex mechanism involving free radical intermediates, and short chain reactions. The reactivity of Trp towards ROOc should be strongly affected by its inclusion in peptides and proteins. To examine...... the latter, we investigated (by fluorescence) the kinetic of the consumption of free, peptide- and protein-Trp residues towards AAPH (2,20 -azobis(2-amidinopropane)dihydrochloride)-derived free radicals. Interestingly, the initial consumption rates (Ri ) were only slightly influenced by the inclusion of Trp...... concentrations (10–50 mM), the values of Ri were nearly constant; and at high Trp concentrations (50 mM to 1 mM), a slower increase of Ri than expected for chain reactions. Similar behavior was detected for all three systems (free Trp, and Trp in peptides and proteins). For the first time we are showing...

  7. Methodological considerations to understand the sensory function of TRP channels.

    Science.gov (United States)

    Meseguer, Víctor M; Denlinger, Bristol L; Talavera, Karel

    2011-01-01

    Transient Receptor Potential channels are exquisite molecular transducers of multiple physical and chemical stimuli, hence the raising interest to study their relevance to Sensory Biology. Here we discuss a number of aspects of the biophysical and pharmacological properties of TRP channels, which we consider essential for a clear understanding of their sensory function in vivo. By examining concrete examples extracted from recent literature we illustrate that TRP channel research is a field in motion, and that many established dogmas on biophysical properties, drug specificity and physiological role are continuously reshaped, and sometimes even dismantled.

  8. Sensory TRP channels: the key transducers of nociception and pain.

    Science.gov (United States)

    Mickle, Aaron D; Shepherd, Andrew J; Mohapatra, Durga P

    2015-01-01

    Peripheral detection of nociceptive and painful stimuli by sensory neurons involves a complex repertoire of molecular detectors and/or transducers on distinct subsets of nerve fibers. The majority of such molecular detectors/transducers belong to the transient receptor potential (TRP) family of cation channels, which comprise both specific receptors for distinct nociceptive stimuli, as well as for multiple stimuli. This chapter discusses the classification, distribution, and functional properties of individual TRP channel types that have been implicated in various nociceptive and/or painful conditions.

  9. The control of transmembrane helix transverse position in membranes by hydrophilic residues.

    Science.gov (United States)

    Krishnakumar, Shyam S; London, Erwin

    2007-12-14

    The ability of hydrophilic residues to shift the transverse position of transmembrane (TM) helices within bilayers was studied in model membrane vesicles. Transverse shifts were detected by fluorescence measurements of the membrane depth of a Trp residue at the center of a hydrophobic sequence. They were also estimated from the effective length of the TM-spanning sequence, derived from the stability of the TM configuration under conditions of negative hydrophobic mismatch. Hydrophilic residues (at the fifth position in a 21-residue hydrophobic sequence composed of alternating Leu and Ala residues and flanked on both ends by two Lys) induced transverse shifts that moved the hydrophilic residue closer to the membrane surface. At pH 7, the dependence of the extent of shift upon the identity of the hydrophilic residue increased in the order: L snorkeling). Additional experiments showed that shift was also modulated by the position of the hydrophilic residue in the sequence and the hydrophobicity of the sequence moved out of the bilayer core upon shifting. Combined, these studies show that the insertion boundaries of TM helices are very sensitive to sequence, and can be altered even by weakly hydrophilic residues. Thus, many TM helices may have the capacity to exist in more than one transverse position. Knowledge of the magnitudes of transverse shifts induced by different hydrophilic residues should be useful for design of mutagenesis studies measuring the effect of transverse TM helix position upon function.

  10. Label-free and multiplex detection of antibiotic residues in milk using imaging surface plasmon resonance-based immunosensor.

    Science.gov (United States)

    Rebe Raz, Sabina; Bremer, Maria G E G; Haasnoot, Willem; Norde, Willem

    2009-09-15

    Monitoring of antimicrobial drug residues in foods relies greatly on the availability of adequate analytical techniques. Currently, there is a need for a high-throughput screening method with a broad-spectrum detection range. This paper describes the development of a microarray biosensor, based on an imaging surface plasmon resonance (iSPR) platform, for quantitative and simultaneous immunodetection of different antibiotic residues in milk. Model compounds from four major antibiotic families: aminoglycosides (Neomycin, Gentamicin, Kanamycin, and Streptomycin), sulfonamides (Sulfamethazine), fenicols (Chloramphenicol), and fluoroquinolones (Enrofloxacin) were detected using a single sensor chip. By multiplexing seven immunoassays in a competitive format, we were able to measure all the target compounds at parts per billion (ppb) levels in buffer and in 10x-diluted milk. The assays for Neomycin, Kanamycin, Streptomycin, Enrofloxacin, and Sulfamethazine were sensitive enough for milk control at maximum residue levels as established in the European Union. The overall performance of the biosensor was determined to be comparable to that of conventional four-channel surface plasmon resonance (SPR)-based biosensors, in terms of assay sensitivity and robustness. Combining the advantages of a SPR sensor and a microarray, utilization of the biosensor described here offers a promising alternative to the existing methods and is highly relevant for multianalyte food profiling.

  11. Antimutagenic effect of resveratrol against Trp-P-1.

    Science.gov (United States)

    Uenobe, F; Nakamura, S; Miyazawa, M

    1997-02-03

    The methanol extract of Yucca schidigera (YE) showed a suppressive effect on umu gene expression of the SOS response induced by 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) in Salmonella typhimurium TA1535/pSK1002. The suppressive effect of YE was also observed for 2-aminoanthracene and activated Trp-P-1, without a significant effect on bacterial growth. The extract exhibited a weak suppressive effect on SOS-induction by N-methyl-N'-nitro-N-nitrosoguanidine, but not by furylfuramide or 4-nitroquinoline-1-oxide. The antimutagenic activity of YE against Trp-P-1 was demonstrated by Ames assay using Salmonella typhimurium TA98. Isolation and purification of the active component of YE was carried out using SiO2 column chromatography, and 275 mg of antimutagenic compound was isolated from 2.5 kg of dried chips of yucca roots and branches. The compound was identified as 3,4',5-trihydroxystilbene (THS). The SOS suppression and antimutagenicity of THS against Trp-P-1 was determined by umu test and Ames test.

  12. Transient receptor potential (TRP gene superfamily encoding cation channels

    Directory of Open Access Journals (Sweden)

    Pan Zan

    2011-01-01

    Full Text Available Abstract Transient receptor potential (TRP non-selective cation channels constitute a superfamily, which contains 28 different genes. In mammals, this superfamily is divided into six subfamilies based on differences in amino acid sequence homology between the different gene products. Proteins within a subfamily aggregate to form heteromeric or homomeric tetrameric configurations. These different groupings have very variable permeability ratios for calcium versus sodium ions. TRP expression is widely distributed in neuronal tissues, as well as a host of other tissues, including epithelial and endothelial cells. They are activated by environmental stresses that include tissue injury, changes in temperature, pH and osmolarity, as well as volatile chemicals, cytokines and plant compounds. Their activation induces, via intracellular calcium signalling, a host of responses, including stimulation of cell proliferation, migration, regulatory volume behaviour and the release of a host of cytokines. Their activation is greatly potentiated by phospholipase C (PLC activation mediated by coupled GTP-binding proteins and tyrosine receptors. In addition to their importance in maintaining tissue homeostasis, some of these responses may involve various underlying diseases. Given the wealth of literature describing the multiple roles of TRP in physiology in a very wide range of different mammalian tissues, this review limits itself to the literature describing the multiple roles of TRP channels in different ocular tissues. Accordingly, their importance to the corneal, trabecular meshwork, lens, ciliary muscle, retinal, microglial and retinal pigment epithelial physiology and pathology is reviewed.

  13. Examination of Some Pesticide Residues in Surface Water, Sediment and Fish Tissue of Elechi Creek, Niger Delta, Nigeria

    Directory of Open Access Journals (Sweden)

    F. Upadhi

    2012-11-01

    Full Text Available An investigation into the levels of selected pesticides in surface waters, sediment and fish (Mudskipper in the Elechi creek was carried out in two seasons and at three pre-determined sites. A higher concentration of 2, 4-diamine was observed in all stations and matrixes, while propoxur was least observed. Pesticide concentration in sediment is significantly different from that of water and fish tissue, which had the highest residual concentration. Concentration of pesticides ranged from 0.01 to 0.04 μg/L in water, 0.01 to 0.06 μg/gdw in sediment and 0.01 to 0.07 μg/gdw in fish, respectively. There was bioaccumulation of pesticides in fish samples. Observed residues are generally higher than stipulated limit of 0.01 μg/L by USEPA for pesticides of aquatic life, therefore, possess an ecological risk to the ecosystem and consequently human health.

  14. Surface residues dynamically organize water bridges to enhance electron transfer between proteins

    CERN Document Server

    de la Lande, Aurélien; Řezáč, Jan; Sanders, Barry C; Salahub, Dennis R; 10.1073/pnas.0914457107

    2010-01-01

    Cellular energy production depends on electron transfer (ET) between proteins. In this theoretical study, we investigate the impact of structural and conformational variations on the electronic coupling between the redox proteins methylamine dehydrogenase and amicyanin from Paracoccus denitrificans. We used molecular dynamics simulations to generate configurations over a duration of 40ns (sampled at 100fs intervals) in conjunction with an ET pathway analysis to estimate the ET coupling strength of each configuration. In the wild type complex, we find that the most frequently occurring molecular configurations afford superior electronic coupling due to the consistent presence of a water molecule hydrogen-bonded between the donor and acceptor sites. We attribute the persistence of this water bridge to a "molecular breakwater" composed of several hydrophobic residues surrounding the acceptor site. The breakwater supports the function of nearby solvent-organizing residues by limiting the exchange of water molecul...

  15. [Distribution Characteristics and Risk Assessment of Organochlorine residues in Surface Soil of Pearl River Delta Economic Zone].

    Science.gov (United States)

    Dou, Lei; Yang, Guo-yi

    2015-08-01

    The contents of 17 Organochlorine pesticides (OCPs) of 605 samples collected from the surface of agriculture land from Pearl River Delta Economic Zone were determined by gas chromatography with electron capture detector (GC-ECD). The residual characteristics, sources and ecological risk of OCPs were also analyzed. The results showed that the detection rate of OCPs was 97.85%. The mean value of residue level was 20.67 microg x kg(-1), with the highest value of 649.33 microg x kg(-1). The main contaminants included DDTs, HCHs, endosulfan sulfate and methoxychlor. Compared with the soils of other cities, the levels of HCHs and DDTs in the studied area were arranged from low to middle levels. The OCPs were obviously regionally distributed. High content areas were mainly distributed in the central area with dense population, intense industrial and agricultural activities. The residue levels in different types of lands were significantly different: the arable land > garden land > woodland. Especially, the residue level was the highest in the vegetable land. Source analysis indicated that the HCHs might come from the use of lindane. DDTs in soil mainly came from early residues, but the dicofol might be the important source in partial area. Comparing the contents of HCHs and DDTs with our National Standard (GB 15618-2008), the qualified rates of the first and second standard of HCHs were 97.5% and 100%, respectively, and the DDTs were 95.5% and 97.7%, respectively. According to the risk assessment, DDTs may still have some potential ecological impact on the studied area.

  16. Environmental impact of APC residues from municipal solid waste incineration: reuse assessment based on soil and surface water protection criteria.

    Science.gov (United States)

    Quina, Margarida J; Bordado, João C M; Quinta-Ferreira, Rosa M

    2011-01-01

    Waste management and environmental protection are mandatory requirements of modern society. In our study, air pollution control (APC) residues from municipal solid waste incinerators (MSWI) were considered as a mixture of fly ash and fine particulate solids collected in scrubbers and fabric filters. These are hazardous wastes and require treatment before landfill. Although there are a number of treatment options, it is highly recommended to find practical applications rather than just dump them in landfill sites. In general, for using a construction material, beyond technical specifications also soil and surface water criteria may be used to ensure environmental protection. The Dutch Building Materials Decree (BMD) is a valuable tool in this respect and it was used to investigate which properties do not meet the threshold criteria so that APC residues can be further used as secondary building material. To this end, some scenarios were evaluated by considering release of inorganic species from unmoulded and moulded applications. The main conclusion is that the high amount of soluble salts makes the APC residues a building material prohibited in any of the conditions tested. In case of moulding materials, the limits of heavy metals are complied, and their use in Category 1 would be allowed. However, also in this case, the soluble salts lead to the classification of "building material not allowed". The treatments with phosphates or silicates are able to solve the problem of heavy metals, but difficulties with the soluble salts are still observed. This analysis suggests that for APC residues to comply with soil and surface water protection criteria to be further used as building material at least a pre-treating for removing soluble salts is absolutely required.

  17. Effect of Plastic Pre-straining on Residual Stress and Composition Profiles in Low-Temperature Surface-Hardened Austenitic Stainless Steel

    DEFF Research Database (Denmark)

    Bottoli, Federico; Christiansen, Thomas Lundin; Winther, Grethe

    2016-01-01

    or nitrocarburized. The residual stress profiles resulting from the thermochemical low-temperature surface treatment were measured. The results indicate high-residual compressive stresses of several GPa’s in the nitrided region, while lower-compressive stresses are produced in the carburized case. Plastic...

  18. The influence of solder mask and hygroscopic flux residues on water layer formation on PCBA surface and corrosion reliability of electronics

    DEFF Research Database (Denmark)

    Piotrowska, Kamila; Jellesen, Morten Stendahl; Ambat, Rajan

    2017-01-01

    The presence of solder flux residue on the Printed Circuit Board Assembly (PCBA) surface compromises the corrosion reliability of electronics under humid conditions and can lead to degradation of the device’s lifetime. In this work, the effect of solder mask morphology and hygroscopic residues were...

  19. Evaluation of Residual Stress Relaxation in Surface-Treated Engine Alloys

    Science.gov (United States)

    2008-06-01

    account for the inevitable stress release during this process. This method is inherently destructive since it leaves a deep hole on the surface...accelerator or a nuclear reactor. 3 Surface enhancement methods, such as shot peening, laser shock peening, and low- plasticity burnishing , significantly...peening, laser shock peening, or low-plasticity burnishing , surface treatment also leaves substantial microstructural damage behind in the material

  20. Towards self-tuning residual generators for UAV control surface fault diagnosis

    DEFF Research Database (Denmark)

    Blanke, Mogens; Hansen, Søren

    2013-01-01

    Control surface fault diagnosis is essential for timely detection of manoeuvring and stability risks for an unmanned aircraft. Timely detection is crucial since control surface related faults impact stability of flight and safety. Reliable diagnosis require well fitting dynamical models but with ...... flights with different members of a population of UAVs that have inherent model uncertainty from one member to another and from one flight to another. Events with actual faults on control surfaces demonstrates the efficacy of the approach....

  1. Influence of sodium chloride and weak organic acids (flux residues) on electrochemical migration of tin on surface mount chip components

    DEFF Research Database (Denmark)

    Verdingovas, Vadimas; Jellesen, Morten Stendahl; Ambat, Rajan

    2013-01-01

    The electrolytic properties of sodium chloride and no-clean solder flux residue, and their effects on electrochemical migration and dendrite growth on surface mount chip capacitors were investigated. The leakage current dependency on concentration of contaminants was measured by a solution...... showed a difference which is caused by polarization effects, and demonstrated existing issues when indexing contamination levels on printed circuit board assemblies using a standardised solvent extract method. The experimental results showed that dendrite growth was dependent on the type and amount...

  2. Detection of fresh-cut produce processing residues on food contact surface materials using hyperspectral imaging

    Science.gov (United States)

    To reduce the risk of foodborne-illness, produce processors currently clean and sanitize food contact surfaces daily before production starts. Current methods to verify the efficacy of cleaning procedures include visual inspection and direct surface sampling using ATP bioluminescence assays and cul...

  3. File list: Oth.EmF.05.Trp53.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.EmF.05.Trp53.AllCell mm9 TFs and others Trp53 Embryonic fibroblast SRX483599,SR...X270554,SRX270556 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.EmF.05.Trp53.AllCell.bed ...

  4. File list: Oth.ALL.20.Trp53.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.20.Trp53.AllCell mm9 TFs and others Trp53 All cell types SRX483599,SRX27055...4,SRX335560,SRX270556 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.ALL.20.Trp53.AllCell.bed ...

  5. File list: Oth.EmF.20.Trp53.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.EmF.20.Trp53.AllCell mm9 TFs and others Trp53 Embryonic fibroblast SRX483599,SR...X270554,SRX270556 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.EmF.20.Trp53.AllCell.bed ...

  6. File list: Oth.ALL.05.Trp53.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.05.Trp53.AllCell mm9 TFs and others Trp53 All cell types SRX483599,SRX27055...4,SRX335560,SRX270556 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.ALL.05.Trp53.AllCell.bed ...

  7. File list: Oth.EmF.10.Trp53.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.EmF.10.Trp53.AllCell mm9 TFs and others Trp53 Embryonic fibroblast SRX483599,SR...X270554,SRX270556 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.EmF.10.Trp53.AllCell.bed ...

  8. File list: Oth.ALL.10.Trp53.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.10.Trp53.AllCell mm9 TFs and others Trp53 All cell types SRX483599,SRX27055...4,SRX335560,SRX270556 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.ALL.10.Trp53.AllCell.bed ...

  9. File list: Oth.ALL.50.Trp53.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.50.Trp53.AllCell mm9 TFs and others Trp53 All cell types SRX483599,SRX27055...4,SRX335560,SRX270556 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.ALL.50.Trp53.AllCell.bed ...

  10. Monitoring the Folding of Trp-cage Peptide by Two-dimensional Infrared (2DIR) Spectroscopy

    Science.gov (United States)

    Lai, Zaizhi; Preketes, Nicholas K.; Mukamel, Shaul; Wang, Jin

    2013-01-01

    Protein folding is one of the most fundamental problems in modern molecular biology. Uncovering the detailed folding mechanism requires methods that can monitor the structures at high temporal and spatial resolution. Two-dimensional infrared (2DIR) spectroscopy is a new tool for studying protein structures and dynamics with high time resolution. Using atomistic molecular dynamics simulations, we illustrate the folding process of Trp-cage along the dominant pathway on free energy landscape by analyzing nonchiral and chiral coherent 2DIR spectra along the pathway. Isotope-labeling is used to reveal residue-specific information. We show that the high resolution structural sensitivity of 2DIR can differentiate the ensemble evolution of protein, and thus provides a microscopic picture of the folding process. PMID:23448437

  11. Nutrient removal capacity of wood residues for the Agro-environmental safety of ground and surface waters

    Directory of Open Access Journals (Sweden)

    Paulo A. Dumont

    2014-07-01

    Full Text Available The aim of this study was to determine the effectiveness of wood residues in the removal of nutrients (ammonium-N; NH4-N from nutrient-rich (NH4-N waters. The water holding capacity of the wood materials was also determined. Carried out at Rothamsted Research, North Wyke, UK, this controlled laboratory experiment tested two wood residues; in length, one being 1-2cm and the other from 150 µm (microns to 9.5mm. Although a wide range of studies have shown the effectiveness and performance of various absorbent materials as animal beddings, such as straw (cereal straw, woodchip (sawdust, bark or wood shavings, bracken and rushes, only few have focused on the NH4-N sorption/desorption capacity. The depuration capacity of wood residues from nutrient-rich effluents such as those from cattle bedded on woodchip or straw will be controlled by processes such as sorption (adsorption-absorption and desorption of nutrients. Studies have reported the nitrogen removal capacity of woodchip materials and biochar from woodchip as well as removal of NH4+-N from domestic and municipal wastewater, farm dirty water, landfill and industry effluents. These studies have observed that the mechanism of removal of nitrogen is by either increasing NO3--N removal form leachate by enhancing N2O losses via denitrification (biochar as carbon source for denitrifiers or by decreasing NH4+-N in leachate through adsorption to negatively charged sites. Results showed that although the cation exchange capacity (CEC and surface area (SA are both fundamental properties of adsorbent materials, no correlation was found with CEC and adsorption or desorption. Nor did changes in pH appear to be sufficiently important to cause changes in CEC. For this reason, osmotic pressure appeared to be a more predominant parameter controlling processes of adsorption and desorption of NH4+-N in both wood residues. Thus, wood residues high in NH4+-N should be avoided, as they could have an opposite effect

  12. Detection of poly(ethylene glycol) residues from nonionic surfactants in surface water by1h and13c nuclear magnetic resonance spectrometry

    Science.gov (United States)

    Leenheer, J.A.; Wershaw, R. L.; Brown, P.A.; Noyes, T.I.

    1991-01-01

    ??? Poly(ethylene glycol) (PEG) residues were detected in organic solute isolates from surface water by 1H nuclear magnetic resonance spectrometry (NMR), 13C NMR spectrometry, and colorimetric assay. PEG residues were separated from natural organic solutes in Clear Creek, CO, by a combination of methylation and chromatographic procedures. The isolated PEG residues, characterized by NMR spectrometry, were found to consist of neutral and acidic residues that also contained poly(propylene glycol) moieties. The 1H NMR and the colorimetric assays for poly(ethylene glycol) residues were done on samples collected in the lower Mississippi River and tributaries between St. Louis, MO, and New Orleans, LA, in July-August and November-December 1987. Aqueous concentrations for poly(ethylene glycol) residues based on colorimetric assay ranged from undetectable to ???28 ??g/L. Concentrations based on 1H NMR spectrometry ranged from undetectable to 145 ??g/L.

  13. Residual viral and bacterial contamination of surfaces after cleaning and disinfection

    NARCIS (Netherlands)

    Tuladhar, E.; Hazeleger, W.C.; Koopmans, M.; Zwietering, M.H.; Beumer, R.R.; Duizer, E.

    2012-01-01

    Environmental surfaces contaminated with pathogens can be sources of indirect transmission, and cleaning and disinfection are common interventions focused on reducing contamination levels. We determined the efficacy of cleaning and disinfection procedures for reducing contamination by noroviruses, r

  14. Residual viral and bacterial contamination of surfaces after cleaning and disinfection

    NARCIS (Netherlands)

    Tuladhar, E.; Hazeleger, W.C.; Koopmans, M.; Zwietering, M.H.; Beumer, R.R.; Duizer, E.

    2012-01-01

    Environmental surfaces contaminated with pathogens can be sources of indirect transmission, and cleaning and disinfection are common interventions focused on reducing contamination levels. We determined the efficacy of cleaning and disinfection procedures for reducing contamination by noroviruses,

  15. Gold-plating of Mylar lift films to capitalize on surface enhanced Raman spectroscopy for chemical extraction of drug residues.

    Science.gov (United States)

    Fox, James D; Waverka, Kristin N; Verbeck, Guido F

    2012-03-10

    The method of residue extraction through electrostatic lifting provides a distinctive mode of performing ultra-trace analysis. These lifts provide a medium for analyte extraction via nanomanipulation-coupled to nanospray ionization-mass spectrometry (NSI-MS). This method of extraction can be coupled to Raman spectroscopy for supplemental verification of analytes using surface enhanced Raman scattering (SERS). The gold surface used for SERS provides an enhanced effect on peak signal intensity allowing ultra-trace amounts to be detected more effectively. The aim of this research is to utilize gold-coated films with electrostatic lifting in order to collect latent materials and analyze chemicals of interest contained in them via SERS.

  16. Development and validation of a high-performance liquid chromatographic method for the determination of clomazone residues in surface water.

    Science.gov (United States)

    Zanella, R; Primel, E G; Gonçalves, F F; Martins, A F

    2000-12-29

    A method is described for the determination of clomazone residues in surface water by high-performance liquid chromatography with UV detection. The method involves solid-phase extraction with C18 extraction tubes. Clomazone was separated on a C18 column with a mobile phase of methanol-water (65:35, v/v) at pH 4.0 and a flow-rate of 1.0 ml/min. After optimization of the extraction and separation conditions, the method was validated. The method developed can be used for determination of clomazone in surface water, at the limit of 0.1 mcirog/l set by the European Union drinking water directive, with a 400-fold preconcentration.

  17. Effect of Interaction of Non-residual Fractions on Adsorption of Atrazine onto Surficial Sediments and Natural Surface Coating Samples

    Institute of Scientific and Technical Information of China (English)

    LI Yu; LI Shan-shan; GAO Qian; WANG Ao

    2011-01-01

    To quantify the effect of the interaction of non-residual fractions[Fe oxides(Fe), Mn oxide(Mn), organic materials(OMs)] in the surficial sediments and the natural surface coating samples on the adsorption of atrazine(AT),an AT multiple regression adsorption modeI(AT-MRAM) was developed. The AT-MRAM improves upon the previous AT additional adsorption modeI(AT-AAM) with superior goodness-of-fit test(adjusted R2=ca.l.000), F-test and t-test(P<0.01), and reveals the effect of the interaction among the components in the surficial sediments(SSs) and natural surface coatings samples(NSCSs) on the adsorption of AT, which was neglected by the AT-AAM. Meanwhile, the AT-MRAM was also verified through adsorption experiments of AT and the relative deviation between predicted maximum adsorption of AT and the experimental one is less than 15%. The resulted information shows that Mn is prone to interact with other non-residual components, the total maximum adsorption of AT is inversly proportional to the level of Mn, and Fe and OMs facilitate the adsorption of AT. The results also indicate that the adsorption of AT is not only dominated by Fe, OMs, Fe/OMs, but also restrained by Fe/Mn, Fe/Mn/OMs, with lesser roles attributed to Mn, and the estimated AT distributions among the components do not agree with that previously predicted by the AT-AAM, especially with the relative contribution of Mn to the adsorption of AT, revealing significant contribution of the interactions among non-residual components in controlling the behavior of AT in aquatic environments.

  18. Drosophila sperm surface alpha-L-fucosidase interacts with the egg coats through its core fucose residues.

    Science.gov (United States)

    Intra, Jari; Concetta, Veltri; Daniela, De Caro; Perotti, Maria Elisa; Pasini, Maria Enrica

    2015-08-01

    Sperm-oocyte interaction during fertilization is multiphasic, with multicomponent events, taking place between egg's glycoproteins and sperm surface receptors. Protein-carbohydrate complementarities in gamete recognition have observed in cases throughout the whole evolutionary scale. Sperm-associated α-L-fucosidases have been identified in various organisms. Their wide distribution and known properties reflect the hypothesis that fucose and α-L-fucosidases have fundamental function(s) during gamete interactions. An α-L-fucosidase has been detected as transmembrane protein on the surface of spermatozoa of eleven species across the genus Drosophila. Immunofluorescence labeling showed that the protein is localized in the sperm plasma membrane over the acrosome and the tail, in Drosophila melanogaster. In the present study, efforts were made to analyze with solid phase assays the oligosaccharide recognition ability of fruit fly sperm α-L-fucosidase with defined carbohydrate chains that can functionally mimic egg glycoconjugates. Our results showed that α-L-fucosidase bound to fucose residue and in particular it prefers N-glycans carrying core α1,6-linked fucose and core α1,3-linked fucose in N-glycans carrying only a terminal mannose residue. The ability of sperm α-L-fucosidase to bind to the micropylar chorion and to the vitelline envelope was examined in in vitro assays in presence of α-L-fucosidase, either alone or in combination with molecules containing fucose residues. No binding was detected when α-L-fucosidase was pre-incubated with fucoidan, a polymer of α-L-fucose and the monosaccharide fucose. Furthermore, egg labeling with anti-horseradish peroxidase, that recognized only core α1,3-linked fucose, correlates with α-L-fucosidase micropylar binding. Collectively, these data support the hypothesis of the potential role of this glycosidase in sperm-egg interactions in Drosophila.

  19. The surface chemistry resulting from low-pressure plasma treatment of polystyrene: The effect of residual vessel bound oxygen

    Science.gov (United States)

    Dhayal, Marshal; Alexander, Morgan R.; Bradley, James W.

    2006-09-01

    The surface chemistry of plasma treated polystyrene samples has been studied in a specially designed low-pressure argon discharge system incorporating in situ XPS analysis. By using an electrostatic grid biasing technique, the plasma source can also be used in a mode preventing ion interactions with the sample. The system, which utilizes a vacuum transfer chamber between plasma and XPS analysis has allowed us to differentiate between the level of oxygen incorporated at the polystyrene surface from residual gas during treatment and that from the exposure of the treated sample to the laboratory atmosphere. Using typical base pressures of about 5 × 10 -3 Pa (4 × 10 -5 Torr) the XPS results show that significant oxygen surface incorporation resulted from oxygen containing species in the plasma itself (i.e. water vapour with 2 × 10 -3 Pa partial pressure). The surface concentration of O was measured at 7.6 at.%. Subsequent atmospheric exposure of the treated samples resulted in only a small increase (of 0.6 at.%) in oxygen incorporation in the form of acid anhydride functionalities. XPS measurements of PS samples exposed to plasmas with no ion-surface component (i.e. exposure from VUV, UV and excited neutral species only) showed no appreciable change in oxygen incorporation compared to those with low-energy ion bombardment from the plasma (free radical sites in this discharge regime.

  20. Effect of Basic Residue on the Kinetics of Peptide Fragmentation Examined Using Surface-Induced Dissociation Combined with Resonant Ejection

    Energy Technology Data Exchange (ETDEWEB)

    Laskin, Julia

    2015-11-30

    In this work, resonant ejection coupled with surface-induced dissociation (SID) in a Fourier transform ion cyclotron resonance mass spectrometer is used to examine fragmentation kinetics of two singly protonated hexapeptides, RYGGFL and KYGGFL, containing the basic arginine residue and less basic lysine residue at the N-terminus. The kinetics of individual reaction channels at different collision energies are probed by applying a short ejection pulse (1 ms) in resonance with the cyclotron frequency of a selected fragment ion and varying the delay time between ion-surface collision and resonant ejection while keeping total reaction delay time constant. Rice-Ramsperger-Kassel-Marcus (RRKM) modeling of the experimental data provides accurate threshold energies and activation entropies of individual reaction channels. Substitution of arginine with less basic lysine has a pronounced effect on the observed fragmentation kinetics of several pathways, including the b2 ion formation, but has little or no effect on formation of the b5+H2O fragment ion. The combination of resonant ejection SID, time- and collision energy-resolved SID, and RRKM modeling of both types of experimental data provides a detailed mechanistic understanding of the primary dissociation pathways of complex gaseous ions.

  1. Trypanosoma evansi: identification and characterization of a variant surface glycoprotein lacking cysteine residues in its C-terminal domain.

    Science.gov (United States)

    Jia, Yonggen; Zhao, Xinxin; Zou, Jingru; Suo, Xun

    2011-01-01

    African trypanosomes are flagellated unicellular parasites which proliferate extracellularly in the mammalian host blood-stream and tissue spaces. They evade the hosts' antibody-mediated lyses by sequentially changing their variant surface glycoprotein (VSG). VSG tightly coats the entire parasite body, serving as a physical barrier. In Trypanosoma brucei and the closely related species Trypanosoma evansi, Trypanosoma equiperdum, each VSG polypeptide can be divided into N- and C-terminal domains, based on cysteine distribution and sequence homology. N-terminal domain, the basis of antigenic variation, is hypervariable and contains all the exposed epitopes; C-terminal domain is relatively conserved and a full set of four or eight cysteines were generally observed. We cloned two genes from two distinct variants of T. evansi, utilizing RT-PCR with VSG-specific primers. One contained a VSG type A N-terminal domain followed a C-terminal domain lacking cysteine residues. To confirm that this gene is expressed as a functional VSG, the expression and localization of the corresponding gene product were characterized using Western blotting and immunofluorescent staining of living trypanosomes. Expression analysis showed that this protein was highly expressed, variant-specific, and had a ubiquitous cellular surface localization. All these results indicated that it was expressed as a functional VSG. Our finding showed that cysteine residues in VSG C-terminal domain were not essential; the conserved C-terminal domain generally in T. brucei like VSGs would possibly evolve for regulating the VSG expression.

  2. New insights into TRP channels: Interaction with pattern recognition receptors.

    Science.gov (United States)

    Han, Huirong; Yi, Fan

    2014-01-01

    An increasing number of studies have implicated that the activation of innate immune system and inflammatory mechanisms are of importance in the pathogenesis of numerous diseases. The innate immune system is present in almost all multicellular organisms in response to pathogens or tissue injury, which is performed via germ-line encoded pattern-recognition receptors (PRRs) to recognize pathogen-associated molecular patterns (PAMPs) or dangers-associated molecular patterns (DAMPs). Intracellular pathways linking immune and inflammatory response to ion channel expression and function have been recently identified. Among ion channels, transient receptor potential (TRP) channels are a major family of non-selective cation-permeable channels that function as polymodal cellular sensors involved in many physiological and pathological processes. In this review, we summarize current knowledge about classifications, functions, and interactions of TRP channels and PRRs, which may provide new insights into their roles in the pathogenesis of inflammatory diseases.

  3. Agro-Residues: Surface Treatment and Characterization of Date Palm Tree Fiber as Composite Reinforcement

    Directory of Open Access Journals (Sweden)

    Elsayed A. Elbadry

    2014-01-01

    Full Text Available The aims of this research are to investigate the effect of different surface treatment methods on the different properties of date palm fiber (DPF compared to raw DPF fibers such as surface morphology, density, thermal stability, and tensile properties. The first surface treatment is called surface hand cleaning which can be carried out by cleaning the fibers by soft sand cloth; the second one is the same as the first one after DPF heat treatment in the furnace at 100°C for 1.5 h and the third one is by chemical treatment with 1% NaOH at 100°C for 1 h. The results showed that the mechanical performance of DPF was enhanced by the different treatments and the chemical treatment has pronounced effect on the behavior of DPF. Raw fibers showed the highest variability and presented the lowest value of Weibull modulus, whereas the fibers showed less variability by carrying out the different treatments. Moreover, using soda treatment cleans the fiber surface which causes fibrillation and therefore the tensile strength of the fibers increases.

  4. TRP channels in prostate cancer: the good,the bad and the ugly?

    Institute of Scientific and Technical Information of China (English)

    Dimitra Gkikal; Natalia Prevarskaya

    2011-01-01

    @@ During the last decade,transient receptor potential (TRP) channels emerge as key proteins in central mechanisms of the carcinogenesis such as cell proliferation,apoptosis and migration.Initial studies showed that expression profile of some TRP channels,notably TRP melastatin 8 (TRPM8),TRP vanilloid 6 (TRPV6),TRP canonical (TRPC6) and TRPV2,is changing during the development and the progression of prostate cancer towards the hormone-refractory stages.The link between the change in expression levels and the functional role of these channels in prostate cancer is step by step being elucidated.These recent advances are here described and discussed.

  5. Studies on the biotin-binding site of avidin. Tryptophan residues involved in the active site.

    OpenAIRE

    Gitlin, G; Bayer, E A; Wilchek, M

    1988-01-01

    Egg-white avidin was modified with the tryptophan-specific reagent 2-hydroxy-5-nitrobenzyl bromide. The complete loss of biotin-binding activity was achieved upon modification of an average of one tryptophan residue per avidin subunit. The identity of the modified residues was determined by isolating the relevant tryptic and chymotryptic peptides from CNBr-cleaved avidin fragments. The results demonstrate that Trp-70 and Trp-110 are modified in approximately equivalent proportions. It is beli...

  6. Aire deficiency promotes TRP-1 specific immune rejection of melanoma

    Science.gov (United States)

    Zhu, Meng-Lei; Nagavalli, Anil; Su, Maureen A.

    2013-01-01

    The thymic transcription factor AIRE prevents autoimmunity in part by promoting expression of tissue-specific self-antigens, which include many cancer antigens. For example, AIRE-deficient patients are predisposed to vitiligo, an autoimmune disease of melanocytes that is often triggered by efficacious immunotherapies against melanoma. Therefore, we hypothesized that Aire deficiency in mice may elevate immune responses to cancer and provide insights into how such responses might be triggered. In this study, we show that Aire deficiency decreases thymic expression of TRP-1 (TYRP1), which is a self-antigen in melanocytes and a cancer antigen in melanomas. Aire deficiency resulted in defective negative selection of TRP-1 specific T cells without affecting thymic numbers of regulatory T cells. Aire deficient mice displayed elevated T cell immune responses that were associated with suppression of melanoma outgrowth. Further, transplantation of Aire-deficient thymic stroma was sufficient to confer more effective immune rejection of melanoma in an otherwise Aire wildtype host. Together, our work showed how Aire deficiency can enhance immune responses against melanoma, and how manipulating TRP-1 specific T cell negative selection may offer a logical strategy to enhance immune rejection of melanoma. PMID:23370329

  7. Aire deficiency promotes TRP-1-specific immune rejection of melanoma.

    Science.gov (United States)

    Zhu, Meng-Lei; Nagavalli, Anil; Su, Maureen A

    2013-04-01

    The thymic transcription factor autoimmune regulator (Aire) prevents autoimmunity in part by promoting expression of tissue-specific self-antigens, which include many cancer antigens. For example, AIRE-deficient patients are predisposed to vitiligo, an autoimmune disease of melanocytes that is often triggered by efficacious immunotherapies against melanoma. Therefore, we hypothesized that Aire deficiency in mice may elevate immune responses to cancer and provide insights into how such responses might be triggered. In this study, we show that Aire deficiency decreases thymic expression of TRP-1 (TYRP1), which is a self-antigen in melanocytes and a cancer antigen in melanomas. Aire deficiency resulted in defective negative selection of TRP-1-specific T cells without affecting thymic numbers of regulatory T cells. Aire-deficient mice displayed elevated T-cell immune responses that were associated with suppression of melanoma outgrowth. Furthermore, transplantation of Aire-deficient thymic stroma was sufficient to confer more effective immune rejection of melanoma in an otherwise Aire wild-type host. Together, our work showed how Aire deficiency can enhance immune responses against melanoma and how manipulating TRP-1-specific T-cell negative selection may offer a logical strategy to enhance immune rejection of melanoma. ©2013 AACR.

  8. TRP channel gene expression in the mouse retina.

    Science.gov (United States)

    Gilliam, Jared C; Wensel, Theodore G

    2011-12-08

    In order to identify candidate cation channels important for retinal physiology, 28 TRP channel genes were surveyed for expression in the mouse retina. Transcripts for all TRP channels were detected by RT-PCR and sequencing. Northern blotting revealed that mRNAs for 12 TRP channel genes are enriched in the retina. The strongest signals were observed for TRPC1, TRPC3, TRPM1, TRPM3, and TRPML1, and clear signals were obtained for TRPC4, TRPM7, TRPP2, TRPV2, and TRPV4. In situ hybridization and immunofluorescence revealed widespread expression throughout multiple retinal layers for TRPC1, TRPC3, TRPC4, TRPML1, PKD1, and TRPP2. Striking localization of enhanced mRNA expression was observed for TRPC1 in the photoreceptor inner segment layer, for TRPM1 in the inner nuclear layer (INL), for TRPM3 in the INL, and for TRPML1 in the outer plexiform and nuclear layers. Strong immunofluorescence signal in cone outer segments was observed for TRPM7 and TRPP2. TRPC5 immunostaining was largely confined to INL cells immediately adjacent to the inner plexiform layer. TRPV2 antibodies stained photoreceptor axons in the outer plexiform layer. Expression of TRPM1 splice variants was strong in the ciliary body, whereas TRPM3 was strongly expressed in the retinal pigmented epithelium.

  9. The assessment of ultrasonic root surface debridement by determination of residual endotoxin levels.

    Science.gov (United States)

    Smart, G J; Wilson, M; Davies, E H; Kieser, J B

    1990-03-01

    Recent demonstrations of the superficial nature and ease of removal of root-surface-associated cytotoxic materials suggest that extensive root-surface instrumentation is not warranted clinically. This in vitro investigation determined the detoxifying effects of a conservative regime of ultrasonic root debridement, using the Limulus amoebocyte lysate assay for lipopolysaccharide (LPS) as an indication of the presence or absence of cytotoxic materials. 20 extracted periodontally-involved single-rooted teeth, with no clinically detectable calculus deposits, were debrided with a Cavitron TF-10 tip. Light pressure, resulting from a force application of approximately 50 g, lasting for 0.8 s/mm2 of root surface was used and complete overlapping instrumentation ensured. The finding of LPS levels of less than 2.5 ng per root in 19 out of 20 teeth after such debridement was comparable to LPS levels found on healthy, uninvolved control teeth, and thus endorses the growing belief that root surface cleanliness can be readily achieved.

  10. Quantifying the size-dependent effect of the residual surface stress on the resonant frequencies of silicon nanowires if finite deformation kinematics are considered.

    Science.gov (United States)

    Park, Harold S

    2009-03-18

    There are two major objectives to the present work. The first objective is to demonstrate that, in contrast to predictions from linear surface elastic theory, when nonlinear, finite deformation kinematics are considered, the residual surface stress does impact the resonant frequencies of silicon nanowires. The second objective of this work is to delineate, as a function of nanowire size, the relative contributions of both the residual (strain-independent) and the surface elastic (strain-dependent) parts of the surface stress to the nanowire resonant frequencies. Both goals are accomplished by using the recently developed surface Cauchy-Born model, which accounts for nanoscale surface stresses through a nonlinear, finite deformation continuum mechanics model that leads to the solution of a standard finite element eigenvalue problem for the nanowire resonant frequencies. In addition to demonstrating that the residual surface stress does impact the resonant frequencies of silicon nanowires, we further show that there is a strong size dependence to its effect; in particular, we find that consideration of the residual surface stress alone leads to significant errors in predictions of the nanowire resonant frequency, with an increase in error with decreasing nanowire size. Correspondingly, the strain-dependent part of the surface stress is found to have an increasingly important effect on the resonant frequencies of the nanowires with decreasing nanowire size.

  11. Optimization of low-cost biosurfactant production from agricultural residues through response surface methodology.

    Science.gov (United States)

    Ebadipour, N; Lotfabad, T Bagheri; Yaghmaei, S; RoostaAzad, R

    2016-01-01

    Biosurfactants are surface-active compounds capable of reducing surface tension and interfacial tension. Biosurfactants are produced by various microorganisms. They are promising replacements for chemical surfactants because of biodegradability, nontoxicity, and their ability to be produced from renewable sources. However, a major obstacle in producing biosurfactants at the industrial level is the lack of cost-effectiveness. In the present study, by using corn steep liquor (CSL) as a low-cost agricultural waste, not only is the production cost reduced but a higher production yield is also achieved. Moreover, a response surface methodology (RSM) approach through the Box-Behnken method was applied to optimize the biosurfactant production level. The results found that biosurfactant production was improved around 2.3 times at optimum condition when the CSL was at a concentration of 1.88 mL/L and yeast extract was reduced to 25 times less than what was used in a basic soybean oil medium (SOM). The predicted and experimental values of responses were in reasonable agreement with each other (Pred-R(2) = 0.86 and adj-R(2) = 0.94). Optimization led to a drop in raw material price per unit of biosurfactant from $47 to $12/kg. Moreover, the biosurfactant product at a concentration of 84 mg/L could lower the surface tension of twice-distilled water from 72 mN/m to less than 28 mN/m and emulsify an equal volume of kerosene by an emulsification index of (E24) 68% in a two-phase mixture. These capabilities made these biosurfactants applicable in microbial enhanced oil recovery (MEOR), hydrocarbon remediation, and all other petroleum industry surfactant applications.

  12. Eddy Current Residual Stress Profiling in Surface-Treated Engine Alloys

    Science.gov (United States)

    2009-06-01

    during this process. This method is inherently destructive since it leaves a deep hole on the surface. Although the accuracy of XRD measurements is quite...SP), laser shock peening (LSP), and low-plasticity burnishing (LPB), significantly improve the fatigue resistance and foreign object damage...and XRD results in low-plasticity burnished Waspaloy of approximately 15% maximum plastic strain. There is a fairly good agreement between the non

  13. Requiring Pollutant Discharge Permits for Pesticide Applications that Deposit Residues in Surface Waters

    OpenAIRE

    Terence Centner; Nicholas Eberhart

    2014-01-01

    Agricultural producers and public health authorities apply pesticides to control pests that damage crops and carry diseases. Due to the toxic nature of most pesticides, they are regulated by governments. Regulatory provisions require pesticides to be registered and restrictions operate to safeguard human health and the environment. Yet pesticides used near surface waters pose dangers to non-target species and drinking water supplies leading some governments to regulate discharges of pesticid...

  14. Mutagenic analysis of putative domain II and surface residues in mosquitocidal Bacillus thuringiensis Cry19Aa toxin.

    Science.gov (United States)

    Roh, Jong Yul; Nair, Manoj S; Liu, Xinyan Sylvia; Dean, Donald H

    2009-06-01

    The mosquitocidal crystal protein, Cry19Aa, from Bacillus thuringiensis ssp. jegathesan, has high toxicity to Anopheles stephensi and Culex pipiens but is less toxic to Aedes aegypti. To study the functional role of putative domain II and surface residues in mosquito toxicity, 16 alanine substitution mutations were introduced into Cry19Aa. All mutant constructs were expressed as 65-kDa protoxins and subsequently digested by trypsin to produce further fragmented polypeptides of 40 and 25 kDa. With chymotrypsin, however, most protoxins were digested to 60 kDa and minor bands. The circular dichroism spectra of the chymotrypsin-activated toxins of Cry19Aa and muteins, Y324A, W357A, Y412A, Y414A, W416A, D418A and F485A indicated that there was no significant variation in their structure. In mosquito bioassays, Y324A, W357A, Y410A, W416A, D418A and F485A muteins showed substantial reductions in mosquitocidal activity toward A. aegypti and C. pipiens. These muteins also showed reduced competition with wild-type fluorescein 5-isothiocyanate-labeled Cry19Aa for binding to C. pipiens brush border membrane vesicles. These data suggest that the reduction of toxicity was a result of the reduced binding affinity. From these studies we have identified loop residues of domain II that are important in toxicity and receptor binding to Culex larval midgut.

  15. Structures of Cytochrome b 5 Mutated at the Charged Surface-Residues and Their Interactions with Cytochrome c

    Institute of Scientific and Technical Information of China (English)

    WU,Jian(邬键); WANG,Yun-Hua(王韵华); GAN,Jian-Hua(甘建华); WANG,Wen-Hu(王文虎); SUN,Bing-Yun(孙炳耘); HUANG,Zhong-Xian(黄仲贤); XIA,Zong-Xiang(夏宗芗)

    2002-01-01

    Glu44, Glu48, Glu56 and Asp60 are the negatively charged residues located at the molecular surface of cytochrome b5@Two mutants of cytochrome b5 were prepared, in which two or all of these four residues were mutated to alanines. The mutations give rise to slightly positive shifts of the redox potentials of cytochrome b5 and obvious decrease of the cytochrome b5-cytochrome c binding constants and electron transfer rates. The crystal structures of the two mutants were determined at 0.18 nm resolution, showing no alteration in overall structures and exhibiting slight chages in the local conformations around the mutation sites as compared with the wild-type protein. Based on the crystal structure of the quadruple-site mutant, a model for the binding of this mutant with cytochrome c is proposed, which involves the salt bridges from Glu37, Glu38 and heme propionate of cytochrome b5 to three lysines of cytochrome c and can well account for the properties and behaviors of this mutant.

  16. Radar response to crop residue cover and tillage application on postharvest agricultural surfaces

    Science.gov (United States)

    McNairn, Heather

    Les informations sur les pratiques de conservation des sols comme le labourage et la gestion des residus de culture sont requises afin d'estimer avec exactitude les risques d'erosion des sols. Quoique les micro-ondes soient sensibles aux conditions d'humidite et aux proprietes geometriques des surfaces, il n'en demeure pas moins que l'on connait encore peu sur la sensibilite des micro-ondes polarisees lineaires ou des parametres polarimetriques du ROS en fonction des caracteristiques des residus. A partir de donnees prises a l'aide d'un diffusometre monte sur un camion en 1996 et lors d'une mission SIR-C menee en 1994, cette recherche a demontre que les micro-ondes sont sensibles a la fois a la quantite et au type de couverture de residus, de meme qu'a la teneur en eau des residus. La reponse des polarisations croisees lineaires et de plusieurs parametres polarimetriques, incluant la hauteur pedestre, a permis d'observer qu'une diffusion volumique importante avait lieu en presence de vegetation senescente qui se tenait debout et pour les champs non laboures. La diffusion de surface dominait cependant pour les champs avec de faibles quantites de residus et des residus plus fins. La recherche a toutefois demontre que des conditions de surface complexes etaient crees par differentes combinaisons de residus et de pratiques de labourage. Par consequent, il faudra attendre que des donnees multi-polarisees ou polarimetriques soient acquises par les capteurs prevus a bord du satellite canadien RADARSAT-2 et du satellite ENVISAT de l'Agence spatiale europeenne avant de pouvoir completement caracteriser les champs apres la recolte.

  17. Preparation and characteri-zation of some surface nega-tively charged residue mu-tants of cytochrome b5

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Site-directed mutagenesis was used to obtain seven variants of tryptic fragment of bovine liver cytochrome b5 (cyt b5), in which the negatively charged residues around the heme exposed edge of cyt b5 were replaced by hydrophobic amino acid alanine. Double-site mutants, triple-site mutants and even quadruple-site mutants were obtained. DNA sequencing and molecular weight measurements of the mutant proteins both confirmed that these site-directed muta- genesises were successfully performed. Spectroelectrochemistry of these mutant proteins revealed that the apparent redox potentials of these mutant proteins caused a positive shift of 2-10 mV. The global structure of these mutant proteins did not show much difference from that of the wild type cyt b5, providing a solid base for the further study on the roles of the proteins' surface charges.

  18. Estimate of Small Stiffness and Damping Ratio in Residual Soil Using Spectral Analysis of Surface Wave Method

    Directory of Open Access Journals (Sweden)

    Bawadi Nor Faizah

    2016-01-01

    Full Text Available Research in the important parameters for modeling the dynamic behavior of soils has led to rapid development of the small strain stiffness and damping ratio for use in the seismic method. It is because, the experimental determination of the damping ratio is problematic, especially for hard soils sample. Many researchers have proved that the surface wave method is a reliable tool to determine shear wave velocity and damping ratio profiles at a site with very small strains level. Surface wave methods based on Rayleigh waves propagation and the resulting attenuation curve can become erroneous when higher modes contribute to the soil’s response. In this study, two approaches has been used to determine the shear strain amplitude and damping ratio of residual soils at small strain level using Spectral Analysis of Surface Wave (SASW method. One is to derive shear strain amplitude from the frequency-response curve and the other is to derive damping ratio from travel-time data. Then, the results are compared to the conventional method.

  19. Interrogation of the microstructure and residual stress of a nickel-base alloy subjected to surface severe plastic deformation

    Energy Technology Data Exchange (ETDEWEB)

    Ortiz, A.L. [Departamento de Ingenieria Mecanica, Energetica y de los Materiales, Universidad de Extremadura, 06071 Badajoz (Spain); Tian, J.W. [Department of Materials Science and Engineering, University of Tennessee, Knoxville, TN (United States); Villegas, J.C. [Intel Corporation, Chandler, AZ (United States); Shaw, L.L. [Department of Chemical, Materials and Biomolecular Engineering, University of Connecticut, Storrs, CT (United States)], E-mail: Leon.Shaw@Uconn.Edu; Liaw, P.K. [Department of Materials Science and Engineering, University of Tennessee, Knoxville, TN (United States)

    2008-02-15

    A low stacking-fault energy nickel-base, single-phase, face-centered-cubic (fcc) alloy has been subjected to surface severe plastic deformation (S{sup 2}PD) to introduce nano-grains and grain size gradients to the surface region of the alloy. The simultaneous microstructural and stress state changes induced by S{sup 2}PD have been investigated via the X-ray diffraction (XRD) analysis that includes evaluation of annealing and deformation twins, deformation faults, in-plane lattice parameters and elastic strains of the crystal lattice, macroscopic residual in-plane stresses, crystallite sizes, internal strains, dislocation densities, and crystallographic texture as a function of the depth measured from the processed surface. Microstructural changes have also been characterized using optical and electron microscopy in order to corroborate the findings from the XRD analysis. The results from the XRD analysis are in excellent agreement with those derived from the microscopy analysis. This is the first systematic and comprehensive study using XRD to quantify depth-profile changes in a wide range of microstructural features and stress states in a fcc material resulting from the S{sup 2}PD process.

  20. Surface Spins Pinning Effect on the Magnetic Properties in Co3O4 Nanocrystallites Covered with Polymer Decomposition Residues

    Institute of Scientific and Technical Information of China (English)

    李山东; 毕红; 方江邻; 钟伟; 都有为

    2004-01-01

    We prepare two kinds of Co3O4 antiferromagnetic nanocrystallite systems with different surface structures: one grain surface is covered by polymer decomposition residues (PDRs) (referred to as CS) and the other is naked (NS). It has been found that the magnetic properties of the CS sample deviate greatly from those of the NS sample. For example, the CS sample exhibits an open loop up to 8 T at 4.2K, while the two branches of the hysteresis loop for the NS sample superpose together when the field is in excess of 3 T. The average permanent magnetic moments per Co3O4 cell for the CS sample are about three times larger than that of the NS sample.The coercivity and loop shift for the CS sample are enhanced remarkably in comparison with the NS sample,i.e., from 73.8 and 11.0 kA/m for the NS sample to 116.5 and 25.5 kA/m for the CS sample, respectively. The anomalous magnetic properties and their enhancements for the CS sample are attributed to the surface spin pinning effect by PDRs.

  1. Detection of anionic energetic material residues in enhanced fingermarks on porous and non-porous surfaces using ion chromatography.

    Science.gov (United States)

    Love, Catherine; Gilchrist, Elizabeth; Smith, Norman; Barron, Leon

    2013-09-10

    The ability to link criminal activity and identity using validated analytical approaches can be of great value to forensic scientists. Herein, the factors affecting the recovery and detection of inorganic and organic energetic material residues within chemically or physically enhanced fingermarks on paper and glass substrates are presented using micro-bore anion exchange chromatography with suppressed conductivity detection. Fingermarks on both surfaces were enhanced using aluminium powder or ninhydrin after spiking with model test mixtures or through contact with black-powder substitutes. A quantitative study of the effects of environmental/method interferences, the sweat matrix, the surface and the enhancement technique on the relative anion recovery of forensically relevant species is presented. It is shown that the analytical method could detect target analytes at the nanogram level even within excesses of enhancement reagents and their reaction products when using solid phase extraction and/or microfiltration. To our knowledge, this work demonstrates for the first time that ion chromatography can detect anions in energetic materials within fingermarks on two very different surfaces, after operational enhancement techniques commonly used by forensic scientists and police have been applied.

  2. Mechanical and interfacial properties of poly(vinyl chloride) based composites reinforced by cassava stillage residue with different surface treatments

    Science.gov (United States)

    Zhang, Yanjuan; Gan, Tao; Li, Qian; Su, Jianmei; Lin, Ye; Wei, Yongzuo; Huang, Zuqiang; Yang, Mei

    2014-09-01

    Cassava stillage residue (CSR), a kind of agro-industrial plant fiber, was modified by coupling agent (CA), mechanical activation (MA), and MA-assisted CA (MACA) surface treatments, respectively. The untreated and different surface treated CSRs were used to prepare plant fibers/polymer composites (PFPC) with poly(vinyl chloride) (PVC) as polymer matrix, and the properties of these CSR/PVC composites were compared. Surface treated CSR/PVC composites possessed better mechanical properties, water resistance and dimensional stability compared with the untreated CSR/PVC composite, attributing to the improvement of interfacial properties between CSR and PVC matrix. MACA-treated CSR was the best reinforcement among four types of CSRs (untreated, MA-treated, CA-treated, and MACA-treated CSRs) because MACA treatment led to the significant improvement of dispersion, interfacial adhesion and compatibility between CSR and PVC. MACA treatment could be considered as an effective and green method for enhancing reinforcement efficiency of plant fibers and the properties of PFPC.

  3. Surface wipe sampling for antineoplastic (chemotherapy) and other hazardous drug residue in healthcare settings: Methodology and recommendations.

    Science.gov (United States)

    Connor, Thomas H; Zock, Matthew D; Snow, Amy H

    2016-09-01

    Surface wipe sampling for various hazardous agents has been employed in many occupational settings over the years for various reasons such as evaluation of potential dermal exposure and health risk, source determination, quality or cleanliness, compliance, and others. Wipe sampling for surface residue of antineoplastic and other hazardous drugs in healthcare settings is currently the method of choice to determine surface contamination of the workplace with these drugs. The purpose of this article is to review published studies of wipe sampling for antineoplastic and other hazardous drugs, to summarize the methods in use by various organizations and researchers, and to provide some basic guidance for conducting surface wipe sampling for these drugs in healthcare settings.  Recommendations on wipe sampling methodology from several government agencies and organizations were reviewed. Published reports on wipe sampling for hazardous drugs in numerous studies were also examined. The critical elements of a wipe sampling program and related limitations were reviewed and summarized.  Recommendations and guidance are presented concerning the purposes of wipe sampling for antineoplastic and other hazardous drugs in the healthcare setting, technical factors and variables, sampling strategy, materials required, and limitations. The reporting and interpretation of wipe sample results is also discussed.  It is recommended that all healthcare settings where antineoplastic and other hazardous drugs are handled consider wipe sampling as part of a comprehensive hazardous drug "safe handling" program. Although no standards exist for acceptable or allowable surface concentrations for these drugs in the healthcare setting, wipe sampling may be used as a method to characterize potential occupational dermal exposure risk and to evaluate the effectiveness of implemented controls and the overall safety program. A comprehensive safe-handling program for antineoplastic drugs may

  4. Dissolution and sorption of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and 2,4,6-trinitrotoluene (TNT) residues from detonated mineral surfaces.

    Science.gov (United States)

    Jaramillo, Ashley M; Douglas, Thomas A; Walsh, Marianne E; Trainor, Thomas P

    2011-08-01

    Composition B (Comp B) is a commonly used military formulation composed of the toxic explosive compounds 2,4,6-trinitrotoluene (TNT), and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). Numerous studies of the temporal fate of explosive compounds in soils, surface water and laboratory batch reactors have been conducted. However, most of these investigations relied on the application of explosive compounds to the media via aqueous addition and thus these studies do not provide information on the real world loading of explosive residues during detonation events. To address this we investigated the dissolution and sorption of TNT and RDX from Comp B residues loaded to pure mineral phases through controlled detonation. Mineral phases included nontronite, vermiculite, biotite and Ottawa sand (quartz with minor calcite). High Performance Liquid Chromatography and Attenuated Total Reflectance Fourier Transform Infrared spectroscopy were used to investigate the dissolution and sorption of TNT and RDX residues loaded onto the mineral surfaces. Detonation resulted in heterogeneous loading of TNT and RDX onto the mineral surfaces. Explosive compound residues dissolved rapidly (within 9 h) in all samples but maximum concentrations for TNT and RDX were not consistent over time due to precipitation from solution, sorption onto mineral surfaces, and/or chemical reactions between explosive compounds and mineral surfaces. We provide a conceptual model of the physical and chemical processes governing the fate of explosive compound residues in soil minerals controlled by sorption-desorption processes. Published by Elsevier Ltd.

  5. Study of the surface-enhanced Raman spectroscopy of residual impurities in hydroxylamine-reduced silver colloid and the effects of anions on the colloid activity.

    Science.gov (United States)

    Dong, Xiao; Gu, Huaimin; Liu, Fangfang

    2012-03-01

    The paper investigated the residual ions in hydroxylamine-reduced silver colloid (HRSC) and the relationship between the condition of HRSC and the enhanced mechanisms of this colloid. We also detected the SERS of MB and studied the effects of anions on the Raman signal. In the case of HRSC, the bands of residual ions diminish while the bands of Ag-anions increase gradually with increasing the concentrations of Cl(-) and NO(3)(-). It means the affinity of residual ions on the silver surface is weaker than that of Cl(-) and NO(3)(-) and the residual ions are replaced gradually by the added Cl(-) or NO(3)(-). The Raman signal of residual ions can be detected by treatment with anions that do not bind strongly to the silver surface, such as SO(4)(2-). The most intense band of Ag-anions bonds can be also observed when adding weakly binding anions to the colloid. However, the anions which make up the Ag-anions bonds are residual Cl(-) and the effect of weakly binding anions is only to aggregate the silver particles. Residual Cl(-) can be replaced by I(-) which has the highest affinity. From the detection of methylene blue (MB), the effects of anions on the enhancement of Raman signal are discussed in detail, and these findings could make the conditions suitable for detecting analytes in high efficiency. This study will have a profound implication to SERS users about their interpretation of SERS spectra when obtaining these anomalous bands.

  6. Detection of genotype-specific Ehrlichia canis exposure in Brazilian dogs by TRP36 peptide ELISA.

    Science.gov (United States)

    Aguiar, Daniel M; Zhang, Xiaofeng; Braga, Isis A; Taques, Isis I G G; McBride, Jere W

    2016-02-01

    We recently characterized a novel genotype of Ehrlichia canis based on the tandem repeat (TR) sequence of the TRP36 gene in Brazil. The TR amino acid sequence of the Brazilian (Br) genotype (ASVVPEAE) was divergent from the previously described US genotype (TEDSVSAPA) of E. canis. In this study, we developed an ELISA based on TRP36 TR synthetic peptides from both Br and US E. canis TRP36 genotypes to serologically detect and distinguish infections caused by these genotypes. Sera from 30 Brazilian dogs naturally infected with E. canis, sera from dogs experimentally infected E. canis (Jake and Cuiabá #1 strains) and E. chaffeensis (Arkansas strain) and 12 seronegative E. canis dogs were evaluated. Fifteen naturally infected Brazilian dogs had antibodies that reacted with the US TRP36 (n=9) or Br TRP36 (n=6) only, and 13 dogs had antibodies that reacted with both TPR36 peptides suggesting that these dogs were exposed to both genotypes. Most dogs (n=28) had antibodies that reacted with the highly conserved E. canis TRP19 peptide; however, two dogs had antibodies to E. canis TRP19, but did not have TRP36 antibodies, raising the possibility that another novel TRP36 genotype is circulating in Brazil. Our results demonstrate that synthetic peptides based on the TR region of E. canis TRP36 can be used to serologically distinguish infections or identify coinfections by different genotypes, and to determine the seroprevalence of various E. canis genotypes in Brazil.

  7. Mutations in Arabidopsis thaliana genes involved in the tryptophan biosynthesis pathway affect root waving on tilted agar surfaces

    Science.gov (United States)

    Rutherford, R.; Gallois, P.; Masson, P. H.

    1998-01-01

    Arabidopsis thaliana roots grow in a wavy pattern upon a slanted surface. A novel mutation in the anthranilate synthase alpha 1 (ASA1) gene, named trp5-2wvc1, and mutations in the tryptophan synthase alpha and beta 1 genes (trp3-1 and trp2-1, respectively) confer a compressed root wave phenotype on tilted agar surfaces. When trp5-2wvc1 seedlings are grown on media supplemented with anthranilate metabolites, their roots wave like wild type. Genetic and pharmacological experiments argue that the compressed root wave phenotypes of trp5-2wvc1, trp2-1 and trp3-1 seedlings are not due to reduced IAA biosynthetic potential, but rather to a deficiency in L-tryptophan (L-Trp), or in a L-Trp derivative. Although the roots of 7-day-old seedlings possess higher concentrations of free L-Trp than the shoot as a whole, trp5-2wvc1 mutants show no detectable alteration in L-Trp levels in either tissue type, suggesting that a very localized shortage of L-Trp, or of a L-Trp-derived compound, is responsible for the observed phenotype.

  8. Modulation of TRP channels by resveratrol and other stilbenoids

    Directory of Open Access Journals (Sweden)

    Yu Lina

    2013-02-01

    Full Text Available Abstract Background Resveratrol (3,5,4’ - trihydroxy-trans-stilbene, a widely distributed natural stilbenoid, was proposed to account for the unique effects of red wine on life span and health. It has been reported to possess various biological and pharmacological activities, such as anti-oxidant, anti-inflammatory, and anti-carcinogenic effects. Here, using whole-cell patch-clamp techniques and behavioral analyses, we investigated whether resveratrol and other stilbenoids can modulate TRP channels in sensory neurons in vitro, and have analgesic effects in vivo. Results We found that resveratrol dose-dependently suppressed the allyl isothiocyanate (AITC-induced currents (IAITC in HEK293 cells that express TRPA1, as well as in rat dorsal root ganglion (DRG neurons. Instead, pinosylvin methyl ether (PME, another derivate of stilbene which has a similar structure to resveratrol, dose-dependently blocked the capsaicin-induced currents (ICAP in HEK293 cells that express TRPV1 as well as in DRG neurons. Interestingly, resveratrol had no inhibitory effect on the ICAP, and PME had no effect on the IAITC. Otherwise, trans-stilbene showed no any effect on IAITC or ICAP. The concentration response curve of AITC showed that resveratrol inhibited the action of TRPA1 not by changing the EC50, but by suppressing the AITC-induced maximum response. By contrast, the inhibition of TRPV1 by PME did not change the capsaicin-induced maximum response but did cause a right shift of the EC50. Moreover, pre-administration of resveratrol suppressed intraplantar injections of AITC-evoked nocifensive behaviors, as well as that PME suppressed capsaicin-evoked one. Conclusions These data suggest that resveratrol and other stilbenoids may have an inhibitory effect on TRP channels. In addition, these stilbenoids modulate TRP channel activity in different ways.

  9. Ammonia volatilization from surface application of organic residues and urea on Marandu palisadegrass

    Directory of Open Access Journals (Sweden)

    Álisson Vanin

    2013-04-01

    Full Text Available The objective of this study was to measure ammonia volatilization from surface application of pig slurry, poultry litter, urea and no fertilization. An experiment was conducted in a randomized block design, in plots repeated over time, with four treatments and four replications. The fertilizers tested were: mineral fertilizer (70 kg ha-1 N, 100 kg ha-1 P2O5 and 30 kg ha-1 of K2O, as urea, triple superphosphate and potassium chloride, respectively, pig slurry (200 m³ ha-1 applied in November 2008 and 200 m³ ha-1 applied in April 2009 and poultry litter (10 t ha-1 applied in November 2008 and 10 t ha-1 applied in April 2009. Five evaluations were performed with 24-hour intervals. Foams were used with glycerin and sulfuric acid, internally fixed in PET bottles for collecting ammonia. After collected, the samples were sent to the laboratory to determine volatilized ammonia levels by the semi-micro Kjedahl distillation method. Fertilization with chicken litter had lower ammonia volatilization in relation to urea and pig slurry. Most of the ammonia volatilization from pig slurry occurred within 48 hours after application, totaling losses of 630 g kg-1 of a total of 8.25 kg ha-1 of volatilized ammonia. The biggest loss by ammonia volatilization was from fertilization with urea, totaling approximately 80 g kg-1 of N applied.

  10. Concerning the Optimal Salt Bridge for Trp-cage Stabilization†

    Science.gov (United States)

    Williams, D. Victoria; Byrne, Aimee; Stewart, James; Andersen, Niels H.

    2011-01-01

    Gai and co-workers (Bunagan, M. R. (2006) J. Phys. Chem. B 110, 3759-3763) reported computational design studies suggesting that a D9E mutation would stabilize the Trp-cage. Experimental studies for this mutation were reported in 2008 (Hudaky, P. (2008) Biochemistry 47, 1007-1016); the authors suggested that [D9E]-TC5b presented a more compact, and melting resistant structure due to the “optimal distance between the two sides of the molecule”. Nonetheless, the authors reported essentially the same CD melting temperature, 38±0.3 °C, for TC5b and its [D9E] mutant. In this study, a more stable Trp-cage, DAYAQ WLKDG GPSSG RPPPS, was examined by NMR and CD with the following mutations: [D9E], [D9R,R16E], [R16Orn], [D9E,R16Orn], [R16K], and [D9E,R16K]. Of these, the [D9E]-mutant displayed the smallest acidification induced change in the apparent Tm. In analogy to the prior study, the CD melts of TC10b and its [D9E] mutant were, however, very similar; all of the other mutations were significantly fold destabilizing by all measures. A detailed analysis indicates that the original D9/R16 salt bridge is optimal with regard to fold cooperativity and fold stabilization. Evidence for salt-bridging is also provided for a swapped pair, the [D9R,R16E]-mutant. Model systems reveal that an ionized aspartate at the C-terminus of a helix significantly decreases intrinsic helicity, a requirement for Trp-cage fold stability. The CD evidence which was cited as supporting increased fold stability for the [D9E]-TC5b at higher temperatures appears to be a reflection of increased helix stability in both the folded and unfolded state rather than a more favorable salt bridge. The present study also provides evidence for other Trp-cage stabilizing roles of the R16 sidechain. PMID:21222485

  11. Residual Distribution and Risk Assessment of Polychlorinated Biphenyls in Surface Sediments of the Pearl River Delta, South China.

    Science.gov (United States)

    Lai, Zini; Li, Xiuli; Li, Haiyan; Zhao, Lina; Zeng, Yanyi; Wang, Chao; Gao, Yuan; Liu, Qianfu

    2015-07-01

    We analyzed residual PCBs in surface sediments at 19 sites in the Pearl River Delta in the wet and dry seasons. Seven indicative PCB congeners (PCB28, PCB52, PCB101, PCB118, PCB153, PCB138 and PCB180) were detected in the surface sediments, among which the detection rate and mass concentrations of PCB52 were the highest. Total concentrations of the seven PCBs ranged from 19.8 to 111 μg/kg, with an average of 48.2 μg/kg. For the spatial distribution, the sum of the seven PCB (∑PCB) concentrations for the stations that were located in the city region of the Pearl River Delta were significantly higher than the ∑PCB concentrations for the eight outlets of the Pearl River Delta (p < 0.05). According to the US National Oceanic and Atmospheric Administration ERL and ERM guideline concentrations, the PCB concentrations may occasionally lead to adverse effects, especially in the dry season.

  12. Sampling the conformation of protein surface residues for flexible protein docking

    Directory of Open Access Journals (Sweden)

    Amenta Nina

    2010-11-01

    Full Text Available Abstract Background The problem of determining the physical conformation of a protein dimer, given the structures of the two interacting proteins in their unbound state, is a difficult one. The location of the docking interface is determined largely by geometric complementarity, but finding complementary geometry is complicated by the flexibility of the backbone and side-chains of both proteins. We seek to generate candidates for docking that approximate the bound state well, even in cases where there is backbone and/or side-chain difference from unbound to bound states. Results We divide the surfaces of each protein into local patches and describe the effect of side-chain flexibility on each patch by sampling the space of conformations of its side-chains. Likely positions of individual side-chains are given by a rotamer library; this library is used to derive a sample of possible mutual conformations within the patch. We enforce broad coverage of torsion space. We control the size of the sample by using energy criteria to eliminate unlikely configurations, and by clustering similar configurations, resulting in 50 candidates for a patch, a manageable number for docking. Conclusions Using a database of protein dimers for which the bound and unbound structures of the monomers are known, we show that from the unbound patch we are able to generate candidates for docking that approximate the bound structure. In patches where backbone change is small (within 1 Å RMSD of bound, we are able to account for flexibility and generate candidates that are good approximations of the bound state (82% are within 1 Å and 98% are within 1.4 Å RMSD of the bound conformation. We also find that even in cases of moderate backbone flexibility our candidates are able to capture some of the overall shape change. Overall, in 650 of 700 test patches we produce a candidate that is either within 1 Å RMSD of the bound conformation or is closer to the bound state than the

  13. [Role of thermo TRP channels in cutaneous neurogenic inflammation and itch].

    Science.gov (United States)

    XIE, Zhi-qiang

    2009-07-01

    The temperature-sensitive transient receptor potential (TRP) channels, is also called thermo TRP, including TRPV1, TRPV2, TRPV3, TRPV4, TRPM8 and TRPA1, which are expressed in sensory neurons and non-neuronal cells (e.g.keratinocyte, mast cell) of the skin. Thermo TRP channels are activated/sensitized by physical and chemical mediators, which participate in thermosensation and thermoregulation, so that they are key players in pruritus or pain pathogenesis. Thermo TRP channels are also involved in cutaneous neurogenic inflammation, thus they are regarded as molecular targets for future therapy in skin inflammation, pruritus and pain. In addition, following a basic syntax and molecular substrate of nociception and pruriception established by TRP channels-centered concept, the sensory categories can be distinguished and re-defined. Thermo TRP channels should be taken into account when analyzing the pathogenesis and management of itch or pruritic dermatosis.

  14. Seasonal and spatial variations of glyphosate residues in surface waters of El Crespo stream, Buenos Aires province, Argentina.

    Science.gov (United States)

    Perez, Debora; Okada, Elena; Aparicio, Virginia; Menone, Mirta; Costa, Jose Luis

    2017-04-01

    El Crespo stream is located inside a small watershed (52,000 Ha) which is only influenced by farming activities without urban or industrial impact. The watershed can be divided in two areas, the southern area (upstream), mainly composed of intensive crops and the northern area (downstream) used only for extensive livestock. In this sense, "El Crespo" stream in an optimal site for monitoring screening of pesticide residues. The objective of this work was to determine the seasonal and spatial variations of glyphosate (GLY), in surface waters of "El Crespo" stream. We hypothesized that in surface waters of "El Crespo" stream the levels of GLY vary depending of the season and rainfall events. The water sampling was carried out from October to June (2014-2015) in two sites: upstream (US) and downstream (DS), before and after rain events. The water samples were collected by triplicate in 1 L polypropylene bottles and stored at -20°C until analysis. GLY was extracted from unfiltered water samples with a buffer solution (100 mM Na2B4O7•10H2O/100 mM K3PO4, pH=9) and derivatized with 9-fluorenylmethylchloroformate (1 mg/mL in acetonitrile). Afterwards samples were analyzed using liquid chromatography coupled to a tandem mass spectrometer (UPLC-MS/MS). The detection limit (LD) was 0.1 μg/L and the quantification limit (QL) was 0.5 μg/L. The rainfall regime was obtained from the database of INTA Balcarce. GLY was detected in 92.3% of the analyzed samples. In the US site, were GLY is regularly applied, the highest GLY concentration was registered in October (2.15 ± 0.16 μg/L); from November to June, the GLY levels decreased from 1.97 ± 0.17 μg/L to residues found in October and November in both sites could be explained by the use of GLY in

  15. Caffeine and other methylxanthines as interceptors of food-borne aromatic mutagens: inhibition of Trp-P-1 and Trp-P-2 mutagenic activity.

    Science.gov (United States)

    Woziwodzka, Anna; Gołuński, Grzegorz; Wyrzykowski, Dariusz; Kaźmierkiewicz, Rajmund; Piosik, Jacek

    2013-11-18

    Caffeine is one of the most important biologically active food components. In this article, we demonstrate that caffeine and other methylxanthines significantly reduce the mutagenic activity of two food-derived heterocyclic aromatic amines, Trp-P-1 and Trp-P-2 in the Salmonella typhimurium TA98 strain. Moreover, protection against Trp-P-1-induced mutagenicity was independent of liver S9 enzymatic fraction, suggesting that mechanisms other than modulation of mutagen bioactivation can contribute to the observed protective effects. UV-vis spectroscopy and computational studies revealed that methylxanthines intercept Trp-P-1 and Trp-P-2 in noncovalent molecular complexes, with association constants (KAC) in the 10(2) M(-1) range. Enthalpy values (ΔH about -30 kJ·mol(-1)) of mutagen-methylxanthine heterocomplexation obtained microcalorimetrically correspond to stacking (π-π) interactions. Finally, we demonstrated that the biological activity of Trp-P-1 and Trp-P-2 is strictly dependent on the presence of the mutagen in a free (unbound with methylxanthine) form, suggesting that mutagen sequestration in stacking heterocomplexes with methylxanthines can decrease its bioavailability and diminish its biological effects.

  16. Glu-Trp-ONa or its acylated analogue (R-Glu-Trp-ONa) administration enhances the wound healing in the model of chronic skin wounds in rabbits

    Science.gov (United States)

    Shevtsov, Maxim A; Smagina, Larisa V; Kudriavtceva, Tatiana A; Petlenko, Sergey V; Voronkina, Irina V

    2015-01-01

    The management of chronic skin wounds represents a major therapeutic challenge. The synthesized dipeptide (Glu-Trp-ONa) and its acylated analogue (R-Glu-Trp-ONa) were assessed in the model of nonhealing dermal wounds in rabbits in relation to their healing properties in wound closure. Following wound modeling, the rabbits received a course of intraperitoneal injections of Glu-Trp-ONa or R-Glu-Trp-ONa. Phosphate-buffered saline and Solcoseryl® were applied as negative and positive control agents, respectively. An injection of Glu-Trp-ONa and R-Glu-Trp-ONa decreased the period of wound healing in animals in comparison to the control and Solcoseryl-treated groups. Acylation of Glu-Trp-ONa proved to be beneficial as related to the healing properties of the dipeptide. Subsequent zymography analyses showed that the applied peptides decreased the proteolytic activity of matrix metalloproteinases MMP-9, MMP-8, and MMP-2 in the early inflammatory phase and reversely increased the activity of MMP-9, MMP-8, and MMP-1 in the remodeling phase. Histological analyses of the wound sections (hematoxylin–eosin, Mallory’s staining) confirmed the enhanced formation of granulation tissue and re-epithelialization in the experimental groups. By administering the peptides, wound closures increased significantly through the modulation of the MMPs’ activity, indicating their role in wound healing. PMID:25848208

  17. Glu-Trp-ONa or its acylated analogue (R-Glu-Trp-ONa) administration enhances the wound healing in the model of chronic skin wounds in rabbits.

    Science.gov (United States)

    Shevtsov, Maxim A; Smagina, Larisa V; Kudriavtceva, Tatiana A; Petlenko, Sergey V; Voronkina, Irina V

    2015-01-01

    The management of chronic skin wounds represents a major therapeutic challenge. The synthesized dipeptide (Glu-Trp-ONa) and its acylated analogue (R-Glu-Trp-ONa) were assessed in the model of nonhealing dermal wounds in rabbits in relation to their healing properties in wound closure. Following wound modeling, the rabbits received a course of intraperitoneal injections of Glu-Trp-ONa or R-Glu-Trp-ONa. Phosphate-buffered saline and Solcoseryl® were applied as negative and positive control agents, respectively. An injection of Glu-Trp-ONa and R-Glu-Trp-ONa decreased the period of wound healing in animals in comparison to the control and Solcoseryl-treated groups. Acylation of Glu-Trp-ONa proved to be beneficial as related to the healing properties of the dipeptide. Subsequent zymography analyses showed that the applied peptides decreased the proteolytic activity of matrix metalloproteinases MMP-9, MMP-8, and MMP-2 in the early inflammatory phase and reversely increased the activity of MMP-9, MMP-8, and MMP-1 in the remodeling phase. Histological analyses of the wound sections (hematoxylin-eosin, Mallory's staining) confirmed the enhanced formation of granulation tissue and re-epithelialization in the experimental groups. By administering the peptides, wound closures increased significantly through the modulation of the MMPs' activity, indicating their role in wound healing.

  18. Pathways of electron transfer in Escherichia coli DNA photolyase: Trp306 to FADH.

    OpenAIRE

    1999-01-01

    We describe the results of a series of theoretical calculations of electron transfer pathways between Trp306 and *FADH. in the Escherichia coli DNA photolyase molecule, using the method of interatomic tunneling currents. It is found that there are two conformationally orthogonal tryptophans, Trp359 and Trp382, between donor and acceptor that play a crucial role in the pathways of the electron transfer process. The pathways depend vitally on the aromaticity of tryptophans and the flavin molecu...

  19. Ephemeral liquid water at the surface of the martian North Polar Residual Cap: Results of numerical modelling

    Science.gov (United States)

    Losiak, Anna; Czechowski, Leszek; Velbel, Michael A.

    2015-12-01

    Gypsum, a mineral that requires water to form, is common on the surface of Mars. Most of it originated before 3.5 Gyr when the Red Planet was more humid than now. However, occurrences of gypsum dune deposits around the North Polar Residual Cap (NPRC) seem to be surprisingly young: late Amazonian in age. This shows that liquid water was present on Mars even at times when surface conditions were as cold and dry as the present-day. A recently proposed mechanism for gypsum formation involves weathering of dust within ice (e.g., Niles, P.B., Michalski, J. [2009]. Nat. Geosci. 2, 215-220.). However, none of the previous studies have determined if this process is possible under current martian conditions. Here, we use numerical modelling of heat transfer to show that during the warmest days of the summer, solar irradiation may be sufficient to melt pure water ice located below a layer of dark dust particles (albedo ⩽ 0.13) lying on the steepest sections of the equator-facing slopes of the spiral troughs within martian NPRC. During the times of high irradiance at the north pole (every 51 ka; caused by variation of orbital and rotational parameters of Mars e.g., Laskar, J. et al. [2002]. Nature 419, 375-377.) this process could have taken place over larger parts of the spiral troughs. The existence of small amounts of liquid water close to the surface, even under current martian conditions, fulfils one of the main requirements necessary to explain the formation of the extensive gypsum deposits around the NPRC. It also changes our understanding of the degree of current geological activity on Mars and has important implications for estimating the astrobiological potential of Mars.

  20. Inter-Protein Sequence Co-Evolution Predicts Known Physical Interactions in Bacterial Ribosomes and the Trp Operon.

    Science.gov (United States)

    Feinauer, Christoph; Szurmant, Hendrik; Weigt, Martin; Pagnani, Andrea

    2016-01-01

    Interaction between proteins is a fundamental mechanism that underlies virtually all biological processes. Many important interactions are conserved across a large variety of species. The need to maintain interaction leads to a high degree of co-evolution between residues in the interface between partner proteins. The inference of protein-protein interaction networks from the rapidly growing sequence databases is one of the most formidable tasks in systems biology today. We propose here a novel approach based on the Direct-Coupling Analysis of the co-evolution between inter-protein residue pairs. We use ribosomal and trp operon proteins as test cases: For the small resp. large ribosomal subunit our approach predicts protein-interaction partners at a true-positive rate of 70% resp. 90% within the first 10 predictions, with areas of 0.69 resp. 0.81 under the ROC curves for all predictions. In the trp operon, it assigns the two largest interaction scores to the only two interactions experimentally known. On the level of residue interactions we show that for both the small and the large ribosomal subunit our approach predicts interacting residues in the system with a true positive rate of 60% and 85% in the first 20 predictions. We use artificial data to show that the performance of our approach depends crucially on the size of the joint multiple sequence alignments and analyze how many sequences would be necessary for a perfect prediction if the sequences were sampled from the same model that we use for prediction. Given the performance of our approach on the test data we speculate that it can be used to detect new interactions, especially in the light of the rapid growth of available sequence data.

  1. Metabolic activation of 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine and DNA adduct formation depends on p53: Studies in Trp53(+/+),Trp53(+/-) and Trp53(-/-) mice.

    Science.gov (United States)

    Krais, Annette M; Speksnijder, Ewoud N; Melis, Joost P M; Singh, Rajinder; Caldwell, Anna; Gamboa da Costa, Gonçalo; Luijten, Mirjam; Phillips, David H; Arlt, Volker M

    2016-02-15

    The expression of the tumor suppressor p53 can influence the bioactivation of, and DNA damage induced by, the environmental carcinogen benzo[a]pyrene, indicating a role for p53 in its cytochrome P450 (CYP)-mediated biotransformation. The carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), which is formed during the cooking of food, is also metabolically activated by CYP enzymes, particularly CYP1A2. We investigated the potential role of p53 in PhIP metabolism in vivo by treating Trp53(+/+), Trp53(+/-) and Trp53(-/-) mice with a single oral dose of 50 mg/kg body weight PhIP. N-(Deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP-C8-dG) levels in DNA, measured by liquid chromatography-tandem mass spectrometry, were significantly lower in liver, colon, forestomach and glandular stomach of Trp53(-/-) mice compared to Trp53(+/+) mice. Lower PhIP-DNA adduct levels in the livers of Trp53(-/-) mice correlated with lower Cyp1a2 enzyme activity (measured by methoxyresorufin-O-demethylase activity) in these animals. Interestingly, PhIP-DNA adduct levels were significantly higher in kidney and bladder of Trp53(-/-) mice compared to Trp53(+/+) mice, which was accompanied by higher sulfotransferase (Sult) 1a1 protein levels and increased Sult1a1 enzyme activity (measured by 2-naphthylsulfate formation from 2-naphthol) in kidneys of these animals. Our study demonstrates a role for p53 in the metabolism of PhIP in vivo, extending previous results on a novel role for p53 in xenobiotic metabolism. Our results also indicate that the impact of p53 on PhIP biotransformation is tissue-dependent and that in addition to Cyp1a enzymes, Sult1a1 can contribute to PhIP-DNA adduct formation.

  2. Water pressure head and temperature impact on isoxaflutole degradation in crop residues and loamy surface soil under conventional and conservation tillage management.

    Science.gov (United States)

    Alletto, Lionel; Coquet, Yves; Bergheaud, Valérie; Benoit, Pierre

    2012-08-01

    Laboratory incubations were performed in order to evaluate the dissipation of the proherbicide isoxaflutole in seedbed layer soil samples from conventional and conservation tillage systems and in maize and oat residues left at the soil surface under conservation tillage. The effects of temperature and water pressure head on radiolabelled isoxaflutole degradation were studied for each sample for 21d. Mineralisation of isoxaflutole was low for all samples and ranged from 0.0% to 0.9% of applied (14)C in soil samples and from 0.0% to 2.4% of applied (14)C in residue samples. In soil samples, degradation half-life of isoxaflutole ranged from 9 to 26h, with significantly higher values under conservation tillage. In residue samples, degradation half-life ranged from 3 to 31h, with significantly higher values in maize residues, despite a higher mineralisation and bound residue formation than in oat residues. Whatever the sample, most of the applied (14)C remained extractable during the experiment and, after 21d, less than 15% of applied (14)C were unextractable. This extractable fraction was composed of diketonitrile, benzoic acid derivative and several unidentified metabolites, with one of them accounting for more than 17% of applied (14)C. This study showed that tillage system design, including crop residues management, could help reducing the environmental impacts of isoxaflutole.

  3. Nucks1 synergizes with Trp53 to promote radiation lymphomagenesis in mice.

    Science.gov (United States)

    Yue, Yangbo; Leung, Stanley G; Liu, Yueyong; Huang, Yurong; Grundt, Kirsten; Østvold, Anne-Carine; Jen, Kuang-Yu; Schild, David; Mao, Jian-Hua; Wiese, Claudia

    2016-09-20

    NUCKS1 is a 27 kD vertebrate-specific protein, with a role in the DNA damage response. Here, we show that after 4 Gy total-body X-irradiation, Trp53+/- Nucks1+/- mice more rapidly developed tumors, particularly thymic lymphoma (TL), than Trp53+/- mice. TLs in both cohorts showed loss of heterozygosity (LOH) of the Trp53+ allele in essentially all cases. In contrast, LOH of the Nucks1+ allele was rare. Nucks1 expression correlated well with Nucks1 gene dosage in normal thymi, but was increased in the majority of TLs from Trp53+/- Nucks1+/- mice, suggesting that elevated Nucks1 message may be associated with progression towards malignancy in vivo. Trp53+/- Nucks1+/- mice frequently succumbed to CD4- CD8- TLs harboring translocations involving Igh but not Tcra/d, indicating TLs in Trp53+/- Nucks1+/- mice mostly originated prior to the double positive stage and at earlier lineage than TLs in Trp53+/- mice. Monoclonal rearrangements at Tcrb were more prevalent in TLs from Trp53+/- Nucks1+/- mice, as was infiltration of primary TL cells to distant organs (liver, kidney and spleen). We propose that, in the context of Trp53 deficiency, wild type levels of Nucks1 are required to suppress radiation-induced TL, likely through the role of the NUCKS1 protein in the DNA damage response.

  4. Role of TRP channels in the induction of heat shock proteins (Hsps) by heating skin.

    Science.gov (United States)

    Hsu, Wen-Li; Yoshioka, Tohru

    2015-01-01

    Transient receptor potential (TRP) channels in skin are crucial for achieving temperature sensitivity to maintain internal temperature balance and thermal homeostasis, as well as to protect skin cells from environmental stresses such as infrared (IR) or near-infrared (NIR) radiation via heat shock protein (Hsp) production. However, the mechanisms by which IR and NIR activate TRP channels and produce Hsps intracellularly have been independently reported. In this review, we discuss the relationship between TRP channel activation and Hsp production, and introduce the roles of several skin TRP channels in the regulation of HSP production by IR and NIR exposure.

  5. Comparative evaluation of persistence and residue of deltamethrin and cyfluthrin on different surfaces at Iranshahr area in Sistan & Baluchistan province in Iran 2004-2005

    Directory of Open Access Journals (Sweden)

    Seyed Mohammad Abtahi

    2007-02-01

    Full Text Available Background: Malaria is still one of the health problems in Iran. Resistance of Anopheles stephensi -the principal vector of the disease in the southeast of Iran- to insecticides such as DDT, malathion and dieldrin has been reported in south of the country. The present study was designed to estimate the persistence and residue of deltamethrin and cyfluthrin on different surfaces. Methods: After testing the surface of selected locations for previous insecticides, 4 locations were sprayed with deltamethrin (25 mg/m2 and cyfluthrin (20 mg/m2. A total of 22 conical tubes were installed and fixed on surfaces of walls and ceilings made from plaster, mud and wood. In each conical tube, 10 An. stephensi were released and after half an hour exposure these mosquitoes were transmitted to sterile cups. After 24 hours, the obtained results were recorded according to the survival and mortality rate of mosquitoes. These tests were carried out from 5 days after spraying and repeated every 15 days until the mortality rate decreased to 60-65%. Results: Persistence and residue rate of deltamethrin 5% with the scale of 25 mg/m2 of effective substance was 3 months at most. Afterwards persistence and residue effect of this insecticide decreased on absorbent and nonabsorbent surfaces. Persistence and residue rate of deltamethrin 10% with the scale of 20 mg/m2 on absorbent and nonabsorbent surfaces in Sistan & Baluchistan province climate was estimated 2.5 months. Afterwards, its persistence and residue effect decreased significantly. In the sprayed locations with deltamethrin, there was no correlation between the An. stephensi mortality and absorbent (plaster and muddy and nonabsorbent (wooden surfaces (P>0.05. Also, in locations sprayed with cyfluthrin there was no significant correlation between the two types of surfaces, however, the mortality rate was significantly different between plaster and muddy surfaces (P=0.007. Moreover, from the point of residual efficacy

  6. Combined Role of Two Tryptophane Residues of α-Factor Pheromone

    Energy Technology Data Exchange (ETDEWEB)

    Hong, Eun Young [Yeungnam Univ., Gyungsan (Korea, Republic of); Hong, Nam Joo [Seoul National Univ., Seoul (Korea, Republic of)

    2013-02-15

    Amide analogs of tridecapeptide α-factor (WHWLQLKPGQPMYCONH{sub 2}) of Saccharomyces cerevisiae, in which Trp at position 1 and 3 were replaced with other residues, were synthesized to ascertain whether cooperative interactions between two Trp residues occurred upon binding with its receptor. Analogs containing Ala or Aib at position 3 of the peptide [Ala{sup 3}]α-factor amide (2) and [Aib{sup 3}]α-factor amide (5) exhibited greater decreases in bioactivity than analogs with same residue at position one [Ala{sup 1}]α-factor amide (1) and [Aib{sup 1}]α-factor amide (4), reflecting that Trp{sup 3} may plays more important role than Trp{sup 1} for agonist activity. Analogs containing Ala or Aib in both position one and three 3, 6 exhibited complete loss of bioactivity, emphasizing both the essential role and the combined role of two indole rings for triggering cell signaling. In contrast, double substituted analog with D-Trp in both positions 9 exhibited greater activity than single substituted analog with D-Trp 8 or deleted analog 7, reflecting the combined contribution of two tryptophane residues of α-factor ligand to activation of Ste2p through interaction with residue Tyr{sup 266} and importance of the proper parallel orientation of two indole rings for efficient triggering of signal G protein coupled activation. Among ten amide analogs, [Ala{sup 1,3}]α-factor amide (3), [Aib{sup 1,3}]α-factor amide (6), [D-Trp{sup 3}]α-factor amide (8) and [des-Trp{sup 1},Phe{sup 3}]α-factor amide (10) were found to have antagonistic activity. Analogs 3 and 6 showed greater antagonistic activity than analogs 8 and 10.

  7. Surface mechanical property and residual stress of peened nickel-aluminum bronze determined by in-situ X-ray diffraction

    Science.gov (United States)

    Wang, Chengxi; Jiang, Chuanhai; Zhao, Yuantao; Chen, Ming; Ji, Vincent

    2017-10-01

    As one of the most important surface strengthening method, shot peening is widely used to improve the fatigue and stress corrosion crack resistance of components by introducing the refined microstructure and compressive residual stress in the surface layer. However, the mechanical properties of this thin layer are different from the base metal and are difficult to be characterized by conventional techniques. In this work, a micro uniaxial tensile tester equipped with in-situ X-ray stress analyzer was employed to make it achievable on a nickel-aluminum bronze with shot peening treatment. According to the equivalent stress-strain relationship based on Von Mises stress criterion, the Young's modulus and yield strength of the peened layer were calculated. The results showed that the Young's modulus was the same as the bulk material, and the yield strength corresponding to the permanent plastic strain of 0.2% was increased by 21% after SP. But the fractographic analysis showed that the fracture feature of the surface layer was likely to transform from the dimple to the cleavage, indicating the improved strength might be attained at the expense of ductility. The monotonic and cyclic loading were also performed via the same combined set-up. In addition, the specific relaxation behavior of compressive residual stress was quantified by linear logarithm relationship between residual stress and cycle numbers. It was found that the compressive residual stress mainly relaxed in the first few cycles, and then reached steady state with further cycles. The relaxation rate and the stable value were chiefly depended on the stress amplitude and number of cycles. The retained residual stress kept in compressive under all given applied stress levels, suggesting that the shot peening could introduce a more stable surface layer of compressive residual stress other than the elevated strength of nickel-aluminum bronze alloy.

  8. [Study on the Rapid Detection of Triazophos Residues in Flesh of Navel Orange by Using Surface-Enhanced Raman Scattering].

    Science.gov (United States)

    Wang, Xiao-bin; Wu, Rui-mei; Ling, Jing; Liu, Mu-hua; Zhang, Lu-ling; Lin, Lei; Chen, Jin-yin

    2016-03-01

    Surface enhanced Raman spectroscopy (SERS) and quick pre-treatment technology were used to detect triazophos residues in flesh of navel orange. Quantitative analysis model was developed by partial least squares (PLS) algorithm. SERS of different concentration (0.5 to 20 mg x L(-1)) triazophos juice solution with flesh extract as the matrix were collected by laser Raman spectrometer. Three preprocessing methods such as normalization, MSC and SNV were used to optimize Raman signals and PLS models were set up. The results showed that minimum detection concentration for triazophos in navel orange below 0.5 mg L(-1). The model built with normalization pre-processing gave the best result; the values of correlation (R(p)) and Root mean square error of prediction set (RMSEP) were 1.38 and 0.976 6, respectively. The predict recoveries were 95.97%-103.18%, and the absolute values of relative errors were below 5%. T-test (t = -0.018) showed that there was no significant difference between the true values and prediction values. This study demonstrates that this method is accurate and reliable.

  9. Pesticide management and their residues in sediments and surface and drinking water in the Mekong Delta, Vietnam.

    Science.gov (United States)

    Toan, Pham Van; Sebesvari, Zita; Bläsing, Melanie; Rosendahl, Ingrid; Renaud, Fabrice G

    2013-05-01

    Public concern in Vietnam is increasing with respect to pesticide pollution of the environment and of drinking water resources. While established monitoring programs in the Mekong Delta (MD) focus on the analysis of organochlorines and some organophosphates, the environmental concentrations of more recently used pesticides such as carbamates, pyrethroides, and triazoles are not monitored. In the present study, household level pesticide use and management was therefore surveyed and combined with a one year environmental monitoring program of thirteen relevant pesticides (buprofezin, butachlor, cypermethrin, α-endosulfan, β-endosulfan, endosulfan-sulfate, fenobucarb, fipronil, isoprothiolane, pretilachlor, profenofos, propanil, and propiconazole) in surface water, soil, and sediment samples. The surveys showed that household level pesticide management remains suboptimal in the Mekong Delta. As a consequence, a wide range of pesticide residues were present in water, soil, and sediments throughout the monitoring period. Maximum concentrations recorded were up to 11.24 μg l(-1) in water for isoprothiolane and up to 521 μg kg(-1) dm in sediment for buprofezin. Annual average concentrations ranged up to 3.34 μg l(-1) in water and up to 135 μg kg(-1) dm in sediment, both for isoprothiolane. Occurrence of pesticides in the environment throughout the year and co-occurrence of several pesticides in the samples indicate a considerable chronic exposure of biota and humans to pesticides. This has a high relevance in the delta as water for drinking is often extracted from canals and rivers by rural households (GSO, 2005, and own surveys). The treatment used by the households for preparing surface water prior to consumption (flocculation followed by boiling) is insufficient for the removal of the studied pesticides and boiling can actually increase the concentration of non-volatile pollutants.

  10. Efficacy and duration of three residual insecticides on cotton duck and vinyl tent surfaces for control of the sand fly Phlebotomus papatasi (Diptera: Psychodidae)

    Science.gov (United States)

    Many military tents are made of vinyl and cotton duck. Because it is useful to treat exterior tent surfaces to manage phlebotomine sand flies, DoD and ARS scientists evaluated the efficacy of 3 residual insecticides on both tent fabrics. P. papatasi was effectively killed by shade-stored and sun-exp...

  11. Role of lysine and tryptophan residues in the biological activity of toxin VII (Ts gamma) from the scorpion Tityus serrulatus.

    Science.gov (United States)

    Hassani, O; Mansuelle, P; Cestèle, S; Bourdeaux, M; Rochat, H; Sampieri, F

    1999-02-01

    Toxin VII (TsVII), also known as Ts gamma, is the most potent neurotoxin in the venom of the Brazilian scorpion Tityus serrulatus. It has been purified to homogeneity using a new fast and efficient method. Chemical modification of TsVII with the tryptophan-specific reagent o-nitrophenylsulfenyl chloride yielded three modified derivatives (residues Trp39, Trp50 and Trp54). Acetylation of TsVII mostly generated the monoacetylated Lys12 derivative. No side reactions were detected, as indicated by endoproteinase Lys-C peptide mapping, Edman degradation and electrospray mass spectrometry. Circular dichroism and fluorimetric measurements showed that none of the chemical modifications altered the overall structure of the derivatives. The acetylation of Lys12 or the sulfenylation of Trp39 or Trp54 led to a loss of both toxicity in mice and apparent binding affinity for rat brain and cockroach synaptosomal preparations. Sulfenylation of Trp50, however, moderately affected the toxicity of TsVII in mice and had almost no effect on its binding properties. A 3-dimensional model of TsVII was constructed by homology modeling. It suggests that the most reactive residues (Lys12 and Trp39 and Trp54) are all important in the functional disruption of neuronal sodium channels by TsVII, and are close to each other in the hydrophobic conserved region.

  12. Design of low surface roughness-low residual stress-high optoelectronic merit a-IZO thin films for flexible OLEDs

    Science.gov (United States)

    Kumar, Naveen; Wilkinson, Taylor M.; Packard, Corinne E.; Kumar, Mukesh

    2016-06-01

    The development of efficient and reliable large-area flexible optoelectronic devices demands low surface roughness-low residual stress-high optoelectronic merit transparent conducting oxide (TCO) thin films. Here, we correlate surface roughness-residual stress-optoelectronic properties of sputtered amorphous indium zinc oxide (a-IZO) thin films using a statistical design of experiment (DOE) approach and find a common growth space to achieve a smooth surface in a stress-free and high optoelectronic merit a-IZO thin film. The sputtering power, growth pressure, oxygen partial pressure, and RF/(RF+DC) are varied in a two-level system with a full factorial design, and results are used to deconvolve the complex growth space, identifying significant control growth parameters and their possible interactions. The surface roughness of a-IZO thin film varies over 0.19 nm to 3.97 nm, which is not in line with the general assumption of low surface roughness in a-IZO thin films. The initial regression model and analysis of variance reveal no single optimum growth sub-space to achieve low surface roughness (≤0.5 nm), low residual stress (-1 to 0 GPa), and industrially acceptable electrical conductivity (>1000 S/cm) for a-IZO thin films. The extrapolation of growth parameters in light of the current results and previous knowledge leads to a new sub-space, resulting in a low residual stress of -0.52±0.04 GPa, a low surface roughness of 0.55±0.03 nm, and moderate electrical conductivity of 1962±3.84 S/cm in a-IZO thin films. These results demonstrate the utility of the DOE approach to multi-parameter optimization, which provides an important tool for the development of flexible TCOs for the next-generation flexible organic light emitting diodes applications.

  13. Design of Low Surface Roughness-Low Residual Stress-High Optoelectronic Merit a-IZO Thin Films for Flexible OLEDs

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Naveen; Wilkinson, Taylor M.; Packard, Corinne E.; Kumar, Mukesh

    2016-06-14

    The development of efficient and reliable large-area flexible optoelectronic devices demands low surface roughness-low residual stress-high optoelectronic merit transparent conducting oxide (TCO) thin films. Here, we correlate surface roughness-residual stress-optoelectronic properties of sputtered amorphous indium zinc oxide (a-IZO) thin films using a statistical design of experiment (DOE) approach and find a common growth space to achieve a smooth surface in a stress-free and high optoelectronic merit a-IZO thin film. The sputtering power, growth pressure, oxygen partial pressure, and RF/(RF+DC) are varied in a two-level system with a full factorial design, and results are used to deconvolve the complex growth space, identifying significant control growth parameters and their possible interactions. The surface roughness of a-IZO thin film varies over 0.19 nm to 3.97 nm, which is not in line with the general assumption of low surface roughness in a-IZO thin films. The initial regression model and analysis of variance reveal no single optimum growth sub-space to achieve low surface roughness (=0.5 nm), low residual stress (-1 to 0 GPa), and industrially acceptable electrical conductivity (>1000 S/cm) for a-IZO thin films. The extrapolation of growth parameters in light of the current results and previous knowledge leads to a new sub-space, resulting in a low residual stress of -0.52 +/- 0.04 GPa, a low surface roughness of 0.55 +/- 0.03 nm, and moderate electrical conductivity of 1962 +/- 3.84 S/cm in a-IZO thin films. These results demonstrate the utility of the DOE approach to multi-parameter optimization, which provides an important tool for the development of flexible TCOs for the next-generation flexible organic light emitting diodes applications.

  14. Interplay between TRP channels and the cytoskeleton in health and disease.

    NARCIS (Netherlands)

    Clark, K.; Middelbeek, J.; Leeuwen, F.N. van

    2008-01-01

    Transient receptor potential (TRP) channels are a family of cation channels that play a key role in ion homeostasis and cell volume regulation. In addition, TRP channels are considered universal integrators of sensory information required for taste, vision, hearing, touch, temperature, and the detec

  15. Expression of transient receptor potential (TRP) channel mRNAs in the mouse olfactory bulb.

    Science.gov (United States)

    Dong, Hong-Wei; Davis, James C; Ding, ShengYuan; Nai, Qiang; Zhou, Fu-Ming; Ennis, Matthew

    2012-08-22

    Transient receptor potential (TRP) channels are a large family of cation channels. The 28 TRP channel subtypes in rodent are divided into 6 subfamilies: TRPC1-7, TRPV1-6, TRPM1-8, TRPP2/3/5, TRPML1-3 and TRPA1. TRP channels are involved in peripheral olfactory transduction. Several TRPC channels are expressed in unidentified neurons in the main olfactory bulb (OB), but the expression of most TRP channels in the OB has not been investigated. The present study employed RT-PCR as an initial survey of the expression of TRP channel mRNAs in the mouse OB and in 3 cell types: external tufted, mitral and granule cells. All TRP channel mRNAs except TRPV5 were detected in OB tissue. Single cell RT-PCR revealed that external tufted, mitral and granule cell populations expressed in aggregate 14 TRP channel mRNAs encompassing members of all 6 subfamilies. These different OB neuron populations expressed 7-12 channel mRNAs. Common channel expression was more similar among external tufted and mitral cells than among these cells and granule cells. These results indicate that a large number of TRP channel subtypes are expressed in OB neurons, providing the molecular bases for these channels to regulate OB neuron activity and central olfactory processing.

  16. Nociceptive TRP Channels: Sensory Detectors and Transducers in Multiple Pain Pathologies

    Directory of Open Access Journals (Sweden)

    Aaron D. Mickle

    2016-11-01

    Full Text Available Specialized receptors belonging to the transient receptor potential (TRP family of ligand-gated ion channels constitute the critical detectors and transducers of pain-causing stimuli. Nociceptive TRP channels are predominantly expressed by distinct subsets of sensory neurons of the peripheral nervous system. Several of these TRP channels are also expressed in neurons of the central nervous system, and in non-neuronal cells that communicate with sensory nerves. Nociceptive TRPs are activated by specific physico-chemical stimuli to provide the excitatory trigger in neurons. In addition, decades of research has identified a large number of immune and neuromodulators as mediators of nociceptive TRP channel activation during injury, inflammatory and other pathological conditions. These findings have led to aggressive targeting of TRP channels for the development of new-generation analgesics. This review summarizes the complex activation and/or modulation of nociceptive TRP channels under pathophysiological conditions, and how these changes underlie acute and chronic pain conditions. Furthermore, development of small-molecule antagonists for several TRP channels as analgesics, and the positive and negative outcomes of these drugs in clinical trials are discussed. Understanding the diverse functional and modulatory properties of nociceptive TRP channels is critical to function-based drug targeting for the development of evidence-based and efficacious new generation analgesics.

  17. Significance of the Centrally Expressed TRP Channel "Painless" in "Drosophila" Courtship Memory

    Science.gov (United States)

    Sakai, Takaomi; Sato, Shoma; Ishimoto, Hiroshi; Kitamoto, Toshihiro

    2013-01-01

    Considerable evidence has demonstrated that transient receptor potential (TRP) channels play vital roles in sensory neurons, mediating responses to various environmental stimuli. In contrast, relatively little is known about how TRP channels exert their effects in the central nervous system to control complex behaviors. This is also true for the…

  18. XPS characteristics of sulfur of bio-oxidized arsenic-bearing gold concentrate and changes of surface nature of bio-oxidation residue

    Institute of Scientific and Technical Information of China (English)

    杨洪英; 巩恩普; 杨立; 陈刚; 范有静; 张玉山; 吕久吉

    2004-01-01

    During bio-oxidation of sulfides, the chemical state change of sulfur is a complex and key factor. It is not only an indicator of the extent and intensity of the bio-oxidation, but also controls the property of bio-leaching medium and the period of oxidation. The chemical state of sulfur in sulfides oxidized by leaching bacteria was studied with XPS. Sulfide minerals in the arsenic-hearing gold concentrate consist of pyrite, arsenopyrite, chalcopyrite, galena, sphalerite and so on. In order to probe the pattern of the chemical state change of sulfur in the bio-oxidation residue of arsenic-bearing gold concentrate, the structure of the grains, and the surface nature of the residue, XPS test was carried out through different sputtering duration. The study of XPS clearly shows that: sulfides is progressively oxidized from the surface of minerals to the core by leaching bacteria; the chemical valence of sulfur changes from S2- or [S2]2- to [SO4]2-; sulfur in the core is in a reduction state, S2- or [S2]2- , but exists in an oxidation state S6+ on the surface; due to the chemical state change of sulfur, mineral phase of the bio-oxidation residue is also changed(sulfides inside, while sulfates outside); the layered structure is found in the grains of the bio-oxidation residue.

  19. A C. elegans sperm TRP protein required for sperm-egg interactions during fertilization.

    Science.gov (United States)

    Xu, X-Z Shawn; Sternberg, Paul W

    2003-08-08

    Fertilization, a critical step in animal reproduction, is triggered by a series of specialized sperm-egg interactions. However, the molecular mechanisms underlying fertilization are not well understood. Here, we identify a sperm-enriched C. elegans TRPC homolog, TRP-3. Mutations in trp-3 lead to sterility in both hermaphrodites and males due to a defect in their sperm. trp-3 mutant sperm are motile, but fail to fertilize oocytes after gamete contact. TRP-3 is initially localized in intracellular vesicles, and then translocates to the plasma membrane during sperm activation. This translocation coincides with a marked increase in store-operated calcium entry, providing an in vivo mechanism for the regulation of TRP-3 activity. As C. elegans oocytes lack egg coats, our data suggest that some TRPC family channels might function to mediate calcium influx during sperm-egg plasma membrane interactions leading to fertilization.

  20. In Vitro Repression of Transcription of the Trytophan Operon by trp Repressor

    Science.gov (United States)

    Shimizu, Yoshiko; Shimizu, Nobuyoshi; Hayashi, Masaki

    1973-01-01

    The in vitro repression of transcription of the tryptophan operon by the trp repressor from Escherichia coli was studied. By measuring the inhibitory effect for trp-specific RNA synthesis in an in vitro transcription system directed by DNA of trp-transducing phage, we have detected and concentrated a trp repressor in an eluate of a Φ80 ptED native DNA-cellulose column. The repression of transcription of trp operon required the addition of L-tryptophan in the system, and when several tryptophan analogues were added, the repression or derepression was similar to that observed in vivo. The repressor fraction was separated from the majority of tryptophanyl-tRNA synthetase activity by Bio-gel P60 column chromatography. PMID:4579009

  1. Thioredoxin-related protein 32 (TRP32) specifically reduces oxidized phosphatase of regenerating liver (PRL).

    Science.gov (United States)

    Ishii, Tasuku; Funato, Yosuke; Miki, Hiroaki

    2013-03-08

    PRL family constitutes a unique class of phosphatases associated with metastasis. The phosphatase activity of PRL has been reported to be important for promoting metastasis, and it is inactivated by reversible oxidation of its catalytic cysteine. Here, we show that TRP32 specifically reduces PRL. Reduction of oxidized PRL in cells is inhibited by 2,4-dinitro-1-chlorobenzene, an inhibitor of TRX reductase. In vitro assays for the reduction of PRL show that only TRP32 can potently reduce oxidized PRL, whereas other TRX-related proteins linked to TRX reductase show little or no reducing activity. Indeed, TRP32 knockdown significantly prolongs the H2O2-induced oxidation of PRL. Binding analyses reveal that the unique C-terminal domain of TRP32 is required and sufficient for its direct interaction with PRL. These results suggest that TRP32 maintains the reduced state of PRL and thus regulates the biological function of PRL.

  2. Functional interaction between TRP4 and CFTR in mouse aorta endothelial cells

    Directory of Open Access Journals (Sweden)

    Droogmans Guy

    2001-05-01

    Full Text Available Abstract Background This study describes the functional interaction between the putative Ca2+ channel TRP4 and the cystic fibrosis transmembrane conductance regulator, CFTR, in mouse aorta endothelium (MAEC. Results MAEC cells express CFTR transcripts as shown by RT-PCR analysis. Application of a phosphorylating cocktail activated a Cl- current with characteristics similar to those of CFTR mediated currents in other cells types (slow activation by cAMP, absence of rectification, block by glibenclamide. The current is present in trp4 +/+ MAEC, but not in trp4 -/- cells, although the expression of CFTR seems unchanged in the trp4 deficient cells as judged from RT-PCR analysis. Conclusions It is concluded that TRP4 is necessary for CFTR activation in endothelium, possibly by providing a scaffold for the formation of functional CFTR channels.

  3. Noncontact dipole effects on channel permeation. I. Experiments with (5F-indole)Trp13 gramicidin A channels.

    Science.gov (United States)

    Busath, D D; Thulin, C D; Hendershot, R W; Phillips, L R; Maughan, P; Cole, C D; Bingham, N C; Morrison, S; Baird, L C; Hendershot, R J; Cotten, M; Cross, T A

    1998-12-01

    Gramicidin A (gA), with four Trp residues per monomer, has an increased conductance compared to its Phe replacement analogs. When the dipole moment of the Trp13 side chain is increased by fluorination at indole position 5 (FgA), the conductance is expected to increase further. gA and FgA conductances to Na+, K+, and H+ were measured in planar diphytanoylphosphatidylcholine (DPhPC) or glycerylmonoolein (GMO) bilayers. In DPhPC bilayers, Na+ and K+ conductances increased upon fluorination, whereas in GMO they decreased. The low ratio in the monoglyceride bilayer was not reversed in GMO-ether bilayers, solvent-inflated or -deflated bilayers, or variable fatty acid chain monoglyceride bilayers. In both GMO and DPhPC bilayers, fluorination decreased conductance to H+ but increased conductance in the mixed solution, 1 M KCl at pH 2.0, where K+ dominates conduction. Eadie-Hofstee plot slopes suggest similar destabilization of K+ binding in both lipids. Channel lifetimes were not affected by fluorination in either lipid. These observations indicate that fluorination does not change the rotameric conformation of the side chain. The expected difference in the rate-limiting step for transport through channels in the two bilayers qualitatively explains all of the above trends.

  4. Chromium speciation and fractionation in ground and surface waters in the vicinity of chromite ore processing residue disposal sites.

    Science.gov (United States)

    Farmer, John G; Thomas, Rhodri P; Graham, Margaret C; Geelhoed, Jeanine S; Lumsdon, David G; Paterson, Edward

    2002-04-01

    Chromium concentrations of up to 91 mg l(-1) were found by ICP-OES for ground water from nine boreholes at four landfill sites in an area of S.E. Glasgow/S. Lanarkshire where high-lime chromite ore processing residue (COPR) from a local chemical works had been deposited from 1830 to 1968. Surface water concentrations of up to 6.7 mg l(-1) in a local tributary stream fell to 0.11 mg l(-1) in the River Clyde. Two independent techniques of complexation/colorimetry and speciated isotope dilution mass spectrometry (SIDMS) showed that Cr was predominantly (>90%) in hexavalent form (CrVI) as CrO4(2-), as anticipated at the high pH (7.5-12.5) of the sites. Some differences between the implied and directly determined concentrations of dissolved CrIII, however, appeared related to the total organic carbon (TOC) content. This was most significant for the ground water from one borehole that had the highest TOC concentration of 300 mg l(-1) and at which ultrafiltration produced significant decreases in Cr concentration with decreasing size fractions, e.g. complex. This showed for the main Cr-containing fraction, 100 kDa-0.45 microm, that the Cr was associated with a dark brown band characteristic of organic (humic) matter. Comparison of gel electrophoresis and FTIR results for ultrafilter retentates of ground water from this borehole with those for a borehole at another site where CrVI predominated suggested the influence of carboxylate groups, both in reducing CrVI and in forming soluble CrIII-humic complexes. The implications of this for remediation strategies (especially those based on the addition of organic matter) designed to reduce highly mobile and carcinogenic Cr(VI)O4(2-) to the much less harmful CrIII as insoluble Cr(OH)3 are discussed.

  5. Studies on the biotin-binding site of avidin. Tryptophan residues involved in the active site.

    Science.gov (United States)

    Gitlin, G; Bayer, E A; Wilchek, M

    1988-01-01

    Egg-white avidin was modified with the tryptophan-specific reagent 2-hydroxy-5-nitrobenzyl bromide. The complete loss of biotin-binding activity was achieved upon modification of an average of one tryptophan residue per avidin subunit. The identity of the modified residues was determined by isolating the relevant tryptic and chymotryptic peptides from CNBr-cleaved avidin fragments. The results demonstrate that Trp-70 and Trp-110 are modified in approximately equivalent proportions. It is believed that these residues are located in the active site of avidin and take part in the binding of biotin. PMID:3355517

  6. Studies on the biotin-binding site of avidin. Tryptophan residues involved in the active site.

    Science.gov (United States)

    Gitlin, G; Bayer, E A; Wilchek, M

    1988-02-15

    Egg-white avidin was modified with the tryptophan-specific reagent 2-hydroxy-5-nitrobenzyl bromide. The complete loss of biotin-binding activity was achieved upon modification of an average of one tryptophan residue per avidin subunit. The identity of the modified residues was determined by isolating the relevant tryptic and chymotryptic peptides from CNBr-cleaved avidin fragments. The results demonstrate that Trp-70 and Trp-110 are modified in approximately equivalent proportions. It is believed that these residues are located in the active site of avidin and take part in the binding of biotin.

  7. Effect of Plastic Pre-straining on Residual Stress and Composition Profiles in Low-Temperature Surface-Hardened Austenitic Stainless Steel

    Science.gov (United States)

    Bottoli, Federico; Christiansen, Thomas L.; Winther, Grethe; Somers, Marcel A. J.

    2016-08-01

    The present work deals with the evaluation of the residual stress profiles in expanded austenite by applying grazing incidence X-ray diffraction (GI-XRD) combined with successive sublayer removal. Annealed and deformed ( ɛ eq=0.5) samples of stable stainless steel EN 1.4369 were nitrided or nitrocarburized. The residual stress profiles resulting from the thermochemical low-temperature surface treatment were measured. The results indicate high-residual compressive stresses of several GPa's in the nitrided region, while lower-compressive stresses are produced in the carburized case. Plastic deformation in the steel prior to thermochemical treatment has a hardly measurable influence on the nitrogen-rich zone, while it has a measurable effect on the stresses and depth of the carbon-rich zone.

  8. Long-Term Residual Efficacy of Spinetoram on Concrete and Steel Surfaces for the Management of Three Stored Product Beetle Species.

    Science.gov (United States)

    Vassilakos, Thomas N; Athanassiou, Christos G

    2015-08-01

    In this study, the residual efficacy of spinetoram on concrete and galvanized steel surfaces was evaluated under fixed laboratory conditions against the rice weevil, Sitophilus oryzae (L.), the confused flour beetle, Tribolium confusum Jacquelin du Val, and the sawtoothed grain beetle, Oryzaephilus surinamensis (L.). Spinetoram was applied at the dose rates of 0.025 and 0.1 mg (active ingredient)/cm(2), on steel surfaces that were stored in continuous darkness and on concrete surfaces that were stored either in continuous darkness or in 12:12 (L:D) photoperiod. The experimental period for the residual effect of spinetoram was 6 mo. Bioassays were conducted for all types of surfaces and storage conditions at monthly intervals starting from the initial application period (seven bioassays in total). For each bioassay, mortality of the exposed adult beetles was measured after 3 and 7 d of exposure. Among the tested species, T. confusum was the least susceptible, regardless of the surface type, storage conditions, and dose rate. Regarding the bioassays conducted in the surfaces stored in darkness, spinetoram proved very persistent and no reduction in the efficacy was noted throughout the experimental period. Moreover, there were no differences in spinetoram efficacy between the two types of surfaces. Conversely, in light [12:12 (L:D)] conditions spinetoram efficacy was notably reduced after the first month, but remained stable for the rest of the period. The results of this study indicate that spinetoram was persistent with long residual efficacy against major stored grain beetle species on the most common types of surfaces in continuous darkness, while the presence of light reduced its efficacy. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  9. Glu-Trp-ONa or its acylated analogue (R-Glu-Trp-ONa administration enhances the wound healing in the model of chronic skin wounds in rabbits

    Directory of Open Access Journals (Sweden)

    Shevtsov MA

    2015-03-01

    Full Text Available Maxim A Shevtsov,1,2 Larisa V Smagina,1 Tatiana A Kudriavtceva,3 Sergey V Petlenko,4 Irina V Voronkina1 1Institute of Cytology of the Russian Academy of Sciences (RAS, St Petersburg, Russia; 2IP Pavlov State Medical University of St Petersburg, St Petersburg, Russia; 3Institute of Experimental Medicine of the North-West Branch of the Russian Academy of Medical Sciences (IEM NWB RAMS, St Petersburg, Russia; 4Military Medical Academy, St Petersburg, Russia Abstract: The management of chronic skin wounds represents a major therapeutic challenge. The synthesized dipeptide (Glu-Trp-ONa and its acylated analogue (R-Glu-Trp-ONa were assessed in the model of nonhealing dermal wounds in rabbits in relation to their healing properties in wound closure. Following wound modeling, the rabbits received a course of intraperitoneal injections of Glu-Trp-ONa or R-Glu-Trp-ONa. Phosphate-buffered saline and Solcoseryl® were applied as negative and positive control agents, respectively. An injection of Glu-Trp-ONa and R-Glu-Trp-ONa decreased the period of wound healing in animals in comparison to the control and Solcoseryl-treated groups. Acylation of Glu-Trp-ONa proved to be beneficial as related to the healing properties of the dipeptide. Subsequent zymography analyses showed that the applied peptides decreased the proteolytic activity of matrix metalloproteinases MMP-9, MMP-8, and MMP-2 in the early inflammatory phase and reversely increased the activity of MMP-9, MMP-8, and MMP-1 in the remodeling phase. Histological analyses of the wound sections (hematoxylin–eosin, Mallory’s staining confirmed the enhanced formation of granulation tissue and re-epithelialization in the experimental groups. By administering the peptides, wound closures increased significantly through the modulation of the MMPs’ activity, indicating their role in wound healing. Keywords: chronic wound, matrix metalloproteinases, small peptides

  10. Positions of Trp codons in the leader peptide-coding region of the at operon influence anti-trap synthesis and trp operon expression in Bacillus licheniformis.

    Science.gov (United States)

    Levitin, Anastasia; Yanofsky, Charles

    2010-03-01

    Tryptophan, phenylalanine, tyrosine, and several other metabolites are all synthesized from a common precursor, chorismic acid. Since tryptophan is a product of an energetically expensive biosynthetic pathway, bacteria have developed sensing mechanisms to downregulate synthesis of the enzymes of tryptophan formation when synthesis of the amino acid is not needed. In Bacillus subtilis and some other Gram-positive bacteria, trp operon expression is regulated by two proteins, TRAP (the tryptophan-activated RNA binding protein) and AT (the anti-TRAP protein). TRAP is activated by bound tryptophan, and AT synthesis is increased upon accumulation of uncharged tRNA(Trp). Tryptophan-activated TRAP binds to trp operon leader RNA, generating a terminator structure that promotes transcription termination. AT binds to tryptophan-activated TRAP, inhibiting its RNA binding ability. In B. subtilis, AT synthesis is upregulated both transcriptionally and translationally in response to the accumulation of uncharged tRNA(Trp). In this paper, we focus on explaining the differences in organization and regulatory functions of the at operon's leader peptide-coding region, rtpLP, of B. subtilis and Bacillus licheniformis. Our objective was to correlate the greater growth sensitivity of B. licheniformis to tryptophan starvation with the spacing of the three Trp codons in its at operon leader peptide-coding region. Our findings suggest that the Trp codon location in rtpLP of B. licheniformis is designed to allow a mild charged-tRNA(Trp) deficiency to expose the Shine-Dalgarno sequence and start codon for the AT protein, leading to increased AT synthesis.

  11. In various protein complexes, disordered protomers have large per-residue surface areas and area of protein-, DNA- and RNA-binding interfaces.

    Science.gov (United States)

    Wu, Zhonghua; Hu, Gang; Yang, Jianyi; Peng, Zhenling; Uversky, Vladimir N; Kurgan, Lukasz

    2015-09-14

    We provide first large scale analysis of the peculiarities of surface areas of 5658 dissimilar (below 50% sequence similarity) proteins with known 3D-structures that bind to proteins, DNA or RNAs. We show here that area of the protein surface is highly correlated with the protein length. The size of the interface surface is only modestly correlated with the protein size, except for RNA-binding proteins where larger proteins are characterized by larger interfaces. Disordered proteins with disordered interfaces are characterized by significantly larger per-residue areas of their surfaces and interfaces when compared to the structured proteins. These result are applicable for proteins involved in interaction with DNA, RNA, and proteins and suggest that disordered proteins and binding regions are less compact and more likely to assume extended shape. We demonstrate that disordered protein binding residues in the interfaces of disordered proteins drive the increase in the per residue area of these interfaces. Our results can be used to predict in silico whether a given protomer from the DNA, RNA or protein complex is likely to be disordered in its unbound form.

  12. Cleaning validation for residual estimation of olmesartan medoxomil on stainless steel surface of pharmaceutical manufacturing equipments using swab sampling and HPLC-DAD method

    Directory of Open Access Journals (Sweden)

    Nitin Dubey

    2013-06-01

    Full Text Available Prevention of cross contamination with active pharmaceutical ingredient is crucial and requires special attention in pharmaceutical industry. Current method validation describes residual determination of olmesartan medoxomil (OLME on stainless steel surface using swab sampling with a sensitive HPLC-DAD analysis. The acceptance limit was decided as 2 μg swab pro 100 cm2. Cotton swabs impregnated with extraction solution were used to determine residual drug content. Recoveries were 95.81%, 93.06%, and 96%. 24% with RSD below 1.5% at three concentration levels. Residual concentration was found to be linear in the range of 0.557–5.62 μg/mL, when estimated using Phenomenex Luna C18 (25 cm × 5 μm × 4.6 mm i.d. column at 1.0 mL/min flow rate at 258 nm. The mobile phase consisted of a mixture of acetonitrile: methanol: phosphate buffer pH 3.5: tetrahydrofuran (28:13:58:1 v/v/v/v. The LOD and LOQ for OLME were found to be 0.07 and 0.22 μg/mL, respectively. The validated method was found to be simple, selective and sensitive for demonstration of cleaning validation of OLME residues on the stainless steel surface.

  13. A novel strategy to analyze L-tryptophan through allosteric Trp repressor based on rolling circle amplification.

    Science.gov (United States)

    Zhao, Guojie; Hu, Tianyu; Li, Jun; Wei, Hua; Shang, Hong; Guan, Yifu

    2015-09-15

    Rolling circle amplification (RCA) has been considered as a powerful tool for nucleic acids detection. Here, a novel repressor-RCA-based method for L-tryptophan (L-Trp) detection was developed. This method utilizes the specific interaction between the RCA circular template and the Trp repressor protein (TrpR) involved in trp operon of Escherichia coli (E. coli). In the absence of L-Trp, the TrpR protein could not bind to the RCA template, and the RCA process can be continued. When L-Trp is present, the activated TrpR will bind to the operon sequence on the RCA template and inhibit the RCA reaction. Thus, the concentration of L-Trp is correlated directly with the fluorescent RCA signals. We succeeded in detecting L-Trp in a single step in simple homogeneous reaction system. The detection limit was estimated to be 0.77 μM (S/N=3) with good linearity. The method can unambiguously distinguish L-Trp from other 19 standard amino acids and L-Trp analogs. This strategy is also promising for detecting many small molecules such as other amino acids and carbohydrates.

  14. A structural view of ligand-dependent activation in thermoTRP channels

    Directory of Open Access Journals (Sweden)

    Ximena eSteinberg

    2014-05-01

    Full Text Available Transient Receptor Potential (TRP proteins are a large family of ion channels, grouped intoseven sub-families. Although great advances have been made regarding the activation andmodulation of TRP channel activity, detailed molecular mechanisms governing TRPchannel gating are still needed. Sensitive to electric, chemical, mechanical, and thermalcues, TRP channels are tightly associated with the detection and integration of sensoryinput, emerging as a model to study the polymodal activation of ion channel proteins.Among TRP channels, the temperature-activated kind constitute a subgroup by itself,formed by Vanilloid receptors 1-4, Melastatin receptors 2, 4, 5 and 8, TRPC5, and TRPA1.Some of the so-called thermoTRP channels participate in the detection of noxious stimulimaking them an interesting pharmacological target for the treatment of pain. However, thepoor specificity of the compounds available in the market represents an important obstacleto overcome. Understanding the molecular mechanics underlying ligand-dependentmodulation of TRP channels may help with the rational design of novel syntheticanalgesics. The present review focuses on the structural basis of ligand-dependentactivation of TRPV1 and TRPM8 channels. Special attention is drawn to the dissection ofligand-binding sites within TRPV1, PIP 2 -dependent modulation of TRP channels, and thestructure of natural and synthetic ligands.

  15. Probing Tertiary Structure of Proteins Using Single Trp Mutations with Circular Dichroism at Low Temperature

    Science.gov (United States)

    2015-01-01

    Trp is the most spectroscopically informative aromatic amino acid of proteins. However, the near-UV circular dichroism (CD) spectrum of Trp is complicated because the intensity and sign of 1La and 1Lb bands vary independently. To resolve vibronic structure and gain site-specific information from complex spectra, deconvolution was combined with cooling and site-directed tryptophan substitution. Low temperature near-UV CD was used to probe the local tertiary structure of a loop and α-helix in tear lipocalin. Upon cooling, the enhancement of the intensities of the near-UV CD was not uniform, but depends on the position of Trp in the protein structure. The most enhanced 1Lb band was observed for Trp at position 124 in the α-helix segment matching the known increased conformational mobility during ligand binding. Some aspects of the CD spectra of W28 and W130 were successfully linked to specific rotamers of Trp previously obtained from fluorescence lifetime measurements. The discussion was based on a framework that the magnitude of the energy differences in local conformations governs the changes in the CD intensities at low temperature. The Trp CD spectral classification of Strickland was modified to facilitate the recognition of pseudo peaks. Near-UV CD spectra harbor abundant information about the conformation of proteins that site directed Trp CD can report. PMID:24404774

  16. Role of Phe-99 and Trp-196 of sepiapterin reductase from Chlorobium tepidum in the production of L-threo-tetrahydrobiopterin

    Institute of Scientific and Technical Information of China (English)

    Supangat; Sun Ok Park; Kyung Hye Seo; Sang Yeol Lee; Young Shik Park; Kon Ho Lee

    2008-01-01

    Sepiapterin reductase from Chlorobium tepidum (cSR) catalyzes the synthesis of a distinct tetrahydrobiopterin (BH4), L-threo-BH4, different from the mammalian enzyme product. The 3-D crystal structure of cSR has revealed that the product configuration is determined solely by the substrate binding mode within the well-conserved catalytic triads. In cSR, the sepiapterin is stacked between two aromatic side chains of Phe-99 and Trp-196 and rotated approximately 180o around the active site from the position in mouse sepiapterin reductase. To confirm their roles in substrate binding, we mutated Phe-99 and/or Trp-196 to alanine (F99A, W196A) by site-directed mutagenesis and comparatively examined substrate binding of the purified proteins by kinetics analysis and differential scanning calorimetry. These mutants had higher Km values than the wild type. Remarkably, the W196A mutation resulted in a higher Km increase compared with the F99A mutation. Consistent with the results, the melting temperature (Tm) in the presence of sepiapterin was lower in the mutant proteins and the worst was W196A. These findings indicate that the two residues are indispensable for substrate binding in cSR, and Trp-196 is more important than Phe-99 for different stereoisomer production.

  17. Folding Thermodynamics and Mechanism of Five Trp-Cage Variants from Replica-Exchange MD Simulations with RSFF2 Force Field.

    Science.gov (United States)

    Zhou, Chen-Yang; Jiang, Fan; Wu, Yun-Dong

    2015-11-10

    To test whether our recently developed residue-specific force field RSFF2 can reproduce the mutational effect on the thermal stability of Trp-cage mini-protein and decipher its detailed folding mechanism, we carried out long-time replica-exchange molecular dynamics (REMD) simulations on five Trp-cage variants, including TC5b and TC10b. Initiated from their unfolded structures, the simulations not only well-reproduce their experimental structures but also their melting temperatures and folding enthalpies reasonably well. For each Trp-cage variant, the overall folding free energy landscape is apparently two-state, but some intermediate states can be observed when projected on more detailed coordinates. We also found different variants have the same major folding pathway, including the well formed PII-helix in the unfolded state, the formation of W6-P12/P18/P19 contacts and the α-helix before the transition state, the following formation of most native contacts, and the final native loop formation. The folding mechanism derived here is consistent with many previous simulations and experiments.

  18. Bacterial adherence in otitis media: Determination of N-acetylgalactosamine (GalNAc) residues in the submucosal glands and surface epithelium of the normal and diseased Eustachian tube

    DEFF Research Database (Denmark)

    Kirkeby, S; Friis, M; Mikkelsen, H B

    2011-01-01

    Acute otitis media (AOM) is the most common childhood infection caused by bacteria. The pathogenesis of AOM implicates initial adherence of a pathogen to the nasopharyngeal epithelium, which is followed by bacterial colonization of the middle ear cavity through the Eustachian tube. N......-acetylgalactosamine (GalNAc) is an important constituent of mucins and GalNAc containing sugar residues seem to be essential for initial adherence of respiratory bacteria to the surface of epithelial cells....

  19. Use of novel mutant galactosyltransferase for the bioconjugation of terminal N-Acetylglucosamine (GlcNAc) residues on live cell surface

    Science.gov (United States)

    Mercer, Natalia; Ramakrishnan, Boopathy; Boeggeman, Elizabeth; Verdi, Luke; Qasba, Pradman K.

    2013-01-01

    Based on the crystal structure of bovine β4Gal-T1 enzyme, mutation of a single amino acid Y289 to L289 (Y289L) changed its donor specificity from Gal to N-acetyl-galactosamine (GalNAc). A chemoenzymatic method that uses GalNAc analogues like GalNAz or 2-keto-Gal as sugar donors with the enzyme Y289LGal-T1 has identified hundreds of cytosolic and nuclear proteins that have O-GlcNAc modifications. To avoid potential cytotoxicity at Mn2+ concentrations required to selectively modify GlcNAc residues on the surface of live cells, we have engineered a Mg2+-dependent enzyme. Earlier, we have found that the mutation of the metal-binding residue Met-344 to His-344 in bovine β4Gal-T1 enzyme altered its meta-lion specificity in such a way that the M344H-β4Gal-T1 enzyme exhibits better catalytic activity with Mg2+ than with Mn2+. Here, we find that when these two mutations are combined, the double mutant, Y289L-M344H-β4Gal-T1, transfers GalNAc and its analogue sugars to the acceptor GlcNAc in the presence of Mg2+. Using this mutant enzyme, we have detected free GlcNAc residues on the surface glycans of live HeLa cells and platelets. The specific transfer of a synthetic sugar with a chemical handle to the terminal GlcNAc residues on the surface of live cells provides a novel tool for selective modification, detection, and isolation of GlcNAc-ending glycans present on the cellular surface. PMID:23259695

  20. Role of Ca++ Influx via Epidermal TRP Ion Channels

    Science.gov (United States)

    2014-10-01

    residual limb - prosthetic interface. Rather than attempting to enhance the functions of barrier-enhancing proteins, filaggrin (structural protein in...with considerable variation. For the lower strain, transcripts did not differ significantly. Human experimentation We finally addressed

  1. Label-Free and Multiplex Detection of Antibiotic Residues in Milk Using Imaging Surface Plasmon Resonance-Based Immunosensor

    NARCIS (Netherlands)

    Raz, Sabina Rebe; Bremer, Maria G. E. G.; Haasnoot, Willem; Norde, Willem

    2009-01-01

    Monitoring of antimicrobial drug residues in foods relies greatly on the availability of adequate analytical techniques. Currently, there is a need for a high-throughput screening method with a broad-spectrum detection range. This paper describes the development of a microarray biosensor, based on a

  2. Label-Free and Multiplex Detection of Antibiotic Residues in Milk Using Imaging surface Plasmon Resonance-Based immunosensor

    NARCIS (Netherlands)

    Rebe, S.; Bremer, M.G.E.G.; Haasnoot, W.; Norde, W.

    2009-01-01

    Monitoring of antimicrobial drug residues in foods relies greatly on the availability of adequate analytical techniques. Currently, there is a need for a high-throughput screening method with a broad-spectrum detection range. This paper describes the development of a microarray biosensor, based on a

  3. Brevicompanine C, cyclo-(D-Ile-L-Trp), and cyclo-(D-Leu-L-Trp), plant growth regulators from Penicillium brevi-compactum.

    Science.gov (United States)

    Kimura, Yasuo; Sawada, Aya; Kuramata, Masato; Kusano, Miyako; Fujioka, Shozo; Kawano, Tsuyoshi; Shimada, Atsumi

    2005-02-01

    New plant growth regulators, named brevicompanine C (1), cyclo-(D-Ile-L-Trp) (2), and cyclo-(D-Leu-L-Trp) (3), have been isolated from Penicillium brevi-compactum Dierckx, and their structures have been established by spectroscopic methods including 2D NMR and chiral TLC analysis. Plant growth activities of 1, 2, and 3 have been examined using lettuce seedling bioassay methods. All compounds accelerated the root growth of the seedlings in proportion to their concentration from 1 to 100 mg/L.

  4. Residues essential for Panton-Valentine leukocidin S component binding to its cell receptor suggest both plasticity and adaptability in its interaction surface.

    Directory of Open Access Journals (Sweden)

    Benoit-Joseph Laventie

    Full Text Available Panton-Valentine leukocidin (PVL, a bicomponent staphylococcal leukotoxin, is involved in the poor prognosis of necrotizing pneumonia. The present study aimed to elucidate the binding mechanism of PVL and in particular its cell-binding domain. The class S component of PVL, LukS-PV, is known to ensure cell targeting and exhibits the highest affinity for the neutrophil membrane (Kd∼10(-10 M compared to the class F component of PVL, LukF-PV (Kd∼10(-9 M. Alanine scanning mutagenesis was used to identify the residues involved in LukS-PV binding to the neutrophil surface. Nineteen single alanine mutations were performed in the rim domain previously described as implicated in cell membrane interactions. Positions were chosen in order to replace polar or exposed charged residues and according to conservation between leukotoxin class S components. Characterization studies enabled to identify a cluster of residues essential for LukS-PV binding, localized on two loops of the rim domain. The mutations R73A, Y184A, T244A, H245A and Y250A led to dramatically reduced binding affinities for both human leukocytes and undifferentiated U937 cells expressing the C5a receptor. The three-dimensional structure of five of the mutants was determined using X-ray crystallography. Structure analysis identified residues Y184 and Y250 as crucial in providing structural flexibility in the receptor-binding domain of LukS-PV.

  5. Cloning and sequencing of the trpE gene from Arthrobacter globiformis ATCC 8010 and several related subsurface Arthrobacter isolates

    Energy Technology Data Exchange (ETDEWEB)

    Chernova, T.; Viswanathan, V.K.; Austria, N.; Nichols, B.P.

    1998-09-01

    Tryptophan dependent mutants of Arthrobacter globiformis ATCC 8010 were isolated and trp genes were cloned by complementation and marker rescue of the auxotrophic strains. Rescue studies and preliminary sequence analysis reveal that at least the genes trpE, trpC, and trpB are clustered together in this organism. In addition, sequence analysis of the entire trpE gene, which encodes component I of anthranilate synthase, is described. Segments of the trpE gene from 17 subsurface isolates of Arthrobacter sp. were amplified by PCR and sequenced. The partial trpE sequences from the various strains were aligned and subjected to phylogenetic analysis. The data suggest that in addition to single base changes, recombination and genetic exchange play a major role in the evolution of the Arthrobacter genome.

  6. Site-specific uv crosslinking of minihelix DNA and TrpRS from Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    In order to investigate the recognition mechanism and the relationship between structure and function of minihelix DNA with Tryptophanyl-tRNA Synthetase (TrpRS), TrpRS from Bacillus Subtilis was purified. Four minihelix DNAs were chemically synthesized and the photoreactive reagent s4T was incorporated into three of them at the positions of G73, T72 and T55 corresponding to tRNATrp.The apparatus for uv crossiinking was devised and the parameters for uv crosslinking were optimized. The results indicated that the G73 and T72 base of minihelix DNA interacted with TrpRS directly. The uv crosslinking reaction was improved by the dose of uv irradiation and the concentration of both TrpRS and minihelix DNA.``

  7. Ingestion of TRP channel agonists attenuates exercise-induced muscle cramps.

    Science.gov (United States)

    Craighead, Daniel H; Shank, Sean W; Gottschall, Jinger S; Passe, Dennis H; Murray, Bob; Alexander, Lacy M; Kenney, W Larry

    2017-02-13

    Exercise associated muscle cramping (EAMC) is a poorly understood problem that is neuromuscular in origin. Ingestion of transient receptor potential (TRP) channel agonists has been efficacious in attenuating electrically-induced muscle cramps.

  8. Divergence of the TRP36 protein (gp36) in Ehrlichia canis strains found in Brazil.

    Science.gov (United States)

    Aguiar, Daniel M; Melo, Andreia L T

    2015-03-01

    The molecular characterization of 16S rRNA gene and immunoreactive proteins of Ehrlichia canis usually provide little information about the overall diversity of this organism. On the other hand, distinct sequences of the Tandem Repeat Protein 36 (TRP36/gp36) gene of E. canis have been reported, indicating substantial degree of diversity. The present letter aims to update and discuss the molecular divergence of the TRP36 protein between strains of E. canis isolated in different countries including Brazil.

  9. 铣削加工曲面残余应力有限元分析%Finite Element Analysis for Residual Stress of Milling Curved Surface

    Institute of Scientific and Technical Information of China (English)

    曾红; 韩笑; 陈燕燕; 王延忠; 吴林峰

    2013-01-01

      基于Abaqus软件对椭圆球面进行铣削加工仿真分析,通过热-力耦合的有限元理论对曲面表面进行残余应力分析,得到不同的加工参数下,表面残余应力的分布规律及变化规律,并通过正交试验得出影响残余应力的主要加工参数。%Elliptic spherical milling simulation analysis was made based on Abaqus software.Finite element theory of thermal-me-chanical coupling was used to make residual stress analysis for curved surface.Under different processing parameters,distribution and variation of surface residual stress were obtained.Through the orthogonal test,the main processing parameters affecting the residual stress were gotten.

  10. 金属表面处理磷化渣综合利用技术%Comprehensive Utilization of Phosphate Residues from Metal Surface Treatment

    Institute of Scientific and Technical Information of China (English)

    熊新宇; 熊天庆; 宋倩倩; 张诗萍

    2015-01-01

    The treatment methods for phosphate residues from metal surface treatment were studied. Phosphate residues from zinc-system phosphate solution was treated and recov-ered by adopting ammonia leaching complexing process to extract zinc phosphate, then adding sodium hydroxide into the residue to extract trisodium phosphate and ferric oxide or produce polymeric ferric sulfate water purifier. Industrial production has showed that it was a mature and convenient technology suitable for small metal surface treatment firms.%研究了金属表面处理所产生的磷化渣的处理技术,对锌系磷化液所产生的磷化渣,采用氨浸络合提取磷酸锌,残渣再加入烧碱提取磷酸三钠及氧化铁红或生产聚合硫酸铁净水剂进行综合利用。本项技术经工业规模的生产实践证明,技术成熟,操作简单,适合中小型金属表面处理企业应用。

  11. Adsorption of vapreotide on gold colloids studied by surface enhanced Raman spectroscopy

    Science.gov (United States)

    Gómez, J. A.; Cabanzo, R.; Mejia Ospino, E.

    2016-02-01

    Surface Enhanced Raman Spectroscopy (SERS) has been used to investigate the somatostatin (SST) analogue Vapreotide (VAP) in gold colloids. The optimum conditions to detect SERS signals of VAP have been studied. The observed SERS bands correspond to different vibrational modes of the peptide; being the most dominant SERS signals the ones derived from the aromatic amino acids Tryptophan (Trp), Phenylalanine (Phe) and Tyrosine (Tyr). Changes in enhancement and wavenumber of the proper bands upon adsorption on gold colloid are consistent with VAP adsorption, primarily through Tryptophan residues.

  12. Dual Loss of Rb1 and Trp53 in the Adrenal Medulla Leads to Spontaneous Pheochromocytoma

    Directory of Open Access Journals (Sweden)

    Ian D. Tonks

    2010-03-01

    Full Text Available Using a Cre/loxP system, we have determined the phenotypic consequences attributable to in vivo deletion of both Rb1 and Trp53 in the mouse adrenal medulla. The coablation of these two tumor suppressor genes during embryogenesis did not disrupt adrenal gland development but resulted in the neoplastic transformation of the neural crest-derived adrenal medulla, yielding pheochromocytomas (PCCs that developed with complete penetrance and were inevitably bilateral. Despite their typically benign status, these PCCs had profound ramifications on mouse vitality, with effected mice having a median survival of only 121 days. Evaluation of these PCCs by both immunohistochemistry and electron microscopy revealed that most Rb1-/-:Trp53-/- chromaffin cells possessed atypical chromagenic vesicles that did not seem capable of appropriately storing synthesized catecholamines. The structural remodeling of the heart in mice harboring Rb1-/-:Trp53-/- PCCs suggests that the mortality of these mice may be attributable to the inappropriate release of catecholamines from the mutated adrenal chromaffin cells. On the basis of the collective data from Rb1 and Trp53 knockout mouse models, it seems that the conversion of Rb1 loss-driven adrenal medulla hyperplasia to PCC can be greatly enhanced by the compound loss of Trp53, whereas the loss of Trp53 alone is generally ineffectual on adrenal chromaffin cell homeostasis. Consequently, the Trp53 tumor suppressor gene is an efficient genetic modifier of Rb1 loss in the development of PCC, and their compound loss in the adrenal medulla has a profound impact on both cellular homeostasis and animal vitality.

  13. BAX and tumor suppressor TRP53 are important in regulating mutagenesis in spermatogenic cells in mice.

    Science.gov (United States)

    Xu, Guogang; Vogel, Kristine S; McMahan, C Alex; Herbert, Damon C; Walter, Christi A

    2010-12-01

    During the first wave of spermatogenesis, and in response to ionizing radiation, elevated mutant frequencies are reduced to a low level by unidentified mechanisms. Apoptosis is occurring in the same time frame that the mutant frequency declines. We examined the role of apoptosis in regulating mutant frequency during spermatogenesis. Apoptosis and mutant frequencies were determined in spermatogenic cells obtained from Bax-null or Trp53-null mice. The results showed that spermatogenic lineage apoptosis was markedly decreased in Bax-null mice and was accompanied by a significantly increased spontaneous mutant frequency in seminiferous tubule cells compared to that of wild-type mice. Apoptosis profiles in the seminiferous tubules for Trp53-null were similar to control mice. Spontaneous mutant frequencies in pachytene spermatocytes and in round spermatids from Trp53-null mice were not significantly different from those of wild-type mice. However, epididymal spermatozoa from Trp53-null mice displayed a greater spontaneous mutant frequency compared to that from wild-type mice. A greater proportion of spontaneous transversions and a greater proportion of insertions/deletions 15 days after ionizing radiation were observed in Trp53-null mice compared to wild-type mice. Base excision repair activity in mixed germ cell nuclear extracts prepared from Trp53-null mice was significantly lower than that for wild-type controls. These data indicate that BAX-mediated apoptosis plays a significant role in regulating spontaneous mutagenesis in seminiferous tubule cells obtained from neonatal mice, whereas tumor suppressor TRP53 plays a significant role in regulating spontaneous mutagenesis between postmeiotic round spermatid and epididymal spermatozoon stages of spermiogenesis.

  14. Nanostructure of Poly(Acrylic Acid) Adsorption Layer on the Surface of Activated Carbon Obtained from Residue After Supercritical Extraction of Hops

    Science.gov (United States)

    Wiśniewska, M.; Nosal-Wiercińska, A.; Ostolska, I.; Sternik, D.; Nowicki, P.; Pietrzak, R.; Bazan-Wozniak, A.; Goncharuk, O.

    2017-01-01

    The nanostructure of poly(acrylic acid) (PAA) adsorption layer on the surface of mesoporous-activated carbon HPA obtained by physical activation of residue after supercritical extraction of hops was characterized. This characterization has been done based on the analysis of determination of adsorbed polymer amount, surface charge density, and zeta potential of solid particles (without and in the PAA presence). The SEM, thermogravimetric, FTIR, and MS techniques have allowed one to examine the solid surface morphology and specify different kinds of HPA surface groups. The effects of solution pH, as well as polymer molecular weight and concentration, were studied. The obtained results indicated that the highest adsorption on the activated carbon surface was exhibited by PAA with lower molecular weight (i.e., 2000 Da) at pH 3. Under such conditions, polymeric adsorption layer is composed of nanosized PAA coils (slightly negatively charged) which are densely packed on the positive surface of HPA. Additionally, the adsorption of polymeric macromolecules into solid pores is possible.

  15. Assessment of a handheld fluorescence imaging device as a visual-aid for detection of food residues on processing surfaces

    Science.gov (United States)

    Contamination of food with pathogenic bacteria can lead to foodborne illnesses. Food processing surfaces can serve as a medium for cross-contamination if sanitization procedures are inadequate. Ensuring that food processing surfaces are correctly cleaned and sanitized is important in the food indust...

  16. EFFECT OF TENSILE STRESS AND RESIDUAL STRESS ON THE SPONTANEOUS STRAY FIELD SIGNALS FROM THE SURFACE OF 0.45%C STEEL

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    In order to explore the quantitative method of metal magnetic memory testing(MMMT) and clarify the relationship between Hp(y), the normal component of spontaneous stray field, and applied stress or residual stress, the static tensile tests of 0.45%C steel sheet specimens are carried out on a servo hydraulic MTS810 machine. Hp(y) values are measured during the test process by an EMS-2003 metal magnetic memory diagnostic apparatus and a non-magnetic electric control displacement instrument. Residual stresses of some points on the surface of a specimen are measured by a Stress Tech X-Stress 3000 X-ray diffraction instrument. The results show that the same variation rules of Hp(y) value versus applied tensile stress are presented under the different conditions of load-on and load-off. However, the same rule does not exist between the Hp(y) value and residual stress. The variation of Hp(y) value reflects the history of applied tensile stress.

  17. Employing the fluorescence anisotropy and quenching kinetics of tryptophan to hunt for residual structures in denatured proteins

    Indian Academy of Sciences (India)

    Satish Kumar; Rajaram Swaminathan

    2007-03-01

    Residual structures in denatured proteins have acquired importance in recent years owing to their role as protein-folding initiation sites. Locating these structures in proteins has proved quite formidable, requiring techniques like NMR. Here in this report, we take advantage of the ubiquitous presence of tryptophan residues in residual structures to hunt for their presence using steady-state fluorescence spectroscopy. The surface accessibility and rotational dynamics of tryptophan in putative residual structures among ten different proteins, namely glucagon, melittin, subtilisin carlsberg, myelin basic protein, ribonuclease T1, human serum albumin, barstar mutant, bovine serum albumin, lysozyme and Trp-Met-Asp-Phe-NH2 peptide, was studied using steady state fluorescence quenching and anisotropy, respectively. Five proteins, namely ribonuclease T1, bovine serum albumin, melittin, barstar and hen egg white lysozyme appear likely to possess tryptophan(s) in hydrophobic clusters based on their reduced bimolecular quenching rates and higher steady-state anisotropy in proportion to their chain length. We also show that the fluorescence emission maximum of tryptophan is insensitive to the presence of residual structures.

  18. BSA adsorption on aliphatic and aromatic acid SAMs: investigating the effect of residual surface charge and sublayer nature.

    Science.gov (United States)

    Vallée, Anne; Humblot, Vincent; Al Housseiny, Rana; Boujday, Souhir; Pradier, Claire-Marie

    2013-09-01

    In this work, the influence of surface charge and layer rigidity on Bovin Serum Albumin (BSA) adsorption has been investigated. To this aim, Self Assembled Monolayers (SAMs) bearing terminal COOH or COO(-) groups were built on gold surfaces. The rigidity of the acid terminated SAMs was modified using either an aliphatic, mercaptoundecanoic acid (MUA), or an aromatic, mercaptobenzoic acid (MBA) thiol. X-Ray Photoelectron Spectroscopy (XPS), Polarization Modulation Reflection Absorption Infrared Spectroscopy (PM-RAIRS) and contact angle measurements, were used to deeply characterize the so-built layers. The surface charge was successfully modified by varying the pH of the rinsing solution. Indeed, COOH were the dominating species upon rinsing at pH 2 and COO(-) species dominated upon rinsing at pH 11. Rinsing at an intermediate pH, 5.5, led to the coexistence of both carboxylic and carboxylate moieties. The hydrophilic character of the surface was also found to depend on the rinsing pH, with a minimum after rinsing at intermediate pH. Using aromatic or aliphatic thiols did not affect the speciation but led to considerable differences in the hydrophilic character of these surfaces. Eventually, the adsorption of BSA on the acidic layers was investigated using PM-RAIRS. The results showed interesting differences between the charged layers. Thus, for both MUA and MBA -based SAMs, the amount of adsorbed proteins decreased when the amount of COO(-) on the surface increased. Interestingly, these effects were totally annihilated when the adsorption was carried out in PBS buffer. Moreover, for similar surface charges, the aromatic layers were able to bind higher amounts of proteins than the aliphatic ones. This work points out the key role of both surface charge and rigidity on protein adsorption. The influence of additional parameters, such as hydrophilicity and SAMs' rigidity is also established.

  19. Cloning of TRP Gene of Bombyx mori%家蚕TPR基因的克隆

    Institute of Scientific and Technical Information of China (English)

    牛伟涛

    2011-01-01

    [ Objective ] The aim was to clone Bornbyx rnori TRP gene. [ Method ] The total RNA of Bombyx mori was extracted during its pupal pedod with Trizol method, and then the TRP gene was cloned by RT-PCR. [ Result] The full-length cDNA of TRP gene was successfully obtained, and the ORF fragment (858 bp) of TRP gene was cloned by PCR. [ Conclusion] TRP gene of Bombyx mori was cloned for the first time,which could provide solid basis for the study on its function.%[目的]克隆家蚕TPR基因,为研究该基因功能提供依据.[方法]利用Trizol法提取家蚕蛹期总RNA,再用RT-PCR的方法对家蚕TPR基因进行克隆.[结果]成功得到家蚕TPR基因全cDNA序列,并用PCR扩增得到了TPR基因的ORF,全长858 bp.[结论]首次对家蚕TPR家族基因进行了克隆,为今后对该基因功能研究奠定了坚实的基础.

  20. Cloning of TRP Gene of Bombyx mori%家蚕TPR基因的克隆

    Institute of Scientific and Technical Information of China (English)

    牛伟涛

    2011-01-01

    [ Objective ] The aim was to clone Bombyx mori TRP gene. [ Method ] The total RNA of Bombyx mori was extracted during its pupal period with Trizol method, and then the TRP gene was cloned by RT-PCR. [ Result] The full-length cDNA of TRP gene was successfully obtained, and the ORF fragment (858 bp) of TRP gene was cloned by PCR. [Conclusion] TRP gene of Bombyx mori was cloned for the first time, which could provide solid basis for the study on its function.%[目的]克隆家蚕TPR基因,为研究该基因功能提供依据.[方法]利用Trizol法提取家蚕蛹期总RNA,再用RT-PCR方法对家蚕TPR基因进行克隆.[结果]成功得到家蚕TPR基因全cDNA序列,并用PCR扩增得到了TPR基因的ORF,全长858 bp.[结论]首次对家蚕TPR家族基因进行了克隆,为今后对该基因功能研究奠定了坚实的基础.

  1. Evolution of TRP channels inferred by their classification in diverse animal species.

    Science.gov (United States)

    Peng, Guangda; Shi, Xiao; Kadowaki, Tatsuhiko

    2015-03-01

    The functions of TRP channels have primarily been characterized in model organisms within a limited evolutionary context. We thus characterize the TRP channels in choanoflagellate, sponge, Cnidaria, Lophotrochozoa, and arthropods to understand how they emerged during early evolution of animals and have changed during diversification of various species. As previously reported, five metazoan TRP subfamily members (TRPA, TRPC, TRPM, TRPML, and TRPV) were identified in choanoflagellates, demonstrating that they evolved before the emergence of multicellular animals. TRPN was identified in Hydra magnipapillata, and therefore emerged in the last common ancestor of Cnidaria-Bilateria. A novel subfamily member (TRPVL) was identified in Cnidaria and Capitella teleta, indicating that it was present in the last common ancestor of Cnidaria-Bilateria but has since been lost in most bilaterians. The characterization of arthropod TRP channels revealed that Daphnia pulex and insects have specifically expanded the TRPA subfamily, which diverged from the ancient TRPA1 channel gene. The diversity of TRPA channels except TRPA1 was detectable even within a single insect family, namely the ant lineage. The present study demonstrates the evolutionary history of TRP channel genes, which may have diverged in conjunction with the specific habitats and life histories of individual species. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. POTENTIAL APPLICATIONS OF SOS-GFP BIOSENSOR TO IN VITRO RAPID SCREENING OF CYTOTOXIC AND GENOTOXIC EFFECT OF ANTICANCER AND ANTIDIABETIC PHARMACIST RESIDUES IN SURFACE WATER

    Directory of Open Access Journals (Sweden)

    Marzena Matejczyk

    2014-12-01

    Full Text Available Escherichia coli K-12 GFP-based bacterial biosensors allowed the detection of cytotoxic and genotoxic effect of anticancer drug– cyclophosphamide and antidiabetic drug – metformin in PBS buffer and surface water. Experimental data indicated that recA::gfpmut2 genetic system was sensitive to drugs and drugs mixture applied in experiment. RecA promoter was a good bioindicator in cytotoxic and genotoxic effect screening of cyclophosphamide, metformin and the mixture of the both drugs in PBS buffer and surface water. The results indicated that E. coli K-12 recA::gfp mut2 strain could be potentially useful for first-step screening of cytotoxic and genotoxic effect of anticancer and antidiabetic pharmacist residues in water. Next steps in research will include more experimental analysis to validate recA::gfpmut2 genetic system in E. coli K-12 on different anticancer drugs.

  3. Application of comprehensive two-dimensional gas chromatography with mass spectrometric detection for the analysis of selected drug residues in wastewater and surface water

    Institute of Scientific and Technical Information of China (English)

    Petr Lacina; Ludmila Mravcová; Milada Vávrová

    2013-01-01

    Pharmaceutical residues have become tightly controlled environmental contaminants in recent years,due to their increasing concentration in environmental components.This is mainly caused by their high level of production and everyday consumption.Therefore there is a need to apply new and sufficiently sensitive analytical methods,which can detect the presence of these contaminants even in very low concentrations.This study is focused on the application of a reliable analytical method for the analysis of 10 selected drug residues,mainly from the group of non-steroidal anti-iaffammatory drugs (salicylic acid,acetylsalicylic acid,clofibric acid,ibuprofen,acetaminophen,caffeine,naproxen,mefenamic acid,ketoprofen,and dicofenac),in wastewaters and surface waters.This analytical method is based on solid phase extraction,derivatization by N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) and finally analysis by comprehensive two-dimensional gas chromatography with Time-of-Flight mass spectrometric detection (GCxGC-TOF MS).Detection limits ranged from 0.18 to 5 ng/L depending on the compound and selected matrix.The method was successfully applied for detection of the presence of selected pharmaceuticals in the Svratka River and in wastewater from the wastewater treatment plant in Brno-Modrice,Czech Republic.The concentration of pharmaceuticals varied from one to several hundreds of ng/L in surface water and from one to several tens of μg/L in wastewater.

  4. Effects of Different Surfaces and Insecticide Carriers on Residual Insecticide Bioassays Against Bed Bugs, Cimex spp. (Hemiptera: Cimicidae).

    Science.gov (United States)

    Dang, Kai; Singham, G Veera; Doggett, Stephen L; Lilly, David G; Lee, Chow-Yang

    2017-04-01

    The performance of five insecticides (bendiocarb, deltamethrin, DDT, malathion, and imidacloprid) using three application methods (oil-based insecticide films on filter paper, and acetone-based insecticide deposits on two substrates: filter paper and glass) was assessed against a susceptible strain of Cimex lectularius (L.) and two resistant strains of Cimex hemipterus (F.). Substrate type significantly affected (P < 0.05) the insecticide knockdown response of the susceptible strain in acetone-based insecticide bioassays, with longer survival time on filter paper than on the glass surface. With the exception of deltamethrin, the different diluents (oil and acetone) also significantly affected (P < 0.05) the insecticide knockdown response of the susceptible strain in the filter paper-based insecticide bioassays, with longer survival time with acetone as the diluent. For both strains of C. hemipterus, there were no significant effects with the different surfaces and diluents for all insecticides except for malathion and imidacloprid, which was largely due to high levels of resistance. The lower effectiveness for the insecticide acetone-based treatment on filter paper may be due to crystal bloom. This occurs when an insecticide, dissolved in a volatile solvent, is applied onto absorptive surfaces. The effect is reduced on nonabsorptive surfaces and slowed down with oil-based insecticides, whereby the oil forms a film on absorptive surfaces. These findings suggest that nonabsorptive surfaces should be used in bioassays to monitor insecticide resistance. If absorptive surfaces are used in bioassays for testing active ingredients, then oil-based insecticides should be preferably used. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  5. Study on Biological Degradation of Industrial Organic Waste Residue with surface Soil%土壤有机废渣的生物降解研究

    Institute of Scientific and Technical Information of China (English)

    齐向阳; 周波

    2011-01-01

    [ Objective ] The study aimed to discuss the influencing factors and effective measures for the biological degradation of chemicalindustrial waste residue with the soil microbial. [ Method ] In testing area the soil samples were taken from the surface soil in 20cm with quartering method and the burning weightlessness of unit quality was detected resp., and then the different waste residue was applied in the each experimental plot and the their burning weightlessness were determined after taking the samples in the interval of 5 d, thus the degradation data of organic waste residue in each block of soil was acquired. [ Result ] The biological degradation of organic waste residue with the soil microbial was effected by the waste residue property and surface area, soil oxygenation content, soil pH, soil moisture content and soil temperatures. The aerobic degradation of organic matter was much faster and fuller than the anaerobic degradation. As the soil pH affected the microbial activities, it should be maintained at 7~9. Controlling the soil moisture content of 50%~60% was the best condition of microbial activity. When the soil temperature was below zero, the biological degradation stopped basically. [ Conclusion ] In the actual application of industrially processing the organic waste residue, the some soil texture and some kinds of wasted residue still needed for further research so as to control the biodegradation rate and degree and its management measures. texture.%[目的]探讨土壤微生物降解化工废渣的影响因素及有效措施.[方法]在试验区用四分法在地表20cm内取土壤样品,分别测出单位质量的燃烧失重,再向每块试验区施入不同的废渣,每隔5d取样后,测定其燃烧失重,获得各块土壤有机废渣随时间的降解数据.[结果]土壤有机废渣的生物降解受废渣性质、废渣表面积、土壤含氧量、土壤pH、含湿量和土壤温度的影响.有机物的

  6. Investigation of the occurrence of pesticide residues in rural wells and surface water following application to tobacco

    Directory of Open Access Journals (Sweden)

    Edson C. Bortoluzzi

    2007-01-01

    Full Text Available In this work the exposure of wells and surface water to pesticides, commonly used for tobacco cropping, was assessed. Water consumption wells and surface water flows were sampled at different times. After a preconcentration step with solid phase extraction (SPE, the selected pesticides were determined by gas chromatography with electron capture detection (GC-ECD or high performance liquid chromatography with diode array detection (HPLC-DAD. No pesticides were detected in the well water samples and surface water flow in the winter season. However, in the spring and summer higher concentrations of chlorpyrifos and imidacloprid were found in the water source samples. Atrazine, simazine and clomazone were also found. The occurrence of pesticides in collected water samples was related with the application to tobacco.

  7. Comparing ion conductance recordings of synthetic lipid bilayers with cell membranes containing TRP channels

    CERN Document Server

    Laub, Katrine R; Blicher, Andreas; Madsen, Soren B; Luckhoff, Andreas; Heimburg, Thomas

    2011-01-01

    In this article we compare electrical conductance events from single channel recordings of three TRP channel proteins (TRPA1, TRPM2 and TRPM8) expressed in human embryonic kidney cells with channel events recorded on synthetic lipid membranes close to melting transitions. Ion channels from the TRP family are involved in a variety of sensory processes including thermo- and mechano-reception. Synthetic lipid membranes close to phase transitions display channel-like events that respond to stimuli related to changes in intensive thermodynamic variables such as pressure and temperature. TRP channel activity is characterized by typical patterns of current events dependent on the type of protein expressed. Synthetic lipid bilayers show a wide spectrum of electrical phenomena that are considered typical for the activity of protein ion channels. We find unitary currents, burst behavior, flickering, multistep-conductances, and spikes behavior in both preparations. Moreover, we report conductances and lifetimes for lipi...

  8. A thermodynamic framework for understanding temperature sensing by transient receptor potential (TRP) channels.

    Science.gov (United States)

    Clapham, David E; Miller, Christopher

    2011-12-06

    The exceptionally high temperature sensitivity of certain transient receptor potential (TRP) family ion channels is the molecular basis of hot and cold sensation in sensory neurons. The laws of thermodynamics dictate that opening of these specialized TRP channels must involve an unusually large conformational standard-state enthalpy, ΔH(o): positive ΔH(o) for heat-activated and negative ΔH(o) for cold-activated TRPs. However, the molecular source of such high-enthalpy changes has eluded neurobiologists and biophysicists. Here we offer a general, unifying mechanism for both hot and cold activation that recalls long-appreciated principles of protein folding. We suggest that TRP channel gating is accompanied by large changes in molar heat capacity, ΔC(P). This postulate, along with the laws of thermodynamics and independent of mechanistic detail, leads to the conclusion that hot- and cold-sensing TRPs operate by identical conformational changes.

  9. Uterine deletion of Trp53 compromises antioxidant responses in mouse decidua

    Energy Technology Data Exchange (ETDEWEB)

    Burnum, Kristin E.; Hirota, Yasushi; Baker, Erin Shammel; Yoshie, Mikihiro; Ibrahim, Yehia M.; Monroe, Matthew E.; Anderson, Gordon A.; Smith, Richard D.; Daikoku, Takiko; Dey, Sudhansu K.

    2012-09-01

    Preterm birth is a global health issue impacting both mothers and children. However, the etiology of preterm birth is not clearly understood. From our recent finding that premature decidual senescence with terminal differentiation is a cause of preterm birth in mice with uterine Trp53 deletion, encoding p53 protein, led us to explore other potential factors that are related to preterm birth. Utilizing proteomics approaches, here we show that 183 candidate proteins cause significant changes in decidua with Trp53 deletion as compared to normal decidua. Functional categorization of these proteins unveiled new pathways that are influenced by p53. In particular, downregulation of a cluster of antioxidant proteins in p53 deficient decidua suggests that increased oxidative stress could be one cause of preterm birth in mice with uterine deletion of Trp53.

  10. Residues of organochlorine pesticides in surface soil and raw foods from rural areas of the Republic of Tajikistan

    Science.gov (United States)

    The central Asian Republic of Tajikistan has been an area of extensive historical agricultural pesticide use as well as large scale burials of obsolete banned chlorinated insecticides. The current investigation was a four year study of legacy organochlorine pesticides in surface ...

  11. Evidence for Novel Pharmacological Sensitivities of Transient Receptor Potential (TRP Channels in Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Swarna Bais

    2015-12-01

    Full Text Available Schistosomiasis, caused by parasitic flatworms of the genus Schistosoma, is a neglected tropical disease affecting hundreds of millions globally. Praziquantel (PZQ, the only drug currently available for treatment and control, is largely ineffective against juvenile worms, and reports of PZQ resistance lend added urgency to the need for development of new therapeutics. Ion channels, which underlie electrical excitability in cells, are validated targets for many current anthelmintics. Transient receptor potential (TRP channels are a large family of non-selective cation channels. TRP channels play key roles in sensory transduction and other critical functions, yet the properties of these channels have remained essentially unexplored in parasitic helminths. TRP channels fall into several (7-8 subfamilies, including TRPA and TRPV. Though schistosomes contain genes predicted to encode representatives of most of the TRP channel subfamilies, they do not appear to have genes for any TRPV channels. Nonetheless, we find that the TRPV1-selective activators capsaicin and resiniferatoxin (RTX induce dramatic hyperactivity in adult worms; capsaicin also increases motility in schistosomula. SB 366719, a highly-selective TRPV1 antagonist, blocks the capsaicin-induced hyperactivity in adults. Mammalian TRPA1 is not activated by capsaicin, yet knockdown of the single predicted TRPA1-like gene (SmTRPA in S. mansoni effectively abolishes capsaicin-induced responses in adult worms, suggesting that SmTRPA is required for capsaicin sensitivity in these parasites. Based on these results, we hypothesize that some schistosome TRP channels have novel pharmacological sensitivities that can be targeted to disrupt normal parasite neuromuscular function. These results also have implications for understanding the phylogeny of metazoan TRP channels and may help identify novel targets for new or repurposed therapeutics.

  12. Residue determination and levels of glyphosate in surface waters, sediments and soils associated with oil palm plantation in Tasik Chini, Pahang, Malaysia

    Science.gov (United States)

    Mardiana-Jansar, K.; Ismail, B. S.

    2014-09-01

    Levels of glyphosate and its main metabolite were determined in surface water, soil and sediment samples from an oil palm plantation area located at Tasik Chini, Pahang, Malaysia. The optimization analytical method has been developed for the determination of glyphosate herbicide and its metabolite amino-methyl-phosphonic acid (AMPA) in surface waters to a level of 0.1μg/L, while in sediments and soils to a level of 0.5μg/g with a good linearity in the calibration range of 1-100μg/L. The procedure involves a pre-columnderivatization step with 9-fluorenyl-methyl-chloroformate (FMOC-Cl) yielding highly fluorescent derivatives of the analytes which can be determined by HPLC with fluorescence detection. In the field, levels of glyphosate in surface waters ranges from not detected to 1.0mg/L, while in soils and sediments were from not detected to 6.0mg/kg. For AMPA, the residues in surface waters were between not detected to 2.0mg/L, while in soil and sediment samples were from not detected to 5mg/kg. This variation of glyphosate and AMPA levels depended directly on time of pesticide application and the season.

  13. Electrostatic contribution of surface charge residues to the stability of a thermophilic protein: benchmarking experimental and predicted pKa values.

    Directory of Open Access Journals (Sweden)

    Chi-Ho Chan

    Full Text Available Optimization of the surface charges is a promising strategy for increasing thermostability of proteins. Electrostatic contribution of ionizable groups to the protein stability can be estimated from the differences between the pKa values in the folded and unfolded states of a protein. Using this pKa-shift approach, we experimentally measured the electrostatic contribution of all aspartate and glutamate residues to the stability of a thermophilic ribosomal protein L30e from Thermococcus celer. The pKa values in the unfolded state were found to be similar to model compound pKas. The pKa values in both the folded and unfolded states obtained at 298 and 333 K were similar, suggesting that electrostatic contribution of ionizable groups to the protein stability were insensitive to temperature changes. The experimental pKa values for the L30e protein in the folded state were used as a benchmark to test the robustness of pKa prediction by various computational methods such as H++, MCCE, MEAD, pKD, PropKa, and UHBD. Although the predicted pKa values were affected by crystal contacts that may alter the side-chain conformation of surface charged residues, most computational methods performed well, with correlation coefficients between experimental and calculated pKa values ranging from 0.49 to 0.91 (p<0.01. The changes in protein stability derived from the experimental pKa-shift approach correlate well (r = 0.81 with those obtained from stability measurements of charge-to-alanine substituted variants of the L30e protein. Our results demonstrate that the knowledge of the pKa values in the folded state provides sufficient rationale for the redesign of protein surface charges leading to improved protein stability.

  14. Down-regulation of tyrosinase, TRP-1, TRP-2 and MITF expressions by citrus press-cakes in murine B16 F10 melanoma

    Institute of Scientific and Technical Information of China (English)

    Sang Suk Kim; Min-Jin Kim; Young Hun Choi; Byung Kuk Kim; Kwang Sik Kim; Kyung Jin Park; Suk Man Park; Nam Ho Lee; Chang-Gu Hyun

    2013-01-01

    Objective: To investigate the suitability of citrus-press cakes, by-products of the juice industry as a source for the whitening agents for cosmetic industry. Methods:Ethylacetate extracts of citrus-press cakes (CCE) were examined for their anti-melanogenic potentials in terms of the inhibition of melanin production and mechanisim of melanogenesis by using Western Blot analysis with tyrosinese, tyrosinase-related protein-1 (TRP-1), TRP2, and microphthalmia-associated transcription factor (MITF) proteins. To apply the topical agents, citrus-press cakes was investigated the safety in human skin cell line. Finally flavonoid analysis of CCE was also determined by HPLC analysis. Results: Results indicated that CCE were shown to down-regulate melanin content in a dose-dependent pattern. The CCE inhibited tyrosinase, TRP-2, and MITF expressions in a dose-dependent manner. To test the applicability of CCE to human skin, we used MTT assay to assess the cytotoxic effects of CCE on human keratinocyte HaCaT cells. The CCE exhibited low cytotoxicity at 50 µg/mL. Characterization of the citrus-press cakes for flavonoid contents using HPLC showed varied quantity of rutin, narirutin, and hesperidin. Conclusions:Considering the anti-melanogenic activity and human safety, CCE is considered as a potential anti-melanogenic agent and may be effective for topical application for treating hyperpigmentation disorders.

  15. Ehrlichia chaffeensis TRP32 is a Nucleomodulin that Directly Regulates Expression of Host Genes Governing Differentiation and Proliferation.

    Science.gov (United States)

    Farris, Tierra R; Dunphy, Paige S; Zhu, Bing; Kibler, Clayton E; McBride, Jere W

    2016-08-29

    Ehrlichia chaffeensis is an obligately intracellular bacterium that reprograms the mononuclear phagocyte through diverse effector-host interactions to modulate numerous host cell processes, including transcription. In a previous study, we reported that E. chaffeensis TRP32, a type 1 secreted effector, interacts with multiple host nucleus-associated proteins and also auto-activates reporter gene expression in yeast. In this study, we demonstrate that TRP32 is a nucleomodulin that binds host DNA and alters host gene transcription. TRP32 enters the host cell nucleus via a noncanonical translocation mechanism that involves phosphorylation of Y179 located in a C-terminal tri-tyrosine motif. Both genistein and mutation of Y179 inhibited TRP32 nuclear entry. An electromobility shift assay (EMSA) demonstrated TRP32 host DNA binding via its tandem repeat domain. TRP32 DNA binding and motif preference were further confirmed by supershift assays, as well as competition and mutant probe analyses. Using ChIP-Seq, we determined that TRP32 binds a G-rich motif primarily within ±500 bp of the gene transcription start site. An ontology analysis identified genes involved in processes such as immune cell differentiation, chromatin remodeling, and RNA transcription and processing, as primary TRP32 targets. TRP32 bound genes (n=1223) were distributed on all chromosomes and included several global regulators of proliferation and inflammation such as FOS and JUN, AKT3 and NRAS, and non-coding RNA genes, miRNA 21 and miRNA 142. TRP32 target genes were differentially regulated during infection, the majority of which were repressed, and direct repression/activation of these genes by TRP32 was confirmed in vitro with a cellular luciferase reporter assay.

  16. Mutations in the exon 7 of Trp53 gene and the level of p53 protein in double transgenic mouse model of Alzheimer's disease.

    Science.gov (United States)

    Dorszewska, Jolanta; Oczkowska, Anna; Suwalska, Monika; Rozycka, Agata; Florczak-Wyspianska, Jolanta; Dezor, Mateusz; Lianeri, Margarita; Jagodzinski, Paweł P; Kowalczyk, Michal J; Prendecki, Michal; Kozubski, Wojciech

    2014-01-01

    Alzheimer's disease (AD) leads to generation of β-amyloid (Aβ) in the brain. Alzheimer's disease model PS/APP mice show a markedly accelerated accumulation of Aβ, which may lead to apoptosis induction e.g. in cells expressing wild-type p53. The TP53 gene is found to be the most frequently mutated gene in human tumour cells. There is accumulating evidence pointing out to the contribution of oxidative stress and chronic inflammation in both AD and cancer. The purpose of this study was to analyze exon 7 mutations of the murine Trp53 gene and Aβ/A4 and p53 protein levels in PS/APP and control mice. The studies were performed on female double transgenic PS/APP mice and young adults (8-12 weeks old) and age-matched control mice. The Trp53 mutation analysis was carried out with the use of PCR and DNA sequencing. The Aβ/A4 and p53 levels were analyzed by Western blotting. The frequency of mutations was almost quadrupled in PS/APP mice (44%), compared to controls (14%). PS/APP mice with the A929T and A857G mutations had a similar p53 level. In cerebral gray matter of PS/APP mice the level of p53 positive correlated with the level of Aβ protein (RS = +0.700, p p53 down-regulation, while in aging ones, G859A substitution was most likely associated with over-expression of p53. In silico protein analysis revealed a possibly substantial impact of all four mutations on p53 activity. Three mutations were in close proximity to zinc-coordinating cysteine residues. It seems that in PS/APP mice missense Trp53 exon 7 mutations may be associated with the degenerative process by changes of p53 protein function.

  17. Kinetic network study of the diversity and temperature dependence of Trp-Cage folding pathways: combining transition path theory with stochastic simulations.

    Science.gov (United States)

    Zheng, Weihua; Gallicchio, Emilio; Deng, Nanjie; Andrec, Michael; Levy, Ronald M

    2011-02-17

    We present a new approach to study a multitude of folding pathways and different folding mechanisms for the 20-residue mini-protein Trp-Cage using the combined power of replica exchange molecular dynamics (REMD) simulations for conformational sampling, transition path theory (TPT) for constructing folding pathways, and stochastic simulations for sampling the pathways in a high dimensional structure space. REMD simulations of Trp-Cage with 16 replicas at temperatures between 270 and 566 K are carried out with an all-atom force field (OPLSAA) and an implicit solvent model (AGBNP). The conformations sampled from all temperatures are collected. They form a discretized state space that can be used to model the folding process. The equilibrium population for each state at a target temperature can be calculated using the weighted-histogram-analysis method (WHAM). By connecting states with similar structures and creating edges satisfying detailed balance conditions, we construct a kinetic network that preserves the equilibrium population distribution of the state space. After defining the folded and unfolded macrostates, committor probabilities (P(fold)) are calculated by solving a set of linear equations for each node in the network and pathways are extracted together with their fluxes using the TPT algorithm. By clustering the pathways into folding "tubes", a more physically meaningful picture of the diversity of folding routes emerges. Stochastic simulations are carried out on the network, and a procedure is developed to project sampled trajectories onto the folding tubes. The fluxes through the folding tubes calculated from the stochastic trajectories are in good agreement with the corresponding values obtained from the TPT analysis. The temperature dependence of the ensemble of Trp-Cage folding pathways is investigated. Above the folding temperature, a large number of diverse folding pathways with comparable fluxes flood the energy landscape. At low temperature

  18. Atomically flat surface of (0 0 1) textured FePt thin films by residual stress control

    Science.gov (United States)

    Liu, S. H.; Hsiao, S. N.; Chou, C. L.; Chen, S. K.; Lee, H. Y.

    2015-11-01

    Single-layered Fe52Pt48 films with thickness of 10 nm were sputter-deposited on glass substrates. Rapid thermal annealing with different heating rates (10-110 K/s) was applied to transform as-deposited fcc phase into L10 phase and meanwhile to align [0 0 1]-axis of L10 crystal along plane normal direction. Based on X-ray diffractometry using synchrotron radiation source, the texture coefficient of (0 0 1)-plane increases with increasing heating rate from 10 to 40 K/s, which is correlated with perpendicular magnetic anisotropy and in-plane tensile stress analyzed by asymmetric sin2 ψ method. Furthermore, it was revealed by atomic force microscopy that the dewetting process occurred as heating rate was raised up to 80 K/s and higher. The change in the microstructure due to stress relaxation leads to the degradation of (0 0 1) orientation and magnetic properties. Surface roughness is closely related to the in-plane tensile stress. Enhanced perpendicular magnetic anisotropy and atomically flat surface were achieved for the samples annealed at 40 K/s, which may be suitable for further practical applications. This work also suggests a feasible way for surface engineering by controlling internal stress of the FePt without introducing cap layer.

  19. Using Standing Gold Nanorod Arrays as Surface-Enhanced Raman Spectroscopy (SERS) Substrates for Detection of Carbaryl Residues in Fruit Juice and Milk.

    Science.gov (United States)

    Alsammarraie, Fouad K; Lin, Mengshi

    2017-01-25

    In recent years, there have been increasing concerns about pesticide residues in various foods. On the other hand, there is growing attention in utilizing novel nanomaterials as highly sensitive, low-cost, and reproducible substrates for surface-enhanced Raman spectroscopy (SERS) applications. The objective of this study was to develop a SERS method for the rapid detection of pesticides that were extracted from different types of food samples (fruit juice and milk). A new SERS substrate was prepared by assembling gold nanorods into standing arrays on a gold-coated silicon slide. The standing nanorod arrays were neatly arranged and were able to generate a strong electromagnetic field in SERS measurement. The as-prepared SERS substrate was utilized to detect carbaryl in acetonitrile/water solution, fruit juices (orange and grapefruit), and milk. The results show that the concentrations of carbaryl spiked in fruit juice and milk were linearly correlated with the concentrations predicted by the partial least-squares (PLS) models with r values of 0.91, 0.88, and 0.95 for orange juice, grapefruit juice, and milk, respectively. The SERS method was able to detect carbaryl that was extracted from fruit juice and milk samples at a 50 ppb level. The detection limits of carbaryl were 509, 617, and 391 ppb in orange juice, grapefruit juice, and milk, respectively. All detection limits are below the maximum residue limits that were set by the U.S. EPA. Moreover, satisfactory recoveries (82-97.5%) were accomplished for food samples using this method. These results demonstrate that SERS coupled with the standing gold nanorod array substrates is a rapid, reliable, sensitive, and reproducible method for the detection of pesticide residues in foods.

  20. Research on the influence of machining introduced sub-surface defects and residue stress upon the mechanical properties of single crystal copper

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Large scale molecular dynamics simulations of nanomachining and stretching of single crystal copper are performed to analyze the machining process’ influence on the material’s mechanical properties. The simulation results show that the machining process will introduce interfacial defects inside the specimen and enhance the compressive stress beneath the surface. Gener- ally speaking, interfacial defects lead to the decrease of the strength limit, while residue compressive stress can enhance the elastic limit and even the strength limit. Various machining parameters are adopted to investigate their influence on the me- chanical behavior of machined specimen. Lower cutting speed and smaller cutting depth lead to less defects and greater residue compressive stress, which brings about better mechanical properties. The elastic limit increases by 36.8% under the cutting depth of 0.73 nm and decreases by 21.1% under the cutting depth of 1.46 nm. The strength limit increases by 7.7% under the cutting speed of 100 m/s and decreases by 28.2% under the cutting speed of 300 m/s.

  1. Residuation theory

    CERN Document Server

    Blyth, T S; Sneddon, I N; Stark, M

    1972-01-01

    Residuation Theory aims to contribute to literature in the field of ordered algebraic structures, especially on the subject of residual mappings. The book is divided into three chapters. Chapter 1 focuses on ordered sets; directed sets; semilattices; lattices; and complete lattices. Chapter 2 tackles Baer rings; Baer semigroups; Foulis semigroups; residual mappings; the notion of involution; and Boolean algebras. Chapter 3 covers residuated groupoids and semigroups; group homomorphic and isotone homomorphic Boolean images of ordered semigroups; Dubreil-Jacotin and Brouwer semigroups; and loli

  2. Examination of Surface Residuals Obtained During Re-Lubrication of the International Space Station (ISS) Solar Alpha Rotary Joint (SARJ)

    Science.gov (United States)

    Martinez, J. E.; Golden, J. L.

    2012-01-01

    The starboard SARJ mechanism on the ISS suffered a premature lubrication failure, resulting in widespread loss of the nitride case layer on its 10.3 meter circumference, 15-5PH steel race ring [1, 2]. To restore functionality, vacuum-stable grease was applied on-orbit, first to the port SARJ mechanism to save it from the damage suffered by the starboard mechanism. After 3 years of greased operation, telemetry indicated that the port mechanism required relubrication, so part of that process included sampling each of the three race ring surfaces to evaluate any wear debris recovered and the state of the originally applied grease. Extensive microscopic examination was conducted, which directed subsequent microanalysis of particulate. Since the SARJ mechanism operates in the vacuum of space, a sampling method and tool had to be developed for use by astronauts while working in the extravehicular mobility unit (EMU). The sampling tool developed was a cotton terry-cloth mitt for the EMU glove, with samples taken by swiping each of the three port SARJ race-ring surfaces. The sample mitts for each surface were folded inward after sampling to preserve sample integrity, for return and ground analysis. The sample mitt for what is termed the outer canted surface of the SARJ race-ring is shown in Figure 1. Figure 1 also demonstrates how increasing levels of magnification were used to survey the contamination removed in sampling, specifically looking for signs of wear debris or other features which could be further evaluated using Scanning Electron Microscopy (SEM) methods. The most surprising overall result at this point in the analysis was the relatively small amounts of grease recovered during sampling. It is clear that the mechanism was not operating with surplus lubricant. Obviously, evidence of molybdenum disulfide (MoS2), a major component in the grease applied, was prevalent in the analysis conducted. But a small amount of mechanism wear debris was observed. Figure 2

  3. Nuclear and perinuclear targeting efficiency of quantum dots depends on density of peptidic targeting residues on their surface.

    Science.gov (United States)

    Maity, Amit Ranjan; Stepensky, David

    2016-12-29

    Targeted delivery to the cell nucleus can enhance the efficiency of drugs with nuclear site of action (some anti-cancer agents, DNA drugs, etc.), and can reduce their toxicity. Such targeting can be attained using nano-drug delivery systems (nano-DDSs) decorated with nuclear targeting sequences (such as nuclear localization sequence peptides, NLS). Several types of nano-DDSs decorated with NLS peptides were designed, but their investigation usually did not include quantitate analysis of the decoration efficiency and its correlation with the nano-DDSs intracellular localization. Thus, the major mechanisms and limiting factors of the nano-DDSs nuclear targeting are largely unknown yet. In this study, we report quantitative data for specific nano-formulation (CdSe-ZnS quantum dots) that include the efficiencies of its decoration with NLS residues and of its nuclear and perinuclear targeting, and demonstrate correlation between these parameters. For instance, QDs decorated with 83, 246, and 265 NLS peptides accumulated efficiently in the nucleus of HeLa cells or its vicinity (an average of 30.4%, 43.3%, and 49.0% of the intracellular QDs, respectively). On the other hand, QDs decorated with 63, 231, and 308 scrambled peptides accumulated in the nucleus of HeLa cells or its vicinity to a much lower extent (an average of 17.3%, 21.1%, and 25.5% of the intracellular QDs, respectively). Thus, results of our study provide important insights into the structure-activity correlations (i.e., the relationships between the formulation properties and the intracellular fate of nano-DDSs) of nuclear-targeted drug delivery. We plan to apply the research tools that were developed in the course of this and our previous studies to investigate the nuclear and perinuclear targeting activities of different NLS sequences, and to investigate the effects of nano-DDSs size, charge, shape, decoration efficiency with nuclear targeting sequences, and other structural factors on nuclear and

  4. Residual stresses in a surface remelting of castings made of cobalt alloy MAR-M509 with a plasma generated in electric arc

    Directory of Open Access Journals (Sweden)

    Z. Opiekun

    2010-01-01

    Full Text Available The manuscript presents the results of measurements of residual stresses (RS in partial meltings of casting surfaces made of cobalt alloy MAR-M509. The partial meltings were made with an argon plasma beam by GTAW method. The values of RS were deter-mined by X-ray diffraction method in grazing incident geometry, by g-sin2ψ method and in Bragg-Brentano (BB geometry. It has been stated that RS values depend on the parameters of partial melting process. It has been claimed that compressive stresses, which are present in the thin layer up to ca 2 μm, convert to tensile stresses in deeper layers of partial meltings.

  5. Process Optimization of Ultrasonic-Assisted Extraction of Arabinogalactan from Dihydroquercetin Extracted Residues by Response Surface Methodology and Evaluation of Its Antioxidant Activity

    Directory of Open Access Journals (Sweden)

    Zaizhi Liu

    2013-01-01

    Full Text Available Ultrasound was used for the extraction of larch arabinogalactan from Larix gmelinii. The optimal conditions for ultrasound extraction were determined by response surface methodology. Specifically, the Box-Behnken design was employed to evaluate the effects of three independent variables: ultrasound time, temperature, and liquid-solid ratio. The highest arabinogalactan yield (11.18% was obtained under the optimal extraction condition (extraction temperature 41.5°C, extraction time 24.3 min, and liquid-solid ratio 40 mL/g. In addition, the antioxidant activity of arabinogalactan that was extracted from dihydroquercetin extraction residues exhibited a moderate and concentration-dependent hydroxyl radical-scavenging capacity, ferric-reducing power, and reducing power. The wood material was characterized before and after processing by scanning electron microscopy and Fourier-transform infrared spectroscopy.

  6. Elevated expression of TrpC5 and GLUT1 is associated with chemoresistance in colorectal cancer.

    Science.gov (United States)

    Wang, Teng; Ning, Kuan; Lu, Ting-Xun; Hua, Dong

    2017-02-01

    Reprogramming of energy metabolism (aerobic glycolysis) is thought to play an essential role in cancer. Compared to oxidative phosphorylation, aerobic glycolysis consumes more glucose through the upregulation of glucose transporters, notably glucose transporter 1 (GLUT1). Elevated glycolysis occurs in chemoresistant cancer cells, but the detailed mechanism is not well understood. The upregulation of the Ca2+-permeable transient receptor potential channel 5 (TrpC5) activates the Wnt/β-catenin signaling pathway in 5-fluorouracil (5-Fu)-resistant human colorectal cancer (CRC) HCT-8 (HCT-8/5-Fu) cells. In the present study, TrpC5 was overexpressed at the mRNA and protein levels along with GLUT1 in HCT-8/5-Fu cells. Suppression of TrpC5 expression with a TrpC5-specific shRNA reduced the induction of GLUT1 in the HCT-8 cells. The inhibition of the Wnt/β-catenin signaling pathway with XAV939 resulted in a decreased GLUT1 and nuclear c-Myc expression. Further study using clinical specimens validated the positive correlation between TrpC5 and GLUT1 protein levels and showed that a high TrpC5/GLUT1 expression was significantly correlated with chemoresistance. Taken together, we demonstrated the essential role of TrpC5 in GLUT1 induction and revealed that a high TrpC5/GLUT1 expression is associated with chemoresistance in human CRC.

  7. A C. elegans stretch receptor neuron revealed by a mechanosensitive TRP channel homologue.

    Science.gov (United States)

    Li, Wei; Feng, Zhaoyang; Sternberg, Paul W; Xu, X Z Shawn

    2006-03-30

    The nematode Caenorhabditis elegans is commonly used as a genetic model organism for dissecting integration of the sensory and motor systems. Despite extensive genetic and behavioural analyses that have led to the identification of many genes and neural circuits involved in regulating C. elegans locomotion behaviour, it remains unclear whether and how somatosensory feedback modulates motor output during locomotion. In particular, no stretch receptors have been identified in C. elegans, raising the issue of whether stretch-receptor-mediated proprioception is used by C. elegans to regulate its locomotion behaviour. Here we have characterized TRP-4, the C. elegans homologue of the mechanosensitive TRPN channel. We show that trp-4 mutant worms bend their body abnormally, exhibiting a body posture distinct from that of wild-type worms during locomotion, suggesting that TRP-4 is involved in stretch-receptor-mediated proprioception. We show that TRP-4 acts in a single neuron, DVA, to mediate its function in proprioception, and that the activity of DVA can be stimulated by body stretch. DVA both positively and negatively modulates locomotion, providing a unique mechanism whereby a single neuron can fine-tune motor activity. Thus, DVA represents a stretch receptor neuron that regulates sensory-motor integration during C. elegans locomotion.

  8. Prostaglandin H synthase-mediated bioactivation of the amino acid pyrolysate product Trp P-2

    Energy Technology Data Exchange (ETDEWEB)

    Petry, T.W.; Krauss, R.S.; Eling, T.E.

    1986-08-01

    We report evidence that the mutagen and carcinogen 3-amino-1-methyl-5H pyrido(4,3b)indole (Trp P-2) is a substrate for co-oxidation by prostaglandin H synthase (PHS) in ram seminal vesicle (RSV) microsomes. Trp P-2 serves as a reducing cofactor for the hydroperoxidase activity of PHS as shown by the concentration-dependent inhibition of the hydroperoxidase catalyzed incorporation of molecular oxygen into phenylbutazone. Spectral data suggest that this metabolism results in disruption of the double bond conjugation within the nucleus of the molecule. A single metabolite peak which was dependent upon arachidonic acid and substrate concentration was separated from the parent compound by h.p.l.c. following incubation with RSV microsomes. Co-oxidation of Trp P-2 produced reactive intermediates which bound covalently to microsomal protein (9 nmol/mg) and to calf thymus DNA (475 pmol/mg). Binding was inhibited by indomethacin, and supported by substitution of hydrogen peroxide for arachidonic acid. These data suggest a possible role for PHS in the in situ activation of Trp P-2 to its ultimate carcinogenic form in tissues which contain PHS.

  9. Rate constant and reaction coordinate of Trp-cage folding in explicit water

    NARCIS (Netherlands)

    Juraszek, J.; Bolhuis, P.G.

    2008-01-01

    We report rate constant calculations and a reaction coordinate analysis of the rate-limiting folding and unfolding process of the Trp-cage mini-protein in explicit solvent using transition interface sampling. Previous transition path sampling simulations revealed that in this (un)folding process the

  10. Efficient and reproducible folding simulations of the Trp-case protein with multiscale molecular dynamics

    Institute of Scientific and Technical Information of China (English)

    XIA XueFeng; ZHANG Song; HUANG go; ZHOU Yun; SUN ZhiRong

    2008-01-01

    Folding simulations are often time-consuming or highly sensitive to the initial conformation of the simulation even for mini protein like the Trp-cage. Here, we present a multiscale molecular dynamics method which appears to be both efficient and insensitive to the starting conformation based on the testing results from the Trp-cage protein. In this method the simulated system is simultaneously modeled on atoms and coarse-grained particles with incremental coarsening levels. The dynamics of coarse-grained particles are adapted to the recent trajectories of finer-grained particles instead of fixed and parameterized energy functions as used in previous coarse-grained models. In addition, the compositions of coarse-grained particles are allowed to be updated automatically based on the coherence during its history. Starting from the fully extended conformation and other several different conformations of the Trp-cage protein, our method successfully finds out the native-like conformations of the Trp-cage protein in the largest cluster of the trajectories in all of the eight performed simulations within at most 10 ns simulation time. The results show that approaches based on multiscale modeling are promising for ab initio protein structure prediction.

  11. Emerging role of TRP channels in cell migration: from tumor vascularization to metastasis

    Directory of Open Access Journals (Sweden)

    Alessandra eFioro Pla

    2013-11-01

    Full Text Available Transient Receptor Potential (TRP channels modulate intracellular Ca2+ concentrations, controlling critical cytosolic and nuclear events that are involved in the initiation and progression of cancer. It is not, therefore, surprising that the expression of some TRP channels is altered during tumor growth and metastasis. Cell migration of both epithelial and endothelial cells is an essential step of the so-called metastatic cascade that leads to the spread of the disease within the body. It is in fact required for both tumor vascularization as well as for tumor cell invasion into adjacent tissues and intravasation into blood/lymphatic vessels. Studies from the last 15 years have unequivocally shown that the ion channles and the transport proteins also play important roles in cell migration. On the other hand, recent literature underlies a critical role for TRP channels in the migration process both in cancer cells as well as in tumor vascularization. This will be the main focus of our review. We will provide an overview of recent advances in this field describing TRP channels contribution to the process vascular and cancer cell migration, and we will systematically discuss relevant molecular mechanism involved.

  12. Twin-screw extruded lipid implants containing TRP2 peptide for tumour therapy.

    Science.gov (United States)

    Even, Marie-Paule; Bobbala, Sharan; Gibson, Blake; Hook, Sarah; Winter, Gerhard; Engert, Julia

    2017-01-16

    Much effort has been put in the development of specific anti-tumour immunotherapies over the last few years, and several studies report on the use of liposomal carriers for tumour-associated antigens. In this work, the use of lipid implants, prepared using two different extruders, was investigated for sustained delivery in tumour therapy. The implants consisted of cholesterol, soybean lecithin, Dynasan 114, trehalose, ovalbumin (OVA) or a TRP2 peptide, and Quil-A. Implants were first produced on a Haake Minilab extruder, and then a scale-down to minimal quantities of material on a small scale ZE mini extruder was performed. All formulations were characterised in terms of extrudability, implant properties and in vitro release behaviour of the model antigen ovalbumin. The type of extruder used to produce the implants had a major influence on implant properties and the release behaviour, demonstrating that extrusion parameters and lipid formulations have to be individually adapted to each extrusion device. Subsequently, lipid implants containing TRP-2 peptide were extruded on the ZE mini extruder and investigated in vitro and in vivo. The in vivo study showed that mice having received TRP2 loaded implants had delayed tumour growth for 3days compared to groups having received no TRP2.

  13. Alpha-lactalbumin combined with a regular diet increases plasma Trp-LNAA ratio

    NARCIS (Netherlands)

    Beulens, J.W.J.; Bindels, J.G.; Graaf, de C.

    2004-01-01

    Brain serotonin influences food intake and mood. It is synthesised from tryptophan (Tip) of which uptake in the brain is dependent on plasma ratio of tryptophan to the sum of other large neutral amino acids (Trp-LNAA). A carbohydrate-rich diet increases this ratio, whereas a protein-rich diet decrea

  14. Residual activity of methoprene and novaluron as surface treatments to manage the flour beetles, Tribolium castaneum and Tribolium confusum.

    Science.gov (United States)

    Arthur, Frank H; Fontenot, Emily A

    2012-01-01

    The juvenile hormone analog methoprene, and the chitin synthesis inhibitor novaluron, were evaluated by exposing late-stage larvae of Tribolium castaneum (Herbst) or Tribolium confusum (Jacqueline DuVal) (Coleoptera: Tenebrionidae) to it. The larvae were exposed to it in food material, on concrete, on plywood, and on floor tile. Larvae of T. castaneum were more susceptible than T. confusum larvae to both methoprene and novaluron on all surfaces. A further evaluation was done by exposing adult T. confusum to methoprene and novaluron through food placed on concrete treated with methoprene and novaluron, and then assessing resulting progeny production. The emergence of adults with normal morphology was reduced for both chemicals, with more malformed adults appearing in the methoprene treatment, and fewer adults of any form emerging in the novaluron treatment. The results show direct exposures to larvae, or determining progeny production from exposed adults, are valid methods for assessing the susceptibility of flour beetles to insecticides.

  15. Numerical analysis of residual stress of Al-Mg-Mn-Sc-Zr alloy subjected to surface strengthening by shot peening

    Directory of Open Access Journals (Sweden)

    Mariusz Stegliński

    2015-03-01

    Full Text Available In this paper, we presented the results of the analysis of the stresses in the Al-Mg5%-Mn1,5%-Sc0,8%-Zr0,4% alloy after shot peening process using solver ANSYSANSYSANSYS LS-Dyna. The computational model illustrates the phenomena occurring as a result of plastic deformation caused by hitting a steel ball on the surface of the analyzed aluminium alloy. We analyzed two input variables: diameter and speed of a ball. The resulting normal stress distribution centred exposes the minimum compressive stress at a position located at a depth point of Belayev 0.125 mm with a value of σ = –345 MPa. Variable parameter shows the correlation of the boundary conditions of minimum stress increase with increasing ball’s diameter and its speed. Selected points of numerical analysis were verified with experimental results.[b]Keywords[/b]: materials science, numerical analysis, metal forming, shot peening, aluminium

  16. [Impact of polymetallic mine (Zn, Pb, Cu) residues on surface water, sediments and soils at the vicinity (Marrakech, Morocco)].

    Science.gov (United States)

    El Adnani, M; Rodriguez-Maroto, J M; Sbai, M L; Loukili Idrissi, L; Nejmeddine, A

    2007-09-01

    Metal sulphide tailings present a potential risk for the environment because of their natural oxidability which leads to the production of acid mine drainage. The prospected site close to Marrakech includes zinc, lead and copper sulphide deposits. This site is located in an agricultural area near the Tensift River which is used for irrigation. In addition to the tailing leachates, underground mine waters are also discharged into the river. This represents a potential risk for the environment and human health. The aim of this study was to assess the tailings impact on surface water, sediments and soil qualities. Chemical analysis of surface water and sediments collected downstream of the mine revealed that, water and sediments present high concentrations of major ions and heavy metals. The analysis also revealed spatial as well as temporal changes in the chemical properties of the studied water and sediments. These changes are attributed to the rising phenomena. The soil near the mine presents high content of sulphate. Its Zn, Pb, Cu and Fe contents are respectively 38, 15, 11 and 1.6 times higher than non contaminated soils located far away from the site. The soil irrigated with underground mine waters shows concentrations of SO4(2-), Cu, Zn, Fe, Cd and Pb which are respectively 4, 10, 28, 2, 9 and 12 times higher than soils which are not irrigated with this mine water. This study also showed that there has been a change in the physicochemical characteristics of water and sediments in the sampling points downstream of the mine before its closure and after its activity renewal.

  17. Is Asp-His-Ser/Thr-Trp tetrad hydrogen-bond network important to WD40-repeat proteins: a statistical and theoretical study.

    Science.gov (United States)

    Wu, Xian-Hui; Zhang, Hui; Wu, Yun-Dong

    2010-04-01

    WD40-repeat proteins are abundant and play important roles in forming protein complexes. The domain usually has seven WD40 repeats, which folds into a seven beta-sheet propeller with each beta-sheet in a four-strand structure. An analysis of 20 available WD40-repeat proteins in Protein Data Bank reveals that each protein has at least one Asp-His-Ser/Thr-Trp (D-H-S/T-W) hydrogen-bonded tetrad, and some proteins have up to six or seven such tetrads. The relative positions of the four residues in the tetrads are also found to be conserved. A sequence alignment analysis of 560 WD40-repeat protein sequences in human reveals very similar features, indicating that such tetrad may be a general feature of WD40-repeat proteins. We carried out density functional theory and found that these tetrads can lead to significant stabilization including hydrogen-bonding cooperativity. The hydrogen bond involving Trp is significant. These results lead us to propose that the tetrads may be critical to the stability and the mechanism of folding of these proteins.

  18. A Trp474Cys mutation in the alpha-subunit of beta-hexosaminidase causes a subacute encephalopathic form of G{sub M2} gangliosidosis, type 1

    Energy Technology Data Exchange (ETDEWEB)

    Petroulakis, E.; Cao, Z.; Salo, T. [Univ. of Manitoba, Winnipeg (Canada)] [and others

    1994-09-01

    Mutations in the HEXA gene that encodes the {alpha}-subunit of the heterodimeric lysosomal enzyme {beta}-hexosaminidase A, or Hex A ({alpha}{beta}), cause G{sub M2} gangliosidosis, type 1. The infantile form (Tay-Sachs disease) results when there is no residual Hex A activity, while less severe and more variable clinical phenotypes result when residual Hex A activity is present. A non-Jewish male who presented with an acute psychotic episode at age 16 was diagnosed with a subacute encephalopathic form of G{sub M2} gangliosidosis. At age 19, chronic psychosis with intermittent acute exacerbations remains the most disabling symptom in this patient and his affected brother although both exhibit some ataxia and moderately severe dysarthria. We have found a 4 bp insertion (+TATC 1278) associated with infantile Tay-Sachs disease on one allele; no previously identified mutation was found on the second allele. SSCP analysis detected a shift in exon 13 and sequencing revealed a G1422C mutation in the second allele that results in a Trp474Cys substitution. The presence of the mutation was confirmed by the loss of HaeIII and ScrFI sites in exon 13 PCR products from the subjects and their father. The mutation was introduced into the {alpha}-subunit cDNA and Hex S ({alpha}{alpha}) and Hex A ({alpha}{beta}) were transiently expressed in monkey COS-7 cells. The Trp474Cys mutant protein had approximately 5% and 12% of wild-type Hex S and Hex A activity, respectively. Western blot analysis revealed a small amount of residual mature {alpha}-subunit and a normal level of precursor protein. We conclude that the Trp474Cys mutation is the cause of the Hex A deficiency associated with a subacute (juvenile-onset) phenotype in this patient. Like other mutations in exon 13 of HEXA, it appears to affect intracellular processing. Studies of the defect in intracellular processing are in progress.

  19. Transient receptor potential (TRP) channels as drug targets for diseases of the digestive system.

    Science.gov (United States)

    Holzer, Peter

    2011-07-01

    Approximately 20 of the 30 mammalian transient receptor potential (TRP) channel subunits are expressed by specific neurons and cells within the alimentary canal. They subserve important roles in taste, chemesthesis, mechanosensation, pain and hyperalgesia and contribute to the regulation of gastrointestinal motility, absorptive and secretory processes, blood flow, and mucosal homeostasis. In a cellular perspective, TRP channels operate either as primary detectors of chemical and physical stimuli, as secondary transducers of ionotropic or metabotropic receptors, or as ion transport channels. The polymodal sensory function of TRPA1, TRPM5, TRPM8, TRPP2, TRPV1, TRPV3 and TRPV4 enables the digestive system to survey its physical and chemical environment, which is relevant to all processes of digestion. TRPV5 and TRPV6 as well as TRPM6 and TRPM7 contribute to the absorption of Ca²⁺ and Mg²⁺, respectively. TRPM7 participates in intestinal pacemaker activity, and TRPC4 transduces muscarinic acetylcholine receptor activation to smooth muscle contraction. Changes in TRP channel expression or function are associated with a variety of diseases/disorders of the digestive system, notably gastro-esophageal reflux disease, inflammatory bowel disease, pain and hyperalgesia in heartburn, functional dyspepsia and irritable bowel syndrome, cholera, hypomagnesemia with secondary hypocalcemia, infantile hypertrophic pyloric stenosis, esophageal, gastrointestinal and pancreatic cancer, and polycystic liver disease. These implications identify TRP channels as promising drug targets for the management of a number of gastrointestinal pathologies. As a result, major efforts are put into the development of selective TRP channel agonists and antagonists and the assessment of their therapeutic potential.

  20. Impact of mining residues on surface and groundwater quality. Case of the mining sector of Azzaba in the North-East of Algeria

    Science.gov (United States)

    Haouli, Zouina; Kherici, Nacer; Derdous, Oussama; Sassane, Amina; Bougherira, Nabil

    2016-07-01

    Azzaba region contains a mining sector that was created in 1971 and operated until 2006, during this period huge quantities of mining residues were released in nature without any environmental rehabilitation plan which certainly deteriorated surface and groundwater quality by trace metals menacing the people's health as well as the aquatic ecosystems in this zone. The purpose of this study is to illustrate and to assess the surface and groundwater pollution toward heavy metals at the vicinity of the abandoned mining site. The primary analysis aimed to evaluate the pollution due to mercury in the region after many years of the closure of the mining industry to compare it with evaluations made during its operational period. In addition, further analyses of water pollution toward heavy metals usually used in the mercury industry (iron, zinc and copper) and probably released in the Fendek Wadi were conducted. These analyses allowed characterizing the ecological state of the studied environment by highlighting the concentrations of trace elements (mercury, iron, zinc, copper). According to the analyses, most of these concentrations meet the World Health Organization (WHO) standards; in fact only iron concentration exceeds them at the stations P7 and P8. Finally, the study results were compared by those obtained by previous studies; it was found that the mercury concentration has decreased with time which means that the contamination danger is disappearing.

  1. Residual-stress measurements

    Energy Technology Data Exchange (ETDEWEB)

    Ezeilo, A.N.; Webster, G.A. [Imperial College, London (United Kingdom); Webster, P.J. [Salford Univ. (United Kingdom)

    1997-04-01

    Because neutrons can penetrate distances of up to 50 mm in most engineering materials, this makes them unique for establishing residual-stress distributions non-destructively. D1A is particularly suited for through-surface measurements as it does not suffer from instrumental surface aberrations commonly found on multidetector instruments, while D20 is best for fast internal-strain scanning. Two examples for residual-stress measurements in a shot-peened material, and in a weld are presented to demonstrate the attractive features of both instruments. (author).

  2. Microenvironment of tryptophan residues in beta-lactoglobulin derivative polypeptide-sodium dodecyl sulfate complexes.

    Science.gov (United States)

    Imamura, T; Konishi, K

    1992-06-01

    The changes of microenvironment of tryptophan residues in beta-lactoglobulin A and its cyanogen bromide (CNBr) fragments with the binding of sodium dodecyl sulfate (SDS) were studied with measurements of the rates of N-bromosuccinimide (NBS) modification reactions by stopped-flow photometry. Two tryptophan residues of carboxyamidomethylated (RCM) beta-lactoglobulin A in the states of their complexes with SDS were clearly distinguishable by their differences in NBS modification rates. We confirmed by experiments with CNBr fragments containing trytophan residue. The modification rates of Trp 19 in RCM beta-lactoglobulin A-SDS complexes were about 10-fold smaller than those expected for tryptophan residues exposed entirely to the aqueous solvent. The Trp 61 was hardly changed. The change of rate constants for Trp 19 was virtually consistent with those observed when N-acetyl-L-trytophan ethylester was dissolved in SDS micelles. For various species of polypeptide-SDS complexes, all tryptophan residues were reactive to NBS and also, for some of them, the differences in NBS modification rates were observed between tryptophan residues on a common polypeptide chain. These results suggest micellar and heterogeneous bindings of SDS to polypeptides.

  3. 老采空区地表残余沉降预计与应用%Prediction of Surface Residual Settlement Over the Old Goal

    Institute of Scientific and Technical Information of China (English)

    王磊; 郭广礼; 查剑锋; 张鲜妮; 栗帅; 徐斗斗

    2011-01-01

    为了对长壁垮落法开采老采空区"活化"引起的地表残余沉降进行估计,从老采空区的覆岩形态结构出发,提出把长壁垮落法开采老采空区分成2个区进行预计:边界的欠压密区和中部的压密区,给出了欠压密区和压密区的尺寸.针对边界"活化"空间的分布特征和突发性"活化"特点,提出等价成变采高的小工作面开采来预计边界"活化"引起的地表残余沉降.针对中部破碎岩体缓慢蠕变的"活化"特点,采用剩余下沉系数进行预计.分别建立了相应的预计模型,并编写了预计程序.工程实践表明,老采空区的边界蕴含着大量"活化"空间,若边界处结构失稳,将在其正上方地表产生较大的不均匀移动变形;在采宽不大的情况下,老采空区"活化"引起的最大残余沉降位置在开切眼和停采线附近上方地表,并非在采空区中部压密区.%To estimate surface residual settlement caused by “activation” of old goal with longwall caving, we propose, for the first time, that the old goal is divided into two areas based on the morphological structure of overlying strata. These two areas are: 1) the border browndense area, and 2) the central compaction zone. In addition, we have given the size of browndense area and the compaction zone. Aiming at the spatial distribution characteristics of border “activation” and the sudden happening of border “activation” , we propose the new method that a small working face with a variable mining height is used to predict surface residual settlement caused by boder “activation”. In view of the slow creep “activation” feature of central broken rock, the remaining subsidence coefficient is used for the prediction. Further more, we have established the prediction model and compiled the prediction program. Finally, we have got the conclusion that the old goal boder area contains a large number of “activated” spaces. If the boundary structure becomes

  4. Time-resolved EPR studies with DNA photolyase: excited-state FADH0 abstracts an electron from Trp-306 to generate FADH-, the catalytically active form of the cofactor.

    Science.gov (United States)

    Kim, S T; Sancar, A; Essenmacher, C; Babcock, G T

    1993-09-01

    Photolyase repairs UV-induced cyclobutane-pyrimidine dimers in DNA by photoinduced electron transfer. The enzyme isolated from Escherichia coli contains 5,10-methenyltetrahydrofolate, which functions as the light-harvesting chromophore, and fully reduced flavin adenine dinucleotide (FAD), which functions as the redox catalyst. During enzyme preparation, the flavin is oxidized to FADH0, which is catalytically inert. Illumination of the enzyme with 300- to 600-nm light converts the flavin to the fully reduced form in a reaction that involves photooxidation of an amino acid in the apoenzyme. The results of earlier optical studies had indicated that the redox-active amino acid in this photoactivation process was tryptophan. We have now used time-resolved electron paramagnetic resonance (EPR) spectroscopy to investigate the photoactivation reaction. Excitation of the flavin-radical-containing inactive enzyme produces a spin-polarized radical that we identify by 2H and 15N labeling as originating from a tryptophan residue, confirming the inferences from the optical work. These results and Trp-->Phe replacement by site-directed mutagenesis reveal that flavin radical photoreduction is achieved by electron abstraction from Trp-306 by the excited-state FADH0. Analysis of the hyperfine couplings and spin density distribution deduced from the isotopic-labeling results shows that the product of the light-driven redox chemistry is the Trp-306 cation radical. The results strongly suggest that the active form of photolyase contains FADH- and not FADH2.

  5. A competing hydrophobic tug on L596 to the membrane core unlatches S4-S5 linker elbow from TRP helix and allows TRPV4 channel to open.

    Science.gov (United States)

    Teng, Jinfeng; Loukin, Stephen H; Anishkin, Andriy; Kung, Ching

    2016-10-18

    We have some generalized physical understanding of how ion channels interact with surrounding lipids but few detailed descriptions on how interactions of particular amino acids with contacting lipids may regulate gating. Here we discovered a structure-specific interaction between an amino acid and inner-leaflet lipid that governs the gating transformations of TRPV4 (transient receptor potential vanilloid type 4). Many cation channels use a S4-S5 linker to transmit stimuli to the gate. At the start of TRPV4's linker helix is leucine 596. A hydrogen bond between the indole of W733 of the TRP helix and the backbone oxygen of L596 secures the helix/linker contact, which acts as a latch maintaining channel closure. The modeled side chain of L596 interacts with the inner lipid leaflet near the polar-nonpolar interface in our model-an interaction that we explored by mutagenesis. We examined the outward currents of TRPV4-expressing Xenopus oocyte upon depolarizations as well as phenotypes of expressing yeast cells. Making this residue less hydrophobic (L596A/G/W/Q/K) reduces open probability [Po; loss-of-function (LOF)], likely due to altered interactions at the polar-nonpolar interface. L596I raises Po [gain-of-function (GOF)], apparently by placing its methyl group further inward and receiving stronger water repulsion. Molecular dynamics simulations showed that the distance between the levels of α-carbons of H-bonded residues L596 and W733 is shortened in the LOFs and lengthened in the GOFs, strengthening or weakening the linker/TRP helix latch, respectively. These results highlight that L596 lipid attraction counteracts the latch bond in a tug-of-war to tune the Po of TRPV4.

  6. Assignment of ozone-sensitive tryptophan residue in tryptophanase by a dual-monitoring high-performance liquid chromatography system

    Energy Technology Data Exchange (ETDEWEB)

    Ida, N.; Tokushige, M.

    1985-02-01

    Tryptophanase purified from Escherichia coli B/1t7-A is inactivated by mild ozonization following pseudo-first-order kinetics. Previous data from the authors suggest that one out of two tryptophan residues (Trp's) in the enzyme subunit is preferentially oxidized concomitant with the ozone inactivation and has a direct interaction with the coenzyme, pyridoxal phosphate. To determine which Trp is more susceptible to ozonization and interacts with PLP, the native and ozonized enzyme proteins were cleaved by trypsin and the two Trp-containing peptides were analyzed by reverse-phase HPLC equipped with a dual-monitoring system consisting of an uv and a fluorescence monitor connected in tandem for selective detection of Trp-containing peptides. This device facilitated rapid detection and quantitation of the Trp-containing peptides which decreased upon ozonization. The results showed that Trp preferentially oxidized upon ozonization and involved in the interaction with PLP was the one in peptide T-15 rather than that in T-23, which Kagamiyama et al. originally designated.

  7. Primary structural response in tryptophan residues of Anabaena sensory rhodopsin to photochromic reactions of the retinal chromophore

    Science.gov (United States)

    Inada, Seisuke; Mizuno, Misao; Kato, Yoshitaka; Kawanabe, Akira; Kandori, Hideki; Wei, Zhengrong; Takeuchi, Satoshi; Tahara, Tahei; Mizutani, Yasuhisa

    2013-06-01

    Anabaena sensory rhodopsin (ASR) is a microbial rhodopsin found in eubacteria and functions as a photosensor. The photoreaction of ASR is photochromic between all-trans, 15-anti (ASRAT), and 13-cis, 15-syn (ASR13C) isomers. To understand primary protein dynamics in the photoreaction starting in ASRAT and ASR13C, picosecond time-resolved ultraviolet resonance Raman spectra were obtained. In the intermediate state appearing in the picosecond temporal region, spectral changes of Trp bands were observed. For both ASRAT and ASR13C, the intensities of the Trp bands were bleached within the instrumental response time and recovered with a time constant of 30 ps. This suggests that the rates of structural changes in the Trp residue in the vicinity of the chromophore do not depend on the direction of the isomerization of retinal. A comparison between spectra of the wild-type and Trp mutants indicates that the structures of Trp76 and Trp46 change upon the primary photoreaction of retinal.

  8. Molecular basis of epithelial Ca2+ and Mg2+ transport: insights from the TRP channel family

    DEFF Research Database (Denmark)

    Dimke, Henrik Anthony; Hoenderop, Joost G J; Bindels, René J M

    2011-01-01

    potential (TRP) family identified several new ion channels acting as gatekeepers of Ca(2+) and Mg(2+) transport in these epithelia, greatly increasing our understanding of the molecular processes that facilitate the movement of these minerals. In the intestine, TRP channels contribute to the saturable...... active transcellular movement of divalent cations from the lumen into the enterocyte. Furthermore, in bone, TRPV channels play important roles by influencing the osteoclastic resorption process, thereby contributing importantly to overall bone mineral content. The divalent cation-permeable TRPV5 and TRPM......6 channels are located in the renal distal convolution, the main site of active transcellular Ca(2+) and Mg(2+) transport. The channels are regulated by a multitude of factors and hormones that contribute importantly to keeping the systemic concentrations of Ca(2+) and Mg(2+) within normal limits...

  9. Structure and function of Escherichia coli met repressor: similarities and contrasts with trp repressor.

    Science.gov (United States)

    Phillips, S E; Stockley, P G

    1996-04-29

    Transcription of genes encoding enzymes for the biosynthesis of methionine and trytophan in Escherichia coli is regulated by the ligand-activated met and trp repressors. X-ray crystallographic studies show how these two small proteins, although similar in size and function, have totally different three-dimensional structures and specifically recognize their respective DNA operator sequences in different ways. A common feature is that both repressors bind as cooperative arrays to tandem repeats of 8 base-pair 'Met' or 'Trp boxes' respectively, and the consensus sequences share the rare tetranucleotide CTAG. A series of structural and functional studies have shown how the two repressors discriminate between their operators, using a combination of direct contacts between side chains and bases, and indirect sensing of conformational properties of the DNA.

  10. A TRP Channel in the Lysosome Regulates Large Particle Phagocytosis via Focal Exocytosis

    OpenAIRE

    2013-01-01

    Phagocytosis of large extracellular particles such as apoptotic bodies requires delivery of the intracellular endosomal and lysosomal membranes to form plasmalemmal pseudopods. Here we identified Mucolipin TRP channel 1 (TRPML1) as the key lysosomal Ca2+ channel regulating focal exocytosis and phagosome biogenesis. Both particle ingestion and lysosomal exocytosis are inhibited by synthetic TRPML1 blockers, and are defective in macrophages isolated from TRPML1 knockout mice. Furthermore, TRPML...

  11. Crystal Structures of Mite Allergens Der f 1 and Der p 1 Reveal Differences in Surface-Exposed Residues that May Influence Antibody Binding

    Energy Technology Data Exchange (ETDEWEB)

    Chruszcz, Maksymilian; Chapman, Martin D.; Vailes, Lisa D.; Stura, Enrico A.; Saint-Remy, Jean-Marie; Minor, Wladek; Pomés, Anna; (INDOOR Bio.); (CEAS); (Leuven); (UV)

    2009-12-01

    The Group 1 mite allergens, Der f 1 and Der p 1, are potent allergens excreted by Dermatophagoides farinae and Dermatophagoides pteronyssinus, respectively. The human IgE antibody responses to the Group 1 allergens show more cross-reactivity than the murine IgG antibody responses which are largely species-specific. Here, we report the crystal structure of the mature form of Der f 1, which was isolated from its natural source, and a new, high-resolution structure of mature recombinant Der p 1. Unlike Der p 1, Der f 1 is monomeric both in the crystalline state and in solution. Moreover, no metal binding is observed in the structure of Der f 1, despite the fact that all amino acids involved in Ca{sup 2+} binding in Der p 1 are completely conserved in Der f 1. Although Der p 1 and Der f 1 share extensive sequence identity, comparison of the crystal structures of both allergens revealed structural features which could explain the differences in murine and human IgE antibody responses to these allergens. There are structural differences between Der f 1 and Der p 1 which are unevenly distributed on the allergens' surfaces. This uneven spatial arrangement of conserved versus altered residues could explain both the specificity and cross-reactivity of antibodies against Der f 1 and Der p 1.

  12. Experimental and Numerical Investigation of the Effect of Process Conditions on Residual Wall Thickness and Cooling and Surface Characteristics of Water-Assisted Injection Molded Hollow Products

    Directory of Open Access Journals (Sweden)

    Hyungpil Park

    2015-01-01

    Full Text Available Recently, water-assisted injection molding was employed in the automobile industry to manufacture three-dimensional hollow tube-type products with functionalities. However, process optimization is difficult in the case of water-assisted injection molding because of the various rheological interactions between the injected water and the polymer. In this study, the boiling phenomenon that occurs because of the high melt temperature when injecting water and the molding characteristics of the hollow section during the water-assisted injection process were analyzed by a water-assisted injection molding analysis. In addition, the changes in the residual wall thickness accompanying changes in the process conditions were compared with the analysis results by considering water-assisted injection molding based on gas-assisted injection molding. Furthermore, by comparing the cooling characteristics and inner wall surface qualities corresponding to the formation of the hollow section by gas and water injections, a water-assisted injection molding technique was proposed for manufacturing hollow products with functionality.

  13. Efficacy and duration of three residual insecticides on cotton duck and vinyl tent surfaces for control of the sand fly Phlebotomus papatasi (Diptera: Psychodidae).

    Science.gov (United States)

    Zayed, Abdel Baset B; Hoel, David F; Tageldin, Reham A; Fawaz, Emaldeldin Y; Furman, Barry D; Hogsette, Jerome A; Bernier, Ulrich R

    2013-01-01

    This study evaluated the toxicity and duration of 3 residual insecticides against the Old World sand fly, Phlebotomus papatasi, an important vector of cutaneous leishmaniasis, on 2 types of tent material used by the US military in Afghanistan and the Middle East. Vinyl and cotton duck tent surfaces were treated at maximum labeled rates of lambda-cyhalothrin (Demand CS, Zeneca Inc, Wilmington, DE), bifenthrin (Talstar P Professional, FMC Corporation, Philadelphia, PA) and permethrin (Insect Repellent, Clothing Application, 40%), then subsequently stored in indoor, shaded spaces at room temperature (60%-70% relative humidity (RH), 22°C-25°C), and under sunlight and ambient air temperatures outdoors (20%-30% RH, 29°C-44°C). Insecticide susceptible colony flies (F110) obtained from the insectary of US Naval Medical Research Unit No. 3, Cairo, Egypt, were exposed to treated tent surfaces for 30-minute periods twice monthly for up to 5 months, then once monthly thereafter, using the World Health Organization cone assay. Lambda-cyhalothrin treated cotton duck tent material stored indoors killed P. papatasi for 8 months, while the complementary sun-exposed cotton duck material killed adult flies for 1 month before the efficacy dropped to less than 80%. Sand fly mortality on permethrin- and bifenthrin-treated cotton duck decreased below 80% after 2 weeks exposure to sunlight. Shade-stored permethrin and bifenthrin cotton duck material killed more than 80% of test flies through 5 months before mortality rates decreased substantially. Vinyl tent material provided limited control (less than 50% mortality) for less than 1 month with all treatment and storage regimes. Lambda-cyhalothrin-treated cotton duck tent material provided the longest control and produced the highest overall mortalities (100% for 8 months (shaded), more than 90% for 1 month (sunlight-exposed)) of both cotton duck and vinyl tents.

  14. Synthesis of Z-CCK-27-32-NH2, Z-Tyr(SO-3)-Met-Gly-Trp-Met-Asp-NH2, a cholecystokinin receptor antagonist and an inhibitor of gastrin-induced acid secretion.

    Science.gov (United States)

    Briet, C; Aumelas, A; Martinez, J

    1985-09-01

    The synthesis of the hexapeptide Z-Tyr(SO-3)-Met-Gly-Trp-Met-Asp-NH2, representing the C-terminal sequence of cholecystokinin minus the C-terminal phenylalanyl residue is described. This peptide was shown to be the most potent cholecystokinin receptor antagonist in vitro described to date. It is also able to inhibit gastrin-induced acid secretion in vivo, in the rat and was proved to antagonize the action of the C-terminal octapeptide of cholecystokinin in the central nervous system.

  15. TrpC5 Mediates Acute Leptin and Serotonin Effects via Pomc Neurons

    Directory of Open Access Journals (Sweden)

    Yong Gao

    2017-01-01

    Full Text Available The molecular mechanisms underlying acute leptin and serotonin 2C receptor-induced hypophagia remain unclear. Here, we show that neuronal and pro-opiomelanocortin (Pomc-specific loss of transient receptor potential cation 5 (TrpC5 subunits is sufficient to decrease energy expenditure and increase food intake resulting in elevated body weight. Deficiency of Trpc5 subunits in Pomc neurons is also sufficient to block the anorexigenic effects of leptin and serotonin 2C receptor (Ht2Cr agonists. The loss of acute anorexigenic effects of these receptors is concomitant with a blunted electrophysiological response to both leptin and Ht2Cr agonists in arcuate Pomc neurons. We also demonstrate that the Ht2Cr agonist lorcaserin-induced improvements in glucose and insulin tolerance are blocked by TrpC5 deficiency in Pomc neurons. Together, our results link TrpC5 subunits in the brain with leptin- and serotonin 2C receptor-dependent changes in neuronal activity, as well as energy balance, feeding behavior, and glucose metabolism.

  16. ZBTB20 regulates nociception and pain sensation by modulating TRP channel expression in nociceptive sensory neurons.

    Science.gov (United States)

    Ren, An-Jing; Wang, Kai; Zhang, Huan; Liu, Anjun; Ma, Xianhua; Liang, Qing; Cao, Dongmei; Wood, John N; He, David Z; Ding, Yu-Qiang; Yuan, Wen-Jun; Xie, Zhifang; Zhang, Weiping J

    2014-11-05

    In mammals, pain sensation is initiated by the detection of noxious stimuli through specialized transduction ion channels and receptors in nociceptive sensory neurons. Transient receptor potential (TRP) channels are the key sensory transducers that confer nociceptors distinct sensory modalities. However, the regulatory mechanisms about their expression are poorly defined. Here we show that the zinc-finger protein ZBTB20 regulates TRP channels expression in nociceptors. ZBTB20 is highly expressed in nociceptive sensory neurons of dorsal root ganglia. Disruption of ZBTB20 in nociceptors led to a marked decrease in the expression levels of TRPV1, TRPA1 and TRPM8 and the response of calcium flux and whole-cell currents evoked by their respective specific agonists. Phenotypically, the mice lacking ZBTB20 specifically in nociceptors showed a defect in nociception and pain sensation in response to thermal, mechanical and inflammatory stimulation. Our findings point to ZBTB20 as a critical regulator of nociception and pain sensation by modulating TRP channels expression in nociceptors.

  17. Inhibition of autophagy attenuates pancreatic cancer growth independent of TP53/TRP53 status.

    Science.gov (United States)

    Yang, Annan; Kimmelman, Alec C

    2014-09-01

    Basal levels of autophagy are elevated in most pancreatic ductal adenocarcinomas (PDAC). Suppressing autophagy pharmacologically using chloroquine (CQ) or genetically with RNAi to essential autophagy genes inhibits human pancreatic cancer growth in vitro and in vivo, which presents possible treatment opportunities for PDAC patients using the CQ-derivative hydroxychloroquine (HCQ). Indeed, such clinical trials are ongoing. However, autophagy is a complex cellular mechanism to maintain cell homeostasis under stress. Based on its biological role, a dual role of autophagy in tumorigenesis has been proposed: at tumor initiation, autophagy helps maintain genomic stability and prevent tumor initiation; while in advanced disease, autophagy degrades and recycles cellular components to meet the metabolic needs for rapid growth. This model was proven to be the case in mouse lung tumor models. However, in contrast to prior work in various PDAC model systems, loss of autophagy in PDAC mouse models with embryonic homozygous Trp53 deletion does not inhibit tumor growth and paradoxically increases progression. This raised concerns whether there may be a genotype-dependent reliance of PDAC on autophagy. In a recent study, our group used a Trp53 heterozygous mouse PDAC model and human PDX xenografts to address the question. Our results demonstrate that autophagy inhibition was effective against PDAC tumors irrespective of TP53/TRP53 status.

  18. Pulse radiolysis studies of intramolecular electron transfer in model peptides and proteins. 7. Trp -> TyrO radical transformation in hen egg-white lysozyme. Effects of pH, temperature, Trp62 oxidation and inhibitor binding

    DEFF Research Database (Denmark)

    Bobrowski, K.; Holcman, J.; Poznanski, J.

    1997-01-01

    (.). Arrhenius plots of the temperature-dependence of k(5) showed that the activation energy of LRET varies both with temperature and the protonation state of the enzyme. The activation energies are in the range 7.6-56.0 kJ mol(-1) and are similar to those for activation of amide hydrogen exchange in native HEWL...... below its denaturation temperature. Selective oxidation by ozone of the Trp62 indole side-chain in HEWL to N'-formylkynurenine (NFKyn62-HEWL) caused a large drop in the initial yield of Trp(.) radicals, G(Trp(.))(i). This was accompanied by a relatively small decrease in k(5) but selective oxidation...... by ozone had a pronounced effect on its temperature-dependence. Taken together these observations indicate that of the six tryptophans present in HEWL Trp62 contributes about 50% to the yield of the observed LRET. In the enzyme-inhibitor complex, HEWL(GlcNAc)(3), where Trp62 and Trp63 are completely...

  19. TRP channels, omega-3 fatty acids, and oxidative stress in neurodegeneration: from the cell membrane to intracellular cross-links

    Directory of Open Access Journals (Sweden)

    M. Leonelli

    2011-11-01

    Full Text Available The transient receptor potential channels family (TRP channels is a relatively new group of cation channels that modulate a large range of physiological mechanisms. In the nervous system, the functions of TRP channels have been associated with thermosensation, pain transduction, neurotransmitter release, and redox signaling, among others. However, they have also been extensively correlated with the pathogenesis of several innate and acquired diseases. On the other hand, the omega-3 polyunsaturated fatty acids (n-3 fatty acids have also been associated with several processes that seem to counterbalance or to contribute to the function of several TRPs. In this short review, we discuss some of the remarkable new findings in this field. We also review the possible roles played by n-3 fatty acids in cell signaling that can both control or be controlled by TRP channels in neurodegenerative processes, as well as both the direct and indirect actions of n-3 fatty acids on TRP channels.

  20. The role of hydrophobic active-site residues in substrate specificity and acyl transfer activity of penicillin acylase

    NARCIS (Netherlands)

    Alkema, WBL; Dijkhuis, AJ; de Vries, E; Janssen, DB

    2002-01-01

    Penicillin acylase of Escherichia colt catalyses the hydrolysis and synthesis of beta-lactam antibiotics. To study the role of hydrophobic residues in these reactions, we have mutated three active-site phenylalanines. Mutation of alphaF146, betaF24 and betaF57 to Tyr, Trp, Ala or Leu yielded mutants

  1. A cation-pi interaction in the binding site of the glycine receptor is mediated by a phenylalanine residue

    DEFF Research Database (Denmark)

    Pless, Stephan Alexander; Millen, Kat S; Hanek, Ariele P;

    2008-01-01

    Cys-loop receptor binding sites characteristically contain many aromatic amino acids. In nicotinic ACh and 5-HT3 receptors, a Trp residue forms a cation-pi interaction with the agonist, whereas in GABA(A) receptors, a Tyr performs this role. The glycine receptor binding site, however, contains pr...

  2. Host determinant residue lysine 627 lies on the surface of a discrete, folded domain of influenza virus polymerase PB2 subunit.

    Directory of Open Access Journals (Sweden)

    Franck Tarendeau

    Full Text Available Understanding how avian influenza viruses adapt to human hosts is critical for the monitoring and prevention of future pandemics. Host specificity is determined by multiple sites in different viral proteins, and mutation of only a limited number of these sites can lead to inter-species transmission. Several of these sites have been identified in the viral polymerase, the best characterised being position 627 in the PB2 subunit. Efficient viral replication at the relatively low temperature of the human respiratory tract requires lysine 627 rather than the glutamic acid variant found systematically in avian viruses. However, the molecular mechanism by which any of these host specific sites determine host range are unknown, although adaptation to host factors is frequently evoked. We used ESPRIT, a library screening method, to identify a new PB2 domain that contains a high density of putative host specific sites, including residue 627. The X-ray structure of this domain (denoted the 627-domain exhibits a novel fold with the side-chain of Lys627 solvent exposed. The structure of the K627E mutated domain shows no structural differences but the charge reversal disrupts a striking basic patch on the domain surface. Five other recently proposed host determining sites of PB2 are also located on the 627-domain surface. The structure of the complete C-terminal region of PB2 comprising the 627-domain and the previously identified NLS-domain, which binds the host nuclear import factor importin alpha, was also determined. The two domains are found to pack together with a largely hydrophilic interface. These data enable a three-dimensional mapping of approximately half of PB2 sites implicated in cross-species transfer onto a single structural unit. Their surface location is consistent with roles in interactions with other viral proteins or host factors. The identification and structural characterization of these well-defined PB2 domains will help design

  3. Construction of a Trp- commercial baker's yeast strain by using food-safe-grade dominant drug resistance cassettes.

    Science.gov (United States)

    Estruch, Francisco; Prieto, José Antonio

    2003-12-01

    We have designed a food-safe-grade module for gene disruptions in commercial baker's yeast strains, which contains the G418 resistance cassette, KanMX4, flanked by direct repeats from the MEL1 gene of Saccharomyces cerevisiae. This module was used to obtain a Trp(-) auxotrophic mutant of the polyploid HY strain by successive targeting to the TRP1 locus and later in vivo excision of the kan(r) marker. Southern blot analysis indicated that HY contains five copies of the TRP1 gene. However, after four disruption rounds, a strain named HYtrpM(4), unable to grow in the absence of tryptophan, was selected. Southern and Northern analysis of HYtrpM(4) cells showed that a remaining functional wild-type copy was still present, suggesting that the level of phosphoribosylanthranylate isomerase activity, resulting from a single copy of TRP1, is too low to sustain growth. Accordingly, a high reversion frequency of the Trp(-) phenotype, through gene conversion, was found in cells of the mutant strain. Nevertheless, this was not a drawback for its use as a recipient strain of heterologous genes. Indeed, YEpACT-X24 transformants were stable after 25 generations and expressed and secreted high levels of active recombinant xylanase. These data indicate that the new Trp(-) strain can be used to generate a stable recombinant yeast that fulfils all the requirements and market criteria for commercial utilisation.

  4. The cataract-associated V41M mutant of human γS-crystallin shows specific structural changes that directly enhance local surface hydrophobicity

    Energy Technology Data Exchange (ETDEWEB)

    Bharat, Somireddy Venkata; Shekhtman, Alexander; Pande, Jayanti, E-mail: jpande@albany.edu

    2014-01-03

    Highlights: •We present NMR analysis of V41M, a cataract-causing mutant of human γS-crystallin. •Mutation alters strand–strand interactions throughout the N-terminal domain. •Mutation directly affects Trp46 due to key Met41-S–Trp46-pi interactions. •We identify the basis of the surface hydrophobicity increase and residues involved. -- Abstract: The major crystallins expressed in the human lens are γS-, γC- and γD-crystallins. Several mutations in γS-crystallin are associated with hereditary cataracts, one of which involves the substitution of a highly conserved Valine at position 41 to Methionine. According to a recent report, the mutant protein, V41M, shows lower stability and increased surface hydrophobicity compared to the wild-type, and a propensity for self-aggregation. Here we address the structural differences between the two proteins, with residue-level specificity using NMR spectroscopy. Based on the structural model of the mutant protein, our results clearly show that the mutation creates a major local perturbation almost at the junction of the first and second “Greek-key” motifs in the N-terminal domain. A larger section of the second motif (residues 44–86) appears to be mainly affected. Based on the sizeable chemical shift of the imino proton of the indole side-chain of Trp46 in V41M, we suggest that the sulphur atom of Met41 is involved in an S–π interaction with Trp46. This interaction would bring the last β-strand of the first “Greek-key” motif closer to the first β-strand of the second motif. This appears to lead to a domino effect, towards both the N- and C-terminal ends, even as it decays off substantially beyond the domain interface. During this process discreet hydrophobic surface patches are created, as revealed by ANS-binding. Such changes would not affect the secondary structure or cause a major change in the tertiary structure, but can lead to self-aggregation or aberrant binding interactions of the mutant

  5. Cloning and Expression of trp Operon in Escherichia coli%大肠埃希菌trp operon的克隆与表达

    Institute of Scientific and Technical Information of China (English)

    林维平; 刘晓影; 武敬亮; 高志芹; 孙同毅

    2008-01-01

    色氨酸操纵子所表达酶的高效表达和酶活性的提高,从而构建高产色氨酸菌株.利用PCR的方法从大肠埃希菌基因组中直接克隆色氨酸操纵子,并将其连接到原核表达载体pBV220中,得到重组质粒pBV220-trp operon,转化大肠埃希菌DH5α,温度诱导重组蛋白表达,表达产物经SDS-PAGE分析并用比色法测定其活性.通过凝胶电泳观察PCR扩增产物大小约为7 kb.SDS-PAGE鉴定目的蛋白得到了高效表达,邻氨基苯甲酸合成酶和色氨酸合成酶的活性分别比对照提高了3.4倍和2.5倍.成功构建了重组质粒pBV220-trp operon,邻氨基苯甲酸合成酶和色氨酸合成酶的表达量和表达活性在大肠埃希菌中得到了提高,为高产色氨酸基因工程菌的构建奠定基础.

  6. 钛合金切削加工表面残余应力有限元仿真%Finite Element Simulation on Residual Stress of Machined Titanium Alloy Surface

    Institute of Scientific and Technical Information of China (English)

    刘文文; 刘长毅

    2012-01-01

    采用Johnson- Cook失效准则,建立了钛合金的二维正交切削热-机械应力耦合有限元仿真模型,分析计算了不同切削条件下已加工表面残余应力的分布规律.结果表明:已加工表面层残余应力为拉应力,沿着深度方向由拉应力逐渐过渡到压应力.表面残余应力随着切削速度的增大而增大,在一定的前角变化范围内,随着刀具前角的增大,表面残余拉应力先增大后减小,而随着刀具后角的增大却减小.各加工参数对残余应力层的厚度影响都很小.%Based on Johnson - Cook failure criterion, it establishes 2D orthogonal TC4 cutting model of the coupled temperature - mechanical stress, analyzes the distribution of surface residual stress under different cutting conditions. The results show that the surface residual stress of workpiece is tensile stress. Along the depth direction, the tensile stress is transformed to the compressive stress. Surface residual stress increases with the in-creasing of the cutting speed, in a certain range of rake angle, the tensile residual stress on the workpiece surface increases with rake angle at first and decreases subsequently, while decreases with the in - creasing of the relief angle. All cutting conditions have little influence on the thickness of the residual stress layer.

  7. Identification and characterization of TRP14, a thioredoxin-related protein of 14 kDa from orange-spotted grouper, Epinephelus coioides.

    Science.gov (United States)

    Wei, Jingguang; Ji, Huasong; Guo, Minglan; Yan, Yang; Qin, Qiwei

    2013-11-01

    Thioredoxin (abbreviated as Trx) is an important ubiquitous disulfide reductase, which can protect organisms against various oxidative stresses. In the present study, a thioredoxin-related protein of 14 kDa (named as Ec-TRP14) was identified from the marine fish grouper, Epinephelus coioides by RACE PCR. The full-length cDNA of Ec-TRP14 was comprised of 1066 bp with a 372 bp open reading frame that encodes a putative protein of 123 amino acids. Similar to most TRP14s, Ec-TRP14 contained the conserved motif C-P-D-C. Ec-TRP14 mRNA is predominately expressed in liver, brain and muscle. The expression of Ec-TRP14 was up-regulated in the liver of grouper challenged with SGIV. Ec-TRP14 was recombined and expressed in Escherichia coli BL21 (DE3), and the rEc-Ec-TRP14 fusion protein was demonstrated to possess the antioxidant activity. The grouper spleen (GS) cells were treated with a high concentration of rEc-TRP14 (8.3 μg/ml), which significantly enhanced cells viability under damage caused by viral infection. These results together indicated that Ec-TRP14 could function as an important antioxidant in a physiological context, and might be involved in the responses to viral challenge.

  8. Experimental determination of residual stress

    Science.gov (United States)

    Ferguson, Milton W.

    1991-01-01

    Residual stresses in finished parts have often been regarded as factors contributing to premature part failure and geometric distortions. Currently, residual stresses in welded structures and railroad components are being investigated. High residual stresses formed in welded structures due primarily to the differential contractions of the weld material as it cools and solidifies can have a profound effect on the surface performance of the structure. In railroad wheels, repeated use of the brakes causes high residual stresses in the rims which may lead to wheel failure and possible derailment. The goals of the study were: (1) to develop strategies for using x-ray diffraction to measure residual stress; (2) to subject samples of Inconel 718 to various mechanical and heat treatments and to measure the resulting stress using x-ray diffraction; and (3) to measure residual stresses in ferromagnetic alloys using magnetoacoustics.

  9. Identification of a quorum sensing pheromone posttranslationally farnesylated at the internal tryptophan residue from Bacillus subtilis subsp. natto.

    Science.gov (United States)

    Hayashi, Shunsuke; Usami, Syohei; Nakamura, Yuta; Ozaki, Koki; Okada, Masahiro

    2015-01-01

    Bacillus subtilis subsp. natto produces poly-γ-glutamic acid under the control of quorum sensing. We identified ComXnatto pheromone as the quorum-sensing pheromone with an amino acid sequence of Lys-Trp-Pro-Pro-Ile-Glu and the tryptophan residue posttranslationally modified by a farnesyl group. ComXnatto pheromone is unique in the sense that the 5th tryptophan residue from the C-terminal is farnesylated.

  10. Effect of texture and blasting pressure on residual stress and surface modifications in wet sand blasted α-Al2O3 coating

    OpenAIRE

    Ekström, Erik

    2015-01-01

    Recently, wet sand blasting on coated cutting tool inserts has drawn interest to the tooling industry due to its positive effects on cutting performance and tool life. This performance boost has partly been attributed to the buildup of compressive residual stresses in the coating during the blasting process. However, the mechanism of forming residual stresses in ceramic coatings during sand blasting is not fully understood. This work utilize x-ray diffraction as the main tool to study the for...

  11. Synergy between Prkdc and Trp53 regulates stem cell proliferation and GI-ARS after irradiation.

    Science.gov (United States)

    Gurley, Kay E; Ashley, Amanda K; Moser, Russell D; Kemp, Christopher J

    2017-11-01

    Ionizing radiation (IR) is one of the most widely used treatments for cancer. However, acute damage to the gastrointestinal tract or gastrointestinal acute radiation syndrome (GI-ARS) is a major dose-limiting side effect, and the mechanisms that underlie this remain unclear. Here we use mouse models to explore the relative roles of DNA repair, apoptosis, and cell cycle arrest in radiation response. IR induces DNA double strand breaks and DNA-PK mutant Prkdc(scid/scid) mice are sensitive to GI-ARS due to an inability to repair these breaks. IR also activates the tumor suppressor p53 to trigger apoptotic cell death within intestinal crypt cells and p53 deficient mice are resistant to apoptosis. To determine if DNA-PK and p53 interact to govern radiosensitivity, we compared the response of single and compound mutant mice to 8 Gy IR. Compound mutant Prkdc(scid/scid)/Trp53(-/-)mice died earliest due to severe GI-ARS. While both Prkdc(scid/scid) and Prkdc(scid/scid)/Trp53(-/-)mutant mice had higher levels of IR-induced DNA damage, particularly within the stem cell compartment of the intestinal crypt, in Prkdc(scid/scid)/Trp53(-/-)mice these damaged cells abnormally progressed through the cell cycle resulting in mitotic cell death. This led to a loss of Paneth cells and a failure to regenerate the differentiated epithelial cells required for intestinal function. IR-induced apoptosis did not correlate with radiosensitivity. Overall, these data reveal that DNA repair, mediated by DNA-PK, and cell cycle arrest, mediated by p53, cooperate to protect the stem cell niche after DNA damage, suggesting combination approaches to modulate both pathways may be beneficial to reduce GI-ARS. As many cancers harbor p53 mutations, this also suggests targeting DNA-PK may be effective to enhance sensitivity of p53 mutant tumors to radiation.

  12. A direct comparison of protein structure in the gas and solution phase: the Trp-cage

    DEFF Research Database (Denmark)

    Patriksson, Alexandra; Adams, Christopher M; Kjeldsen, Frank;

    2007-01-01

    Molecular dynamics simulations of zwitterions of the Trp-cage protein in the gas phase show that the most stable ion in vacuo has preserved the charge locations acquired in solution. A direct comparison of the gas and solution-phase structures reveals that, despite the similarity in charge location......, there is significant difference in the structures, with a substantial increase in hydrogen bonds and exposure of hydrophobic parts in the gas phase. The structure of the salt bridge in the gas phase is also much more stable than in the (experimental) solution structure....

  13. Lipid Storage Disorders Block Lysosomal Trafficking By Inhibiting TRP Channel and Calcium Release

    OpenAIRE

    2012-01-01

    Lysosomal lipid accumulation, defects in membrane trafficking, and altered Ca2+ homeostasis are common features in many lysosomal storage diseases. Mucolipin TRP channel 1 (TRPML1) is the principle Ca2+ channel in the lysosome. Here we show that TRPML1-mediated lysosomal Ca2+ release, measured using a genetically-encoded Ca2+ indicator (GCaMP3) attached directly to TRPML1 and elicited by a potent membrane-permeable synthetic agonist, is dramatically reduced in Niemann-Pick (NP) disease cells....

  14. Thioredoxin Txnl1/TRP32 Is a Redox-active Cofactor of the 26 S Proteasome

    DEFF Research Database (Denmark)

    Andersen, Katrine M; Klausen, Louise Kjær; Prag, Søren

    2009-01-01

    The 26S proteasome is a large proteolytic machine, which degrades most intracellular proteins. We found that thioredoxin, Txnl1/TRP32, binds to Rpn11, a subunit of the regulatory complex of the human 26S proteasome. Txnl1 is abundant, metabolically stable and widely expressed and is present...... in the cytoplasm and nucleus. Txnl1 has thioredoxin activity with a redox potential of about -250 mV. Mutant Txnl1 with one active site cysteine replaced by serine formed disulfide bonds to eEF1A1, a substrate-recruiting factor of the 26S proteasome. eEF1A1 is therefore a likely physiological substrate...

  15. Mecanoproteínas TRP(transient-receptor potencial) y movimiento dentario

    OpenAIRE

    Broseta Collado, Laura

    2012-01-01

    Algunos miembros de la familia de los canales iónicos TRP (transient receptor potential) funcionan como proteínas en una amplia variedad de tejidos y células. En el presente estudio se han utilizado técnicas de inmunohistoquímica para analizar la presencia de la mecano-proteína TRPV4 en el ligamento periodontal del primer molar superior de la rata en condiciones de normalidad y sometido a tracción ortodóncica (50 g) durante 10 días. En el diente control la inmunorreacción para TRPV4 es escasa...

  16. The open-close mechanism of M2 channel protein in influenza A virus: A computational study on the hydrogen bonds and cation-π interactions among His37 and Trp41

    Institute of Scientific and Technical Information of China (English)

    CHENG JiaGao; ZHU WeiLiang; WANG YanLi; YAN XiuHua; LI Zhong; TANG Yun; JIANG HuaLiang

    2008-01-01

    The M2 protein from influenza A virus is a tetrameric ion channel. It was reported that the permeation of the ion channel is correlated with the hydrogen bond network among His37 residues and the cation-π interactions between His37 and Trp41. In the present study, the hydrogen bonding network of 4-methyl-imidazoles was built to mimic the hydrogen bonds between His37 residues, and the cation-π interactions between 4-methyl-imidazolium and indole systems were selected to represent the interac-tions between His37 and Trp41. Then, quantum chemistry calculations at the MP2/6-311G** level were carried out to explore the properties of the hydrogen bonds and the cation-π interactions. The calculation results indicate that the binding strength of the N--H…N hydrogen bond between imidazole rings is up to -6.22 kcal·mol-1, and the binding strength of the strongest cation-π interaction is up to -18.8 kcal·mol-1 (T-shaped interaction) or -12.3 kcal·mol-1 (parallel stacking interaction). Thus, the calculated binding energies indicate that it is possible to control the permeation of the M2 ion channel through the hydrogen bond network and the cation-π interactions by altering the pH values.

  17. The open-close mechanism of M2 channel protein in influenza A virus:A computational study on the hydrogen bonds and cation-π interactions among His37 and Trp41

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The M2 protein from influenza A virus is a tetrameric ion channel. It was reported that the permeation of the ion channel is correlated with the hydrogen bond network among His37 residues and the cation-π interactions between His37 and Trp41. In the present study,the hydrogen bonding network of 4-methyl-imidazoles was built to mimic the hydrogen bonds between His37 residues,and the cation-π interactions between 4-methyl-imidazolium and indole systems were selected to represent the interac-tions between His37 and Trp41. Then,quantum chemistry calculations at the MP2/6-311G level were carried out to explore the properties of the hydrogen bonds and the cation-π interactions. The calcula-tion results indicate that the binding strength of the N-H···N hydrogen bond between imidazole rings is up to -6.22 kcal·mol-1,and the binding strength of the strongest cation-π interaction is up to -18.8 kcal·mol-1(T-shaped interaction) or -12.3 kcal·mol-1(parallel stacking interaction). Thus,the calcu-lated binding energies indicate that it is possible to control the permeation of the M2 ion channel through the hydrogen bond network and the cation-π interactions by altering the pH values.

  18. Towards understanding the tandem mass spectra of protonated oligopeptides. 2: The proline effect in collision-induced dissociation of protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp).

    Science.gov (United States)

    Bleiholder, Christian; Suhai, Sándor; Harrison, Alex G; Paizs, Béla

    2011-06-01

    The product ion spectra of proline-containing peptides are commonly dominated by y(n) ions generated by cleavage at the N-terminal side of proline residues. This proline effect is investigated in the current work by collision-induced dissociation (CID) of protonated Ala-Ala-Xxx-Pro-Ala (Xxx includes Ala, Ser, Leu, Val, Phe, and Trp) in an electrospray/quadrupole/time-of-flight (QqTOF) mass spectrometer and by quantum chemical calculations on protonated Ala-Ala-Ala-Pro-Ala. The CID spectra of all investigated peptides show a dominant y(2) ion (Pro-Ala sequence). Our computational results show that the proline effect mainly arises from the particularly low threshold energy for the amide bond cleavage N-terminal to the proline residue, and from the high proton affinity of the proline-containing C-terminal fragment produced by this cleavage. These theoretical results are qualitatively supported by the experimentally observed y(2)/b(3) abundance ratios for protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp). In the post-cleavage phase of fragmentation the N-terminal oxazolone fragment with the Ala-Ala-Xxx sequence and Pro-Ala compete for the ionizing proton for these peptides. As the proton affinity of the oxazolone fragment increases, the y(2)/b(3) abundance ratio decreases.

  19. Towards Understanding the Tandem Mass Spectra of Protonated Oligopeptides. 2: The Proline Effect in Collision-Induced Dissociation of Protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp)

    Science.gov (United States)

    Bleiholder, Christian; Suhai, Sándor; Harrison, Alex G.; Paizs, Béla

    2011-06-01

    The product ion spectra of proline-containing peptides are commonly dominated by y n ions generated by cleavage at the N-terminal side of proline residues. This proline effect is investigated in the current work by collision-induced dissociation (CID) of protonated Ala-Ala-Xxx-Pro-Ala (Xxx includes Ala, Ser, Leu, Val, Phe, and Trp) in an electrospray/quadrupole/time-of-flight (QqTOF) mass spectrometer and by quantum chemical calculations on protonated Ala-Ala-Ala-Pro-Ala. The CID spectra of all investigated peptides show a dominant y 2 ion (Pro-Ala sequence). Our computational results show that the proline effect mainly arises from the particularly low threshold energy for the amide bond cleavage N-terminal to the proline residue, and from the high proton affinity of the proline-containing C-terminal fragment produced by this cleavage. These theoretical results are qualitatively supported by the experimentally observed y 2 / b 3 abundance ratios for protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp). In the post-cleavage phase of fragmentation the N-terminal oxazolone fragment with the Ala-Ala-Xxx sequence and Pro-Ala compete for the ionizing proton for these peptides. As the proton affinity of the oxazolone fragment increases, the y 2 / b 3 abundance ratio decreases.

  20. Influence of glycine residues on peptide conformation in membrane environments.

    Science.gov (United States)

    Li, S C; Deber, C M

    1992-01-01

    Transmembrane (TM) segments of integral membrane proteins are putatively alpha-helical in conformation, yet their primary sequences are rich in residues known in globular proteins as helix-breakers (Gly) and beta-sheet promoters (Ile, Val, Thr). To examine the specific 2 degrees structure propensities of such residues in membrane environments, we have now designed and synthesized a series of model 20-residue peptides with "guest" hydrophobia segments embedded in "host" N- and C-terminal hydrophilic matrices. Molecular design was based on the prototypical sequence NH2-(Ser-Lys)2-Ala5-Leu6-x7-Ala8-Leu9-y10-Trp 11-Ala12-Leu13-z14-(Lys-Ser)3-OH. The 10-residue hydrophobic mid-segment 5-14 is expected to act as ca. three turns of an alpha-helix. In the present work, we compare the 20-residue peptide having three "helix-forming" Ala residues [x = y = z = Ala (peptide 3A)] to the corresponding peptide 3G (x = y = z = Gly) which contains three "helix-breaking" Gly residues. Trp was inserted to provide a measure of aromatic character typical of TM segments; Ser and Lys enhanced solubility in aqueous media. Circular dichroism studies in water, in a membrane-mimetic [sodium dodecylsulfate (SDS)], medium, and in methanol solutions, demonstrated the exquisite sensitivity of the conformations of these peptides to environment, and proved that despite its backbone flexibility, Gly can be accommodated as readily as Ala into a hydrophobic alpha-helix in a membrane. Nevertheless, the relative stability of Ala- vs. Gly-containing helices emerged in methanol solvent titration and temperature dependence experiments in SDS.(ABSTRACT TRUNCATED AT 250 WORDS)

  1. Structural Analysis of the Streptomyces avermitilis CYP107W1-Oligomycin A Complex and Role of the Tryptophan 178 Residue.

    Science.gov (United States)

    Han, Songhee; Pham, Tan-Viet; Kim, Joo-Hwan; Lim, Young-Ran; Park, Hyoung-Goo; Cha, Gun-Su; Yun, Chul-Ho; Chun, Young-Jin; Kang, Lin-Woo; Kim, Donghak

    2016-03-01

    CYP107W1 from Streptomyces avermitilis is a cytochrome P450 enzyme involved in the biosynthesis of macrolide oligomycin A. A previous study reported that CYP107W1 regioselectively hydroxylated C12 of oligomycin C to produce oligomycin A, and the crystal structure of ligand free CYP107W1 was determined. Here, we analyzed the structural properties of the CYP107W1-oligomycin A complex and characterized the functional role of the Trp178 residue in CYP107W1. The crystal structure of the CYP107W1 complex with oligomycin A was determined at a resolution of 2.6 Å. Oligomycin A is bound in the substrate access channel on the upper side of the prosthetic heme mainly by hydrophobic interactions. In particular, the Trp178 residue in the active site intercalates into the large macrolide ring, thereby guiding the substrate into the correct binding orientation for a productive P450 reaction. A Trp178 to Gly mutation resulted in the distortion of binding titration spectra with oligomycin A, whereas binding spectra with azoles were not affected. The Gly178 mutant's catalytic turnover number for the 12-hydroxylation reaction of oligomycin C was highly reduced. These results indicate that Trp178, located in the open pocket of the active site, may be a critical residue for the productive binding conformation of large macrolide substrates.

  2. Two key residues in ephrinB3 are critical for its use as an alternative receptor for Nipah virus.

    Directory of Open Access Journals (Sweden)

    Oscar A Negrete

    2006-02-01

    Full Text Available EphrinB2 was recently discovered as a functional receptor for Nipah virus (NiV, a lethal emerging paramyxovirus. Ephrins constitute a class of homologous ligands for the Eph class of receptor tyrosine kinases and exhibit overlapping expression patterns. Thus, we examined whether other ephrins might serve as alternative receptors for NiV. Here, we show that of all known ephrins (ephrinA1-A5 and ephrinB1-B3, only the soluble Fc-fusion proteins of ephrinB3, in addition to ephrinB2, bound to soluble NiV attachment protein G (NiV-G. Soluble NiV-G bound to cell surface ephrinB3 and B2 with subnanomolar affinities (Kd = 0.58 nM and 0.06 nM for ephrinB3 and B2, respectively. Surface plasmon resonance analysis indicated that the relatively lower affinity of NiV-G for ephrinB3 was largely due to a faster off-rate (K(off = 1.94 x 10(-3 s(-1 versus 1.06 x 10(-4 s(-1 for ephrinB3 and B2, respectively. EphrinB3 was sufficient to allow for viral entry of both pseudotype and live NiV. Soluble ephrinB2 and B3 were able to compete for NiV-envelope-mediated viral entry on both ephrinB2- and B3-expressing cells, suggesting that NiV-G interacts with both ephrinB2 and B3 via an overlapping site. Mutational analysis indicated that the Leu-Trp residues in the solvent exposed G-H loop of ephrinB2 and B3 were critical determinants of NiV binding and entry. Indeed, replacement of the Tyr-Met residues in the homologous positions in ephrinB1 with Leu-Trp conferred NiV receptor activity to ephrinB1. Thus, ephrinB3 is a bona fide alternate receptor for NiV entry, and two residues in the G-H loop of the ephrin B-class ligands are critical determinants of NiV receptor activity.

  3. Construction of pEGFP-ChEgTrp as DNA model for multi-epitope vaccine against Echinococcus granulosus

    Directory of Open Access Journals (Sweden)

    M. Ahmadzadeh

    2015-12-01

    Full Text Available Background: Infection with Echinococcus granulosus causes hydatidosis in human and ruminants. With regards to the high prevalence of hydatidosis in Iran, dealing with this disease is important in terms of public health. Objective: The aim of this study was to construct pEGFP-ChEgTrp as DNA model for multi-epitope vaccine against Echinococcus granulosus Methods: This experimental study was conducted in the Razi Vaccine & Serum Research Institute, Karaj in 2013. Initially, epitopes stimulating the host immune response were predicted by IEDB Database and the coding sequences were made. The sequences were amplified by PCR. The PCR products were cloned into pEGFP-N1 vector after digestion with XhoI restriction enzyme. The bacteria containing recombinant plasmid were evaluated using Colony PCR, agarose gel electrophoresis and sequencing methods. Findings: Four peptides with 10 linear epitopes were predicted in EgTrp antigen. The nucleotide sequence coding ChEgTrp was amplified by PCR using specific primers and a 270 bp fragment was obtained. This fragment was cloned into pEGFP-N1 vector and the recombinant plasmid was confirmed by Colony PCR and agarose gel electrophoresis. For final confirmation, the recombinant plasmid was sequenced and the pEGFP-ChEgTrp was constructed. Conclusion: The ChEgTrp was successfully cloned into the pEGFP-N1 vector and this plasmid can be used to design DNA vaccines.

  4. Brca2 and Trp53 deficiency cooperate in the progression of mouse prostate tumourigenesis.

    Directory of Open Access Journals (Sweden)

    Jeffrey C Francis

    2010-06-01

    Full Text Available Epidemiological studies have shown that one of the strongest risk factors for prostate cancer is a family history of the disease, suggesting that inherited factors play a major role in prostate cancer susceptibility. Germline mutations in BRCA2 predispose to breast and ovarian cancer with its predominant tumour suppressor function thought to be the repair of DNA double-strand breaks. BRCA2 has also been implicated in prostate cancer etiology, but it is unclear the impact that mutations in this gene have on prostate tumourigenesis. Here we have undertaken a genetic analysis in the mouse to determine the role of Brca2 in the adult prostate. We show that deletion of Brca2 specifically in prostate epithelia results in focal hyperplasia and low-grade prostate intraepithelial neoplasia (PIN in animals over 12 months of age. Simultaneous deletion of Brca2 and the tumour suppressor Trp53 in prostate epithelia gave rise to focal hyperplasia and atypical cells at 6 months, leading to high-grade PIN in animals from 12 months. Epithelial cells in these lesions show an increase in DNA damage and have higher levels of proliferation, but also elevated apoptosis. Castration of Brca2;Trp53 mutant animals led to regression of PIN lesions, but atypical cells persisted that continued to proliferate and express nuclear androgen receptor. This study provides evidence that Brca2 can act as a tumour suppressor in the prostate, and the model we describe should prove useful in the development of new therapeutic approaches.

  5. TRP channels and traffic-related environmental pollution-induced pulmonary disease.

    Science.gov (United States)

    Akopian, Armen N; Fanick, E Robert; Brooks, Edward G

    2016-05-01

    Environmental pollutant exposures are major risk factors for adverse health outcomes, with increased morbidity and mortality in humans. Diesel exhaust (DE) is one of the major harmful components of traffic-related air pollution. Exposure to DE affects several physiological systems, including the airways, and pulmonary diseases are increased in highly populated urban areas. Hence, there are urgent needs to (1) create newer and lesser polluting fuels, (2) improve exhaust aftertreatments and reduce emissions, and (3) understand mechanisms of actions for toxic effects of both conventional and cleaner diesel fuels on the lungs. These steps could aid the development of diagnostics and interventions to prevent the negative impact of traffic-related air pollution on the pulmonary system. Exhaust from conventional, and to a lesser extent, clean fuels, contains particulate matter (PM) and more than 400 additional chemical constituents. The major toxic constituents are nitrogen oxides (NOx) and polycyclic aromatic hydrocarbons (PAHs). PM and PAHs could potentially act via transient receptor potential (TRP) channels. In this review, we will first discuss the associations between DE from conventional as well as clean fuel technologies and acute and chronic airway inflammation. We will then review possible activation and/or potentiation of TRP vanilloid type 1 (TRPV1) and ankyrin 1 (TRPA1) channels by PM and PAHs. Finally, we will discuss and summarize recent findings on the mechanisms whereby TRPs could control the link between DE and airway inflammation, which is a primary determinant leading to pulmonary disease.

  6. Fast TRP/TIS Measurement Method for Notebook PC HSDPA%笔记本电脑HSDPA的快速TRP/TIS测量方法

    Institute of Scientific and Technical Information of China (English)

    李丹; 冯正和

    2008-01-01

    为了实现笔记本电脑中HSDPA天线及噪声的快速调试,对现有的测量方法进行了改进:在有源和无源方向图相似的条件下,利用三维无源测量的结果和单点有源测量的结果获得TRP和TIS的估算.通过和标准有源暗室的测量结果对比发现,该方法具有和三维有源测量方法可比拟的准确度,并且在这方面远好于单点峰值有源测量方法;另一方面,由于该测量方法仅仅包含三维无源测量、单点有源测量以及简单的计算,因此相对于三维有源测量而言,可以大大提高TRP和TIS的测量速度.基于上面的原因,本文提出的快速测量方法不失为当前TRP/TIS测量方法的有效补充.

  7. Preliminary Crystallography Confirms that the Archaeal DNA-binding and Tryptophan-sensing Regulator TrpY is a Dimer

    Energy Technology Data Exchange (ETDEWEB)

    J Cafasso; B Manjasetty; E Karr; K Sandman; M Chance; J Reeve

    2011-12-31

    TrpY regulates the transcription of the metabolically expensive tryptophan-biosynthetic operon in the thermophilic archaeon Methanothermobacter thermautotrophicus. TrpY was crystallized using the hanging-drop method with ammonium sulfate as the precipitant. The crystals belonged to the tetragonal space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 87, c = 147 {angstrom}, and diffracted to 2.9 {angstrom} resolution. The possible packing of molecules within the cell based on the values of the Matthews coefficient (V{sub M}) and analysis of the self-rotation function are consistent with the asymmetric unit being a dimer. Determining the structure of TrpY in detail will provide insight into the mechanisms of DNA binding, tryptophan sensing and transcription regulation at high temperature by this novel archaeal protein.

  8. Structural model of a putrescine-cadaverine permease from Trypanosoma cruzi predicts residues vital for transport and ligand binding.

    Science.gov (United States)

    Soysa, Radika; Venselaar, Hanka; Poston, Jacqueline; Ullman, Buddy; Hasne, Marie-Pierre

    2013-06-15

    The TcPOT1.1 gene from Trypanosoma cruzi encodes a high affinity putrescine-cadaverine transporter belonging to the APC (amino acid/polyamine/organocation) transporter superfamily. No experimental three-dimensional structure exists for any eukaryotic member of the APC family, and thus the structural determinants critical for function of these permeases are unknown. To elucidate the key residues involved in putrescine translocation and recognition by this APC family member, a homology model of TcPOT1.1 was constructed on the basis of the atomic co-ordinates of the Escherichia coli AdiC arginine/agmatine antiporter crystal structure. The TcPOT1.1 homology model consisted of 12 transmembrane helices with the first ten helices organized in two V-shaped antiparallel domains with discontinuities in the helical structures of transmembrane spans 1 and 6. The model suggests that Trp241 and a Glu247-Arg403 salt bridge participate in a gating system and that Asn245, Tyr148 and Tyr400 contribute to the putrescine-binding pocket. To test the validity of the model, 26 site-directed mutants were created and tested for their ability to transport putrescine and to localize to the parasite cell surface. These results support the robustness of the TcPOT1.1 homology model and reveal the importance of specific aromatic residues in the TcPOT1.1 putrescine-binding pocket.

  9. An association between TRP64ARG polymorphism of the B3 adrenoreceptor gene and some metabolic disturbances

    Directory of Open Access Journals (Sweden)

    Abilova Samai S

    2011-10-01

    Full Text Available Abstract Backgrounds B3 adrenoreceptors (ADRB3 are abundant in adipose tissue and play the role in its metabolism and lipolysis. Some variants of the ADRB3 gene may predispose subjects for the development obesity and metabolic abnormalities in the setting of modern sedentary lifestyle. ADRB3 gene polymorphism association with metabolic disturbances has never been studied before in the ethnic Kyrgyz population. Aim To study an association between Trp64Arg polymorphism of the ADRB3 and metabolic syndrome (MS components in an ethnic Kyrgyz group. Materials and methods 213 Ethnic Kyrgyz volunteers over the age of 30 were enrolled in the study. The assessment plan for each individual comprised of general physical and anthropometric exams as well as laboratory tests (glucose, lipid panel, insulin and genotyping by Trp64Arg polymorphism of the ADRB3. MS diagnosis was consistent with modified ATP III criteria (2005. Logistic regression analysis was performed to test the potential independent association between Arg64 allele with obesity, abdominal obesity (AO and arterial hypertension (AH. Results Trp64Arg polymorphism of the ADRB3 was assessed in 213 individuals (145 men, 68 women aged 30-73 (mean age 50.7 ± 7.6. Arg64 allele frequency was 0.239; ADRB3 genotype distribution among participants was: Trp64 homozygotes 54.5%, Trp64Arg 43.2% and Arg64 homozygotes 2.3%. There was an association between Trp64Arg и Arg64Arg genotypes and higher BMI, WC and obesity frequency (p Conclusion Arg64 allele of the ADRB3 gene in the studied group has an association with MS components such as obesity, AO and decreased HDL-C level.

  10. Ehrlichia chaffeensis TRP120 Activates Canonical Notch Signaling To Downregulate TLR2/4 Expression and Promote Intracellular Survival

    Directory of Open Access Journals (Sweden)

    Taslima T. Lina

    2016-07-01

    Full Text Available Ehrlichia chaffeensis preferentially targets mononuclear phagocytes and survives through a strategy of subverting innate immune defenses, but the mechanisms are unknown. We have shown E. chaffeensis type 1 secreted tandem repeat protein (TRP effectors are involved in diverse molecular pathogen-host interactions, such as the TRP120 interaction with the Notch receptor-cleaving metalloprotease ADAM17. In the present study, we demonstrate E. chaffeensis, via the TRP120 effector, activates the canonical Notch signaling pathway to promote intracellular survival. We found that nuclear translocation of the transcriptionally active Notch intracellular domain (NICD occurs in response to E. chaffeensis or recombinant TRP120, resulting in upregulation of Notch signaling pathway components and target genes notch1, adam17, hes, and hey. Significant differences in canonical Notch signaling gene expression levels (>40% were observed during early and late stages of infection, indicating activation of the Notch pathway. We linked Notch pathway activation specifically to the TRP120 effector, which directly interacts with the Notch metalloprotease ADAM17. Using pharmacological inhibitors and small interfering RNAs (siRNAs against γ-secretase enzyme, Notch transcription factor complex, Notch1, and ADAM17, we demonstrated that Notch signaling is required for ehrlichial survival. We studied the downstream effects and found that E. chaffeensis TRP120-mediated activation of the Notch pathway causes inhibition of the extracellular signal-regulated kinase 1/2 (ERK1/2 and p38 mitogen-activated protein kinase (MAPK pathways required for PU.1 and subsequent Toll-like receptor 2/4 (TLR2/4 expression. This investigation reveals a novel mechanism whereby E. chaffeensis exploits the Notch pathway to evade the host innate immune response for intracellular survival.

  11. XRCC1 Arg194Trp and Arg399Gln polymorphisms and arsenic methylation capacity are associated with urothelial carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Chiang, Chien-I [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Huang, Ya-Li [Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan (China); Chen, Wei-Jen [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Shiue, Horng-Sheng [Department of Chinese Medicine, Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taoyuan, Taiwan (China); Huang, Chao-Yuan; Pu, Yeong-Shiau [Department of Urology, National Taiwan University Hospital, College of Medicine National Taiwan University, Taipei, Taiwan (China); Lin, Ying-Chin [Department of Family Medicine, Shung Ho Hospital, Taipei Medical University, New Taipei, Taiwan (China); Department of Health Examination, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan (China); Division of Family Medicine, School of Medicine, Taipei Medical University, Taipei, Taiwan (China); Hsueh, Yu-Mei, E-mail: ymhsueh@tmu.edu.tw [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan (China)

    2014-09-15

    The association between DNA repair gene polymorphisms and bladder cancer has been widely studied. However, few studies have examined the correlation between urothelial carcinoma (UC) and arsenic or its metabolites. The aim of this study was to examine the association between polymorphisms of the DNA repair genes, XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln, with urinary arsenic profiles and UC. To this end, we conducted a hospital-based case–control study with 324 UC patients and 647 age- and gender-matched non-cancer controls. Genomic DNA was used to examine the genotype of XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln by PCR-restriction fragment length polymorphism analysis (PCR-RFLP). Urinary arsenic profiles were measured by high performance liquid chromatography (HPLC) linked with hydride generator and atomic absorption spectrometry. The XRCC1 399 Gln/Gln and 194 Arg/Trp and Trp/Trp genotypes were significantly related to UC, and the odds ratio (OR) and 95% confidence interval (95%CI) were 1.68 (1.03–2.75) and 0.66 (0.48–0.90), respectively. Participants with higher total urinary arsenic levels, a higher percentage of inorganic arsenic (InAs%) and a lower percentage of dimethylarsinic acid (DMA%) had a higher OR of UC. Participants carrying XRCC1 risk diplotypes G-C/G-C, A-C/A-C, and A-T/G-T, and who had higher total arsenic levels, higher InAs%, or lower DMA% compared to those with other XRCC1 diplotypes had a higher OR of UC. Our results suggest that the XRCC1 399 Gln/Gln and 194 Arg/Arg DNA repair genes play an important role in poor arsenic methylation capacity, thereby increasing the risk of UC in non-obvious arsenic exposure areas. - Highlights: • The XRCC1 399Gln/Gln genotype was significantly associated with increased OR of UC. • The XRCC1 194 Arg/Trp and Trp/Trp genotype had a significantly decreased OR of UC. • Combined effect of the XRCC1 genotypes and poor arsenic methylation capacity on

  12. The Transient Receptor Potential (TRP) Channel Family in Colletotrichum graminicola: A Molecular and Physiological Analysis.

    Science.gov (United States)

    Lange, Mario; Weihmann, Fabian; Schliebner, Ivo; Horbach, Ralf; Deising, Holger B; Wirsel, Stefan G R; Peiter, Edgar

    2016-01-01

    Calcium (Ca2+) is a universal second messenger in all higher organisms and centrally involved in the launch of responses to environmental stimuli. Ca2+ signals in the cytosol are initiated by the activation of Ca2+ channels in the plasma membrane and/or in endomembranes. Yeast (Saccharomyces cerevisiae) contains a Ca2+-permeable channel of the TRP family, TRPY1, which is localized in the vacuolar membrane and contributes to cytosolic free Ca2+ ([Ca2+]cyt) elevations, for example in response to osmotic upshock. A TRPY1 homologue in the rice blast fungus is known to be important for growth and pathogenicity. To determine the role of the TRP channel family in the maize pathogen Colletotrichum graminicola, proteins homologous to TRPY1 were searched. This identified not one, but four genes in the C. graminicola genome, which had putative orthologs in other fungi, and which we named CgTRPF1 through 4. The topology of the CgTRPF proteins resembled that of TRPY1, albeit with a variable number of transmembrane (TM) domains additional to the six-TM-domain core and a diverse arrangement of putatively Ca2+-binding acidic motifs. All CgTRPF genes were expressed in axenic culture and throughout the infection of maize. Like TRPY1, all TRPF proteins of C. graminicola were localized intracellularly, albeit three of them were found not in large vacuoles, but co-localized in vesicular structures. Deletion strains for the CgTRPF genes were not altered in processes thought to involve Ca2+ release from internal stores, i.e. spore germination, the utilization of complex carbon sources, and the generation of tip-focussed [Ca2+]cyt spikes. Heterologous expression of CgTRPF1 through 4 in a tryp1Δ yeast mutant revealed that none of the channels mediated the release of Ca2+ in response to osmotic upshock. Accordingly, aequorin-based [Ca2+]cyt measurements of C. graminicola showed that in this fungus, osmotic upshock-triggered [Ca2+]cyt elevations were generated entirely by influx of Ca2

  13. The Transient Receptor Potential (TRP Channel Family in Colletotrichum graminicola: A Molecular and Physiological Analysis.

    Directory of Open Access Journals (Sweden)

    Mario Lange

    Full Text Available Calcium (Ca2+ is a universal second messenger in all higher organisms and centrally involved in the launch of responses to environmental stimuli. Ca2+ signals in the cytosol are initiated by the activation of Ca2+ channels in the plasma membrane and/or in endomembranes. Yeast (Saccharomyces cerevisiae contains a Ca2+-permeable channel of the TRP family, TRPY1, which is localized in the vacuolar membrane and contributes to cytosolic free Ca2+ ([Ca2+]cyt elevations, for example in response to osmotic upshock. A TRPY1 homologue in the rice blast fungus is known to be important for growth and pathogenicity. To determine the role of the TRP channel family in the maize pathogen Colletotrichum graminicola, proteins homologous to TRPY1 were searched. This identified not one, but four genes in the C. graminicola genome, which had putative orthologs in other fungi, and which we named CgTRPF1 through 4. The topology of the CgTRPF proteins resembled that of TRPY1, albeit with a variable number of transmembrane (TM domains additional to the six-TM-domain core and a diverse arrangement of putatively Ca2+-binding acidic motifs. All CgTRPF genes were expressed in axenic culture and throughout the infection of maize. Like TRPY1, all TRPF proteins of C. graminicola were localized intracellularly, albeit three of them were found not in large vacuoles, but co-localized in vesicular structures. Deletion strains for the CgTRPF genes were not altered in processes thought to involve Ca2+ release from internal stores, i.e. spore germination, the utilization of complex carbon sources, and the generation of tip-focussed [Ca2+]cyt spikes. Heterologous expression of CgTRPF1 through 4 in a tryp1Δ yeast mutant revealed that none of the channels mediated the release of Ca2+ in response to osmotic upshock. Accordingly, aequorin-based [Ca2+]cyt measurements of C. graminicola showed that in this fungus, osmotic upshock-triggered [Ca2+]cyt elevations were generated entirely by

  14. 6-Nitro-L-tryptophan: a novel spectroscopic probe of trp aporepressor and human serum albumin.

    Science.gov (United States)

    Phillips, R S; Marmorstein, R Q

    1988-04-01

    The binding of 6-nitro-L-tryptophan to trp aporepressor and human serum albumin has been examined by visible difference spectroscopy and circular dichroism. 6-Nitro-L-tryptophan, prepared by nitration of L-tryptophan with nitric acid in glacial acetic acid, exhibits a visible and near-uv absorption spectrum with lambda max at about 330 nm (epsilon = 7 X 10(3) M-1 cm-1) and a shoulder near 380 nm in H2O. In the presence of trp aporepressor, the visible absorption intensity is sharply diminished. Visible difference spectral titration data give KD = 1.27 X 10(-4) M and n = 0.95 per subunit at 25 degrees C. While 6-nitro-L-tryptophan exhibits no significant circular dichroism between 300 and 500 nm, the complex with trp aporepressor exhibits strong circular dichroism signals, with a negative maximum at 386 nm (delta epsilon = -7.5 M-1 cm-1) and a positive maximum at 310 nm (delta epsilon = +6 M-1 cm-1). Circular dichroism titration data give KD = 1.69 X 10(-4) M and n = 0.90 per subunit at 25 degrees C. The KD values determined spectroscopically are in excellent agreement with that determined by equilibrium dialysis, KD = 1.5 X 10(-4) M at 25 degrees C. In the presence of human serum albumin, the spectrum of 6-nitro-L-tryptophan exhibits a blue shift and an increase in absorption intensity; similar changes are observed in solvents of low dielectric contrast such as 80% aqueous dioxane. Visible difference spectral titration data give KD = 8.0 X 10(-5) M and n = 0.95 for human serum albumin. The complex of 6-nitro-L-tryptophan with human serum albumin exhibits a strong positive circular dichroism maximum at 380 nm (delta epsilon = +9.8 M-1 cm-1) with a shoulder at 310-320 nm. Circular dichroism titration data give KD = 6.4 X 10(-5) M and n = 0.83, in good agreement with the visible difference spectral results. Taken together, our results demonstrate the utility of 6-nitro-L-tryptophan as a spectroscopic probe for tryptophan-binding proteins.

  15. Alanine scanning mutagenesis of anti-TRAP (AT) reveals residues involved in binding to TRAP.

    Science.gov (United States)

    Chen, Yanling; Gollnick, Paul

    2008-04-11

    The trp RNA-binding attenuation protein (TRAP) regulates expression of the tryptophan biosynthetic (trp) genes in response to changes in intracellular levels of free l-tryptophan in many Gram-positive bacteria. When activated by binding tryptophan, TRAP binds to the mRNAs of several genes involved in tryptophan metabolism, and down-regulates transcription or translation of these genes. Anti-TRAP (AT) is an antagonist of TRAP that binds to tryptophan-activated TRAP and prevents it from binding to its RNA targets, and thereby up-regulates trp gene expression. The crystal structure shows that AT is a cone-shaped trimer (AT(3)) with the N-terminal residues of the three subunits assembled at the apex of the cone and that these trimers can further assemble into a dodecameric (AT(12)) structure. Using alanine-scanning mutagenesis we found four residues, all located on the "top" region of AT(3), that are essential for binding to TRAP. Fluorescent labeling experiments further suggest that the top region of AT is in close juxtaposition to TRAP in the AT-TRAP complex. In vivo studies confirmed the importance of these residues on the top of AT in regulating TRAP mediated gene regulation.

  16. The π Configuration of the WWW Motif of a Short Trp-Rich Peptide Is Critical for Targeting Bacterial Membranes, Disrupting Preformed Biofilms, and Killing Methicillin-Resistant Staphylococcus aureus.

    Science.gov (United States)

    Zarena, D; Mishra, Biswajit; Lushnikova, Tamara; Wang, Fangyu; Wang, Guangshun

    2017-08-08

    Tryptophan-rich peptides, being short and suitable for large-scale chemical synthesis, are attractive candidates for developing a new generation of antimicrobials to combat antibiotic-resistant bacteria (superbugs). Although there are numerous pictures of the membrane-bound structure of a single tryptophan (W), how multiple Trp amino acids assemble themselves and interact with bacterial membranes is poorly understood. This communication presents the three-dimensional structure of an eight-residue Trp-rich peptide (WWWLRKIW-NH2 with 50% W) determined by the improved two-dimensional nuclear magnetic resonance method, which includes the measurements of (13)C and (15)N chemical shifts at natural abundance. This peptide forms the shortest two-turn helix with a distinct amphipathic feature. A unique structural arrangement is identified for the Trp triplet, WWW, that forms a π configuration with W2 as the horizontal bar and W1/W3 forming the two legs. An arginine scan reveals that the WWW motif is essential for killing methicillin-resistant Staphylococcus aureus USA300 and disrupting preformed bacterial biofilms. This unique π configuration for the WWW motif is stabilized by aromatic-aromatic interactions as evidenced by ring current shifts as well as nuclear Overhauser effects. Because the WWW motif is maintained, a change of I7 to R led to a potent antimicrobial and antibiofilm peptide with 4-fold improvement in cell selectivity. Collectively, this study elucidated the structural basis of antibiofilm activity of the peptide, identified a better peptide candidate via structure-activity relationship studies, and laid the foundation for engineering future antibiotics based on the WWW motif.

  17. Functional and topological studies with Trp-containing analogs of the peptide StII(1-30) derived from the N-terminus of the pore forming toxin sticholysin II: contribution to understand its orientation in membrane.

    Science.gov (United States)

    Ros, Uris; Souto, Ana Lucia C F; de Oliveira, Felipe J; Crusca, Edson; Pazos, Fabiola; Cilli, Eduardo M; Lanio, Maria E; Schreier, Shirley; Alvarez, Carlos

    2013-07-01

    Sticholysin II (St II) is the most potent cytolysin produced by the sea anemone Stichodactyla helianthus, exerting hemolytic activity via pore formation in membranes. The toxin's N-terminus contains an amphipathic α-helix that is very likely involved in pore formation. We have previously demonstrated that the synthetic peptide StII(1-30) encompassing the 1-30 segment of St II forms pores of similar radius to that of the protein (around 1 nm), being a good model of toxin functionality. Here we have studied the functional and conformational properties of fluorescent analogs of StII(1-30) in lipid membranes. The analogs were obtained by replacing Leu residues at positions 2, 12, 17, and 24 with the intrinsically fluorescent amino acid Trp (StII(1-30L2W), StII(1-30L12W), StII(1-30L17W), or StII(1-30L24W), respectively). The exchange by Trp did not significantly modify the activity and conformation of the parent peptide. The blue-shift and intensity enhancement of fluorescence in the presence of membrane indicated that Trp at position 2 is more deeply buried in the hydrophobic region of the bilayer. These experiments, as well as assays with water-soluble or spin-labeled lipid-soluble fluorescence quenchers suggest an orientation of StII(1-30) with its N-terminus oriented towards the hydrophobic core of the bilayer while the rest of the peptide is more exposed to the aqueous environment, as hypothesized for sticholysins.

  18. A neutralizing epitope of human papillomavirus type 11 is principally described by a continuous set of residues which overlap a distinct linear, surface-exposed epitope.

    Science.gov (United States)

    Ludmerer, S W; Benincasa, D; Mark, G E; Christensen, N D

    1997-01-01

    A panel of monoclonal antibodies (MAbs) which neutralize human papillomavirus type 11 (HPV11) in the athymic mouse xenograph neutralization assay and bind HPV11 virus-like particles (VLPs) has been described. We recently presented evidence that the Gly131-Tyr132 residues of the major capsid protein L1 confer type 11-specific binding. However, residues distally located on the primary L1 sequence also were shown to affect binding. This poses the question whether the epitope is principally centered in the region of Gly131-Tyr132 or, alternatively, is comprised of diversely located residues which come into proximity only upon proper assembly. We analyzed the result of numerous substitutions located between Tyr123 and Val142 of the HPV11 L1 sequence. We show that substitutions at five positions result in loss of binding for one or more of these MAbs by an enzyme-linked immunosorbent assay which measures antibody binding to VLPs. We demonstrate that binding of these MAbs is redirected to HPV16 VLPs which harbor eight type 11-like substitutions within the homologous region. Three of these substitutions did not affect binding when individually substituted in HPV11 but yet were still required to transfer binding to substituted HPV16 VLPs. The results demonstrate that the epitope for this class of neutralizing MAbs, although conformational and requiring VLP assembly for presentation, principally lies along a 20-residue stretch of the L1 major capsid protein. This targets the region for evaluation of the possibility of receptor binding and suggests possibilities for the design of peptide inhibitors of virus infectivity. PMID:9094659

  19. residue and shunting pinholes

    Science.gov (United States)

    Gorji, Nima E.

    2014-09-01

    The present work considers two observable phenomena through the experimental fabrication and electrical characterization of the rf-sputtered CdS/CdTe thin film solar cells that extremely reduce the overall conversion efficiency of the device: CdCl2 residue on the surface of the semiconductor and shunting pinholes. The former happens through nonuniform treatment of the As-deposited solar cells before annealing at high temperature and the latter occurs by shunting pinholes when the cell surface is shunted by defects, wire-like pathways or scratches on the metallic back contact caused from the external contacts. Such physical problems may be quite common in the experimental activities and reduce the performance down to 4-5 % which leads to dismantle the device despite its precise fabrication. We present our electrical characterization on the samples that received wet CdCl2 surface treatment (uniform or nonuniform) and are damaged by the pinholes.

  20. TRICK or TRP? What Trpc2-/- Mice Tell Us about Vomeronasal Organ Mediated Innate Behaviors

    Directory of Open Access Journals (Sweden)

    C. Ron eYu

    2015-06-01

    Full Text Available The vomeronasal organ (VNO plays an important role in mediating semiochemical communications and social behaviors in terrestrial species. Genetic knockout of individual components in the signaling pathways have been used to probe vomeronasal functions, and have provided much insights into how the VNO orchestrates innate behaviors. However, all data do not agree. In particular, knocking out Trpc2, a member of the TRP family of non-selective cationic channel thought to be the main transduction channel in the VNO, results in a number of fascinating behavioral phenotypes that have not been observed in other animals whose vomeronasal function is disrupted. Recent studies have identified signaling pathways that operate in parallel of Trpc2, raising the possibility that Trpc2 mutant animals may display neomorphic behaviors. In this article, I provide a critical analysis of emerging evidence to reconcile the discrepancies and discuss their implications.

  1. Angiotensin I-converting enzyme mutation (Trp1197Stop causes a dramatic increase in blood ACE.

    Directory of Open Access Journals (Sweden)

    Andrew B Nesterovitch

    Full Text Available BACKGROUND: Angiotensin-converting enzyme (ACE metabolizes many peptides and plays a key role in blood pressure regulation and vascular remodeling. Elevated ACE levels may be associated with an increased risk for different cardiovascular or respiratory diseases, including asthma. Previously, a molecular mechanism underlying a 5-fold familial increase of blood ACE was discovered: Pro1199Leu substitution enhanced the cleavage-secretion process. Carriers of this mutation were Caucasians from Europe (mostly Dutch or had European roots. METHODOLOGY/PRINCIPAL FINDINGS: We have found a family of African-American descent whose affected members' blood ACE level was increased 13-fold over normal. In affected family members, codon TGG coding for Trp1197 was substituted in one allele by TGA (stop codon. As a result, half of ACE expressed in these individuals had a length of 1196 amino acids and lacked a transmembrane anchor. This ACE mutant is not trafficked to the cell membrane and is directly secreted out of cells; this mechanism apparently accounts for the high serum ACE level seen in affected individuals. A haplotype of the mutant ACE allele was determined based on 12 polymorphisms, which may help to identify other carriers of this mutation. Some but not all carriers of this mutation demonstrated airflow obstruction, and some but not all have hypertension. CONCLUSIONS/SIGNIFICANCE: We have identified a novel Trp1197Stop mutation that results in dramatic elevation of serum ACE. Since blood ACE elevation is often taken as a marker of disease activity (sarcoidosis and Gaucher diseases, it is important for clinicians and medical scientists to be aware of alternative genetic causes of elevated blood ACE that are not apparently linked to disease.

  2. Tumor endothelial expression of P-glycoprotein upon microvesicular transfer of TrpC5 derived from adriamycin-resistant breast cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Dong, YePing; Pan, QiongXi; Jiang, Li; Chen, Zhen; Zhang, FangFang; Liu, YanJun; Xing, Hui; Shi, Mei; Li, Jiao; Li, XiYuan; Zhu, YaoDan; Chen, Yun; Bruce, Iain C.; Jin, Jian, E-mail: jinjian31@126.com; Ma, Xin, E-mail: maxin@jiangnan.edu.cn

    2014-03-28

    Highlights: • TrpC5 was mainly accumulated in microvesicles of drug-resistant MCF-7/ADM cells. • Microvesicles from MCF-7/ADM transferred TrpC5 to endothelial cells. • TrpC5 inhibition reduced P-glycoprotein accumulation on tumor blood vessels in vivo. - Abstract: Treatment of carcinoma commonly fails due to chemoresistance. Studies have shown that endothelial cells acquire resistance via the tumor microenvironment. Microvesicle (MV) shedding from the cell membrane to the microenvironment plays an important role in communication between cells. The aim of the present study was to determine whether MCF-7 adriamycin-resistant cells (MCF-7/ADM) shed MVs that alter the characteristics of human microvessel endothelial cells (HMECs). MVs from tumor cells transferred a Ca{sup 2+}-permeable channel TrpC5 to HMECs, inducing the expression of P-glycoprotein (P-gp) by activation of the transcription factor NFATc3 (nuclear factor of activated T cells isoform c3). Expression of the mdr1 gene was blocked by the TrpC5-blocking antibody T5E3, and the production of P-gp in HMECs was reduced by blockade of TrpC5. Thus, we postulate that endothelial cells acquire the resistant protein upon exposure to TrpC5-containg MVs in the microenvironment, and express P-gp in the TrpC5–NFATc3 signal pathway.

  3. Identification of a mutation in the tyrosinase related protein 1 (TRP1) gene associated with brown oculocutaneous albinism (OCA3)

    Energy Technology Data Exchange (ETDEWEB)

    Wildenberg, S.C.; Oetting, W.S.; Fryer, J.P. [Univ. of Minnesota, Minneapolis, MN (United States)] [and others

    1994-09-01

    The genes responsible for the two most common types of human oculocutaneous albinism (OCA) have been identified. Mutations of the tyrosinase gene (chromosome 11q14-21) produce OCA1, and mutations of the P gene (chromosome 15q11.2-13) produce OCA2. Another type of OCA known as brown OCA or OCA3 is found commonly in the African and African-American population. OCA3 is characterized by light brown skin and hair with the ocular features of albinism and represents the third most frequent type of OCA. We previously identified dizygotic African-American twin boys who were discordant for OCA3. Melanocytes from the affected twin produced brown melanin and contained no detectable TRP1 protein. We have now characterized the TRP1 gene from the affected twin. The human TRP1 gene, homologous to the murine brown locus, contains 8 exons and maps to chromosome 9p23. Using PCR amplification of each exon coupled with SSCP analysis and direct DNA sequencing, we found the affected twin to homozygous for a single bp deletion in exon 6. The deletion removes a G in codon 368 leading to a premature stop at codon 384. We also identified a Tsp509 polymorphism in the 3{prime} UTR. We conclude that mutations of the TRP1 gene are responsible for brown OCA or OCA3, making this the third major OCA gene identified in humans.

  4. Pubertal and adult windows of susceptibility to a high animal fat diet in Trp53-null mammary tumorigenesis.

    Science.gov (United States)

    Zhu, Yirong; Aupperlee, Mark D; Zhao, Yong; Tan, Ying Siow; Kirk, Erin L; Sun, Xuezheng; Troester, Melissa A; Schwartz, Richard C; Haslam, Sandra Z

    2016-12-13

    Premenopausal breast cancer is associated with increased animal fat consumption among normal weight, but not overweight women (Farvid et al., 2014). Our previous findings in obesity-resistant BALB/c mice similarly showed promotion of carcinogen-induced mammary tumorigenesis by a diet high in saturated animal fat (HFD). This effect was specific to pubertal versus adult HFD. This study identifies the effects of HFD during puberty versus adulthood in Trp53-null transplant BALB/c mice and investigates its mechanism of enhancing tumorigenesis. Either pubertal or adult HFD is sufficient to increase incidence of Trp53-null mammary tumors. Puberty-restricted HFD exposure promoted tumor cell proliferation, increased angiogenesis, and increased recruitment of total and M2 macrophages in epithelial tumors. Adult-restricted exposure to HFD similarly increased proliferation, angiogenesis, recruitment of total and M2 macrophages, and additionally reduced apoptosis. Adult HFD also increased incidence of spindle cell carcinomas resembling claudin-low breast cancer, and thus adult HFD in the Trp53-null transplantation system may be a useful model for human claudin low breast cancer. Importantly, these results on Trp53-null and our prior studies on DMBA-induced mammary tumorigenesis demonstrate a pubertal window of susceptibility to the promotional effects of HFD, indicating the potential of early life dietary intervention to reduce breast cancer risk.

  5. Solid-Phase Synthesis of New Trp(Nps-Containing Dipeptide Derivatives as TRPV1 Channel Blockers

    Directory of Open Access Journals (Sweden)

    Rosario González-Muñiz

    2010-07-01

    Full Text Available Trp(Nps-Lys-NH2 derivatives, bearing alkyl or guanidine groups either at the N–terminus or on the Lys side-chain or at both positions were conveniently prepared on solid-phase and evaluated as TRPV1 channel antagonists.

  6. Escoamento superficial e desagregação do solo em entressulcos em solo franco-argilo-arenoso com resíduos vegetais Interrill surface runoff and soil detachment on a sandy clay loam soil with residue cover

    Directory of Open Access Journals (Sweden)

    Elemar Antonino Cassol

    2004-07-01

    Full Text Available A presença de resíduos vegetais sobre a superfície do solo altera as características do escoamento superficial gerado pela chuva e a desagregação e transporte de sedimento resultantes do processo erosivo. O objetivo deste trabalho foi avaliar as condições hidráulicas e as relações de desagregação do solo e de resistência ao escoamento com a presença de resíduos vegetais na erosão em entressulcos. O experimento foi realizado no laboratório, com um Argissolo Vermelho distrófico típico, em parcelas com 0,10 m m-1 de declive sob chuva simulada. O solo foi coberto por resíduos vegetais de palha de soja, nas doses de 0, 0,05, 0,1, 0,2, 0,4 e 0,8 kg m-2. O aumento na cobertura do solo (CS com resíduos vegetais elevou a altura da lâmina de escoamento e a rugosidade hidráulica e reduziu a velocidade média do escoamento, provocada pelo aumento das forças viscosas promovida pela interposição física dos resíduos ao escoamento. O resultado é a redução na taxa de desagregação do solo (Di. A Di foi de 5,35x10-4 kg m-2 s-1 para solo descoberto e 1,50x10-5 kg m-2 s-1 em solo com 100% de cobertura na maior dose de palha. Os modelos de Laflen e potencial foram adequados para estimar o coeficiente de cobertura para resíduo em contato direto com a superfície do solo em função da cobertura do solo.Soil surface cover with crop residue modifies surface flow characteristics, generated by excess rainfall, and soil detachment and sediment transport resulting from the erosion process. The objective of this study was to evaluate the hydraulic conditions, detachment and flow resistance on interrill erosion on soil covered with residue. The experiment was conducted in the laboratory, on a Hapludult soil at a slope of 0.10 m m-1, under simulated rainfall and soil surface covered with soybean residue at the rates of 0, 0.05, 0.1, 0.2, 0.4, and 0.8 kg m-2. The increase in soil surface cover (SC with residue, caused an increase in water flow

  7. Ancestral Reconstruction of a Pre-LUCA Aminoacyl-tRNA Synthetase Ancestor Supports the Late Addition of Trp to the Genetic Code.

    Science.gov (United States)

    Fournier, G P; Alm, E J

    2015-04-01

    The genetic code was likely complete in its current form by the time of the last universal common ancestor (LUCA). Several scenarios have been proposed for explaining the code's pre-LUCA emergence and expansion, and the relative order of the appearance of amino acids used in translation. One co-evolutionary model of genetic code expansion proposes that at least some amino acids were added to the code by the ancient divergence of aminoacyl-tRNA synthetase (aaRS) families. Of all the amino acids used within the genetic code, Trp is most frequently claimed as a relatively recent addition. We observe that, since TrpRS and TyrRS are paralogous protein families retaining significant sequence similarity, the inferred sequence composition of their ancestor can be used to evaluate this co-evolutionary model of genetic code expansion. We show that ancestral sequence reconstructions of the pre-LUCA paralog ancestor of TyrRS and TrpRS have several sites containing Tyr, yet a complete absence of sites containing Trp. This is consistent with the paralog ancestor being specific for the utilization of Tyr, with Trp being a subsequent addition to the genetic code facilitated by a process of aaRS divergence and neofunctionalization. Only after this divergence could Trp be specifically encoded and incorporated into proteins, including the TyrRS and TrpRS descendant lineages themselves. This early absence of Trp is observed under both homogeneous and non-homogeneous models of ancestral sequence reconstruction. Simulations support that this observed absence of Trp is unlikely to be due to chance or model bias. These results support that the final stages of genetic code evolution occurred well within the "protein world," and that the presence-absence of Trp within conserved sites of ancient protein domains is a likely measure of their relative antiquity, permitting the relative timing of extremely early events within protein evolution before LUCA.

  8. Arabidopsis Indole Synthase,a Homolog of Tryptophan Synthase Alpha,is an Enzyme Involved in the Trp-independent Indole-containing Metabolite Biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Rui Zhang; Bing Wang; Jian Ouyang; Jiayang Li; Yonghong Wang

    2008-01-01

    The plant tryptophan (Trp) biosynthetic pathway produces many secondary metabolites with diverse functions.Indole-3-acetic acid (IAA),proposed as a derivative from Trp or its precursors,plays an essential role in plant growth and development.Although the Trp-dependant and Trp-independent IAA biosynthetic pathways have been proposed,the enzymes,reactions and regulatory mechanisms are largely unknown.In Arabidopsis,indole-3-glycerol phosphate (IGP) is suggested to serve as a branchpoint component in the Trp-independent IAA biosynthesis.To address whether other enzymes in addition to Trp synthase α(TSA1) catalyze IGP cleavage,we identified and characterized an indole synthase (INS) gene,a homolog of TSA1 in Arabidopsis.INS exhibits different subcellular localization from TSA1 owing to the lack of chloroplast transit peptide (cTP).In silico data show that the expression levels of INS and TSA1 in all examined organs are quite different.Histochemical staining of INS promoter-GUS transgenic lines indicates that INS is expressed in vascular tissue of cotyledons,hypocotyls,roots and rosette leaves as well as in flowers and siliques.INS is capable of complementing the Trp auxotrophy of Escherichia coil △trpA strain,which is defective in Trp synthesis due to the deletion of TSA.This implies that INS catalyzes the conversion of IGP to indole and may be involved in the biosynthesis of Trp-independent IAA or other secondary metabolites in Arabidopsis.

  9. Studies of the hyperthermophile Thermotoga maritima by random sequencing of cDNA and genomic libraries. Identification and sequencing of the trpEG (D) operon.

    Science.gov (United States)

    Kim, C W; Markiewicz, P; Lee, J J; Schierle, C F; Miller, J H

    1993-06-20

    Random sequencing of cDNA and genomic libraries has been used to study the genome of the hyperthermophile Thermotoga maritima. To date, 175 unique clones have been analyzed by comparing short sequence tags with known proteins in the PIR and GenBank databases. We find that a significant proportion of sequences can be matched to previously identified protein from non-Thermotoga sources. A high match rate was obtained from an oligo(dT)-primed cDNA library, where one-third of all unique sequences analyzed (21/65) shared high amino acid sequence similarity with proteins in the PIR and GenBank databases. Also, approximately one-third of the unique sequences from a second cDNA library (28/89), constructed with random oligo primers, could be matched to sequences in PIR and GenBank. Identification of genes from the oligo(dT)-primed cDNA library indicates that some Thermotoga mRNAs are polyadenylated. Genes have also been identified from a 1 to 2 kb genomic DNA library. Here, (3/21) of genomic sequences analyzed could be matched to protein in PIR and GenBank. One of the genomic clones had high sequence similarity to the tryptophan synthesis gene anthranilate synthase component I (trpE). Using this sequence tag, the Thermotoga trp operon was isolated and sequenced. The Thermotoga maritima trp operon is arranged with trpE forming an overlapping transcript with a second protein consisting of a fusion of anthranilate synthase component II (trpG) and anthranilate phosphoribosyltransferse (trpD). With regard to the fusion, the operon organization is similar to Escherichia coli and Salmonella typhimurium, but lacks the classic attenuation system of enteric bacteria. Amino acid sequence comparison with 19 trpE, 18 trpG and 14 trpD genes from other organisms suggest that the Thermotoga trp genes resemble corresponding genes from other thermophiles more closely than expected.

  10. Molecular basis of TRAP-5'SL RNA interaction in the Bacillus subtilis trp operon transcription attenuation mechanism.

    Science.gov (United States)

    McGraw, Adam P; Mokdad, Ali; Major, François; Bevilacqua, Philip C; Babitzke, Paul

    2009-01-01

    Expression of the Bacillus subtilis trpEDCFBA operon is regulated by the interaction of tryptophan-activated TRAP with 11 (G/U)AG trinucleotide repeats that lie in the leader region of the nascent trp transcript. Bound TRAP prevents folding of an antiterminator structure and favors formation of an overlapping intrinsic terminator hairpin upstream of the trp operon structural genes. A 5'-stem-loop (5'SL) structure that forms just upstream of the triplet repeat region increases the affinity of TRAP-trp RNA interaction, thereby increasing the efficiency of transcription termination. Single-stranded nucleotides in the internal loop and in the hairpin loop of the 5'SL are important for TRAP binding. We show here that altering the distance between these two loops suggests that G7, A8, and A9 from the internal loop and A19 and G20 from the hairpin loop constitute two structurally discrete TRAP-binding regions. Photochemical cross-linking experiments also show that the hairpin loop of the 5'SL is in close proximity to the flexible loop region of TRAP during TRAP-5'SL interaction. The dimensions of B. subtilis TRAP and of a three-dimensional model of the 5'SL generated using the MC-Sym and MC-Fold pipeline imply that the 5'SL binds the protein in an orientation where the helical axis of the 5'SL is perpendicular to the plane of TRAP. This interaction not only increases the affinity of TRAP-trp leader RNA interaction, but also orients the downstream triplet repeats for interaction with the 11 KKR motifs that lie on TRAP's perimeter, increasing the likelihood that TRAP will bind in time to promote termination.

  11. TrpA1 Regulates Defecation of Food-Borne Pathogens under the Control of the Duox Pathway.

    Directory of Open Access Journals (Sweden)

    Eun Jo Du

    2016-01-01

    Full Text Available Pathogen expulsion from the gut is an important defense strategy against infection, but little is known about how interaction between the intestinal microbiome and host immunity modulates defecation. In Drosophila melanogaster, dual oxidase (Duox kills pathogenic microbes by generating the microbicidal reactive oxygen species (ROS, hypochlorous acid (HOCl in response to bacterially excreted uracil. The physiological function of enzymatically generated HOCl in the gut is, however, unknown aside from its anti-microbial activity. Drosophila TRPA1 is an evolutionarily conserved receptor for reactive chemicals like HOCl, but a role for this molecule in mediating responses to gut microbial content has not been described. Here we identify a molecular mechanism through which bacteria-produced uracil facilitates pathogen-clearing defecation. Ingestion of uracil increases defecation frequency, requiring the Duox pathway and TrpA1. The TrpA1(A transcript spliced with exon10b (TrpA1(A10b that is present in a subset of midgut enteroendocrine cells (EECs is critical for uracil-dependent defecation. TRPA1(A10b heterologously expressed in Xenopus oocytes is an excellent HOCl receptor characterized with elevated sensitivity and fast activation kinetics of macroscopic HOCl-evoked currents compared to those of the alternative TRPA1(A10a isoform. Consistent with TrpA1's role in defecation, uracil-excreting Erwinia carotovora showed higher persistence in TrpA1-deficient guts. Taken together, our results propose that the uracil/Duox pathway promotes bacteria expulsion from the gut through the HOCl-sensitive receptor, TRPA1(A10b, thereby minimizing the chances that bacteria adapt to survive host defense systems.

  12. Heterogeneous nano-Fe/Ca/CaO catalytic ozonation for selective surface hydrophilization of plastics containing brominated and chlorinated flame retardants (B/CFRs): separation from automobile shredder residue by froth flotation.

    Science.gov (United States)

    Mallampati, Srinivasa Reddy; Lee, Byoung Ho; Mitoma, Yoshiharu; Simion, Cristian

    2016-12-09

    One method of weakening the inherently hydrophobic surface of plastics relevant to flotation separation is heterogeneous nano-Fe/Ca/CaO catalytic ozonation. Nano-Fe/Ca/CaO-catalyzed ozonation for 15 min efficiently decreases the surface hydrophobicity of brominated and chlorinated flame retardant (B/CFR)-containing plastics (such as acrylonitrile-butadienestyrene (ABS), high-impact polystyrene (HIPS), and polyvinyl chloride (PVC)) in automobile shredder residue (ASR) to such an extent that their flotation ability is entirely depressed. Such a hydrophilization treatment also stimulates the ABS, HIPS, and PVC surface roughness, wetting of the surface, and the thermodynamic equilibrium conditions at the surface and ultimately changes surface polarity. SEM-EDS, AFM, and XPS analyses of the PVC and ABS surfaces demonstrated a marked decrease in [Cl/Br] and a significant increase in the number of hydrophilic groups, such as C-O, C=O, and (C=O)-O. Under froth flotation conditions at 50 rpm, about 99.5 % of ABS and 99.5 % of HIPS in ASR samples settled out, resulting in a purity of 98 and 98.5 % for ABS and HIPS in ASR samples, respectively. Furthermore, at 150 rpm, we also obtained 100 % PVC separation in the settled fraction, with 98 % purity in ASR. Total recovery of non-B/CFR-containing plastics reached nearly 100 % in the floating fraction. The amount of nano-Fe/Ca/CaO reagent employed during ozonation is very small, and additional removal of surface contaminants from the recycled ASR plastic surfaces by ozonation makes the developed process simpler, greener, and more effective.

  13. Recoveries of trace pseudoephedrine and methamphetamine residues from impermeable household surfaces: implications for sampling methods used during remediation of clandestine methamphetamine laboratories.

    Science.gov (United States)

    Abdullah, A F Lim; Miskelly, Gordon M

    2010-04-15

    Evaluation of the risk posed by contaminants present during and after decontamination of clandestine methamphetamine laboratories requires a connection between the levels of contaminants measured and those actually present at the scene. The recoveries of pseudoephedrine and methamphetamine from glass, stainless steel, and a range of impermeable surfaces likely to be found in a clandestine laboratory were examined, using GC-MS of derivatized samples as the analytical method. When surfaces had been cleaned prior to drug deposition, wiping with water-dampened filter paper can recover 60-80% of pseudoephedrine immediately after deposition, and at least 50% of the pseudoephedrine still present on a surface after 2 days when deposited at a surface concentration of 2.5 microg/100 cm(2). Wiping with methanol-dampened filter paper could recover 60-90% of the methamphetamine immediately after deposition, and could recover at least 50-60% of the methamphetamine still present after 2 days when 0.6 microg/100 cm(2) was initially deposited on the surface. Recoveries were lower for surfaces that had not been pre-cleaned. Methamphetamine and pseudoephedrine showed significant volatility in both the free base and hydrochloride forms, with experiments in an enclosed format showing up to half the recovered drug being present on a glass plate held about 4mm above a substrate contaminated with one of the drugs at the above surface concentrations after 2 days. It is therefore important to remove any visible bulk contaminants and remove obvious pseudoephedrine or methamphetamine-contaminated surfaces prior to heating, ventilation or sealing of a clandestine laboratory to avoid redistribution of material around the site. A revised method for pseudoephedrine analysis was developed that could also detect the pseudoephedrine-formaldehyde adduct that can form from trace pseudoephedrine present at clandestine laboratories. (c) 2009 Elsevier B.V. All rights reserved.

  14. Application of residual stress measurement method in analysis of vault defect on sheet metal surface%残余应力测试法在分析薄板起拱缺陷中的应用

    Institute of Scientific and Technical Information of China (English)

    罗亚军; 杨建华; 何丹农; 张永清

    2001-01-01

    某公司在生产某钢号薄板(厚度大于0.35mm)时容易产生起拱现象,而导致产品报废。为了找出产生此缺陷的原因,通过应力释放法中的钻孔法对薄板表面的残余应力进行测试,结果发现薄板加工过程中的某工序使薄板表面产生较大的残余应力而且其分布也不均匀,这是导致薄板最终产生缺陷的主要原因。%Sheet metal often turns vaulted when its thickness is more than 0.35mm which happens in the production in one corporation, which makes the products unqualified. In order to find out the cause that leads to the default, residual stress on the surface of sheet metal after every working procedure is measured by the Hole-drilling Method. The results show that a certain working procedure makes much more residual stress and its distribution is asymmetric on sheet metal surface in sheet metal working process. That is the main reason to make sheet metal unqualified.

  15. 纳米结构陶瓷涂层磨削表面残余应力的实验设计%The Experiment Designing of the Surface Residual Stresses after Grinding for Nanostructured Ceramic Coatings

    Institute of Scientific and Technical Information of China (English)

    刘伟香; 王瑶

    2014-01-01

    The nanostructured tungsten carbide / cobalt ( n-WC/12Co ) ceramic coating materials are taken as grinding object , grinding machine and three kinds of grinding wheel are cleverly chosen , by using X ray diffraction instrument and the design of grinding parameters , the paper provided basis for nanostructured ceramic coatings grinding experiments and surface residual stress measurement and follow-up study "grinding parameters on the grinding surface residual stress influence rule".%以近年来新出现的纳米结构碳化钨/钴( n- WC/12Co )陶瓷涂层材料为磨削对象,巧妙选择磨削实验用磨床和三种砂轮,利用X射线衍射仪等设备,通过设计磨削参数,为纳米结构陶瓷涂层的磨削实验和表面残余应力的测量及后续研究“磨削参数对磨削表面残余应力的影响规律”提供依据。

  16. Simulation Research on the Surface Residual Stress of Ultrasonic Vibration Honing%超声振动珩磨表面残余应力数值模拟研究

    Institute of Scientific and Technical Information of China (English)

    郑华林; 张伟

    2015-01-01

    目的:磨削产生的残余应力对工件表面特性有重要影响,超声振动珩磨使磨粒具有极大的加速度而改变了材料去除机理,研究超声振动对工件表面残余应力的影响及产生机理。方法分析残余应力形成有限元理论,建立基于热弹塑性有限元法的超声振动单颗CBN磨粒切削40 CrNiMoA热力耦合有限元模型,并设置两次切削、卸载、约束转换及冷却等分析步。通过数值模拟得到不同振动参数下表面残余应力的分布情况,并对模拟结果进行分析。结果有限元计算得到的各分析阶段应力分布存在差别,超声振动参数设置达到仿真要求;对磨粒施加超声振动后珩磨力下降约26%,珩磨热降低约17%,切向残余压应力有所减小,垂直珩磨速度方向拉应力减小并向压应力转变。结论超声振动使珩磨力和珩磨热有一定程度降低从而改变了残余应力的分布及数值;振动频率在20 kHz波动时对残余应力的影响不大;磨削速度减小,切向残余压应力增大,垂直磨削方向残余拉应力减小;振幅增大时,切向残余压应力减小,垂直方向残余应力增大。%ABSTRACT:Objective To study the influence on residual stress as the grinding residual stress has important influence on work-piece surface characteristics and ultrasonic vibration honing makes the grains obtain a great acceleration which changes the material removal mechanism. Methods The residual stress finite element theory was analyzed, the coupled thermal-mechanical finite ele-ment model was established for the ultrasonic vibration honing of a single CBN grain cutting 40CrNiMoA based on the thermal elas-tic-plastic theory. Steps of two cutting processes, unloading, constraint transformation and the cooling analysis were set. The sur-face residual stress distribution was obtained by numerical simulation under different vibration parameters and the simulation results

  17. Systematic and quantitative mRNA expression analysis of TRP channel genes at the single trigeminal and dorsal root ganglion level in mouse

    Directory of Open Access Journals (Sweden)

    Vandewauw Ine

    2013-02-01

    Full Text Available Abstract Background Somatosensory nerve fibres arising from cell bodies within the trigeminal ganglia (TG in the head and from a string of dorsal root ganglia (DRG located lateral to the spinal cord convey endogenous and environmental stimuli to the central nervous system. Although several members of the transient receptor potential (TRP superfamily of cation channels have been implicated in somatosensation, the expression levels of TRP channel genes in the individual sensory ganglia have never been systematically studied. Results Here, we used quantitative real-time PCR to analyse and compare mRNA expression of all TRP channels in TG and individual DRGs from 27 anatomically defined segments of the spinal cord of the mouse. At the mRNA level, 17 of the 28 TRP channel genes, TRPA1, TRPC1, TRPC3, TRPC4, TRPC5, TRPM2, TRPM3, TRPM4, TRPM5, TRPM6, TRPM7, TRPM8, TRPV1, TRPV2, TRPV4, TRPML1 and TRPP2, were detectable in every tested ganglion. Notably, four TRP channels, TRPC4, TRPM4, TRPM8 and TRPV1, showed statistically significant variation in mRNA levels between DRGs from different segments, suggesting ganglion-specific regulation of TRP channel gene expression. These ganglion-to-ganglion differences in TRP channel transcript levels may contribute to the variability in sensory responses in functional studies. Conclusions We developed, compared and refined techniques to quantitatively analyse the relative mRNA expression of all TRP channel genes at the single ganglion level. This study also provides for the first time a comparative mRNA distribution profile in TG and DRG along the entire vertebral column for the mammalian TRP channel family.

  18. Resíduos vegetais na superfície do solo afetam a acidez do solo e a eficiência do herbicida flumetsulam Soil surface residues affect soil acidity and flumetsulam herbicid efficiency

    Directory of Open Access Journals (Sweden)

    Antonio Sergio do Amaral

    2000-10-01

    Full Text Available Com o objetivo de avaliar o potencial de resíduos de duas espécies vegetais em alterar a acidez do solo e a atividade do herbicida flumetsulam, foi conduzido um experimento em casa de vegetação no Departamento de Solos da Universidade Federal do Rio Grande do Sul, em novembro de 1998. Amostras da camada superficial (0-20cm de um cambissolo húmico distrófico foram colocadas em vasos, com capacidade de 1,0kg, onde se aplicou na superfície: 0, 5 e 10t ha-1 de resíduos de aveia-preta e de ervilhaca comum, com e sem aplicação do herbicida. Utilizou-se aveia-branca como cultura reagente. O delineamento experimental foi o inteiramente casualizado com três repetições. O resíduo da ervilhaca aumentou o pH do solo e ambos os resíduos diminuíram o alumínio trocável na camada de 0-3cm de profundidade, sendo o efeito da ervilhaca mais pronunciado. O decréscimo do alumínio trocável foi relacionado a dois mecanismos: complexação pela matéria orgânica e aumento do pH. Os resíduos vegetais aplicados sobre a superfície do solo aumentaram a eficiência do herbicida flumetsulam, especialmente quando o herbicida foi aplicado sobre os resíduos da leguminosa, que teve maior efeito na elevação do pH do solo.An experiment was conducted in the greenhouse of the Soils Department of the Federal University of Rio Grande do Sul, in November of 1998, to evaluate the potential of two crop residues to modify the acidity of the soil and the activity of the flumetsulam herbicide. The experimental units consisted of pots containing an Haplumbrept soil from the surface layer (0-20cm in with it was applyed 0, 5 and 10t ha-1 of black oat and of common vicia residues, with and without the herbicide application in the soil surface, and arranged in a complete randomized design, with three replications. White oat was used as test culture. Vicia residues increase soil pH and both residues decreased exchangeable Al in the 0-3cm layer, with the effect of vicia

  19. 钻环测量水泥混凝土板表面残余应力的试验研究%THE RESEARCH OF CORE RING METHOD TO MEASURE RESIDUAL STRESS ON THE SURFACE OF CEMENT CONCRETE SLAB

    Institute of Scientific and Technical Information of China (English)

    李新凯; 侯相深; 马松林

    2012-01-01

    水泥混凝土在强度形成过程中,水泥砂浆体积变形受到约束后将导致水泥混凝土内产生残余应力。对水泥混凝土路面板而言,板表层水泥砂浆的体积变形受外部环境影响更严重,因此板表面存在较大的残余应力,超过其抗弯拉强度时将导致微观裂缝的出现。目前,根据金属工程中测量表面残余应力的钻孔法,相关学者提出了钻环测量水泥混凝土表面残余应力的方法,并对其可行性进行了分析。该文将在该方法可行性分析的基础上,建立有限元模型对钻环过程进行模拟,确定室内试验时钻环法的相关技术参数,然后浇注水泥混凝土板,考虑加载和不加载两种工况进行室内试验,试验结果表明:钻环能够释放板表面的残余应力,该方法可以用于水泥混凝土表面残余应力测量。%In the procedure of concrete strength development, the changes in cementitious material volume results in the production of internal residual stress. Because the volume changes of cement pasted on the surface are influnced more obviously by environmental factors, the residual stress on the surface of rigid pavement slab will develop quickly and may lead to the appearance of micro crackings. According to the hole drilling strain gage (HDSG) method which is often used to measure residual stress in metal structure, some researchers developed the core ring strain gage (CRSG) method for concrete structures and the feasibility of this method was studied. In this paper a finite element model (FEM) is developed to simulate the process of core ring drilling in concrete slab. The parameters in CRSG method are selected based on FEM's results for different cases. Large scale slabs are casted in laboratory and tests using CRSG method to measure residual stress are conducted. The test results of two cases (loading on slab comer and no loading) indicate that CRSG method can release

  20. Spatial and temporal distribution of 13C labelled plant residues in soil aggregates and Lumbricus terrestris surface casts: A combination of Transmission Electron Microscopy and Nanoscale Secondary Ion Mass Spectrometry

    Science.gov (United States)

    Vidal, Alix; Remusat, Laurent; Watteau, Françoise; Derenne, Sylvie; Quenea, Katell

    2016-04-01

    Earthworms play a central role in litter decomposition, soil structuration and carbon cycling. They ingest both organic and mineral compounds which are mixed, complexed with mucus and dejected in form of casts at the soil surface and along burrows. Bulk isotopic or biochemical technics have often been used to study the incorporation of litter in soil and casts, but they could not reflect the complex interaction between soil, plant and microorganisms at the microscale. However, the heterogeneous distribution of organic carbon in soil structures induces contrasted microbial activity areas. Nano-scale secondary ion mass spectrometry (NanoSIMS), which is a high spatial resolution method providing elemental and isotopic maps of organic and mineral materials, has recently been applied in soil science (Herrmann et al., 2007; Vogel et al., 2014). The combination of Nano-scale secondary ion mass spectrometry (NanoSIMS) and Transmission Electron Microscopy (TEM) has proven its potential to investigate labelled residues incorporation in earthworm casts (Vidal et al., 2016). In line of this work, we studied the spatial and temporal distribution of plant residues in soil aggregates and earthworm surface casts. This study aimed to (1) identify the decomposition states of labelled plant residues incorporated at different time steps, in casts and soil, (2) identify the microorganisms implied in this decomposition (3) relate the organic matter states of decomposition with their 13C signature. A one year mesocosm experiment was set up to follow the incorporation of 13C labelled Ryegrass (Lolium multiflorum) litter in a soil in the presence of anecic earthworms (Lumbricus terrestris). Soil and surface cast samples were collected after 8 and 54 weeks, embedded in epoxy resin and cut into ultra-thin sections. Soil was fractionated and all and analyzed with TEM and NanoSIMS, obtaining secondary ion images of 12C, 16O, 12C14N, 13C14N and 28Si. The δ13C maps were obtained using the 13C14

  1. Synthesis of gastrin antagonists, analogues of the C-terminal tetrapeptide of gastrin, by introduction of a beta-homo residue.

    Science.gov (United States)

    Rodriguez, M; Fulcrand, P; Laur, J; Aumelas, A; Bali, J P; Martinez, J

    1989-03-01

    A series of analogues of Boc-Trp-Leu-Asp-Phe-NH2, a potent gastrin agonist, were synthesized by introducing a beta-homo residue in the sequence. These compounds were tested in vivo on acid secretion, in the anesthetized rat, and for their ability to inhibit binding of labeled gastrin to its receptors on gastric mucosal cells. These analogues behaved as gastrin antagonists. The most potent compounds in this series were Boc-Trp-Leu-beta-homo-Asp-NHCH2C6H5 (10) (IC50 = 1 microM, ED50 = 0.2 mg/kg), Boc-Trp-Leu-beta-homo-Asp-NHCH2CH2C6H5 (11) (IC50 = 0.75 microM, ED50 = 0.5 mg/kg), Boc-Trp-Leu-beta-homo-Asp-Phe-NH2 (12) (IC50 = 1.5 microM, ED50 = 0.1 mg/kg), and Boc-Trp-Leu-beta-homo-Asp-D-Phe-NH2 (13) (IC50 = 2 microM, ED50 = 0.1 mg/kg). We could demonstrate the importance of the region of the peptide bond between leucine and aspartic acid and of the structure of the C-terminal dipeptide Asp-Phe-NH2, for exhibiting biological activity on acid secretion.

  2. Environmental Temperature Differentially Modulates C. elegans Longevity through a Thermosensitive TRP Channel

    Directory of Open Access Journals (Sweden)

    Bi Zhang

    2015-06-01

    Full Text Available Temperature profoundly affects aging in both poikilotherms and homeotherms. A general belief is that lower temperatures extend lifespan, whereas higher temperatures shorten it. Although this “temperature law” is widely accepted, it has not been extensively tested. Here, we systematically evaluated the role of temperature in lifespan regulation in C. elegans. We found that, although exposure to low temperatures at the adult stage prolongs lifespan, low-temperature treatment at the larval stage surprisingly reduces lifespan. Interestingly, this differential effect of temperature on longevity in larvae and adults is mediated by the same thermosensitive TRP channel TRPA-1 that signals to the transcription factor DAF-16/FOXO. DAF-16/FOXO and TRPA-1 act in larva to shorten lifespan but extend lifespan in adulthood. DAF-16/FOXO differentially regulates gene expression in larva and adult in a temperature-dependent manner. Our results uncover complexity underlying temperature modulation of longevity, demonstrating that temperature differentially regulates lifespan at different stages of life.

  3. A TRP channel in the lysosome regulates large particle phagocytosis via focal exocytosis.

    Science.gov (United States)

    Samie, Mohammad; Wang, Xiang; Zhang, Xiaoli; Goschka, Andrew; Li, Xinran; Cheng, Xiping; Gregg, Evan; Azar, Marlene; Zhuo, Yue; Garrity, Abigail G; Gao, Qiong; Slaugenhaupt, Susan; Pickel, Jim; Zolov, Sergey N; Weisman, Lois S; Lenk, Guy M; Titus, Steve; Bryant-Genevier, Marthe; Southall, Noel; Juan, Marugan; Ferrer, Marc; Xu, Haoxing

    2013-09-16

    Phagocytosis of large extracellular particles such as apoptotic bodies requires delivery of the intracellular endosomal and lysosomal membranes to form plasmalemmal pseudopods. Here, we identified mucolipin TRP channel 1 (TRPML1) as the key lysosomal Ca2+ channel regulating focal exocytosis and phagosome biogenesis. Both particle ingestion and lysosomal exocytosis are inhibited by synthetic TRPML1 blockers and are defective in macrophages isolated from TRPML1 knockout mice. Furthermore, TRPML1 overexpression and TRPML1 agonists facilitate both lysosomal exocytosis and particle uptake. Using time-lapse confocal imaging and direct patch clamping of phagosomal membranes, we found that particle binding induces lysosomal PI(3,5)P2 elevation to trigger TRPML1-mediated lysosomal Ca2+ release specifically at the site of uptake, rapidly delivering TRPML1-resident lysosomal membranes to nascent phagosomes via lysosomal exocytosis. Thus phagocytic ingestion of large particles activates a phosphoinositide- and Ca2+-dependent exocytosis pathway to provide membranes necessary for pseudopod extension, leading to clearance of senescent and apoptotic cells in vivo.

  4. TRP channels: a missing bond in the entrainment mechanism of peripheral clocks throughout evolution.

    Science.gov (United States)

    Poletini, Maristela O; Moraes, Maria Nathália; Ramos, Bruno César; Jerônimo, Rodrigo; Castrucci, Ana Maria de Lauro

    2015-01-01

    Circadian rhythm may be understood as a temporal organization that works to orchestrate physiological processes and behavior in a period of approximately 24 h. Because such temporal organization has evolved in the presence of predictable environmental clues, such as day length, tides, seasons, and temperature, the organism has confronted the natural selection in highly precise intervals of opportunities and risks, generating temporal programs and resetting mechanisms, which are well conserved among different taxa of animals. The present review brings some evidence of how these programs may have co-evolved in systems able to deal with 2 or more environmental clues, and how they similarly function in different group of animals, stressing how important temperature and light were to establish the temporal organizations. For example, melanopsin and rhodopsin, photopigments present respectively in circadian and visual photoreceptors, are required for temperature discrimination in Drosophila melanogaster. These pigments may signal light and temperature via activation of cationic membrane channel, named transient-receptor potential channel (TRP). In fact, TRPs have been suggested to function as thermal sensor for various groups of animals. Another example is the clock machinery at the molecular level. A set of very-well conserved proteins, known as clock proteins, function as transcription factors in positive and negative auto-regulatory loops generating circadian changes of their expression, and of clock-controlled genes. Similar molecular machinery is present in organisms as diverse as cyanobacteria (Synechococcus), fungi (Neurospora), insects (Drosophila), and vertebrates including humans.

  5. Residual stress calculation method for fused silica polishing surface%熔石英元件抛光加工表面残余应力的计算方法

    Institute of Scientific and Technical Information of China (English)

    王洪祥; 侯晶; 严志龙; 朱本温; 陈贤华

    2015-01-01

    Fused silica glass is non⁃crystalline material, the traditional detection methods have complicated steps and low measurement accuracy, residual stresses are often unable to direct quantitative detection. A new residual stress calculation method for fused silica polishing surface was proposed based on brittle solid fracture mechanics, in which a series of nano⁃indentation experiments were conducted by a sharp indenter, and the sensitive parameters were extracted during indentation process, the experimental data were linear fit and the line slopes were determined, so residual stress was often calculated indirectly by measuring the other physical parameters change caused by residual stress. Comparative analysis showed that the calculation results had in good consistency with the test results obtained by stress birefringence analyzer, so the correctness of the residual stress calculation method was verified.%为解决传统检测方法无法直接定量检测非晶体熔石英玻璃表面残余应力的问题,基于脆性固体断裂力学理论,推导残余应力的理论计算公式,提出光学元件抛光加工表面残余应力计算新方法。采用尖锐压头进行纳米印压实验,提取压痕过程中对残余应力敏感的参数,并对实验数据进行线性拟合,确定拟合线的斜率,通过测量残余应力引起其他物理参数的变化计算残余应力。对比分析结果表明,计算得到残余应力值与应力双折射仪检测得到的数据基本吻合,验证了提出残余应力计算方法的正确性。

  6. Trp[superscript 2313]-His[superscript 2315] of Factor VIII C2 Domain Is Involved in Membrane Binding Structure of a Complex Between the C[subscript 2] Domain and an Inhibitor of Membrane Binding

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Zhuo; Lin, Lin; Yuan, Cai; Nicolaes, Gerry A.F.; Chen, Liqing; Meehan, Edward J.; Furie, Bruce; Furie, Barbara; Huang, Mingdong (Harvard-Med); (UAH); (Maastricht); (Chinese Aca. Sci.)

    2010-11-03

    Factor VIII (FVIII) plays a critical role in blood coagulation by forming the tenase complex with factor IXa and calcium ions on a membrane surface containing negatively charged phospholipids. The tenase complex activates factor X during blood coagulation. The carboxyl-terminal C2 domain of FVIII is the main membrane-binding and von Willebrand factor-binding region of the protein. Mutations of FVIII cause hemophilia A, whereas elevation of FVIII activity is a risk factor for thromboembolic diseases. The C2 domain-membrane interaction has been proposed as a target of intervention for regulation of blood coagulation. A number of molecules that interrupt FVIII or factor V (FV) binding to cell membranes have been identified through high throughput screening or structure-based design. We report crystal structures of the FVIII C2 domain under three new crystallization conditions, and a high resolution (1.15 {angstrom}) crystal structure of the FVIII C2 domain bound to a small molecular inhibitor. The latter structure shows that the inhibitor binds to the surface of an exposed {beta}-strand of the C2 domain, Trp{sup 2313}-His{sup 2315}. This result indicates that the Trp{sup 2313}-His{sup 2315} segment is an important constituent of the membrane-binding motif and provides a model to understand the molecular mechanism of the C2 domain membrane interaction.

  7. Assessment of planetary boundary layer and residual layer heights in the Northeastern U.S. using Lidar, a network of surface observations, and the WRF-STILT model

    Science.gov (United States)

    Barrera, Y.; Nehrkorn, T.; Hegarty, J. D.; Wofsy, S. C.; Gottlieb, E.; Sargent, M. R.; Decola, P.; Jones, T.

    2015-12-01

    Simulation of the planetary boundary layer (PBL) and residual layer (RL) are key requirements for forecasting air quality in cities and detecting transboundary air pollution events. This study combines information from a network of Mini Micropulse Lidar (MPL) instruments, the CALIOP satellite, meteorological and air pollution measuring sensors, and a particle-transport model to critically test mesoscale transport models at the regional level. Aerosol backscattering measurements were continuously taken with MPL units in various locations within the Northeastern U.S., between September 2012 to August 2015. Data is analyzed using wavelet covariance transforms and image processing techniques. Initial results for the city of Boston show a PBL growth rate between approx. 150 and 300 meters per hour, in the morning to early afternoon (~12-19 UTC). The RL was present throughout the night and day at approx. 1.3 to 2.0 km. Transboundary air pollution events were detected and quantified, and variations in concentrations of greenhouse gases and aerosols were also evaluated. Results were compared to information retrieved from Weather and Research Forecasting (WRF) model and the Stochastic Time-Inverted Lagrangian Transport (STILT) model.

  8. Residues of organophosphate pesticides used in vegetable cultivation in ambient air, surface water and soil in Bueng Niam Subdistrict, Khon Kaen, Thailand.

    Science.gov (United States)

    Harnpicharnchai, Kallaya; Chaiear, Naesinee; Charerntanyarak, Lertchai

    2013-11-01

    Agricultural pesticide utilization is one of the important problems in rural and urban crop-cultivated areas, with the majority of pollutants dispersing via ambient air, water and other natural pathways. This study was therefore conducted in a specially selected village which is known to be a leading vegetable growing area in Khon Kaen Province. The aim of the study was to assess pesticide residues, and measure the seasonal fluctuations in organophosphate concentrations during 2010 in the environment of a risk area. Samples from selected sites were collected in two phases: Phase I was in summer (during March to May) and Phase II was in winter (during October to December). A total of 150 samples were analyzed using gas chromatography with flame photometric detection. The results showed that dicrotophos, chlorpyrifos, profenofos and ethion were found at the highest concentrations in soil and at the lowest concentrations in ambient air (pwater samples, the highest mean concentration of a pesticide was 1.3757 +/- 0.5014 mg/l for dicrotophos in summer and 0.3629 +/- 0.4338 mg/l for ethion in winter. The highest mean concentration of a pesticide in soil samples was 42.2893 +/- 39.0711 mg/kg ethion in summer and 90.0000 +/- 24.1644 mg/kg of ethion in winter.

  9. Hepatic fibrinogen storage disease due to the fibrinogen γ375 Arg → Trp mutation "fibrinogen aguadilla" is present in Arabs

    Directory of Open Access Journals (Sweden)

    Abdulrahman Al-Hussaini

    2014-01-01

    Full Text Available The mutation γ375Arg → Trp (fibrinogen Aguadilla is one of four mutations (Brescia, Aguadilla, Angers, and AI duPont capable of causing hepatic storage of fibrinogen. It has been observed in four children from the Caribbean, Europe, and Japan, suffering from cryptogenic liver disease. We report the first case of hepatic fibrinogen storage disease in Arabs due to a mutation in the fibrinogen γ-chain gene in a 3-year-old Syrian girl presenting with elevated liver enzymes. The finding of an impressive accumulation of fibrinogen in liver cells raised the suspicion of endoplasmic reticulum storage disease. Sequencing of the fibrinogen genes revealed a γ375Arg → Trp mutation (fibrinogen Aguadilla in the child and in her father. In conclusion, when confronted with chronic hepatitis of unknown origin, one should check the plasma fibrinogen level and look carefully for the presence of hepatocellular intracytoplasmic globular inclusions to exclude hepatic fibrinogen storage disease.

  10. Identification and Molecular Characterization of a Gene Encoding a Protective Leishmania amazonensis Trp-Asp (WD) Protein

    OpenAIRE

    2004-01-01

    Several Leishmania proteins have been identified and characterized in pursuit of understanding pathogenesis and protection in cutaneous leishmaniasis. In the present study, we utilized sera from infected BALB/c mice to screen a Leishmania amazonensis amastigote cDNA expression library and obtained the full-length gene that encodes a novel Trp-Asp (WD) protein designated LAWD (for Leishmania antigenic WD protein). The WD family of proteins mediates protein-protein interactions and coordinates ...

  11. Análisis preliminar de la estructura primaria y secundaria del ARNtTrp en tortugas marinas

    Directory of Open Access Journals (Sweden)

    Harvey Infante-Rojas

    2015-06-01

    Full Text Available Actualmente existen siete especies de tortugas marinas, todas amenazadas o en riesgo inminente de extinción. Los estudios con ADN mitocondrial han permitido hacer acercamientos sobre filogenia, evolución, rutas migratorias y centros de dispersión, además para la identificación de polimorfismos y haplotipos, siendo base para planes de manejo y conservación. El presente estudio representa la primera descripción comparada de la estructura primaria y secundaria del ARNtTrp mitocondrial en tortugas marinas. Se realizó un alineamiento múltiple de 26 secuencias del gen que codifica para el ARNtTrp y se propuso la estructura secundaria utilizando el programa ARWEN. Se identificaron potenciales interacciones terciarias por homología comparada con el ARNtTrp de mamíferos. Los resultados mostraron una secuencia consenso de 76 bases con siete regiones conservadas que representan el 76 % de la molécula. Se identificaron polimorfismos que representan tres haplotipos para C. caretta, dos para C. mydas y uno para cada una de las demás especies. Las estructuras secundarias mostraron cambios nucleotídicos puntuales para cada especie y también mostraron que el tallo aceptor, el brazo TψC y el bucle anticodón son motivos conservados en el ARNtTrp de las tortugas marinas. Se encontró un enlace no canónico tipo A-A en el tallo DHU que podría considerarse característico de tortugas marinas. Además, se obtuvo una estructura secundaria consenso en donde se identificaron las siete regiones conservadas, seis posibles interacciones terciarias y el bucle DHU como región variable.

  12. Induction of therapeutic hypothermia by pharmacological modulation of temperature-sensitive TRP channels: theoretical framework and practical considerations.

    Science.gov (United States)

    Feketa, Viktor V; Marrelli, Sean P

    2015-01-01

    Therapeutic hypothermia has emerged as a remarkably effective method of neuroprotection from ischemia and is being increasingly used in clinics. Accordingly, it is also a subject of considerable attention from a basic scientific research perspective. One of the fundamental problems, with which current studies are concerned, is the optimal method of inducing hypothermia. This review seeks to provide a broad theoretical framework for approaching this problem, and to discuss how a novel promising strategy of pharmacological modulation of the thermosensitive ion channels fits into this framework. Various physical, anatomical, physiological and molecular aspects of thermoregulation, which provide the foundation for this text, have been comprehensively reviewed and will not be discussed exhaustively here. Instead, the first part of the current review, which may be helpful for a broader readership outside of thermoregulation research, will build on this existing knowledge to outline possible opportunities and research directions aimed at controlling body temperature. The second part, aimed at a more specialist audience, will highlight the conceptual advantages and practical limitations of novel molecular agents targeting thermosensitive Transient Receptor Potential (TRP) channels in achieving this goal. Two particularly promising members of this channel family, namely TRP melastatin 8 (TRPM8) and TRP vanilloid 1 (TRPV1), will be discussed in greater detail.

  13. High-Temperature unfolding of a trp-Cage mini-protein: a molecular dynamics simulation study

    Directory of Open Access Journals (Sweden)

    Seshasayee Aswin Sai Narain

    2005-03-01

    Full Text Available Abstract Background Trp cage is a recently-constructed fast-folding miniprotein. It consists of a short helix, a 3,10 helix and a C-terminal poly-proline that packs against a Trp in the alpha helix. It is known to fold within 4 ns. Results High-temperature unfolding molecular dynamics simulations of the Trp cage miniprotein have been carried out in explicit water using the OPLS-AA force-field incorporated in the program GROMACS. The radius of gyration (Rg and Root Mean Square Deviation (RMSD have been used as order parameters to follow the unfolding process. Distributions of Rg were used to identify ensembles. Conclusion Three ensembles could be identified. While the native-state ensemble shows an Rg distribution that is slightly skewed, the second ensemble, which is presumably the Transition State Ensemble (TSE, shows an excellent fit. The denatured ensemble shows large fluctuations, but a Gaussian curve could be fitted. This means that the unfolding process is two-state. Representative structures from each of these ensembles are presented here.

  14. Insulin-producing cells regulate the sexual receptivity through the painless TRP channel in Drosophila virgin females.

    Directory of Open Access Journals (Sweden)

    Takaomi Sakai

    Full Text Available In a variety of animal species, females hold a leading position in evaluating potential mating partners. The decision of virgin females to accept or reject a courting male is one of the most critical steps for mating success. In the fruitfly Drosophila melanogaster, however, the molecular and neuronal mechanisms underlying female receptivity are still poorly understood, particularly for virgin females. The Drosophila painless (pain gene encodes a transient receptor potential (TRP ion channel. We previously demonstrated that mutations in pain significantly enhance the sexual receptivity of virgin females and that pain expression in pain(GAL4 -positive neurons is necessary and sufficient for pain-mediated regulation of the virgin receptivity. Among the pain(GAL4 -positive neurons in the adult female brain, here we have found that insulin-producing cells (IPCs, a neuronal subset in the pars intercerebralis, are essential in virgin females for the regulation of sexual receptivity through Pain TRP channels. IPC-specific knockdown of pain expression or IPC ablation strongly enhanced female sexual receptivity as was observed in pain mutant females. When pain expression or neuronal activity was conditionally suppressed in adult IPCs, female sexual receptivity was similarly enhanced. Furthermore, both pain mutations and the conditional knockdown of pain expression in IPCs depressed female rejection behaviors toward courting males. Taken together, our results indicate that the Pain TRP channel in IPCs plays an important role in controlling the sexual receptivity of Drosophila virgin females by positively regulating female rejection behaviors during courtship.

  15. Measurment Of Residual Stress In Ferromagnetic Materials

    Science.gov (United States)

    Namkung, Min; Yost, William T.; Kushnick, Peter W.; Grainger, John L.

    1992-01-01

    Magnetoacoustic (MAC) and magnetoacoustic emission (MAE) techniques combined to provide complete characterization of residual stresses in ferromagnetic structural materials. Combination of MAC and MAE techniques makes it possible to characterize residual tension and compression without being limited by surface conditions and unavailability of calibration standards. Significant in field of characterization of materials as well as detection of fatigue failure.

  16. Measurment Of Residual Stress In Ferromagnetic Materials

    Science.gov (United States)

    Namkung, Min; Yost, William T.; Kushnick, Peter W.; Grainger, John L.

    1992-01-01

    Magnetoacoustic (MAC) and magnetoacoustic emission (MAE) techniques combined to provide complete characterization of residual stresses in ferromagnetic structural materials. Combination of MAC and MAE techniques makes it possible to characterize residual tension and compression without being limited by surface conditions and unavailability of calibration standards. Significant in field of characterization of materials as well as detection of fatigue failure.

  17. Residual contact restraints in cryogenics

    Science.gov (United States)

    Cretegny, J. F.; Demonicault, J. M.

    The use of residual stress measurements to evaluate the state of cryogenic turbomachines, whose surfaces are worn by the working conductions in dry contact, is addressed. Their contribution to the understanding of the reasons of possible ruptures is considered. It is stated that residual stress measurements should be used as a complementary tool rather than as input data for models. It is shown, thanks to two examples concerning the ball bearings and splines of the liquid hydrogen turbopump of the Vulcain engine, what can be expected from such techniques. Total exploitation of the results has still to be done, but preliminary results are quite encouraging.

  18. Strong Enrichment of Aromatic Residues in Binding Sites from a Charge-neutralized Hyperthermostable Sso7d Scaffold Library*

    Science.gov (United States)

    Kiefer, Jonathan D.; Srinivas, Raja R.; Lobner, Elisabeth; Tisdale, Alison W.; Mehta, Naveen K.; Yang, Nicole J.; Tidor, Bruce; Wittrup, K. Dane

    2016-01-01

    The Sso7d protein from the hyperthermophilic archaeon Sulfolobus solfataricus is an attractive binding scaffold because of its small size (7 kDa), high thermal stability (Tm of 98 °C), and absence of cysteines and glycosylation sites. However, as a DNA-binding protein, Sso7d is highly positively charged, introducing a strong specificity constraint for binding epitopes and leading to nonspecific interaction with mammalian cell membranes. In the present study, we report charge-neutralized variants of Sso7d that maintain high thermal stability. Yeast-displayed libraries that were based on this reduced charge Sso7d (rcSso7d) scaffold yielded binders with low nanomolar affinities against mouse serum albumin and several epitopes on human epidermal growth factor receptor. Importantly, starting from a charge-neutralized scaffold facilitated evolutionary adaptation of binders to differentially charged epitopes on mouse serum albumin and human epidermal growth factor receptor, respectively. Interestingly, the distribution of amino acids in the small and rigid binding surface of enriched rcSso7d-based binders is very different from that generally found in more flexible antibody complementarity-determining region loops but resembles the composition of antibody-binding energetic hot spots. Particularly striking was a strong enrichment of the aromatic residues Trp, Tyr, and Phe in rcSso7d-based binders. This suggests that the rigidity and small size of this scaffold determines the unusual amino acid composition of its binding sites, mimicking the energetic core of antibody paratopes. Despite the high frequency of aromatic residues, these rcSso7d-based binders are highly expressed, thermostable, and monomeric, suggesting that the hyperstability of the starting scaffold and the rigidness of the binding surface confer a high tolerance to mutation. PMID:27582495

  19. Energy expenditure, body composition and insulin response to glucose in male twins discordant for the Trp64Arg polymorphism of the beta3-adrenergic receptor gene

    DEFF Research Database (Denmark)

    Højlund, K; Christiansen, C; Bjørnsbo, K S;

    2006-01-01

    and environmental background, the Trp64Arg polymorphism of the beta3AR gene is associated with lower fat mass, fasting insulin levels and an appropriate insulin response to glucose. Thus, heterozygosity for the Trp64Arg variant is unlikely to increase the risk of obesity, insulin resistance or type 2 diabetes.......AIM: The tryptophan to arginine change in position 64 (Trp64Arg) polymorphism of the beta3-adrenergic receptor (beta3AR) gene has been associated with an increased prevalence of obesity, insulin resistance and type 2 diabetes. In this, decreased rates of energy expenditure and impaired insulin......-ray absorptiometry scanning and energy expenditure by indirect and direct calorimetry. RESULTS: Twins heterozygous for the Trp64Arg polymorphism showed significantly lower fat mass independent of the method used, and significantly lower fasting insulin and glucose concentrations compared with their homozygous wild...

  20. Association of the beta3-adrenergic receptor Trp64Arg polymorphism with common metabolic traits: studies of 7605 middle-aged white people

    DEFF Research Database (Denmark)

    Gjesing, A. P.; Andersen, G; Borch-Johnsen, K;

    2008-01-01

    of type 2 diabetes and obesity in a relatively large, homogenous study population. METHODS: The Trp64Arg polymorphism was genotyped in 7605 Danish subjects using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Association was examined in case-control studies of obesity (1529...... cases and 6049 controls) and type 2 diabetes (1373 cases and 4742 controls) and quantitative trait analyses among 5822 individuals. Furthermore, the association of Trp64Arg with type 2 diabetes was examined in a meta-analysis. RESULTS: The Trp64Arg polymorphism was not associated with obesity. However...... (p=0.02). After sex stratification these associations were only present among women. Furthermore, the Arg-allele was borderline associated with type 2 diabetes in a meta-analysis of the present and 26 previous studies (p=0.06, OR1.27 (CI: 0.99-1.63)) (n=18891). CONCLUSION/INTERPRETATION: Trp64Arg...

  1. Characterization of TRAP-mediated regulation of the B. subtilis trp operon using in vitro transcription and transcriptional reporter fusions in vivo.

    Science.gov (United States)

    McAdams, Natalie M; Gollnick, Paul

    2015-01-01

    In Bacillus subtilis, transcription of the tryptophan biosynthetic operon is regulated by an attenuation mechanism involving two alternative RNA secondary structures in the 5' leader region upstream of the structural genes. Regulation is accomplished, at least in part, by controlling which RNA structure forms during transcription of the operon. When intracellular tryptophan levels are high, the trp RNA-binding attenuation protein (TRAP) binds to the nascent trp mRNA to promote formation of a transcription terminator structure so as to induce transcription termination prior to the structural genes. In limiting tryptophan, TRAP does not bind, the alternative antiterminator RNA structure forms, and the operon is transcribed. Several in vitro and in vivo assays have been utilized to study TRAP-mediated regulation of both transcription and translation. Here, we describe using in vitro transcription attenuation assays and in vivo trp-lacZ fusions to examine TRAP-mediated regulation of the trp genes.

  2. Food Compounds Activating Thermosensitive TRP Channels in Asian Herbal and Medicinal Foods.

    Science.gov (United States)

    Watanabe, Tatsuo; Terada, Yuko

    2015-01-01

    There are several thermosensitive transient receptor potential (TRP) ion channels including capsaicin receptor, TRPV1. Food components activating TRPV1 inhibit body fat deposition through sympathetic nerve stimulation. TRPA1 is another pungency sensor for pungent compounds and is mainly coexpressed with TRPV1 in sensory nerve endings. Therefore, TRPA1 activation is expected to have an anti-obesity effect similar to TRPV1 activation. We have searched for agonists for TRPV1 and TRPA1 in vitro from Asian spices by the use of TRPV1- and TRPA1-expressing cells. Further, we performed food component addition tests to high-fat and high-sucrose diets in mice. We found capsiate, capsiconiate, capsainol from hot and sweet peppers, several piperine analogs from black pepper, gingeriols and shogaols from ginger, and sanshools and hydroxysanshools from sansho (Japanese pepper) to be TRPV1 agonists. We also identified several sulfides from garlic and durian, hydroxy fatty acids from royal jelly, miogadial and miogatrial from mioga (Zingiber mioga), piperine analogs from black pepper, and acetoxychavicol acetate (ACA) from galangal (Alpinia galanga) as TRPA1 agonists. Piperine addition to diets diminished visceral fats and increased the uncoupling protein 1 (UCP1) in interscapular brown adipose tissue (IBAT), and black pepper extract showed stronger effects than piperine. Cinnamaldehyde and ACA as TRPA1 agonists inhibited fat deposition and increased UCP1. We found that several agonists of TRPV1 and TRPA1 and some agonists of TRPV1 and TRPA1 inhibit visceral fat deposition in mice. The effects of such compounds on humans remain to be clarified, but we expect that they will be helpful in the prevention of obesity.

  3. Bioaccessible Porosity: A new approach to assess residual contamination after bioremediation of hydrophobic organic compounds in sub-surface microporous environments

    Science.gov (United States)

    Akbari, A.; Ghoshal, S.

    2016-12-01

    We define a new parameter, "bioaccessible porosity", the fraction of aggregate volume accessible to soil bacteria, towards a priori assessment of hydrocarbon bioremediation end points. Microbial uptake of poorly soluble hydrocarbons occurs through direct uptake or micellar solubilzation/emulsification associated with biosurfactant production, and requires close proximity of bacteria and hydrocarbon phase. In subsurface microporous environments, bioremediation rates are attenuated when residual hydrophobic contamination is entrapped in sterically restrictive environments which is not accessible to soil bacteria. This study presents new approaches for characterization of the microstructure of porous media and as well, the ability of indigenous hydrocarbon degraders to access to a range of pore sizes. Bacterial access to poorly soluble hydrocarbons in soil micro pores were simulated with bioreactors with membranes with different pore sizes containing the hydrocarbon degrading bacteria, Dietzia maris. D. maris is Gram-positive, and nonmotile that we isolated as the major hydrocarbon degrader from a fine-grained, weathered, hydrocarbon-contaminated site soil. Under nutritional stress, planktonic D. maris cells were aggregated and accessed 5 µm but not 3 µm and smaller pores. However, when hexadecane was available at the pore mouth, D. maris colonized the pore mouth, and accessed pores as small as 0.4 µm. This suggests bacterial accessibility to different pore sizes is regulated by nutritional conditions. A combination of X-ray micro-CT scanning, gas adsorption and mercury intrusion porosimetry was used to characterize the range of pore sizes of soil aggregates. In case of the studied contaminated soil, the bioaccessible porosity were determined as 25% , 27% and 29% (assuming 4, 1, 0.4 µm respectively as accessibility criteria), and about 2.7% of aggregate volume was attributed to 0.006-0.4 µm pores. The 2% aggregate volume at an assumed saturation of 10% could

  4. The tumor suppressor gene Trp53 protects the mouse lens against posterior subcapsular cataracts and the BMP receptor Acvr1 acts as a tumor suppressor in the lens

    Directory of Open Access Journals (Sweden)

    Luke A. Wiley

    2011-07-01

    We previously found that lenses lacking the Acvr1 gene, which encodes a bone morphogenetic protein (BMP receptor, had abnormal proliferation and cell death in epithelial and cortical fiber cells. We tested whether the tumor suppressor protein p53 (encoded by Trp53 affected this phenotype. Acvr1 conditional knockout (Acvr1CKO mouse fiber cells had increased numbers of nuclei that stained for p53 phosphorylated on serine 15, an indicator of p53 stabilization and activation. Deletion of Trp53 rescued the Acvr1CKO cell death phenotype in embryos and reduced Acvr1-dependent apoptosis in postnatal lenses. However, deletion of Trp53 alone increased the number of fiber cells that failed to withdraw from the cell cycle. Trp53CKO and Acvr1;Trp53DCKO (double conditional knockout, but not Acvr1CKO, lenses developed abnormal collections of cells at the posterior of the lens that resembled posterior subcapsular cataracts. Cells from human posterior subcapsular cataracts had morphological and molecular characteristics similar to the cells at the posterior of mouse lenses lacking Trp53. In Trp53CKO lenses, cells in the posterior plaques did not proliferate but, in Acvr1;Trp53DCKO lenses, many cells in the posterior plaques continued to proliferate, eventually forming vascularized tumor-like masses at the posterior of the lens. We conclude that p53 protects the lens against posterior subcapsular cataract formation by suppressing the proliferation of fiber cells and promoting the death of any fiber cells that enter the cell cycle. Acvr1 acts as a tumor suppressor in the lens. Enhancing p53 function in the lens could contribute to the prevention of steroid- and radiation-induced posterior subcapsular cataracts.

  5. Computing Decoupled Residuals for Compact Disc Players

    DEFF Research Database (Denmark)

    Odgaard, Peter Fogh; Stoustrup, Jakob; Andersen, Palle

    2006-01-01

    In order to improve Compact Disc Players playability regarding playing Compact Discs with surface faults, like scratches and fingerprints etc, the attention has been put on fault tolerant control schemes. Almost every of those methods are based on fault detection. The standard approach is to use...... a pair of residuals generated by Compact Disc Player. However, these residuals depend on the performance of position servos in the Compact Disc Player. In other publications of the same authors a pair of decoupled residuals is derived. However, the computation of these alternative residuals has been...

  6. Optimization of the enzymatic extraction process of protein in tea residue by response surface methodology%响应面优化酶法提取茶渣蛋白的工艺研究

    Institute of Scientific and Technical Information of China (English)

    李圆圆; 王洪新; 殷飞; 兰海崇; 佘世凤

    2013-01-01

    To optimize the extraction parameters of protein in tea residue,the Central Composite Design was used to analyze the factors that affect the protein extraction rate.Alcalase was selected to extract the protein in tea residue. Based on single factor experiment results,enzyme hydrolysis temperature,reaction time,the ratio of liquid to solid, enzyme amount and enzyme hydrolysis pH were chosen as influencing factors and the protein yield was selected as the response value. The results showed that the optimal extraction conditions were: enzyme hydrolysis temperature 60℃,reaction time 1.5h,the ratio of liquid to solid 20:1,enzyme amount 2.5% and enzyme hydrolysis pH9.5.ln this optimal process condition,the yield of protein in tea residue was 56.95%.Compared with the previous report,extraction rate of protein in tea residue increased significantly with the application of response surface analysis methodology.%以茶渣为原料,选用碱性蛋白酶,利用中心组合实验设计对影响茶渣蛋白提取率的因素水平进行综合研究,优化茶渣蛋白的提取工艺参数根据单因素实验结果,以提取温度、提取时间、液固比、加酶量和pH为影响因子,茶渣蛋白提取率为响应值,确定最优提取条件为:提取温度60℃,提取时间1.5h,液固比20∶1,加酶量2.5%,pH9.5,此时茶渣蛋白提取率为56.95% 与之前报道相比,应用响应面分析法研究酶法提取茶渣蛋白,提取率有显著提高.

  7. Engineering out motion: a surface disulfide bond alters the mobility of tryptophan 22 in cytochrome b5 as probed by time-resolved fluorescence and 1H NMR experiments.

    Science.gov (United States)

    Storch, E M; Grinstead, J S; Campbell, A P; Daggett, V; Atkins, W M

    1999-04-20

    In the accompanying paper [Storch et al. (1999) Biochemistry 38, 5054-5064] equilibrium denaturation studies and molecular dynamics (MD) simulations were used to investigate localized dynamics on the surface of cytochrome b5 (cyt b5) that result in the formation of a cleft. In those studies, an S18C:R47C disulfide mutant was engineered to inhibit cleft mobility. Temperature- and urea-induced denaturation studies revealed significant differences in Trp 22 fluorescence between the wild-type and mutant proteins. On the basis of the results, it was proposed that wild type populates a conformational ensemble that is unavailable to the disulfide mutant and is mediated by cleft mobility. As a result, the solvent accessibility of Trp 22 is decreased in S18C:R47C, suggesting that the local environment of this residue is less mobile due to the constraining effects of the disulfide on cleft dynamics. To further probe the structural effects on the local environment of Trp 22 caused by inhibition of cleft formation, we report here the results of steady-state and time-resolved fluorescence quenching, differential phase/modulation fluorescence anisotropy, and 1H NMR studies. In Trp fluorescence experiments, the Stern-Volmer quenching constant increases in wild type versus the oxidized disulfide mutant with increasing temperature. At 50 degrees C, KSV is nearly 1.5-fold greater in wild type compared to the oxidized disulfide mutant. In the reduced disulfide mutant, KSV was the same as wild type. The bimolecular collisional quenching constant, kq, for acrylamide quenching of Trp 22 increases 2.7-fold for wild type and only 1.8-fold for S18C:R47C, upon increasing the temperature from 25 to 50 degrees C. The time-resolved anisotropy decay at 25 degrees C was fit to a double-exponential decay for both the wild type and S18C:R47C. Both proteins exhibited a minor contribution from a low-amplitude fast decay, consistent with local motion of Trp 22. This component was more prevalent in

  8. Modulation of innate and learned sexual behaviors by the TRP channel Painless expressed in the fruit fly brain: behavioral genetic analysis and its implications

    Directory of Open Access Journals (Sweden)

    Shoma eSato

    2014-12-01

    Full Text Available Transient receptor potential (TRP channels have attracted considerable attention because of their vital roles in primary sensory neurons, mediating responses to a wide variety of external environmental stimuli. However, much less is known about how TRP channels in the brain respond to intrinsic signals and are involved in neurophysiological processes that control complex behaviors. Painless (Pain is the Drosophila TRP channel that was initially identified as a molecular sensor responsible for detecting noxious thermal and mechanical stimuli. Here, we review recent behavioral genetic studies demonstrating that Pain expressed in the brain plays a critical role in both innate and learned aspects of sexual behaviors. Several members of the TRP channel superfamily play evolutionarily conserved roles in sensory neurons as well as in other peripheral tissues. It is thus expected that brain TRP channels in vertebrates and invertebrates would have some common physiological functions. Studies of Pain in the Drosophila brain using a unique combination of genetics and physiological techniques should provide valuable insights into the fundamental principles concerning TRP channels expressed in the vertebrate and invertebrate brains.

  9. Optimization of biogas residue mixed with livestock manure co-composting using response surface methodology%响应曲面法优化沼渣混合物料堆肥配比研究

    Institute of Scientific and Technical Information of China (English)

    宋彩红; 夏训峰; 席北斗; 魏自民; 李鸣晓; 何小松; 党秋玲

    2012-01-01

    According to principal components analysis, a comprehensive evaluation function was built with WSC/WSN, ?/^6. NH4+-N/NC>3~-N, GI, and the comprehensive evaluation value F used for the assessment of co-compost maturity was obtained on the basis of this evaluation function. In order to get the optimal proportion of biogas residue mixed with livestock manure co-composting, choosing F value as the response value and amount of biogas residue, pig manure, and chicken manure as the three factors, fifteen composting experiments were designed using Box-Behnken, and a second order polynomial equation was established. The results showed, the significant differences between the response value F and livestock manure amount was as follows: chicken manure > biogas residue > pig manure. Linear term of chicken manure amount and quadratic term of anaerobic digestion residue amount both reached extremely significant level (Z"<0.01). Interaction term of biogas residue amount ,chicken manure amount and quadratic term of chicken manure amount reached significant level (/*<0.05).The optimized proportion of co-compost was: the biogas residue amount 5.83kg, the pig manure amount 7.95kg, and the chicken manure amount 7.54kg. Therefore, it is reasonable that response surface method was used for optimization of material proportion of co-compost.%基于主成分分析,以WSC/WSN、E4/E6、NH4+-N/NO3--N、GI作为混合堆肥腐熟程度的判别指标体系,建立综合评价函数,确定综合评价值F,用于表征混合堆肥腐熟度.采用Box-Behnken设计,以F值为响应值,沼渣量、猪粪量、鸡粪量为3因素,设计了15组堆肥试验,建立二次多项数学模型,从腐熟度的角度确定沼渣、猪粪、鸡粪混合堆肥的最佳配比.结果表明:沼渣量、猪粪量、鸡粪量对响应值F影响的显著性顺序为:鸡粪量>沼渣量>猪粪量;鸡粪量一次项、沼渣量二次项达到极显著水平(P<0.01);沼渣量和鸡粪量的交互项、鸡粪

  10. Mutagenesis of bacteriophage IKe major coat protein transmembrane domain: role of an interfacial proline residue.

    Science.gov (United States)

    Williams, K A; Deber, C M

    1993-10-15

    The transmembrane (TM) domain of the 53-residue major coat protein of the M13-related bacteriophage IKe (residues 24-42: LISQTWPVVTTVVVAGVLI) has been subjected to randomized mutagenesis to probe the conformation and stability of the TM domain, as well as the effect of structurally-important residues such as proline. TM mutants were obtained by the Eckstein method of site-directed mutagenesis using the IKe genome as template so as to eliminate the need for subcloning. Over 40 single- and double-site viable mutants of bacteriophage IKe were isolated. Every residue in the TM segment, except the highly conserved Trp29, could be mutated to at least one other residue; polar and charged mutations occurred in the TM segment adjacent to the N-terminal domain (residues 24-28), while non-polar substitutions predominated in the C-terminal portion (residues 30-42). The Pro30 locus tolerated four mutations-Ala, Gly, Cys, and Ser- which represent the four side chains of least volume. Mutant coat proteins obtained directly from the phage in milligram quantities were studied by circular dichroism spectroscopy and SDS-PAGE gels. Wild type IKe coat protein solubilized in sodium deoxycholate micelles was found to occur as an alpha-helical, monomeric species which is stable at 95 degrees C, whereas the mutant Pro30-->Gly undergoes an irreversible conformational transition at ca. 90 degrees C to an aggregated beta-sheet structure. The result that Pro30 stabilizes the TM helix in the micellar membrane suggests a sterically-restricted location for the wild type Pro pyrrolidine side chain in the bulky Trp-Pro-Val triad, where it may be positioned to direct the initiation of the subsequent TM core domain helix.

  11. Structural and population-based evaluations of TBC1D1 p.Arg125Trp.

    Directory of Open Access Journals (Sweden)

    Tom G Richardson

    Full Text Available Obesity is now a leading cause of preventable death in the industrialised world. Understanding its genetic influences can enhance insight into molecular pathogenesis and potential therapeutic targets. A non-synonymous polymorphism (rs35859249, p.Arg125Trp in the N-terminal TBC1D1 phosphotyrosine-binding (PTB domain has shown a replicated association with familial obesity in women. We investigated these findings in the Avon Longitudinal Study of Parents and Children (ALSPAC, a large European birth cohort of mothers and offspring, and by generating a predicted model of the structure of this domain. Structural prediction involved the use of three separate algorithms; Robetta, HHpred/MODELLER and I-TASSER. We used the transmission disequilibrium test (TDT to investigate familial association in the ALSPAC study cohort (N = 2,292 mother-offspring pairs. Linear regression models were used to examine the association of genotype with mean measurements of adiposity (Body Mass Index (BMI, waist circumference and Dual-energy X-ray absorptiometry (DXA assessed fat mass, and logistic regression was used to examine the association with odds of obesity. Modelling showed that the R125W mutation occurs in a location of the TBC1D1 PTB domain that is predicted to have a function in a putative protein:protein interaction. We did not detect an association between R125W and BMI (mean per allele difference 0.27 kg/m(2 (95% Confidence Interval: 0.00, 0.53 P = 0.05 or obesity (odds ratio 1.01 (95% Confidence Interval: 0.77, 1.31, P = 0.96 in offspring after adjusting for multiple comparisons. Furthermore, there was no evidence to suggest that there was familial association between R125W and obesity (χ(2 = 0.06, P = 0.80. Our analysis suggests that R125W in TBC1D1 plays a role in the binding of an effector protein, but we find no evidence that the R125W variant is related to mean BMI or odds of obesity in a general population sample.

  12. Acrylonitrile quenching of trp phosphorescence in proteins: a probe of the internal flexibility of the globular fold.

    Science.gov (United States)

    Strambini, Giovanni B; Gonnelli, Margherita

    2010-08-04

    Quenching of Trp phosphorescence in proteins by diffusion of solutes of various molecular sizes unveils the frequency-amplitude of structural fluctuations. To cover the sizes gap between O(2) and acrylamide, we examined the potential of acrylonitrile to probe conformational flexibility of proteins. The distance dependence of the through-space acrylonitrile quenching rate was determined in a glass at 77 K, with the indole analog 2-(3-indoyl) ethyl phenyl ketone. Intensity and decay kinetics data were fitted to a rate, k(r) =k(0) exp[-(r -r(0))/r(e)], with an attenuation length r(e) = 0.03 nm and a contact rate k(0) = 3.6 x 10(10) s(-1). At ambient temperature, the bimolecular quenching rate constant (kq) was determined for a series of proteins, appositely selected to test the importance of factors such as the degree of Trp burial and structural rigidity. Relative to kq = 1.9 x 10(9) M(-1)s(-1) for free Trp in water, in proteins kq ranged from 6.5 x 10(6) M(-1)s(-1) for superficial sites to 1.3 x 10(2) M(-1)s(-1) for deep cores. The short-range nature of the interaction and the direct correlation between kq and structural flexibility attest that in the microsecond-second timescale of phosphorescence acrylonitrile readily penetrates even compact protein cores and exhibits significant sensitivity to variations in dynamical structure of the globular fold.

  13. Kinetics of formation for lanthanide (III) complexes of DTPA-(Me-Trp)2 used as imaging agent.

    Science.gov (United States)

    Tiwari, Anjani K; Sinha, Deepa; Datta, Anupama; Kakkar, Dipti; Mishra, Anil K

    2011-05-01

    Diethlenetriamine-N,N,N'N''N''-pentaacetic acid (DTPA)-bis (amide) analogs have been synthesized and evaluated as a potential biomedical imaging agents. Imaging and biodistribution studies were performed in mice that showed a significant accumulation of DTPA analogs in brain. The stability and protonation constants of the complexes formed between the ligand [DTPA-(Me-Trp)(2)] and Gd(3+), Eu(3+), and Cu(2+) have been determined by pH potentiometry (Gd(3+), Eu(3+)) and spectrophotometry (Cu(2+)) at 25 °C and at constant ionic strength maintained by 0.10 M KCl. The kinetic inertness of Gd [DTPA-(Me-Trp)(2)] was characterized by the rates of exchange reactions with Zn(2+) and Eu(3+). In the Eu(3+) exchange, a second-order [H(+)] dependence was found for the pseudo-first-order rate constant [k(0) = (4.5 ± 1.2) × 10(-6)/s; k(1) = 0.58 ± 0.1 /M/s, k(2) = (6.6 ± 0.2) × 10(4) /M(2)/s, k(3) = (4.8 ± 0.8) × 10(-4) /M/s]. In the Eu(3+) exchange, at pH DTPA-(Me-Trp)(2)] is more inert than GdDTPA(2-), the most commonly used MRI contrast agent (t(1/2) = 127 h). High kinetic stability is an important requirement for the Gd complexes used as contrast enhancement agents in magnetic resonance imaging. © 2011 John Wiley & Sons A/S.

  14. Investigation of KIF6 Trp719Arg in a case-control study of myocardial infarction: a Costa Rican population.

    Directory of Open Access Journals (Sweden)

    Lance A Bare

    Full Text Available BACKGROUND AND METHODOLOGY: The 719Arg allele of KIF6 (rs20455 was associated with coronary events in Caucasian participants of five prospective studies. We investigated whether this KIF6 variant was associated with non-fatal myocardial infarction (MI in a case-control study of an admixed population from the Central Valley of Costa Rica. Genotypes of the KIF6 variant were determined for 4,134 men and women. Cases (1,987 had survived a first MI; controls (2,147 had no history of MI and were matched to cases by age, sex, and area of residence. We tested the association between the KIF6 719Arg allele and non-fatal MI by conditional logistic regression and adjusted for admixture of founder populations. PRINCIPAL FINDINGS: Compared with the reference Trp/Trp homozygotes, KIF6 719Arg carriers were not at significantly higher risk for non-fatal MI in this study after adjustment for traditional risk factors or admixture (OR= 1.12; 95%CI, 0.98-1.28. Heterozygotes of the KIF6 Trp719Arg variant were at increased risk of non-fatal MI: the adjusted odds ratio was 1.16 (95% confidence interval, 1.01-1.34, but this association would not be significant after a multiple testing correction. CONCLUSIONS/SIGNIFICANCE: We found that carriers of the KIF6 719Arg allele were not at increased risk of non-fatal MI in a case-control study of Costa Ricans living in the Central Valley of Costa Rica.

  15. Alphavirus replicon particles expressing TRP-2 provide potent therapeutic effect on melanoma through activation of humoral and cellular immunity.

    Directory of Open Access Journals (Sweden)

    Francesca Avogadri

    Full Text Available BACKGROUND: Malignant melanoma is the deadliest form of skin cancer and is refractory to conventional chemotherapy and radiotherapy. Therefore alternative approaches to treat this disease, such as immunotherapy, are needed. Melanoma vaccine design has mainly focused on targeting CD8+ T cells. Activation of effector CD8+ T cells has been achieved in patients, but provided limited clinical benefit, due to immune-escape mechanisms established by advanced tumors. We have previously shown that alphavirus-based virus-like replicon particles (VRP simultaneously activate strong cellular and humoral immunity against the weakly immunogenic melanoma differentiation antigen (MDA tyrosinase. Here we further investigate the antitumor effect and the immune mechanisms of VRP encoding different MDAs. METHODOLOGY/PRINCIPAL FINDINGS: VRP encoding different MDAs were screened for their ability to prevent the growth of the B16 mouse transplantable melanoma. The immunologic mechanisms of efficacy were investigated for the most effective vaccine identified, focusing on CD8+ T cells and humoral responses. To this end, ex vivo immune assays and transgenic mice lacking specific immune effector functions were used. The studies identified a potent therapeutic VRP vaccine, encoding tyrosinase related protein 2 (TRP-2, which provided a durable anti-tumor effect. The efficacy of VRP-TRP2 relies on a novel immune mechanism of action requiring the activation of both IgG and CD8+ T cell effector responses, and depends on signaling through activating Fcγ receptors. CONCLUSIONS/SIGNIFICANCE: This study identifies a VRP-based vaccine able to elicit humoral immunity against TRP-2, which plays a role in melanoma immunotherapy and synergizes with tumor-specific CD8+ T cell responses. These findings will aid in the rational design of future immunotherapy clinical trials.

  16. Alphavirus replicon particles expressing TRP-2 provide potent therapeutic effect on melanoma through activation of humoral and cellular immunity.

    Science.gov (United States)

    Avogadri, Francesca; Merghoub, Taha; Maughan, Maureen F; Hirschhorn-Cymerman, Daniel; Morris, John; Ritter, Erika; Olmsted, Robert; Houghton, Alan N; Wolchok, Jedd D

    2010-09-10

    Malignant melanoma is the deadliest form of skin cancer and is refractory to conventional chemotherapy and radiotherapy. Therefore alternative approaches to treat this disease, such as immunotherapy, are needed. Melanoma vaccine design has mainly focused on targeting CD8+ T cells. Activation of effector CD8+ T cells has been achieved in patients, but provided limited clinical benefit, due to immune-escape mechanisms established by advanced tumors. We have previously shown that alphavirus-based virus-like replicon particles (VRP) simultaneously activate strong cellular and humoral immunity against the weakly immunogenic melanoma differentiation antigen (MDA) tyrosinase. Here we further investigate the antitumor effect and the immune mechanisms of VRP encoding different MDAs. VRP encoding different MDAs were screened for their ability to prevent the growth of the B16 mouse transplantable melanoma. The immunologic mechanisms of efficacy were investigated for the most effective vaccine identified, focusing on CD8+ T cells and humoral responses. To this end, ex vivo immune assays and transgenic mice lacking specific immune effector functions were used. The studies identified a potent therapeutic VRP vaccine, encoding tyrosinase related protein 2 (TRP-2), which provided a durable anti-tumor effect. The efficacy of VRP-TRP2 relies on a novel immune mechanism of action requiring the activation of both IgG and CD8+ T cell effector responses, and depends on signaling through activating Fcγ receptors. This study identifies a VRP-based vaccine able to elicit humoral immunity against TRP-2, which plays a role in melanoma immunotherapy and synergizes with tumor-specific CD8+ T cell responses. These findings will aid in the rational design of future immunotherapy clinical trials.

  17. Expression of the Trp2 allele of COL9A2 is associated with alterations in the mechanical properties of human intervertebral discs.

    Science.gov (United States)

    Aladin, Darwesh M K; Cheung, Kenneth M C; Chan, Danny; Yee, Anita F Y; Jim, Jeffrey J T; Luk, Keith D K; Lu, William W

    2007-12-01

    Biomechanical study into the association between genetic polymorphism in COL9A2 and mechanical properties of human nucleus pulposus. To examine whether there is an association between genetic polymorphism in a structural gene, and alterations in the mechanical properties of the intervertebral discs that may predispose to disc degeneration. Genetic studies have demonstrated that a polymorphism (Trp2 allele) in COL9A2 coding for alpha2 chain of collagen IX predisposes the individual to disc degeneration. The mechanism of this predisposition is not known. Blood and whole disc samples were retrieved from adolescents and young adults during scoliosis surgery, degenerated discs were retrieved from patients with back pain during anterior spinal fusion. Anulus fibrosus and nucleus pulposus from a set of the scoliosis discs were used to perform immunohistochemistry to demonstrate the presence of collagen IX in the scoliosis discs. For the remaining samples, DNA was extracted from blood to determine the Trp2 status by sequencing. Nondegenerated (Trp2-), nondegenerated (Trp2+), and degenerated (Trp2-) nucleus pulposus samples were tested in confined compression. Swelling pressure and compressive modulus were measured and compared between groups. Positive staining of collagen IX was detected in both anulus fibrosus and nucleus pulposus sections confirming its presence in the scoliosis discs. The mean swelling pressure and compressive modulus values of 6 nondegenerated (Trp2+) samples (swelling pressure = 0.0019 MPa, compressive modulus = 0.97 MPa) were significantly lower (P pressure = 0.015 MPa; compressive modulus = 1.89 MPa). This is the first study to demonstrate an association between the Trp2 allele and disc mechanics, thus relating genetic variations and debilitating mechanical alterations that may ultimately result in intervertebral disc degeneration.

  18. Análisis preliminar de la estructura primaria y secundaria del ARNtTrp en tortugas marinas

    OpenAIRE

    Harvey Infante-Rojas; Javier Hernández-Fernández

    2015-01-01

    Actualmente existen siete especies de tortugas marinas, todas amenazadas o en riesgo inminente de extinción. Los estudios con ADN mitocondrial han permitido hacer acercamientos sobre filogenia, evolución, rutas migratorias y centros de dispersión, además para la identificación de polimorfismos y haplotipos, siendo base para planes de manejo y conservación. El presente estudio representa la primera descripción comparada de la estructura primaria y secundaria del ARNtTrp mitocondrial en tortuga...

  19. Effects of rhGM-CSF hydrogel on the healing of residual burn wound surface%rhGM-CSF凝胶对烧伤后残余创面愈合的影响

    Institute of Scientific and Technical Information of China (English)

    邱学文; 王甲汉; 杨磊; 任加良

    2011-01-01

    Objective To investigate the effects of recombinam human granulocyte macrophage colony-stimulating factor (rhGMCSF) hydrogel on the healing of residual burn wound surface. Methods One hundred and thirty-eight residual bum wound surfaces were assigned into experimental group and control group (69 each), and randomized, double-blind, placebo-controlled and self-controlled clinical trials were conducted. The wound surfaces in experimental group were treated with rhGM-CSF hydrogel, and in control group with placebo, for 21 days. The healing time, therapeutic effects, and the healing rate of the wound surfaces at day 7 and day 14 after treatment were observed. The tissue samples were harvested from the border of the residual wounds from 6 patients on the 7th and 14th day after treatment, and the number of blood capillaries and fibroblasts was counted. Results Nine out of sixty-nine patients with residual bum wounds were excluded from the study, the data of only 60 patients were analyzed. The mean healing time of wounds was 12 days (95%CI 11-13 days) in experimental group, and it was significantly shorter than that in control group (18 days, 95%CI 17-19 days, P〈0.001);meanwhile the therapeutic effects of experimental group was significantly better than that of control group (P<0.001). The rate of healing in experimental group was 62%±13% and 95%±10%, respectively, on the 7th and 14th day after treatment, and they were significantly higher than that in control group (46% ±11% and 83% ± 12%, respectively, P<0. 001). At the 7th and 14th day after treatment,histopathological examination showed that the counts of capillaries (10.3±0.6/HP and 14.5 ±0.9/HP, respectively) and fibroblasts (143.1± 10.3/HP and 137.8 ± 6.9/HP, respectively) in experimental group were significantly higher than those in control group (capillaries: 7.2±0.7 and 10.4±0.8, P<0.01; fibroblasts.. 110.2±11.7 and 126.4±7.7, P<0.01). Conclusions Topical use of rh

  20. Interaction of disinfectant residues on cleanroom substrates.

    Science.gov (United States)

    Kaiser, H; Klein, D; Kopis, E; Leblanc, D; McDonnell, G; Tirey, J F

    1999-01-01

    This study will determine the levels of disinfectant residues on stainless steel surfaces after simulated manual cleaning activities. Additionally, this study will determine if chemical interactions between different chemical agents, representative of commonly used cleanroom disinfectant technologies, subsequently applied to the same surfaces exist, and to what degree these interactions impact sporicidal performance of an oxidizing biocide against Bacillus subtilis.

  1. Novel intracellular GH10 xylanase from Cohnella laeviribosi HY-21: biocatalytic properties and alterations of substrate specificities by site-directed mutagenesis of Trp residues.

    Science.gov (United States)

    Kim, Do Young; Han, Mi Kyoung; Oh, Hyun-Woo; Bae, Kyung Sook; Jeong, Tae-Sook; Kim, Sung Uk; Shin, Dong-Ha; Kim, In-Ho; Rhee, Young Ha; Son, Kwang-Hee; Park, Ho-Yong

    2010-11-01

    The novel intracellular GH10 xylanase (iXylC) gene (1023-bp) of Cohnella laeviribosi HY-21 encoded a protein consisting of 340 amino acids with a deduced molecular mass of 39,330Da and a calculated pI of 5.81. The primary structure of iXylC was 70% identical to that of Geobacillus sp. GH10 enzyme (GenBank accession number: EDV78425). Xylanolytic activity of the His-tagged iXylC overproduced in Escherichiacoli BL21 was stimulated by 2.2-fold in the presence of 0.5% non-ionic detergents. iXylC produced a mixture of xylooligosaccharides (xylobiose to xylooctaose) from xylotriose and xylotetraose used as the hydrolytic substrate. In addition, it exhibited considerable cleavage activities for p-nitrophenylxylopyranoside (PNP-xylopyranoside) and PNP-cellobioside, indicating that iXylC is a unique GH10 enzyme. The hydrolytic activity (57.8IUmL(-1)) of iXylC toward PNP-xylopyranoside increased to 8.3-fold by W217A and W315A mutations, while mutations of W133A, W295A, and W303A abolished the hydrolytic activity of the enzyme.

  2. Advances in research of the effects of menthol on TRP channels%薄荷醇影响TRP通道的研究进展

    Institute of Scientific and Technical Information of China (English)

    黄钊; 王晖

    2011-01-01

    In recent years it has been discovered that there is a close connection between menthol and the transient receptor potential (TRP) channels, including TRPV1, TRPV3, TRPM8 and TRPA1. This review attempts to summarize the advances in research of the effects of menthol on the TRP channels regulating the intracellular calcium ion.%近年来,国内外学者在薄荷醇影响钙离子通道的研究中发现,其与瞬时感受器电位(TRP)通道有着密切的关系,其中包括TRPV1、TRPV3、TRPM8和TRPA1.本文对近年来薄荷醇参与细胞内Ca2+调节的TRP通道的相关研究进行综述.

  3. TrpM, a Small Protein Modulating Tryptophan Biosynthesis and Morpho-Physiological Differentiation in Streptomyces coelicolor A3(2)

    Science.gov (United States)

    Palazzotto, Emilia; Gallo, Giuseppe; Renzone, Giovanni; Giardina, Anna; Sutera, Alberto; Silva, Joohee; Vocat, Celinè; Botta, Luigi; Scaloni, Andrea; Puglia, Anna Maria

    2016-01-01

    In the model actinomycete Streptomyces coelicolor A3(2), small open reading frames encoding proteins with unknown functions were identified in several amino acid biosynthetic gene operons, such as SCO2038 (trpX) in the tryptophan trpCXBA locus. In this study, the role of the corresponding protein in tryptophan biosynthesis was investigated by combining phenotypic and molecular analyses. The 2038KO mutant strain was characterized by delayed growth, smaller aerial hyphae and reduced production of spores and actinorhodin antibiotic, with respect to the WT strain. The capability of this mutant to grow on minimal medium was rescued by tryptophan and tryptophan precursor (serine and/or indole) supplementation on minimal medium and by gene complementation, revealing the essential role of this protein, here named TrpM, as modulator of tryptophan biosynthesis. His-tag pull-down and bacterial adenylate cyclase-based two hybrid assays revealed TrpM interaction with a putative leucyl-aminopeptidase (PepA), highly conserved component among various Streptomyces spp. In silico analyses showed that PepA is involved in the metabolism of serine, glycine and cysteine through a network including GlyA, CysK and CysM enzymes. Proteomic experiments suggested a TrpM-dependent regulation of metabolic pathways and cellular processes that includes enzymes such as GlyA, which is required for the biosynthesis of tryptophan precursors and key proteins participating in the morpho-physiological differentiation program. Altogether, these findings reveal that TrpM controls tryptophan biosynthesis at the level of direct precursor availability and, therefore, it is able to exert a crucial effect on the morpho-physiological differentiation program in S. coelicolor A3(2). PMID:27669158

  4. Residual stress profiling of an aluminum alloy by laser ultrasonics

    Institute of Scientific and Technical Information of China (English)

    PAN Yondong; QIAN Menglu; XU Weijiang; M. OURAK

    2004-01-01

    A residual-stress profile along the thickness of an aluminum alloy sheet is determined by laser-ultrasonic technique. Surface acoustic waves are generated by a Nd:YAG pulse laser and detected by a Heterodyne interferometer on a lateral free surface of the sheet. The distribution of residual stress is determined by measuring the relative variation of the wavevelocities at different location of the sample along its thickness. This technique is validated by three different residual stress profiles obtained experimentally.

  5. 一种解决小型终端手机天线的人头人手OTA指标(TRP&TIS)的方法

    Institute of Scientific and Technical Information of China (English)

    吴诚君

    2011-01-01

    1.更改天线馈地点位置 2.天线下方PCB板上增加地 3.利用最小面积调试天线 本专利申请的名称: 一种解决小型终端手机天线的人头人手OTA指标(TRP&TIS)的方法 要解决的技术问题是什么: 本发明:1.解决了手机天线的人头人手指标(TRP&TIS),满足TRP和TIS指标。

  6. Structural insights into cellulolytic and chitinolytic enzymes revealing crucial residues of insect β-N-acetyl-D-hexosaminidase.

    Directory of Open Access Journals (Sweden)

    Tian Liu

    Full Text Available The chemical similarity of cellulose and chitin supports the idea that their corresponding hydrolytic enzymes would bind β-1,4-linked glucose residues in a similar manner. A structural and mutational analysis was performed for the plant cellulolytic enzyme BGlu1 from Oryza sativa and the insect chitinolytic enzyme OfHex1 from Ostrinia furnacalis. Although BGlu1 shows little amino-acid sequence or topological similarity with OfHex1, three residues (Trp(490, Glu(328, Val(327 in OfHex1, and Trp(358, Tyr(131 and Ile(179 in BGlu1 were identified as being conserved in the +1 sugar binding site. OfHex1 Glu(328 together with Trp(490 was confirmed to be necessary for substrate binding. The mutant E328A exhibited a 8-fold increment in K(m for (GlcNAc(2 and a 42-fold increment in K(i for TMG-chitotriomycin. A crystal structure of E328A in complex with TMG-chitotriomycin was resolved at 2.5 Å, revealing the obvious conformational changes of the catalytic residues (Glu(368 and Asp(367 and the absence of the hydrogen bond between E328A and the C3-OH of the +1 sugar. V327G exhibited the same activity as the wild-type, but acquired the ability to efficiently hydrolyse β-1,2-linked GlcNAc in contrast to the wild-type. Thus, Glu(328 and Val(327 were identified as important for substrate-binding and as glycosidic-bond determinants. A structure-based sequence alignment confirmed the spatial conservation of these three residues in most plant cellulolytic, insect and bacterial chitinolytic enzymes.

  7. Computational analysis of residue contributions to coiled-coil topology.

    Science.gov (United States)

    Ramos, Jorge; Lazaridis, Themis

    2011-11-01

    A variety of features are thought to contribute to the oligomeric and topological specificity of coiled coils. In previous work, we examined the determinants of oligomeric state. Here, we examine the energetic basis for the tendency of six coiled-coil peptides to align their α-helices in antiparallel orientation using molecular dynamics simulations with implicit solvation (EEF1.1). We also examine the effect of mutations known to disrupt the topology of these peptides. In agreement with experiment, ARG or LYS at a or d positions were found to stabilize the antiparallel configuration. The modeling suggests that this is not due to a-a' or d-d' repulsions but due to interactions with e' and g' residues. TRP at core positions also favors the antiparallel configuration. Residues that disfavor parallel dimers, such as ILE at d, are better tolerated in, and thus favor the antiparallel configuration. Salt bridge networks were found to be more stabilizing in the antiparallel configuration for geometric reasons: antiparallel helices point amino acid side chains in opposite directions. However, the structure with the largest number of salt bridges was not always the most stable, due to desolvation and configurational entropy contributions. In tetramers, the extent of stabilization of the antiparallel topology by core residues is influenced by the e' residue on a neighboring helix. Residues at b and c positions in some cases also contribute to stabilization of antiparallel tetramers. This work provides useful rules toward the goal of designing coiled coils with a well-defined and predictable three-dimensional structure.

  8. Trp RNA-binding attenuation protein: modifying symmetry and stability of a circular oligomer.

    Directory of Open Access Journals (Sweden)

    Oliver W Bayfield

    Full Text Available BACKGROUND: Subunit number is amongst the most important structural parameters that determine size, symmetry and geometry of a circular protein oligomer. The L-tryptophan biosynthesis regulator, TRAP, present in several Bacilli, is a good model system for investigating determinants of the oligomeric state. A short segment of C-terminal residues defines whether TRAP forms an 11-mer or 12-mer assembly. To understand which oligomeric state is more stable, we examine the stability of several wild type and mutant TRAP proteins. METHODOLOGY/PRINCIPAL FINDINGS: Among the wild type B. stearothermophilus, B. halodurans and B. subtilis TRAP, we find that the former is the most stable whilst the latter is the least. Thermal stability of all TRAP is shown to increase with L-tryptophan concentration. We also find that mutant TRAP molecules that are truncated at the C-terminus - and hence induced to form 12-mers, distinct from their 11-mer wild type counterparts--have increased melting temperatures. We show that the same effect can be achieved by a point mutation S72N at a subunit interface, which leads to exclusion of C-terminal residues from the interface. Our findings are supported by dye-based scanning fluorimetry, CD spectroscopy, and by crystal structure and mass spectrometry analysis of the B. subtilis S72N TRAP. CONCLUSIONS/SIGNIFICANCE: We conclude that the oligomeric state of a circular protein can be changed by introducing a point mutation at a subunit interface. Exclusion (or deletion of the C-terminus from the subunit interface has a major impact on properties of TRAP oligomers, making them more stable, and we argue that the cause of these changes is the altered oligomeric state. The more stable TRAP oligomers could be used in potential applications of TRAP in bionanotechnology.

  9. Phytochemicals from Ruta graveolens Activate TAS2R Bitter Taste Receptors and TRP Channels Involved in Gustation and Nociception

    Directory of Open Access Journals (Sweden)

    Giuseppe Mancuso

    2015-10-01

    Full Text Available Ruta graveolens (rue is a spontaneous plant in the Mediterranean area with a strong aroma and a very intense bitter taste, used in gastronomy and in folk medicine. From the leaves, stems and fruits of rue, we isolated rutin, rutamarin, three furanocoumarins, two quinolinic alkaloids, a dicoumarin and two long chain ketones. Bitter taste and chemesthetic properties have been evaluated by in vitro assays with twenty receptors of the TAS2R family and four TRP ion channels involved in gustation and nociception. Among the alkaloids, skimmianine was active as a specific agonist of T2R14, whereas kokusaginin did not activate any of the tested receptors. The furanocoumarins activates TAS2R10, 14, and 49 with different degrees of selectivity, as well as the TRPA1 somatosensory ion channel. Rutamarin is an agonist of TRPM5 and TRPV1 and a strong antagonist of TRPM8 ion channels.

  10. Endo-lysosomal TRP mucolipin-1 channels trigger global ER Ca2+ release and Ca2+ influx

    Science.gov (United States)

    Kilpatrick, Bethan S.; Yates, Elizabeth; Grimm, Christian; Schapira, Anthony H.

    2016-01-01

    ABSTRACT Transient receptor potential (TRP) mucolipins (TRPMLs), encoded by the MCOLN genes, are patho-physiologically relevant endo-lysosomal ion channels crucial for membrane trafficking. Several lines of evidence suggest that TRPMLs mediate localised Ca2+ release but their role in Ca2+ signalling is not clear. Here, we show that activation of endogenous and recombinant TRPMLs with synthetic agonists evoked global Ca2+ signals in human cells. These signals were blocked by a dominant-negative TRPML1 construct and a TRPML antagonist. We further show that, despite a predominant lysosomal localisation, TRPML1 supports both Ca2+ release and Ca2+ entry. Ca2+ release required lysosomal and ER Ca2+ stores suggesting that TRPMLs, like other endo-lysosomal Ca2+ channels, are capable of ‘chatter’ with ER Ca2+ channels. Our data identify new modalities for TRPML1 action. PMID:27577094

  11. Phytochemicals from Ruta graveolens Activate TAS2R Bitter Taste Receptors and TRP Channels Involved in Gustation and Nociception.

    Science.gov (United States)

    Mancuso, Giuseppe; Borgonovo, Gigliola; Scaglioni, Leonardo; Bassoli, Angela

    2015-10-16

    Ruta graveolens (rue) is a spontaneous plant in the Mediterranean area with a strong aroma and a very intense bitter taste, used in gastronomy and in folk medicine. From the leaves, stems and fruits of rue, we isolated rutin, rutamarin, three furanocoumarins, two quinolinic alkaloids, a dicoumarin and two long chain ketones. Bitter taste and chemesthetic properties have been evaluated by in vitro assays with twenty receptors of the TAS2R family and four TRP ion channels involved in gustation and nociception. Among the alkaloids, skimmianine was active as a specific agonist of T2R14, whereas kokusaginin did not activate any of the tested receptors. The furanocoumarins activates TAS2R10, 14, and 49 with different degrees of selectivity, as well as the TRPA1 somatosensory ion channel. Rutamarin is an agonist of TRPM5 and TRPV1 and a strong antagonist of TRPM8 ion channels.

  12. Optimized Combination of Residue Hydrodesulfurization and Residue Fluid Catalytic Cracking

    Institute of Scientific and Technical Information of China (English)

    Chen Junwu

    2003-01-01

    @@1 Introduction Combination of residue hydrodesulfurization (HDS) and resi-due fluid catalytic cracking (RFCC) is a unique technologyfor processing high-sulfur residue. This paper discusses theoptimized combination of these two processes.

  13. Design and surface immobilization of short anti-biofilm peptides.

    Science.gov (United States)

    Mishra, Biswajit; Lushnikova, Tamara; Golla, Radha M; Wang, Xiuqing; Wang, Guangshun

    2017-02-01

    Short antimicrobial peptides are essential to keep us healthy and their lasting potency can inspire the design of new types of antibiotics. This study reports the design of a family of eight-residue tryptophan-rich peptides (TetraF2W) obtained by converting the four phenylalanines in temporin-SHf to tryptophans. The temporin-SHf template was identified from the antimicrobial peptide database (http://aps.unmc.edu/AP). Remarkably, the double arginine variant (TetraF2W-RR) was more effective in killing methicillin-resistant Staphylococcus aureus (MRSA) USA300, but less cytotoxic to human skin HaCat and kidney HEK293 cells, than the lysine-containing dibasic combinations (KR, RK and KK). Killing kinetics and fluorescence spectroscopy suggest membrane targeting of TetraF2W-RR, making it more difficult for bacteria to develop resistance. Because established biofilms on medical devices are difficult to remove, we chose to covalently immobilize TetraF2W-RR onto the polyethylene terephthalate (PET) surface to prevent biofilm formation. The successful surface coating of the peptide is supported by FT-IR and XPS spectroscopies, chemical quantification, and antibacterial assays. This peptide-coated surface indeed prevented S. aureus biofilm formation with no cytotoxicity to human cells. In conclusion, TetraF2W-RR is a short Trp-rich peptide with demonstrated antimicrobial and anti-biofilm potency against MRSA in both the free and immobilized forms. Because these short peptides can be synthesized cost effectively, they may be developed into new antimicrobial agents or used as surface coating compounds.

  14. Axial residual stresses in boron fibers

    Science.gov (United States)

    Behrendt, D. R.

    1978-01-01

    A method of measuring axial residual stresses in boron fibers is presented. With this method, the axial residual stress distribution as a function of radius is determined from the fiber surface to the core including the average residual stress in the core. Such measurements on boron on tungsten (B/W) fibers show that the residual stresses for 102, 142, 203, and 366 micron diam fibers are similar, being compressive at the surface and changing monotonically to a region of tensile stress within the boron. At approximately 25% of the original radius, the stress reaches a maximum tensile stress of about 860 MN sq m and then decreases to a compressive stress near the tungsten boride core. Data are presented for 203-micron diam B/W fibers that show annealing above 900 C reduces the residual stresses. A comparison between 102-micron diam B/W and boron on carbon (B/C) show that the residual stresses are similar in the outer regions of the fibers, but that large differences near and in the core are observed. Fracture of boron fibers is discussed.

  15. TENORM: Coal Combustion Residuals

    Science.gov (United States)

    Burning coal in boilers to create steam for power generation and industrial applications produces a number of combustion residuals. Naturally radioactive materials that were in the coal mostly end up in fly ash, bottom ash and boiler slag.

  16. Residual stress distribution in rabbit limb bones.

    Science.gov (United States)

    Yamada, Satoshi; Tadano, Shigeru; Fujisaki, Kazuhiro

    2011-04-29

    The presence of the residual stresses in bone tissue has been noted and the authors have reported that there are residual stresses in bone tissue. The aim of our study is to measure the residual stress distribution in the cortical bone of the extremities of vertebrates and to describe the relationships with the osteon population density. The study used the rabbit limb bones (femur, tibia/fibula, humerus, and radius/ulna) and measured the residual stresses in the bone axial direction at anterior and posterior positions on the cortical surface. The osteons at the sections at the measurement positions were observed by microscopy. As a result, the average stresses at the hindlimb bones and the forelimb bones were 210 and 149 MPa, respectively. In the femur, humerus, and radius/ulna, the residual stresses at the anterior position were larger than those at the posterior position, while in the tibia, the stress at the posterior position was larger than that at the anterior position. Further, in the femur and humerus, the osteon population densities in the anterior positions were larger than those in the posterior positions. In the tibia, the osteon population density in the posterior position was larger than that in the anterior position. Therefore, tensile residual stresses were observed at every measurement position in the rabbit limb bones and the value of residual stress correlated with the osteon population density (r=0.55, P<0.01).

  17. Associations between Dietary Patterns, ADRβ2 Gln27Glu and ADRβ3 Trp64Arg with Regard to Serum Triglyceride Levels: J-MICC Study

    Directory of Open Access Journals (Sweden)

    Hinako Nanri

    2016-09-01

    Full Text Available Interactions between dietary patterns and 2 β-adrenergic receptor (ADRβ gene polymorphisms (ADRβ2 Gln27Glu and ADRβ3 Trp64Arg were examined with regard to the effects on serum triglyceride levels. The cross-sectional study comprised 1720 men and women (aged 35–69 years enrolled in the Japan Multi-Institutional Collaborative Cohort (J-MICC Study. Genotyping was conducted using a multiplex polymerase chain reaction-based invader assay. We used 46 items from a validated short food frequency questionnaire and examined major dietary patterns by factor analysis. We identified four dietary patterns: healthy, Western, seafood and bread patterns. There was no significant association between any dietary pattern and serum triglyceride levels. After a separate genotype-based analysis, significant interactions between ADRβ3 Trp64Arg genotype and the bread pattern (p for interaction = 0.01 were associated with serum triglyceride levels; specifically, after adjusting for confounding factors, Arg allele carriers with the bread pattern had lower serum triglycerides (p for trend = 0.01. However, the Trp/Trp homozygous subjects with the bread pattern showed no association with serum triglycerides (p for trend = 0.55. Interactions between other dietary patterns and ADRβ polymorphisms were not significant for serum triglyceride levels. Our findings suggest that ADRβ3 polymorphism modifies the effects of the bread pattern on triglyceride levels.

  18. The trpE gene negatively regulates differentiation of heterocysts at the level of induction in Anabaena sp. strain PCC 7120.

    Science.gov (United States)

    Videau, Patrick; Cozy, Loralyn M; Young, Jasmine E; Ushijima, Blake; Oshiro, Reid T; Rivers, Orion S; Burger, Andrew H; Callahan, Sean M

    2015-01-01

    Levels of 2-oxoglutarate (2-OG) reflect nitrogen status in many bacteria. In heterocystous cyanobacteria, a spike in the 2-OG level occurs shortly after the removal of combined nitrogen from cultures and is an integral part of the induction of heterocyst differentiation. In this work, deletion of one of the two annotated trpE genes in Anabaena sp. strain PCC 7120 resulted in a spike in the 2-OG level and subsequent differentiation of a wild-type pattern of heterocysts when filaments of the mutant were transferred from growth on ammonia to growth on nitrate. In contrast, 2-OG levels were unaffected in the wild type, which did not differentiate under the same conditions. An inverted-repeat sequence located upstream of trpE bound a central regulator of differentiation, HetR, in vitro and was necessary for HetR-dependent transcription of a reporter fusion and complementation of the mutant phenotype in vivo. Functional complementation of the mutant phenotype with the addition of tryptophan suggested that levels of tryptophan, rather than the demonstrated anthranilate synthase activity of TrpE, mediated the developmental response of the wild type to nitrate. A model is presented for the observed increase in 2-OG in the trpE mutant.

  19. Isolation of the neuropeptide less than Glu-Trp-Leu-Lys-Gly-Arg-Phe-NH2 (Pol-RFamide II) from the hydromedusa Polyorchis penicillatus

    DEFF Research Database (Denmark)

    Grimmelikhuijzen, C J; Rinehart, K L; Spencer, A N

    1992-01-01

    Using a radioimmunoassay for the sequence Arg-Phe-NH2 (RFamide), we have isolated the peptide less than Glu-Trp-Leu-Lys-Gly-Arg-Phe-NH2 (Pol-RFamide II) from acetic acid extracts of the hydromedusa Polyorchis penicillatus. This peptide is a neuropeptide and constitutes a peptide family together w...

  20. Modelling pesticides residues

    OpenAIRE

    2004-01-01

    This work is a contribution to the development of a specific method to assess the presence of residues in agricultural commodities. The following objectives are formulated: to identify and describe main processes in environment — plant exchanges, to build of a model to assess the residue concentration at harvest in agricultural commodities, to understand the functioning of the modelled system, to characterise pesticides used in field crops and identify optimisation potentials in phytosanitary...

  1. Aplicação superficial de calcário e diferentes resíduos em soja cultivada no sistema plantio direto Surface application of limestone and different residues on soybean grown in no-till system

    Directory of Open Access Journals (Sweden)

    Juliano Corulli Corrêa

    2009-01-01

    Full Text Available A prática da correção da acidez do solo pela aplicação superficial de corretivos sobre a palha no sistema plantio direto se restringe ao calcário, não havendo maiores estudos em relação à escória de aciaria, lama cal e lodo de esgoto centrifugado. O objetivo do trabalho foi avaliar os índices de acidez do solo e a produtividade da soja em função da aplicação superficial de lodo de esgoto centrifugado, lama cal, escória de aciaria e calcário dolomítico. O trabalho foi desenvolvido em Latossolo Vermelho distrófico, durante os anos agrícolas de 2002 a 2005, sob sistema plantio direto. Os tratamentos constituíram da aplicação superficial de escória de aciaria (E, lama cal (Lcal, lodo de esgoto centrifugado (LC, calcário dolomítico e sem aplicação de corretivo, em delineamento em blocos ao acaso, com quatro repetições. A aplicação superficial de escória de aciaria, lama cal e lodo de esgoto centrifugado permite a correção da acidez do solo, o deslocamento do Ca2+, o aumento da saturação por bases e redução do Al3+ até 40 cm, e para o calcário, até 20 cm, fatores que condicionaram o aumento da produtividade da soja para os tratamentos LC, E e Lcal em 2003/2004 e 2004/2005 e para LC e E em 2002/2003 no sistema plantio direto.The practice of correcting soil acidity by surface application of pH-correcting materials on crop residues in the no-till system is restricted to limestone. No further studies are available on the use of steel slag, lime mud, and centrifuged sewage sludge. The objective of this study was to evaluate soil acidity and yield of soybean as a function of surface application of centrifuged sewage sludge, lime mud, steel slag, and dolomitic limestone. The study was conducted on a dystrophic Clayey Rhodic Hapludox soil, during the 2002_2005 cropping seasons, under notill system. Treatments consisted on surface application of slag _ E, lime mud _ Lcal, centrifuged sewage sludge _ LC, dolomitic

  2. D-TRP(8-γMSH Prevents the Effects of Endotoxin in Rat Skeletal Muscle Cells through TNFα/NF-KB Signalling Pathway.

    Directory of Open Access Journals (Sweden)

    Ana Belén Gómez-SanMiguel

    Full Text Available Sepsis induces anorexia and muscle wasting secondary to an increase in muscle proteolysis. Melanocyte stimulating hormones (MSH is a family of peptides that have potent anti-inflammatory effects. Melanocortin receptor-3 (MC3-R has been reported as the predominant anti-inflammatory receptor for melanocortins. The aim of this work was to analyse whether activation of MC3-R, by administration of its agonist D-Trp(8-γMSH, is able to modify the response of skeletal muscle to inflammation induced by lipopolysaccharide endotoxin (LPS or TNFα. Adult male rats were injected with 250 μg/kg LPS and/or 500 μg/kg D-Trp(8-γMSH 17:00 h and at 8:00 h the following day, and euthanized 4 hours afterwards. D-Trp(8-γMSH decreased LPS-induced anorexia and prevented the stimulatory effect of LPS on hypothalamic IL-1β, COX-2 and CRH as well as on serum ACTH and corticosterone. Serum IGF-I and its expression in liver and gastrocnemius were decreased in rats injected with LPS, but not in those that also received D-Trp(8-γMSH. However, D-Trp(8-γMSH was unable to modify the effect of LPS on IGFBP-3. In the gastrocnemius D-Trp(8-γMSH blocked LPS-induced decrease in pAkt, pmTOR, MHC I and MCH II, as well as the increase in pNF-κB(p65, FoxO1, FoxO3, LC3b, Bnip-3, Gabarap1, atrogin-1, MuRF1 and in LC3a/b lipidation. In L6 myotube cultures, D-Trp(8-γMSH was able to prevent TNFα-induced increase of NF-κB(p65 phosphorylation and decrease of Akt phosphorylation as well as of IGF-I and MHC I expression. These data suggest that MC3-R activation prevents the effect of endotoxin on skeletal wasting by modifying inflammation, corticosterone and IGF-I responses and also by directly acting on muscle cells through the TNFα/NF-κB(p65 pathway.

  3. Residual stresses and durability in cold drawn eutectoid steel wires

    Science.gov (United States)

    Atienza, J. M.; Elices, M.; Ruiz-Hervias, J.; Caballero, L.; Valiente, A.

    2007-04-01

    Prestressing steel wires have excellent mechanical properties but there is a need to improve their durability in aggressive environments. In this work, the influence of residual stresses on the environmentally assisted cracking of these wires is studied. A good correlation has been found between residual stresses at the surface of the wires and the time to rupture during stress corrosion test proposed by the International Federation of Prestressing. Wires with the same microstructure, surface quality and mechanical properties show very different behaviour in aggressive environments depending on their residual stress state. Research shows that environmentally assisted cracking can be improved significantly by acting on the surface residual stresses produced by wire drawing. In addition, in this study a post-drawing treatment to generate compressive residual stresses at the surface of the wires is proposed.

  4. Amino Acid Residues Critical for the Specificity for Betaine Aldehyde of the Plant ALDH10 Isoenzyme Involved in the Synthesis of Glycine Betaine1[W][OA

    Science.gov (United States)

    Díaz-Sánchez, Ángel G.; González-Segura, Lilian; Mújica-Jiménez, Carlos; Rudiño-Piñera, Enrique; Montiel, Carmina; Martínez-Castilla, León P.; Muñoz-Clares, Rosario A.

    2012-01-01

    Plant Aldehyde Dehydrogenase10 (ALDH10) enzymes catalyze the oxidation of ω-primary or ω-quaternary aminoaldehydes, but, intriguingly, only some of them, such as the spinach (Spinacia oleracea) betaine aldehyde dehydrogenase (SoBADH), efficiently oxidize betaine aldehyde (BAL) forming the osmoprotectant glycine betaine (GB), which confers tolerance to osmotic stress. The crystal structure of SoBADH reported here shows tyrosine (Tyr)-160, tryptophan (Trp)-167, Trp-285, and Trp-456 in an arrangement suitable for cation-π interactions with the trimethylammonium group of BAL. Mutation of these residues to alanine (Ala) resulted in significant Km(BAL) increases and Vmax/Km(BAL) decreases, particularly in the Y160A mutant. Tyr-160 and Trp-456, strictly conserved in plant ALDH10s, form a pocket where the bulky trimethylammonium group binds. This space is reduced in ALDH10s with low BADH activity, because an isoleucine (Ile) pushes the Trp against the Tyr. Those with high BADH activity instead have Ala (Ala-441 in SoBADH) or cysteine, which allow enough room for binding of BAL. Accordingly, the mutation A441I decreased the Vmax/Km(BAL) of SoBADH approximately 200 times, while the mutation A441C had no effect. The kinetics with other ω-aminoaldehydes were not affected in the A441I or A441C mutant, demonstrating that the existence of an Ile in the second sphere of interaction of the aldehyde is critical for discriminating against BAL in some plant ALDH10s. A survey of the known sequences indicates that plants have two ALDH10 isoenzymes: those known to be GB accumulators have a high-BAL-affinity isoenzyme with Ala or cysteine in this critical position, while non GB accumulators have low-BAL-affinity isoenzymes containing Ile. Therefore, BADH activity appears to restrict GB synthesis in non-GB-accumulator plants. PMID:22345508

  5. Histidine 352 (His352 and tryptophan 355 (Trp355 are essential for flax UGT74S1 glucosylation activity toward secoisolariciresinol.

    Directory of Open Access Journals (Sweden)

    Kaushik Ghose

    Full Text Available Flax secoisolariciresinol diglucoside (SDG lignan is a natural phytoestrogen for which a positive role in metabolic diseases is emerging. Until recently however, much less was known about SDG and its monoglucoside (SMG biosynthesis. Lately, flax UGT74S1 was identified and characterized as an enzyme sequentially glucosylating secoisolariciresinol (SECO into SMG and SDG when expressed in yeast. However, the amino acids critical for UGT74S1 glucosyltransferase activity were unknown. A 3D structural modeling and docking, site-directed mutagenesis of five amino acids in the plant secondary product glycosyltransferase (PSPG motif, and enzyme assays were conducted. UGT74S1 appeared to be structurally similar to the Arabidopsis thaliana UGT72B1 model. The ligand docking predicted Ser357 and Trp355 as binding to the phosphate and hydroxyl groups of UDP-glucose, whereas Cys335, Gln337 and Trp355 were predicted to bind the 7-OH, 2-OCH3 and 17-OCH3 of SECO. Site-directed mutagenesis of Cys335, Gln337, His352, Trp355 and Ser357, and enzyme assays revealed an alteration of these binding sites and a significant reduction of UGT74S1 glucosyltransferase catalytic activity towards SECO and UDP-glucose in all mutants. A complete abolition of UGT74S1 activity was observed when Trp355 was substituted to Ala355 and Gly355 or when changing His352 to Asp352, and an altered metabolite profile was observed in Cys335Ala, Gln337Ala, and Ser357Ala mutants. This study provided for the first time evidence that Trp355 and His352 are critical for UGT74S1's glucosylation activity toward SECO and suggested the possibility for SMG production in vitro.

  6. Impaired 8-Hydroxyguanine Repair Activity of MUTYH Variant p.Arg109Trp Found in a Japanese Patient with Early-Onset Colorectal Cancer

    Directory of Open Access Journals (Sweden)

    Kazuya Shinmura

    2014-01-01

    Full Text Available Purpose. The biallelic inactivation of the 8-hydroxyguanine repair gene MUTYH leads to MUTYH-associated polyposis (MAP, which is characterized by colorectal multiple polyps and carcinoma(s. However, only limited information regarding MAP in the Japanese population is presently available. Since early-onset colorectal cancer (CRC is a characteristic of MAP and might be caused by the inactivation of another 8-hydroxyguanine repair gene, OGG1, we investigated whether germline MUTYH and OGG1 mutations are involved in early-onset CRC in Japanese patients. Methods. Thirty-four Japanese patients with early-onset CRC were examined for germline MUTYH and OGG1 mutations using sequencing. Results. Biallelic pathogenic mutations were not found in any of the patients; however, a heterozygous p.Arg19*  MUTYH variant and a heterozygous p.Arg109Trp MUTYH variant were detected in one patient each. The p.Arg19* and p.Arg109Trp corresponded to p.Arg5* and p.Arg81Trp, respectively, in the type 2 nuclear-form protein. The defective DNA repair activity of p.Arg5* is apparent, while that of p.Arg81Trp has been demonstrated using DNA cleavage and supF forward mutation assays. Conclusion. These results suggest that biallelic MUTYH or OGG1 pathogen