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Sample records for metal mediated transformations

  1. [Transition metal mediated transformations of small molecules

    SciTech Connect

    Sen, A.

    1992-01-01

    Work on organotransition metal chemistry, homogeneous and heterogeneous catalysis is summarized. Several cationic palladium(II) complexes with bulky phosphine or pyridine ligands were discovered that are highly selective catalysts for linear dimerization of vinyl monomers and linear polymerization of p-divinylbenzene, the reactions proceeding through a carbocationic mechanism. Our studies were continued on alternating olefin-carbon monoxide copolymers. The copolymerization reaction and reactivity of copolymers were examined. New catalytic systems for alternating copolymerization of [alpha]-olefins with CO were discovered. In the case of styrene derivatives, tactic copolymers were obtained. Poly(ethylenepyrrolediyl) derivatives were synthesized from alternating ethylene-carbon monoxide copolymer and become electronic conductors when doped with iodine. A catalytic system for direct synthesis of polyureas and polyoxamides from and diamines was also discovered. Pt metal catalyzed the oxidation of ethers, esters, and amines to carboxylic acids and the oxidation of olefins to 1,2-diols. Anaerobic and aerobic decomposition of molybdenum(VI)-oxoalkyl compounds were studied for heterogeneous oxidation of alkanes and olefins on Mo(VI)-oxide surfaces. Synthesis of polymer-trapped metal, metal oxide, and metal sulfide nanoclusters (size <1--10 nm) was studied.

  2. [Transition metal mediated transformations of small molecules]. Progress report

    SciTech Connect

    Sen, A.

    1992-10-01

    Work on organotransition metal chemistry, homogeneous and heterogeneous catalysis is summarized. Several cationic palladium(II) complexes with bulky phosphine or pyridine ligands were discovered that are highly selective catalysts for linear dimerization of vinyl monomers and linear polymerization of p-divinylbenzene, the reactions proceeding through a carbocationic mechanism. Our studies were continued on alternating olefin-carbon monoxide copolymers. The copolymerization reaction and reactivity of copolymers were examined. New catalytic systems for alternating copolymerization of {alpha}-olefins with CO were discovered. In the case of styrene derivatives, tactic copolymers were obtained. Poly(ethylenepyrrolediyl) derivatives were synthesized from alternating ethylene-carbon monoxide copolymer and become electronic conductors when doped with iodine. A catalytic system for direct synthesis of polyureas and polyoxamides from and diamines was also discovered. Pt metal catalyzed the oxidation of ethers, esters, and amines to carboxylic acids and the oxidation of olefins to 1,2-diols. Anaerobic and aerobic decomposition of molybdenum(VI)-oxoalkyl compounds were studied for heterogeneous oxidation of alkanes and olefins on Mo(VI)-oxide surfaces. Synthesis of polymer-trapped metal, metal oxide, and metal sulfide nanoclusters (size <1--10 nm) was studied.

  3. Main Group Lewis Acid-Mediated Transformations of Transition-Metal Hydride Complexes.

    PubMed

    Maity, Ayan; Teets, Thomas S

    2016-08-10

    This Review highlights stoichiometric reactions and elementary steps of catalytic reactions involving cooperative participation of transition-metal hydrides and main group Lewis acids. Included are reactions where the transition-metal hydride acts as a reactant as well as transformations that form the metal hydride as a product. This Review is divided by reaction type, illustrating the diverse roles that Lewis acids can play in mediating transformations involving transition-metal hydrides as either reactants or products. We begin with a discussion of reactions where metal hydrides form direct adducts with Lewis acids, elaborating the structure and dynamics of the products of these reactions. The bulk of this Review focuses on reactions where the transition metal and Lewis acid act in cooperation, and includes sections on carbonyl reduction, H2 activation, and hydride elimination reactions, all of which can be promoted by Lewis acids. Also included is a section on Lewis acid-base secondary coordination sphere interactions, which can influence the reactivity of hydrides. Work from the past 50 years is included, but the majority of this Review focuses on research from the past decade, with the intent of showcasing the rapid emergence of this field and the potential for further development into the future.

  4. Laccase-mediated transformation of triclosan in aqueous solution with metal cations and humic acid.

    PubMed

    Sun, Kai; Kang, Fuxing; Waigi, Michael Gatheru; Gao, Yanzheng; Huang, Qingguo

    2017-01-01

    Triclosan (TCS) is a broad-spectrum antimicrobial agent that is found extensively in natural aquatic environments. Enzyme-catalyzed oxidative coupling reactions (ECOCRs) can be used to remove TCS in aqueous solution, but there is limited information available to indicate how metal cations (MCs) and natural organic matter (NOM) influence the environmental fate of TCS during laccase-mediated ECOCRs. In this study, we demonstrated that the naturally occurring laccase from Pleurotus ostreatus was effective in removing TCS during ECOCRs, and the oligomerization of TCS was identified as the dominant reaction pathway by high-resolution mass spectrometry (HRMS). The growth inhibition studies of green algae (Chlamydomonas reinhardtii and Scenedesmus obliquus) proved that laccase-mediated ECOCRs could effectively reduce the toxicity of TCS. The presence of dissolved MCs (Mn(2+), Al(3+), Ca(2+), Cu(2+), and Fe(2+) ions) influenced the removal and transformation of TCS via different mechanisms. Additionally, the transformation of TCS in systems with NOM derived from humic acid (HA) was hindered, and the apparent pseudo first-order kinetics rate constants (k) for TCS decreased as the HA concentration increased, which likely corresponded to the combined effect of both noncovalent (sorption) and covalent binding between TCS and humic molecules. Our results provide a novel insight into the fate and transformation of TCS by laccase-mediated ECOCRs in natural aquatic environments in the presence of MCs and NOM. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Nickel and cadmium-induced SLBP depletion: A potential pathway to metal mediated cellular transformation

    PubMed Central

    Jordan, Ashley; Zhang, Xiaoru; Li, Jinquan; Laulicht-Glick, Freda; Sun, Hong

    2017-01-01

    Both nickel and cadmium compounds have been established as group I carcinogens for several decades. Despite over-whelming evidence of these compounds’ carcinogenicity in humans, the specific underlying molecular mechanisms that govern metal induced cellular transformation remain unclear. In this study, we found that there were slightly different effects on decreased SLBP mRNA and protein as well as increased polyA H3.1 in our nickel exposed cells. This suggested that nickel and arsenic have similar effects on canonical histone mRNA transcription and translation. We also saw that the depletion of SLBP protein was reversed by inhibiting the proteosome. Finally, we showed that inhibiting the SLBP mRNA and protein levels were rescued by epigenetic modifiers suggesting that nickel’s effects on SLBP may be mediated via epigenetic mechanisms. Taken together these results suggest a similar mechanism by which both arsenic and nickel may exert their carcinogenic effects. PMID:28306745

  6. Solvent-Mediated Transformation from Achiral to Chiral Nickel(II) Metal-Organic Frameworks and Reassembly in Solution.

    PubMed

    Li, Xiaoju; Yu, Zhenjiang; Li, Xinxiong; Guo, Xiaofang

    2015-11-09

    Reactions of 5-nitroisophthalic acid (NO2 -H2 ip), 1,4-bis(imidazol-1'-yl)butane (bimb), and Ni(NO3 )2 ⋅6 H2 O gave rise to four metal-organic frameworks (MOFs), [Ni2 (NO2 -ip)2 (bimb)1.5 ]n (1), [Ni4 (NO2 -ip)3 (bimb)2 (OH)2 (H2 O)]n ⋅(CH3 CH2 OH)0.5 n (2), [Ni(NO2 -ip)(bimb)1.5 (H2 O)]n ⋅(H2 O)n ⋅(CH3 CH2 OH)0.5 n (3), and [Ni(NO2 -ip) (bimb)(μ-H2 O)]n ⋅(H2 O)n (4). The metal/ligand ratio, pH value, and solvent exerted a subtle but crucial influence on the formation of complexes 1-4, which possess different visual color and crystal structures. Complex 1 exhibits a twofold interpenetrating 3D pillared bilayer framework composed of binuclear and mononuclear Ni(II) units, whereas complex 2 is a 3D chiral network that consists of asymmetric tetranuclear Ni(II) units. Complexes 3 and 4 are 3D layer-pillared frameworks that consist of mononuclear Ni(II) ions and a 3D six-connected network of μ-water-bridged dinuclear Ni(II) units, respectively. Interestingly, achiral 4 can be transformed into chiral 2 by using a solvent-mediated single-crystal-to-single-crystal process without any chiral auxiliary. Magnetic analyses of 2 and 4 show the occurrence of antiferromagnetic interactions. Complex 3 is difficult to obtain directly as a single solid phase, but it can be homogeneously formed by solvent-mediated transformations from 1, 2, and 4. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Metal-Organic Nanosheets Formed via Defect-Mediated Transformation of a Hafnium Metal-Organic Framework.

    PubMed

    Cliffe, Matthew J; Castillo-Martínez, Elizabeth; Wu, Yue; Lee, Jeongjae; Forse, Alexander C; Firth, Francesca C N; Moghadam, Peyman Z; Fairen-Jimenez, David; Gaultois, Michael W; Hill, Joshua A; Magdysyuk, Oxana V; Slater, Ben; Goodwin, Andrew L; Grey, Clare P

    2017-04-19

    We report a hafnium-containing MOF, hcp UiO-67(Hf), which is a ligand-deficient layered analogue of the face-centered cubic fcu UiO-67(Hf). hcp UiO-67 accommodates its lower ligand:metal ratio compared to fcu UiO-67 through a new structural mechanism: the formation of a condensed "double cluster" (Hf12O8(OH)14), analogous to the condensation of coordination polyhedra in oxide frameworks. In oxide frameworks, variable stoichiometry can lead to more complex defect structures, e.g., crystallographic shear planes or modules with differing compositions, which can be the source of further chemical reactivity; likewise, the layered hcp UiO-67 can react further to reversibly form a two-dimensional metal-organic framework, hxl UiO-67. Both three-dimensional hcp UiO-67 and two-dimensional hxl UiO-67 can be delaminated to form metal-organic nanosheets. Delamination of hcp UiO-67 occurs through the cleavage of strong hafnium-carboxylate bonds and is effected under mild conditions, suggesting that defect-ordered MOFs could be a productive route to porous two-dimensional materials.

  8. Metal-mediated oxidation of fluoroquinolone antibiotics in water: A review on kinetics, transformation products, and toxicity assessment.

    PubMed

    Feng, Mingbao; Wang, Zunyao; Dionysiou, Dionysios D; Sharma, Virender K

    2017-08-30

    Fluoroquinolones (FQs) are among the most potent antimicrobial agents, which have seen their increasing use as human and veterinary medicines to control bacterial infections. FQs have been extensively found in surface water and municipal wastewaters, which has raised great concerns due to their negative impacts to humans and ecological health. It is of utmost importance that FQs are treated before their release into the environment. This paper reviews oxidative removal of FQs using reactive oxygen (O3 and OH), sulfate radicals (SO4(-)), and high-valent transition metal (Mn(VII) and Fe(VI)) species. The role of metals in enhancing the performance of reactive oxygen and sulfur species is presented. The catalysts can significantly enhance the production of OH and/or SO4(-) radicals. At neutral pH, the second-order rate constants (k, M(-1)s(-1)) of the reactions between FQs and oxidants follow the order as k(OH)>k(O3)>k(Fe(VI))>k(Mn(VII)). Moieties involved to transform target FQs to oxidized products and participation of the catalysts in the reaction pathways are discussed. Generally, the piperazinyl ring of FQs was found as the preferential attack site by each oxidant. Meanwhile, evaluation of aquatic ecotoxicity of the transformation products of FQs by these treatments is summarized. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Transformable liquid-metal nanomedicine

    PubMed Central

    Lu, Yue; Hu, Quanyin; Lin, Yiliang; Pacardo, Dennis B.; Wang, Chao; Sun, Wujin; Ligler, Frances S.; Dickey, Michael D.; Gu, Zhen

    2015-01-01

    To date, numerous inorganic nanocarriers have been explored for drug delivery systems (DDSs). However, the clinical application of inorganic formulations has often been hindered by their toxicity and failure to biodegrade. We describe here a transformable liquid-metal nanomedicine, based on a core–shell nanosphere composed of a liquid-phase eutectic gallium-indium core and a thiolated polymeric shell. This formulation can be simply produced through a sonication-mediated method with bioconjugation flexibility. The resulting nanoparticles loaded with doxorubicin (Dox) have an average diameter of 107 nm and demonstrate the capability to fuse and subsequently degrade under a mildly acidic condition, which facilitates release of Dox in acidic endosomes after cellular internalization. Equipped with hyaluronic acid, a tumour-targeting ligand, this formulation displays enhanced chemotherapeutic inhibition towards the xenograft tumour-bearing mice. This liquid metal-based DDS with fusible and degradable behaviour under physiological conditions provides a new strategy for engineering theranostic agents with low toxicity. PMID:26625944

  10. Transformable liquid-metal nanomedicine

    NASA Astrophysics Data System (ADS)

    Lu, Yue; Hu, Quanyin; Lin, Yiliang; Pacardo, Dennis B.; Wang, Chao; Sun, Wujin; Ligler, Frances S.; Dickey, Michael D.; Gu, Zhen

    2015-12-01

    To date, numerous inorganic nanocarriers have been explored for drug delivery systems (DDSs). However, the clinical application of inorganic formulations has often been hindered by their toxicity and failure to biodegrade. We describe here a transformable liquid-metal nanomedicine, based on a core-shell nanosphere composed of a liquid-phase eutectic gallium-indium core and a thiolated polymeric shell. This formulation can be simply produced through a sonication-mediated method with bioconjugation flexibility. The resulting nanoparticles loaded with doxorubicin (Dox) have an average diameter of 107 nm and demonstrate the capability to fuse and subsequently degrade under a mildly acidic condition, which facilitates release of Dox in acidic endosomes after cellular internalization. Equipped with hyaluronic acid, a tumour-targeting ligand, this formulation displays enhanced chemotherapeutic inhibition towards the xenograft tumour-bearing mice. This liquid metal-based DDS with fusible and degradable behaviour under physiological conditions provides a new strategy for engineering theranostic agents with low toxicity.

  11. Co-metal-organic-frameworks with pure uniform crystal morphology prepared via Co2 + exchange-mediated transformation from Zn-metallogels for luminol catalysed chemiluminescence

    NASA Astrophysics Data System (ADS)

    Tang, Xue Qian; Xiao, Bo Wen; Li, Chun Mei; Wang, Dong Mei; Huang, Cheng Zhi; Li, Yuan Fang

    2017-03-01

    Cation exchange-mediated transformation from Zn-metallogels (MOGs), which was a mild facile strategy relative to the demanding hydrothermal method, was employed to develop Co2 + metal-organic frameworks (Co-MOFs) at room temperature. The obtained Co-MOFs was of uniform octahedral morphology and possessed high activity to catalyze luminol chemiluminescence without extra oxidants. By adding cysteine, the CL emission of luminol-Co-MOFs system was further enhanced. Based on this phenomenon, Co-MOFs was utilized to build a practical sensing platform for cysteine determination. Under the optimized conditions, the relative CL intensity (ΔI) was proportional to the concentration of cysteine in the range of 2-10 μM, and the detection limit was 0.49 μM (3S/N). Moreover, the established method was applied to the determination of cysteine in commercially available pharmaceutical injections.

  12. Advances in Stereoconvergent Catalysis from 2005 to 2015: Transition-Metal-Mediated Stereoablative Reactions, Dynamic Kinetic Resolutions, and Dynamic Kinetic Asymmetric Transformations.

    PubMed

    Bhat, Vikram; Welin, Eric R; Guo, Xuelei; Stoltz, Brian M

    2017-03-08

    Stereoconvergent catalysis is an important subset of asymmetric synthesis that encompasses stereoablative transformations, dynamic kinetic resolutions, and dynamic kinetic asymmetric transformations. Initially, only enzymes were known to catalyze dynamic kinetic processes, but recently various synthetic catalysts have been developed. This Review summarizes major advances in nonenzymatic, transition-metal-promoted dynamic asymmetric transformations reported between 2005 and 2015.

  13. Agrobacterium-mediated sorghum transformation.

    PubMed

    Zhao, Z Y; Cai, T; Tagliani, L; Miller, M; Wang, N; Pang, H; Rudert, M; Schroeder, S; Hondred, D; Seltzer, J; Pierce, D

    2000-12-01

    Agrobacterium tumefaciens was used to genetically transform sorghum. Immature embryos of a public (P898012) and a commercial line (PHI391) of sorghum were used as the target explants. The Agrobacterium strain used was LBA4404 carrying a 'Super-binary' vector with a bar gene as a selectable marker for herbicide resistance in the plant cells. A series of parameter tests was used to establish a baseline for conditions to be used in stable transformation experiments. A number of different transformation conditions were tested and a total of 131 stably transformed events were produced from 6175 embryos in these two sorghum lines. Statistical analysis showed that the source of the embryos had a very significant impact on transformation efficiency, with field-grown embryos producing a higher transformation frequency than greenhouse-grown embryos. Southern blot analysis of DNA from leaf tissues of T0 plants confirmed the integration of the T-DNA into the sorghum genome. Mendelian segregation in the T1 generation was confirmed by herbicide resistance screening. This is the first report of successful use of Agrobacterium for production of stably transformed sorghum plants. The Agrobacterium method we used yields a higher frequency of stable transformation that other methods reported previously.

  14. Barley Transformation Using Agrobacterium-Mediated Techniques

    NASA Astrophysics Data System (ADS)

    Harwood, Wendy A.; Bartlett, Joanne G.; Alves, Silvia C.; Perry, Matthew; Smedley, Mark A.; Leyland, Nicola; Snape, John W.

    Methods for the transformation of barley using Agrobacterium-mediated techniques have been available for the past 10 years. Agrobacterium offers a number of advantages over biolistic-mediated techniques in terms of efficiency and the quality of the transformed plants produced. This chapter describes a simple system for the transformation of barley based on the infection of immature embryos with Agrobacterium tumefaciens followed by the selection of transgenic tissue on media containing the antibiotic hygromycin. The method can lead to the production of large numbers of fertile, independent transgenic lines. It is therefore ideal for studies of gene function in a cereal crop system.

  15. Metal Ions Mediated Morphology and Phase Transformation of Chalcogenide Semiconductor: From CuClSe2 Microribbon to CuSe Nanosheet.

    PubMed

    Liu, Yong-Qiang; Wu, Hao-Di; Zhao, Yu; Pan, Ge-Bo

    2015-05-05

    Foreign ions are of significant importance in controlling and modulating the morphology of semiconductor nanocrystals during the colloidal synthesis process. Herein, we demonstrate the potential of foreign metal ions to simultaneously control the morphology and crystal phase of chalcogenide semiconductors. The results indicate that the introduction of Al(3+) ions can induce the structural transformation from monoclinic CuClSe2 microribbons (MRs) to klockmannite CuSe nanosheets (NSs) and the growth of large-sized CuSe NSs. The as-prepared micrometer-sized CuSe NSs exhibit a high-conducting behavior, long-term durability, and environment stability. The novel properties enable CuSe NSs to open up a bright prospect for printable electrical interconnects and flexible electronic devices.

  16. Microbial mediated retention/transformation of organic and inorganic materials in freshwater and marine ecosystems

    EPA Science Inventory

    Aquatic ecosystems are globally connected by hydrological and biogeochemical cycles. Microorganisms inhabiting aquatic ecosystems form the basis of food webs, mediate essential element cycles, decompose natural organic matter, transform inorganic nutrients and metals, and degrad...

  17. Microbial mediated retention/transformation of organic and inorganic materials in freshwater and marine ecosystems

    EPA Science Inventory

    Aquatic ecosystems are globally connected by hydrological and biogeochemical cycles. Microorganisms inhabiting aquatic ecosystems form the basis of food webs, mediate essential element cycles, decompose natural organic matter, transform inorganic nutrients and metals, and degrad...

  18. Rh(I)-catalyzed transformation of propargyl vinyl ethers into (E,Z)-dienals: stereoelectronic role of trans effect in a metal-mediated pericyclic process and a shift from homogeneous to heterogeneous catalysis during a one-pot reaction.

    PubMed

    Vidhani, Dinesh V; Krafft, Marie E; Alabugin, Igor V

    2014-01-03

    The combination of experiments and computations reveals unusual features of stereoselective Rh(I)-catalyzed transformation of propargyl vinyl ethers into (E,Z)-dienals. The first step, the conversion of propargyl vinyl ethers into allene aldehydes, proceeds under homogeneous conditions via a "cyclization-mediated" mechanism initiated by Rh(I) coordination at the alkyne. This path agrees well with the small experimental effects of substituents on the carbinol carbon. The key feature revealed by the computational study is the stereoelectronic effect of the ligand arrangement at the catalytic center. The rearrangement barriers significantly decrease due to the greater transfer of electron density from the catalytic metal center to the CO ligand oriented trans to the alkyne. This effect increases electrophilicity of the metal and lowers the calculated barriers by 9.0 kcal/mol. Subsequent evolution of the catalyst leads to the in situ formation of Rh(I) nanoclusters that catalyze stereoselective tautomerization. The intermediacy of heterogeneous catalysis by nanoclusters was confirmed by mercury poisoning, temperature-dependent sigmoidal kinetic curves, and dynamic light scattering. The combination of experiments and computations suggests that the initially formed allene-aldehyde product assists in the transformation of a homogeneous catalyst (or "a cocktail of catalysts") into nanoclusters, which in turn catalyze and control the stereochemistry of subsequent transformations.

  19. Agrobacterium-mediated transformation of Fusarium proliferatum.

    PubMed

    Bernardi-Wenzel, J; Quecine, M C; Azevedo, J L; Pamphile, J A

    2016-06-03

    Fusarium proliferatum is an important pathogen that is associated with plant diseases and primarily affects aerial plant parts by producing different mycotoxins, which are toxic to humans and animals. Within the last decade, this fungus has also been described as one of the causes of red root rot or sudden death syndrome in soybean, which causes extensive damage to this crop. This study describes the Agrobacterium tumefaciens-mediated transformation of F. proliferatum as a tool for the disruption of pathogenicity genes. The genetic transformation was performed using two binary vectors (pCAMDsRed and pFAT-GFP) containing the hph (hygromycin B resistance) gene as a selection marker and red and green fluorescence, respectively. The presence of acetosyringone and the use of filter paper or nitrocellulose membrane were evaluated for their effect on the transformation efficiency. A mean processing rate of 94% was obtained with 96 h of co-cultivation only in the presence of acetosyringone and the use of filter paper or nitrocellulose membrane did not affect the transformation process. Hygromycin B resistance and the presence of the hph gene were confirmed by PCR, and fluorescence due to the expression of GFP and DsRed protein was monitored in the transformants. A high rate of mitotic stability (95%) was observed. The efficiency of Agrobacterium-mediated transformation of F. proliferatum allows the technique to be used for random insertional mutagenesis studies and to analyze fungal genes involved in the infection process.

  20. Complexion-mediated martensitic phase transformation in Titanium

    NASA Astrophysics Data System (ADS)

    Zhang, J.; Tasan, C. C.; Lai, M. J.; Dippel, A.-C.; Raabe, D.

    2017-02-01

    The most efficient way to tune microstructures and mechanical properties of metallic alloys lies in designing and using athermal phase transformations. Examples are shape memory alloys and high strength steels, which together stand for 1,500 million tons annual production. In these materials, martensite formation and mechanical twinning are tuned via composition adjustment for realizing complex microstructures and beneficial mechanical properties. Here we report a new phase transformation that has the potential to widen the application window of Ti alloys, the most important structural material in aerospace design, by nanostructuring them via complexion-mediated transformation. This is a reversible martensitic transformation mechanism that leads to a final nanolaminate structure of α'' (orthorhombic) martensite bounded with planar complexions of athermal ω (a-ω, hexagonal). Both phases are crystallographically related to the parent β (BCC) matrix. As expected from a planar complexion, the a-ω is stable only at the hetero-interface.

  1. Complexion-mediated martensitic phase transformation in Titanium.

    PubMed

    Zhang, J; Tasan, C C; Lai, M J; Dippel, A-C; Raabe, D

    2017-02-01

    The most efficient way to tune microstructures and mechanical properties of metallic alloys lies in designing and using athermal phase transformations. Examples are shape memory alloys and high strength steels, which together stand for 1,500 million tons annual production. In these materials, martensite formation and mechanical twinning are tuned via composition adjustment for realizing complex microstructures and beneficial mechanical properties. Here we report a new phase transformation that has the potential to widen the application window of Ti alloys, the most important structural material in aerospace design, by nanostructuring them via complexion-mediated transformation. This is a reversible martensitic transformation mechanism that leads to a final nanolaminate structure of α″ (orthorhombic) martensite bounded with planar complexions of athermal ω (a-ω, hexagonal). Both phases are crystallographically related to the parent β (BCC) matrix. As expected from a planar complexion, the a-ω is stable only at the hetero-interface.

  2. Complexion-mediated martensitic phase transformation in Titanium

    PubMed Central

    Zhang, J.; Tasan, C. C.; Lai, M. J.; Dippel, A. -C.; Raabe, D.

    2017-01-01

    The most efficient way to tune microstructures and mechanical properties of metallic alloys lies in designing and using athermal phase transformations. Examples are shape memory alloys and high strength steels, which together stand for 1,500 million tons annual production. In these materials, martensite formation and mechanical twinning are tuned via composition adjustment for realizing complex microstructures and beneficial mechanical properties. Here we report a new phase transformation that has the potential to widen the application window of Ti alloys, the most important structural material in aerospace design, by nanostructuring them via complexion-mediated transformation. This is a reversible martensitic transformation mechanism that leads to a final nanolaminate structure of α″ (orthorhombic) martensite bounded with planar complexions of athermal ω (a–ω, hexagonal). Both phases are crystallographically related to the parent β (BCC) matrix. As expected from a planar complexion, the a–ω is stable only at the hetero-interface. PMID:28145484

  3. Anomalous plasticity in defect-mediated phase transformations

    NASA Astrophysics Data System (ADS)

    Ghimire, Punam; Ravelo, R.; Germann, T. C.

    2014-03-01

    Large-scale molecular dynamics simulations of shocked wave propagation in metallic single crystals exhibit high elastic limits and are ideally suited for investigating the role defect nucleation and multiplication play on the kinetics of phase transformations. Here we report on the morphology and kinetics of shocked-induced phase transformations in Aluminum single crystals. The atomic interactions were modeled utilizing various embedded atom method (EAM) models of Aluminum, with most models exhibiting an artificial fcc -->bcc phase transformation in the 25-30 GPa range. For cases where plastic deformation precedes the phase transformation, anomalous defect structures atypical of plastic deformation in bcc lattices nucleate early on but anneal out with time. In all cases, the defect-mediated phase transitions proceed at faster rates than defect-free ones. Part of this work supported by the Air Force Office of Scientific Research under AFOSR Award No. FA9550-12-1-0476.

  4. Can microbes mediate nano-transformation?

    NASA Astrophysics Data System (ADS)

    Prasad, K.; Jha, Anal K.; Prasad, Kamlesh; Kulkarni, A. R.

    2010-10-01

    A green low-cost and reproducible microbe ( Lactobacillus) and baker's yeast ( Saccharomyces cerevisiae) mediated biosynthesis of metallic and oxide nanoparticles are reported. Silver and copper oxide nanoparticles are synthesized using Lactobacillus sp. and Saccharomyces cerevisiae. The synthesis is performed akin to room temperature in the laboratory ambience. X-ray and transmission electron microscopy analyses are performed to ascertain the formation of metallic and oxide nanoparticles. Individual nanoparticles having the dimensions of 2-6 nm (metallic) and 10-20 nm (oxide) are found. A possible mechanism involved for the synthesis of metallic and oxide nanoparticles has also been proposed in which pH as well as the partial pressure of gaseous hydrogen (r-H2) or redox potential of the culture solution seem to play an important role in the process.

  5. Trace metal transformations in gasification

    SciTech Connect

    Erickson, T.A.; Zygarlicke, C.J.; O`Keefe, C.A.

    1995-08-01

    The Energy & Environmental Research Center (EERC) is carrying out an investigation that will provide methods to predict the fate of selected trace elements in integrated gasification combined cycle (IGCC) and integrated gasification fuel cell (IGFC) systems to aid in the development of methods to control the emission of trace elements determined to be air toxics. The goal of this project is to identify the effects of critical chemical and physical transformations associated with trace element behavior in IGCC and IGFC systems. The trace elements included in this project are arsenic, chromium, cadmium, mercury, nickel, selenium, and lead. The research seeks to identify and fill, experimentally and/or theoretically, data gaps that currently exist on the fate and composition of trace elements. The specific objectives are to (1) review the existing literature to identify the type and quantity of trace elements from coal gasification systems, (2) perform laboratory-scale experimentation and computer modeling to enable prediction of trace element emissions, and (3) identify methods to control trace element emissions.

  6. Structure and phase transformations in uranium metal

    NASA Astrophysics Data System (ADS)

    Axe, J. D.; Gruebel, G.; Lander, G. H.

    1994-10-01

    In common with other elemental actinides, metallic uranium exists in several allotropic forms, differing from one another by complex but subtle atomic rearrangements. This article reviews progress in understanding the successive phase transformations from the perspective of soft-mode instabilities.

  7. Structure and phase transformations in uranium metal

    SciTech Connect

    Axe, J.D.; Gruebel, G.; Lander, G.H.

    1993-12-01

    In common with other elemental actinides, metallic uranium exists in several allotropic forms, differing from one another by complex but subtle atomic rearrangements. This article reviews progress in understanding the successive phase transformations from the perspective of soft-mode instabilities.

  8. Transition metal catalyzed transformations of unsaturated molecules

    SciTech Connect

    Not Available

    1989-01-01

    In this proposal, research in three areas of transition metal catalyzed transformations of small molecules is proposed. The first encompasses metal catalyzed processes for the synthesis of several classes of carbon monoxide containing polymers. This section describes plans for metal catalyzed synthesis of (a) new alternating copolymers of carbon monoxide and olefins, (b) block copolymers consisting of segments of the olefin homopolymer and the olefin- carbon monoxide alternating copolymer, and (c) polycarbonates, polyesters and polyamides. The second section involves the examination of the chemistry of metal complexes incorporating oxo and hydrocarbyl ligands as a model for the heterogeneous oxidation of olefins and alkanes by meal oxides. Specific plans are to mimic in solution two proposed key steps in the heterogeneous oxidations. These are (a) the heterolytic cleavage of an alkyl (or allyl) C-H bond that is assisted by an oxo group, and (a) the transfer of an oxo group to the resultant metal bound alkyl (or allyl) ligand. The third section concerned with the development of a hybrid catalyst system involving both homogeneous and heterogeneous components for the oxidative functionalization of alkanes. The basic idea is to employ a transition metal in the elemental state to activate C-H bonds of alkanes and form surface alkyl groups. An additional transition metal species will be present in solution which will serve to oxidize these surface alkyl groups to ultimately yield oxidatively functionalized organic products. 57 refs.

  9. Metal-mediated targeting in the body.

    PubMed

    Tang, Xuan; Liang, Xiangyang

    2013-03-01

    Metal ions are important for many biological processes and are steadily available in the human body. Metal concentrations can be extremely high in diseased areas of various pathological conditions. Some synthetic and natural drugs need to be activated by metal ions as prodrugs. In this review, we provide a few examples to illustrate how metal ions activate and mediate drug targeting in the body. This knowledge may be helpful for the development of more effective drugs and pharmaceutical formulations.

  10. Plant transformation via pollen tube-mediated gene transfer

    USDA-ARS?s Scientific Manuscript database

    Genetic transformation using foreign genes and the subsequent development of transgenic plants has been employed to develop enhanced elite germplasm. Although some skepticism exits regarding pollen tube-mediated gene transfer (PTT), reports demonstrating improved transformation efficiency with PTT ...

  11. Avant-garde metalating agents: structural basis of alkali-metal-mediated metalation.

    PubMed

    Mulvey, Robert E

    2009-06-16

    Metalation, one of the most useful and widely used synthetic methodologies, transforms a relatively inert carbon-hydrogen bond to a more labile carbon-metal bond. Until recently, most organometallic reagents that facilitate this process have combined strongly electropositive metals, such as lithium, with organic reagents to form highly polar and, by implication, highly reactive carbon-metal bonds. For example, the alkyllithium reagents and bulky lithium amides that are commonly employed for this purpose can suffer from low functional group tolerance. Lithio-products of these reactions generally have low kinetic stabilities. More recently, several groups around the world have pioneered alternative metalation reagents, complex metalators, which can be interpreted as composite molecules or mixtures made up of two or more distinct compound types. Several examples include magnesiate complexes, Lochmann-Schlosser superbases, Kondo and Uchiyama's 2,2,6,6-tetramethylpiperidide (TMP)-zincate complexes, and Knochel's turbo-Grignard and related salt-supported reagents. This Account describes our rational development of novel complex metalators based on existing structural templates and designed to execute alkali-metal-mediated metalations (AMMMs). By changing the nonalkali metal in these structures, we have produced tailor-made dianionic-dicationic structures such as [(TMEDA).Na(mu-TMP)(mu-(n)Bu)Mg(TMP)], [(TMEDA).Na(mu-TMP)(mu-(t)Bu)Zn((t)Bu)], and [(TMEDA).Li(mu-TMP)Mn(CH(2)SiMe(3))(2)] (TMEDA = N,N,N',N'-tetramethylethylenediamine). These compounds can perform unprecedented magnesiations, zincations, or manganations on aromatic substrates that are generally inert toward conventional Mg, Zn, or Mn(II) reagents. Although the alkali metal is an essential component of these new complex metalators, interestingly, the less electropositive, less polar nonalkali metal [Mg, Zn, or Mn(II)] actually carries out the deprotonation. We view this unique behavior as a mixed-metal synergic

  12. Role of oxidative stress in transformation induced by metal mixture.

    PubMed

    Martín, Silva-Aguilar; Emilio, Rojas; Mahara, Valverde

    2011-01-01

    Metals are ubiquitous pollutants present as mixtures. In particular, mixture of arsenic-cadmium-lead is among the leading toxic agents detected in the environment. These metals have carcinogenic and cell-transforming potential. In this study, we used a two step cell transformation model, to determine the role of oxidative stress in transformation induced by a mixture of arsenic-cadmium-lead. Oxidative damage and antioxidant response were determined. Metal mixture treatment induces the increase of damage markers and the antioxidant response. Loss of cell viability and increased transforming potential were observed during the promotion phase. This finding correlated significantly with generation of reactive oxygen species. Cotreatment with N-acetyl-cysteine induces effect on the transforming capacity; while a diminution was found in initiation, in promotion phase a total block of the transforming capacity was observed. Our results suggest that oxidative stress generated by metal mixture plays an important role only in promotion phase promoting transforming capacity.

  13. Tandem metal-mediated synthesis

    SciTech Connect

    Baker, R.T.

    1998-12-31

    This is the final report of a one-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). Boronic acids RB(OH){sub 2} are currently of interest for applications involving molecular recognition such as amine and sugar sensors and selective transport of biomolecules. They have also been shown to be powerful enzyme inhibitors and alpha-aminoboronic acids (ABAs) are particularly selective inhibitors of serine proteases. Unfortunately, current multistep synthetic routes to the latter compounds do not allow for the incorporation of a wide variety of organic substituents R that may lead to new, more selective enzyme inhibitors. In this work the author describes several direct routes to ABAs based on metal-catalyzed addition of diboron compounds to imines and nitriles. Several other applications of diboron additions to heteroatom-containing unsaturated organics are also described.

  14. Agrobacterium-mediated transformation of maize (Zea mays) immature embryos.

    PubMed

    Lee, Hyeyoung; Zhang, Zhanyuan J

    2014-01-01

    Agrobacterium tumefaciens-mediated transformation is one of the most efficient and simple gene delivery systems for genetic improvement and biology studies in maize. This system has become more widely used by both public and private laboratories. However, transformation efficiencies vary greatly from laboratory to laboratory for the same genotype. Here, we illustrate our advanced Agrobacterium-mediated transformation method in Hi-II maize using simple binary vectors. The protocol utilizes immature embryos as starting explants and the bar gene as a selectable marker coupled with bialaphos as a selective agent. The protocol offers efficient transformation results with high reproducibility, provided that some experimental conditions are well controlled. This transformation method, with minor modifications, can be also employed to transform certain maize inbreds.

  15. Altered iron homeostasis involvement in arsenite-mediated cell transformation

    PubMed Central

    Wu, Jing; Eckard, Jonathan; Chen, Haobin; Costa, Max; Frenkel, Krystyna; Huang, Xi

    2010-01-01

    Chronic exposure to low doses of arsenite causes transformation of human osteogenic sarcoma (HOS) cells. Although oxidative stress is considered important in arsenite-induced cell transformation, the molecular and cellular mechanisms by which arsenite transforms human cells are still unknown. In the present study, we investigated whether altered iron homeostasis, known to affect cellular oxidative stress, can contribute to the arsenite-mediated cell transformation. Using arsenite-induced HOS cell transformation as a model, it was found that total iron levels are significantly higher in transformed HOS cells in comparison to parental control HOS cells. Under normal iron metabolism conditions, iron homeostasis is tightly controlled by inverse regulation of ferritin and transferrin receptor (TfR) through iron regulatory proteins (IRP). Increased iron levels in arsenite transformed cells should theoretically lead to higher ferritin and lower TfR in these cells than in controls. However, the results showed that both ferritin and TfR are decreased, apparently through two different mechanisms. A lower ferritin level in cytoplasm was due to the decreased mRNA in the arsenite-transformed HOS cells, while the decline in TfR was due to a lowered IRP-binding activity. By challenging cells with iron, it was further established that arsenite-transformed HOS cells are less responsive to iron treatment than control HOS cells, which allows accumulation of iron in the transformed cells, as exemplified by significantly lower ferritin induction. On the other hand, caffeic acid phenethyl ester (CAPE), an antioxidant previously shown to suppress As-mediated cell transformation, prevents As-mediated ferritin depletion. In conclusion, our results suggest that altered iron homeostasis contributes to arsenite-induced oxidative stress and, thus, may be involved in arsenite-mediated cell transformation. PMID:16443159

  16. Amorphous metal distribution transformers: The energy-efficient alternative

    SciTech Connect

    Garrity, T.F.

    1994-12-31

    Amorphous metal distribution transformers have been commercially available for the past 13 years. During that time, they have realized the promise of exceptionally high core efficiency as compared to silicon steel transformer cores. Utility planners today must consider all options available to meet the requirements of load growth. While additional generation capacity will be added, many demand-side initiatives are being undertaken as complementary programs to generation expansion. The efficiency improvement provided by amorphous metal distribution transformers deserves to be among the demand-side options. The key to understanding the positive impact of amorphous metal transformer efficiency is to consider the aggregate contribution those transformers can make towards demand reduction. It is estimated that distribution transformer core losses comprise at least 1% of the utility`s peak demand. Because core losses are continuous, any significant reduction in their magnitude is of great significance to the planner. This paper describes the system-wide economic contributions amorphous metal distribution transformers can make to a utility and suggests evaluation techniques that can be used. As a conservation tool, the amorphous metal transformer contributes to reduced power plant emissions. Calibration of those emissions reductions is also discussed in the paper.

  17. Agrobacterium-mediated transformation of three freshwater microalgal strains.

    PubMed

    Sanitha, Mary; Radha, Sudhakar; Fatima, Anwar Aliya; Devi, Selvaraju Gayathri; Ramya, Mohandass

    2014-01-01

    Microalgal transformation has gained interest in recent years. Agrobacterium-mediated transformation remains as the most efficient method for the development of transgenic plants and microalgae due to its wide host range, inexpensive procedure and transfer of large segments of DNA. In the present study, three different microalgal species were isolated from freshwater environment and identified based on the morphological characteristics and ITS-2 region amplification. Agrobacterium-mediated transformation was successful for the isolates Chlorella sp., Ankistrodesmus sp and Scenedesmus bajacalifornicus. Gene integration and expression was confirmed by PCR amplification of hptII and GUS histochemical assay. A. tumifaciens contamination was checked by amplification of npt II gene (kanamycin resistant) which lies outside the T-border. Based on GUS assay, transformation efficiencies were found to be 12.25% for Chlorella sp. 2.96% for Scenedesmus bajacalifornicus and 3.5% for Ankistrodesmus sp.

  18. TRACE METAL TRANSFORMATION MECHANISMS DURING COAL COMBUSTION

    EPA Science Inventory

    The article reviews mechanisms governing the fate of trace metals during coal combustion and presents new theoretical results that interpret existing data. Emphasis is on predicting the size-segregated speciation of trace metals in pulverized-coal-fired power plant effluents. Thi...

  19. TRACE METAL TRANSFORMATION MECHANISMS DURING COAL COMBUSTION

    EPA Science Inventory

    The article reviews mechanisms governing the fate of trace metals during coal combustion and presents new theoretical results that interpret existing data. Emphasis is on predicting the size-segregated speciation of trace metals in pulverized-coal-fired power plant effluents. Thi...

  20. Methylidene rare-earth-metal complex mediated transformations of C=N, N=N and N-H bonds: new routes to imido rare-earth-metal clusters.

    PubMed

    Hong, Jianquan; Zhang, Lixin; Wang, Kai; Zhang, Yin; Weng, Linhong; Zhou, Xigeng

    2013-06-10

    Three new patterns of reactivity of rare-earth metal methylidene complexes have been established and thus have resulted in access to a wide variety of imido rare-earth metal complexes [L3Ln3(μ2-Me)3(μ3-Me)(μ-NR)] (L = [PhC(NC6H3iPr2-2,6)2](-); R = Ph, Ln = Y (2 a), Lu (2 b); R = 2,6-Me2C6H3, Ln = Y (3 a), Lu (3 b); R = p-ClC6H4, Ln = Y (4 a), Lu (4 b); R = p-MeOC6H4, Ln = Y (5 a), Lu (5 b); R = Me2CHCH2CH2, Ln = Y (6 a), Lu (6 b)) and [{L3Lu3(μ2-Me)3(μ3-Me)}2(μ-NR'N)] (R' = (CH2)6 (7 b), (C6H4)2 (8 b)). Complex 2 b was treated with an excess of CO2 to give the corresponding carboxylate complex [L3Lu3(μ-η(1):η(1)-O2CCH3)3(μ-η(1):η(2)-O2C-CH3)(μ-η(1):η(1):η(2)-O2CNPh)] (9 b) easily. Complex 2 a could undergo the selective μ3-Me abstraction reaction with phenyl acetylene to give the mixed imido/alkynide complex [L3Y3(μ2-Me)3(μ3-η(1):η(1):η(3)-NPh)(μ3-C≡CPh)] (10 a) in high yield. Treatment of 2 with one equivalent of thiophenol gave the selective μ3-methyl-abstracted products [L3Ln3(μ2-Me)3(μ3-η(1):η(1):η(3)-NPh)(μ3-SPh)] (Ln = Y (11 a); Lu (11 b). All new complexes have been characterized by elemental analysis, NMR spectroscopy, and most of the structures confirmed by X-ray diffraction.

  1. Nacnac‐Cobalt‐Mediated P4 Transformations

    PubMed Central

    Spitzer, Fabian; Graßl, Christian; Balázs, Gábor; Mädl, Eric; Keilwerth, Martin; Zolnhofer, Eva M.; Meyer, Karsten

    2017-01-01

    Abstract A comparison of P4 activations mediated by low‐valent β‐diketiminato (L) cobalt complexes is presented. The formal Co0 source [K2(L3Co)2(μ2:η1,η1‐N2)] (1) reacts with P4 to form a mixture of the monoanionic complexes [K(thf)6][(L3Co)2(μ2:η4,η4‐P4)] (2) and [K(thf)6][(L3Co)2(μ2:η3,η3‐P3)] (3). The analogue CoI precursor [L3Co(tol)] (4 a), however, selectively yields the corresponding neutral derivative [(L3Co)2(μ2:η4,η4‐P4)] (5 a). Compound 5 a undergoes thermal P atom loss to form the unprecedented complex [(L3Co)2(μ2:η3,η3‐P3)] (6). The products 2 and 3 can be obtained selectively by an one‐electron reduction of their neutral precursors 5 a and 6, respectively. The electrochemical behaviour of 2, 3, 5 a, and 6 is monitored by cyclic voltammetry and their magnetism is examined by SQUID measurements and the Evans method. The initial CoI‐mediated P4 activation is not influenced by applying the structurally different ligands L1 and L2, which is proven by the formation of the isostructural products [(LCo)2(μ2:η4,η4‐P4)] [L=L3 (5 a), L1 (5 b), L2 (5 c)]. PMID:28032678

  2. Optical characterisation of metallic meshes for space antennas transformable reflectors

    NASA Astrophysics Data System (ADS)

    Mironov, RA; Reznik, SV; Rukavishnikov, RV; Shishulina, VA; Zavaruev, VA

    2016-10-01

    The paper deals with metallic meshes for space antenna transformable reflectors. Optical characteristics of four metallic meshes (15 micron tungsten, 15 micron gold-plated tungsten, 20 micron molybdenum, and 50 micron Ni-coated steel) were investigated with the Nikolet IS50 spectrophotometer. The spectral optical characteristics of these metallic meshes in the range from 0.4 to 2.5 microns are required for thermal design of space antenna reflectors.

  3. Agrobacterium-mediated in planta genetic transformation of sugarcane setts.

    PubMed

    Mayavan, Subramanian; Subramanyam, Kondeti; Jaganath, Balusamy; Sathish, Dorairaj; Manickavasagam, Markandan; Ganapathi, Andy

    2015-10-01

    An efficient, reproducible, and genotype-independent in planta transformation has been developed for sugarcane using setts as explant. Traditional Agrobacterium-mediated genetic transformation and in vitro regeneration of sugarcane is a complex and time-consuming process. Development of an efficient Agrobacterium-mediated transformation protocol, which can produce a large number of transgenic plants in short duration is advantageous. Hence, in the present investigation, we developed a tissue culture-independent in planta genetic transformation system for sugarcane using setts collected from 6-month-old sugarcane plants. The sugarcane setts (nodal cuttings) were infected with three Agrobacterium tumefaciens strains harbouring pCAMBIA 1301-bar plasmid, and the transformants were selected against BASTA(®). Several parameters influencing the in planta transformation such as A. tumefaciens strains, acetosyringone, sonication and exposure to vacuum pressure, have been evaluated. The putatively transformed sugarcane plants were screened by GUS histochemical assay. Sugarcane setts were pricked and sonicated for 6 min and vacuum infiltered for 2 min at 500 mmHg in A. tumefaciens C58C1 suspension containing 100 µM acetosyringone, 0.1 % Silwett L-77 showed the highest transformation efficiency of 29.6 % (with var. Co 62175). The three-stage selection process completely eliminated the chimeric transgenic sugarcane plants. Among the five sugarcane varieties evaluated using the standardized protocol, var. Co 6907 showed the maximum transformation efficiency (32.6 %). The in planta transformation protocol described here is applicable to transfer the economically important genes into different varieties of sugarcane in relatively short time.

  4. Agrobacterium-mediated genetic transformation of Prunus salicina

    USDA-ARS?s Scientific Manuscript database

    We report Agrobacterium tumefaciens-mediated transformation from hypocotyls slices of two Prunus salicina varieties, 'Angeleno' and 'Larry Anne', using a modification of the technique previously described for P. domestica. Regeneration rates on thidiazuron (TDZ) and indole-3-butyric acid (IBA) supp...

  5. Agrobacterium-mediated genetic transformation of Fraxinus americana hypocotyls

    Treesearch

    Kaitlin J. Palla; Paula M. Pijut

    2015-01-01

    An Agrobacterium tumefaciens-mediated genetic transformation system was successfully developed for white ash (Fraxinus americana) using hypocotyls as the initial explants. Hypocotyls isolated from mature embryos germinated on Murashige and Skoog (MS) medium supplemented with 22.2 µM 6-benzyladenine (BA) and 0.5 µM...

  6. Agrobacterium tumefaciens-mediated transformation of Botryosphaeria dothidea.

    PubMed

    Chen, Liang; Wang, Qun; Chen, Hua; Sun, Gengwu; Liu, Huixiang; Wang, Hongkai

    2016-07-01

    Botryosphaeria dothidea is a severe causal agent of die-back and cankers of many woody plants and causes great losses in many regions. The pathogenic mechanism of this pathogen has not been well explored due to lack of mutants and genetic information. In this study, we developed an Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for B. dothidea protoplasts using vector pBHt2 containing the hph gene as a selection marker under the control of trp C promoter. Using this protocol we successfully generated the B. dothidea transformants with efficiency about 23 transformants per 10(5) protoplasts. This is the first report of genetic transformation of B. dothidea via ATMT and this protocol provides an effective tool for B. dothidea genome manipulation, gene identification and functional analysis.

  7. Reversible metal-hydride phase transformation in epitaxial films

    NASA Astrophysics Data System (ADS)

    Roytburd, Alexander L.; Boyerinas, Brad M.; Bruck, Hugh A.

    2015-03-01

    Metal-hydride phase transformations in solids commonly proceed with hysteresis. The extrinsic component of hysteresis is the result of the dissipation of energy of internal stress due to plastic deformation and fracture. It can be mitigated on the nanoscale, where plastic deformation and fracture are suppressed and the transformation proceeds through formation and evolution of coherent phases. However, the phase coherency introduces intrinsic thermodynamic hysteresis, preventing reversible transformation. In this paper, it is shown that thermodynamic hysteresis of coherent metal-hydride transformation can be eliminated in epitaxial film due to substrate constraint. Film-substrate interaction leads to formation of heterophase polydomain nanostructure with variable phase fraction which can change reversibly by varying temperature in a closed system or chemical potential in an open system.

  8. Reversible metal-hydride phase transformation in epitaxial films.

    PubMed

    Roytburd, Alexander L; Boyerinas, Brad M; Bruck, Hugh A

    2015-03-11

    Metal-hydride phase transformations in solids commonly proceed with hysteresis. The extrinsic component of hysteresis is the result of the dissipation of energy of internal stress due to plastic deformation and fracture. It can be mitigated on the nanoscale, where plastic deformation and fracture are suppressed and the transformation proceeds through formation and evolution of coherent phases. However, the phase coherency introduces intrinsic thermodynamic hysteresis, preventing reversible transformation. In this paper, it is shown that thermodynamic hysteresis of coherent metal-hydride transformation can be eliminated in epitaxial film due to substrate constraint. Film-substrate interaction leads to formation of heterophase polydomain nanostructure with variable phase fraction which can change reversibly by varying temperature in a closed system or chemical potential in an open system.

  9. Diverse transformations of liquid metals between different morphologies.

    PubMed

    Sheng, Lei; Zhang, Jie; Liu, Jing

    2014-09-10

    Transformation from a film into a sphere, rapid merging of separate objects, controlled self-rotation, and planar locomotion are the very unusual phenomena observed in liquid metals under application of an electric field to a liquid metal immersed in or sprayed with water. A mechanism for these effects is suggested and potential applications - for example the recovery of liquid metal previously injected into the body for therapeutic purposes - are outlined. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Modeling of Heavy Metal Transformation in Soil Ecosystem

    NASA Astrophysics Data System (ADS)

    Kalinichenko, Kira; Nikovskaya, Galina N.

    2017-04-01

    The intensification of industrial activity leads to an increase in heavy metals pollution of soils. In our opinion, sludge from biological treatment of municipal waste water, stabilized under aerobic-anaerobic conditions (commonly known as biosolid), may be considered as concentrate of natural soil. In their chemical, physical and chemical and biological properties these systems are similar gel-like nanocomposites. These contain microorganisms, humic substances, clay, clusters of nanoparticles of heavy metal compounds, and so on involved into heteropolysaccharides matrix. It is known that microorganisms play an important role in the transformation of different nature substances in soil and its health maintenance. The regularities of transformation of heavy metal compounds in soil ecosystem were studied at the model of biosolid. At biosolid swelling its structure changing (gel-sol transition, weakening of coagulation contacts between metal containing nanoparticles, microbial cells and metabolites, loosening and even destroying of the nanocomposite structure) can occur [1, 2]. The promotion of the sludge heterotrophic microbial activities leads to solubilization of heavy metal compounds in the system. The microbiological process can be realized in alcaligeneous or acidogeneous regimes in dependence on the type of carbon source and followed by the synthesis of metabolites with the properties of flocculants and heavy metals extragents [3]. In this case the heavy metals solubilization (bioleaching) in the form of nanoparticles of hydroxycarbonate complexes or water soluble complexes with oxycarbonic acids is observed. Under the action of biosolid microorganisms the heavy metals-oxycarbonic acids complexes can be transformed (catabolised) into nano-sizing heavy metals- hydroxycarbonates complexes. These ecologically friendly complexes and microbial heteropolysaccharides are able to interact with soil colloids, stay in the top soil profile, and improve soil structure due

  11. TEADs mediate nuclear retention of TAZ to promote oncogenic transformation.

    PubMed

    Chan, Siew Wee; Lim, Chun Jye; Loo, Li Shen; Chong, Yaan Fun; Huang, Caixia; Hong, Wanjin

    2009-05-22

    The transcriptional coactivators YAP and TAZ are downstream targets inhibited by the Hippo tumor suppressor pathway. The expression level of TAZ is recently shown to be elevated in invasive breast cancer cells and some primary breast cancers. TAZ is important for breast cancer cell migration, invasion, and tumorigenesis, but the underlying mechanism is not defined. In this study, we show that TAZ interacts with TEAD transcriptional factors. Knockdown of TEADs suppresses TAZ-mediated oncogenic transformation of MCF10A cells. Uncoupling TAZ from Hippo regulation by S89A mutation enhances its transforming ability. Several residues located in the N-terminal region of TAZ are identified to be important for interaction with TEADs, and these same residues are equally important for TAZ to transform MCF10A cells. Mechanistically, TAZ mutants defective in interaction with TEADs fail to accumulate in the nucleus. Live cell imaging of enhanced green fluorescent protein-TAZ and its mutant defective in TEAD interaction suggests that TEAD interaction mediates nuclear retention. These results reveal a novel mechanism for TEADs to regulate nuclear retention and thus the transforming ability of TAZ.

  12. Progress of cereal transformation technology mediated by Agrobacterium tumefaciens

    PubMed Central

    Hiei, Yukoh; Ishida, Yuji; Komari, Toshihiko

    2014-01-01

    Monocotyledonous plants were believed to be not transformable by the soil bacterium Agrobacterium tumefaciens until two decades ago, although convenient protocols for infection of leaf disks and subsequent regeneration of transgenic plants had been well established in a number of dicotyledonous species by then. This belief was reinforced by the fact that monocotyledons are mostly outside the host range of crown gall disease caused by the bacterium and by the failures in trials in monocotyledons to mimic the transformation protocols for dicotyledons. However, a key reason for the failure could have been the lack of active cell divisions at the wound sites in monocotyledons. The complexity and narrow optimal windows of critical factors, such as genotypes of plants, conditions of the plants from which explants are prepared, tissue culture methods and culture media, pre-treatments of explants, strains of A. tumefaciens, inducers of virulence genes, transformation vectors, selection marker genes and selective agents, kept technical hurdles high. Eventually it was demonstrated that rice and maize could be transformed by co-cultivating cells of callus cultures or immature embryos, which are actively dividing or about to divide, with A. tumefaciens. Subsequently, these initial difficulties were resolved one by one by many research groups, and the major cereals are now transformed quite efficiently. As many as 15 independent transgenic events may be regenerated from a single piece of immature embryo of rice. Maize transformation protocols are well established, and almost all transgenic events deregulated for commercialization after 2003 were generated by Agrobacterium-mediated transformation. Wheat, barley, and sorghum are also among those plants that can be efficiently transformed by A. tumefaciens. PMID:25426132

  13. Progress of cereal transformation technology mediated by Agrobacterium tumefaciens.

    PubMed

    Hiei, Yukoh; Ishida, Yuji; Komari, Toshihiko

    2014-01-01

    Monocotyledonous plants were believed to be not transformable by the soil bacterium Agrobacterium tumefaciens until two decades ago, although convenient protocols for infection of leaf disks and subsequent regeneration of transgenic plants had been well established in a number of dicotyledonous species by then. This belief was reinforced by the fact that monocotyledons are mostly outside the host range of crown gall disease caused by the bacterium and by the failures in trials in monocotyledons to mimic the transformation protocols for dicotyledons. However, a key reason for the failure could have been the lack of active cell divisions at the wound sites in monocotyledons. The complexity and narrow optimal windows of critical factors, such as genotypes of plants, conditions of the plants from which explants are prepared, tissue culture methods and culture media, pre-treatments of explants, strains of A. tumefaciens, inducers of virulence genes, transformation vectors, selection marker genes and selective agents, kept technical hurdles high. Eventually it was demonstrated that rice and maize could be transformed by co-cultivating cells of callus cultures or immature embryos, which are actively dividing or about to divide, with A. tumefaciens. Subsequently, these initial difficulties were resolved one by one by many research groups, and the major cereals are now transformed quite efficiently. As many as 15 independent transgenic events may be regenerated from a single piece of immature embryo of rice. Maize transformation protocols are well established, and almost all transgenic events deregulated for commercialization after 2003 were generated by Agrobacterium-mediated transformation. Wheat, barley, and sorghum are also among those plants that can be efficiently transformed by A. tumefaciens.

  14. Microbial-mediated method for metal oxide nanoparticle formation

    DOEpatents

    Rondinone, Adam J.; Moon, Ji Won; Love, Lonnie J.; Yeary, Lucas W.; Phelps, Tommy J.

    2015-09-08

    The invention is directed to a method for producing metal oxide nanoparticles, the method comprising: (i) subjecting a combination of reaction components to conditions conducive to microbial-mediated formation of metal oxide nanoparticles, wherein said combination of reaction components comprise: metal-reducing microbes, a culture medium suitable for sustaining said metal-reducing microbes, an effective concentration of one or more surfactants, a reducible metal oxide component containing one or more reducible metal species, and one or more electron donors that provide donatable electrons to said metal-reducing microbes during consumption of the electron donor by said metal-reducing microbes; and (ii) isolating said metal oxide nanoparticles, which contain a reduced form of said reducible metal oxide component. The invention is also directed to metal oxide nanoparticle compositions produced by the inventive method.

  15. Structure Fluctuation Model of Melting and Polymorphic Transformations in Metals

    NASA Astrophysics Data System (ADS)

    Filippov, E. S.

    2017-09-01

    Relationships of volumes during thermal expansion of metals are analyzed. It is demonstrated that metals with BCC structure undergo two-step structural change at the melting point: at the beginning the BCC structure is transformed into FCC structure and then clusters are formed with K = 12 and statistical packing of atoms of the liquid phase. Before melting, hexagonal layer-by-layer (6 + 6) packed Cd and Zn change their structure from K = 6 to K = 8. Metals with FCC structure do not change the number of neighbors before melting, forming clusters with K = 12. It is shown that conditions of pre-melting and polymorphic FCC (HDP) → BCC transformations are reached for the critical volume of thermal expansion caused by leveling of fluctuations of atom and electron densities.

  16. Optimization of Agrobacterium-Mediated Transformation in Soybean

    PubMed Central

    Li, Shuxuan; Cong, Yahui; Liu, Yaping; Wang, Tingting; Shuai, Qin; Chen, Nana; Gai, Junyi; Li, Yan

    2017-01-01

    High transformation efficiency is a prerequisite for study of gene function and molecular breeding. Agrobacterium tumefaciens-mediated transformation is a preferred method in many plants. However, the transformation efficiency in soybean is still low. The objective of this study is to optimize Agrobacterium-mediated transformation in soybean by improving the infection efficiency of Agrobacterium and regeneration efficiency of explants. Firstly, four factors affecting Agrobacterium infection efficiency were investigated by estimation of the rate of GUS transient expression in soybean cotyledonary explants, including Agrobacterium concentrations, soybean explants, Agrobacterium suspension medium, and co-cultivation time. The results showed that an infection efficiency of over 96% was achieved by collecting the Agrobacterium at a concentration of OD650 = 0.6, then using an Agrobacterium suspension medium containing 154.2 mg/L dithiothreitol to infect the half-seed cotyledonary explants (from mature seeds imbibed for 1 day), and co-cultured them for 5 days. The Agrobacterium infection efficiencies for soybean varieties Jack Purple and Tianlong 1 were higher than the other six varieties. Secondly, the rates of shoot elongation were compared among six different concentration combinations of gibberellic acid (GA3) and indole-3-acetic acid (IAA). The shoot elongation rate of 34 and 26% was achieved when using the combination of 1.0 mg/L GA3 and 0.1 mg/L IAA for Jack Purple and Tianlong 1, respectively. This rate was higher than the other five concentration combinations of GA3 and IAA, with an 18 and 11% increase over the original laboratory protocol (a combination of 0.5 mg/L GA3 and 0.1 mg/L IAA), respectively. The transformation efficiency was 7 and 10% for Jack Purple and Tianlong 1 at this optimized hormone concentration combination, respectively, which was 2 and 6% higher than the original protocol, respectively. Finally, GUS histochemical staining, PCR, herbicide

  17. Highly efficient transformation system for Malassezia furfur and Malassezia pachydermatis using Agrobacterium tumefaciens-mediated transformation.

    PubMed

    Celis, A M; Vos, A M; Triana, S; Medina, C A; Escobar, N; Restrepo, S; Wösten, H A B; de Cock, H

    2017-03-01

    Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M. furfur and M. pachydermatis by combining the transformation protocols of Agaricus bisporus and Cryptococcus neoformans. Optimal temperature and co-cultivation time for transformation were 5 and 7days at 19°C and 24°C, respectively. Transformation efficiency was 0.75-1.5% for M. furfur and 0.6-7.5% for M. pachydermatis. Integration of the hpt resistance cassette and gfp was verified using PCR and fluorescence microscopy, respectively. The T-DNA was mitotically stable in approximately 80% of the transformants after 10 times sub-culturing in the absence of hygromycin. Improving transformation protocols contribute to study the biology and pathophysiology of Malassezia.

  18. Frugivore-Mediated Selection in A Habitat Transformation Scenario

    PubMed Central

    Fontúrbel, Francisco E.; Medel, Rodrigo

    2017-01-01

    Plant-animal interactions are strong drivers of phenotypic evolution. However, the extent to which anthropogenic habitat transformation creates new selective scenarios for plant-animal interactions is a little explored subject. We examined the effects of native forest replacement by exotic Eucalyptus trees on the frugivore-mediated phenotypic selection coefficients imposed by the relict marsupial Dromiciops gliroides upon traits involved in frugivore attraction and germination success of the mistletoe Tristerix corymbosus (Loranthaceae). We found significant gradients for seed weight and sugar content along the native - transformed habitat gradient. While selection for larger seed weight was more relevant in native habitats, fruits with intermediate sugar content were promoted in transformed habitats. The spatial habitat structure and microclimate features such as the degree of sunlight received influenced the natural selection processes, as they correlated with the phenotypic traits analysed. The response of this plant-frugivore interaction to human disturbance seemed to be context-dependent, in which extremely transformed habitats would offer new opportunities for natural selection on dispersal-related traits. Even in recent transformation events like this, human disturbance acts as a strong contemporary evolution driver. PMID:28349942

  19. Frugivore-Mediated Selection in A Habitat Transformation Scenario.

    PubMed

    Fontúrbel, Francisco E; Medel, Rodrigo

    2017-03-28

    Plant-animal interactions are strong drivers of phenotypic evolution. However, the extent to which anthropogenic habitat transformation creates new selective scenarios for plant-animal interactions is a little explored subject. We examined the effects of native forest replacement by exotic Eucalyptus trees on the frugivore-mediated phenotypic selection coefficients imposed by the relict marsupial Dromiciops gliroides upon traits involved in frugivore attraction and germination success of the mistletoe Tristerix corymbosus (Loranthaceae). We found significant gradients for seed weight and sugar content along the native - transformed habitat gradient. While selection for larger seed weight was more relevant in native habitats, fruits with intermediate sugar content were promoted in transformed habitats. The spatial habitat structure and microclimate features such as the degree of sunlight received influenced the natural selection processes, as they correlated with the phenotypic traits analysed. The response of this plant-frugivore interaction to human disturbance seemed to be context-dependent, in which extremely transformed habitats would offer new opportunities for natural selection on dispersal-related traits. Even in recent transformation events like this, human disturbance acts as a strong contemporary evolution driver.

  20. Transformation of heavy metal speciation during sludge drying: mechanistic insights.

    PubMed

    Weng, Huan-Xin; Ma, Xue-Wen; Fu, Feng-Xia; Zhang, Jin-Jun; Liu, Zan; Tian, Li-Xun; Liu, Chongxuan

    2014-01-30

    Speciation can fundamentally affect on the stability and toxicity of heavy metals in sludge from wastewater treatment plants. This research investigated the speciation of heavy metals in sludge from both municipal and industrial sources, and metal speciation change as a result of drying process to reduce sludge volume. The changes in sludge properties including sludge moisture content, temperature, density, and electrical conductivity were also monitored to provide insights into the mechanisms causing the change in heavy metal speciation. The results show that the drying process generally stabilized Cr, Cu, Cd, and Pb in sludge by transforming acid-soluble, reducible, and oxidizable species into structurally stable forms. Such transformation and stabilization occurred regardless of the sludge source and type, and were primarily caused by the changes in sludge properties associated with decomposition of organic matter and sulfide. The results enhanced our understanding of the geochemical behavior of heavy metals in municipal sludge, and are useful for designing a treatment system for environment-friendly disposal of sludge.

  1. Transformation of heavy metal speciation during sludge drying: mechanistic insights

    SciTech Connect

    Weng, Huanxin; Ma, Xue-Wen; Fu, Feng-Xia; Zhang, Jin-Jun; Liu, Zan; Tian, Li-Xun; Liu, Chongxuan

    2014-01-30

    Speciation can fundamentally affect on the stability and toxicity of heavy metals in sludge from wastewater treatment plants. This research investigated the speciation of heavy metals in sludge from both municipal and industrial sources, and metal speciation change as a result of drying process to reduce sludge volume. The changes in sludge properties including sludge moisture content, temperature, density, and electrical conductivity were also monitored to provide insights into the mechanisms causing the change in heavy metal speciation. The results show that the drying process generally stabilized the Cr, Cu, Cd and Pb in sludge by transforming acid-soluble, reducible and oxidizable species into structurally stable forms. Such transformation and stabilization occurred regardless of the sludge source and type, and were primarily caused by the changes in sludge properties associated with decomposition of organic matter and sulfide. The results enhanced our understanding of the geochemical behavior of heavy metals in municipal sludge, and are useful for designing a treatment system for environment-friendly disposal of sludge.

  2. [Transformation of Didymella bryoniae mediated by Agrobacterium tumefaciens].

    PubMed

    Ren, Haiying; Fang, Li; Li, Gang; Ru, Shuijiang; Wang, Hanrong

    2010-06-01

    Gummy stem blight, a plant disease caused by Didymella bryoniae, is one of the major diseases in melon. The disease can seriously reduce melon yield and quality. However, little information is available on the genetics and functional genomics of the fungal pathogen. In this study, we developed an Agrobacterium-mediated transformation system for D. bryoniae by using a universal pathogenic isolate DB11 and the Agrobacterium tumefaciens strain C58C1 carrying plasmid pBIG2RHPH2 harboring the hygromycin B phosphotransferase gene (hph). Total 45 transformants could be obtained per 1 x 10(5) spores when 1 x 10(6) spores per milliliter of D. bryoniae spore suspension were cocultivated with Agrobacterium cells at OD600 = 0.15 for 48 h in the presence of induction medium (pH 5.2) containing acetosyringone at 200 microg/mL and selection medium contained 100 microg/mL of hygromycin B and 200 microg/mL of cefotaxime sodium, ampicillin and tetracycline, respectively. The transformants were stable when grown on PDA medium without hygromycin B for five times and were verified by PCR amplification with the hph primers and by Southern blot analysis with the hph probe. The transformation system will be useful for further studies of functional genes in D. bryoniae.

  3. Folding/Unfolding Properties of Metal Foils in Transformable Structure

    NASA Astrophysics Data System (ADS)

    Daming, Nie; Zhen, Lu; Kaifeng, Zhang

    2016-11-01

    Transformable structures are widely applied in the aerospace, temporary facilities, etc. Compared to the structures made of polyester materials, the metal foil ones occupy many special advantages while have been rarely investigated. In this study, a series of transformable structures made of four different metal materials, 6065 Al, copper, TA1 and SUS 304 stainless steel, with thickness of 0.1 mm were prepared. Moreover, the folding (i.e., compressing the structure to the lowest height with external force) and unfolding (i.e., extending the structure to the largest height with external force) behaviors of these structures were exhibited and explained by experiments. Besides, the differences and corresponding mechanisms of various materials on the folding/unfolding properties of the structures were examined and discussed.

  4. Folding/Unfolding Properties of Metal Foils in Transformable Structure

    NASA Astrophysics Data System (ADS)

    Daming, Nie; Zhen, Lu; Kaifeng, Zhang

    2017-01-01

    Transformable structures are widely applied in the aerospace, temporary facilities, etc. Compared to the structures made of polyester materials, the metal foil ones occupy many special advantages while have been rarely investigated. In this study, a series of transformable structures made of four different metal materials, 6065 Al, copper, TA1 and SUS 304 stainless steel, with thickness of 0.1 mm were prepared. Moreover, the folding (i.e., compressing the structure to the lowest height with external force) and unfolding (i.e., extending the structure to the largest height with external force) behaviors of these structures were exhibited and explained by experiments. Besides, the differences and corresponding mechanisms of various materials on the folding/unfolding properties of the structures were examined and discussed.

  5. Reflectance Changes during Shock-induced Phase Transformations in Metals

    SciTech Connect

    Stevens, G. D.; Turley, W. D.; Veeser, L. R.; Jensen, B. J.; Rigg, P. A.

    2010-06-01

    In performing shock wave experiments to study the characteristics of metals at high pressures, wave profiles (i.e., velocity measurements of the surface of the sample) are an established and useful way to study phase transformations. For example, a sudden change in the velocity or its slope can occur when the phase transformation induces a large volume change leading to a change in particle velocity. Allowing the shock to release into a transparent window that is in contact with the sample surface allows the study of conditions away from the shock Hugoniot. However, in cases where the wave profile is not definitive an additional phase-transformation diagnostic would be useful. Changes in the electronic structure of the atoms in the crystal offer opportunities to develop new phase-change diagnostics. We have studied optical reflectance changes for several phase transformations to see whether reflectance changes might be a generally applicable phase-transformation diagnostic. Shocks were produced by direct contact with explosives or with impacts from guns. Optical wavelengths for the reflectance measurements ranged from 355 to 700 nm. We studied samples of tin, iron, gallium, and cerium as each passed through a phase transformation during shock loading and, if observable, a reversion upon unloading. For solid-solid phase changes in tin and iron we saw small changes in the surface scattering characteristics, perhaps from voids or rough areas frozen into the surface of the sample as it transformed to a new crystal structure. For melt in gallium and cerium we saw changes in the wavelength dependence of the reflectance, and we surmise that these changes may result from changes in the crystal electronic structure. It appears that reflectance measurements can be a significant part of a larger suite of diagnostics to search for difficult-to-detect phase transformations.

  6. Exploiting Semiconductor to Metallic Phase Transformation in Layered Transition Metal Dichalcogenides for Ohmic contact Contacts

    NASA Astrophysics Data System (ADS)

    Kappera, Rajesh; Voiry, Damien; Jen, Wesley; Yalcin, Sibel Ebru; Gupta, Gautam; Mohite, Aditya; Chhowalla, Manish; Material Science department, Rutgers University, Piscataway, NJ, 08854, USA Team; CenterIntegrated Nanotechnologies, Los Alamos National Laboratory, Los Alamos, NM, 87544, US Team

    2014-03-01

    Achieving ohmic contacts to transition metal dichalcogenides (MoS2, WS2, WSe2 and MoSe2) has been a challenge for researchers owing to the formation of a large Schottky barrier between metal and semiconductor. This results in low on-currents, mobilities and sub-threshold swings in the devices made with these materials. Here we report a universal strategy using chemical approach to reversibly transform the semiconducting phase (2H) to metallic phase (1T). Taking advantage of the metallic phase, we have fabricated hybrid transistors, which have 1T phase contacts and semiconducting 2H phase of the material as the channel. The metallic phase dramatically reduces the Schottky barrier between the metal and the semiconductor thereby mitigating the high contact resistance issues. This strategy should be applicable to several other applications such as catalysis, supercapacitors and batteries. Detailed synthesis, structural, electrical and optical characterization will be described.

  7. Oncogene-mediated transformation of fetal rat colon in vitro.

    PubMed

    Pories, S; Jaros, K; Steele, G; Pauley, A; Summerhayes, I C

    1992-05-01

    Short-term maintenance of fetal rat colonic tissue in vitro has been demonstrated using a collagen matrix organ culture system. The introduction of single (v-myc, v-rasH, v-src) oncogenes or combinations of oncogenes (v-myc/rasH, v-myc/src) into normal colon mucosal elements was established using retroviral vectors, resulting in enhanced proliferation and migration of epithelial cells from the lumen of tissue implants. Expression of a single oncogene in normal colon epithelium did not result in the establishment of cell lines. In contrast, expression of cooperating oncogenic elements resulted in cell lines in greater than 80% of experiments, revealing different morphological characteristics dependent upon the oncogene combination used. Confirmation of the expression of viral transcripts was determined using Northern blot analysis and viral oncoprotein expression using Western blot analysis (p21) and an immunoprecipitation kinase assay (src). Expression of keratin filaments was lost following passaging of cell lines but could be induced by the myc/ras transformants by growth on Rat-1 feeder layers. This induction phenomenon was not observed with myc/src lines, and although these expressed high levels of sucrase isomaltase the epithelial origin of these cells is unclear. Karyotypic analysis performed on three myc/ras-transformed cell lines revealed a normal chromosome complement associated with transformation. In this report we describe a novel in vitro transformation system for normal rat colonic epithelium mediated by the introduction of oncogene elements using different retroviral vector constructs. The potential to generate cell lines representing different stages of neoplastic progression using relevant genetic components presents significant advantages for the study of cellular and molecular interactions underlying colon neoplastic progression.

  8. Biomolecular Mechanisms Controlling Metal and Radionuclide Transformations in Anaeromyxobacter dehalogenans

    SciTech Connect

    Beliaev, Alexander S.; Fredrickson, James K.; Loeffler, Frank E.; Sanford, Robert A.

    2006-06-01

    Microbiological reduction and immobilization of U(VI) and Tc(VII) has been proposed as a strategy for remediating radionuclide-contaminated environments. Numerous studies focusing on the reduction kinetics and speciation of these metals have been carried out using contaminated sediment samples, microbial consortia, and pure bacterial cultures. While previous work with model organisms has increased the general understanding of radionuclide transformation processes, fundamental questions regarding radionuclide reduction mechanisms by indigenous microorganisms are poorly understood, especially under the commonly encountered scenario where multiple electron acceptors are present. Therefore, the overall goal of the proposed research is to elucidate the molecular mechanisms of radionuclide biotransformation by Anaeromyxobacter dehalogenans, a predominant member of indigenous microorganism commonly found in contaminated subsurface environments, and to assess the effects of relevant environmental factors affecting these transformation reactions.

  9. Agrobacterium-mediated transformation of bottle gourd (Lagenaria siceraria Standl.).

    PubMed

    Han, J-S; Kim, C K; Park, S H; Hirschi, K D; Mok, I- G

    2005-03-01

    We describe a procedure for producing transgenic bottle gourd plants by inoculating cotyledon explants with Agrobacterium tumefaciens strain AGL1 that carries the binary vector pCAMBIA3301 containing a glufosinate ammonium-resistance (bar) gene and the beta-D-glucuronidase (GUS) reporter gene. The most effective bacterial infection was observed when cotyledon explants of 4-day-old seedlings were co-cultivated with Agrobacterium for 6-8 days on co-cultivation medium supplemented with 0.1-0.001 mg/l L-alpha-(2-aminoethoxyvinyl) glycine (AVG). The putatively transformed shoots directly emerged at the proximal end of cotyledon explants after 2-3 weeks of culturing on selection medium containing 2 mg/l DL-phosphinothricin. These shoots were rooted after 3 weeks of culturing on half-strength MS medium containing 0.1 mg/l indole acetic acid and 1 mg/l DL-phosphinothricin. Transgenic plants were obtained at frequencies of 1.9%. Stable integration and transmission of the transgenes in T1 generation plants were confirmed by a histochemical GUS assay, polymerase chain reaction and Southern blot analyses. Genetic segregation analysis of T1 progenies showed that transgenes were inherited in a Mendelian fashion. To our knowledge, this study is the first to show Agrobacterium-mediated transformation in bottle gourd.

  10. Efficient method for Agrobacterium mediated transformation of Artemisia annua L.

    PubMed

    Alam, Pravej; Mohammad, Anis; Ahmad, M M; Khan, Mather Ali; Nadeem, Mohd; Khan, Riyazuddeen; Akmal, Mohd; Ahlawat, Seema; Abdin, M Z

    2014-01-01

    Artemisinin, a potent antimalarial natural products isolated from aerial parts of Artemisia annua L. Many patents have been reported that the demand for artemisinin is exponentially increasing year after year due to increased incidences of drug resistant malaria throughout the world. Leaf explants were used frequently as target tissue to generate transgenic of Artemisia. annua L. However, obtaining a large number of transgenic lines through out the year is a laborious and delicate process. To circumvent this, we have developed a highly efficient leaf explant based Agrobacterium mediated transformation of A. annua L. plant. The gus gene was used as screenable marker to assess and optimize the performance of T-DNA delivery. The age of explant, kind of bacterial inoculation, suspension duration, infection times and co-culture conditions were optimized. The co-culture was carried out with Agrobacterium tumefaciens strain EHA105 under desiccation condition in the dark at 25-28 0C for 2-4 days. Complete analysis of transgene insertion demonstrated that the optimized method of transformation from leaf explants of A. annua L. was efficient and highly reproducible.

  11. Aboveground insect infestation attenuates belowground Agrobacterium-mediated genetic transformation.

    PubMed

    Song, Geun Cheol; Lee, Soohyun; Hong, Jaehwa; Choi, Hye Kyung; Hong, Gun Hyong; Bae, Dong-Won; Mysore, Kirankumar S; Park, Yong-Soon; Ryu, Choong-Min

    2015-07-01

    Agrobacterium tumefaciens causes crown gall disease. Although Agrobacterium can be popularly used for genetic engineering, the influence of aboveground insect infestation on Agrobacterium induced gall formation has not been investigated. Nicotiana benthamiana leaves were exposed to a sucking insect (whitefly) infestation and benzothiadiazole (BTH) for 7 d, and these exposed plants were inoculated with a tumorigenic Agrobacterium strain. We evaluated, both in planta and in vitro, how whitefly infestation affects crown gall disease. Whitefly-infested plants exhibited at least a two-fold reduction in gall formation on both stem and crown root. Silencing of isochorismate synthase 1 (ICS1), required for salicylic acid (SA) synthesis, compromised gall formation indicating an involvement of SA in whitefly-derived plant defence against Agrobacterium. Endogenous SA content was augmented in whitefly-infested plants upon Agrobacterium inoculation. In addition, SA concentration was three times higher in root exudates from whitefly-infested plants. As a consequence, Agrobacterium-mediated transformation of roots of whitefly-infested plants was clearly inhibited when compared to control plants. These results suggest that aboveground whitefly infestation elicits systemic defence responses throughout the plant. Our findings provide new insights into insect-mediated leaf-root intra-communication and a framework to understand interactions between three organisms: whitefly, N. benthamiana and Agrobacterium. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

  12. Effect of humic acid on transformation of soil heavy metals

    NASA Astrophysics Data System (ADS)

    Wu, Shengzhe; Li, Rui; Peng, Shuyang; Liu, Qiuyong; Zhu, Xi

    2017-06-01

    The pattern of transformation among different fractions of heavy metals in soil draws great attention. This article investigated the effect of humic acid on soil heavy metal under different pH and temperature. The results showed that with the increasing of the concentration of humic acid, the concentration of available Cu, Pb decreased greatly, though the decrease of available Cd was light. Also, pH of soil had certain impact on the concentration of available Cu, Pb, Cd while the influence of environmental temperature was minor. The removal efficiency of contaminated soil was 57.283% (Cu), 2.645% (Cd) and 15.485% (Pb) while it could reach more than 98% in contaminated solution.

  13. Transformation of metal-organic frameworks for molecular sieving membranes

    PubMed Central

    Li, Wanbin; Zhang, Yufan; Zhang, Congyang; Meng, Qin; Xu, Zehai; Su, Pengcheng; Li, Qingbiao; Shen, Chong; Fan, Zheng; Qin, Lei; Zhang, Guoliang

    2016-01-01

    The development of simple, versatile strategies for the synthesis of metal-organic framework (MOF)-derived membranes are of increasing scientific interest, but challenges exist in understanding suitable fabrication mechanisms. Here we report a route for the complete transformation of a series of MOF membranes and particles, based on multivalent cation substitution. Through our approach, the effective pore size can be reduced through the immobilization of metal salt residues in the cavities, and appropriate MOF crystal facets can be exposed, to achieve competitive molecular sieving capabilities. The method can also be used more generally for the synthesis of a variety of MOF membranes and particles. Importantly, we design and synthesize promising MOF membranes candidates that are hard to achieve through conventional methods. For example, our CuBTC/MIL-100 membrane exhibits 89, 171, 241 and 336 times higher H2 permeance than that of CO2, O2, N2 and CH4, respectively. PMID:27090597

  14. Transformation of metal-organic frameworks for molecular sieving membranes

    NASA Astrophysics Data System (ADS)

    Li, Wanbin; Zhang, Yufan; Zhang, Congyang; Meng, Qin; Xu, Zehai; Su, Pengcheng; Li, Qingbiao; Shen, Chong; Fan, Zheng; Qin, Lei; Zhang, Guoliang

    2016-04-01

    The development of simple, versatile strategies for the synthesis of metal-organic framework (MOF)-derived membranes are of increasing scientific interest, but challenges exist in understanding suitable fabrication mechanisms. Here we report a route for the complete transformation of a series of MOF membranes and particles, based on multivalent cation substitution. Through our approach, the effective pore size can be reduced through the immobilization of metal salt residues in the cavities, and appropriate MOF crystal facets can be exposed, to achieve competitive molecular sieving capabilities. The method can also be used more generally for the synthesis of a variety of MOF membranes and particles. Importantly, we design and synthesize promising MOF membranes candidates that are hard to achieve through conventional methods. For example, our CuBTC/MIL-100 membrane exhibits 89, 171, 241 and 336 times higher H2 permeance than that of CO2, O2, N2 and CH4, respectively.

  15. Metal-Support Cooperative Catalysts for Environmentally Benign Molecular Transformations.

    PubMed

    Kaneda, Kiyotomi; Mitsudome, Takato

    2017-01-01

    Metal-support cooperative catalysts have been developed for sustainable and environmentally benign molecular transformations. The active metal centers and supports in these catalysts could cooperatively activate substrates, resulting in high catalytic performance for liquid-phase reactions under mild conditions. These catalysts involved hydrotalcite-supported gold and silver nanoparticles with high catalytic activity for organic reactions such as aerobic oxidation, oxidative carbonylation, and chemoselective reduction of epoxides to alkenes and nitrostyrenes to aminostyrenes using alcohols and CO/H2 O as reducing reagents. This high catalytic performance was due to cooperative catalysis between the metal nanoparticles and basic sites of the hydrotalcite support. To increase the metal-support cooperative effect, core-shell nanostructured catalysts consisting of gold or silver nanoparticles in the core and ceria supports in the shell were designed. These core-shell nanocomposite catalysts were effective for the chemoselective hydrogenation of nitrostyrenes to aminostyrenes, unsaturated aldehydes to allyl alcohols, and alkynes to alkenes using H2 as a clean reductant. In addition, these solid catalysts could be recovered easily from the reaction mixture by simple filtration, and were reusable with high catalytic activity. © 2017 The Chemical Society of Japan & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Metal Complexes for DNA-Mediated Charge Transport

    PubMed Central

    Barton, Jacqueline K.; Olmon, Eric D.; Sontz, Pamela A.

    2010-01-01

    In all organisms, oxidation threatens the integrity of the genome. DNA-mediated charge transport (CT) may play an important role in the generation and repair of this oxidative damage. In studies involving long-range CT from intercalating Ru and Rh complexes to 5′-GG-3′ sites, we have examined the efficiency of CT as a function of distance, temperature, and the electronic coupling of metal oxidants bound to the base stack. Most striking is the shallow distance dependence and the sensitivity of DNA CT to how the metal complexes are stacked in the helix. Experiments with cyclopropylamine-modified bases have revealed that charge occupation occurs at all sites along the bridge. Using Ir complexes, we have seen that the process of DNA-mediated reduction is very similar to that of DNA-mediated oxidation. Studies involving metalloproteins have, furthermore, shown that their redox activity is DNA-dependent and can be DNA-mediated. Long range DNA-mediated CT can facilitate the oxidation of DNA-bound base excision repair proteins to initiate a redox-active search for DNA lesions. DNA CT can also activate the transcription factor SoxR, triggering a cellular response to oxidative stress. Indeed, these studies show that within the cell, redox-active proteins may utilize the same chemistry as that of synthetic metal complexes in vitro, and these proteins may harness DNA-mediated CT to reduce damage to the genome and regulate cellular processes. PMID:21643528

  17. From Nanowires to Biofilms: An Exploration of Novel Mechanisms of Uranium Transformation Mediated by Geobacter Bacteria

    SciTech Connect

    REGUERA, GEMMA

    2014-01-16

    One promising strategy for the in situ bioremediation of radioactive groundwater contaminants that has been identified by the SBR Program is to stimulate the activity of dissimilatory metal-reducing microorganisms to reductively precipitate uranium and other soluble toxic metals. The reduction of U(VI) and other soluble contaminants by Geobacteraceae is directly dependent on the reduction of Fe(III) oxides, their natural electron acceptor, a process that requires the expression of Geobacter’s conductive pili (pilus nanowires). Expression of conductive pili by Geobacter cells leads to biofilm development on surfaces and to the formation of suspended biogranules, which may be physiological closer to biofilms than to planktonic cells. Biofilm development is often assumed in the subsurface, particularly at the matrix-well screen interface, but evidence of biofilms in the bulk aquifer matrix is scarce. Our preliminary results suggest, however, that biofilms develop in the subsurface and contribute to uranium transformations via sorption and reductive mechanisms. In this project we elucidated the mechanism(s) for uranium immobilization mediated by Geobacter biofilms and identified molecular markers to investigate if biofilm development is happening in the contaminated subsurface. The results provided novel insights needed in order to understand the metabolic potential and physiology of microorganisms with a known role in contaminant transformation in situ, thus having a significant positive impact in the SBR Program and providing novel concept to monitor, model, and predict biological behavior during in situ treatments.

  18. Transition-metal-mediated thiosulfinate ester synthesis

    SciTech Connect

    Raseta, M.E.; Cawood, S.A.; Welker, M.E. ); Rheingold, A.L. )

    1989-10-11

    Unlike sulfur dioxide (SO{sub 2}), the coordination and organic reaction chemistry of disulfur monoxide (S{sub 2}O) has received little attention. A few Diels-Alder reactions of simple dienes with S{sub 2}O have been reported. However, there were no reports of direct S{sub 2}O complex synthesis prior to our initial work. The authors recently published a synthesis of 4,5-diphenyl-3,6-dihydro-1,2-dithiin 1-oxide (2) which liberates S{sub 2}O via a transition-metal-assisted retro-Diels-Alder reaction. Here we report further on the unusual reactivity of 2 and its utilization in the synthesis of cyclic thiosulfinate esters.

  19. Biosurfactant Mediated Biosynthesis of Selected Metallic Nanoparticles

    PubMed Central

    Płaza, Grażyna A.; Chojniak, Joanna; Banat, Ibrahim M.

    2014-01-01

    Developing a reliable experimental protocol for the synthesis of nanomaterials is one of the challenging topics in current nanotechnology particularly in the context of the recent drive to promote green technologies in their synthesis. The increasing need to develop clean, nontoxic and environmentally safe production processes for nanoparticles to reduce environmental impact, minimize waste and increase energy efficiency has become essential in this field. Consequently, recent studies on the use of microorganisms in the synthesis of selected nanoparticles are gaining increased interest as they represent an exciting area of research with considerable development potential. Microorganisms are known to be capable of synthesizing inorganic molecules that are deposited either intra- or extracellularly. This review presents a brief overview of current research on the use of biosurfactants in the biosynthesis of selected metallic nanoparticles and their potential importance. PMID:25110864

  20. Biochar-mediated reductive transformation of nitro herbicides and explosives.

    PubMed

    Oh, Seok-Young; Son, Jong-Gil; Chiu, Pei C

    2013-03-01

    Biochar, a subset of black carbon produced via pyrolysis of biomass, has received much attention in recent years due to its potential to address many important issues, from energy and climate to agriculture and environmental quality. Biochar is known to influence the fate and transport of organic contaminants, although its role has been generally assumed to be as an adsorbent. In this study, the authors investigated the ability of biochar to catalyze the reductive reactions of nitro herbicides and explosives. Two biochars, derived from poultry litter and wastewater biosolids, were found to promote the reductive removal of the dinitro herbicides pendimethalin and trifluralin and the explosives 2,4-dinitrotoluene and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) by dithiothreitol. Parallel experiments using another black carbon material, graphite powder or granular activated carbon, in place of a biochar resulted in comparable rate enhancement to show reduction products, such as 2,4-diaminotoluene and formaldehyde. A cyclization product of trifluralin and reduction products of dinitrotoluene and RDX were detected only when biochar and dithiothreitol were both present, supporting the ability of biochar to promote redox reactions. Three possible catalysts, including graphene moieties, surface functional groups, and redox-active metals, in biochar may be responsible for the biochar-mediated reactions. The environmental significance, implications, and applications of this previously unrecognized role of biochar are discussed. Copyright © 2013 SETAC.

  1. Transformations of metal species in ageing humic hydrocolloids studied by competitive ligand and metal exchange.

    PubMed

    Burba, Peter; Van den Bergh, Johan

    2004-03-01

    Transformations of metal species (particularly Al, Ca, Fe, Mg, Mn, Zn) in ageing humic hydrocolloids were studied, applying a competitive ligand and metal exchange approach. For this purpose, metal-containing hydrocolloids, freshly collected from humic-rich German bog lake waters (Hohlohsee (HO), Black Forest; Venner Moor (VM), Muensterland; Arnsberger Wald (AW), Northrhine-Westfalia) and conventionally pre-filtered through 0.45 microm membranes, were subjected on-site to an exchange with EDTA and Cu(II) ions, respectively, as a function of time. EDTA complexes gradually formed, metal fractions exchanged by Cu(II) (as well as free Cu(II) concentrations) were operationally discriminated by means of a small time-controlled tangential-flow ultrafiltration unit (nominal cutoff: 1 kDa). Metal and DOM (dissolved organic matter) fractions obtained this way were determined off-site using instrumental methods (AAS, ICP-OES, carbon analyzer). After weeks of storage, the collected hydrocolloids were studied again by this approach. The EDTA availability of colloid-bound metals (particularly Al and Fe) exhibited different ageing trends, dependent on the sample (VM: decrease of Fe availability (98-76%), HO: increase of Fe availability (76-82%)). In contrast, the Cu(II) exchange equilibria of colloid-bound metals revealed merely low availability of Al (16-38%) and Fe (5-11%) towards Cu(II) ions, also dependent on ageing effects. In particular, the conditional copper exchange constants Kex obtained from the exchange between Cu(II) ions and available metal species (such as Ca, Mg, Mn, Zn) exhibited a strong decrease (by a factor of 2-100) during sample storage, indicating considerable non-equilibria complexation of these metal ions in the original bogwaters studied on-site.

  2. Metal-Organic Frameworks for CO2 Chemical Transformations.

    PubMed

    He, Hongming; Perman, Jason A; Zhu, Guangshan; Ma, Shengqian

    2016-12-01

    Carbon dioxide (CO2 ), as the primary greenhouse gas in the atmosphere, triggers a series of environmental and energy related problems in the world. Therefore, there is an urgent need to develop multiple methods to capture and convert CO2 into useful chemical products, which can significantly improve the environment and promote sustainable development. Over the past several decades, metal-organic frameworks (MOFs) have shown outstanding heterogeneous catalytic activity due in part to their high internal surface area and chemical functionalities. These properties and the ability to synthesize MOF platforms allow experiments to test structure-function relationships for transforming CO2 into useful chemicals. Herein, recent developments are highlighted for MOFs participating as catalysts for the chemical fixation and photochemical reduction of CO2 . Finally, opportunities and challenges facing MOF catalysts are discussed in this ongoing research area.

  3. Solidification and solid-state transformation sciences in metals additive manufacturing

    DOE PAGES

    Kirka, Michael M.; Nandwana, Peeyush; Lee, Yousub; ...

    2017-02-11

    Additive manufacturing (AM) of metals is rapidly emerging as an established manufacturing process for metal components. Unlike traditional metals fabrication processes, metals fabricated via AM undergo localized thermal cycles during fabrication. As a result, AM presents the opportunity to control the liquid-solid phase transformation, i.e. material texture. But, thermal cycling presents challenges from the standpoint of solid-solid phase transformations. We will discuss the opportunities and challenges in metals AM in the context of texture control and associated solid-solid phase transformations in Ti-6Al-4V and Inconel 718.

  4. Polyethylene glycol (PEG)-mediated transformation in filamentous fungal pathogens

    USDA-ARS?s Scientific Manuscript database

    Genetic transformation is an essential tool in molecular biology for many purposes including the study of gene function and the genetic improvement of an organism. The genetic transformation of many fungal species is a well established process that can be carried out by utilizing different transform...

  5. Polyethylene glycol (PEG)-mediated transformation in filamentous fungal pathogens

    USDA-ARS?s Scientific Manuscript database

    Genetic transformation is an essential tool in molecular biology for many purposes including the study of gene function and the genetic improvement of an organism. The genetic transformation of many fungal species is a well established process that can be carried out by utilizing different transform...

  6. Dislocation mediated alignment during metal nanoparticle coalescence

    DOE PAGES

    Lange, A. P.; Samanta, A.; Majidi, H.; ...

    2016-09-13

    Dislocation mediated alignment processes during gold nanoparticle coalescence were studied at low and high temperatures using molecular dynamics simulations and transmission electron microscopy. Particles underwent rigid body rotations immediately following attachment in both low temperature (500 K) simulated coalescence events and low temperature (~315 K) transmission electron microscopy beam heating experiments. In many low temperature simulations, some degree of misorientation between particles remained after rigid body rotations, which was accommodated by grain boundary dislocation nodes. These dislocations were either sessile and remained at the interface for the duration of the simulation or dissociated and cross-slipped through the adjacent particles, leadingmore » to improved co-alignment. Minimal rigid body rotations were observed during or immediately following attachment in high temperature (1100 K) simulations, which is attributed to enhanced diffusion at the particles' interface. However, rotation was eventually induced by {111} slip on planes parallel to the neck groove. These deformation modes led to the formation of single and multi-fold twins whose structures depended on the initial orientation of the particles. The driving force for {111} slip is attributed to high surface stresses near the intersection of low energy {111} facets in the neck region. The details of this twinning process were examined in detail using simulated trajectories, and the results reveal possible mechanisms for the nucleation and propagation of Shockley partials on consecutive planes. Deformation twinning was also observed in-situ using transmission electron microscopy, which resulted in the co-alignment of a set of the particles' {111} planes across their grain boundary and an increase in their dihedral angle. As a result, this constitutes the first detailed experimental observation of deformation twinning during nanoparticle coalescence, validating simulation results

  7. Dislocation mediated alignment during metal nanoparticle coalescence

    SciTech Connect

    Lange, A. P.; Samanta, A.; Majidi, H.; Mahajan, S.; Ging, J.; Olson, T. Y.; van Benthem, K.; Elhadj, S.

    2016-09-13

    Dislocation mediated alignment processes during gold nanoparticle coalescence were studied at low and high temperatures using molecular dynamics simulations and transmission electron microscopy. Particles underwent rigid body rotations immediately following attachment in both low temperature (500 K) simulated coalescence events and low temperature (~315 K) transmission electron microscopy beam heating experiments. In many low temperature simulations, some degree of misorientation between particles remained after rigid body rotations, which was accommodated by grain boundary dislocation nodes. These dislocations were either sessile and remained at the interface for the duration of the simulation or dissociated and cross-slipped through the adjacent particles, leading to improved co-alignment. Minimal rigid body rotations were observed during or immediately following attachment in high temperature (1100 K) simulations, which is attributed to enhanced diffusion at the particles' interface. However, rotation was eventually induced by {111} slip on planes parallel to the neck groove. These deformation modes led to the formation of single and multi-fold twins whose structures depended on the initial orientation of the particles. The driving force for {111} slip is attributed to high surface stresses near the intersection of low energy {111} facets in the neck region. The details of this twinning process were examined in detail using simulated trajectories, and the results reveal possible mechanisms for the nucleation and propagation of Shockley partials on consecutive planes. Deformation twinning was also observed in-situ using transmission electron microscopy, which resulted in the co-alignment of a set of the particles' {111} planes across their grain boundary and an increase in their dihedral angle. As a result, this constitutes the first detailed experimental observation of deformation twinning during nanoparticle coalescence, validating simulation results

  8. Dislocation mediated alignment during metal nanoparticle coalescence

    SciTech Connect

    Lange, A. P.; Samanta, A.; Majidi, H.; Mahajan, S.; Ging, J.; Olson, T. Y.; van Benthem, K.; Elhadj, S.

    2016-09-13

    Dislocation mediated alignment processes during gold nanoparticle coalescence were studied at low and high temperatures using molecular dynamics simulations and transmission electron microscopy. Particles underwent rigid body rotations immediately following attachment in both low temperature (500 K) simulated coalescence events and low temperature (~315 K) transmission electron microscopy beam heating experiments. In many low temperature simulations, some degree of misorientation between particles remained after rigid body rotations, which was accommodated by grain boundary dislocation nodes. These dislocations were either sessile and remained at the interface for the duration of the simulation or dissociated and cross-slipped through the adjacent particles, leading to improved co-alignment. Minimal rigid body rotations were observed during or immediately following attachment in high temperature (1100 K) simulations, which is attributed to enhanced diffusion at the particles' interface. However, rotation was eventually induced by {111} slip on planes parallel to the neck groove. These deformation modes led to the formation of single and multi-fold twins whose structures depended on the initial orientation of the particles. The driving force for {111} slip is attributed to high surface stresses near the intersection of low energy {111} facets in the neck region. The details of this twinning process were examined in detail using simulated trajectories, and the results reveal possible mechanisms for the nucleation and propagation of Shockley partials on consecutive planes. Deformation twinning was also observed in-situ using transmission electron microscopy, which resulted in the co-alignment of a set of the particles' {111} planes across their grain boundary and an increase in their dihedral angle. As a result, this constitutes the first detailed experimental observation of deformation twinning during nanoparticle coalescence, validating simulation results

  9. Redox-mediated activation of latent transforming growth factor-beta 1

    NASA Technical Reports Server (NTRS)

    Barcellos-Hoff, M. H.; Dix, T. A.; Chatterjee, A. (Principal Investigator)

    1996-01-01

    Transforming growth factor beta 1 (TGF beta) is a multifunctional cytokine that orchestrates response to injury via ubiquitous cell surface receptors. The biological activity of TGF beta is restrained by its secretion as a latent complex (LTGF beta) such that activation determines the extent of TGF beta activity during physiological and pathological events. TGF beta action has been implicated in a variety of reactive oxygen-mediated tissue processes, particularly inflammation, and in pathologies such as reperfusion injury, rheumatoid arthritis, and atherosclerosis. It was recently shown to be rapidly activated after in vivo radiation exposure, which also generates reactive oxygen species (ROS). In the present studies, the potential for redox-mediated LTGF beta activation was investigated using a cell-free system in which ROS were generated in solution by ionizing radiation or metal ion-catalyzed ascorbate reaction. Irradiation (100 Gray) of recombinant human LTGF beta in solution induced 26% activation compared with that elicited by standard thermal activation. Metal-catalyzed ascorbate oxidation elicited extremely efficient recombinant LTGF beta activation that matched or exceeded thermal activation. The efficiency of ascorbate activation depended on ascorbate concentrations and the presence of transition metal ions. We postulate that oxidation of specific amino acids in the latency-conferring peptide leads to a conformation change in the latent complex that allows release of TGF beta. Oxidative activation offers a novel route for the involvement of TGF beta in tissue processes in which ROS are implicated and endows LTGF beta with the ability to act as a sensor of oxidative stress and, by releasing TGF beta, to function as a signal for orchestrating the response of multiple cell types. LTGF beta redox sensitivity is presumably directed toward recovery of homeostasis; however, oxidation may also be a mechanism of LTGF beta activation that can be deleterious during

  10. Redox-mediated activation of latent transforming growth factor-beta 1

    NASA Technical Reports Server (NTRS)

    Barcellos-Hoff, M. H.; Dix, T. A.; Chatterjee, A. (Principal Investigator)

    1996-01-01

    Transforming growth factor beta 1 (TGF beta) is a multifunctional cytokine that orchestrates response to injury via ubiquitous cell surface receptors. The biological activity of TGF beta is restrained by its secretion as a latent complex (LTGF beta) such that activation determines the extent of TGF beta activity during physiological and pathological events. TGF beta action has been implicated in a variety of reactive oxygen-mediated tissue processes, particularly inflammation, and in pathologies such as reperfusion injury, rheumatoid arthritis, and atherosclerosis. It was recently shown to be rapidly activated after in vivo radiation exposure, which also generates reactive oxygen species (ROS). In the present studies, the potential for redox-mediated LTGF beta activation was investigated using a cell-free system in which ROS were generated in solution by ionizing radiation or metal ion-catalyzed ascorbate reaction. Irradiation (100 Gray) of recombinant human LTGF beta in solution induced 26% activation compared with that elicited by standard thermal activation. Metal-catalyzed ascorbate oxidation elicited extremely efficient recombinant LTGF beta activation that matched or exceeded thermal activation. The efficiency of ascorbate activation depended on ascorbate concentrations and the presence of transition metal ions. We postulate that oxidation of specific amino acids in the latency-conferring peptide leads to a conformation change in the latent complex that allows release of TGF beta. Oxidative activation offers a novel route for the involvement of TGF beta in tissue processes in which ROS are implicated and endows LTGF beta with the ability to act as a sensor of oxidative stress and, by releasing TGF beta, to function as a signal for orchestrating the response of multiple cell types. LTGF beta redox sensitivity is presumably directed toward recovery of homeostasis; however, oxidation may also be a mechanism of LTGF beta activation that can be deleterious during

  11. Ecotoxic heavy metals transformation by bacteria and fungi in aquatic ecosystem.

    PubMed

    Chaturvedi, Amiy Dutt; Pal, Dharm; Penta, Santhosh; Kumar, Awanish

    2015-10-01

    Water is the most important and vital molecule of our planet and covers 75% of earth surface. But it is getting polluted due to high industrial growth. The heavy metals produced by industrial activities are recurrently added to it and considered as dangerous pollutants. Increasing concentration of toxic heavy metals (Pb(2+), Cd(2+), Hg(2+), Ni(2+)) in water is a severe threat for human. Heavy metal contaminated water is highly carcinogenic and poisonous at even relatively low concentrations. When they discharged in water bodies, they dissolve in the water and are distributed in the food chain. Bacteria and fungi are efficient microbes that frequently transform heavy metals and remove toxicity. The application of bacteria and fungi may offer cost benefit in water treatment plants for heavy metal transformation and directly related to public health and environmental safety issues. The heavy metals transformation rate in water is also dependent on the enzymatic capability of microorganisms. By transforming toxic heavy metals microbes sustain aquatic and terrestrial life. Therefore the application of microbiological biomass for heavy metal transformation and removal from aquatic ecosystem is highly significant and striking. This paper reviews the microbial transformation of heavy metal, microbe metal interaction and different approaches for microbial heavy metal remediation from water bodies.

  12. Phytochemicals Mediated Remediation of Neurotoxicity Induced by Heavy Metals.

    PubMed

    Gupta, Vivek Kumar; Singh, Shweta; Agrawal, Anju; Siddiqi, Nikhat Jamal; Sharma, Bechan

    2015-01-01

    Almost all the environmental components including both the abiotic and biotic factors have been consistently threatened by excessive contamination of heavy metals continuously released from various sources. Different heavy metals have been reported to generate adverse effects in many ways. Heavy metals induced neurotoxicity and impairment in signalling cascade leading to cell death (apoptosis) has been indicated by several workers. On one hand, these metals are required by the cellular systems to regulate various biological functions of normal cells, while on the other their biomagnification in the cellular systems produces adverse effects. The mechanism by which the heavy metals induce neurotoxicity follows free radicals production pathway(s) specially the generation of reactive oxygen species and reactive nitrogen species. These free radicals produced in excess have been shown to create an imbalance between the oxidative and antioxidative systems leading to emergence of oxidative stress, which may cause necrosis, DNA damage, and many neurodegenerative disorders. This mini review summarizes the current knowledge available on the protective role of varied natural products isolated from different herbs/plants in imparting protection against heavy metals (cadmium, lead, arsenic, and mercury) mediated neurotoxicity.

  13. Phytochemicals Mediated Remediation of Neurotoxicity Induced by Heavy Metals

    PubMed Central

    Gupta, Vivek Kumar; Singh, Shweta; Agrawal, Anju; Siddiqi, Nikhat Jamal; Sharma, Bechan

    2015-01-01

    Almost all the environmental components including both the abiotic and biotic factors have been consistently threatened by excessive contamination of heavy metals continuously released from various sources. Different heavy metals have been reported to generate adverse effects in many ways. Heavy metals induced neurotoxicity and impairment in signalling cascade leading to cell death (apoptosis) has been indicated by several workers. On one hand, these metals are required by the cellular systems to regulate various biological functions of normal cells, while on the other their biomagnification in the cellular systems produces adverse effects. The mechanism by which the heavy metals induce neurotoxicity follows free radicals production pathway(s) specially the generation of reactive oxygen species and reactive nitrogen species. These free radicals produced in excess have been shown to create an imbalance between the oxidative and antioxidative systems leading to emergence of oxidative stress, which may cause necrosis, DNA damage, and many neurodegenerative disorders. This mini review summarizes the current knowledge available on the protective role of varied natural products isolated from different herbs/plants in imparting protection against heavy metals (cadmium, lead, arsenic, and mercury) mediated neurotoxicity. PMID:26618004

  14. Phosphate-Mediated Remediation of Metals and Radionuclides

    DOE PAGES

    Martinez, Robert J.; Beazley, Melanie J.; Sobecky, Patricia A.

    2014-01-01

    Worldwide industrialization activities create vast amounts of organic and inorganic waste streams that frequently result in significant soil and groundwater contamination. Metals and radionuclides are of particular concern due to their mobility and long-term persistence in aquatic and terrestrial environments. As the global population increases, the demand for safe, contaminant-free soil and groundwater will increase as will the need for effective and inexpensive remediation strategies. Remediation strategies that include physical and chemical methods (i.e., abiotic) or biological activities have been shown to impede the migration of radionuclide and metal contaminants within soil and groundwater. However, abiotic remediation methods aremore » often too costly owing to the quantities and volumes of soils and/or groundwater requiring treatment. The in situ sequestration of metals and radionuclides mediated by biological activities associated with microbial phosphorus metabolism is a promising and less costly addition to our existing remediation methods. This review highlights the current strategies for abiotic and microbial phosphate-mediated techniques for uranium and metal remediation.« less

  15. Chemically and temperature-induced phase transformations of metal vanadates

    NASA Astrophysics Data System (ADS)

    Patridge, Christopher James

    different individual beta'-Cu xV2O5 nanowires vary widely. Using scanning transmission X-ray microspectroscopy of individual beta'-CuxV2O 5 nanowires, correlations appear to exist between MIT characteristics and the markedly different orbital hybridization of vanadium and oxygen at the O K and V L absorption edges. These comprehensive nanostructure studies hint at the possibility of approaching the incredibly important realm of single-domain measurements which are needed to understand and exploit the intrinsic physical properties of materials. In addition to the bronze MIT studies, the classical MIT material vanadium dioxide, VO2, also shows new properties when scaling down to nanoscale dimensions as well as incorporation of substitutional dopants such as tungsten. X-ray absorption spectroscopy of the dopant local structure suggests an increased symmetry and depairing of V4+-V 4+, which is critical for transition to the lower temperature insulating phase thereby super-cooling the metallic phase to temperatures as low as 254 K. Mechanistic insight and structural changes associated with the intercalation of Li+ are key aspects in understanding and designing useful secondary Li ion batteries. In similarity to the MxV2O 5 studies, another metal vanadate, Ag2VO2PO 4, undergoes phase transformations due to introduction of Li and the vacancy of Ag ions. Employing a comprehensive study on Ag2VO 2PO4 using X-ray absorption spectroscopy, information about chemical state changes and rehybridization of frontier orbitals allows for a more precise understanding of how the material discharges, what, if any, intermediate phases exist during the process, and provides evidence for the posited structural stability at high depths of discharge.

  16. Actinomycetes mediated biogenic synthesis of metal and metal oxide nanoparticles: Progress and challenges.

    PubMed

    Manimaran, Manickavelu; Kannabiran, Krishnan

    2017-03-07

    Actinomycetes mediated biogenic synthesis of metal nanoparticles and their antimicrobial activities are well documented. Actinomycetes facilitate both intracellular and extracellular metal nanoparticles synthesis and are efficient candidates for the production of polydispersed, stable and ultra-small size metal nanoparticles. Secondary metabolites and new chemical entities derived from actinomycetes have not been extensively studied for the synthesis of metal/ metal oxide nanoparticles. The present review focuses on biogenic synthesis of metal nanoparticles from actinomycetes and the scope for exploring actinomycetes derived compounds (enzymes, organics acids and bioactive compounds) as metal and metal oxide reducing agents for the synthesis of desired nanoparticles. This review also focuses on challenges faced in the applications nanoparticles and the methods to synthesise biogenic metal nanoparticles with desired physiochemical properties such as ultra-small size, large surface to mass ratio, high reactivity etc. Methods to evade their toxicity and unique interactions with biological systems to improve their chance as an alternative therapeutic agent in medical and pharmaceutical industry are also discussed. This article is protected by copyright. All rights reserved.

  17. Transformation of tetracyclines mediated by Mn(II) and Cu(II) ions in the presence of oxygen.

    PubMed

    Chen, Wan-Ru; Huang, Ching-Hua

    2009-01-15

    Complexation of tetracyclines (TCs) with dissolved Mn(II) and Cu(II) ions were found to significantly enhance the transformation of these antibiotics in the presence of oxygen at pH 8-9.5 and pH 4-6, respectively. In the TC-Mn(II)-O2 system, oxidation of the TC-complexed Mn(II) to Mn(III) by oxygen occurs, followed by oxidation of TC by Mn(III) to regenerate Mn(II). In the TC-Cu(II)-O2 system, Cu(II) oxidizes TC within the complex and the yielded Cu(I) is reoxidized by the present oxygen. Opposite reactivity trends were observed with the two metals: OTC (oxytetracycline) > TTC (tetracycline) > iso-CTC (iso-chlorotetracycline) for the Mn(II)-mediated reaction, whereas CTC > TTC > OTC > epimers for the Cu(II)-mediated reaction. The reactivity results and examination of TC-metal ion complexation and transformation products suggest that the BCD-ring and A-ring of TC are crucial to interact with Mn(II) and Cu(II), respectively. This study highlights that the fate of TCs in aquatic environments may differ significantly by their strong interactions with different metal species present in the systems.

  18. Phytoremediation using microbially mediated metal accumulation in Sorghum bicolor.

    PubMed

    Phieler, René; Merten, Dirk; Roth, Martin; Büchel, Georg; Kothe, Erika

    2015-12-01

    Reclaiming land that has been anthropogenically contaminated with multiple heavy metal elements, e.g., during mining operations, is a growing challenge worldwide. The use of phytoremediation has been discussed with varying success. Here, we show that a careful examination of options of microbial determination of plant performance is a key element in providing a multielement remediation option for such landscapes. We used both (a) mycorrhiza with Rhizophagus irregularis and (b) bacterial amendments with Streptomyces acidiscabies E13 and Streptomyces tendae F4 to mediate plant-promoting and metal-accumulating properties to Sorghum bicolor. In pot experiments, the effects on plant growth and metal uptake were scored, and in a field trial at a former uranium leaching heap site near Ronneburg, Germany, we could show the efficacy under field conditions. Different metals could be extracted at the same time, with varying microbial inoculation and soil amendment scenarios possible when a certain metal is the focus of interest. Especially, manganese was extracted at very high levels which might be useful even for phytomining approaches.

  19. Agrobacterium tumefaciens-mediated transformation of Narcissus tazzeta var. chinensis.

    PubMed

    Lu, Gang; Zou, Qingcheng; Guo, Deping; Zhuang, Xiaoying; Yu, Xiaolin; Xiang, Xun; Cao, Jiashu

    2007-09-01

    Phytoene synthase (PSY), as a key regulatory enzyme for carotene biosynthesis, plays an important role in regulating color formation in many species. In the present study, a protocol was developed for the transformation of Narcissus tazzeta var chinensis using Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pCAMBIA1301 plasmid which contained an antisense phytoene synthase gene, a reporter beta-glucuronidase gene and a selectable marker hygromycin phosphotransferase gene. Effects of some factors on efficiency of transformation and regeneration were examined. Preculture of the explants for 6 days before inoculation enhanced the transient GUS expression. The addition of acetosyringone (AS) at 100 micromol l(-1) for inoculation and a period of 3 days co-cultivation yielded efficient transient GUS expression. Transformants were obtained through selection on MS medium containing 5 mg l(-1) 6-benzylaminopurine (BA), 0.1 mg l(-1)alpha-naphthalene acetic acid (NAA) and 40 mg l(-1) hygromycin. The transformation frequency was 1.24% based on PCR analysis of gus gene. One or two copies of transgene were demonstrated in different transformations by Southern blotting analyses. Northern blotting results confirmed that the transcription of the endogenous psy gene in transgenic plants was inhibited or silenced. The method reported here provides new opportunities for improvement of quality traits of Narcissus tazzeta via genetic transformation.

  20. Agrobacterium-mediated genetic transformation and plant regeneration of the hardwood tree species Fraxinus profunda

    Treesearch

    Micah E. Stevens; Paula M. Pijut

    2014-01-01

    Using mature hypocotyls as the initial explants, an Agrobacterium tumefaciens-mediated genetic transformation system was successfully developed for pumpkin ash (Fraxinus profunda). This transformation protocol is an invaluable tool to combat the highly aggressive, non-native emerald ash borer (EAB), which has the potential to...

  1. Improvement of Agrobacterium-mediated transformation and rooting of black cherry

    Treesearch

    Ying Wang; Paula M. Pijut

    2014-01-01

    An improved protocol for Agrobacterium-mediated transformation of an elite, mature black cherry genotype was developed. To increase transformation efficiency, vacuum infiltration, sonication, and a combination of the two treatments were applied during the cocultivation of leaf explants with Agrobacterium tumefaciens strain EHA105...

  2. Phase transformations and thermodynamics of aluminum-based metallic glasses

    NASA Astrophysics Data System (ADS)

    Gao, Changhua (Michael)

    This thesis examines the thermodynamics and associated kinetics and phase transformations of the glass forming Al-Ni-Gd and Al-Fe-Gd systems. In order to fully understand the unique glass forming ability (GFA) of Al-based metallic glasses, the ternary Al-Fe-Gd and Al-Ni-Gd systems in their Al-rich corners were examined experimentally to assist in a thermodynamic assessment. The solid-state phase equilibria are determined using XRD and TEM-EDS techniques. While this work basically confirms the solid-state equilibria in Al-Fe-Gd reported previously, the ternary phase in Al-Ni-Gd system has been identified to be Al15Ni3Gd2 rather than Al16Ni 3Gd reported in the literature. DTA analysis of 24 alloys in the Al-Fe-Gd system and 42 alloys in the Al-Ni-Gd system have yielded critical temperatures pertaining to the solid-liquid transition. Based on these data and information from the literature, a self-consistent thermodynamic database for these systems has been developed using the CALPHAD technique. Parameters describing the Gibbs free energy for various phases of the Al-Gd, Al-Fe-Gd and Al-Ni-Gd systems are manually optimized in this study. Once constructed, the database is used to calculate driving forces for nucleation of crystalline phases which can qualitatively explain the phase formation sequence during crystallization at low temperatures. It was also confirmed that alloy compositions with the lowest Gibbs free energy difference between the equilibrium state and undercooled liquid state exhibit better GFA than other chemistries. Based on 250°C isothermal devitrification phase transformations of 17 Al-Ni-Gd alloys, a phase formation sequence map is constructed. Fcc-Al nanocrystals are formed first in most of the alloys studied, but eutectic crystallization of a metastable phase and fcc-Al is also observed. Addition of Al or Ni promotes fcc-Al phase formation, while increasing Gd suppresses it. The continuous heating DSC scans revealed that crystallization in Al

  3. Transformational Leadership and Teacher Commitment to Organizational Values: The Mediating Effects of Collective Teacher Efficacy

    ERIC Educational Resources Information Center

    Ross, John A.; Gray, Peter

    2006-01-01

    Transformational leadership researchers have given little attention to teacher expectations that mediate between goals and actions. The most important of these expectations, teacher efficacy, refers to teacher beliefs that they will be able to bring about student learning. This study examined the mediating effects of teacher efficacy by comparing…

  4. Distinct tyrosine autophosphorylation sites negatively and positively modulate neu-mediated transformation.

    PubMed Central

    Dankort, D L; Wang, Z; Blackmore, V; Moran, M F; Muller, W J

    1997-01-01

    A number of cytoplasmic signaling molecules are thought to mediate mitogenic signaling from the activated Neu receptor tyrosine kinase through binding specific phosphotyrosine residues located within the intracellular portion of Neu/c-ErbB-2. An activated neu oncogene containing tyrosine-to-phenylalanine substitutions at each of the known autophosphorylation sites was generated and assessed for its specific transforming potential in Rat1 and NIH 3T3 fibroblasts. Mutation of these sites resulted in a dramatic impairment of the transforming potential of neu. To assess the role of these tyrosine phosphorylation sites in cellular transformation, the transforming potential of a series of mutants in which individual tyrosine residues were restored to this transformation-debilitated neu mutant was evaluated. Reversion of any one of four mutated sites to tyrosine residues restored wild-type transforming activity. While each of these transforming mutants displayed Ras-dependent signaling, the transforming activity of two of these mutants was correlated with their ability to bind either the GRB2 or SHC adapter molecules that couple receptor tyrosine kinases to the Ras signaling pathway. By contrast, restoration of a tyrosine residue located at position 1028 completely suppressed the basal transforming activity of this mutated neu molecule or other transforming neu molecules which possessed single tyrosine residues. These data argue that the transforming potential of activated neu is mediated both by positive and negative regulatory tyrosine phosphorylation sites. PMID:9271418

  5. ipt Gene transformation in petunia by an Agrobacterium mediated method.

    PubMed

    Bai, L J; Ye, C J; Lu, J Y; Yang, D E; Xue, H; Pan, Y; Cao, P X; Wang, B; Liu, M

    2009-01-01

    To prevent leaf senescence of petunia, the cytokinin biosynthetic gene isopentenyl transferase (ipt) was placed under the control of 35S promoter and introduced into petunia. PCR analysis showed an expected 0.5 Kb fragment of ipt gene in transgenic petunia. RT-PCR analysis indicated the expression of ipt gene in the transgenic lines. Leaves from transgenic plants remained green and healthy in normal culture condition, while the non-transformed plants turned to yellow. Transgenic plants showed a reduction in height and smaller leaf sizes. In transgenic lines, the internodes were shorter, and the roots grew slower than the non-transformed plants.

  6. Enhanced transformation of triclosan by laccase in the presence of redox mediators.

    PubMed

    Murugesan, Kumarasamy; Chang, Yoon-Young; Kim, Young-Mo; Jeon, Jong-Rok; Kim, Eun-Ju; Chang, Yoon-Seok

    2010-01-01

    Triclosan (TCS), an antimicrobial agent, is an emerging and persistent environmental pollutant that is often found as a contaminant in surface waters and sediments; hence, knowledge of its degradability is important. In this study we investigated laccase-mediated TCS transformation and detoxification, using laccase (from the fungus Ganoderma lucidum) in the presence and absence of redox mediators. Transformation products were identified using HPLC, ESI-MS and GC-MS, and transformation mechanisms were proposed. In the absence of redox mediator, 56.5% TCS removal was observed within 24h, concomitant with formation of new products with molecular weights greater than that of TCS. These products were dimers and trimers of TCS, as confirmed by ESI-MS analysis. Among the various mediators tested, 1-hydroxybenzotriazole (HBT) and syringaldehyde (SYD) significantly enhanced TCS transformation ( approximately 90%). The presence of these mediators resulted in products with lower molecular weights than TCS, including 2,4-dichlorophenol (2,4-DCP; confirmed by GC-MS) and dechlorinated forms of 2,4-DCP. When SYD was used as the mediator, dechlorination resulted in 2-chlorohydroquinone (2-CHQ). Bacterial growth inhibition studies revealed that laccase-mediated transformation of TCS effectively decreased its toxicity, with ultimate conversion to less toxic or nontoxic products. Our results confirmed the involvement of two mechanisms of laccase-catalyzed TCS removal: (i) oligomerization in the absence of redox mediators, and (ii) ether bond cleavage followed by dechlorination in the presence of redox mediators. These results suggest that laccase in combination with natural redox mediator systems may be a useful strategy for the detoxification and elimination of TCS from aqueous systems.

  7. Solution-mediated phase transformation: significance during dissolution and implications for bioavailability.

    PubMed

    Greco, Kristyn; Bogner, Robin

    2012-09-01

    Solubility improvement of poorly soluble drug compounds is a key approach to ensuring the successful development of many new drugs. Methods used to improve the solubility of drug compounds include forming a salt, cocrystal, or amorphous solid. These methods of improving solubility can often lead to a phenomenon called solution-mediated phase transformation, a phase change that is facilitated through exposure to solution. Solution-mediated phase transformation occurs in three steps: dissolution to create a supersaturated solution followed by nucleation of less soluble phase and the growth of that phase. When the growth of the less soluble phase occurs on the surface of the metastable solid, this phenomenon can cause a marked decrease in dissolution rate during in vitro dissolution evaluation, and ultimately in vivo. Therefore, transformation to a less soluble solid during dissolution is an important aspect to consider when evaluating approaches to increase the solubility of a poorly soluble drug. Identification of solution-mediated phase transformation during dissolution is reviewed for powder dissolution, rotating disk method, and channel flow-through apparatus. Types of solution-mediated phase transformation are described in this report, including those involving salts, polymorphs, amorphous solids, and cocrystals. Many experimental examples are provided. Evidence of potential solution-mediated phase transformation in vivo is discussed to better understand the relationship between in vitro dissolution evaluation and in vivo performance.

  8. Transformational leadership and task cohesion in sport: the mediating role of inside sacrifice.

    PubMed

    Cronin, Lorcan Donal; Arthur, Calum Alexander; Hardy, James; Callow, Nichola

    2015-02-01

    In this cross-sectional study, we examined a mediational model whereby transformational leadership is related to task cohesion via sacrifice. Participants were 381 American (Mage = 19.87 years, SD = 1.41) Division I university athletes (188 males, 193 females) who competed in a variety of sports. Participants completed measures of coach transformational leadership, personal and teammate inside sacrifice, and task cohesion. After conducting multilevel mediation analysis, we found that both personal and teammate inside sacrifice significantly mediated the relationships between transformational leadership behaviors and task cohesion. However, there were differential patterns of these relationships for male and female athletes. Interpretation of the results highlights that coaches should endeavor to display transformational leadership behaviors as they are related to personal and teammate inside sacrifices and task cohesion.

  9. Steric Switching from Photochemical to Thermal Reaction Pathways for Enhanced Efficiency in Metal-Mediated Nitrogen Fixation.

    PubMed

    Duman, Leila M; Farrell, Wesley S; Zavalij, Peter Y; Sita, Lawrence R

    2016-11-16

    Programmed manipulation of the subtle interplay of nonbonded steric interactions within a supporting ligand environment has been used for the conversion of a photochemically driven chemical cycle for group 6 metal-mediated nitrogen fixation into a thermally promoted process with increased energy efficiency and atom economy for key transformations involving N≡N bond cleavage and N-atom functionalization of coordinated N2.

  10. Interrupting Base-Mediated Benzofuran Ring Transformation with Michael Acceptors.

    PubMed

    Srinivas, Kolluru; Sharma, Rashmi; Ramana, Chepuri V

    2017-08-29

    A simple two-stage approach for the synthesis of 3-(2-arylbenzofuran-3-yl)propanoates and propanamides has been developed employing simple acrylates and acrylamides and readily available 3-aroylbenzofurans. The key step of this process involves a base-mediated ring opening of the 3-aroylbenzofurans and subsequent Michael addition of the resulting 1,3-dicarbonyl intermediate with acrylate/acrylamide, followed by the deformylation in one-pot. The resulting products undergo an acid-mediated dehydrative cyclization to arrive at these targets.

  11. UV stimulation of DNA-mediated transformation of human cells

    SciTech Connect

    van Duin, M.; Westerveld, A.; Hoeijmakers, J.H.

    1985-04-01

    Irradiation of dominant marker DNA with UV light (150 to 1,000 J/m2) was found to stimulate the transformation of human cells by this marker from two- to more than fourfold. This phenomenon is also displayed by xeroderma pigmentosum cells, which are deficient in the excision repair of UV-induced pyrimidine dimers in the DNA. Also, exposure to UV of the transfected (xeroderma pigmentosum) cells enhanced the transfection efficiency. Removal of the pyrimidine dimers from the DNA by photoreactivating enzyme before transfection completely abolished the stimulatory effect, indicating that dimer lesions are mainly responsible for the observed enhancement. A similar stimulation of the transformation efficiency is exerted by 2-acetoxy-2-acetylaminofluorene modification of the DNA. These findings suggest that lesions which are targets for the excision repair pathway induce the increase in transformation frequency. The stimulation was found to be independent of sequence homology between the irradiated DNA and the host chromosomal DNA. Therefore, the increase of the transformation frequency is not caused by a mechanism inducing homologous recombination between these two DNAs. UV treatment of DNA before transfection did not have a significant effect on the amount of DNA integrated into the xeroderma pigmentosum genome.

  12. Mediating the Conflict between Transformative Pedagogy and Bureaucratic Practice

    ERIC Educational Resources Information Center

    Inderbitzin, Michelle; Storrs, Debbie A.

    2008-01-01

    This article reflects on the authors' experiences during a pilot year of an innovative core curriculum at a state research university and their attempts to create a "collaborative community" characterized by transformative pedagogy. It discusses their students' and colleagues' resistance to their inventive, albeit time-consuming and sometimes…

  13. Transforming Ourselves through the Power of Mediated Instruction.

    ERIC Educational Resources Information Center

    Guadarrama, Irma N., Ed.; Kirksey, Lockie, Ed.

    1996-01-01

    A collection of essays on English-as-a-Second-Language (ESL) and bilingual education focuses on issues in making curricula meaningful for teachers and students. Articles include: "Critical Mediation: When Teachers and Students Connect in the 'Ecliptic Zone'" (Irma N. Guadarrama); "Reflecting on Ideological Baggage: Latino…

  14. Transforming Ourselves through the Power of Mediated Instruction.

    ERIC Educational Resources Information Center

    Guadarrama, Irma N., Ed.; Kirksey, Lockie, Ed.

    1996-01-01

    A collection of essays on English-as-a-Second-Language (ESL) and bilingual education focuses on issues in making curricula meaningful for teachers and students. Articles include: "Critical Mediation: When Teachers and Students Connect in the 'Ecliptic Zone'" (Irma N. Guadarrama); "Reflecting on Ideological Baggage: Latino…

  15. Role of bacterial virulence proteins in Agrobacterium-mediated transformation of Aspergillus awamori.

    PubMed

    Michielse, C B; Ram, A F J; Hooykaas, P J J; Hondel, C A M J J van den

    2004-05-01

    The Agrobacterium-mediated transformation of Aspergillus awamori was optimized using defined co-cultivation conditions, which resulted in a reproducible and efficient transformation system. Optimal co-cultivation conditions were used to study the role of Agrobacterium tumefaciens virulence proteins in T-DNA transfer. This study revealed that inactivation of either of the regulatory proteins (VirA, VirG), any of the transport pore proteins (VirB), proteins involved in generation of the T-strand (VirD, VirC) or T-strand protection and targeting (VirE2) abolishes or severely reduces the formation of transformants. The results indicate that the Agrobacterium-mediated transformation of A. awamori requires an intact T-DNA machinery for efficient transformation; however, the plant host range factors, like VirE3, VirH, and VirF, are not important.

  16. PHASE TRANSFORMATIONS IN METALS AND ALLOYS (SELECTED ARTICLES),

    DTIC Science & Technology

    Contents: Stabilization of reverse martensite transformation under the influence of intraphase work hardening; and Structural changes during decomposition of supersaturated solid solution of tungsten in cobalt.

  17. Agrobacterium-mediated transformation of Ruta graveolens L.

    PubMed

    Lièvre, Karine; Tran, Thi Lê Minh; Doerper, Sébastien; Hehn, Alain; Lacoste, Paul; Thomasset, Brigitte; Bourgaud, Frédéric; Gontier, Eric

    2009-01-01

    Agrobacterium tumefaciens is used to develop a genetic transformation method for a medicinal plant Ruta graveolens. The direct plant regeneration strategy is preferred to callus line establishment. In vitro seedlings, 2- -to 3-wk-old, are used to excise hypocotyls and co-cultivated for 3 d with A. tumefaciens strain C58C1Rif containing plasmid pTDE4 harbouring neomycin phosphotransferase (npt II, kanamycin resistance) and beta-glucuronidase encoding genes. The Southern blot analysis has shown that 78% kanamycin resistant plants contain gene encoding beta-glucuronidase. The GUS histochemical assay shows that 67% transgenic plants exhibit the corresponding enzymatic activity. Routine transformation efficiency of R. graveolens L. is 11% and could reach up to 22%. Transgenic plants are grown in the greenhouse within 4 months after the initial seedlings.

  18. Simulating the transformation of heavy metals during coal or sewage sludge combustion

    SciTech Connect

    Han, J.; Xu, M.; Yao, H.; Furuuchi, M.; Sakano, T.; Kim, H.J.

    2007-01-15

    A mathematical model (FPM) is presented to predict the transformation of heavy metals in the downstream of combustor or incinerator. The model accounts for the transformation of heavy metals through the combined effect of condensation, nucleation, coagulation, external force and thermophoresis force. The calculation of heavy metals is embodied in the post-processor appended to Fluent software. Before the simulation, velocity, temperature, PbCl{sub 2} concentration and other initial parameters are obtained by experiment. In addition, the transformation of PbCl{sub 2} is also experimentally studied. The comparison of experimental and predicted results indicate that the fine particle model (FPM) is valid for predicting the transformation of heavy metals in the downstream of incinerator or combustor.

  19. Migration and transformation rule of heavy metals in sludge during hydrolysis for protein extraction.

    PubMed

    Li, Yulong; Xue, Fei; Li, Jiebing; Xu, Shi Hong; Li, Dengxin

    2016-03-01

    The content and speciation of heavy metals can fundamentally affect the hydrolysis of sludge. This research study investigates the migration and transformation rule of heavy metals during the hydrolysis process by measuring the content of exchangeables (F1), bound to carbonates (F2), bound to Fe-Mn oxides (F3), bound to organic matter (F4), and residuals (F5) under different periods of time undergoing hydrolysis. The results show that the hydrolysis process generally stabilized Cu, Zn, Mn, Ni, Pb, Cr, and As by transforming the unstable states into structurally stable states. Such transformations and stabilization were primarily caused by the changes in local metal ion environment and bonding structure, oxidation of sulfides, pyrolyzation of organic matter, and evaporation of resulting volatile materials. An X-ray diffractometry (XRD) of the residuals conducted after hydrolysis indicated that hydrolysis did have a significant influence on the transportation and transformation of heavy metals.

  20. Recent Developments in Coinage Metal Catalyzed Transformations of Stabilized Vinyldiazo Compounds: Beyond Carbenic Pathways.

    PubMed

    López, Enol; González-Pelayo, Silvia; López, Luis A

    2017-03-01

    Transition metal-catalyzed transformations of vinyldiazo compounds have become a versatile tool in organic synthesis. Although several transition metals have been investigated for this purpose, this field has been mainly dominated by dirhodium catalysts. Remarkable levels of chemo-, regio-, diastereo- and enantioselectivity have been reached in some of these rhodium-catalyzed transformations. In the last few years coinage metals have also emerged as useful catalysts in transformations involving vinyldiazo compounds. In some cases, highly efficient catalyst-dependent protocols arising from divergent mechanistic pathways have been reported. In this Personal Account, we aim to showcase recent advances in metal coinage catalyzed transformations of vinyldiazoacetates, an exciting field of research to which our group has actively contributed in the last few years.

  1. Development of Transgenic Papaya through Agrobacterium-Mediated Transformation

    PubMed Central

    Azad, Md. Abul Kalam; Rabbani, Md. Golam; Amin, Latifah; Sidik, Nik Marzuki

    2013-01-01

    Transgenic papaya plants were regenerated from hypocotyls and immature zygotic embryo after cocultivation with Agrobacterium tumefaciens LBA-4404 carrying a binary plasmid vector system containing neomycin phosphotransferase (nptII) gene as the selectable marker and β-glucuronidase (GUS) as the reporter gene. The explants were co-cultivated with Agrobacterium tumefaciens on regeneration medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime for one week. The cocultivated explants were transferred into the final selection medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime + 50 mg/L kanamycin for callus induction as well as plant regeneration. The callus derived from the hypocotyls of Carica papaya cv. Shahi showed the highest positive GUS activities compared to Carica papaya cv. Ranchi. The transformed callus grew vigorously and formed embryos followed by transgenic plantlets successfully. The result of this study showed that the hypocotyls of C. papaya cv. Shahi and C. papaya cv. Ranchi are better explants for genetic transformation compared to immature embryos. The transformed C. papaya cv. Shahi also showed the maximum number of plant regeneration compared to that of C. papaya cv. Ranchi. PMID:24066284

  2. Regulatory focus and burnout in nurses: The mediating effect of perception of transformational leadership.

    PubMed

    Shi, Rui; Zhang, Shilei; Xu, Hang; Liu, Xufeng; Miao, Danmin

    2015-12-01

    This correlation study investigated the relationship between nurses' regulatory focus and burnout, as mediated by their perceptions of transformational leadership, using a cross-sectional research design with anonymous questionnaires. In July-August 2012, data were collected from 378 nurses from three hospitals in Shaanxi Province, China, using self-report questionnaires for measuring the nurses' regulatory focus, their level of burnout and their perception of whether the leadership of their supervisor was transformational. Structural equation modelling and bootstrapping procedures were used to identify the mediating effect of their perceptions of transformational leadership. The results supported our hypothesized model. The type of regulatory focus emerged as a significant predictor of burnout. Having a perception of transformational leadership partially mediated the relationship between regulatory focus and burnout. Having a promotion focus reduced burnout when the participants perceived transformational leadership, whereas having a prevention focus exhibited the opposite pattern. The mediating effect of the perception of transformational leadership suggests that a promotion focus may help diminish burnout, directly and indirectly. Nurse managers must be aware of the role of a regulatory focus and cultivate promotion focus in their followers.

  3. Agrobacterium tumefaciens-Mediated Transformation of the Lichen Fungus, Umbilicaria muehlenbergii

    PubMed Central

    Wang, Hai-Ying; Kim, Jung A.; Yu, Nan-Hee; Kim, Sungbeom; Cheong, Yong Hwa; Kang, Seogchan; Lee, Yong-Hwan; Hur, Jae-Seoun

    2013-01-01

    Transformation-mediated mutagenesis in both targeted and random manners has been widely applied to decipher gene function in diverse fungi. However, a transformation system has not yet been established for lichen fungi, severely limiting our ability to study their biology and mechanism underpinning symbiosis via gene manipulation. Here, we report the first successful transformation of the lichen fungus, Umbilicaria muehlenbergii, via the use of Agrobacterium tumefaciens. We generated a total of 918 transformants employing a binary vector that carries the hygromycin B phosphotransferase gene as a selection marker and the enhanced green fluorescent protein gene for labeling transformants. Randomly selected transformants appeared mitotically stable, based on their maintenance of hygromycin B resistance after five generations of growth without selection. Genomic Southern blot showed that 88% of 784 transformants contained a single T-DNA insert in their genome. A number of putative mutants affected in colony color, size, and/or morphology were found among these transformants, supporting the utility of Agrobacterium tumefaciens-mediated transformation (ATMT) for random insertional mutagenesis of U. muehlenbergii. This ATMT approach potentially offers a systematic gene functional study with genome sequences of U. muehlenbergii that is currently underway. PMID:24386304

  4. Genomic regions responsible for amenability to Agrobacterium-mediated transformation in barley

    PubMed Central

    Hisano, Hiroshi; Sato, Kazuhiro

    2016-01-01

    Different plant cultivars of the same genus and species can exhibit vastly different genetic transformation efficiencies. However, the genetic factors underlying these differences in transformation rate remain largely unknown. In barley, ‘Golden Promise’ is one of a few cultivars reliable for Agrobacterium-mediated transformation. By contrast, cultivar ‘Haruna Nijo’ is recalcitrant to genetic transformation. We identified genomic regions of barley important for successful transformation with Agrobacterium, utilizing the ‘Haruna Nijo’ × ‘Golden Promise’ F2 generation and genotyping by 124 genome-wide SNP markers. We observed significant segregation distortions of these markers from the expected 1:2:1 ratio toward the ‘Golden Promise’-type in regions of chromosomes 2H and 3H, indicating that the alleles of ‘Golden Promise’ in these regions might contribute to transformation efficiency. The same regions, which we termed Transformation Amenability (TFA) regions, were also conserved in transgenic F2 plants generated from a ‘Morex’ × ‘Golden Promise’ cross. The genomic regions identified herein likely include necessary factors for Agrobacterium-mediated transformation in barley. The potential to introduce these loci into any haplotype of barley opens the door to increasing the efficiency of transformation for target alleles into any haplotype of barley by the TFA-based methods proposed in this report. PMID:27874056

  5. A new selection method for pepper transformation: callus-mediated shoot formation.

    PubMed

    Lee, Y H; Kim, H S; Kim, J Y; Jung, M; Park, Y S; Lee, J S; Choi, S H; Her, N H; Lee, J H; Hyung, N I; Lee, C H; Yang, S G; Harn, C H

    2004-08-01

    We used two genes, TMV-CP and PPI1 (pepper-PMMV interaction 1 transcription factor), to transform commercially important chili pepper (Capsicum annuum) inbred lines (P915, P409) by means of Agrobacterium co-culture. Eighteen independently transformed T0 plants were obtained. The most critical point in the pepper transformation protocol was the selection of shoots growing on calli--referred to as callus-mediated shoot formation (indirect shooting)--because shoots not grown from the callus (direct shooting from the wounded surface) developed into non-transformants. Selection of the correct right callus type also proved to be an important requirement for obtaining transformed peppers. Six different types of callus developed during the selection process. Shoots regenerated from two of these types, while one type regenerated significantly more shoots than the other types, suggesting that the capacity for shoot formation is callus type-specific. Although the transformation rate was low, transformation via callus-mediated shoot formation proved to be reproducible and was confirmed by Southern and Northern blot analyses. Based on the experimental data, we have succeeded in developing a new protocol for the selection and transformation of pepper and expect that it will be used in the future for pepper transformation.

  6. Efficient Agrobacterium-mediated transformation of the liverwort Marchantia polymorpha using regenerating thalli.

    PubMed

    Kubota, Akane; Ishizaki, Kimitsune; Hosaka, Masashi; Kohchi, Takayuki

    2013-01-01

    The thallus, the gametophyte body of the liverwort Marchantia polymorpha, develops clonal progenies called gemmae that are useful in the isolation and propagation of isogenic plants. Developmental timing is critical to Agrobacterium-mediated transformation, and high transformation efficiency has been achieved only with sporelings. Here we report an Agrobacterium-mediated transformation system for M. polymorpha using regenerating thalli. Thallus regeneration was induced by cutting the mature thallus across the apical-basal axis and incubating the basal portion of the thallus for 3 d. Regenerating thalli were infected with Agrobacterium carrying binary vector that contained a selection marker, the hygromycin phosphotransferase gene, and hygromycin-resistant transformants were obtained with an efficiency of over 60%. Southern blot analysis verified random integration of 1 to 4 copies of the T-DNA into the M. polymorpha genome. This Agrobacterium-mediated transformation system for M. polymorpha should provide opportunities to perform genetic transformation without preparing spores and to generate a sufficient number of transformants with isogenic background.

  7. Metal hybrid nanoparticles for catalytic organic and photochemical transformations.

    PubMed

    Song, Hyunjoon

    2015-03-17

    In order to understand heterogeneous catalytic reactions, model catalysts such as a single crystalline surface have been widely studied for many decades. However, catalytic systems that actually advance the reactions are three-dimensional and commonly have multiple components including active metal nanoparticles and metal oxide supports. On the other hand, as nanochemistry has rapidly been developed and been applied to various fields, many researchers have begun to discuss the impact of nanochemistry on heterogeneous catalysis. Metal hybrid nanoparticles bearing multiple components are structurally very close to the actual catalysts, and their uniform and controllable morphology is suitable for investigating the relationship between the structure and the catalytic properties in detail. In this Account, we introduce four typical structures of metal hybrid nanoparticles that can be used to conduct catalytic organic and photochemical reactions. Metal@silica (or metal oxide) yolk-shell nanoparticles, in which metal cores exist in internal voids surrounded by thin silica (or metal oxide) shells, exhibited extremely high thermal and chemical stability due to the geometrical protection of the silica layers against the metal cores. The morphology of the metal cores and the pore density of the hollow shells were precisely adjusted to optimize the reaction activity and diffusion rates of the reactants. Metal@metal oxide core-shell nanoparticles and inverted structures, where the cores supported the shells serving an active surface, exhibited high activity with no diffusion barriers for the reactants and products. These nanostructures were used as effective catalysts for various organic and gas-phase reactions, including hydrogen transfer, Suzuki coupling, and steam methane reforming. In contrast to the yolk- and core-shell structures, an asymmetric arrangement of distinct domains generated acentric dumbbells and tipped rods. A large domain of each component added multiple

  8. Contamination of the transformer oil of power transformers and shunting reactors by metal-containing colloidal particles

    SciTech Connect

    L'vov, S. Yu.; Komarov, V. B.; Bondareva, V. N.; Seliverstov, A. F.; Lyut'ko, E. O.; L'vov, Yu. N.; Ershov, B. G.

    2011-05-15

    The results of a measurement of the contamination of the oil in 66 transformers by metal-containing colloidal particles, formed as a result of the interaction of the oil with the structural materials (the copper of the windings, the iron of the tank and core etc.), and also the results of measurements of the optical turbidity of the oil in 136 transformers when they were examined at the Power Engineering Research and Development Center Company are presented. Methods of determining the concentration of copper and iron in transformer oil are considered. The limiting values of the optical turbidity factors, the copper and iron content are determined. These can serve as a basis for taking decisions on whether to replace the silica gel of the filters for continuously purifying the oil of power transformers and the shunting reactors in addition to the standardized oil contamination factors, namely, the dielectric loss tangent and the acidity number of the oil.

  9. Agrobacterium-mediated transformation of apricot (Prunus armeniaca L.) leaf explants.

    PubMed

    Petri, César; Wang, Hong; Alburquerque, Nuria; Faize, Mohamed; Burgos, Lorenzo

    2008-08-01

    A protocol for Agrobacterium-mediated stable transformation for scored, whole leaf explants of the apricot (Prunus armeniaca) cultivar Helena was developed. Regenerated shoots were selected using a two-step increased concentrations of paromomycin sulphate. Different factors affecting survival of transformed buds, including possible toxicity of green fluorescent protein (GFP) and time of exposure to high cytokine concentration in the regeneration medium, were examined. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines was 5.6%, when optimal conditions for bud survival were provided. Southern blot analysis on four randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene. This is the first time that stable transformation of an apricot cultivar is reported and constitutes also one of the few reports on the transformation of Prunus cultivars.

  10. Bcl-2 is a critical mediator of intestinal transformation

    PubMed Central

    van der Heijden, Maartje; Zimberlin, Cheryl D.; Nicholson, Anna M.; Colak, Selcuk; Kemp, Richard; Meijer, Sybren L.; Medema, Jan Paul; Greten, Florian R.; Jansen, Marnix; Winton, Douglas J.; Vermeulen, Louis

    2016-01-01

    Intestinal tumour formation is generally thought to occur following mutational events in the stem cell pool. However, active NF-κB signalling additionally facilitates malignant transformation of differentiated cells. We hypothesized that genes shared between NF-κB and intestinal stem cell (ISCs) signatures might identify common pathways that are required for malignant growth. Here, we find that the NF-κB target Bcl-2, an anti-apoptotic gene, is specifically expressed in ISCs in both mice and humans. Bcl-2 is dispensable in homeostasis and, although involved in protecting ISCs from radiation-induced damage, it is non-essential in tissue regeneration. Bcl-2 is upregulated in adenomas, and its loss or inhibition impairs outgrowth of oncogenic clones, because Bcl-2 alleviates apoptotic priming in epithelial cells following Apc loss. Furthermore, Bcl-2 expression in differentiated epithelial cells renders these cells amenable to clonogenic outgrowth. Collectively, our results indicate that Bcl-2 is required for efficient intestinal transformation following Apc-loss and constitutes a potential chemoprevention target. PMID:26956214

  11. Microstructural transformations in stress relieved type 316 stainless steel weld metal

    SciTech Connect

    Slattery, G.F.; O'Riordan, P.

    1980-02-01

    The heat treatment of Type 316L weld metal containing small amounts of ferrite results in the transformation of the ferrite to chi or ..cap alpha.. phases after aging for only 3 hr at 850/sup 0/C. The chi-phase transformation is associated with a transitional M/sub 23/C/sub 6/ carbide, whereas the sigma-phase transforms directly from the ferrite.

  12. Recent advances in chiral imino-containing ligands for metal-catalyzed asymmetric transformations.

    PubMed

    Chen, Xu; Lu, Zhan

    2017-03-21

    In this review, the recent applications of a variety of chiral imino-containing ligands classified by different types of metal-catalyzed asymmetric reactions are summarized. The progress made in this area would encourage us to design and synthesize more novel chiral imino-containing ligands, and explore their applications in metal-catalyzed asymmetric transformations.

  13. Estimation of heavy metal transformations in municipal solid waste.

    PubMed

    Flyhammar, P

    1997-05-30

    The behaviour of heavy metals bound to municipal solid waste (MSW) and exposed to 2 decades of anaerobic waste stabilization processes have been estimated. Heavy metal solid forms in a waste degradation residue have been compared with a reconstructed waste similar to that initially disposed of in 1973. The initial waste was composed of a mixture of shredded MSW (95% dry wt.) and anaerobic sewage sludge (5% dry wt.). A sequential chemical extraction method has been used to fractionate the heavy metals into five categories of available and reactive solid forms. The results imply that these forms can be ascribed to approximately 30% of the total content of the heavy metals in the degraded waste and the portion of heavy metals bound to oxidizable solid forms seems to be higher in the degraded than the fresh MSW. The bulk of the remaining heavy metals are assumed to be less available and bound into resistant lattice structures, such as metal and polymer items. A comparison between fractionation patterns of the waste in this study and of a few sediments collected from different environments imply similarities between the fresh MSW and an oxic sediment from one site and the sewage sludge and anoxic sediments from another site. Fractionation patterns of the degraded waste are found to be quite similar to those of the anoxic sediments, except for Pb, Ni and Cd which are more similar to fresh MSW.

  14. Expression of Foreign Genes Demonstrates the Effectiveness of Pollen-Mediated Transformation in Zea mays

    PubMed Central

    Yang, Liyan; Cui, Guimei; Wang, Yixue; Hao, Yaoshan; Du, Jianzhong; Zhang, Hongmei; Wang, Changbiao; Zhang, Huanhuan; Wu, Shu-Biao; Sun, Yi

    2017-01-01

    Plant genetic transformation has arguably been the core of plant improvement in recent decades. Efforts have been made to develop in planta transformation systems due to the limitations present in the tissue-culture-based methods. Herein, we report an improved in planta transformation system, and provide the evidence of reporter gene expression in pollen tube, embryos and stable transgenicity of the plants following pollen-mediated plant transformation with optimized sonication treatment of pollen. The results showed that the aeration at 4°C treatment of pollen grains in sucrose prior to sonication significantly improved the pollen viability leading to improved kernel set and transformation efficiency. Scanning electron microscopy observation revealed that the removal of operculum covering pollen pore by ultrasonication might be one of the reasons for the pollen grains to become competent for transformation. Evidences have shown that the eGfp gene was expressed in the pollen tube and embryos, and the Cry1Ac gene was detected in the subsequent T1 and T2 progenies, suggesting the successful transfer of the foreign genes to the recipient plants. The Southern blot analysis of Cry1Ac gene in T2 progenies and PCR-identified Apr gene segregation in T2 seedlings confirmed the stable inheritance of the transgene. The outcome illustrated that the pollen-mediated genetic transformation system can be widely applied in the plant improvement programs with apparent advantages over tissue-culture-based transformation methods. PMID:28377783

  15. Agrobacterium-mediated transformation of Euphorbia tirucalli callus.

    PubMed

    Uchida, Hidenobu; Yamashita, Hirofumi; Anai, Toyoaki; Muranaka, Toshiya; Ohyama, Kanji

    2010-01-01

    In order to establish a basis for transformation technology in the petroleum plant Euphorbia tirucalli, the callus of the plant was infected with Agrobacterium, washed with distilled water, sterilized with distilled water containing 100 mg/l of carbenicillin, selected on solidified B5 medium containing 13 mg/l of G418 and 100 mg/l of carbenicillin, and then on solidified B5 medium containing 25 mg/l of G418 and 100 mg/l of carbenicillin for the transgenic calli, and then the callus lines were subcultured successively on solidified B5 medium containing 50 mg/l of G418. We performed PCR analysis of sterilized G418-resistant callus line DNA and concluded that the gene introduced was integrated into the callus genome.

  16. Agrobacterium rhizogenes mediated-transformation of Asimina triloba L. cuttings.

    PubMed

    Ayala-Silva, Tomas; Beyl, Caula A; Dortch, Greg

    2007-01-01

    The current research compared rooting of pawpaw, with softwood cuttings from mature stands, cuttings from the terminal 3-4 inches of seedlings and cuttings taken from shoots of rooted cuttings from seedlings. Four treatments were used: Agrobacterium rhizogenes strain MSU-1 (A4 wild type), A. rhizogenes strain MT232 (TR105 mutant), indole-3-butyric acid at 20,000 mg L(-1) and control. Only the A. rhizogenes treatments induced rooting and only of the seedling cuttings. No statistical differences in rooting percentage were found among the bacterial strains. While the MSU-1 strain produced more roots, these were shorter and thinner than those produced by MT232 strain. No roots resulted from any other treatment. Roots obtained from infected seedlings were analyzed for transformation using polymerase chain reaction targeting the rolB and rolC vir genes. Roots confirmed as transgenic were 100% for both rolB and rolC.

  17. Transformational leadership, empowerment, and job satisfaction: the mediating role of employee empowerment.

    PubMed

    Choi, Sang Long; Goh, Chin Fei; Adam, Muhammad Badrull Hisyam; Tan, Owee Kowang

    2016-12-01

    Recent studies have revealed that nursing staff turnover remains a major problem in emerging economies. In particular, nursing staff turnover in Malaysia remains high due to a lack of job satisfaction. Despite a shortage of healthcare staff, the Malaysian government plans to create 181 000 new healthcare jobs by 2020 through the Economic Transformation Programme (ETP). This study investigated the causal relationships among perceived transformational leadership, empowerment, and job satisfaction among nurses and medical assistants in two selected large private and public hospitals in Malaysia. This study also explored the mediating effect of empowerment between transformational leadership and job satisfaction. This study used a survey to collect data from 200 nursing staff, i.e., nurses and medical assistants, employed by a large private hospital and a public hospital in Malaysia. Respondents were asked to answer 5-point Likert scale questions regarding transformational leadership, employee empowerment, and job satisfaction. Partial least squares-structural equation modeling (PLS-SEM) was used to analyze the measurement models and to estimate parameters in a path model. Statistical analysis was performed to examine whether empowerment mediated the relationship between transformational leadership and job satisfaction. This analysis showed that empowerment mediated the effect of transformational leadership on the job satisfaction in nursing staff. Employee empowerment not only is indispensable for enhancing job satisfaction but also mediates the relationship between transformational leadership and job satisfaction among nursing staff. The results of this research contribute to the literature on job satisfaction in healthcare industries by enhancing the understanding of the influences of empowerment and transformational leadership on job satisfaction among nursing staff. This study offers important policy insight for healthcare managers who seek to increase job

  18. Agrobacterium tumefaciens-mediated transformation of Valsa mali: an efficient tool for random insertion mutagenesis.

    PubMed

    Wang, Caixia; Guan, Xiangnan; Wang, Hanyan; Li, Guifang; Dong, Xiangli; Wang, Guoping; Li, Baohua

    2013-01-01

    Valsa mali is a causal agent of apple and pear trees canker disease, which is a destructive disease that causes serious economic losses in eastern Asia, especially in China. The lack of an efficient transformation system for Valsa mali retards its investigation, which poses difficulties to control the disease. In this research, a transformation system for this pathogen was established for the first time using A. tumefaciens-mediated transformation (ATMT), with the optimal transformation conditions as follows: 10(6)/mL conidia suspension, cocultivation temperature 22°C, cocultivation time 72 hours, and 200  μ M acetosyringone (AS) in the inductive medium. The average transformation efficiency was 1015.00 ± 37.35 transformants per 10(6) recipient conidia. Thirty transformants were randomly selected for further confirmation and the results showed the presence of T-DNA in all hygromycin B resistant transformants and also revealed random and single gene integration with genetic stability. Compared with wild-type strain, those transformants exhibited various differences in morphology, conidia production, and conidia germination ability. In addition, pathogenicity assays revealed that 14 transformants had mitigated pathogenicity, while one had enhanced infection ability. The results suggest that ATMT of V. mali is a useful tool to gain novel insight into this economically important pathogen at molecular levels.

  19. Agrobacterium tumefaciens-mediated genetic transformation of the entomopathogenic fungus Beauveria bassiana.

    PubMed

    dos Reis, Maria Cecília; Pelegrinelli Fungaro, Maria Helena; Delgado Duarte, Rubens Tadeu; Furlaneto, Luciana; Furlaneto, Marcia Cristina

    2004-08-01

    Agrobacterium tumefaciens-mediated transformation (agro-transformation) was successfully applied to the entomogenous fungus Beauveria bassiana. Conidia of B. bassiana were transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selective trait, under the control of a heterologous fungal promoter and the Aspergillus nidulans trpC terminator. The efficiency of transformation was up to 28 and 96 transformants per 10(4) and 10(5) target conidia, respectively, using three distinct vectors. High mitotic stability of the transformants (80-100%) was demonstrated after five successive transfers on a nonselective medium. Abortive transformants were observed for all the hph(r) vectors used. Putative transformants were analysed for the presence of the hph gene by PCR and Southern analysis. The latter analysis revealed the integration of two or more copies of the hph gene in the genome. The agro-transformation method was found to be effective for the isolation of B. bassiana hygromycin resistant transformants and may represent a useful tool for insertional mutagenesis studies in this fungus.

  20. Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses.

    PubMed

    Abello, Javier; Kelemu, Segenet; García, Celsa

    2008-03-01

    Acremonium implicatum is a seed-transmitted endophytic fungus that forms symbiotic associations with the economically significant tropical forage grasses, Brachiaria species. To take advantage of the endophyte's plant protective properties, we developed an efficient Agrobacterium-mediated transformation system for Acremonium implicatum, using green fluorescent protein (GFP) expression and vector pSK1019 (trpC promoter) or pCAMBIA1300 (CaMV35S promoter). We found that transformation efficiency doubled for both mycelial and conidial transformation as the co-cultivation period for Agrobacterium tumefaciens and Acremonium implicatum was increased from 48 to 72h. Significantly, optimal results were obtained for either mycelial or conidial transformation with Agrobacterium tumefaciens strain AGL-1 and vector pSK1019 under the control of the trpC promoter. However, mycelial transformation consistently generated a significantly higher number of transformants than did conidial transformation. The mitotic stability of the transferred DNA was confirmed by growing ten transformants in liquid and agar media for six generations. In all cases, resistance to the selection pressure (hygromycin B) was maintained. Fluorescence emission was retained by the transformants and also expressed in Brachiaria tissues from plants inoculated with GFP-transformed A. implicatum. This technology will help in the transfer and expression of agronomically important genes in host plants.

  1. Engineered metal based nanomaterials in aqueous environments: Interactions, transformations and implications

    NASA Astrophysics Data System (ADS)

    Mudunkotuwa, Imali Ama

    Nanoscience and nanotechnology offer potential routes towards addressing critical issues such as clean and sustainable energy, environmental protection and human health. Specifically, metal and metal oxide nanomaterials are found in a wide range of applications and therefore hold a greater potential of possible release into the environment or for the human to be exposed. Understanding the aqueous phase behavior of metal and metal oxide nanomaterials is a key factor in the safe design of these materials because their interactions with living systems are always mediated through the aqueous phase. Broadly the transformations in the aqueous phase can be classified as dissolution, aggregation and adsorption which are dependent and linked processes to one another. The complexity of these processes at the liquid-solid interface has therefore been one of the grand challenges that has persisted since the beginning of nanotechnology. Although classical models provide guidance for understanding dissolution and aggregation of nanoparticles in water, there are many uncertainties associated with the recent findings. This is often due to a lack of fundamental knowledge of the surface structure and surface energetics for very small particles. Therefore currently the environmental health and safety studies related to nanomaterials are more focused on understanding the surface chemistry that governs the overall processes in the liquid-solid interfacial region at the molecular level. The metal based nanomaterials focused on in this dissertation include TiO2, ZnO, Cu and CuO. These are among the most heavily used in a number of applications ranging from uses in the construction industry to cosmetic formulation. Therefore they are produced in large scale and have been detected in the environment. There is debate within the scientific community related to their safety as a result of the lack of understanding on the surface interactions that arise from the detailed nature of the surfaces

  2. Efficient Transformation of Oil Palm Protoplasts by PEG-Mediated Transfection and DNA Microinjection

    PubMed Central

    Masani, Mat Yunus Abdul; Noll, Gundula A.; Parveez, Ghulam Kadir Ahmad; Sambanthamurthi, Ravigadevi; Prüfer, Dirk

    2014-01-01

    Background Genetic engineering remains a major challenge in oil palm (Elaeis guineensis) because particle bombardment and Agrobacterium-mediated transformation are laborious and/or inefficient in this species, often producing chimeric plants and escapes. Protoplasts are beneficial as a starting material for genetic engineering because they are totipotent, and chimeras are avoided by regenerating transgenic plants from single cells. Novel approaches for the transformation of oil palm protoplasts could therefore offer a new and efficient strategy for the development of transgenic oil palm plants. Methodology/Principal Findings We recently achieved the regeneration of healthy and fertile oil palms from protoplasts. Therefore, we focused on the development of a reliable PEG-mediated transformation protocol for oil palm protoplasts by establishing and validating optimal heat shock conditions, concentrations of DNA, PEG and magnesium chloride, and the transfection procedure. We also investigated the transformation of oil palm protoplasts by DNA microinjection and successfully regenerated transgenic microcalli expressing green fluorescent protein as a visible marker to determine the efficiency of transformation. Conclusions/Significance We have established the first successful protocols for the transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. These novel protocols allow the rapid and efficient generation of non-chimeric transgenic callus and represent a significant milestone in the use of protoplasts as a starting material for the development of genetically-engineered oil palm plants. PMID:24821306

  3. Efficient transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection.

    PubMed

    Masani, Mat Yunus Abdul; Noll, Gundula A; Parveez, Ghulam Kadir Ahmad; Sambanthamurthi, Ravigadevi; Prüfer, Dirk

    2014-01-01

    Genetic engineering remains a major challenge in oil palm (Elaeis guineensis) because particle bombardment and Agrobacterium-mediated transformation are laborious and/or inefficient in this species, often producing chimeric plants and escapes. Protoplasts are beneficial as a starting material for genetic engineering because they are totipotent, and chimeras are avoided by regenerating transgenic plants from single cells. Novel approaches for the transformation of oil palm protoplasts could therefore offer a new and efficient strategy for the development of transgenic oil palm plants. We recently achieved the regeneration of healthy and fertile oil palms from protoplasts. Therefore, we focused on the development of a reliable PEG-mediated transformation protocol for oil palm protoplasts by establishing and validating optimal heat shock conditions, concentrations of DNA, PEG and magnesium chloride, and the transfection procedure. We also investigated the transformation of oil palm protoplasts by DNA microinjection and successfully regenerated transgenic microcalli expressing green fluorescent protein as a visible marker to determine the efficiency of transformation. We have established the first successful protocols for the transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. These novel protocols allow the rapid and efficient generation of non-chimeric transgenic callus and represent a significant milestone in the use of protoplasts as a starting material for the development of genetically-engineered oil palm plants.

  4. Neoplastic transformation of cells by soluble but not particulate forms of metals used in orthopaedic implants.

    PubMed

    Doran, A; Law, F C; Allen, M J; Rushton, N

    1998-01-01

    Recent developments in cell culture techniques have made it possible to study the cellular mechanisms involved in carcinogenesis and to apply these methods as screening tools in vitro. This study investigated and compared the ability of the metals most commonly used in orthopedic implants to induce toxicity and neoplastic transformation in the C3H10T1/2 mouse fibroblast cell line. Eight metals (cobalt, chromium, nickel, iron, molybdenum, aluminium, vanadium and titanium) and their alloys (stainless steel, cobalt-chrome alloy and titanium alloy) were tested, both as soluble salts and as solid particles. There were marked differences between the various metals in terms of both toxicity and transforming ability. Significant increases in the incidence of cell transformation were seen with soluble forms of cobalt, chromium, nickel and molybdenum but not with iron, aluminium, vanadium or titanium. For most of the metals. transforming ability was directly related to toxicity, although this correlation did not hold for either molybdenum or vanadium. The physical form of the metal was critically important in determining its effects, and transformation occurred only with soluble metal salts.

  5. Metal-mediated formation of fibrillar ABri amyloid.

    PubMed

    Khan, Ayesha; Ashcroft, Alison E; Korchazhkina, Olga V; Exley, Christopher

    2004-12-01

    British amyloid (ABri) peptide is precipitated as amyloid fibrils in pathological lesions which are characteristic of familial British dementia. Unlike for other amyloidogenic peptides which have been implicated in neurodegenerative disease, for example, Abeta in Alzheimer's disease and alpha synuclein in Parkinson's disease, nothing is yet known as to whether metals mediate the formation of ABri amyloid fibrils. We show herein that a concentration of ABri, which had not previously been shown to spontaneously form amyloid, formed fibrils when incubated for 12 months at 37 degrees C. The additional presence of Al(III), in particular, or Fe(III) increased significantly both the number and the size of the fibrillar amyloid deposits which were very similar in appearance to amyloid described in hippocampal plaques in familial British dementia. Co-incubation of ABri with either Zn(II) or Cu(II) precipitated the peptide but did not result in the formation of amyloid fibrils.

  6. Metal-mediated coupling of a coordinated isocyanide and indazoles.

    PubMed

    Kinzhalov, Mikhail A; Boyarskiy, Vadim P; Luzyanin, Konstantin V; Dolgushin, Fedor M; Kukushkin, Vadim Yu

    2013-08-07

    A reaction between equimolar amounts of cis-[PdCl2(CNCy)2] (1) and indazole (HInd, 2) or 5-methylindazole (HInd(Me), 3) proceeded in refluxing CHCl3 for ca. 6 h affording the aminocarbene species cis-[PdCl2{C(Ind)=N(H)Cy}(CNCy)] (4) or cis-[PdCl2{C(Ind(Me))=N(H)Cy}(CNCy)] (5) in good (72-83%) isolated yields. Complexes 4 and 5 were characterized by elemental analyses (C, H, N), HR ESI(+)-MS, IR, and 1D ((1)H, (13)C{(1)H}) and 2D ((1)H,(1)H-COSY, (1)H,(13)C-HMQC/(1)H,(13)C-HSQC, (1)H,(13)C-HMBC) NMR spectroscopies, and complex 4 as well by X-ray diffraction. The observed coupling represents the first example of metal-mediated integration between any isocyanide and any aromatic heterocyclic system having an HN center.

  7. Genes That Mediate Arsenic and Heavy Metal Detoxification in Plants

    SciTech Connect

    Lee, David A.; Gong, Ji-Ming; Schroeder, Julian I.

    2003-03-26

    To gain insight into the mechanisms of arsenic tolerance in plants, we developed a genetic screen to isolate Arabidopsis thaliana mutants with altered tolerance to arsenic. We report here on the isolation of ars1, a novel mutant with significantly increased tolerance to arsenate. ars1 accumulates similar levels of arsenic as wild type plants, but ars1 tolerance does not appear to be phytochelatin or glutathione dependent. ars1 plants do have a higher rate of phosphate uptake than wild type plants and plants grown with an excess of phosphate show increased tolerance to arsenate. Traditional models of arsenate tolerance in plants are based on the suppression of phosphate uptake pathways and, consequently, the reduced uptake of arsenate. Our data suggest that arsenate tolerance in ars1 is due to a new mechanism mediated by increased phosphate uptake in ars1. Results exploring increased metal tolerance through engineered phytochelatin expression will also be discussed.

  8. Protein-Mediated Transformation of Lipid Vesicles into Tubular Networks

    PubMed Central

    Simunovic, Mijo; Mim, Carsten; Marlovits, Thomas C.; Resch, Guenter; Unger, Vinzenz M.; Voth, Gregory A.

    2013-01-01

    Key cellular processes are frequently accompanied by protein-facilitated shape changes in the plasma membrane. N-BAR-domain protein modules generate curvature by means of complex interactions with the membrane surface. The way they assemble and the mechanism by which they operate are largely dependent on their binding density. Although the mechanism at lower densities has recently begun to emerge, how membrane scaffolds form at high densities remains unclear. By combining electron microscopy and multiscale simulations, we show that N-BAR proteins at high densities can transform a lipid vesicle into a 3D tubular network. We show that this process is a consequence of excess adhesive energy combined with the local stiffening of the membrane, which occurs in a narrow range of mechanical properties of both the membrane and the protein. We show that lipid diffusion is significantly reduced by protein binding at this density regime and even more in areas of high Gaussian curvature, indicating a potential effect on molecular transport in cells. Finally, we reveal that the breaking of the bilayer topology is accompanied by the nematic arrangement of the protein on the surface, a structural motif that likely drives the formation of reticular structures in living cells. PMID:23931319

  9. Transgenic sugar beet tolerant to imidazolinone obtained by Agrobacterium-mediated transformation.

    PubMed

    Kishchenko, E M; Komarnitskii, I K; Kuchuk, N V

    2011-01-01

    Sugar beet is highly sensitive to imidazolinone herbicides thus rotational restrictions exist. In order to develop imidazolinone tolerant sugar beets als gene from Arabidopsis thaliana encoding acetolactate synthase with S653N mutation was used for genetic transformation. Transgenic sugar beet plants were obtained by Agrobacterium-mediated transformation of aseptic seedlings using vacuum-infiltration. The efficiency of genetic transformation was 5.8%. RT-PCR analysis of obtained plants revealed accumulation of specific als transcript. The resistance to imidazolinone was proved for developed transgenic sugar beet plants in vitro and in greenhouse conditions after spraying with imazethapyr (Pursuit, BASF).

  10. Plant Cell Division Analyzed by Transient Agrobacterium-Mediated Transformation of Tobacco BY-2 Cells.

    PubMed

    Buschmann, Henrik

    2016-01-01

    The continuing analysis of plant cell division will require additional protein localization studies. This is greatly aided by GFP-technology, but plant transformation and the maintenance of transgenic lines can present a significant technical bottleneck. In this chapter I describe a method for the Agrobacterium-mediated genetic transformation of tobacco BY-2 cells. The method allows for the microscopic analysis of fluorescence-tagged proteins in dividing cells in within 2 days after starting a coculture. This transient transformation procedure requires only standard laboratory equipment. It is hoped that this rapid method would aid researchers conducting live-cell localization studies in plant mitosis and cytokinesis.

  11. Plant-mediated transformation of perchlorate into chloride

    SciTech Connect

    Nzengung, V.A.; Wang, C.; Harvey, G.

    1999-05-01

    The decontamination of perchlorate-contaminated water by woody plants was investigated in sand and hydroponic bioreactors. Willow trees were found to be the most favorable woody plants with phraetophytic characteristics in comparative screen tests with eastern cottonwoods and Eucalyptus cineria. Willows decontaminated aqueous solutions dosed with 10--100 mg/.L of perchlorate to below the method detection limit of 2 {micro}g/L. Two phytoprocesses were identified as important in the remediation of perchlorate-contaminated water: (1) uptake and phytodegradation of perchlorate in the tree branches and leaves and (2) rhizodegradation. Exposure of rooted willow trees to perchlorate-dosed media stimulated rhizodegradation. Homogeneous degradation studies using media from the root zone of dosed willow trees confirmed that rhizosphere-associated microorganisms mediated the degradation of perchlorate to chloride. Experiments conducted with varying ranges of nitrate concentrations clearly indicated that high nitrate concentrations interfered with rhizodegradation of perchlorate. This study provides evidence that the efficacy of phytoremediation of perchlorate-contaminated environments may depend on the concentration of competing terminal electron acceptors, such as nitrate, and the nitrogen source of the nutrient solution., Since perchlorate does not volatilize from water readily, a perchlorate remediation scheme may involve an intensively cultivated plantation of trees with phraetophytic characteristics and irrigation with the contaminated water.

  12. Transient plant transformation mediated by Agrobacterium tumefaciens: Principles, methods and applications.

    PubMed

    Krenek, Pavel; Samajova, Olga; Luptovciak, Ivan; Doskocilova, Anna; Komis, George; Samaj, Jozef

    2015-11-01

    Agrobacterium tumefaciens is widely used as a versatile tool for development of stably transformed model plants and crops. However, the development of Agrobacterium based transient plant transformation methods attracted substantial attention in recent years. Transient transformation methods offer several applications advancing stable transformations such as rapid and scalable recombinant protein production and in planta functional genomics studies. Herein, we highlight Agrobacterium and plant genetics factors affecting transfer of T-DNA from Agrobacterium into the plant cell nucleus and subsequent transient transgene expression. We also review recent methods concerning Agrobacterium mediated transient transformation of model plants and crops and outline key physical, physiological and genetic factors leading to their successful establishment. Of interest are especially Agrobacterium based reverse genetics studies in economically important crops relying on use of RNA interference (RNAi) or virus-induced gene silencing (VIGS) technology. The applications of Agrobacterium based transient plant transformation technology in biotech industry are presented in thorough detail. These involve production of recombinant proteins (plantibodies, vaccines and therapeutics) and effectoromics-assisted breeding of late blight resistance in potato. In addition, we also discuss biotechnological potential of recombinant GFP technology and present own examples of successful Agrobacterium mediated transient plant transformations. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. A simple and efficient Agrobacterium-mediated procedure for transformation of tomato.

    PubMed

    Sharma, Manoj K; Solanke, Amolkumar U; Jani, Dewal; Singh, Yogendra; Sharma, Arun K

    2009-09-01

    We describe a highly efficient and reproducible Agrobacterium-mediated transformation protocol applicable to several varieties of tomato (Solanum lycopersicum, earlier known as Lycopersicum esculentum). Conditions such as co-cultivation period, bacterial concentration, concentration of benzyl amino purine (BAP), zeatin and indole acetic acid (IAA) were optimized. Co-cultivation of explants with a bacterial concentration of 108 cells/ml for three days on 2 mg/l BAP, followed by regeneration on a medium containing 1 mg/ml zeatin resulted in a transformation frequency of 41.4%. Transformation of tomato plants was confirmed by Southern blot analysis and beta-glucuronidase (GUS) assay. The protocol developed showed very high efficiency of transformation for tomato varieties Pusa Ruby, Arka Vikas and Sioux. The optimized transformation procedure is simple, efficient and does not require tobacco, Petunia, tomato suspension feeder layer or acetosyringone.

  14. Agrobacterium-mediated transformation in Alpinia galanga (Linn.) Willd. for enhanced acetoxychavicol acetate production.

    PubMed

    Rao, Kiranmayee; Chodisetti, Bhuvaneswari; Mangamoori, Lakshmi Narasu; Giri, Archana

    2012-09-01

    Agrobacterium-mediated transformations ensure elevated amounts of secondary metabolite accumulation with genetic and biosynthetic stability. In the present study, Alpinia galanga rich in bioactive compounds was genetically transformed using different strains of Agrobacterium rhizogenes viz. LBA 9402, A(4), 532, 2364 and PRTGus. Even though a higher growth rate was obtained with the LBA 9402 strain, maximum acetoxychavicol acetate accumulation (ACA) was seen in the PRTGus transformant. PRTGus root line has shown 10.1 fold higher ACA content in comparison to the control roots. The lowest ACA production was shown by the A(4) transformant (4.9 fold). The quantification of ACA in the transformed roots was carried out by using HPLC, which was found to be in the order of PRTGus > LBA 9402 > 2364 > 532 > A(4). The fast growth rate of hairy roots, genetic stability and their ability to synthesize more than one metabolite offer a promising system for the production of valuable secondary metabolites.

  15. A high-efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.).

    PubMed

    Ozawa, Kenjirou

    2012-01-01

    Agrobacterium-mediated transformation of rice has been routinely performed according to the protocol reported by Hiei et al. (Plant J. 6:271-282, 1994). However, several elite japonica and many indica varieties cannot be efficiently transformed by Agrobacterium system. Also a large number of transformants are required to generate T-DNA insertion and FOX libraries as well as gene-targeting studies. To overcome these challenges, we established a high-efficiency transformation system in rice by cocultivating rice calli with Agrobacterium on filter papers moistened with enriched liquid media instead of using solid media (Ozawa, Plant Sci. 176:522-527, 2009; Ozawa and Takaiwa, Plant Sci. 179:333-337, 2010). In this system, the transformation efficiency of the calli is almost 100% in many varieties.

  16. Setaria viridis floral-dip: A simple and rapid Agrobacterium-mediated transformation method.

    PubMed

    Martins, Polyana Kelly; Nakayama, Thiago Jonas; Ribeiro, Ana Paula; Cunha, Bárbara Andrade Dias Brito da; Nepomuceno, Alexandre Lima; Harmon, Frank G; Kobayashi, Adilson Kenji; Molinari, Hugo Bruno Correa

    2015-06-01

    Setaria viridis was recently described as a new monocotyledonous model species for C4 photosynthesis research and genetic transformation. It has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model plant. We report an alternative method of S. viridis transformation using floral dip to circumvent the necessity of tissue culture phase for transgenic plant regeneration. S. viridis spikes at boot stage were selected to be immersed in Agrobacterium suspension. T1 seeds could be identified in 1.5-2 months after floral dipping. We demonstrated through molecular analysis and RFP expression that seeds and resulting plants from dipped inflorescences were transformed. Our results suggest the feasibility of S. viridis floral dip transformation as a time-saving and cost-effective compared with traditional methods. To our knowledge, this is the first report using floral dip in S. viridis as an Agrobacterium-mediated transformation method.

  17. Agrobacterium tumefaciens-mediated transformation of blueberry (Vaccinium corymbosum L.).

    PubMed

    Song, Guo-Qing; Sink, K C

    2004-12-01

    Transient expression studies using blueberry leaf explants and monitored by beta-glucuronidase (GUS) assays indicated Agrobacterium tumefaciens strain EHA105 was more effective than LBA4404 or GV3101; and the use of acetosyringone (AS) at 100 microM for inoculation and 6 days co-cultivation was optimum compared to 2, 4, 8, 10 or 12 days. Subsequently, explants of the cultivars Aurora, Bluecrop, Brigitta, and Legacy were inoculated with strain EHA105 containing the binary vector pBISN1 with the neomycin phosphotransferase gene (nptII) and an intron-interrupted GUS gene directed by the chimeric super promoter (Aocs)3AmasPmas. Co-cultivation was for 6 days on modified woody plant medium (WPM) plus 100 microM AS. Explants were then placed on modified WPM supplemented with 1.0 mg l(-1) thidiazuron, 0.5 mg l(-1) alpha-naphthaleneacetic, 10 mg l(-1) kanamycin (Km), and 250 mg l(-1) cefotaxime. Selection for Km-resistant shoots was carried out in the dark for 2 weeks followed by culture in the light at 30 microE m(-2) s(-1) at 25 degrees C. After 12 weeks, selected shoots that were both Km resistant and GUS positive were obtained from 15.3% of the inoculated leaf explants of cultivar Aurora. Sixty-eight independent clones derived from such shoots all tested positive by the polymerase chain reaction using a nptII primer. Eight of eight among these 68 clones tested positive by Southern hybridization using a gusA gene derived probe. The transformation protocol also yielded Km-resistant, GUS-positive shoots that were also PCR positive at frequencies of 5.0% for Bluecrop, 10.0% for Brigitta and 5.6% for Legacy.

  18. Use of REMI and Agrobacterium-mediated transformation to identify pathogenicity mutants of the biocontrol fungus, Coniothyrium minitans.

    PubMed

    Rogers, Christopher W; Challen, Michael P; Green, Jonathan R; Whipps, John M

    2004-12-15

    Restriction enzyme mediated integration (REMI) and Agrobacterium-mediated transformation (ATMT) were used to transform protoplasts or germinated conidia of the mycoparasite Coniothyrium minitans to hygromycin resistance. Using REMI, up to 32 transformants mug DNA(-1) were obtained, while 37.8 transformants 5 x 10(5) germlings(-1) were obtained using ATMT. Single-copy integrations occurred in 8% and 40% of REMI and ATMT transformants, respectively. A novel microtitre plate-based test was developed to expedite screening of 4000 REMI and ATMT C. minitans transformants. Nine pathogenicity mutants that displayed reduced or no pathogenicity on sclerotia of Sclerotinia sclerotiorum were identified.

  19. Transformational Leadership and Knowledge Sharing: Mediating Roles of Employee's Empowerment, Commitment, and Citizenship Behaviors

    ERIC Educational Resources Information Center

    Han, Seung Hyun; Seo, Gaeun; Yoon, Seung Won; Yoon, Dong-Yeol

    2016-01-01

    Purpose: The purpose of this paper is to empirically examine the fundamental process through which transformational leaders play a significant role in employees' knowledge sharing by investigating mediating roles of individual affects, particularly psychological empowerment, organizational commitment and organizational citizenship behavior (OCB).…

  20. Transformational Leadership and Knowledge Sharing: Mediating Roles of Employee's Empowerment, Commitment, and Citizenship Behaviors

    ERIC Educational Resources Information Center

    Han, Seung Hyun; Seo, Gaeun; Yoon, Seung Won; Yoon, Dong-Yeol

    2016-01-01

    Purpose: The purpose of this paper is to empirically examine the fundamental process through which transformational leaders play a significant role in employees' knowledge sharing by investigating mediating roles of individual affects, particularly psychological empowerment, organizational commitment and organizational citizenship behavior (OCB).…

  1. Optimized conditions for biolistic-mediated transformation of Lilium longilforum 'Nellie White'

    USDA-ARS?s Scientific Manuscript database

    A variety of tissues were used for biolistic-mediated transformation of Lilum longiflorum 'Nellie White'. Transgenic plants were not recovered from five-month-old, non-embryogenic callus or suspension cells that had been bombarded with pDM327 that contains the bar-uidA fusion gene under control the ...

  2. Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation.

    PubMed

    Shcherbak, N; Kishchenko, O; Sakhno, L; Komarnytsky, I; Kuchuk, M

    2013-01-01

    Lox sites of the Cre/lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon bar gene expression in plants obtained via Agrobacterium-mediated and biolistic transformation methods. Lox-mediated expression of bar gene, recognized by resistance of transgenic plants to PPT, occurred only in plants obtained via Agrobacterium-mediated transformation. RT-PCR analysis confirms that PPT-resistant phenotype of transgenic plants obtained via Agrobacterium-mediated transformation was caused by activation of bar gene. The plasmid with promoterless gus gene together with the lox site adjacent to the RB was constructed and transferred to Nicotiana tabacum as well. Transgenic plants exhibited GUS activity and expression of gus gene was detected in plant leaves. Expression of bar gene from the vectors containing lox site near RB allowed recovery of numerous PPT-resistant transformants of such important crops as Beta vulgaris, Brassica napus, Lactuca sativa and Solanum tuberosum. Our results demonstrate that the lox site sequence adjacent to the RB can be used to control bar gene expression in transgenic plants.

  3. Agrobacterium-mediated transformation of the ectomycorrhizal symbiont Laccaria bicolor S238N.

    PubMed

    Kemppainen, Minna; Circosta, Ariana; Tagu, Denis; Martin, Francis; Pardo, Alejandro G

    2005-12-01

    The development of an efficient transformation system is required to alter the expression of symbiosis-regulated genes and to develop insertional mutagenesis in the ectomycorrhizal basidiomycete Laccaria bicolor S238N. Vegetative mycelium of this fungus was transformed by Agrobacterium tumefaciens-mediated gene transfer. The selection marker was the hygromycin resistance gene of Escherichia coli (hph) under the control of the gpd promoter from Agaricus bisporus and the CaMV 35S terminator as part of the T-DNA. PCR amplification of hph and Southern blot analyses showed that the genome of the hygromycin-resistant transformants contained the cassette. The latter proved mostly single copy and random integration of part of the transgene into the fungal genome. A. tumefaciens-mediated gene transfer should facilitate future development of insertional mutagenesis, targeted gene disruption and RNA interference technology in L. bicolor.

  4. Transformational leadership, intrinsic motivation, and trust: a moderated-mediated model of workplace safety.

    PubMed

    Conchie, Stacey M

    2013-04-01

    Two studies examine the role of motivation and trust in the relationship between safety-specific transformational leadership and employees' safety behavior. Study 1 tested the prediction that intrinsic and identified regulation motivations mediate the relationship between safety-specific transformational leadership and employees' safety behaviors. Study 2 further explored this relationship by testing the prediction that the mediating role of intrinsic motivation is dependent on employees' level of trust in their leader. Survey data from the U.K. construction industry supported both predictions. However, the mediating role of intrinsic motivation was found only for challenge safety citizenship behaviors (i.e., voice) and not for affiliative safety citizenship behaviors (i.e., helping). These findings suggest that employees' intrinsic motivation is important to the effectiveness of leaders' efforts to promote some but not all forms of safety behavior.

  5. 25-KVA Amorphous Metal-Core Transformer Developmental Test Report

    DTIC Science & Technology

    1989-08-01

    after the DROP TEST. Tests were also performed in the Dev. Lab . Results S/N P217061 S/N P265882 S/N P265885 1.Elect.* Factry Test Lab Test Factry Test...10 RESULTS AND CONCLUSIONS ........ ..................... .. 11 Phase I .......... ........................... . 11 Phase II...characteorist ics of tleo trins formers cilan1ged as a result , of the tests. * Pass/Fail Criteria: Thin transformers had to pass tile commercial

  6. An efficient regeneration protocol for Agrobacterium-mediated transformation of melon (Cucumis melo L.).

    PubMed

    Zhang, H J; Gao, P; Wang, X Z; Luan, F S

    2014-01-08

    An efficient selection and plant regeneration protocol for Agrobacterium-mediated transformation, using cotyledon node zone-stem connection region of melon, has been developed. The new Agrobacterium-mediated transformation methodology, independent of organ culture, used the entire germinated seed as explants. The transformation system was maximized to maintain the integrity of melon itself, thus avoiding the limitations of traditional tissue culture methods. The transformation was carried out under a non-sterile environment. The incorporation of a selectable marker (neomycin phosphotransferase II) into the genome of transgenic plants was confirmed by PCR and Southern blot analyses. The transformation frequency based on the PCR was 13%. Transgenic melon plants were usually detected by PCR in less than 1 month after Agrobacterium inoculation, and seeds could be harvested in 3 months. The growth characteristics and morphology of the transgenic plants were identical to the untransformed wild-type plants. This method would be beneficial for facilitating the characteristics of gene functions and for boosting the manipulation of melon transformation for commercial purposes.

  7. [Development of genetic transformation system of Valsa mali of apple mediated by PEG].

    PubMed

    Gao, Jing; Li, Yanbo; Ke, Xiwang; Kang, Zhensheng; Huang, Lili

    2011-09-01

    The genetic transformation of Valsa mali var. mali was developed by PEG-mediated protoplasts transformation. It was transformed by PEG-induced fusion of protoplasts. The plasmid pBIG2RHPH2-GFP-GUS carrying hph gene was used and Valsa mali var. mali 03-8 isolate was used as the host strain. At 50 mg/mL driselase + 10 mg/mL lysing enzymes concentration, the mycelium of Valsa mali var. mali cultured in YEPD medium for 48 h was hydrolyzed in 10 mL enzymes liquid /0.5 g wet mycelium for 2 h. The protoplast yield was 4 x 10(7) CFU/mg. The transformation efficiency was 44 per g DNA. Analysis of the transformants by PCR and Southern blotting showed that the selectable marker gene hph was integrated effectively into the genome of Valsa mali var. mali. After 5 subculturing on PDA, 87.5% transformants could grow. This stability test of transformants suggested that the foreign gene hph was stable in heredity. This transformation system is a valuable and important tool for the further study of the pathogenic gene of Valsa mali.

  8. Hydrated lime for metals immobilization and explosives transformation: Treatability study.

    PubMed

    Martin, W Andy; Larson, S L; Nestler, C C; Fabian, G; O'Connor, G; Felt, D R

    2012-05-15

    Fragmentation grenades contain Composition B (RDX and TNT) within a steel shell casing. There is the potential for off-site migration of high explosives and metals from hand grenade training ranges by transport in surface water and subsurface transport in leachate. This treatability study used bench-scale columns and mesocosm-scale laboratory lysimeters to investigate the potential of hydrated lime as a soil amendment for in situ remediation of explosives and metals stabilization in hand grenade range soils. Compared to the unamended soil there was a 26-92% reduction of RDX in the leachate and runoff water from the lime treated soils and a 66-83% reduction of zinc in the leachate and runoff water samples; where the hand grenade range metals of concern were zinc, iron, and manganese. The amended soil was maintained at the target pH of greater than 10.5 for optimum explosives decomposition. The treatability study indicated a high potential of success for scale-up to an in situ field study.

  9. Fourier Transform Microwave Spectroscopy of Alkali Metal Hydrosulfides: Detection of Ksh

    NASA Astrophysics Data System (ADS)

    Sheridan, P. M.; Binns, M. K. L.; Young, J. P.; Bucchino, M. P.; Ziurys, L. M.

    2012-06-01

    Fourier transform microwave (FTMW) techniques have been used to record pure rotational spectra of potassium hydrosulfide and its deuterium isotopologue in their ground electronic states. This study represents the first gas phase spectroscopic observation of KSH. FTMW spectra of NaSH were also recorded. The metal hydrosulfides were produced by discharge assisted laser ablation of the solid alkali metal in the presence of hydrogen sulfide or deuterated hydrogen sulfide. Rotational transitions in the 5-20 GHz range were measured and hyperfine splittings due to the alkali metals and deuterium were resolved. Rotational as well as metal and deuterium quadrupole coupling constants have been determined from the data. The hyperfine parameters will be interpreted in terms of metal-ligand bonding character. Geometric parameters of the alkali metal hydrosulfides will be compared.

  10. Efficient Agrobacterium tumefaciens-mediated transformation and regeneration of garlic (Allium sativum) immature leaf tissue.

    PubMed

    Kenel, Fernand; Eady, Colin; Brinch, Sheree

    2010-03-01

    Transgenic garlic (Allium sativum) plants have been recovered directly from immature leaf material by selective culture following Agrobacterium-mediated transformation. This method involved the use of a binary vector containing the mgfp-ER reporter gene and hpt selectable marker, and followed a similar protocol developed previously for the transformation of immature onion embryos. The choice of tissue and post-transformation selection procedure resulted in a large increase in recovery of transgenic plants compared with previously confirmed allium transformation protocols. The presence of transgenes in the genome of the plants was confirmed using Southern analysis. This improvement in frequency and the use of clonal commercial "Printanor" germplasm now makes possible the integration of useful agronomic and quality traits into this crop.

  11. An Efficient PEG/CaCl₂-Mediated Transformation Approach for the Medicinal Fungus Wolfiporia cocos.

    PubMed

    Sun, Qiao; Wei, Wei; Zhao, Juan; Song, Jia; Peng, Fang; Zhang, Shaopeng; Zheng, Yonglian; Chen, Ping; Zhu, Wenjun

    2015-09-01

    Sclerotia of Wolfiporia cocos are of medicinal and culinary value. The genes and molecular mechanisms involved in W. cocos sclerotial formation are poorly investigated because of the lack of a suitable and reproducible transformation system for W. cocos. In this study, a PEG/ CaCl₂-mediated genetic transformation system for W. cocos was developed. The promoter Pgpd from Ganoderma lucidum effectively drove expression of the hygromycin B phosphotransferase gene in W. cocos, and approximately 30 transformants were obtained per 10 μg DNA when the protoplast suspension density was 10(6) protoplasts/ml. However, no transformants were obtained under the regulation of the PtrpC promoter from Aspergillus nidulans.

  12. Agrobacterium-mediated transformation (AMT) of Trichoderma reesei as an efficient tool for random insertional mutagenesis.

    PubMed

    Zhong, Yao Hua; Wang, Xiao Li; Wang, Tian Hong; Jiang, Qiao

    2007-01-01

    Filamentous fungus Trichoderma reesei QM9414 was successfully transformed with Agrobacterium tumefaciens AGL-1 for random integration of transforming DNA (T-DNA). Co-cultivation of T. reesei conidia or protoplasts with A. tumefaciens in the presence of acetosyringone resulted in the formation of hygromycin B-resistant fungal colonies with high transformation frequency. Nine randomly selected resistant clones were proved to be stable through mitotic cell division. The integration of the hph gene into T. reesei genome was determined by PCR and dot blot analysis. Transgenic T. reesei strains were analyzed using TAIL-PCR for their T-DNA contents. The results showed that T-DNA inserts occurred evidently by fusing DNA at T-DNA borders via random recombination, which suggests that Agrobacterium-mediated transformation is a potentially powerful tool towards tagged mutagenesis and gene transfer technology for T. reesei.

  13. The migration and transformation behaviors of heavy metals during the hydrothermal treatment of sewage sludge.

    PubMed

    Huang, Hua-Jun; Yuan, Xing-Zhong

    2016-01-01

    Various hydrothermal treatment methods, including hydrothermal carbonization, liquefaction and sub/super-critical water gasification, have been applied to the disposal of sewage sludge for producing bio-materials or bio-fuels. It has become a research hotspot whether the heavy metals contained in sewage sludge can be well treated/stabilized after the hydrothermal treatments. This review firstly summarized the methods of assessing heavy metals' contamination level/risk and then discussed the migration and transformation behaviors of heavy metals from the following aspects: the effect of reaction temperature, the effect of additives (catalysts and other biomass), the effect of the type of solvent and the effect of reaction time. This review can provide an important reference for the further study of the migration and transformation behaviors of heavy metals during the hydrothermal treatment of sewage sludge. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Transformation of metals speciation in a combined landfill leachate treatment.

    PubMed

    Wu, Yanyu; Zhou, Shaoqi; Chen, Dongyu; Zhao, Rong; Li, Huosheng; Lin, Yiming

    2011-04-01

    Landfill leachate was treated by a combined sequential batch reactor (SBR), coagulation, Fenton oxidation and biological aerated filter (BAF) technology. The metals in treatment process were fractionated into three fractions: particulate and colloidal (size charge filtration), free ion/labile (cation exchange) and non-labile fractions. Fifty percent to 66% Cu, Ni, Zn, Mn, Pb and Cd were present as particulate/colloidal matter in raw leachate, whereas Cr was present 94.9% as non-labile complexes. The free ion/labile fractions of Ni, Zn, Mg, Mn, Pb and Cd increased significantly after treatment except Cr. Fifty-nine percent to 100% of Al was present mainly as particulate/colloidal matter >0.45 μm and the remaining portions were predicted as non-labile complexes except in coagulation effluent. The speciation of Fe varied significantly in various individual processes. Visual MINTEQ simulation showed that 95-100% colloidal species for Cu, Cd and Pb were present as metal-humic complexes even with the lower dissolved organic carbon. Optimum agreements for the free ion/labile species were within acidic solution, whereas under-estimated in alkaline effluents. Overestimated particulate/colloidal fraction consisted with the hypothesis that a portion of colloids in fraction <0.45 μm were considered as dissolved.

  15. Catalytic and photocatalytic transformations on metal nanoparticles with targeted geometric and plasmonic properties.

    PubMed

    Linic, Suljo; Christopher, Phillip; Xin, Hongliang; Marimuthu, Andiappan

    2013-08-20

    Heterogeneous catalysis by metals was among the first enabling technologies that extensively relied on nanoscience. The early intersections of catalysis and nanoscience focused on the synthesis of catalytic materials with high surface to volume ratio. These synthesis strategies mainly involved the impregnation of metal salts on high surface area supports. This would usually yield quasi-spherical nanoparticles capped by low-energy surface facets, typically with closely packed metal atoms. These high density areas often function as the catalytically active surface sites. Unfortunately, strategies to control the functioning surface facet (i.e., the geometry of active sites that performs catalytic turnover) are rare and represent a significant challenge in our ability to fine-tune and optimize the reactive surfaces. Through recent developments in colloidal chemistry, chemists have been able to synthesize metallic nanoparticles of both targeted size and desired shape. This has opened new possibilities for the design of heterogeneous catalytic materials, since metal nanoparticles of different shapes are terminated with different surface facets. By controlling the surface facet exposed to reactants, we can start affecting the chemical transformations taking place on the metal particles and changing the outcome of catalytic processes. Controlling the size and shape of metal nanoparticles also allows us to control the optical properties of these materials. For example, noble metals nanoparticles (Au, Ag, Cu) interact with UV-vis light through an excitation of localized surface plasmon resonance (LSPR), which is highly sensitive to the size and shape of the nanostructures. This excitation is accompanied by the creation of short-lived energetic electrons on the surface of the nanostructure. We showed recently that these energetic electrons could drive photocatalytic transformations on these nanostructures. The photocatalytic, electron-driven processes on metal nanoparticles

  16. Identification of virulence genes in the corn pathogen Colletotrichum graminicola by Agrobacterium tumefaciens-mediated transformation.

    PubMed

    Münch, Steffen; Ludwig, Nancy; Floss, Daniela S; Sugui, Janyce A; Koszucka, Anna M; Voll, Lars M; Sonnewald, Uwe; Deising, Holger B

    2011-01-01

    A previously developed Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for the plant pathogenic fungus Colletotrichum graminicola led to high rates of tandem integration of the whole Ti-plasmid, and was therefore considered to be unsuitable for the identification of pathogenicity and virulence genes by insertional mutagenesis in this pathogen. We used a modified ATMT protocol with acetosyringone present only during the co-cultivation of C. graminicola and A. tumefaciens. Analysis of 105 single-spore isolates randomly chosen from a collection of approximately 2000 transformants, indicated that almost 70% of the transformants had single T-DNA integrations. Of 500 independent transformants tested, 10 exhibited attenuated virulence in infection assays on whole plants. Microscopic analyses primarily revealed defects at different pre-penetration stages of infection-related morphogenesis. Three transformants were characterized in detail. The identification of the T-DNA integration sites was performed by amplification of genomic DNA ends after endonuclease digestion and polynucleotide tailing. In one transformant, the T-DNA had integrated into the 5'-flank of a gene with similarity to allantoicase genes of other Ascomycota. In the second and third transformants, the T-DNA had integrated into an open reading frame (ORF) and into the 5'-flank of an ORF. In both cases, the ORFs have unknown function.

  17. Agrobacterium-mediated genetic transformation of the desiccation tolerant resurrection plant Ramonda myconi (L.) Rchb.

    PubMed

    Tóth, Sándor; Kiss, Csaba; Scott, Peter; Kovács, Gabriella; Sorvari, Seppo; Toldi, Ottó

    2006-05-01

    In this paper we describe the first procedure for Agrobacterium tumefaciens-mediated genetic transformation of the desiccation tolerant plant Ramonda myconi (L.) Rchb. Previously, we reported the establishment of a reliable and effective tissue culture system based on the integrated optimisation of antioxidant and growth regulator composition and the stabilisation of the pH of the culture media by means of a potassium phosphate buffer. This efficient plant regeneration via callus phase provided a basis for the optimisation of the genetic transformation in R. myconi. For gene delivery, both a standard (method A) and a modified protocol (method B) have been applied. Since the latter has previously resulted in successful transformation of another resurrection plant, Craterostigma plantagineum, an identical protocol was utilized in transformation of R. myconi, as this method may prove general for dicotyledonous resurrection plants. On this basis, physical and biochemical key variables in transformation were evaluated such as mechanical microwounding of plant explants and in vitro preinduction of vir genes. While the physical enhancement of bacterial penetration was proved to be essential for successful genetic transformation of R. myconi, an additional two-fold increase in the transformation frequency was obtained when the above physical and biochemical treatments were applied in combination. All R0 and R1 transgenic plants were fertile, and no morphological abnormalities were observed on the whole-plant level.

  18. Stable Agrobacterium-mediated transformation of maritime pine based on kanamycin selection.

    PubMed

    Alvarez, José M; Ordás, Ricardo J

    2013-01-01

    An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β -glucuronidase (uidA) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL(-1) kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density (OD(600 nm)) of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth.

  19. Stable Agrobacterium-Mediated Transformation of Maritime Pine Based on Kanamycin Selection

    PubMed Central

    Alvarez, José M.; Ordás, Ricardo J.

    2013-01-01

    An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β-glucuronidase (uidA) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL−1 kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density (OD600 nm) of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth. PMID:24376383

  20. Agrobacterium-Mediated Stable Genetic Transformation of Populus angustifolia and Populus balsamifera

    PubMed Central

    Maheshwari, Priti; Kovalchuk, Igor

    2016-01-01

    The present study demonstrates Agrobacterium tumefaciens-mediated stable genetic transformation of two species of poplar – Populus angustifolia and Populus balsamifera. The binary vector pCAMBIA-Npro-long-Luc containing the luciferase reporter gene was used to transform stem internode and axillary bud explants. Putative transformants were regenerated on selection-free medium using our previously established in vitro regeneration method. Explant type, genotype, effect of pre-culture, Agrobacterium concentration, a time period of infection and varying periods of co-culture with bacteria were tested for the transformation frequency. The highest frequency of transformation was obtained with stem internode explants pre-cultured for 2 days, infected with Agrobacterium culture at the concentration of OD600 = 0.5 for 10 min and co-cultivated with Agrobacterium for 48 h. Out of the two genotypes tested, P. balsamifera exhibited a higher transformation rate in comparison to P. angustifolia. The primary transformants that exhibited luciferase activity in a bioluminescence assay under the CCD camera when subjected to polymerase chain reaction and Southern blot analysis revealed a stable single-copy integration of luc in their genomes. The reported protocol is highly reproducible and can be applied to other species of poplar; it will also be useful for future genetic engineering of one of the most important families of woody plants for sustainable development. PMID:27014319

  1. Agrobacterium-mediated large-scale transformation of wheat (Triticum aestivum L.) using glyphosate selection.

    PubMed

    Hu, T; Metz, S; Chay, C; Zhou, H P; Biest, N; Chen, G; Cheng, M; Feng, X; Radionenko, M; Lu, F; Fry, J

    2003-06-01

    An Agrobacterium-mediated transformation system with glyphosate selection has been developed for the large-scale production of transgenic plants. The system uses 4-day precultured immature embryos as explants. A total of 30 vectors containing the 5-enol-pyruvylshikimate-3-phosphate synthase gene from Agrobacterium strain CP4 (aroA:CP4), which confers resistance to glyphosate, were introduced into wheat using this system. The aroA:CP4 gene served two roles in this study-selectable marker and gene of interest. More than 3,000 transgenic events were produced with an average transformation efficiency of 4.4%. The entire process from isolation of immature embryos to production of transgenic plantlets was 50-80 days. Transgenic events were evaluated over several generations based on genetic, agronomic and molecular criteria. Forty-six percent of the transgenic events fit a 3:1 segregation ratio. Molecular analysis confirmed that four of six lead transgenic events selected from Agrobacterium transformation contained a single insert and a single copy of the transgene. Stable expression of theAROA:CP4 gene was confirmed by ELISA through nine generations. A comparison of Agrobacterium-mediated transformation to a particle bombardment system demonstrated that the Agrobacterium system is reproducible, has a higher transformation efficiency with glyphosate selection and produces higher quality transgenic events in wheat. One of the lead events from this study, no. 33391, has been identified as a Roundup Ready wheat commercial candidate.

  2. Transformational leadership and safety performance among nurses: the mediating role of knowledge-related job characteristics.

    PubMed

    Lievens, Ilse; Vlerick, Peter

    2014-03-01

    To report the impact of transformational leadership on two dimensions of nurses' safety performance (i.e. safety compliance and safety participation) and to study the mediating role of knowledge-related job characteristics in this relationship. Safety performance refers to the behaviours that employees exhibit to adhere to safety guidelines and to promote health and safety at their workplace. Nurses' safety performance is a major challenge for healthcare settings, urging the need to identify the key determinants and psychological mechanisms that influence it. A cross-sectional survey study. The study was carried out in September 2010 in a large Belgian hospital. We used self-administered questionnaires; 152 nurses participated. The hypotheses were tested using hierarchical regression analyses. In line with our first hypothesis, the results show that transformational leadership exerted a significant positive impact on both dimensions of nurses' safety performance. This positive relation was mediated by knowledge-related job characteristics, supporting our second hypothesis. Head nurses' transformational leadership can enhance nurses' compliance with and participation in safety. Furthermore, transformational head nurses are able to influence the perception that their nurses have about the kind and amount of knowledge in their job, which can also lead to increases in both dimensions of nurses' safety performance. This study therefore demonstrates the key impact that transformational head nurses have, both directly and indirectly, on the safety performance of their nurses. © 2013 John Wiley & Sons Ltd.

  3. Parameters affecting the efficiency of Agrobacterium tumefaciens-mediated transformation of Colletotrichum graminicola.

    PubMed

    Flowers, Jennifer L; Vaillancourt, Lisa J

    2005-12-01

    We have developed an Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for the plant pathogenic fungus Colletotrichum graminicola, the cause of anthracnose leaf blight and stalk rot of corn. The ATMT results in higher transformation efficiencies than previously available polyethylene glycol-mediated protocols, and falcate spores can be used instead of protoplasts for transformation. Various experimental parameters were tested for their effects on transformation efficiencies. The parameters with the greatest influence were the A. tumefaciens strain used and the Ti-plasmid it carried, the ratio of bacterium to fungus during cocultivation, and the length of cocultivation. Southern analysis demonstrated that most transformants (80%) contained tandem integrations of plasmid sequences, and at least 36% had integrations at multiple sites in the genome. In a majority of cases (70%), the whole Ti-plasmid, and not just the T-DNA, had integrated as a series of tandem repeats. Tandem integrations, especially of the whole plasmid, make it difficult to rescue DNA from both flanks of the integrations with standard PCR-based approaches. Thus, ATMT may be unsuitable for insertional mutagenesis of C. graminicola without further modification.

  4. Metastable crystal growth of acetaminophen using solution-mediated phase transformation

    NASA Astrophysics Data System (ADS)

    Mori, Yoichiro; Maruyama, Mihoko; Takahashi, Yoshinori; Yoshikawa, Hiroshi Y.; Okada, Shino; Adachi, Hiroaki; Sugiyama, Shigeru; Takano, Kazufumi; Murakami, Satoshi; Matsumura, Hiroyoshi; Inoue, Tsuyoshi; Yoshimura, Masashi; Mori, Yusuke

    2017-01-01

    We report a new method of obtaining the metastable phase form II crystals of acetaminophen. Solution-mediated phase transformation (SMPT) from trihydrate into form II is utilized to obtain form II crystals. SMPT is triggered by seeding form II crystals into a saturated solution including trihydrate crystals, which are less stable than form II crystals. Form II seed crystals gradually grew at the expense of the dissolving trihydrate crystals, and finally, all the trihydrate crystals in solution were transformed into form II crystals in about 4 h. Thus, we conclude that SMPT is effective for the production of form II crystals.

  5. Acinetobacter baumannii Coordinates Urea Metabolism with Metal Import To Resist Host-Mediated Metal Limitation.

    PubMed

    Juttukonda, Lillian J; Chazin, Walter J; Skaar, Eric P

    2016-09-27

    During infection, bacterial pathogens must adapt to a nutrient metal-limited environment that is imposed by the host. The innate immune protein calprotectin inhibits bacterial growth in vitro by chelating the divalent metal ions zinc (Zn(2+), Zn) and manganese (Mn(2+), Mn), but pathogenic bacteria are able to cause disease in the presence of this antimicrobial protein in vivo. One such pathogen is Acinetobacter baumannii, a Gram-negative bacterium that causes pneumonia and bloodstream infections that can be complicated by resistance to multiple antibiotics. A. baumannii inhibition by calprotectin is dependent on calprotectin Mn binding, but the mechanisms employed by A. baumannii to overcome Mn limitation have not been identified. This work demonstrates that A. baumannii coordinates transcription of an NRAMP family Mn transporter and a urea carboxylase to resist the antimicrobial activities of calprotectin. This NRAMP family transporter facilitates Mn accumulation and growth of A. baumannii in the presence of calprotectin. A. baumannii is found to utilize urea as a sole nitrogen source, and urea utilization requires the urea carboxylase encoded in an operon with the NRAMP family transporter. Moreover, urea carboxylase activity is essential for calprotectin resistance in A. baumannii Finally, evidence is provided that this system combats calprotectin in vivo, as deletion of the transporter impairs A. baumannii fitness in a mouse model of pneumonia, and this fitness defect is modulated by the presence of calprotectin. These findings reveal that A. baumannii has evolved mechanisms to subvert host-mediated metal sequestration and they uncover a connection between metal starvation and metabolic stress.

  6. Acinetobacter baumannii Coordinates Urea Metabolism with Metal Import To Resist Host-Mediated Metal Limitation

    PubMed Central

    Juttukonda, Lillian J.; Chazin, Walter J.

    2016-01-01

    ABSTRACT During infection, bacterial pathogens must adapt to a nutrient metal-limited environment that is imposed by the host. The innate immune protein calprotectin inhibits bacterial growth in vitro by chelating the divalent metal ions zinc (Zn2+, Zn) and manganese (Mn2+, Mn), but pathogenic bacteria are able to cause disease in the presence of this antimicrobial protein in vivo. One such pathogen is Acinetobacter baumannii, a Gram-negative bacterium that causes pneumonia and bloodstream infections that can be complicated by resistance to multiple antibiotics. A. baumannii inhibition by calprotectin is dependent on calprotectin Mn binding, but the mechanisms employed by A. baumannii to overcome Mn limitation have not been identified. This work demonstrates that A. baumannii coordinates transcription of an NRAMP family Mn transporter and a urea carboxylase to resist the antimicrobial activities of calprotectin. This NRAMP family transporter facilitates Mn accumulation and growth of A. baumannii in the presence of calprotectin. A. baumannii is found to utilize urea as a sole nitrogen source, and urea utilization requires the urea carboxylase encoded in an operon with the NRAMP family transporter. Moreover, urea carboxylase activity is essential for calprotectin resistance in A. baumannii. Finally, evidence is provided that this system combats calprotectin in vivo, as deletion of the transporter impairs A. baumannii fitness in a mouse model of pneumonia, and this fitness defect is modulated by the presence of calprotectin. These findings reveal that A. baumannii has evolved mechanisms to subvert host-mediated metal sequestration and they uncover a connection between metal starvation and metabolic stress. PMID:27677795

  7. A metal mixture induces transformation upon antioxidant depletion in a hepatic cell line.

    PubMed

    Sánchez-Valle, Vicente; Valverde, Mahara; Carrizale, Leticia; Mejía, Jesus; Zepeta, Nahum; Rojas, Emilio

    2013-01-01

    Metals are ubiquitous soil, air, and water pollutants. A mixture of arsenic cadmium and lead, in particular, has commonly been found in the vicinity of smelter areas. The mixture of As-Cd-Pb has been shown to be carcinogenic, and transforming potential and oxidative stress have been proposed as principal mechanisms involved in this process. The aim of this work was to explore the role of the antioxidant barrier in the establishment of cell transformation upon chronic exposure to a metal mixture containing 2 μM NaAsO(2), 2 μM. CdCl(2), and 5 μM Pb(C(2)H(3)O(2))(2)∙3H(2)O in WRL-68 cells-a non-transformed human hepatic cell line. In this study, we used a WRL-68 cell model of human embryonic hepatic origin treated with antioxidant inhibitors (L-Buthionine-sulfoxamine and aminotriazole) to test the role of the antioxidant barrier in the establishment of cell transformation upon chronic exposure to a metal mixture of As-Cd-Pb (2 μM NaAsO(2), 2 μM CdCl(2) and 5 μM Pb(C(2)H(3)O(2))(2)∙3H(2)O). We evaluated oxidative damage markers, including reactive oxygen species, lipid peroxidation, and genotoxicity, as well as antioxidant response markers, including glutathione concentration, catalase activity, and superoxide dismutase activity, which promote morphological transformation, which can be quantified by foci formation. As expected, we found an increase in the intracellular concentration of the metals after treatment with the metal mixture. In addition, treatment with the metal mixture in addition to inhibitors resulted in a large increase in the intracellular concentration of cadmium and lead. Our results describe the generation of reactive oxygen species, cytotoxicity, genotoxicity, and oxidative damage to macromolecules that occurred exclusively in cells that were morphologically transformed upon exposure to a metal mixture and antioxidant barrier inhibition. Our results show the importance of the antioxidant barrier role in the protection of cellular integrity

  8. Agrobacterium-mediated transformation of oat (Avena sativa L.) cultivars via immature embryo and leaf explants.

    PubMed

    Gasparis, Sebastian; Bregier, Cezary; Orczyk, Waclaw; Nadolska-Orczyk, Anna

    2008-11-01

    This paper reports on the successful Agrobacterium-mediated transformation of oat, and on some factors influencing this process. In the first step of the experiments, three cultivars, two types of explant, and three combinations of strain/vectors, which were successfully used for transformation of other cereals were tested. Transgenic plants were obtained from the immature embryos of cvs. Bajka, Slawko and Akt and from leaf base explants of cv. Bajka after transformation with A. thumefaciens strain LBA4404(pTOK233). The highest transformation rate (12.3%) was obtained for immature embryos of cv. Bajka. About 79% of the selected plants proved to be transgenic; however, only 14.3% of the T(0) plants and 27.5% of the T(1) showed GUS expression. Cell competence of both types of explant differed in terms of their transformation ability and transgene expression. The next step of the study was to test the suitability for oat transformation of the pGreen binary vector combined with different selection cassettes: nptII or bar under the nos or 35S promoter. Transgenic plants were selected in combinations transformed with nos::nptII, 35S::nptII and nos::bar. The highest transformation efficiency (5.3%) was obtained for cv. Akt transformed with nos::nptII. A detailed analysis of the T(0) plants selected from a given callus line and their progeny revealed that they were the mixture of transgenic, chimeric-transgenic and non-transgenic individuals. Southern blot analysis of T(0) and T(1) showed simple integration pattern with the low copy number of the introduced transgenes.

  9. Efficient ΦC31 integrase-mediated site-specific germline transformation of Anopheles gambiae.

    PubMed

    Pondeville, Emilie; Puchot, Nicolas; Meredith, Janet M; Lynd, Amy; Vernick, Kenneth D; Lycett, Gareth J; Eggleston, Paul; Bourgouin, Catherine

    2014-07-01

    Current transgenic methodology developed for mosquitoes has not been applied widely to the major malaria vector Anopheles gambiae, which has proved more difficult to genetically manipulate than other mosquito species and dipteran insects. In this protocol, we describe ΦC31-mediated site-specific integration of transgenes into the genome of A. gambiae. The ΦC31 system has many advantages over 'classical' transposon-mediated germline transformation systems, because it allows integration of large transgenes at specific, characterized genomic locations. Starting from a general protocol, we have optimized steps from embryo collection to co-injection of transgene-containing plasmid and in vitro-produced ΦC31 integrase mRNA. We also provide tips for screening transgenic larvae. The outlined procedure provides robust transformation in A. gambiae, resulting in homozygous transgenic lines in ∼2-3 months.

  10. High Pressure Phase Transformations in Heavy Rare Earth Metals and Connections to Actinide Crystal Structures

    SciTech Connect

    Vohra, Yogesh K.; Sangala, Bagvanth Reddy; Stemshorn, Andrew K.; Hope, Kevin M.

    2008-07-01

    High-pressure studies have been performed on heavy rare earth metals Terbium (Tb) to 155 GPa and Holmium (Ho) to 134 GPa in a diamond anvil cell at room temperature. The following crystal structure sequence was observed in both metals hcp {yields} Sm-type {yields} dhcp {yields} distorted fcc (hR-24) {yields} monoclinic (C2/m) with increasing pressure. The last transformation to a low symmetry monoclinic phase is accompanied by a volume collapse of 5 % for Tb at 51 GPa and a volume collapse of 3 % for Ho at 103 GPa. This volume collapse under high pressure is reminiscent of f-shell delocalization in light rare earth metal Cerium (Ce), Praseodymium (Pr), and heavy actinide metals Americium (Am) and Curium (Cm). The orthorhombic Pnma phase that has been reported in Am and Cm after f-shell delocalization is not observed in heavy rare earth metals under high pressures. (authors)

  11. Enhanced Endosomal Escape by Light-Fueled Liquid-Metal Transformer.

    PubMed

    Lu, Yue; Lin, Yiliang; Chen, Zhaowei; Hu, Quanyin; Liu, Yang; Yu, Shuangjiang; Gao, Wei; Dickey, Michael D; Gu, Zhen

    2017-04-12

    Effective endosomal escape remains as the "holy grail" for endocytosis-based intracellular drug delivery. To date, most of the endosomal escape strategies rely on small molecules, cationic polymers, or pore-forming proteins, which are often limited by the systemic toxicity and lack of specificity. We describe here a light-fueled liquid-metal transformer for effective endosomal escape-facilitated cargo delivery via a chemical-mechanical process. The nanoscale transformer can be prepared by a simple approach of sonicating a low-toxicity liquid-metal. When coated with graphene quantum dots (GQDs), the resulting nanospheres demonstrate the ability to absorb and convert photoenergy to drive the simultaneous phase separation and morphological transformation of the inner liquid-metal core. The morphological transformation from nanospheres to hollow nanorods with a remarkable change of aspect ratio can physically disrupt the endosomal membrane to promote endosomal escape of payloads. This metal-based nanotransformer equipped with GQDs provides a new strategy for facilitating effective endosomal escape to achieve spatiotemporally controlled drug delivery with enhanced efficacy.

  12. Theory of ordering transformations in metals and minerals

    SciTech Connect

    Lindsey, T.F. . Dept. of Materials Science and Mineral Engineering Lawrence Berkeley Lab., CA )

    1991-07-01

    This dissertation presents an investigation of ordering in FCC based systems using the pair potential approximation in the ground state and mean field limits. The theoretical approach is used to explain the occurrence of observed equilibrium phases and characteristics of thermodynamic instabilities, in particular, spinodal ordering and decomposition. It is shown that the stability of non-integer domain sizes in long period superstructures such as Al{sub 3}Ti and Ag{sub 3}Mg may result from the tendency of a system to reduce the number of non-dominant ordering waves, thus producing domain sizes that have rational fraction form n/m. This conclusion is used to explain the domain size stability with respect to variations in temperature and electron concentration. The cation ordering in the precipitate phases in calcite and dolomite is analyzed by analogy with ordering in FCC based metals. The ordered phases in calcite and dolomite are shown to be consistent with pair potential minima at {l brace}100{r brace} and {l brace}1/2, 1/2, 1/2{r brace} positions in reciprocal space respectively. 32 refs., 6 figs.

  13. Quantification of Grain Boundary Mediated Plasticity Mechanisms in Nanocrystalline Metals

    NASA Astrophysics Data System (ADS)

    Panzarino, Jason F.

    Nanocrystalline metals have been a topic of great discussion over recent years due to their exceptional strengths and novel grain boundary-mediated deformation mechanisms. Their microstructures are known to evolve through dynamic processes such as grain boundary migration and grain rotation, but how the collective interaction of these mechanisms alter the microstructure on a larger scale is not completely understood. In this thesis, we present coupled atomistic modeling and experimental tasks that aim to understand how the grain structure, grain boundaries, and associated grain boundary network change during nanocrystalline plasticity. Due to the complex three-dimensional nature of these mechanisms and the limited spatial and temporal resolution of current in-situ experimental techniques, we turn to atomistic modeling to help understand the dynamics by which these mechanisms unfold. In order to provide a quantitative analysis of this behavior, we develop a tool which fully characterizes nanocrystalline microstructures in atomistic models and subsequently tracks their evolution during molecular dynamics simulations. We then use this algorithm to quantitatively track grain structure and boundary network evolution in plastically deformed nanocrystalline Al, finding that higher testing temperature and smaller average grain size results in increased evolution of grain structure with evidence of larger scale changes to the grain boundary network also taking place. This prompts us to extend our analysis technique to include full characterization of grain boundary networks and rigorous topographical feature identification. We then employ this tool on simulations of Al subject to monotonic tension, cycling loading, and simple annealing, and find that each case results in different evolution of the grain boundary network. Finally, our computational work is complemented synergistically by experimental analyses which track surface microstructure evolution during sliding wear

  14. Optimization of factors affecting Agrobacterium-mediated transformation of Micro-Tom tomatoes.

    PubMed

    Guo, M; Zhang, Y L; Meng, Z J; Jiang, J

    2012-03-16

    Micro-Tom is the smallest known variety of tomatoes. An orthogonal experimental design L(16) (4(5)) was used to optimize Agrobacterium-mediated transformation of cotyledon explants of Lycopersicon esculentum cv. Micro-Tom. Four parameters were investigated to determine their effect on transformation frequency: the concentration of bacterial suspension, time of dip in bacterial suspension, co-cultivation time, and concentration of carbenicillin. We also examined the effect of these parameters on contamination rate, necrosis rate, mortality, cut-surface browning rate, and undamaged explant rate. Both the bacterial and carbenicillin concentrations had a significant influence on the rate of infected explants. The time of co-cultivation also had a significant influence on the transformation parameters. The optimal transformation protocol consisted of an Agrobacterium suspension of 0.5 × 10(8) cells/mL (OD(600) = 0.5) and an infection time of 5 min, one day of co-cultivation and 500 mg/L carbenicillin. Under these conditions, the transformation efficiency of the shoots reached 5.1%; the mean transformation frequency was 3.9% (N = 838).

  15. Efficient sweet pepper transformation mediated by the BABY BOOM transcription factor.

    PubMed

    Heidmann, Iris; de Lange, Brenda; Lambalk, Joep; Angenent, Gerco C; Boutilier, Kim

    2011-06-01

    Pepper (Capsicum L.) is a nutritionally and economically important crop that is cultivated throughout the world as a vegetable, condiment, and food additive. Genetic transformation using Agrobacterium tumefaciens (agrobacterium) is a powerful biotechnology tool that could be used in pepper to develop community-based functional genomics resources and to introduce important agronomic traits. However, pepper is considered to be highly recalcitrant for agrobacterium-mediated transformation, and current transformation protocols are either inefficient, cumbersome or highly genotype dependent. The main bottleneck in pepper transformation is the inability to generate cells that are competent for both regeneration and transformation. Here, we report that ectopic expression of the Brassica napus BABY BOOM AP2/ERF transcription factor overcomes this bottleneck and can be used to efficiently regenerate transgenic plants from otherwise recalcitrant sweet pepper (C. annuum) varieties. Transient activation of BABY BOOM in the progeny plants induced prolific cell regeneration and was used to produce a large number of somatic embryos that could be converted readily to seedlings. The data highlight the utility of combining biotechnology and classical plant tissue culture approaches to develop an efficient transformation and regeneration system for a highly recalcitrant vegetable crop.

  16. Agrobacterium tumefaciens-mediated transformation of corn (Zea mays L.) multiple shoots

    PubMed Central

    Cao, Shi-liang; Masilamany, Pathmalojiny; Li, Wen-bin; Pauls, K. Peter

    2014-01-01

    An Agrobacterium tumefaciens-mediated corn transformation method based on multiple shoot tissue cultures was developed, which is effective with a variety of corn inbred lines and standard binary vectors. Six factors that affected the success of corn transformation were tested, including A. tumefaciens strain, corn genotype, tissue culture growth stage, medium composition, co-culture temperature and surfactant treatment. Agropine-type bacteria (EHA 101 and AGL 1) were eightfold more effective than octopine-type strain for corn multi-shoot tissues transformation. The average frequency of Glucuronidase (GUS)-positive explants obtained from 14 corn genotypes ranged from 36% to 76%. L-proline (0.7 g L−1) in the co-culture medium apparently improved the frequency of transformation. The newly initiated multi-shoot tissues were most responsive to Agrobacterium infection. A positive correlation was found between multi-shoot tissue susceptibility to Agrobacterium and the proportion of cells in G1 phase. Transformants were identified by reverse transcription Polymerase Chain Reaction (PCR) and by southern blot hybridization assays. The frequency of transformants was approximately 2% based on the number of multi-shoot explants co-cultivated with Agrobacterium. PMID:26019506

  17. Agrobacterium tumefaciens-mediated transformation of Penicillium expansum PE-12 and its application in molecular breeding.

    PubMed

    Zhang, Tian; Qi, Zhen; Wang, Yueyue; Zhang, Fangyuan; Li, Renyong; Yu, Qingsheng; Chen, Xiangbin; Wang, Huojun; Xiong, Xin; Tang, Kexuan

    2013-03-30

    Lipase produced by Penicillium expansum is widely used in laundry detergent and leather industry; however, the absence of an efficient transformation technology sets a major obstacle for further enhancement of its lipase productivity through advanced gene engineering. In this work, Agrobacterium tumefaciens-mediated transformation (ATMT) was investigated for P. expansum PE-12 transformation, using hygromycin phosphotransferase (hph) as a selectable marker gene. As a result, we revealed that the frequency of transformation surpassed 100 transformants/10(5)condida, most of the integrated T-DNA appeared as a single copy at a random position in chromosomal DNA, and all the transformants showed mitotic stability. Facilitated by this newly established method, for the first time, P. expansum PE-12 was genetically engineered to improve the lipase yield, through a homologous expression vector carrying the endogenous lipase gene (PEL) driven by the strong constitutive promoter of the glyceraldehydes-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans. The highest expression level of the engineered strain reached up to 1700 U/mL, nearly 2-fold of the original industrial strain (900 U/mL). Our reproducible ATMT system has not only revealed the great potential of homologous expression-directed genetic engineering, which is more efficient and specific compared to traditional mutagenesis, but also provided new possibilities and perspectives for any other practical applications of P. expansum-related genetic engineering in the future.

  18. Agrobacterium tumefaciens-mediated transformation of the entomopathogenic fungus Nomuraea rileyi.

    PubMed

    Shao, Changwen; Yin, Youping; Qi, Zhaoran; Li, Ren; Song, Zhangyong; Li, Yan; Wang, Zhongkang

    2015-10-01

    An Agrobacterium-mediated genetic transformation system for the entomopathogenic fungus Nomuraea rileyi was established. Three binary T-DNA vectors, pPZP-Hph, pPZP-Hph-RNAi and pPZP-Hph-DsRed2, were constructed. The trpc promoter from Aspergillus nidulans was used as the cis-regulatory element to drive the expression of hygromycin phosphotransferase (hph) gene and DsRed2, which conferred the hygromycin B (Hyg B) resistance and red fluorescence visualization, respectively. The blastospores and conidia were used as the recipients. The blastospores' transformation efficiency reached ∼20-40 transformants per 10(6) blastospores, whereas the conidia were not transformed. Based on an analysis of five generations of subcultures, PCR and Southern blotting assays, the Ptrpc-hph cassette had integrated into the genomes of all transformants, which contained single copy of the hph gene and showed mitotic stability. Abundant altered morphologic phenotypes in colonies, blastospores and hyphae formations were observed in the arbitrary insertional mutants of N. rileyi, which made it possible to study the relationships between the functions and the interrupted genes over the whole genome. The transformation protocol will promote the functional characterization of genes, and the construction of genetically engineered strains of this important entomopathogenic fungus, and potentially of other similar fungal pathogens.

  19. Transition-metal-free acid-mediated synthesis of aryl sulfides from thiols and thioethers.

    PubMed

    Wagner, Anna M; Sanford, Melanie S

    2014-03-07

    The preparation of diaryl and alkyl aryl sulfides via acid-mediated coupling of thiols and thioethers with diaryliodonium salts is reported. The scope, limitations, and mechanism of the transformation are discussed.

  20. X-ray absorption spectroscopy (XAS) of toxic metal mineral transformations by fungi.

    PubMed

    Fomina, Marina; Charnock, John; Bowen, Andrew D; Gadd, Geoffrey M

    2007-02-01

    Fungi can be highly efficient biogeochemical agents and accumulators of soluble and particulate forms of metals. This work aims to understand some of the physico-chemical mechanisms involved in toxic metal transformations focusing on the speciation of metals accumulated by fungi and mycorrhizal associations. The amorphous state or poor crystallinity of metal complexes within biomass and relatively low metal concentrations make the determination of metal speciation in biological systems a challenging problem but this can be overcome by using synchrotron-based element-specific X-ray absorption spectroscopy (XAS) techniques. In this research, we have exposed fungi and ectomycorrhizas to a variety of copper-, zinc- and lead-containing minerals. X-ray absorption spectroscopy studies revealed that oxygen ligands (phosphate, carboxylate) played a major role in toxic metal coordination within the fungal and ectomycorrhizal biomass during the accumulation of mobilized toxic metals. Coordination of toxic metals within biomass depended on the fungal species, initial mineral composition, the nitrogen source, and the physiological state/age of the fungal mycelium.

  1. Strategies to improve low copy transgenic events in Agrobacterium-mediated transformation of maize.

    PubMed

    Sivamani, Elumalai; Li, Xianggan; Nalapalli, Samson; Barron, Yoshimi; Prairie, Anna; Bradley, David; Doyle, Michele; Que, Qiudeng

    2015-12-01

    Transgenic plants containing low copy transgene insertion free of vector backbone are highly desired for many biotechnological applications. We have investigated two different strategies for increasing the percentage of low copy events in Agrobacterium-mediated transformation experiments in maize. One of the strategies is to use a binary vector with two separate T-DNAs, one T-DNA containing an intact E.coli manA gene encoding phosphomannose isomerase (PMI) as selectable marker gene cassette and another T-DNA containing an RNAi cassette of PMI sequences. By using this strategy, low copy transgenic events containing the transgenes were increased from 43 to 60 % in maize. An alternate strategy is using selectable marker gene cassettes containing regulatory or coding sequences derived from essential plant genes such as 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) or MADS box transcription factor. In this paper we demonstrate that higher percentage of low copy transgenic events can be obtained in Agrobacterium-mediated maize transformation experiments using both strategies. We propose that the above two strategies can be used independently or in combination to increase transgenic events that contain low copy transgene insertion in Agrobacterium-mediated transformation experiments.

  2. Advances in Agrobacterium tumefaciens-mediated genetic transformation of graminaceous crops.

    PubMed

    Singh, Roshan Kumar; Prasad, Manoj

    2016-05-01

    Steady increase in global population poses several challenges to plant science research, including demand for increased crop productivity, grain yield, nutritional quality and improved tolerance to different environmental factors. Transgene-based approaches are promising to address these challenges by transferring potential candidate genes to host organisms through different strategies. Agrobacterium-mediated gene transfer is one such strategy which is well known for enabling efficient gene transfer in both monocot and dicots. Due to its versatility, this technique underwent several advancements including development of improved in vitro plant regeneration system, co-cultivation and selection methods, and use of hyper-virulent strains of Agrobacterium tumefaciens harbouring super-binary vectors. The efficiency of this method has also been enhanced by the use of acetosyringone to induce the activity of vir genes, silver nitrate to reduce the Agrobacterium-induced necrosis and cysteine to avoid callus browning during co-cultivation. In the last two decades, extensive efforts have been invested towards achieving efficient Agrobacterium-mediated transformation in cereals. Though high-efficiency transformation systems have been developed for rice and maize, comparatively lesser progress has been reported in other graminaceous crops. In this context, the present review discusses the progress made in Agrobacterium-mediated transformation system in rice, maize, wheat, barley, sorghum, sugarcane, Brachypodium, millets, bioenergy and forage and turf grasses. In addition, it also provides an overview of the genes that have been recently transferred to these graminaceous crops using Agrobacterium, bottlenecks in this technique and future possibilities for crop improvement.

  3. Fabrication of mesoporous metal oxide coated-nanocarbon hybrid materials via a polyol-mediated self-assembly process

    NASA Astrophysics Data System (ADS)

    Feng, Bingmei; Wang, Huixin; Wang, Dongniu; Yu, Huilong; Chu, Yi; Fang, Hai-Tao

    2014-11-01

    After clarifying the formation mechanism of a typical metal glycolate precipitate, Ti glycolate, in a polyol-mediated synthesis using acetone as a precipitation medium, we describe a simple template-free approach based on an ethylene glycol-mediated synthesis to fabricate mesoporous metal oxide coated-nanocarbon hybrid materials including TiO2 coated-carbon nanotube (CNT), SnO2 coated-CNT, Cu2O/CuO coated-CNT and TiO2 coated-graphene sheet (GS). In the approach, metal oxide precursors, metal glycolates, were first deposited on CNTs or GSs, and subsequently transformed to the metal oxide coatings by pyrolysis or hydrolysis. By a comparison between the characterization of two TiO2-CNT hybrid materials using carboxylated CNTs and pristine CNTs without carboxyl groups, the driving force for initiating the deposition of metal glycolates on the carboxylated CNTs is confirmed to be the hydrogen bonding between the carboxyl groups and the polymer chains in metal glycolate sols. The electrochemical performances of the mesoporous TiO2 coated-carboxylated CNTs and TiO2-pristine CNT hybrid materials were investigated. The results show that the mesoporous TiO2 coated-carboxylated CNT with a uniform core-shell nanostructure exhibits substantial improvement in the rate performance in comparison with its counterpart from 0.5 C to 100 C because of its higher electronic conductivity and shorter diffusion path for the lithium ion. At the extremely high rate of 100 C, the specific capacity of TiO2 of the former reaches 85 mA h g-1, twice as high as that of the latter.After clarifying the formation mechanism of a typical metal glycolate precipitate, Ti glycolate, in a polyol-mediated synthesis using acetone as a precipitation medium, we describe a simple template-free approach based on an ethylene glycol-mediated synthesis to fabricate mesoporous metal oxide coated-nanocarbon hybrid materials including TiO2 coated-carbon nanotube (CNT), SnO2 coated-CNT, Cu2O/CuO coated-CNT and TiO2

  4. N-14 NQR using a high-Tc rf SQUID with a normal metal transformer

    NASA Astrophysics Data System (ADS)

    He, D. F.; Tachiki, M.; Itozaki, H.

    2008-01-01

    We have improved our high-Tc SQUID-based N-14 nuclear quadrupole resonance (NQR) detection system. By using a reed relay SIL5-1A72-71L (MEDER electronic) in the normal metal transformer, the isolation to the excitation field was much improved. The high-Tc rf SQUID could operate stably when the rf excitation field was over 20 mT. By optimizing the input coil of the transformer, better sensitivity of 0.5 fT Hz-1/2 was obtained at the resonant frequency of the tuned normal metal transformer. A spin-locking spin-echo (SLSE) multi-pulse sequence was also used in the system to detect the weak NQR signals produced by samples with longer spin-spin relaxation time T2, such as trinitrotoluene (TNT).

  5. Mechanistic Variants in Gas-Phase Metal-Oxide Mediated Activation of Methane at Ambient Conditions.

    PubMed

    Li, Jilai; Zhou, Shaodong; Zhang, Jun; Schlangen, Maria; Usharani, Dandamudi; Shaik, Sason; Schwarz, Helmut

    2016-09-07

    The C-H bond activation of methane mediated by a prototypical heteronuclear metal-oxide cluster, [Al2Mg2O5](•+), was investigated by using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) in conjunction with high-level quantum mechanical calculations. Experimentally, hydrogen-atom abstraction from methane by the cluster ion [Al2Mg2O5](•+) takes place at ambient conditions. As to the mechanism, according to our computational findings, both the proton-coupled electron transfer (PCET) and the conventional hydrogen-atom transfer (HAT) are feasible and compete with each other. This is in distinct contrast to the [XYO2](+) (X, Y = Mg, Al, Si) cluster oxide ions which activate methane exclusively via the PCET route (Li, J.; Zhou, S.; Zhang, J.; Schlangen, M.; Weiske, T.; Usharani, D.; Shaik, S.; Schwarz, H. J. Am. Chem. Soc. 2016, 138, 7973-7981). The electronic origins of the mechanistically rather complex reactivity scenarios of the [Al2Mg2O5](•+)/CH4 couple were elucidated. For the PCET mechanism, in which the Lewis acid-base pair [Al(+)-O(-)] of the cluster acts as the active site, a clear correlation has been established between the nature of the transition state, the corresponding barrier height, the Lewis acidity-basicity of the [M(+)-O(-)] unit, as well as the bond order of the M(+)-O(-) bond. Also addressed is the role of the spin and charge distributions of a terminal oxygen radical site in the direct HAT route. The knowledge of the factors that control the reactivity of PCET and HAT pathways not only deepens our mechanistic understanding of metal-oxide mediated C-H bond activation but may also provide guidance for the rational design of catalysts.

  6. Mitochondrial oligomers boost glycolysis in cancer stem cells to facilitate blebbishield-mediated transformation after apoptosis

    PubMed Central

    Jinesh, GG; Molina, JR; Huang, L; Laing, NM; Mills, GB; Bar-Eli, M; Kamat, AM

    2016-01-01

    Apoptosis culminates in secondary necrosis due to lack of ATP. Cancer stem cells form spheres after apoptosis by evoking the blebbishield emergency program. Hence, determining how blebbishields avoid secondary necrosis is crucial. Here we demonstrate that N-Myc and VEGFR2 control transformation from blebbishields, during which oligomers of K-Ras, p27, BAD, Bax, and Bak boost glycolysis to avoid secondary necrosis. Non-apoptotic cancer cells also utilize oligomers to boost glycolysis, which differentiates the glycolytic function of oligomers from their apoptotic action. Smac mimetic in combination with TNF-α or TRAIL but not in combination with FasL abrogates transformation from blebbishields by inducing secondary necrosis. Thus blebbishield-mediated transformation is dependent on glycolysis, and Smac mimetics represent potential candidates to abrogate the blebbishield emergency program. PMID:27551498

  7. The Mechanism of Porosity Formation During Solvent-Mediated Phase Transformations

    SciTech Connect

    Christophe Raufaste; Bjorn Jamtveit; Timm John; Paul Meakin; Dag Kristian Dysthe

    2011-05-01

    Solvent-mediated solid–solid phase transformations often result in the formation of a porous medium, which may be stable on long time scales or undergo ripening and consolidation. We have studied replacement processes in the KBr–KCl–H2O system using both in situ and ex situ experiments. The replacement of a KBr crystal by a K(Br,Cl) solid solution in the presence of an aqueous solution is facilitated by the generation of a surprisingly stable, highly anisotropic and connected pore structure that pervades the product phase. This pore structure ensures efficient solute transport from the bulk solution to the reacting KBr and K(Br,Cl) surfaces. The compositional profile of the K(Br,Cl) solid solution exhibits striking discontinuities across disc-like cavities in the product phase. Similar transformation mechanisms are probably important in controlling phase-transformation processes and rates in a variety of natural and man-made systems.

  8. Mitochondrial oligomers boost glycolysis in cancer stem cells to facilitate blebbishield-mediated transformation after apoptosis.

    PubMed

    Jinesh, G G; Molina, J R; Huang, L; Laing, N M; Mills, G B; Bar-Eli, M; Kamat, A M

    2016-01-01

    Apoptosis culminates in secondary necrosis due to lack of ATP. Cancer stem cells form spheres after apoptosis by evoking the blebbishield emergency program. Hence, determining how blebbishields avoid secondary necrosis is crucial. Here we demonstrate that N-Myc and VEGFR2 control transformation from blebbishields, during which oligomers of K-Ras, p27, BAD, Bax, and Bak boost glycolysis to avoid secondary necrosis. Non-apoptotic cancer cells also utilize oligomers to boost glycolysis, which differentiates the glycolytic function of oligomers from their apoptotic action. Smac mimetic in combination with TNF-α or TRAIL but not in combination with FasL abrogates transformation from blebbishields by inducing secondary necrosis. Thus blebbishield-mediated transformation is dependent on glycolysis, and Smac mimetics represent potential candidates to abrogate the blebbishield emergency program.

  9. Functional analysis of the white gene of Drosophila by P-factor-mediated transformation.

    PubMed

    Gehring, W J; Klemenz, R; Weber, U; Kloter, U

    1984-09-01

    A 12-kb DNA segment spanning the white (w) locus of Drosophila has been inserted into a P-transposon vector and used for P-factor-mediated germ-line transformation. Several red-eyed transformants were recovered which complement the white mutant phenotype. Analysis of the eye pigments and the interaction with the zeste mutation indicates that the w gene inserted at several new chromosomal sites is expressed normally. The tissue-specific accumulation of w transcripts, as studied by in situ hybridization to tissue sections, is the same in transformant and wild-type larvae. This indicates that all the genetic information specified by the w locus is contained within this 12-kb segment of DNA. By secondary mobilization it was shown that the w sequences have been inserted as a functional P(w) transposon which is capable of further transposition.

  10. Relationship between transformational leadership style and organizational commitment: Mediating effect of psychological empowerment

    NASA Astrophysics Data System (ADS)

    Asif, Muhammad; Ayyub, Samia; Bashir, Muhammad Khawar

    2014-12-01

    This study explores the relationship between style of transformational leadership and organizational commitment of employees with mediating role of psychological empowerment in the textile sector Punjab Pakistan. Data was collected using tools from 250 employees. The transformational leadership questionnaire, MLQ-Multifactor leadership Questionnaire [1] was used to verify the perception of the employees towards transformational leadership style in two dimensions i.e. idealized influence and inspirational motivation. The organizational commitment questionnaire designed by [2] was used to verify the affective organizational commitment. Further, psychological empowerment questionnaire was developed by [3] which was used to examine the state of psychological empowerment of textile sector employees. Pearson Correlation revealed that there exists a positive significant relationship between idealized influence and affective organizational commitment, Inspirational motivation and affective organizational commitment, affective organizational commitment and psychological empowerment. The results from the study put forward that there is a significant relationship between style of transformational leadership and organizational commitment. The mediating variable which one is suitable in the model i.e. psychological empowerment and the model is good fit as the F value is significant.

  11. Toward "metalloMOFzymes": Metal-Organic Frameworks with Single-Site Metal Catalysts for Small-Molecule Transformations.

    PubMed

    Cohen, Seth M; Zhang, Zhenjie; Boissonnault, Jake A

    2016-08-01

    Metal-organic frameworks (MOFs) are being increasingly studied as scaffolds and supports for catalysis. The solid-state structures of MOFs, combined with their high porosity, suggest that MOFs may possess advantages shared by both heterogeneous and homogeneous catalysts, with few of the shortcomings of either. Herein, efforts to create single-site catalytic metal centers appended to the organic ligand struts of MOFs will be discussed. Reactions important for advanced energy applications, such as H2 production and CO2 reduction, will be highlighted. Examining how these active sites can be introduced, their performance, and their existing limitations should provide direction for design of the next generation of MOF-based catalysts for energy-relevant, small-molecule transformations. Finally, the introduction of second-sphere interactions (e.g., hydrogen bonding via squaramide groups) as a possible route to enhancing the activity of these metal centers is reported.

  12. Nanoscale Twinning and Martensitic Transformation in Shock-Deformed BCC Metals

    SciTech Connect

    Hsiung, L L

    2005-03-22

    Shock-induced twinning and martensitic transformation in BCC-based polycrystalline metals (Ta and U-6wt%Nb) have been observed and studied using transmission electron microscopy (TEM). The length-scale of domain thickness for both twin lamella and martensite phase is found to be smaller than 100 nm. While deformation twinning of {l_brace}112{r_brace}<111>-type is found in Ta when shock-deformed at 15 GPa, both twinning and martensitic transformation are found in Ta when shock-deformed at 45 GPa. Similar phenomena of nanoscale twinning and martensitic transformation are also found in U6Nb shock-deformed at 30 GPa. Since both deformation twinning and martensitic transformation occurred along the {l_brace}211{r_brace}{sub b} planes associated with high resolved shear stresses, it is suggested that both can be regarded as alternative paths for shear transformations to occur in shock-deformed BCC metals. Heterogeneous nucleation mechanisms for shock-induced twinning and martensitic transformation are proposed and discussed.

  13. Development of an Agrobacterium-mediated stable transformation method for the sensitive plant Mimosa pudica.

    PubMed

    Mano, Hiroaki; Fujii, Tomomi; Sumikawa, Naomi; Hiwatashi, Yuji; Hasebe, Mitsuyasu

    2014-01-01

    The sensitive plant Mimosa pudica has long attracted the interest of researchers due to its spectacular leaf movements in response to touch or other external stimuli. Although various aspects of this seismonastic movement have been elucidated by histological, physiological, biochemical, and behavioral approaches, the lack of reverse genetic tools has hampered the investigation of molecular mechanisms involved in these processes. To overcome this obstacle, we developed an efficient genetic transformation method for M. pudica mediated by Agrobacterium tumefaciens (Agrobacterium). We found that the cotyledonary node explant is suitable for Agrobacterium-mediated transformation because of its high frequency of shoot formation, which was most efficiently induced on medium containing 0.5 µg/ml of a synthetic cytokinin, 6-benzylaminopurine (BAP). Transformation efficiency of cotyledonary node cells was improved from almost 0 to 30.8 positive signals arising from the intron-sGFP reporter gene by using Agrobacterium carrying a super-binary vector pSB111 and stabilizing the pH of the co-cultivation medium with 2-(N-morpholino)ethanesulfonic acid (MES) buffer. Furthermore, treatment of the explants with the detergent Silwet L-77 prior to co-cultivation led to a two-fold increase in the number of transformed shoot buds. Rooting of the regenerated shoots was efficiently induced by cultivation on irrigated vermiculite. The entire procedure for generating transgenic plants achieved a transformation frequency of 18.8%, which is comparable to frequencies obtained for other recalcitrant legumes, such as soybean (Glycine max) and pea (Pisum sativum). The transgene was stably integrated into the host genome and was inherited across generations, without affecting the seismonastic or nyctinastic movements of the plants. This transformation method thus provides an effective genetic tool for studying genes involved in M. pudica movements.

  14. Development of an Agrobacterium-Mediated Stable Transformation Method for the Sensitive Plant Mimosa pudica

    PubMed Central

    Mano, Hiroaki; Fujii, Tomomi; Sumikawa, Naomi; Hiwatashi, Yuji; Hasebe, Mitsuyasu

    2014-01-01

    The sensitive plant Mimosa pudica has long attracted the interest of researchers due to its spectacular leaf movements in response to touch or other external stimuli. Although various aspects of this seismonastic movement have been elucidated by histological, physiological, biochemical, and behavioral approaches, the lack of reverse genetic tools has hampered the investigation of molecular mechanisms involved in these processes. To overcome this obstacle, we developed an efficient genetic transformation method for M. pudica mediated by Agrobacterium tumefaciens (Agrobacterium). We found that the cotyledonary node explant is suitable for Agrobacterium-mediated transformation because of its high frequency of shoot formation, which was most efficiently induced on medium containing 0.5 µg/ml of a synthetic cytokinin, 6-benzylaminopurine (BAP). Transformation efficiency of cotyledonary node cells was improved from almost 0 to 30.8 positive signals arising from the intron-sGFP reporter gene by using Agrobacterium carrying a super-binary vector pSB111 and stabilizing the pH of the co-cultivation medium with 2-(N-morpholino)ethanesulfonic acid (MES) buffer. Furthermore, treatment of the explants with the detergent Silwet L-77 prior to co-cultivation led to a two-fold increase in the number of transformed shoot buds. Rooting of the regenerated shoots was efficiently induced by cultivation on irrigated vermiculite. The entire procedure for generating transgenic plants achieved a transformation frequency of 18.8%, which is comparable to frequencies obtained for other recalcitrant legumes, such as soybean (Glycine max) and pea (Pisum sativum). The transgene was stably integrated into the host genome and was inherited across generations, without affecting the seismonastic or nyctinastic movements of the plants. This transformation method thus provides an effective genetic tool for studying genes involved in M. pudica movements. PMID:24533121

  15. Metal-mediated DNA damage and cell death: mechanisms, detection methods, and cellular consequences.

    PubMed

    Angelé-Martínez, Carlos; Goodman, Craig; Brumaghim, Julia

    2014-08-01

    The redox activity of metal ions can lead to the formation of highly reactive species that damage DNA, producing different oxidation products and types of damage depending upon the redox potentials of the DNA bases, formation of intermediate adducts, and identity of the reactive species. Other factors are also important in determining the degree of metal-mediated DNA damage, such as localization and redox chemistry of the metal ions or complexes and lifetimes of the reactive oxygen species generated. This review examines the types of DNA damage mediated by first-row transition metals under oxidative stress conditions, with emphasis on work published in the past ten years. Similarities and differences between DNA damage mechanisms of the first-row transition metals in vitro and in E. coli and human cells are compared and their relationship to disease development are discussed. Methods to detect this metal-mediated DNA damage, including backbone breakage, base oxidation, inter- and intra-strand crosslinking, and DNA-protein crosslinking are also briefly reviewed, as well as detection methods for reactive oxygen species generated by these metal ions. Understanding the conditions that cause metal-mediated DNA damage and metal generation of reactive oxygen species in vitro and in cells is required to develop effective drugs to prevent and treat chronic disease.

  16. Hydrogen-Induced Phase Transformations: a Base for a New Sphere of the Science of Metals (an Analytical Review)

    NASA Astrophysics Data System (ADS)

    Goltsov, V. A.

    2017-03-01

    Data on the phase transformations induced in metals by hydrogen are generalized and analyzed. It is suggested to classify hydrogen-induced phase transformations on the basis of allowance for the temperature dependence of the parameters of diffusion of substitutional and interstitial (hydrogen) atoms like in the classical science of metals. A pioneer classification of hydrogen-induced phase transformations by this method is developed.

  17. Agrobacterium tumefaciens-mediated transformation of Withania somnifera (L.) Dunal: an important medicinal plant.

    PubMed

    Pandey, Vibha; Misra, Pratibha; Chaturvedi, Pankaj; Mishra, Manoj K; Trivedi, Prabodh K; Tuli, Rakesh

    2010-02-01

    This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a-half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l(-1) kanamycin. Polymerase chain reaction analysis of T(0) transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T(1) progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was found to be 1.67%.

  18. Highly efficient transformation and plant regeneration of tall fescue mediated by Agrobacterium tumefaciens.

    PubMed

    Hu, Zhang-Hua; Chen, Jin-Qing; Wu, Guan-Ting; Jin, Wei; Lang, Chun-Xiu; Huang, Rui-Zhi; Wang, Fu-Lin; Liu, Zhi-Hong; Chen, Xiao-Yun

    2005-04-01

    An efficient and reproducible system has been developed for the production of transgenic plants in tall fescue (Festuca arundinacea Schreb.) using A. tumefaciens-mediated transformation. Two-months-old suspension cultures served as excellent explants for transformation. The explants were inoculated with an A. tumefaciens strain EHA105 carrying a plasmid pDBA121 containing genes for hygromycin phosphotransferase (hpt) and phosphinotricin acetyltransferase (bar). The commercial herbicide Basta was used as a selective agent. Inclusion of acetosyringone (ACS) 20 mg/L in the co-culture medium led to an increase in transformation efficiency. The efficiency of transformation was highly dependent on the genotype, the explant, the culture medium and selective agent used. Tall fescue transformation efficiency is 2.85-10.9 plants per gram fresh weight (FW) of suspension cultures. This is much higher than the corresponding values reported before (2-5 plants). So far more than 300 transgenic plants have been obtained, the fertility of some transgenic plants had decreased. Stable integration and high expression of the transgenes were confirmed by PCR analysis and Southern blot hybridization or herbicide Basta spraying test.

  19. Integrative gene transfer in the truffle Tuber borchii by Agrobacterium tumefaciens-mediated transformation.

    PubMed

    Brenna, Andrea; Montanini, Barbara; Muggiano, Eleonora; Proietto, Marco; Filetici, Patrizia; Ottonello, Simone; Ballario, Paola

    2014-01-01

    Agrobacterium tumefaciens-mediated transformation is a powerful tool for reverse genetics and functional genomic analysis in a wide variety of plants and fungi. Tuber spp. are ecologically important and gastronomically prized fungi ("truffles") with a cryptic life cycle, a subterranean habitat and a symbiotic, but also facultative saprophytic lifestyle. The genome of a representative member of this group of fungi has recently been sequenced. However, because of their poor genetic tractability, including transformation, truffles have so far eluded in-depth functional genomic investigations. Here we report that A. tumefaciens can infect Tuber borchii mycelia, thereby conveying its transfer DNA with the production of stably integrated transformants. We constructed two new binary plasmids (pABr1 and pABr3) and tested them as improved transformation vectors using the green fluorescent protein as reporter gene and hygromycin phosphotransferase as selection marker. Transformants were stable for at least 12 months of in vitro culture propagation and, as revealed by TAIL- PCR analysis, integration sites appear to be heterogeneous, with a preference for repeat element-containing genome sites.

  20. Agrobacterium tumefaciens-mediated transformation of the causative agent of Valsa canker of apple tree Valsa mali var. mali.

    PubMed

    Hu, Yang; Dai, Qingqing; Liu, Yangyang; Yang, Zhe; Song, Na; Gao, Xiaoning; Voegele, Ralf Thomas; Kang, Zhensheng; Huang, Lili

    2014-06-01

    Valsa mali var. mali (Vmm), which is the causative agent of Valsa canker of apple tree, causes heavy damage to apple production in eastern Asia. In this article, we report Agrobacterium tumefaciens-mediated transformation (ATMT) of Vmm and expression of gfp (green fluorescent protein) in this fungus. The transformation system was optimized to a transformation efficiency of approximately 150 transformants/10(6) conidia, and a library containing over 4,000 transformants was generated. The tested transformants were mitotically stable. One hundred percent hph (hygromycin B phosphotransferase) integration into Vmm was identified by PCR and five single-copy integration of T-DNA was detected in the eighteen transformants by Southern blot. To our knowledge, this is the first report of ATMT of Vmm. Furthermore, this library has been used to identify genes involved in the virulence of the pathogen, and the transformation system may also be useful to the transformation of other species of the genus Valsa.

  1. Agrobacterium tumefaciens-mediated genetic transformation of Salix matsudana Koidz. using mature seeds.

    PubMed

    Yang, Jingli; Yi, Jaeseon; Yang, Chuanping; Li, Chenghao

    2013-06-01

    An Agrobacterium tumefaciens-mediated transformation method was developed for Salix matsudana Koidz. using mature seeds as starting material. Multiple shoots were induced directly from embryonic shoot apices of germinating seeds. Although thidiazuron, 6-benzylaminopurine and zeatin induced multiple shoot induction with high frequency, zeatin (4.5 μM) was more effective for elongation of shoots and roots. The binary vector pCAMBIA1303, which contained neomycin phosphotransferase as a selectable marker gene and β-glucuronidase as a reporter gene, was used for transformation. Factors affecting transformation efficiency were examined for optimization of the procedure. Up to 35 of 180 seeds regenerated kanamycin-resistant shoots under optimal transformation conditions as follows: seeds were precultured for 4 days, apices of embryonic shoots were removed and infected with A. tumefaciens strain LBA4404 grown to a cell density equivalent (OD600) of 0.6, and then the infected explants were cultivated at 21 °C for 4 days. Storage of seeds at -20 °C for as long as 3 years had no significant effect on the induction of kanamycin-resistant shoots. Using this method, transgenic plants were obtained within ∼5 months with a transformation frequency of 7.2%. Analysis by polymerase chain reaction (PCR) showed that 36.4-93.8% of plants from all 13 tested kanamycin-resistant lines were PCR positive. Several 'escapes' were eliminated by a second round of selection. PCR, Southern blot and reverse transcriptase-PCR analyses of selected transgenic individuals 2 years after cutting propagation confirmed the successful generation of stable transformants. Our method, which minimizes the duration of axenic culture, may provide an alternative procedure for transformation of other recalcitrant Salix species.

  2. [Genetic transformation of OSISAP1 gene to onion (Allium cepa L.) mediated by amicroprojectile bombardment].

    PubMed

    Xu, Qi-Jiang; Cui, Cheng-Ri

    2007-06-01

    Microprojectile bombardment-mediated transformation method has been developed for onion (Allium cepa L.) using embryogenic calli, induced from stem discs, as target tissue. Zinc-finger protein gene OSISAP1 (Oryza sative subspecies indica stress-associated protein gene) was introduced into the open-pollinated onion cultivar (subs.) 'HG400B'. Bombardment parameters were optimized as: the pressure is 1,100 psi, the distance is 6 cm, two times, the ratio of mass between plasmid DNA and golden particles is 1:320. An efficient microprojectile bombardment-mediated transformation system of onion (Allium cepa L.) callus has been established. The binary vector used carried the nptII gene for kanamycin resistance and the GUS reporter gene. Transgenic cultures were screened for their ability to express the GUS reporter gene and to grow in the presence of kanamycin (150 mg/L). Transient expression of GUS reporter gene was observed through histochemical staining of embryogenic callus transformed by microprojectile bombardment. The putative transgenic plants were analysed at the molecular level using PCR, southern hybridization, and RT-PCR. The results confirmed that the OSISAP1 gene was integrated as one copy into the genome of onion and expression. Transgenic plants were produced efficiently with a transformation frequency of about 10%. Test of salinity-alkali stress showed that sodium chloride and sodium bicarbonate at 200 mmol/L effectively killed non-transgenic plants within 1 week of irrigation, while the transgenic plants were completely unaffected by salinity of 400 mmol/L. So transformation with the OSISAP1 gene raised the salinity-alkali-tolerance of the transgenic plants to a high level.

  3. An embryogenic suspension cell culture system for Agrobacterium-mediated transformation of citrus.

    PubMed

    Dutt, M; Grosser, J W

    2010-11-01

    A method for the genetic transformation of several citrus cultivars is described, including cultivars observed to be recalcitrant to conventional epicotyl-mediated transformation. Embryogenic cell suspension cultures, established from unfertilized ovules were used as target tissues for Agrobacterium-mediated transformation. Several modifications were made to the culture environment to investigate factors required for efficient transfer of the T-DNA and the subsequent regeneration of transgenic citrus plants. It was determined that co-cultivation of citrus cells and Agrobacterium in EME medium supplemented with maltose (EME-M) and 100 μM acetosyringone for 5 days at 25°C was optimum for transformation of each of the citrus cultivars. Efficient selection was obtained and escapes were prevented when the antibiotic hygromycin B was used as a selection antibiotic following transformation with an Agrobacterium strain containing hptII in the T-DNA region. Transgenic embryo regeneration and development was enhanced in medium that contained a liquid overlay consisting of a 1:2 mixture of 0.6 M BH3 and 0.15 M EME-M media. PCR and Southern blot analyses confirmed the presence of the T-DNA and the stable integration into the genome of regenerated plants, while RT-PCR demonstrated variable amounts of RNA being transcribed in different transgenic lines. This protocol can create an avenue for insertion of useful traits into any polyembryonic citrus cultivar that can be established as embryogenic cell suspension cultures, including popular specialty mandarins and seedless cultivars.

  4. High frequency vector-mediated transformation and gene replacement in Tetrahymena.

    PubMed Central

    Gaertig, J; Gu, L; Hai, B; Gorovsky, M A

    1994-01-01

    Recently, we developed a mass DNA-mediated transformation technique for the ciliated protozoan Tetrahymena thermophila that introduces transforming DNA by electroporation into conjugating cells. Other studies demonstrated that a neomycin resistance gene flanked by Tetrahymena H4-I gene regulatory sequences transformed Tetrahymena by homologous recombination within the H4-I locus when microinjected into the macronucleus. We describe the use of conjugant electrotransformation (CET) for gene replacement and for the development of new independently replicating vectors and a gene cassette that can be used as a selectable marker in gene knockout experiments. Using CET, the neomycin resistance gene flanked by H4-I sequences transformed Tetrahymena, resulting in the replacement of the H4-I gene or integrative recombination of the H4-I/neo/H4-I gene (but not vector sequences) in the 5' or 3' flanking region of the H4-I locus. Gene replacement was obtained with non-digested plasmid DNA but releasing the insert increased the frequency of replacement events about 6-fold. The efficiency of transformation by the H4-I/neo/H4-I selectable marker was unchanged when a single copy of the Tetrahymena rDNA replication origin was included on the transforming plasmid. However, the efficiency of transformation using CET increased greatly when a tandem repeat of the replication origin fragment was used. This high frequency of transformation enabled mapping of the region required for H4-I promoter function to within 333 bp upstream of the initiator ATG. Similarly approximately 300 bp of sequence downstream of the translation terminator TGA of the beta-tubulin 2 (BTU2) gene could substitute for the 3' region of the H4-I gene. This hybrid H4-I/neo/BTU2 gene did not transform Tetrahymena when subcloned on a plasmid lacking an origin of replication, but did transform at high frequency on a two origin plasmid. Thus, the H4-I/neo/BTU2 cassette is a selectable marker that can be used for gene

  5. Differences in kinase-mediated regulation of cell cycle progression in normal and transformed cells

    SciTech Connect

    Crissman, H.A.; Gadbois, D.M.; Tobey, R.A.; Stevenson, A.P.; Kraemer, P.M.; Bustos, L.D.; Dickson, J.A.; Bradbury, E.M. )

    1993-01-01

    Staurosporine (Stsp), a general protein kinase inhibitor, was used to investigate the role of kinase-mediated mechanisms in regulating mammalian cell proliferation. Low levels of Stsp (1-2nM) prevented nontransformed cells from entering S phase, indicating that protein phosphorylation processes are essential for commitment of DNA replication in normal cells. Cells resumed cycling when Stsp was removed. The period of sensitivity of nontransformed human diploid fibroblasts to low levels of the drug commenced 3 h later than the G0/G1 boundary and extended through the G1/S boundary. The initial block point at 3 h corresponds neither to the serum nor the amino acid restriction point. In contrast, neither low nor high concentrations (100nm) of Stsp affected G1 progression of transformed cells. High drug concentrations blocked normal cells in G1 and G2 but affected only G2-progression in transformed cells. These results indicate that kinase-mediated regulation of DNA replication is lost as a result of neoplastic transformation, but the G2-arrest mechanism remains intact.

  6. Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava.

    PubMed

    Bull, S E; Owiti, J A; Niklaus, M; Beeching, J R; Gruissem, W; Vanderschuren, H

    2009-01-01

    Agrobacterium-mediated transformation of friable embryogenic calli (FEC) is the most widely used method to generate transgenic cassava plants. However, this approach has proven to be time-consuming and can lead to changes in the morphology and quality of FEC, influencing regeneration capacity and plant health. Here we present a comprehensive, reliable and improved protocol, taking approximately 6 months, that optimizes Agrobacterium-mediated transformation of FEC from cassava model cultivar TMS60444. We cocultivate the FEC with Agrobacterium directly on the propagation medium and adopt the extensive use of plastic mesh for easy and frequent transfer of material to new media. This minimizes stress to the FEC cultures and permits a finely balanced control of nutrients, hormones and antibiotics. A stepwise increase in antibiotic concentration for selection is also used after cocultivation with Agrobacterium to mature the transformed FEC before regeneration. The detailed information given here for each step should enable successful implementation of this technology in other laboratories, including those being established in developing countries where cassava is a staple crop.

  7. Controlling the plasmonic surface waves of metallic nanowires by transformation optics

    SciTech Connect

    Liu, Yichao; Yuan, Jun; Yin, Ge; Ma, Yungui; He, Sailing

    2015-07-06

    In this letter, we introduce the technique of using transformation optics to manipulate the mode states of surface plasmonic waves of metallic nanowire waveguides. As examples we apply this technique to design two optical components: a three-dimensional (3D) electromagnetic mode rotator and a mode convertor. The rotator can rotate the polarization state of the surface wave around plasmonic nanowires by arbitrarily desired angles, and the convertor can transform the surface wave modes from one to another. Full-wave simulation is performed to verify the design and efficiency of our devices. Their potential application in photonic circuits is envisioned.

  8. Steering light by a sub-wavelength metallic grating from transformation optics

    PubMed Central

    Xu, Yadong; Fu, Yangyang; Chen, Huanyang

    2015-01-01

    Transformation optics has shown great ability in designing devices with novel functionalities, such as invisibility cloaking. A recent work shows that it can also be used to design metasurfaces which usually come from the concept of phase discontinuities. However, metasurfaces from transformation optics have very complicated material parameters. Here in this work, we propose a practical design, a sub-wavelength metallic grating with discrete and gradient index materials. Such a design not only inherits some functionalities of metasurfaces from phase discontinuities, but also shows richer physics. Our work will also provide a guidance to recent activities of acoustic metasurfaces, especially for those made of extremely anisotropic metamaterials. PMID:26183391

  9. Steering light by a sub-wavelength metallic grating from transformation optics.

    PubMed

    Xu, Yadong; Fu, Yangyang; Chen, Huanyang

    2015-07-17

    Transformation optics has shown great ability in designing devices with novel functionalities, such as invisibility cloaking. A recent work shows that it can also be used to design metasurfaces which usually come from the concept of phase discontinuities. However, metasurfaces from transformation optics have very complicated material parameters. Here in this work, we propose a practical design, a sub-wavelength metallic grating with discrete and gradient index materials. Such a design not only inherits some functionalities of metasurfaces from phase discontinuities, but also shows richer physics. Our work will also provide a guidance to recent activities of acoustic metasurfaces, especially for those made of extremely anisotropic metamaterials.

  10. Controlled insulator-to-metal transformation in printable polymer composites with nanometal clusters

    NASA Astrophysics Data System (ADS)

    Sivaramakrishnan, Sankaran; Chia, Perq-Jon; Yeo, Yee-Chia; Chua, Lay-Lay; Ho, Peter K.-H.

    2007-02-01

    Although organic semiconductors have received the most attention, the development of compatible passive elements, such as interconnects and electrodes, is also central to plastic electronics. For this, ligand-protected metal-cluster films have been shown to anneal at low temperatures below 250∘C to highly conductive metal films, but they suffer from cracking and inadequate substrate adhesion. Here, we report printable metal-cluster-polymer nanocomposites that anneal to a controlled-percolation nanostructure without complete sintering of the metal clusters. This overcomes the previous challenges while still retaining the desired low transformation temperatures. Highly water- and alcohol-soluble gold clusters (75mgml-1) were synthesized and homogeneously dispersed into poly(3,4-ethylenedioxythiophene) to give a material with annealed d.c. conductivity tuneable between 10-4 and 105Scm-1. These composites can inject holes efficiently into all-printed polymer organic transistors. The insulator-metal transformation can also be electrically induced at 1MVcm-1, suggesting possible memory applications.

  11. Metal complex-based electron-transfer mediators in dye-sensitized solar cells

    DOEpatents

    Elliott, C. Michael; Sapp, Shawn A.; Bignozzi, Carlo Alberto; Contado, Cristiano; Caramori, Stefano

    2006-03-28

    This present invention provides a metal-ligand complex and methods for using and preparing the same. In particular, the metal-ligand complex of the present invention is of the formula: L.sub.a-M-X.sub.b where L, M, X, a, and b are those define herein. The metal-ligand complexes of the present invention are useful in a variety of applications including as electron-transfer mediators in dye-sensitized solar cells and related photoelectrochromic devices.

  12. Selective transformations of cyclopentadienyl ligands of transition-metal and rare-earth metal complexes.

    PubMed

    Liu, Ruiting; Zhou, Xigeng

    2013-04-21

    Cyclopentadienyl and substituted cyclopentadienyl ligands are observed in a wide range of organometallic complexes. In addition to serving as ancillary ligands, these ligands have come into their own as intermediates in organometallic reactions, and shown many unique reaction modes involving ring C-H, C-C and C=C bond cleavages. This feature article summarizes the progressive development of cyclopentadienyl-based reactions of metallocene complexes of transition metals and rare-earth metals, with the aim of further developing the fundamental modes of reactivity of such systems together with their synthetic applications.

  13. Light-Induced In Situ Transformation of Metal Clusters to Metal Nanocrystals for Photocatalysis.

    PubMed

    Xiao, Fang-Xing; Zeng, Zhiping; Hsu, Shao-Hui; Hung, Sung-Fu; Chen, Hao Ming; Liu, Bin

    2015-12-30

    In situ transformation of glutathione-capped gold (Aux) clusters to gold (Au) nanocrystals under simulated solar light irradiation was achieved and utilized as a facile synthetic approach to rationally fabricate Aux/Au/TiO2 ternary and Au/TiO2 binary heterostructures. Synergistic interaction of Aux clusters and Au nanocrystals contributes to enhanced visible-light-driven photocatalysis.

  14. Metal-induced rapid transformation of diamond into single and multilayer graphene on wafer scale

    PubMed Central

    Berman, Diana; Deshmukh, Sanket A.; Narayanan, Badri; Sankaranarayanan, Subramanian K. R. S.; Yan, Zhong; Balandin, Alexander A.; Zinovev, Alexander; Rosenmann, Daniel; Sumant, Anirudha V.

    2016-01-01

    The degradation of intrinsic properties of graphene during the transfer process constitutes a major challenge in graphene device fabrication, stimulating the need for direct growth of graphene on dielectric substrates. Previous attempts of metal-induced transformation of diamond and silicon carbide into graphene suffers from metal contamination and inability to scale graphene growth over large area. Here, we introduce a direct approach to transform polycrystalline diamond into high-quality graphene layers on wafer scale (4 inch in diameter) using a rapid thermal annealing process facilitated by a nickel, Ni thin film catalyst on top. We show that the process can be tuned to grow single or multilayer graphene with good electronic properties. Molecular dynamics simulations elucidate the mechanism of graphene growth on polycrystalline diamond. In addition, we demonstrate the lateral growth of free-standing graphene over micron-sized pre-fabricated holes, opening exciting opportunities for future graphene/diamond-based electronics. PMID:27373740

  15. Metal-induced rapid transformation of diamond into single and multilayer graphene on wafer scale

    SciTech Connect

    Berman, Diana; Deshmukh, Sanket; Narayanan, Badri; Sankaranarayanan, Subramanian K.R.S.; Yan, Zhong; Balandin, Alexander A.; Zinovev, Alexander; Rosenmann, Daniel; Sumant, Anirudha V.

    2016-07-04

    The degradation of intrinsic properties of graphene during the transfer process constitutes a major challenge in graphene device fabrication, stimulating the need for direct growth of graphene on dielectric substrates. Previous attempts of metal-induced transformation of diamond and silicon carbide into graphene suffers from metal contamination and inability to scale graphene growth over large area. Here in this article, we introduce a direct approach to transform polycrystalline diamond into high-quality graphene layers on wafer scale (4 inch in diameter) using a rapid thermal annealing process facilitated by a nickel, Ni thin film catalyst on top. We show that the process can be tuned to grow single or multilayer graphene with good electronic properties. Molecular dynamics simulations elucidate the mechanism of graphene growth on polycrystalline diamond. Additionally, we demonstrate the lateral growth of free-standing graphene over micron-sized pre-fabricated holes, opening exciting opportunities for future graphene/diamond-based electronics.

  16. Metal-induced rapid transformation of diamond into single and multilayer graphene on wafer scale

    NASA Astrophysics Data System (ADS)

    Berman, Diana; Deshmukh, Sanket A.; Narayanan, Badri; Sankaranarayanan, Subramanian K. R. S.; Yan, Zhong; Balandin, Alexander A.; Zinovev, Alexander; Rosenmann, Daniel; Sumant, Anirudha V.

    2016-07-01

    The degradation of intrinsic properties of graphene during the transfer process constitutes a major challenge in graphene device fabrication, stimulating the need for direct growth of graphene on dielectric substrates. Previous attempts of metal-induced transformation of diamond and silicon carbide into graphene suffers from metal contamination and inability to scale graphene growth over large area. Here, we introduce a direct approach to transform polycrystalline diamond into high-quality graphene layers on wafer scale (4 inch in diameter) using a rapid thermal annealing process facilitated by a nickel, Ni thin film catalyst on top. We show that the process can be tuned to grow single or multilayer graphene with good electronic properties. Molecular dynamics simulations elucidate the mechanism of graphene growth on polycrystalline diamond. In addition, we demonstrate the lateral growth of free-standing graphene over micron-sized pre-fabricated holes, opening exciting opportunities for future graphene/diamond-based electronics.

  17. Supramolecular isomers, water clusters and solvent-mediated transformations in a series of lanthanide MOFs

    NASA Astrophysics Data System (ADS)

    Michaelides, Adonis; Skoulika, Stavroula

    2017-09-01

    A series of Lanthanide MOFs of hydromuconic acid (H2hymuc) with cations of different size (La3+, Ce3+, Gd3+, Y3+) were generated at room temperature, pH = 5.5-5.8, using water as solvent. Two microporous genuine supramolecular isomers were found for the larger cations (La3+, Ce3+) sustained either by discrete (H2O)18 clusters or by infinite water tapes, consisting of fused tetramers, pentamers and hexamers. The smaller cations (Gd3+, Y3+) produced a 2D phase, considered as catenane isomer of previously published lanthanide MOFs with adipic acid. MOFs obtained with Ce3+, Gd3+and Y3+ undergo irreversible solvent-mediated transformation, yielding isomorphous 3D two-fold interpenetrated MOFs. The close structural similarity between the 2D phase and the 3D interpenetrated one indicated a possible mechanism for this transformation.

  18. A simple and highly efficient Agrobacterium-mediated transformation protocol for Setaria viridis.

    PubMed

    Martins, Polyana Kelly; Ribeiro, Ana Paula; Cunha, Bárbara Andrade Dias Brito da; Kobayashi, Adilson Kenji; Molinari, Hugo Bruno Correa

    2015-06-01

    The production and use of sugarcane in Brazil is very important for bioenergy production and is recognized as one of the most efficient in the world. In our laboratory, Setaria viridis is being tested as a model plant for sugarcane. S. viridis has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model system. We report a highly efficient protocol for Agrobacterium-mediated genetic transformation of S. viridis. The optimization of several steps in tissue culture allowed the rapid regeneration of plants and increased the rate of transformation up to 29%. This protocol could become a powerful tool for functional genomics in sugarcane.

  19. Agrobacterium-mediated transformation of Malus robusta with tomato iron transporter gene.

    PubMed

    Qu, Shen-Chun; Huang, Xiao-De; Zhang, Zhen; Yao, Quan-Hong; Tao, Jian-Min; Qiao, Yu-Shan; Zhang, Jun-Yi

    2005-06-01

    The tomato iron transporter gene (LeIRT2) was introduced to Malus robusta Rehd. via Agrobacterium-mediated transformation to produce iron-deficiency tolerant apple rootstock. A total of 19 putative transformants were obtained, 11 of which were verified by PCR amplification to carry a fragment of the transgene. Among them, nine were confirmed to carry the transgene by Southern blot analysis with one to three copies of the transgene integrated into the plant genome. Two transgenic plants, one carrying one copy and the other three copies of the transgene, were hydroponically cultured to test their tolerance to iron-deficiency, which was found only in the transgenic plant with a single copy, which weighted 21%-4% greater than those of the control plants.

  20. Electric field mediated transformation: isolation and characterization of a TK+ subclone.

    PubMed

    Zerbib, D; Amalric, F; Teissié, J

    1985-06-28

    Transformation of mammalian TK- cells by a plasmid carrying the TK gene from Herpes virus simplex 1 (pAGO) was mediated by electroporation. The cells were treated either in suspension or growing in monolayers directly in the petri dish. The yield of transformation was between 8.10(-5) and 2.10(-4) per microgram DNA depending on the experimental conditions. The structure of the integrated DNA was investigated proving the occurrence of a duplication process that affected preferentially the pBR322 part of the pAGO DNA (60 copies per cell). The TK gene that gave the TK+ phenotype to the selected clone was present in less than 6 copies.

  1. Deletion of host histone acetyltransferases and deacetylases strongly affects Agrobacterium-mediated transformation of Saccharomyces cerevisiae.

    PubMed

    Soltani, Jalal; van Heusden, Gerard Paul H; Hooykaas, Paul J J

    2009-09-01

    Agrobacterium tumefaciens is a plant pathogen that genetically transforms plant cells by transferring a part of its Ti-plasmid, the T-strand, to the host cell. Under laboratory conditions, it can also transform cells from many different nonplant organisms, including the yeast Saccharomyces cerevisiae. Collections of S. cerevisiae strains have been developed with systematic deletion of all coding sequences. Here, we used these collections to identify genes involved in the Agrobacterium-mediated transformation (AMT) of S. cerevisiae. We found that deletion of genes (GCN5, NGG1, YAF9 and EAF7) encoding subunits of the SAGA, SLIK, ADA and NuA4 histone acetyltransferase complexes highly increased the efficiency of AMT, while deletion of genes (HDA2, HDA3 and HST4) encoding subunits of histone deacetylase complexes decreased AMT. These effects are specific for AMT as the efficiency of chemical (lithium acetate) transformation was not or only slightly affected by these deletions. Our data are consistent with a positive role of host histone deacetylation in AMT.

  2. Agrobacterium-mediated genetic transformation of pineapple (Ananas comosus L., Merr.).

    PubMed

    Mhatre, Minal

    2013-01-01

    Pineapple (Ananas comosus L., Merr.) is a commercially important crop, grown in the tropical and subtropical regions. However, the crop is faced with postharvest damage and poor varietal and nutritional improvement. Being a vegetatively propagated crop, conventional breeding programs take longer time for genetic improvement, which may not necessarily successfully develop an improved cultivar. Hence, the genetic modification of pineapple is an alternative handy approach to improve pineapple. We have established an Agrobacterium-mediated transformation system using leaf bases from in vitro-grown pineapple plants. Being a monocot, acetosyringone is added to the culture medium for overnight growth of Agrobacterium and transformation to transfer a gene of interest MSI99 soybean ferritin. Leaf bases isolated from in vitro shoot cultures are treated with Agrobacterium suspension at two dilutions, 10× and 20×, for 30 min. Explants are subsequently blot dried and cultured on gelrite solidified hormone-free Pin1 medium for 2 days (cocultivation). Periodic transfer is first done to the regeneration medium (Pin1) containing cefotaxime for the suppression of Agrobacterium growth. The transformants are selected by culturing on Pin1 medium containing cefotaxime and kanamycin. Multiple shoots, regenerated in leaf bases, are further multiplied and individually rooted in the liquid RM medium amended with antibiotics to recover plants. Putative transformants are analyzed for transgene integration and expression using standard molecular biological methods of PCR, RT-PCR, and genomic Southern.

  3. Agrobacterium-mediated transformation of embryogenic cultures and plant regeneration in Vitis rotundifolia Michx. (muscadine grape).

    PubMed

    Dhekney, S A; Li, Z T; Dutt, M; Gray, D J

    2008-05-01

    A method to produce transgenic plants of Vitis rotundifolia was developed. Embryogenic cultures were initiated from leaves of in vitro grown shoot cultures and used as target tissues for Agrobacterium-mediated genetic transformation. A green fluorescent protein/neomycin phosphotransferase II (gfp/nptII) fusion gene that allowed for simultaneous selection of transgenic cells based on GFP fluorescence and kanamycin resistance was used to optimize parameters influencing genetic transformation. It was determined that both proembryonal masses (PEM) and mid-cotyledonary stage somatic embryos (SE) were suitable target tissues for co-cultivation with Agrobacterium as evidenced by transient GFP expression. Kanamycin at 100 mg l(-1) in the culture medium was effective in suppression of non-transformed tissue and permitting the growth and development of transgenic cells, compared to 50 or 75 mg l(-1), which permitted the proliferation of more non-transformed cells. Transgenic plants of "Alachua" and "Carlos" were recovered after secondary somatic embryogenesis from primary SE explants co-cultivated with Agrobacterium. The presence and stable integration of transgenes in transgenic plants was confirmed by PCR and Southern blot hybridization. Transgenic plants exhibited uniform GFP expression in cells of all plant tissues and organs including leaves, stems, roots, inflorescences and the embryo and endosperm of developing berries.

  4. A link between c-Myc-mediated transcriptional repression and neoplastic transformation.

    PubMed Central

    Lee, L A; Dolde, C; Barrett, J; Wu, C S; Dang, C V

    1996-01-01

    Recent studies indicate that the transcription factor c-Myc contributes to oncogenesis by altering the expression of genes involved in cell proliferation, but its precise function in neoplasia remains ambiguous. The ability of c-Myc to bind the sequence CAC(G/A)TG and transactivate appears to be linked to its transforming activity; however, c-Myc also represses transcription in vitro through a pyrimidine-rich cis element termed the initiator (Inr). In transfection experiments using the adenoviral major late (adML) promoter, which contains two Myc binding sites and an Inr, we determined that c-Myc represses transcription through the initiator in vivo. This activity requires the dimerization domain and amino acids 106 to 143, which are located within the transactivation domain and are necessary for neoplastic transformation. We studied a lymphoma-derived c-Myc substitution mutation at 115-Phe, which is within the region required for transcriptional suppression, and found the mutant more effective than wild-type c-Myc in transforming rodent fibroblasts and in suppressing the adML promoter. Our studies of both loss-of-function and gain-of-function c-Myc mutations suggest a link between c-Myc-mediated neoplastic transformation and transcriptional repression through the Inr. PMID:8601634

  5. A link between c-Myc-mediated transcriptional repression and neoplastic transformation.

    PubMed

    Lee, L A; Dolde, C; Barrett, J; Wu, C S; Dang, C V

    1996-04-01

    Recent studies indicate that the transcription factor c-Myc contributes to oncogenesis by altering the expression of genes involved in cell proliferation, but its precise function in neoplasia remains ambiguous. The ability of c-Myc to bind the sequence CAC(G/A)TG and transactivate appears to be linked to its transforming activity; however, c-Myc also represses transcription in vitro through a pyrimidine-rich cis element termed the initiator (Inr). In transfection experiments using the adenoviral major late (adML) promoter, which contains two Myc binding sites and an Inr, we determined that c-Myc represses transcription through the initiator in vivo. This activity requires the dimerization domain and amino acids 106 to 143, which are located within the transactivation domain and are necessary for neoplastic transformation. We studied a lymphoma-derived c-Myc substitution mutation at 115-Phe, which is within the region required for transcriptional suppression, and found the mutant more effective than wild-type c-Myc in transforming rodent fibroblasts and in suppressing the adML promoter. Our studies of both loss-of-function and gain-of-function c-Myc mutations suggest a link between c-Myc-mediated neoplastic transformation and transcriptional repression through the Inr.

  6. AGROBACTERIUM-MEDIATED TRANSFORMATION IN THE GREEN ALGA HAEMATOCOCCUS PLUVIALIS (CHLOROPHYCEAE, VOLVOCALES)(1).

    PubMed

    Kathiresan, S; Chandrashekar, A; Ravishankar, G A; Sarada, R

    2009-06-01

    The first successful Agrobacterium-mediated transformation of the green alga Haematococcus pluvialis Flot. using the binary vectors hosting the genes coding for GUS (β-glucuronidase), GFP (green fluorescent protein), and hpt (hygromycin phosphotransferase) is reported here. Colonies resistant to hygromycin at 10 mg · L(-1) expressed β-glucuronidase. The greenish yellow fluorescence of GFP was observed when the hygromycin-resistant cells were viewed with a fluorescent microscope. PCR was used to successfully amplify fragments of the hpt (407 bp) and GUS (515 bp) genes from transformed cells, while Southern blots indicated the integration of the hygromycin gene into the genome of H. pluvialis. SEM indicated that the cell wall of H. pluvialis was altered on infection with Agrobacterium. The transformation achieved here by Agrobacterium does not need treatment with acetosyringone or the wounding of cells. A robust transformation method for this alga would pave the way for manipulation of many important pathways relevant to the food, pharmaceutical, and nutraceutical industries.

  7. Efficient soybean regeneration and Agrobacterium-mediated transformation using a whole cotyledonary node as an explant.

    PubMed

    Zhang, Fuli; Chen, Can; Ge, Honglian; Liu, Jinmei; Luo, Yunling; Liu, Kun; Chen, Long; Xu, Kedong; Zhang, Yi; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    An optimized regeneration and Agrobacterium-mediated transformation protocol based on whole cotyledonary node explants was developed in soybean (Glycine max) cultivar Zhong Huang 13. Adding 6-benzylaminopurine (BAP) in a germinating medium could significantly increase regeneration efficiency; the optimal BAP concentration for shoot formation was 0.5 mg/L. The concentrations of plant growth regulators in a shoot induction medium were optimized by the orthogonal test [L9 (3(3))]. The best combination for shoot regeneration was a medium of Murashige & Skoog salts with B5 vitamins (MSB) supplemented with 3.5 mg/L BAP, 0.2 mg/L indole-3-butyric acid (IBA), and 0.2 mg/L kinetin (KT). Under this favorable condition, one node could regenerate 28-30 shoots. Soybean whole cotyledonary nodes were transformed by inoculation with A. tumefaciens strain EHA105 harboring a vector pBI121 containing a β-glucuronidase gene (gus). GUS assay, polymerase chain reaction, and Southern blot analysis indicated that the gus gene was transformed into soybean plants with 23.1% transformation efficiency. Transgenic plants could be obtained within 5-6 weeks, which was about 4 weeks less than that of a traditional single cotyledonary node method. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  8. [Agrobacterium-mediated transformation of LJAMP2 gene into 'Red Sun' kiwifruit and its molecular identification].

    PubMed

    Zhou, Yue; Zhao, Xupeng; Wu, Xiuhua; Zhang, Yanling; Zhang, Lin; Luo, Keming; Tang, Shaohu

    2014-06-01

    Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.

  9. Controlled Encapsulation of Functional Organic Molecules within Metal-Organic Frameworks: In Situ Crystalline Structure Transformation.

    PubMed

    Guan, Jinju; Hu, Yu; Wang, Yu; Li, Hongfeng; Xu, Zhiling; Zhang, Tao; Wu, Peng; Zhang, Suoying; Xiao, Gengwu; Ji, Wenlan; Li, Linjie; Zhang, Meixuan; Fan, Yun; Li, Lin; Zheng, Bing; Zhang, Weina; Huang, Wei; Huo, Fengwei

    2017-01-23

    Functional organic molecules/metal-organic frameworks composites can be obtained by in situ crystalline structure transformation from ZIF-L to ZIF-8-L under double solvent conditions. Interestingly, the as-prepared molecules/ZIF-8-L composites with the leaf-like morphology exhibit good fluorescence properties and size selectivity in fluorescent quenchers due to the molecular sieving effect of the well-defined microporous ZIF-8-L.

  10. Recovery of Olfactory Mediated Behaviours of Fish from Metal Contaminated Lakes.

    PubMed

    Azizishirazi, Ali; Pyle, Greg G

    2015-07-01

    Fish mediate many biological processes by olfaction, which can be impaired by contaminants (i.e. metals). While the olfactory recovery of fish from metal contaminated lakes if subsequently cultured in clean water has been shown at the neurophysiological level, the recovery potential of olfactory mediated behaviours remains unknown. To study behavioural recovery of fish from metal contaminated lakes, wild yellow perch (Perca flavescens) were collected from two metal-contaminated lakes (Ramsey and Hannah lakes) in the metal-mining district of Sudbury, ON, Canada and cultured in clean water from a reference lake (Geneva Lake) for another 24 h. Olfactory mediated behaviours of the test organisms were tested using avoidance responses to conspecific skin extract. While olfactory mediated behaviours of fish from Ramsey Lake (low contamination) recovered after 24 h in clean water, recovery could not be observed in fish from Hannah Lake (high contamination). These results demonstrate that the recovery of behavioural deficits of fish from metal contaminated lakes is depending on the habitats' metal concentration.

  11. Transforming growth factor-beta1 mediates cellular response to DNA damage in situ

    NASA Technical Reports Server (NTRS)

    Ewan, Kenneth B.; Henshall-Powell, Rhonda L.; Ravani, Shraddha A.; Pajares, Maria Jose; Arteaga, Carlos; Warters, Ray; Akhurst, Rosemary J.; Barcellos-Hoff, Mary Helen

    2002-01-01

    Transforming growth factor (TGF)-beta1 is rapidly activated after ionizing radiation, but its specific role in cellular responses to DNA damage is not known. Here we use Tgfbeta1 knockout mice to show that radiation-induced apoptotic response is TGF-beta1 dependent in the mammary epithelium, and that both apoptosis and inhibition of proliferation in response to DNA damage decrease as a function of TGF-beta1 gene dose in embryonic epithelial tissues. Because apoptosis in these tissues has been shown previously to be p53 dependent, we then examined p53 protein activation. TGF-beta1 depletion, by either gene knockout or by using TGF-beta neutralizing antibodies, resulted in decreased p53 Ser-18 phosphorylation in irradiated mammary gland. These data indicate that TGF-beta1 is essential for rapid p53-mediated cellular responses that mediate cell fate decisions in situ.

  12. Transforming growth factor-beta1 mediates cellular response to DNA damage in situ

    NASA Technical Reports Server (NTRS)

    Ewan, Kenneth B.; Henshall-Powell, Rhonda L.; Ravani, Shraddha A.; Pajares, Maria Jose; Arteaga, Carlos; Warters, Ray; Akhurst, Rosemary J.; Barcellos-Hoff, Mary Helen

    2002-01-01

    Transforming growth factor (TGF)-beta1 is rapidly activated after ionizing radiation, but its specific role in cellular responses to DNA damage is not known. Here we use Tgfbeta1 knockout mice to show that radiation-induced apoptotic response is TGF-beta1 dependent in the mammary epithelium, and that both apoptosis and inhibition of proliferation in response to DNA damage decrease as a function of TGF-beta1 gene dose in embryonic epithelial tissues. Because apoptosis in these tissues has been shown previously to be p53 dependent, we then examined p53 protein activation. TGF-beta1 depletion, by either gene knockout or by using TGF-beta neutralizing antibodies, resulted in decreased p53 Ser-18 phosphorylation in irradiated mammary gland. These data indicate that TGF-beta1 is essential for rapid p53-mediated cellular responses that mediate cell fate decisions in situ.

  13. [Effects of stabilization treatment on migration and transformation of heavy metals in mineral waste residues].

    PubMed

    Zhao, Shu-Hua; Chen, Zhi-Liang; Zhang, Tai-Ping; Pan, Wei-Bin; Peng, Xiao-Chun; Che, Rong; Ou, Ying-Juan; Lei, Guo-Jian; Zhou, Ding

    2014-04-01

    Different forms of heavy metals in soil will produce different environmental effects, and will directly influence the toxicity, migration and bioavailability of heavy metals. This study used lime, fly ash, dried sludge, peanut shells as stabilizers in the treatment of heavy metals in mineral waste residues. Morphological analyses of heavy metal, leaching experiments, potted plant experiments were carried out to analyze the migration and transformation of heavy metals. The results showed that after adding stabilizers, the pH of the acidic mineral waste residues increased to more than neutral, and the organic matter content increased significantly. The main existing forms of As, Pb, and Zn in the mineral waste residues were the residual. The contents of exchangeable and organic matter-bound As decreased by 65.6% and 87.7% respectively after adding fly ash, dried sludge and peanut shells. Adding lime, fly ash and peanut shells promoted the transformation of As from the Fe-Mn oxide-bound to the carbonate-bound, and adding lime and fly ash promoted the transformation of Pb and Zn from the exchangeable, Fe-Mn oxide-bound, organic matter-bound to the residual. After the early stage of the stabilization treatment, the contents of As, Pb and Zn in the leachate had varying degrees of decline, and adding peanut shells could reduce the contents of As, Pb and Zn in the leachate further. Among them, the content of As decreased most significantly after treatment with fly ash, dried sludge and peanut shells, with a decline of 57.4%. After treatment with lime, fly ash and peanut shells, the content of Zn decreased most significantly, by 24.9%. The addition of stabilizers was advantageous to the germination and growth of plants. The combination of fly ash, dried sludge and peanut shell produced the best effect, and the Vetiveria zizanioides germination rate reached 76% in the treated wasted mineral residues.

  14. A rapid, highly efficient and economical method of Agrobacterium-mediated in planta transient transformation in living onion epidermis.

    PubMed

    Xu, Kedong; Huang, Xiaohui; Wu, Manman; Wang, Yan; Chang, Yunxia; Liu, Kun; Zhang, Ju; Zhang, Yi; Zhang, Fuli; Yi, Liming; Li, Tingting; Wang, Ruiyue; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The often used transient transformation methods included particle bombardment, protoplast transfection and Agrobacterium-mediated transformation. Particle bombardment in onion epidermis was successfully established, however, it was expensive, biolistic equipment dependent and with low transformation efficiency. We developed a highly efficient in planta transient transformation method in onion epidermis by using a special agroinfiltration method, which could be fulfilled within 5 days from the pretreatment of onion bulb to the best time-point for analyzing gene expression. The transformation conditions were optimized to achieve 43.87% transformation efficiency in living onion epidermis. The developed method has advantages in cost, time-consuming, equipment dependency and transformation efficiency in contrast with those methods of particle bombardment in onion epidermal cells, protoplast transfection and Agrobacterium-mediated transient transformation in leaf epidermal cells of other plants. It will facilitate the analysis of protein subcellular localization on a large scale.

  15. A Rapid, Highly Efficient and Economical Method of Agrobacterium-Mediated In planta Transient Transformation in Living Onion Epidermis

    PubMed Central

    Xu, Kedong; Huang, Xiaohui; Wu, Manman; Wang, Yan; Chang, Yunxia; Liu, Kun; Zhang, Ju; Zhang, Yi; Zhang, Fuli; Yi, Liming; Li, Tingting; Wang, Ruiyue; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The often used transient transformation methods included particle bombardment, protoplast transfection and Agrobacterium-mediated transformation. Particle bombardment in onion epidermis was successfully established, however, it was expensive, biolistic equipment dependent and with low transformation efficiency. We developed a highly efficient in planta transient transformation method in onion epidermis by using a special agroinfiltration method, which could be fulfilled within 5 days from the pretreatment of onion bulb to the best time-point for analyzing gene expression. The transformation conditions were optimized to achieve 43.87% transformation efficiency in living onion epidermis. The developed method has advantages in cost, time-consuming, equipment dependency and transformation efficiency in contrast with those methods of particle bombardment in onion epidermal cells, protoplast transfection and Agrobacterium-mediated transient transformation in leaf epidermal cells of other plants. It will facilitate the analysis of protein subcellular localization on a large scale. PMID:24416168

  16. Improvement of Agrobacterium-mediated transformation in Hi-II maize (Zea mays) using standard binary vectors.

    PubMed

    Vega, Juan M; Yu, Weichang; Kennon, Angela R; Chen, Xinlu; Zhang, Zhanyuan J

    2008-02-01

    High-frequency transformation of maize (Zea mays L.) using standard binary vectors is advantageous for functional genomics and other genetic engineering studies. Recent advances in Agrobacterium tumefaciens-mediated transformation of maize have made it possible for the public to transform maize using standard binary vectors without a need of the superbinary vector. While maize Hi-II has been a preferred maize genotype to use in various maize transformation efforts, there is still potential and need in further improving its transformation frequency. Here we report the enhanced Agrobacterium-mediated transformation of immature zygotic embryos of maize Hi-II using standard binary vectors. This improved transformation process employs low-salt media in combined use with antioxidant L-cysteine alone or L-cysteine and dithiothreitol (DTT) during the Agrobacterium infection stage. Three levels of N6 medium salts, 10, 50, and 100%, were tested. Both 10 and 50% salts were found to enhance the T-DNA transfer in Hi-II. Addition of DTT to the cocultivation medium also improves the T-DNA transformation. About 12% overall and the highest average of 18% transformation frequencies were achieved from a large number of experiments using immature embryos grown in various seasons. The enhanced transformation protocol established here will be advantageous for maize genetic engineering studies including transformation-based functional genomics.

  17. Pressure-induced Transformations of Dense Carbonyl Sulfide to Singly Bonded Amorphous Metallic Solid

    NASA Astrophysics Data System (ADS)

    Kim, Minseob; Dias, Ranga; Ohishi, Yasuo; Matsuoka, Takehiro; Chen, Jing-Yin; Yoo, Choong-Shik

    2016-08-01

    The application of pressure, internal or external, transforms molecular solids into non-molecular extended network solids with diverse crystal structures and electronic properties. These transformations can be understood in terms of pressure-induced electron delocalization; however, the governing mechanisms are complex because of strong lattice strains, phase metastability and path dependent phase behaviors. Here, we present the pressure-induced transformations of linear OCS (R3m, Phase I) to bent OCS (Cm, Phase II) at 9 GPa an amorphous, one-dimensional (1D) polymer at 20 GPa (Phase III); and an extended 3D network above ~35 GPa (Phase IV) that metallizes at ~105 GPa. These results underscore the significance of long-range dipole interactions in dense OCS, leading to an extended molecular alloy that can be considered a chemical intermediate of its two end members, CO2 and CS2.

  18. Pressure-induced Transformations of Dense Carbonyl Sulfide to Singly Bonded Amorphous Metallic Solid

    PubMed Central

    Kim, Minseob; Dias, Ranga; Ohishi, Yasuo; Matsuoka, Takehiro; Chen, Jing-Yin; Yoo, Choong-Shik

    2016-01-01

    The application of pressure, internal or external, transforms molecular solids into non-molecular extended network solids with diverse crystal structures and electronic properties. These transformations can be understood in terms of pressure-induced electron delocalization; however, the governing mechanisms are complex because of strong lattice strains, phase metastability and path dependent phase behaviors. Here, we present the pressure-induced transformations of linear OCS (R3m, Phase I) to bent OCS (Cm, Phase II) at 9 GPa; an amorphous, one-dimensional (1D) polymer at 20 GPa (Phase III); and an extended 3D network above ~35 GPa (Phase IV) that metallizes at ~105 GPa. These results underscore the significance of long-range dipole interactions in dense OCS, leading to an extended molecular alloy that can be considered a chemical intermediate of its two end members, CO2 and CS2. PMID:27527241

  19. Carbonyl mediated conductance through metal bound peptides: a computational study

    NASA Astrophysics Data System (ADS)

    Perrine, Trilisa M.; Dunietz, Barry D.

    2007-10-01

    Large increases in the conductance of peptides upon binding to metal ions have recently been reported experimentally. The mechanism of the conductance switching is examined computationally. It is suggested that oxidation of the metal ion occurs after binding to the peptide. This is caused by the bias potential placed across the metal-peptide complex. A combination of configurational changes, metal ion involvement and interactions between carbonyl group oxygen atoms and the gold leads are all shown to be necessary for the large improvement in the conductance seen experimentally. Differences in the molecular orbitals of the nickel and copper complexes are noted and serve to explain the variation of the improvement in conductance upon binding to either a nickel or copper ion.

  20. Seeded growth of metal-doped plasmonic oxide heterodimer nanocrystals and their chemical transformation.

    PubMed

    Ye, Xingchen; Reifsnyder Hickey, Danielle; Fei, Jiayang; Diroll, Benjamin T; Paik, Taejong; Chen, Jun; Murray, Christopher B

    2014-04-02

    We have developed a generalized seeded-growth methodology for the synthesis of monodisperse metal-doped plasmonic oxide heterodimer nanocrystals (NCs) with a near-unity morphological yield. Using indium-doped cadmium oxide (ICO) as an example, we show that a wide variety of preformed metal NCs (Au, Pt, Pd, FePt, etc.) can serve as the seeds for the tailored synthesis of metal-ICO heterodimers with exquisite size, shape, and composition control, facilitated by the delayed nucleation mechanism of the CdO phase. The metal-ICO heterodimers exhibit broadly tunable near-infrared localized surface plasmon resonances, and dual plasmonic bands are observed for Au-ICO heterodimers. We further demonstrate that the oxide domain of the Au-ICO heterodimers can be selectively and controllably transformed into a series of partially and completely hollow cadmium chalcogenide nanoarchitectures with unprecedented structural complexity, leaving the metal domain intact. Our work not only represents an exciting addition to the rapidly expanding library of chemical reactions that produce colloidal hybrid NCs, but it also provides a general route for the bottom-up chemical design of multicomponent metal-oxide-semiconductor NCs in a rational and sequential manner.

  1. The role of microRNAs in metal carcinogen-induced cell malignant transformation and tumorigenesis.

    PubMed

    Humphries, Brock; Wang, Zhishan; Yang, Chengfeng

    2016-12-01

    MicroRNAs (miRNAs), an important component of epigenetic mechanisms of carcinogenesis, have been shown to play crucial roles in cancer initiation, metastasis, prognosis and responses to drug treatment and may serve as biomarkers for early diagnosis of cancer and tools for cancer therapy. Metal carcinogens, such as arsenic, cadmium, hexavalent chromium and nickel, are well-established human carcinogens causing various cancers upon long term exposure. However, the mechanism of metal carcinogenesis has not been well understood, which limits our capability to effectively diagnose and treat human cancers resulting from chronic metal carcinogen exposure. Over recent years, the role of miRNAs in metal carcinogenesis has been actively explored and a growing body of evidence indicates the critical involvement of miRNAs in metal carcinogenesis. This review aims to discuss recent studies showing that miRNAs play important roles in metal carcinogen-induced cell malignant transformation and tumorigenesis. Some thoughts for future further studies in this field are also presented. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Role of Oxalic Acid Overexcretion in Transformations of Toxic Metal Minerals by Beauveria caledonica

    PubMed Central

    Fomina, M.; Hillier, S.; Charnock, J. M.; Melville, K.; Alexander, I. J.; Gadd, G. M.

    2005-01-01

    The fungus Beauveria caledonica was highly tolerant to toxic metals and solubilized cadmium, copper, lead, and zinc minerals, converting them into oxalates. This fungus was found to overexcrete organic acids with strong metal-chelating properties (oxalic and citric acids), suggesting that a ligand-promoted mechanism was the main mechanism of mineral dissolution. Our data also suggested that oxalic acid was the main mineral-transforming agent. Cadmium, copper, and zinc oxalates were precipitated by the fungus in the local environment and also in association with the mycelium. The presence of toxic metal minerals often led to the formation of mycelial cords, and in the presence of copper-containing minerals, these cords exhibited enhanced excretion of oxalic acid, which resulted in considerable encrustation of the cords by copper oxalate hydrate (moolooite). It was found that B. caledonica hyphae and cords were covered by a thick hydrated mucilaginous sheath which provided a microenvironment for chemical reactions, crystal deposition, and growth. Cryo-scanning electron microscopy revealed that mycogenic metal oxalates overgrew parental fungal hyphae, leaving a labyrinth of fungal tunnels within the newly formed mineral matter. X-ray absorption spectroscopy revealed that oxygen ligands played a major role in metal coordination within the fungal biomass during the accumulation of mobilized toxic metals by B. caledonica mycelium; these ligands were carboxylic groups in copper phosphate-containing medium and phosphate groups in pyromorphite-containing medium. PMID:15640211

  3. Environmental assessment of heavy metal transport and transformation in the Hangzhou Bay, China.

    PubMed

    Fang, Hongwei; Huang, Lei; Wang, Jingyu; He, Guojian; Reible, Danny

    2016-01-25

    The environmental impact of heavy metal (Cu, Cd, Zn, Pb, Ni, (90)Sr and (137)Cs) transport and transformation in the Hangzhou Bay (China) was assessed through a comprehensive model that integrates hydrodynamics, sediment and heavy metal transport. A mechanistic surface complexation model was used to estimate the adsorption and desorption of heavy metal by suspended sediment under different aqueous chemistry conditions. The dynamics of metal exchange to and from the seabed was also assessed. The primary processes regulating heavy metal distribution, i.e., convection-diffusion, adsorption-desorption, sedimentation-resuspension, as well as other physical and chemical processes related to mass exchange between adjacent sediment layers, were considered in detail. The accidental discharge of (137)Cs was simulated as an example and results showed that (137)Cs transported along the coast driven by tidal flow. Most (137)Cs distributed near the outfall and accumulated in the seabed sediment. The proposed model can be a useful tool for predicting heavy metal transport and fate and provide a theoretical basis to guide field sampling, assessment of risks and the design of remediation strategies.

  4. Geomycology: biogeochemical transformations of rocks, minerals, metals and radionuclides by fungi, bioweathering and bioremediation.

    PubMed

    Gadd, Geoffrey M

    2007-01-01

    The study of the role that fungi have played and are playing in fundamental geological processes can be termed 'geomycology' and this article seeks to emphasize the fundamental importance of fungi in several key areas. These include organic and inorganic transformations and element cycling, rock and mineral transformations, bioweathering, mycogenic mineral formation, fungal-clay interactions, metal-fungal interactions, and the significance of such processes in the environment and their relevance to areas of environmental biotechnology such as bioremediation. Fungi are intimately involved in biogeochemical transformations at local and global scales, and although such transformations occur in both aquatic and terrestrial habitats, it is the latter environment where fungi probably have the greatest influence. Within terrestrial aerobic ecosystems, fungi may exert an especially profound influence on biogeochemical processes, particularly when considering soil, rock and mineral surfaces, and the plant root-soil interface. The geochemical transformations that take place can influence plant productivity and the mobility of toxic elements and substances, and are therefore of considerable socio-economic relevance, including human health. Of special significance are the mutualistic symbioses, lichens and mycorrhizas. Some of the fungal transformations discussed have beneficial applications in environmental biotechnology, e.g. in metal leaching, recovery and detoxification, and xenobiotic and organic pollutant degradation. They may also result in adverse effects when these processes are associated with the degradation of foodstuffs, natural products, and building materials, including wood, stone and concrete. It is clear that a multidisciplinary approach is essential to understand fully all the phenomena encompassed within geomycology, and it is hoped that this review will serve to catalyse further research, as well as stimulate interest in an area of mycology of global

  5. Visible Light Induced Organic Transformations Using Metal-Organic-Frameworks (MOFs).

    PubMed

    Deng, Xiaoyu; Li, Zhaohui; García, Hermenegildo

    2017-08-22

    With the aim of developing renewable energy based processes, researchers are paying increasing interest to light induced organic transformations. Metal-organic frameworks (MOFs), a class of micro-/mesoporous hybrid materials, are recently emerging as a new type of photoactive materials for organic syntheses due to their unique structural characteristics. In this Review, we summarized the recent applications of MOFs as photocatalysts for light induced organic transformations, including (1) oxidation of alcohols, amines, alkene, alkanes and sulfides; (2) hydroxylation of aromatic compounds like benzene; (3) activation of the C-H bonds to construct new C-C or C-X bonds; (4) atom-transfer radical polymerization (ATRP). This Review starts with general background information of using MOFs in photocatalysis, followed by a description of light induced organic transformations promoted by photoactive inorganic nodes and photocatalytic active ligands in MOFs, respectively. Thereafter, the use of MOFs as multifunctional catalysts for light induced organic transformations via an efficient merge of the metal/ligand/guest based catalysis where the photocatalytic activity of MOFs plays a key role are discussed. Finally, the limitations, challenges and the future perspective of the application of MOFs for light induced organic transformations were addressed. The objective of this Review is to serve as a starting point for other researchers to get into this largely unexplored field. It is also our goal to stimulate intensive research in this field for rational designing of MOF materials to overcome their current limitations in photocatalysis, which can lead to more creative visible-light-induced organic transformations. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Mineral transformations during the dissolution of uranium ore minerals by dissimilatory metal-reducing bacteria

    NASA Astrophysics Data System (ADS)

    Glasauer, S.; Weidler, P.; Fakra, S.; Tyliszczak, T.; Shuh, D.

    2011-12-01

    Carnotite minerals [X2(UO2)2(VO4)2]; X = K, Ca, Ba, Mn, Na, Cu or Pb] form the major ore of uranium in the Colorado Plateau. These deposits are highly oxidized and contain U(VI) and V(IV). The biotransformation of U(VI) bound in carnotite by bacteria during dissimilatory metal reduction presents a complex puzzle in mineral chemistry. Both U(VI) and V(V) can be respired by metal reducing bacteria, and the mineral structure can change depending on the associated counterion. We incubated anaerobic cultures of S. putrefaciens CN32 with natural carnotite minerals from southeastern Utah in a nutrient-limited defined medium. Strain CN32 is a gram negative bacterium and a terrestrial isolate from New Mexico. The mineral and metal transformations were compared to a system that contained similar concentrations of soluble U(VI) and V(V). Electron (SEM, TEM) microscopies and x-ray spectromicroscopy (STXM) were used in conjunction with XRD to track mineral changes, and bacterial survival was monitored throughout the incubations. Slow rates of metal reduction over 10 months for the treatment with carnotite minerals revealed distinct biotic and abiotic processes, providing insight on mineral transformation and bacteria-metal interactions. The bacteria existed as small flocs or individual cells attached to the mineral phase, but did not adsorb soluble U or V, and accumulated very little of the biominerals. Reduction of mineral V(V) necessarily led to a dismantling of the carnotite structure. Bioreduction of V(V) by CN32 contributed small but profound changes to the mineral system, resulting in new minerals. Abiotic cation exchange within the carnotite group minerals induced the rearrangement of the mineral structures, leading to further mineral transformation. In contrast, bacteria survival was poor for treatments with soluble U(VI) and V(V), although both metals were reduced completely and formed solid UO2 and VO2; we also detected V(III). For these treatments, the bacteria

  7. Towards metal-mediated g-quartet analogues: 1,2,4-triazole nucleotides.

    PubMed

    Withers, Jamie M; Telfer, Shane G; Filichev, Vyacheslav V

    2015-01-01

    We proposed that metal-coordinating nucleotides could be used to control the assembly of G-quadruplexes through the formation of an artificial metal-centered quartet. Several guanine-rich DNA sequences containing 1,2,4-triazole-functionalized nucleotides were investigated. These oligonucleotides were designed to form quartets mediated by metal-triazole bonding both on the surface of and within the G-quadruplex core. In contrast to duplex studies in which 1,2,4-triazole nucleosides serve as a mimic of Watson-Crick base-pairs, our results show that these nucleosides are not suitable components of an artificial metal-centered quartet.

  8. Metal nanodot arrays fabricated via seed-mediated electroless plating with block copolymer thin film scaffolding.

    PubMed

    Komiyama, Hideaki; Iyoda, Tomokazu; Sanji, Takanobu

    2015-10-02

    We present an alternative approach to fabricating hexagonally arranged nanodot arrays of various metals by seed-mediated electroless plating with a cylinder-forming block copolymer thin film, PEO-b-PMA(Az), as a scaffold. Metal ions were selectively incorporated into PEO cylinders, followed by their reduction to metal and the etching of the scaffold to obtain highly ordered seed arrays of Au, Pd, and Pt. Nanodot arrays of the target metals (Au, Ag, and Ni) were selectively grown on the seed with their highly ordered arrangement by electroless plating. We studied the fabrication processes' suitability for control of the nanodot array size, as well as the plasmonic properties thereof.

  9. Metal nanodot arrays fabricated via seed-mediated electroless plating with block copolymer thin film scaffolding

    NASA Astrophysics Data System (ADS)

    Komiyama, Hideaki; Iyoda, Tomokazu; Sanji, Takanobu

    2015-10-01

    We present an alternative approach to fabricating hexagonally arranged nanodot arrays of various metals by seed-mediated electroless plating with a cylinder-forming block copolymer thin film, PEO-b-PMA(Az), as a scaffold. Metal ions were selectively incorporated into PEO cylinders, followed by their reduction to metal and the etching of the scaffold to obtain highly ordered seed arrays of Au, Pd, and Pt. Nanodot arrays of the target metals (Au, Ag, and Ni) were selectively grown on the seed with their highly ordered arrangement by electroless plating. We studied the fabrication processes’ suitability for control of the nanodot array size, as well as the plasmonic properties thereof.

  10. Metal removal and associated binding fraction transformation in contaminated river sediment washed by different types of agents

    PubMed Central

    Wang, Hong; Liu, Tongzhou; Feng, Shuai; Zhang, Weihua

    2017-01-01

    In ex-situ washing, HCl, EDTA and H2O2 solutions can effectively extract heavy metals in river sediment. Nevertheless they often target different sediment components, possibly transforming metal species into more bioavailable and hence toxic ones. This study, in batch settings, investigated the influences of different types of washing agents (i.e. HCl, EDTA and H2O2) on metal (i.e. Cu and Zn) removal from contaminated river sediment, destroy or dissolution of sediment components, and transformation of metal fractions during chemical washing treatment. Additionally, bioavailability of these metals left in the washed sediment was assessed. Results showed that HCl obtained the highest Cu and Zn removal through destroying the reducible, oxidizable and residual sediment components. Meanwhile, it transformed metal fractions to acid extractable one, resulting in an increase in metal bioavailability. Thus, the feasibility of washing with HCl for sediment remediation shall be reconsidered due to the caused high metal bioavailability. EDTA was capable of removing metals via direct complexation of labile metal species and indirect dissolution of reducible and oxidizable sediment components, where the transformation of corresponding metal binding fraction may occur. H2O2 obtained the lowest total Cu and Zn removal, but it preferentially removed the oxidizable metal species by oxidizing sulfides in the sediment. The bioavailable levels of Cu and Zn in the sediment washed by EDTA or H2O2 seemed not increase. To maintain a good balance between labile metal species removal and avoiding increase of metal bioavailability, EDTA and H2O2 are promising additives for metal removal by sediment washing. PMID:28350832

  11. Metal removal and associated binding fraction transformation in contaminated river sediment washed by different types of agents.

    PubMed

    Wang, Hong; Liu, Tongzhou; Feng, Shuai; Zhang, Weihua

    2017-01-01

    In ex-situ washing, HCl, EDTA and H2O2 solutions can effectively extract heavy metals in river sediment. Nevertheless they often target different sediment components, possibly transforming metal species into more bioavailable and hence toxic ones. This study, in batch settings, investigated the influences of different types of washing agents (i.e. HCl, EDTA and H2O2) on metal (i.e. Cu and Zn) removal from contaminated river sediment, destroy or dissolution of sediment components, and transformation of metal fractions during chemical washing treatment. Additionally, bioavailability of these metals left in the washed sediment was assessed. Results showed that HCl obtained the highest Cu and Zn removal through destroying the reducible, oxidizable and residual sediment components. Meanwhile, it transformed metal fractions to acid extractable one, resulting in an increase in metal bioavailability. Thus, the feasibility of washing with HCl for sediment remediation shall be reconsidered due to the caused high metal bioavailability. EDTA was capable of removing metals via direct complexation of labile metal species and indirect dissolution of reducible and oxidizable sediment components, where the transformation of corresponding metal binding fraction may occur. H2O2 obtained the lowest total Cu and Zn removal, but it preferentially removed the oxidizable metal species by oxidizing sulfides in the sediment. The bioavailable levels of Cu and Zn in the sediment washed by EDTA or H2O2 seemed not increase. To maintain a good balance between labile metal species removal and avoiding increase of metal bioavailability, EDTA and H2O2 are promising additives for metal removal by sediment washing.

  12. An improved Agrobacterium-mediated transformation system for the functional genetic analysis of Penicillium marneffei.

    PubMed

    Kummasook, Aksarakorn; Cooper, Chester R; Vanittanakom, Nongnuch

    2010-12-01

    We have developed an improved Agrobacterium-mediated transformation (AMT) system for the functional genetic analysis of Penicillium marneffei, a thermally dimorphic, human pathogenic fungus. Our AMT protocol included the use of conidia or pre-germinated conidia of P. marneffei as the host recipient for T-DNA from Agrobacterium tumefaciens and co-cultivation at 28°C for 36 hours. Bleomycin-resistant transformants were selected as yeast-like colonies following incubation at 37°C. The efficiency of transformation was approximately 123 ± 3.27 and 239 ± 13.12 transformants per plate when using 5 × 10(4) conidia and pre-germinated conidia as starting materials, respectively. Southern blot analysis demonstrated that 95% of transformants contained single copies of T-DNA. Inverse PCR was employed for identifying flanking sequences at the T-DNA insertion sites. Analysis of these sequences indicated that integration occurred as random recombination events. Among the mutants isolated were previously described stuA and gasC defective strains. These AMT-derived mutants possessed single T-DNA integrations within their particular coding sequences. In addition, other morphological and pigmentation mutants possessing a variety of gene-specific defects were isolated, including two mutants having T-DNA integrations within putative promoter regions. One of the latter integration events was accompanied by the deletion of the entire corresponding gene. Collectively, these results indicated that AMT could be used for large-scale, functional genetic analyses in P. marneffei. Such analyses can potentially facilitate the identification of those genetic elements related to morphogenesis, as well as pathogenesis in this medically important fungus.

  13. Prevention of KLF4-mediated tumor initiation and malignant transformation by UAB30 rexinoid

    PubMed Central

    Jiang, Wen; Deng, Wentao; Bailey, Sarah K.; Nail, Clint D.; Frost, Andra R.; Brouillette, Wayne J.; Muccio, Donald D.; Grubbs, Clinton J.; Ruppert, J. Michael; Lobo-Ruppert, Susan M.

    2009-01-01

    The transcription factor KLF4 acts in post-mitotic epithelial cells to promote differentiation, and functions in a context-dependent fashion as an oncogene. In the skin KLF4 is co-expressed with the nuclear receptors RARγ and RXRα, and formation of the skin permeability barrier is a shared function of these three proteins. We utilized a KLF4-transgenic mouse model of skin cancer in combination with cultured epithelial cells to examine functional interactions between KLF4 and retinoic acid receptors. In cultured cells, activation of a conditional, KLF4-estrogen receptor fusion protein by 4-hydroxytamoxifen resulted in rapid upregulation of transcripts for nuclear receptors including RARγ and RXRα. We tested retinoids in epithelial cell transformation assays, including an RAR-selective agonist (all-trans RA), an RXR-selective agonist (9-cis UAB30, rexinoid), and a pan agonist (9-cis RA). Unlike for several other genes, transformation by KLF4 was inhibited by each retinoid, implicating distinct nuclear receptor heterodimers as modulators of KLF4 transforming activity. When RXRα expression was suppressed by RNAi in cultured cells, transformation was promoted and the inhibitory effect of 9-cis UAB30 was attenuated. Similarly as shown for other mouse models of skin cancer, rexinoid prevented skin tumor initiation resulting from induction of KLF4 in basal keratinocytes. Rexinoid permitted KLF4 expression and KLF4-induced cell cycling, but attenuated the KLF4-induced misexpression of cytokeratin 1 in basal cells. Neoplastic lesions including hyperplasia, dysplasia and squamous cell carcinoma-like lesions were prevented for up to 30 days. Taken together, the results identify retinoid receptors including RXRα as ligand-dependent inhibitors of KLF4-mediated transformation or tumorigenesis. PMID:19197145

  14. Agrobacterium tumefaciens-mediated genetic transformation of a recalcitrant grain legume, lentil (Lens culinaris Medik).

    PubMed

    Akcay, Ufuk Celikkol; Mahmoudian, M; Kamci, H; Yucel, M; Oktem, H A

    2009-03-01

    A simple and reproducible Agrobacterium-mediated transformation protocol for a recalcitrant legume plant, lentil (Lens culinaris M.) is reported. Application of wounding treatments and efficiencies of three Agrobacterium tumefaciens strains, EHA105, C58C1, and KYRT1 were compared for T-DNA delivery into lentil cotyledonary node tissues. KYRT1 was found to be on average 2.8-fold more efficient than both EHA105 and C58C1 for producing transient beta-glucuronidase (GUS) gene (gus) expression on cotyledonary petioles. Wounding of the explants, use of an optimized transformation protocol with the application of acetosyringone and vacuum infiltration treatments in addition to the application of a gradually intensifying selection regime played significant roles in enhancing transformation frequency. Lentil explants were transformed by inoculation with Agrobacterium tumefaciens strain, KYRT1 harboring a binary vector pTJK136 that carried neomycin phosphotransferase gene (npt-II) and an intron containing gusA gene on its T-DNA region. GUS-positive shoots were micrografted on lentil rootstocks. Transgenic lentil plants were produced with an overall transformation frequency of 2.3%. The presence of the transgene in the lentil genome was confirmed by GUS assay, PCR, RT-PCR and Southern hybridization. The transgenic shoots grafted on rootstocks were successfully transferred to soil and grown to maturity in the greenhouse. GUS activity was detected in vegetative and reproductive organs of T(0), T(1), T(2) and T(3) plants. PCR assays of T(1), T(2) and T(3) progenies confirmed the stable transmission of the transgene to the next generations.

  15. Prevention of KLF4-mediated tumor initiation and malignant transformation by UAB30 rexinoid.

    PubMed

    Jiang, Wen; Deng, Wentao; Bailey, Sarah K; Nail, Clint D; Frost, Andra R; Brouillette, Wayne J; Muccio, Donald D; Grubbs, Clinton J; Ruppert, J Michael; Lobo-Ruppert, Susan M

    2009-02-01

    The transcription factor KLF4 acts in post-mitotic epithelial cells to promote differentiation and functions in a context-dependent fashion as an oncogene. In the skin KLF4 is co-expressed with the nuclear receptors RARgamma and RXRalpha, and formation of the skin permeability barrier is a shared function of these three proteins. We utilized a KLF4-transgenic mouse model of skin cancer in combination with cultured epithelial cells to examine functional interactions between KLF4 and retinoic acid receptors. In cultured cells, activation of a conditional, KLF4-estrogen receptor fusion protein by 4-hydroxytamoxifen resulted in rapid upregulation of transcripts for nuclear receptors including RARgamma and RXRalpha. We tested retinoids in epithelial cell transformation assays, including an RAR-selective agonist (all-trans RA), an RXR-selective agonist (9-cis UAB30, rexinoid), and a pan agonist (9-cis RA). Unlike for several other genes, transformation by KLF4 was inhibited by each retinoid, implicating distinct nuclear receptor heterodimers as modulators of KLF4 transforming activity. When RXRalpha expression was suppressed by RNAi in cultured cells, transformation was promoted and the inhibitory effect of 9-cis UAB30 was attenuated. Similarly as shown for other mouse models of skin cancer, rexinoid prevented skin tumor initiation resulting from induction of KLF4 in basal keratinocytes. Rexinoid permitted KLF4 expression and KLF4-induced cell cycling, but attenuated the KLF4-induced misexpression of cytokeratin 1 in basal cells. Neoplastic lesions including hyperplasia, dysplasia and squamous cell carcinoma-like lesions were prevented for up to 30 days. Taken together, the results identify retinoid receptors including RXRalpha as ligand-dependent inhibitors of KLF4-mediated transformation or tumorigenesis.

  16. Formation of nanostructured Group IIA metal activated sensors: The transformation of Group IIA metal compound sites

    NASA Astrophysics Data System (ADS)

    Tune, Travis C.; Baker, Caitlin; Hardy, Neil; Lin, Arthur; Widing, Timothy J.; Gole, James L.

    2015-05-01

    Trends in the Group IIA metal oxides and hydroxides of magnesium, calcium, and barium are unique in the periodic table. In this study we find that they display novel trends as decorating nanostructures for extrinsic semiconductor interfaces. The Group IIA metal ions are strong Lewis acids. We form these M2+ ions in aqueous solution and bring these solutions in contact with a porous silicon interface to form interfaces for conductometric measurements. Observed responses are consistent with the formation of MgO whereas the heavier elements display behaviors which suggest the effect of their more basic nature. Mg(OH)2, when formed, represents a weak base whereas the heavier metal hydroxides of Ca, Sr, and Ba are strong bases. However, the hydroxides tend to give up hydrogen and act as Brönsted acids. For the latter elements, the reversible interaction response of nanostructures deposited to the porous silicon (PS) interface is modified, as the formation of more basic sites appears to compete with M2+ Lewis acidity and hydroxide Brönsted acidity. Mg2+ forms an interface whose response to the analytes NH3 and NO is consistent with MgO and well explained by the recently developing Inverse Hard/Soft Acid/Base model. The behavior of the Ca2+ and Ba2+ decorated interfaces as they interact with the hard base NH3 follows a reversal of the model, indicating a decrease in acidic character as the observed conductometric response suggests the interaction with hydroxyl groups. A change from oxide-like to hydroxide-like constituents is supported by XPS studies. The changes in conductometric response is easily monitored in contrast to changes associated with the Group IIA oxides and hydroxides observed in XPS, EDAX, IR, and NMR measurements.

  17. Recovery of transgenic plants by pollen-mediated transformation in Brassica juncea.

    PubMed

    Wang, Jingxue; Li, Yonghu; Liang, Chao

    2008-06-01

    The aroA-M1 encoding the mutant of 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPS) was introduced into the Brassica juncea genome by sonication-assisted, pollen-mediated transformation. The plasmid DNA and collected pollen grains were mixed in 0.3 mol/L sucrose solution and treated with mild ultrasonication. The treated pollen was then pollinated onto the oilseed stigmas after the stamens were removed artificially. Putative transgenic plants were obtained by screening germinating seeds on a medium containing glyphosate. Southern blot analysis of glyphosate-resistant plants indicated that the aroA-M1 gene had been integrated into the oilseed genome. Western blot analysis further confirmed that the EPSPS coded by aroA-M1 gene was expressed in transgenic plants. The transgenic plants exhibited increased resistance to glyphosate compared to untransformed plants. Some of those transgenic plants had considerably high resistance to glyphosate. The genetic analysis of T1 progeny further confirmed that the inheritance of the introduced genes followed the Mendelian rules. The results indicated that foreign genes can be transferred by pollen-mediated transformation combined with mild ultrasonication.

  18. Influence of organic matter transformations on the bioavailability of heavy metals in a sludge based compost.

    PubMed

    Ingelmo, Florencio; Molina, Maria José; Soriano, Maria Desamparados; Gallardo, Antonio; Lapeña, Leonor

    2012-03-01

    The agricultural use of anaerobically digested sewage sludge (ADSS) as stable, mature compost implies knowing its total content in heavy metals and their bioavailability. This depends not only on the initial characteristics of the composted substrates but also on the organic matter transformations during composting which may influence the chemical form of the metals and their bioavailability. The objective of this work was to examine the relationships between the changes in the organic matter content and humus fractions, and the bioavailability of heavy metals. A detailed sampling at 0, 14, 84, and 140 days of the composting process was performed to measure C contents in humic acids (HAs), fulvic acids, (FAs) and humin, the total content of Zn, Pb, Cu, Ni, and Cd, and also their distribution into mobile and mobilisable (MB), and low bioavailability (LB) forms. Significant changes of C contents in HA, FA, and Humin, and in the FA/HA, HA/Humin and C(humus)/TOC ratios were observed during composting. The MB and LB fractions of each metal also varied significantly during composting. The MB fraction increased for Zn, Cu, Ni, and Cd, and the LB fraction increased for Pb. Stepwise linear regressions and quadratic curve estimation conducted on the MB and LB fractions of each metal as dependent on the measured organic variables suggested that Zn bioavailability was mainly associated to percentage of C in FAs. Bioavailability of Cu, Ni and Cd during composting was associated to humin and HAs. Pb concentration increased in the LB form, and its variations followed a quadratic function with the C(humus)/TOC ratio. Our results suggest that the composting process renders the metals in more available forms. The main forms of metal binding in the sludge and their availability in the final compost may be better described when metal fractionation obtained in sequential extraction and humus fractionation during composting are considered together.

  19. Extracellular Saccharide-Mediated Reduction of Au(3+) to Gold Nanoparticles: New Insights for Heavy Metals Biomineralization on Microbial Surfaces.

    PubMed

    Kang, Fuxing; Qu, Xiaolei; Alvarez, Pedro J J; Zhu, Dongqiang

    2017-02-15

    Biomineralization is a critical process controlling the biogeochemical cycling, fate, and potential environmental impacts of heavy metals. Despite the indispensability of extracellular polymeric substances (EPS) to microbial life and their ubiquity in soil and aquatic environments, the role played by EPS in the transformation and biomineralization of heavy metals is not well understood. Here, we used gold ion (Au(3+)) as a model heavy metal ion to quantitatively assess the role of EPS in biomineralization and discern the responsible functional groups. Integrated spectroscopic analyses showed that Au(3+)was readily reduced to zerovalent gold nanoparticles (AuNPs, 2-15 nm in size) in aqueous suspension of Escherichia coli or dissolved EPS extracted from microbes. The majority of AuNPs (95.2%) was formed outside Escherichia coli cells, and the removal of EPS attached to cells pronouncedly suppressed Au(3+) reduction, reflecting the predominance of the extracellular matrix in Au(3+) reduction. XPS, UV-vis, and FTIR analyses corroborated that Au(3+) reduction was mediated by the hemiacetal groups (aldehyde equivalents) of reducing saccharides of EPS. Consistently, the kinetics of AuNP formation obeyed pseudo-second-order reaction kinetics with respect to the concentrations of Au(3+) and the hemiacetal groups in EPS, with minimal dependency on the source of microbial EPS. Our findings indicate a previously overlooked, universally significant contribution of EPS to the reduction, mineralization, and potential detoxification of metal species with high oxidation state.

  20. AgarTrap: a simplified Agrobacterium-mediated transformation method for sporelings of the liverwort Marchantia polymorpha L.

    PubMed

    Tsuboyama, Shoko; Kodama, Yutaka

    2014-01-01

    The liverwort Marchantia polymorpha L. is being developed as an emerging model plant, and several transformation techniques were recently reported. Examples are biolistic- and Agrobacterium-mediated transformation methods. Here, we report a simplified method for Agrobacterium-mediated transformation of sporelings, and it is termed Agar-utilized Transformation with Pouring Solutions (AgarTrap). The procedure of the AgarTrap was carried out by simply exchanging appropriate solutions in a Petri dish, and completed within a week, successfully yielding sufficient numbers of independent transformants for molecular analysis (e.g. characterization of gene/protein function) in a single experiment. The AgarTrap method will promote future molecular biological study in M. polymorpha.

  1. Transformation and availability of nutrients and heavy metals during integrated composting-vermicomposting of sewage sludges.

    PubMed

    Hait, Subrata; Tare, Vinod

    2012-05-01

    Transformation and availability of nutrients and some heavy metals were assessed during the integrated composting-vermicomposting of both primary sewage sludge (PSS) and waste activated sewage sludge (WAS) using matured vermicompost as indigenous bulking material and employing Eisenia fetida as earthworm species. Vermicomposting resulted in significant increase in total N (TN) (PSS: 41.7-64.6%, F=11.6, P<0.05; WAS: 36.4-58.6%, F=6.4, P<0.05), water soluble N (WSN) (PSS: 37.1-50.5%, F=30.1, P<0.05; WAS: 40.1-53.0%, F=27.6, P<0.05), total P (TP) (PSS: 39.9-69.8%, F=27.1, P<0.05; WAS: 32.2-56.6%, F=21.4, P<0.05) and water soluble P (WSP) (PSS: 25.2-34.3%, F=163.9, P<0.05; WAS: 24.1-34.2%, F=144.3, P<0.05) as compared to the initial compost material depending on different experimental conditions. The study demonstrated that the vermicomposting significantly improved the availability of nutrients in sewage sludges. In addition, vermicomposting considerably reduced the availability of heavy metals except Fe and Mn, presumably by forming organic-bound complexes in spite of several fold increase in their total content. The environmental conditions (i.e., temperature and relative humidity), in general, showed significant effect on the transformation and availability of nutrients and heavy metals. There was no effect of earthworm density on the transformation and availability of heavy metals and nutrients except N and P, possibly due to prior exposure during acclimation period in sewage sludge. Copyright © 2012 Elsevier Inc. All rights reserved.

  2. Transformation of triclosan by laccase catalyzed oxidation: The influence of humic acid-metal binding process.

    PubMed

    Lu, Junhe; Shi, Yuanyuan; Ji, Yuefei; Kong, Deyang; Huang, Qingguo

    2017-01-01

    Laccase is a widely present extracellular phenoloxidase excreted by fungi, bacteria, and high plants. It is able to catalyze one-electron oxidation of phenolic compounds into radical intermediates that can subsequently couple to each other via covalent bonds. These reactions are believed to play an important role in humification process and the transformation of contaminants containing phenolic functionalities in the environment. In this study, we investigated the kinetics of triclosan transformation catalyzed by laccase. It was found that the rate of triclosan oxidation was first order to the concentrations of both substrate and enzyme. Humic acid (HA) could inhibit the reaction by quenching the radical intermediate of triclosan generated by laccase oxidation. Such inhibition was more significant in the presence of divalent metal cations. This is because that binding to metal ions neutralized the negative charge of HA molecules, thus making them more accessible to laccase molecule that is also negatively charged. Therefore, it has greater chance to quench the radical intermediate that is very unstable and can only diffuse a limited distance after being released from the enzyme catalytic center. Based on these understandings, a reaction model was developed by integration of metal-HA binding equilibriums and kinetic equations. This model precisely predicted the transformation rate of triclosan in the presence of HA and divalent metal ions including Ca(2+), Mg(2+), Cd(2+), Co(2+), Mn(2+), Ba(2+), and Zn(2+). Overall, this work reveals important insights into laccase catalyzed oxidative coupling process. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Efficient gene targeting in Penicillium chrysogenum using novel Agrobacterium-mediated transformation approaches.

    PubMed

    de Boer, Paulo; Bronkhof, Jurian; Dukiќ, Karolina; Kerkman, Richard; Touw, Hesselien; van den Berg, Marco; Offringa, Remko

    2013-12-01

    The industrial production of β-lactam antibiotics by Penicillium chrysogenum has increased tremendously over the last decades, however, further optimization via classical strain and process improvement has reached its limits. The availability of the genome sequence provides new opportunities for directed strain improvement, but this requires the establishment of an efficient gene targeting (GT) system. Recently, mutations affecting the non-homologous end joining (NHEJ) pathway were shown to increase GT efficiencies following PEG-mediated DNA transfer in P. chrysogenum from 1% to 50%. Apart from direct DNA transfer many fungi can efficiently be transformed using the T-DNA transfer system of the soil bacterium Agrobacterium tumefaciens, however, for P. chrysogenum no robust system for Agrobacterium-mediated transformation was available. We obtained efficient AMT of P. chrysogenum spores with the nourseothricin acetyltransferase gene as selection marker, and using this system we investigated if AMT in a NHEJ mutant background could further enhance GT efficiencies. In general, AMT resulted in higher GT efficiencies than direct DNA transfer, although the final frequencies depended on the Agrobacterium strain and plasmid backbone used. Providing overlapping and complementing fragments on two different plasmid backbones via the same Agrobacterium host was shown to be most effective. This so-called split-marker or bi-partite method resulted in highly efficient GT (>97%) almost exclusively without additional ectopic T-DNA insertions. As this method provides for an efficient GT method independent of protoplasts, it can be applied to other fungi for which no protoplasts can be generated or for which protoplast transformation leads to varying results.

  4. Delineation of First-Order Elastic Property Closures for Hexagonal Metals Using Fast Fourier Transforms

    PubMed Central

    Landry, Nicholas W.; Knezevic, Marko

    2015-01-01

    Property closures are envelopes representing the complete set of theoretically feasible macroscopic property combinations for a given material system. In this paper, we present a computational procedure based on fast Fourier transforms (FFTs) for delineation of elastic property closures for hexagonal close packed (HCP) metals. The procedure consists of building a database of non-zero Fourier transforms for each component of the elastic stiffness tensor, calculating the Fourier transforms of orientation distribution functions (ODFs), and calculating the ODF-to-elastic property bounds in the Fourier space. In earlier studies, HCP closures were computed using the generalized spherical harmonics (GSH) representation and an assumption of orthotropic sample symmetry; here, the FFT approach allowed us to successfully calculate the closures for a range of HCP metals without invoking any sample symmetry assumption. The methodology presented here facilitates for the first time computation of property closures involving normal-shear coupling stiffness coefficients. We found that the representation of these property linkages using FFTs need more terms compared to GSH representations. However, the use of FFT representations reduces the computational time involved in producing the property closures due to the use of fast FFT algorithms. Moreover, FFT algorithms are readily available as opposed to GSH codes. PMID:28793566

  5. Delineation of First-Order Elastic Property Closures for Hexagonal Metals Using Fast Fourier Transforms.

    PubMed

    Landry, Nicholas W; Knezevic, Marko

    2015-09-17

    Property closures are envelopes representing the complete set of theoretically feasible macroscopic property combinations for a given material system. In this paper, we present a computational procedure based on fast Fourier transforms (FFTs) to delineation of elastic property closures for hexagonal close packed (HCP) metals. The procedure consists of building a database of non-zero Fourier transforms for each component of the elastic stiffness tensor, calculating the Fourier transforms of orientation distribution functions (ODFs), and calculating the ODF-to-elastic property bounds in the Fourier space. In earlier studies, HCP closures were computed using the generalized spherical harmonics (GSH) representation and an assumption of orthotropic sample symmetry; here, the FFT approach allowed us to successfully calculate the closures for a range of HCP metals without invoking any sample symmetry assumption. The methodology presented here facilitates for the first time computation of property closures involving normal-shear coupling stiffness coefficients. We found that the representation of these property linkages using FFTs need more terms compared to GSH representations. However, the use of FFT representations reduces the computational time involved in producing the property closures due to the use of fast FFT algorithms. Moreover, FFT algorithms are readily available as opposed to GSH codes.

  6. Prolate rotation and metallicity gradient in the transforming dwarf galaxy Phoenix

    NASA Astrophysics Data System (ADS)

    Kacharov, Nikolay; Battaglia, Giuseppina; Rejkuba, Marina; Cole, Andrew A.; Carrera, Ricardo; Fraternali, Filippo; Wilkinson, Mark I.; Gallart, Carme G.; Irwin, Mike; Tolstoy, Eline

    2017-04-01

    Transition type dwarf galaxies are thought to be systems undergoing the process of transformation from a star-forming into a passively evolving dwarf, which makes them particularly suitable to study evolutionary processes driving the existence of different dwarf morphological types. Here we present results from a spectroscopic survey of ∼200 individual red giant branch stars in the Phoenix dwarf, the closest transition type with a comparable luminosity to 'classical' dwarf galaxies. We measure a systemic heliocentric velocity Vhelio = -21.2 ± 1.0 km s-1. Our survey reveals the clear presence of prolate rotation that is aligned with the peculiar spatial distribution of the youngest stars in Phoenix. We speculate that both features might have arisen from the same event, possibly an accretion of a smaller system. The evolved stellar population of Phoenix is relatively metal-poor (<[Fe/H] > = -1.49 ± 0.04 dex) and shows a large metallicity spread (σ[Fe/H] = 0.51 ± 0.04 dex), with a pronounced metallicity gradient of -0.13 ± 0.01 dex arcmin-1 similar to luminous, passive dwarf galaxies. We also report a discovery of an extremely metal-poor star candidate in Phoenix and discuss the importance of correcting for spatial sampling when interpreting the chemical properties of galaxies with metallicity gradients. This study presents a major leap forward in our knowledge of the internal kinematics of the Phoenix transition type dwarf galaxy and the first wide area spectroscopic survey of its metallicity properties. A table containing the measured velocities, metallicities, and CaT equivalent widths of all spectroscopic targets is available online at the CDS.

  7. Phagocytosis and transforming activity of crystalline metal sulfide particles are related to their negative surface charge

    SciTech Connect

    Abbracchio, M.P.; Heck, J.D.; Costa, M.

    1982-01-01

    Crystalline nickel sulfide (alpha NiS) and cobalt sulfide (CoS2) particles can cause greater cell transformation and cellular toxicity than the respective amorphous metal sulfide particles. Cultured mammalian cells phagocytose the crystalline metal sulfide particles more readily than the amorphous ones. In the case of the nickel sulfides, the crystalline metal sulfide particles had negatively charged surfaces (Zeta potential: -27.012 mV) in contrast to the amorphous particles, which were positively charge (Zeta potential: +9.174 mV). X-ray photoelectron spectroscopy analysis of amorphous and crystalline NiS particles revealed that the outermost surface (1-4 nm) of the two particles had striking differences in Ni/S ratios and in their sulfur oxidation states. Rendering particles' surfaces more negative by reduction with lithium aluminum hydride enhanced their phagocytosis, and in the case of amorphous NiS chemical reduction resulted in an incidence of morphological transformation of Syrian hamster embryo cells comparable to that observed with untreated crystalline alpha NiS.

  8. High frequency regeneration via direct somatic embryogenesis and efficient Agrobacterium- mediated genetic transformation of tobacco

    PubMed Central

    Pathi, Krishna Mohan; Tula, Suresh; Tuteja, Narendra

    2013-01-01

    A direct somatic embryogenesis protocol was developed for four cultivars of Nicotiana species, by using leaf disc as an explant. Direct somatic embryogenesis of Nicotiana by using BAP and IAA has not been investigated so far. This method does not require formation of callus tissues which leads to somaclonal variations. The frequency of somatic embryogenesis was strongly influenced by the plant growth hormones. The somatic embryos developing directly from explant tissue were noticed after 6 d of culture. Somatic embryogenesis of a high frequency (87–96%) was observed in cultures of the all four genotypes (Nicotiana tabacum, N. benthamiyana, N. xanthi, N. t cv petihavana). The results showed that the best medium for direct somatic embryogenesis was MS supplemented with 2.5 mg/l, 0.2 mg/l IAA and 2% sucrose. Subculture of somatic embryos onto hormone free MS medium resulted in their conversion into plants for all genotypes. About 95% of the regenerated somatic embryos germinated into complete plantlets. The plants showed morphological and growth characteristics similar to those of seed-derived plants. Explants were transformed using Agrobacterium tumifacious LBA4404 plasmid pCAMBIA1301 harboring the GUS gene. The regenerated transgenic plants were confirmed by PCR analysis and histochemical GUS assay. The transformation efficiency obtained by using the Agrobacterium- mediated transformation was more than 95%. This method takes 6 wk to accomplish complete transgenic plants through direct somatic embryogenesis. The transgenic plantlets were acclimatized successfully with 98% survival in greenhouse and they showed normal morphological characteristics and were fertile. The regeneration and transformation method described herein is very simple, highly efficient and fast for the introduction of any foreign gene directly in tobacco through direct somatic embryogenesis. PMID:23518589

  9. Agrobacterium tumefaciens-mediated sorghum transformation using a mannose selection system.

    PubMed

    Gao, Zhensheng; Xie, Xueju; Ling, Yan; Muthukrishnan, Subbarat; Liang, George H

    2005-11-01

    A dual-marker plasmid containing the selectable marker gene, manA, and the reporter gene, sgfp, was used to transform immature sorghum embryos by employing an Agrobacterium-mediated system. Both genes were under the control of the ubi1 promoter in a binary vector pPZP201. The Escherichia coli phosphomannose isomerase (PMI) gene, pmi, was used as the selectable marker gene and mannose was used as the selective agent. The sgfp gene encoding green fluorescence protein (GFP) was the reporter gene and served as a visual screening marker. A total of 167 transgenic plants were obtained from nine different embryogenic callus lines grown on a selection medium containing 1%-2% mannose. Embryoids and shoots regenerated via embryogenesis, that showed strong GFP fluorescence, were selected from two sorghum genotypes: C401, an inbred line, and Pioneer 8505, a commercial hybrid. The GFP accumulation in transgenic plants was observed with a dissecting stereomicroscope. The integration and expression of the manA gene was confirmed by Southern blot and Western blot analyses, and the feasibility of manA selection was demonstrated by the chlorophenol red (CPR) assay. Our results indicated that transgenes segregated in the Mendelian fashion in the T1 generation. The conversion of mannose to a metabolizable fructose carbon source is beneficial to plants. In addition, except in soybean and a few legumes, no endogenous PMI activity has been detected in plant species, indicating that PMI is useful in the transformation of sorghum. In addition, PMI has no sequence homology to known allergens. Optimization of this selection system for sorghum transformation provides an efficient way to produce transgenic plants without using antibiotic or herbicidal agents as selectable markers, and our results showed that the transformation efficiency reached 2.88% for Pioneer 8505 and 3.30% for C401, both values higher than in previously published reports.

  10. Interaction of metals with peroxidase--mediated luminol-enhanced, chemiluminescence (PLmCL).

    PubMed

    Coteur, G; Dubois, P

    2004-01-01

    The peroxidase-mediated luminol-enhanced chemiluminescence (PLmCL) method has been used to study the in vitro effect of contaminants such as heavy metals on the reactive oxygen species production by immunocytes. We were interested to know whether metals could directly affect peroxidase-mediated luminescence, taking horseradish peroxidase (HRP) as a model enzyme, since this could contribute to the inhibition of immunocyte LmCL. Copper inhibited PLmCL in a dose-dependent manner, while cadmium, iron, silver and lead only partly decreased the signal in the concentration range tested. In contrast, zinc enhanced the signal at high concentrations. Eventually, chromium, mercury and aluminium did not affect PLmCL. It is suggested that these effects reflect the ability of the metals to interact with the active site of the peroxidase. These results demonstrate that such interactions have to be considered when interpreting the effects of metals on immunocytes using the LmCL method.

  11. Agrobacterium-mediated genetic transformation of Coffea arabica (L.) is greatly enhanced by using established embryogenic callus cultures

    PubMed Central

    2011-01-01

    Background Following genome sequencing of crop plants, one of the main challenges today is determining the function of all the predicted genes. When gene validation approaches are used for woody species, the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. Embryogenic calli have frequently been the target tissue for transformation, but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants, including Coffea arabica. Results We identified the conditions required for successful long-term proliferation of embryogenic cultures in C. arabica and designed a highly efficient and reliable Agrobacterium tumefaciens-mediated transformation method based on these conditions. The transformation protocol with LBA1119 harboring pBin 35S GFP was established by evaluating the effect of different parameters on transformation efficiency by GFP detection. Using embryogenic callus cultures, co-cultivation with LBA1119 OD600 = 0.6 for five days at 20 °C enabled reproducible transformation. The maintenance conditions for the embryogenic callus cultures, particularly a high auxin to cytokinin ratio, the age of the culture (optimum for 7-10 months of proliferation) and the use of a yellow callus phenotype, were the most important factors for achieving highly efficient transformation (> 90%). At the histological level, successful transformation was related to the number of proembryogenic masses present. All the selected plants were proved to be transformed by PCR and Southern blot hybridization. Conclusion Most progress in increasing transformation efficiency in coffee has been achieved by optimizing the production conditions of embryogenic cultures used as target tissues for transformation. This is the first time that a strong positive effect of the age of the culture on transformation efficiency was demonstrated. Our results make Agrobacterium-mediated

  12. Characterization of Solid State Phase Transformation in Continuously Heated and Cooled Ferritic Weld Metal

    SciTech Connect

    Narayana, B; Mills, Michael J.; Specht, Eliot D; Santella, Michael L; Babu, Sudarsanam Suresh

    2010-12-01

    Arc welding processes involve cooling rates that vary over a wide range (1-100 K/s). The final microstructire is thus a product of the heating and cooling cycles experienced by the weld in addition to the weld composition. It has been shown that the first phase to form under weld cooling conditions may not be that predicted by equilibrium calculations. The partitioning of different interstitial/substitutional alloying elements at high temperatures can dramatically affect the subsequent phase transformations. In order to understand the effect of alloying on phase transformation temperatures and final microstructures time-resolved X-ray diffraction technique has been successfully used for characterization. The work by Jacot and Rappaz on pearlitic steels provided insight into austenitization of hypoeutectic steels using a finite volume model. However there is very little work done on the effect of heating and cooling rates on the phase transformation paths in bainitic/martensitic steels and weld metals. Previous work on a weld with higher aluminum content, deposited with a FCAW-S process indicated that even at aluminum levels where the primary phase to solidify from liquid should be delta ferrite, non-equilibrium austenite was observed. The presence of inhomogeneity in composition of the parent microstructure has been attributed to differences in transformation modes, temperatures and microstructures in dual-phase, TRIP steels and ferritic welds. The objectives of the work included the identification of the stability regions of different phases during heating and cooling, differences in the effect of weld heating and cooling rates on the phase transformation temperatures, and the variation in phase fractions of austenite and ferrite in the two phase regions as a function of temperature. The base composition used for the present work is a Fe-1%Al-2%Mn-1%Ni-0.04%C weld metal. A pseudo-binary phase diagram shows the expected solidification path under equilibrium

  13. Plasmon-mediated Photoelectron Emission from Single, Supported Metal Nanoparticles

    NASA Astrophysics Data System (ADS)

    Grubisic, Andrej; Nesbitt, David

    2014-03-01

    Coherent multiphoton photoelectron emission (MPPE) from supported metal nanoparticles/structures has been studied at a single-particle level via scanning photoemission imaging microscopy (SPIM). Resonant excitation of localized surface plasmons (LSPs) with ultrafast laser pulses is shown to greatly amplify the photoelectron emission rate from metallic nanoparticles. In the limit of a large number of plasmon excitations, the n-photon photoelectron current scales rapidly with the electromagnetic near-field enhancement factor (|E|/|E0|)2n, indicating coherent MPPE as an extremely sensitive probe of the particle near-field. Additionally, our velocity map imaging (VMI) measurements of angle- and energy- resolved photoelectron distributions emitted from single plasmonic nanoparticles will be highlighted, with results shedding light into the complex dynamics of plasmon-induced photoelectron emission. We gratefully acknowledge Air Force Office of Scientific Research, National Science Foundation, and the National Institute of Standards and Technology for support of this work.

  14. Disclination mediated dynamic recrystallization in metals at low temperature

    NASA Astrophysics Data System (ADS)

    Aramfard, Mohammad; Deng, Chuang

    2015-09-01

    Recrystallization is one of the most important physical phenomena in condensed matter that has been utilized for materials processing for thousands of years in human history. It is generally believed that recrystallization is thermally activated and a minimum temperature must be achieved for the necessary atomic mechanisms to occur. Here, using atomistic simulations, we report a new mechanism of dynamic recrystallization that can operate at temperature as low as T = 10 K in metals during deformation. In contrast to previously proposed dislocation-based models, this mechanism relies on the generation of disclination quadrupoles, which are special defects that form during deformation when the grain boundary migration is restricted by structural defects such as triple junctions, cracks or obstacles. This mechanism offers an alternative explanation for the grain refinement in metals during severe plastic deformation at cryogenic temperature and may suggest a new method to tailor the microstructure in general crystalline materials.

  15. Disclination mediated dynamic recrystallization in metals at low temperature.

    PubMed

    Aramfard, Mohammad; Deng, Chuang

    2015-09-16

    Recrystallization is one of the most important physical phenomena in condensed matter that has been utilized for materials processing for thousands of years in human history. It is generally believed that recrystallization is thermally activated and a minimum temperature must be achieved for the necessary atomic mechanisms to occur. Here, using atomistic simulations, we report a new mechanism of dynamic recrystallization that can operate at temperature as low as T = 10 K in metals during deformation. In contrast to previously proposed dislocation-based models, this mechanism relies on the generation of disclination quadrupoles, which are special defects that form during deformation when the grain boundary migration is restricted by structural defects such as triple junctions, cracks or obstacles. This mechanism offers an alternative explanation for the grain refinement in metals during severe plastic deformation at cryogenic temperature and may suggest a new method to tailor the microstructure in general crystalline materials.

  16. Disclination mediated dynamic recrystallization in metals at low temperature

    PubMed Central

    Aramfard, Mohammad; Deng, Chuang

    2015-01-01

    Recrystallization is one of the most important physical phenomena in condensed matter that has been utilized for materials processing for thousands of years in human history. It is generally believed that recrystallization is thermally activated and a minimum temperature must be achieved for the necessary atomic mechanisms to occur. Here, using atomistic simulations, we report a new mechanism of dynamic recrystallization that can operate at temperature as low as T = 10 K in metals during deformation. In contrast to previously proposed dislocation-based models, this mechanism relies on the generation of disclination quadrupoles, which are special defects that form during deformation when the grain boundary migration is restricted by structural defects such as triple junctions, cracks or obstacles. This mechanism offers an alternative explanation for the grain refinement in metals during severe plastic deformation at cryogenic temperature and may suggest a new method to tailor the microstructure in general crystalline materials. PMID:26374603

  17. Extraordinary rates of transition metal ion-mediated ribozyme catalysis

    PubMed Central

    Roychowdhury-Saha, Manami; Burke, Donald H.

    2006-01-01

    In pre-steady-state, fast-quench kinetic analysis, the tertiary-stabilized hammerhead ribozyme “RzB” cleaves its substrate RNA with maximal measured k obs values of ∼3000 min−1 in 1 mM Mn2+ and ∼780 min−1 in 1 mM Mg2+ at 37°C (pH 7.4). Apparent pKa for the catalytic general base is ∼7.8–8.5, independent of the corresponding metal hydrate pKa, suggesting potential involvement of a nucleobase as general base as suggested previously from nucleobase substitution studies. The pH-rate profile is bell-shaped for Cd2+, for which the general catalytic acid has a pKa of 7.3 ± 0.1. Simulations of the pH-rate relation suggest a pKa for the general catalytic acid to be ∼9.5 in Mn2+ and >9.5 in Mg2+. The acid pKa's follow the trend in the pKa of the hydrated metal ions but are displaced by ∼1–2 pH units in the presence of Cd2+ and Mn2+. One possible explanation for this trend is direct metal ion coordination with a nucleobase, which then acts as general acid. PMID:16912216

  18. Purinergic signalling underlies transforming growth factor‐β‐mediated bladder afferent nerve hyperexcitability

    PubMed Central

    Gonzalez, Eric J.; Heppner, Thomas J.; Nelson, Mark T.

    2016-01-01

    Key points The sensory components of the urinary bladder are responsible for the transduction of bladder filling and are often impaired with neurological injury or disease.Elevated extracellular ATP contributes, in part, to bladder afferent nerve hyperexcitability during urinary bladder inflammation or irritation.Transforming growth factor‐β1 (TGF‐β1) may stimulate ATP release from the urothelium through vesicular exocytosis mechanisms with minimal contribution from pannexin‐1 channels to increase bladder afferent nerve discharge.Bladder afferent nerve hyperexcitability and urothelial ATP release with CYP‐induced cystitis is decreased with TGF‐β inhibition.These results establish a causal link between an inflammatory mediator, TGF‐β, and intrinsic signalling mechanisms of the urothelium that may contribute to the altered sensory processing of bladder filling. Abstract The afferent limb of the micturition reflex is often compromised following bladder injury, disease and inflammatory conditions. We have previously demonstrated that transforming growth factor‐β (TGF‐β) signalling contributes to increased voiding frequency and decreased bladder capacity with cystitis. Despite the functional presence of TGF‐β in bladder inflammation, the precise mechanisms of TGF‐β mediating bladder dysfunction are not yet known. Thus, the present studies investigated the sensory components of the urinary bladder that may underlie the pathophysiology of aberrant TGF‐β activation. We utilized bladder–pelvic nerve preparations to characterize bladder afferent nerve discharge and the mechanisms of urothelial ATP release with distention. Our findings indicate that bladder afferent nerve discharge is sensitive to elevated extracellular ATP during pathological conditions of urinary bladder inflammation or irritation. We determined that TGF‐β1 may increase bladder afferent nerve excitability by stimulating ATP release from the urothelium via vesicular

  19. Carcinogenic potential of metal nanoparticles in BALB/3T3 cell transformation assay.

    PubMed

    Sighinolfi, G L; Artoni, E; Gatti, A M; Corsi, L

    2016-05-01

    Metal-based nanoparticles (NPs), are currently used in many application fields including consumer products, pharmaceuticals, and biomedical treatments. In spite to their wide applications, an in-depth study of their potential toxic effects is still lacking. The aim of the present research was to investigate the potential initiator or promoter-like activity of different metallic NPs such as gold, iron, cobalt, and cerium using the Balb/3T3 two-stage transformation assay. The results indicated that all the selected metallic NPs, except for cobalt, when used as initiators did not induce any transformation in Balb/3T3 cell line. Moreover, Au and Fe3 O4 NPs, when used in place of the tumor promoter treatment TPA, increased significantly the number of Foci/dish as compared to the MCA treatment alone. The number of Foci/dish was 2.6 for Au NPs and 2.13 for Fe3 O4 ones, similar to those obtained by the positive control treatment (MCA + TPA), whereas 1.27 for MCA treatment alone. On the contrary, CeO2 NPs did not show any difference in the number of Foci/dish, as compared to MCA alone, but it decreased the number of foci by 65% in comparison to the positive control (MCA + TPA). As expected, cobalt NPs showed an increased cytotoxicity and only a few surviving cells were found at the time of analysis showing a number of Foci/dish of 0.13. For the first time, our data clearly showed that Au and Fe3 O4 NPs act as promoters in the two stage transformational assay, suggesting the importance to fully investigate the NPs carcinogenic potential with different models. © 2014 Wiley Periodicals, Inc.

  20. Effects of biochar on nitrogen transformation and heavy metals in sludge composting.

    PubMed

    Liu, Wei; Huo, Rong; Xu, Junxiang; Liang, Shuxuan; Li, Jijin; Zhao, Tongke; Wang, Shutao

    2017-07-01

    Composting is regarded as an effective treatment to suppress pathogenic organisms and stabilize the organic material in sewage sludge. This study investigated the use of biochar as an amendment to improve the composting effectiveness and reduce the bioavailability of heavy metals and loss of nitrogen during composting. Biochar of 0%, 1%, 3%, 5% and 7% were added into a mixture of sludge and straw, respectively. The use of biochar, even in small amounts, altered the composting process and the properties of the end products. Biochar addition resulted in a higher pile temperature (66°C) and could reduce nitrogen loss by transforming ammonium into nitrite. In the 5% biochar group, the final product from sludge composting, ammonia nitrogen, decreased by 22.4% compared to the control, and nitrate nitrogen increased by 310.6%. Considering temperature and N transformation, the treatment with 5% biochar is suggested for sludge composting. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Transformation of acoustic waves in periodic metal grating sandwiched between piezoelectric and dielectric.

    PubMed

    Naumenko, Natalya F; Abbott, Benjamin P

    2011-10-01

    The mechanism of SAW transformation with variation of film thickness is investigated in a piezoelectric substrate with a metal grating overlaid by a dielectric film, via simulation and visualization of the acoustic fields. By way of example, two orientations of lithium niobate substrates are analyzed, YX-LN and 128°YX-LN, with a Cu grating and an isotropic silica glass overlay. The motions, which follow the wave propagation in the sagittal plane, are visualized within two periods of the grating, with added contour plots showing the shear horizontal displacements. The continuous transformation of the wave's nature is investigated for each wave propagating in the analyzed material structures when the film thickness is increased from zero to a few wavelengths. The examples of the SAW transformation into boundary waves and into plate modes of different polarization have been found and investigated. The behavior of the SAW characteristics in the grating is correlated with transformation of the wave structure with increasing overlay thickness.

  2. Influence of phase transformation on stress evolution during growth of metal thin films on silicon.

    PubMed

    Fillon, A; Abadias, G; Michel, A; Jaouen, C; Villechaise, P

    2010-03-05

    In situ stress measurements during two-dimensional growth of low mobility metal films on amorphous Si were used to demonstrate the impact of interface reactivity and phase transformation on stress evolution. Using Mo1-xSix films as examples, the results show that the tensile stress rise, which develops after the film has become crystalline, is correlated with an increase in lateral grain size. The origin of the tensile stress is attributed to the volume change resulting from the alloy crystallization, which occurs at a concentration-dependent critical thickness.

  3. Transformation and Precipitation of Toxic Metals by ’Pseudomonas maltophilia’

    DTIC Science & Technology

    1991-05-31

    Jackson has used plasmid DNA derived from strain 0-2 to transform various metal-resistance phenotypes into a recipient strain of E. coli. He has...PAGE 1 Of AUTRACT’ UNCLASSTJT UNCLASSIFIED UN CLAS SIF IED UL I4N7d.l~0SO Stanbato @Form 29S ( Rev . 24E91 A. RESEARCH OBJECTIVES The aims of this project...Barton and Thomas Zocco at Iew Mexico State University and Los Alamos National Laboratory, respectively. Examination by SEM of cultures of 0-2 grown in the

  4. Transforming growth factor-β: an important mediator in Helicobacter pylori-associated pathogenesis

    PubMed Central

    Li, Nianshuang; Xie, Chuan; Lu, Nong-Hua

    2015-01-01

    Helicobacter pylori (H.pylori) is a Gram-negative, microaerophilic, helical bacillus that specifically colonizes the gastric mucosa. The interaction of virulence factors, host genetic factors, and environmental factors contributes to the pathogenesis of H. pylori-associated conditions, such as atrophic gastritis and intestinal metaplasia. Infection with H. pylori has recently been recognized as the strongest risk factor for gastric cancer. As a pleiotropic cytokine, transforming growth factor (TGF)-β regulates various biological processes, including cell cycle, proliferation, apoptosis, and metastasis. Recent studies have shed new light on the involvement of TGF-β signaling in the pathogenesis of H. pylori infection. This review focuses on the potential etiological roles of TGF-β in H. pylori-mediated gastric pathogenesis. PMID:26583078

  5. Transformation of soybean Gy3 gene into Artemisaarenaria mediated by corona discharge

    NASA Astrophysics Data System (ADS)

    Chao, Lu-meng; Na, Ri; Xue, Dan; Xu, Yongze; Liu, Teng

    2013-03-01

    In order to improve the protein content of desert plant, a method of genetic transformation mediated by corona discharge was established. Artemisia seeds were processed in corona electric field for 120 min at 12 kV, and then soaked in 0.1 SSC media that contained Soybean Gy3 gene DNA to incubate for 12 h at 26 °C. Finally the seeds were inoculated on the differentiation medium. Polymerase Chain Reaction (PCR) and Reverse Transcription Polymerase Chain Reaction (RT-PCR) detection showed that the Soybean Gy3 gene had been successfully introduced into genomic DNA of the regenerated plants of Artemisaarenaria. The study provided a new way for corona discharge in plant genetic modification.

  6. TRANSFORMATION

    SciTech Connect

    LACKS,S.A.

    2003-10-09

    Transformation, which alters the genetic makeup of an individual, is a concept that intrigues the human imagination. In Streptococcus pneumoniae such transformation was first demonstrated. Perhaps our fascination with genetics derived from our ancestors observing their own progeny, with its retention and assortment of parental traits, but such interest must have been accelerated after the dawn of agriculture. It was in pea plants that Gregor Mendel in the late 1800s examined inherited traits and found them to be determined by physical elements, or genes, passed from parents to progeny. In our day, the material basis of these genetic determinants was revealed to be DNA by the lowly bacteria, in particular, the pneumococcus. For this species, transformation by free DNA is a sexual process that enables cells to sport new combinations of genes and traits. Genetic transformation of the type found in S. pneumoniae occurs naturally in many species of bacteria (70), but, initially only a few other transformable species were found, namely, Haemophilus influenzae, Neisseria meningitides, Neisseria gonorrheae, and Bacillus subtilis (96). Natural transformation, which requires a set of genes evolved for the purpose, contrasts with artificial transformation, which is accomplished by shocking cells either electrically, as in electroporation, or by ionic and temperature shifts. Although such artificial treatments can introduce very small amounts of DNA into virtually any type of cell, the amounts introduced by natural transformation are a million-fold greater, and S. pneumoniae can take up as much as 10% of its cellular DNA content (40).

  7. Understanding Dissolved and Colloidal Metal Transport and Transformation - Pathways for Aquatic Toxicity

    NASA Astrophysics Data System (ADS)

    Kimball, B. A.; Besser, J. M.

    2004-05-01

    transformation. Both processes lead to an enrichment of both copper and zinc in the mineral and algal coatings of cobbles and bed sediments where these metals enter the food web through benthic grazing, but the fates of these metals in stream food webs differ substantially. Zinc accumulates to high concentrations in grazing invertebrates but concentrations are substantially lower in higher-order consumers, whereas copper is passed more efficiently to higher order consumers and may pose risks of dietary toxicity to fish. Although this example addresses streams affected by mine drainage, these processes could be active and significant in many other settings where iron or other colloidal transport occurs.

  8. Alcohol-Mediated Resistance-Switching Behavior in Metal-Organic Framework-Based Electronic Devices.

    PubMed

    Liu, Yaqing; Wang, Hong; Shi, Wenxiong; Zhang, Weina; Yu, Jiancan; Chandran, Bevita K; Cui, Chenlong; Zhu, Bowen; Liu, Zhiyuan; Li, Bin; Xu, Cai; Xu, Zhiling; Li, Shuzhou; Huang, Wei; Huo, Fengwei; Chen, Xiaodong

    2016-07-25

    Metal-organic frameworks (MOFs) have drawn increasing attentions as promising candidates for functional devices. Herein, we present MOF films in constructing memory devices with alcohol mediated resistance switching property, where the resistance state is controlled by applying alcohol vapors to achieve multilevel information storage. The ordered packing mode and the hydrogen bonding system of the guest molecules adsorbed in MOF crystals are shown to be the reason for the alcohol mediated electrical switching. This chemically mediated memory device can be a candidate in achieving environment-responsive devices and exhibits potential applications in wearable information storage systems. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. New investigation in PVP-mediated synthesis of noble metallic nanomaterials.

    PubMed

    Zhang, Weijia; Rosano-Ortega, Genoveva; Hu, Ye; Bai, Litao; Qin, Lidong

    2012-03-01

    As one of the most heavily used starting materials in metallic nanostructure syntheses, PVP has made a series of revolutionary successes. However, the true role of PVP still has not been fully understood. Herein, designed reactions and NMR analyses have been done to prove the redox reaction in PVP-mediated synthesis of metallic nanomaterials. As metal ions are reduced, alpha-pyrrolidone rings of PVP are partially oxidized and form poly(vinyl(pyrrolidone)x-(succinimide)y), which has a crucial surface modification capability for nanocrystals. This new finding provides insight into how PVP manipulates the structures and morphologies by modifying the reaction rate and stabilizing the nanocrystals.

  10. Down-regulation of USP13 mediates phenotype transformation of fibroblasts in idiopathic pulmonary fibrosis.

    PubMed

    Geng, Jing; Huang, Xiaoxi; Li, Ying; Xu, Xuefeng; Li, Shuhong; Jiang, Dingyuan; Liang, Jiurong; Jiang, Dianhua; Wang, Chen; Dai, Huaping

    2015-10-09

    Idiopathic pulmonary fibrosis (IPF) is a fatal disease characterized by fibroblastic foci and progressive scarring of the pulmonary parenchyma. IPF fibroblasts display increased proliferation and enhanced migration and invasion, analogous to cancer cells. This transformation-like phenotype of fibroblasts plays an important role in the development of pulmonary fibrosis, but the mechanism for this is not well understood. In this study, we compared gene expression profiles in fibrotic lung tissues from IPF patients and normal lung tissues from patients with primary spontaneous pneumothorax using a cDNA microarray to examine the mechanisms involved in the pathogenesis of IPF. In a cDNA microarray, we found that USP13 was decreased in lung tissues from patients with IPF, which was further confirmed by results from immunohistochemistry and western blot assays. Then, we used RNA interference in MRC-5 cells to inhibit USP13 and evaluated its effects by western blot, real-time RT-PCR, bromodeoxyuridine incorporation, and transwell assays. We also used co-immunoprecipitation and immunofluorescence staining to identify the correlation between USP13 and PTEN in IPF. USP13 expression levels were markedly reduced in fibroblastic foci and primary IPF fibroblast lines. The depletion of USP13 resulted in the transformation of fibroblasts into an aggressive phenotype with enhanced proliferative, migratory, and invasive capacities. Additionally, USP13 interacted with PTEN and mediated PTEN ubiquitination and degradation in lung fibroblasts. Down-regulation of USP13 mediates PTEN protein loss and fibroblast phenotypic change, and thereby plays a crucial role in IPF pathogenesis.

  11. Studies of transformational leadership in consumer service: leadership trust and the mediating-moderating role of cooperative conflict management.

    PubMed

    Yang, Yi-Feng

    2012-02-01

    This is the third in a series of studies evaluating how transformational leadership is associated with related variables such as job satisfaction, change commitment, leadership trust, cooperative conflict management, and market orientation. The present paper evaluates the effects of transformational leadership and cooperative conflict management along with their mediating and moderating of leadership trust in the life insurance industry for two sample groups, sales managers and sales employees. The main effect of leadership trust was mediated and moderated by cooperative conflict management. Cooperative conflict management made a more important contribution than transformational leadership or the moderating effect (interaction), but these three together were the most important variables predicting highest leadership trust. Transformational leadership has an indirect influence on leadership trust. This work summarizes the specific contribution and importance of building successful leadership trust associations with employees in relation to leadership and satisfaction with change commitment.

  12. Agrobacterium-mediated transformation of potato using PLRV-rep and PVY CP genes and assessment of replicase mediated resistance against natural infection of PLRV

    USDA-ARS?s Scientific Manuscript database

    Replicase-and coat protein gene-mediated resistances against potato leafroll virus (PLRV) and potato virus Y (PVY), respectively, demonstrated to be an effective way of protecting potato against two major virus problems (PLRV & PVY) world-wide. Potato cultivar Desiree was transformed using Agrobacte...

  13. Application of sonication in combination with vacuum infiltration enhances the Agrobacterium-mediated genetic transformation in Indian soybean cultivars.

    PubMed

    Arun, Muthukrishnan; Subramanyam, Kondeti; Mariashibu, Thankaraj Salammal; Theboral, Jeevaraj; Shivanandhan, Ganeshan; Manickavasagam, Markandan; Ganapathi, Andy

    2015-02-01

    Soybean is a recalcitrant crop to Agrobacterium-mediated genetic transformation. Development of highly efficient, reproducible, and genotype-independent transformation protocol is highly desirable for soybean genetic improvement. Hence, an improved Agrobacterium-mediated genetic transformation protocol has been developed for cultivar PK 416 by evaluating various parameters including Agrobacterium tumefaciens strains (LBA4404, EHA101, and EHA105 harboring pCAMBIA1304 plasmid), sonication duration, vacuum infiltration pressure, and vacuum duration using cotyledonary node explants of soybean prepared from 7-day-old seedlings. The transformed plants were successfully developed through direct organogenesis system. Transgene expression was assessed by GUS histochemical and gfp visual assays, and integration was analyzed by PCR and Southern blot hybridization. Among the different combinations and durations evaluated, a maximum transformation efficiency of 18.6 % was achieved when the cotyledonary node explants of cv. PK 416 were sonicated for 20 s and vacuum infiltered for 2 min at 250 mmHg in A. tumefaciens EHA105 suspension. The amenability of the standardized protocol was tested on four more soybean cultivars JS 90-41, Hara Soy, Co 1, and Co 2 in which all the cultivars responded favorably with transformation efficiency ranging from 13.3 to 16.6 %. The transformation protocol developed in the present study would be useful to transform diverse soybean cultivars with desirable traits.

  14. Encapsulation of metal clusters within MFI via interzeolite transformations and direct hydrothermal syntheses and catalytic consequences of their confinement.

    PubMed

    Goel, Sarika; Zones, Stacey I; Iglesia, Enrique

    2014-10-29

    The encapsulation of metal clusters (Pt, Ru, Rh) within MFI was achieved by exchanging cationic metal precursors into a parent zeolite (BEA, FAU), reducing them with H2 to form metal clusters, and transforming these zeolites into daughter structures of higher framework density (MFI) under hydrothermal conditions. These transformations required MFI seeds or organic templates for FAU parent zeolites, but not for BEA, and occurred with the retention of encapsulated clusters. Clusters uniform in size (1.3-1.7 nm) and exposing clean and accessible surfaces formed in BEA and FAU zeolites; their size remained essentially unchanged upon transformation into MFI. Encapsulation selectivities, determined from the relative hydrogenation rates of small (toluene) and large (alkyl arenes) molecules and defined as the ratio of the surface areas of all the clusters in the sample to that of external clusters, were very high (8.1-40.9) for both parent and daughter zeolites. Encapsulation into MFI via direct hydrothermal syntheses was unsuccessful because metal precursors precipitated prematurely at the pH and temperatures required for MFI synthesis. Delayed introduction of metal precursors and F(-) (instead of OH(-)) as the mineralizing agent in hydrothermal syntheses increased encapsulation selectivities, but they remained lower than those achieved via interzeolite transformations. These interconversions provide a general and robust strategy for encapsulation of metals when precursors can be introduced via exchange into a zeolite that can be transformed into target daughter zeolites with higher framework densities, whether spontaneously or by using seeds or structure-directing agents (SDA).

  15. Contiguous metal-mediated base pairs comprising two Ag(I) ions.

    PubMed

    Megger, Dominik A; Guerra, Célia Fonseca; Hoffmann, Jan; Brutschy, Bernhard; Bickelhaupt, F Matthias; Müller, Jens

    2011-05-27

    The incorporation of transition-metal ions into nucleic acids by using metal-mediated base pairs has proved to be a promising strategy for the site-specific functionalization of these biomolecules. We report herein the formation of Ag(+)-mediated Hoogsteen-type base pairs comprising 1,3-dideaza-2'-deoxyadenosine and thymidine. By defunctionalizing the Watson-Crick edge of adenine, the formation of regular base pairs is prohibited. The additional substitution of the N3 nitrogen atom of adenine by a methine moiety increases the basicity of the exocyclic amino group. Hence, 1,3-dideazaadenine and thymine are able to incorporate two Ag(+) ions into their Hoogsteen-type base pair (as compared with one Ag(+) ion in base pairs with 1-deazaadenine and thymine). We show by using a combination of experimental techniques (UV and circular dichroism (CD) spectroscopies, dynamic light scattering, and mass spectrometry) that this type of base pair is compatible with different sequence contexts and can be used contiguously in DNA double helices. The most stable duplexes were observed when using a sequence containing alternating purine and pyrimidine nucleosides. Dispersion-corrected density functional theory calculations have been performed to provide insight into the structure, formation and stabilization of the twofold metalated base pair. They revealed that the metal ions within a base pair are separated by an Ag···Ag distance of about 2.88 Å. The Ag-Ag interaction contributes some 16 kcal mol(-1) to the overall stability of the doubly metal-mediated base pair, with the dominant contribution to the Ag-Ag bonding resulting from a donor-acceptor interaction between silver 4d-type and 4s orbitals. These Hoogsteen-type base pairs enable a higher functionalization of nucleic acids with metal ions than previously reported metal-mediated base pairs, thereby increasing the potential of DNA-based nanotechnology.

  16. Fusion tyrosine kinase mediated signalling pathways in the transformation of haematopoietic cells.

    PubMed

    Turner, S D; Alexander, D R

    2006-04-01

    The fusion tyrosine kinases (FTKs) are generated by chromosomal translocations creating bipartite proteins in which the kinase is hyperactivated by an adjoining oligomerization domain. Autophosphorylation of the FTK generates a 'signalosome', an ensemble of signalling proteins that transduce signals to downstream pathways. At the earliest stages of oncogenesis, FTKs can mimic mitogenic cytokine signalling pathways involving the GAB-2 adaptor protein and signal transducers and activators of transcription (STAT) factors, generating replicative stress and thereby promoting a mutator phenotype. In parallel, FTKs couple to survival pathways that upregulate prosurvival proteins such as Bcl-xL, so preventing DNA-damage-induced apoptosis. Following transformation, FTKs induce resistance to genotoxic attack by upregulating DNA repair mechanisms such as STAT5-dependent RAD51 transcription. The phenomenon of 'oncogene addiction' reflects the continued requirement of an active FTK 'signalosome' to mediate survival and mitogenic signals involving the PI 3-kinase and mitogen-activated protein stress-activated protein kinase pathways, and the nuclear factor-kappa B, activator protein 1 and STAT transcription factors. The available data so far suggest that FTKs, with some possible exceptions, induce and maintain the transformed state using similar panoplies of signals, a finding with important therapeutic implications. The FTK signalling field has matured to an exciting phase in which rapid advances are facilitating rational drug design.

  17. Mediation of wound-related Rous sarcoma virus tumorigenesis by TFG (transforming growth factor)-. beta

    SciTech Connect

    Sieweke, M.H.; Bissell, M.J. ); Thompson, N.L.; Sporn, M.B. )

    1990-06-29

    In Rous sarcoma virus (RSV)-infected chickens, wounding leads to tumor formation with nearly 100% frequency in tissues that would otherwise remain tumor-free. Identifying molecular mediators of this phenomenon should yield important clues to the mechanisms involved in RSV tumorigenesis. Immunohistochemical staining showed that TGF-{beta} is present locally shortly after wounding, but not in unwounded controls. In addition, subcutaneous administration of recombinant transforming growth factor {beta}1 (TGF-{beta}1) could substitute completely for wounding in tumor induction. A treatment protocol of four doses of 800 nanograms of TGF-{beta} resulted in v-src-expressing tumors with 100% frequency; four doses of only 10 nanograms still led to tumor formation in 80% of the animals. This effect was specific, as other growth factors with suggested roles in would healing did not elicit the same response. Epidermal growth factor (EGF) or TGF-{alpha} had no effect, and platelet-derived growth factor (PDGF) or insulin-like growth factor-1 (IGF-1) yielded only occasional tumors after longer latency. TGF-{beta} release during the would-healing response may thus be a critical event that creates a conducive environment for RSV tumorigenesis and may act as a cofactor for transformation in this system. 31 refs., 3 figs., 2 tabs.

  18. Transformed Drosophila Cells Evade Diet-Mediated Insulin Resistance Through Wingless Signaling

    PubMed Central

    Hirabayashi, Susumu; Baranski, Thomas J.; Cagan, Ross L.

    2013-01-01

    SUMMARY Risk of specific cancers increases in patients with metabolic dysfunction including obesity and diabetes. Here we use Drosophila as a model to explore the effects of diet on tumor progression. Feeding Drosophila a diet high in carbohydrates was previously demonstrated to direct metabolic dysfunction including hyperglycemia, hyperinsulinemia and insulin-resistance. We demonstrate that high dietary sugar also converts Ras/Src transformed tissue from localized growths to aggressive tumors with emergent metastases. While most tissues displayed insulin resistance, Ras/Src tumors retained insulin pathway sensitivity, increased the ability to import glucose, and resisted apoptosis. High dietary sugar increased canonical Wingless/Wnt pathway activity, which upregulated Insulin Receptor gene expression to promote insulin sensitivity. The result is a feed-forward circuit that amplified diet-mediated malignant phenotypes within Ras/Src transformed tumors. By targeting multiple steps in this circuit with rationally applied drug combinations, we demonstrate the potential of combinatorial drug intervention to treat diet-enhanced malignant tumors. PMID:23911328

  19. Spectral induced polarization and electrodic potential monitoring of microbially mediated iron sulfide transformations

    SciTech Connect

    Hubbard, Susan; Personna, Y.R.; Ntarlagiannis, D.; Slater, L.; Yee, N.; O'Brien, M.; Hubbard, S.

    2008-02-15

    Stimulated sulfate-reduction is a bioremediation technique utilized for the sequestration of heavy metals in the subsurface.We performed laboratory column experiments to investigate the geoelectrical response of iron sulfide transformations by Desulfo vibriovulgaris. Two geoelectrical methods, (1) spectral induced polarization (SIP), and (2) electrodic potential measurements, were investigated. Aqueous geochemistry (sulfate, lactate, sulfide, and acetate), observations of precipitates (identified from electron microscopy as iron sulfide), and electrodic potentials on bisulfide ion (HS) sensitive silver-silver chloride (Ag-AgCl) electrodes (630 mV) were diagnostic of induced transitions between an aerobic iron sulfide forming conditions and aerobic conditions promoting iron sulfide dissolution. The SIP data showed 10m rad anomalies during iron sulfide mineralization accompanying microbial activity under an anaerobic transition. These anomalies disappeared during iron sulfide dissolution under the subsequent aerobic transition. SIP model parameters based on a Cole-Cole relaxation model of the polarization at the mineral-fluid interface were converted to (1) estimated biomineral surface area to pore volume (Sp), and (2) an equivalent polarizable sphere diameter (d) controlling the relaxation time. The temporal variation in these model parameters is consistent with filling and emptying of pores by iron sulfide biofilms, as the system transitions between anaerobic (pore filling) and aerobic (pore emptying) conditions. The results suggest that combined SIP and electrodic potential measurements might be used to monitor spatiotemporal variability in microbial iron sulfide transformations in the field.

  20. Spectral induced polarization and electrodic potential monitoring of microbially mediated iron sulfide transformations

    NASA Astrophysics Data System (ADS)

    Personna, Yves Robert; Ntarlagiannis, Dimitrios; Slater, Lee; Yee, Nathan; O'Brien, Michael; Hubbard, Susan

    2008-06-01

    Stimulated sulfate-reduction is a bioremediation technique utilized for the sequestration of heavy metals in the subsurface. We performed laboratory column experiments to investigate the geoelectrical response of iron sulfide transformations by Desulfovibrio vulgaris. Two geoelectrical methods, (1) spectral induced polarization (SIP), and (2) electrodic potential measurements, were investigated. Aqueous geochemistry (sulfate, lactate, sulfide, and acetate), observations of precipitates (identified from electron microscopy as iron sulfide), and electrodic potentials on bisulfide ion (HS-) sensitive silver-silver chloride (Ag-AgCl) electrodes (˜-630 mV) were diagnostic of induced transitions between anaerobic iron sulfide forming conditions and aerobic conditions promoting iron sulfide dissolution. The SIP data showed ˜10 mrad anomalies during iron sulfide mineralization accompanying microbial activity under an anaerobic transition. These anomalies disappeared during iron sulfide dissolution under the subsequent aerobic transition. SIP model parameters based on a Cole-Cole relaxation model of the polarization at the mineral-fluid interface were converted to (1) estimated biomineral surface area to pore volume (Sp), and (2) an equivalent polarizable sphere diameter (d) controlling the relaxation time. The temporal variation in these model parameters is consistent with filling and emptying of pores by iron sulfide biofilms, as the system transitions between anaerobic (pore filling) and aerobic (pore emptying) conditions. The results suggest that combined SIP and electrodic potential measurements might be used to monitor spatiotemporal variability in microbial iron sulfide transformations in the field.

  1. Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L. x P. maximowiczii A. Henry.

    PubMed

    Yevtushenko, Dmytro P; Misra, Santosh

    2010-03-01

    Many economically important species of Populus, especially those in sections Aigeiros and Tacamahaca, remain recalcitrant to genetic transformation. In this study, a simple and reliable protocol was developed for the efficient Agrobacterium-mediated transformation of a difficult-to-transform, but commercially viable, hybrid poplar Populus nigra L. x P. maximowiczii A. Henry (NM6). A plant transformation vector designed to express the beta-glucuronidase (GUS) gene was used to detect transformation events at early stages of plant regeneration and to optimize parameters affecting poplar transformation. The use of zeatin riboside in shoot-induction medium, regeneration of shoots via indirect organogenesis, and early selection pressure were the major modifications that drastically improved the efficiency of poplar transformation and minimized the number of untransformed regenerants. Transgenic shoots were routinely obtained 4-10 weeks after co-culture with A. tumefaciens, with a greater than 90% rate of plant recovery. Stable transgene integration, ranging from a single insertion to ten copies per genome, was confirmed by Southern blot analysis. The mean transformation frequency was 36.3% and about two-thirds of the lines had 1-2 transgene copies. Among the explants, petioles and leaves had a higher transformation frequency than did stem segments. Growth characteristics and the morphology of transgenic poplar plants were identical to untransformed controls. These findings will accelerate the development of P. nigra x P. maximowiczii plants with novel traits, and may also be useful to improve transformation procedures for other Populus species.

  2. X-ray induced, substrate-carrier mediated deposition of metal on GaAs

    SciTech Connect

    Ma Qing; Divan, R.; Mancini, D. C.; Rosenberg, R. A.; Quintana, J. P.; Keane, D. T.

    2006-08-21

    A wet metal deposition process on GaAs surfaces is described. The process is induced by high energy x-ray photons and is mediated by photon-generated carriers through the photoelectrochemical mechanism similar to that for light-induced wet etching. The micrometer to submicrometer feature fabrication using this process is demonstrated.

  3. Peroxide-mediated transition-metal-free direct amidation of alcohols with nitroarenes.

    PubMed

    Xiao, Fuhong; Liu, Yong; Tang, Chenglin; Deng, Guo-Jun

    2012-02-17

    An unusual direct amidation of alcohols with nitroarenes mediated by peroxides has been discovered. The reaction tolerated a wide range of functionalities, and various aromatic amides were obtained in moderate to good yields in the absence of transition-metal catalyst. The peroxides and solvents had a significant impact on the reaction yield.

  4. TRANSFORMER

    DOEpatents

    Baker, W.R.

    1959-08-25

    Transformers of a type adapted for use with extreme high power vacuum tubes where current requirements may be of the order of 2,000 to 200,000 amperes are described. The transformer casing has the form of a re-entrant section being extended through an opening in one end of the cylinder to form a coaxial terminal arrangement. A toroidal multi-turn primary winding is disposed within the casing in coaxial relationship therein. In a second embodiment, means are provided for forming the casing as a multi-turn secondary. The transformer is characterized by minimized resistance heating, minimized external magnetic flux, and an economical construction.

  5. Agrobacterium-mediated genetic transformation of yam (Dioscorea rotundata): an important tool for functional study of genes and crop improvement

    PubMed Central

    Nyaboga, Evans; Tripathi, Jaindra N.; Manoharan, Rajesh; Tripathi, Leena

    2014-01-01

    Although genetic transformation of clonally propagated crops has been widely studied as a tool for crop improvement and as a vital part of the development of functional genomics resources, there has been no report of any existing Agrobacterium-mediated transformation of yam (Dioscorea spp.) with evidence of stable integration of T-DNA. Yam is an important crop in the tropics and subtropics providing food security and income to over 300 million people. However, yam production remains constrained by increasing levels of field and storage pests and diseases. A major constraint to the development of biotechnological approaches for yam improvement has been the lack of an efficient and robust transformation and regeneration system. In this study, we developed an Agrobacterium-mediated transformation of Dioscorea rotundata using axillary buds as explants. Two cultivars of D. rotundata were transformed using Agrobacterium tumefaciens harboring the binary vectors containing selectable marker and reporter genes. After selection with appropriate concentrations of antibiotic, shoots were developed on shoot induction and elongation medium. The elongated antibiotic-resistant shoots were subsequently rooted on medium supplemented with selection agent. Successful transformation was confirmed by polymerase chain reaction, Southern blot analysis, and reporter genes assay. Expression of gusA gene in transgenic plants was also verified by reverse transcription polymerase chain reaction analysis. Transformation efficiency varied from 9.4 to 18.2% depending on the cultivars, selectable marker genes, and the Agrobacterium strain used for transformation. It took 3–4 months from Agro-infection to regeneration of complete transgenic plant. Here we report an efficient, fast and reproducible protocol for Agrobacterium-mediated transformation of D. rotundata using axillary buds as explants, which provides a useful platform for future genetic engineering studies in this economically important

  6. Controllable Phase Transformation in Extracting Valuable Metals from Chinese Low-Grade Nickel Sulphide Ore

    NASA Astrophysics Data System (ADS)

    Cui, Fuhui; Mu, Wenning; Wang, Shuai; Xu, Qian; Zhai, Yuchun; Luo, Shaohua

    2017-10-01

    In this work, a two-stage sulphuric acid roasting and water leaching system was chosen to extract valuable metals from Chinese low-grade nickel sulphide ore. By optimizing the two-stage roasting process, first roasting temperature at 295°C with particle size of 80-91 μm with an acid-ore ratio of 1.1:1 for 2 h, and second roasting temperature at 620°C for 2 h, it was found that more than 98% of the nickel and 99% of the copper but less than 14.38% of Fe were leached into the water. Attempts were made via x-ray diffraction analysis, scanning electron microscopy, chemical phase analysis, and differential thermal and thermogravimetric analysis to reveal the phase transformations for Ni, Cu, and Mg, which could be expressed as mineral phases → sulphates hydrate → sulphates and for iron as mineral phases → hydrated ferrous sulphate → ferric sulphates and ferric oxide → oxide. The results of this work suggest that a controllable phase transformation by using a two-stage sulphuric acid roasting process is a feasible method for efficiently extracting valuable metals from Chinese nickel sulphide ore.

  7. Abnormal correlation between phase transformation and cooling rate for pure metals

    PubMed Central

    Han, J. J.; Wang, C. P.; Liu, X. J.; Wang, Y.; Liu, Z.-K.; Zhang, T.-Y.; Jiang, J. Z.

    2016-01-01

    This work aims to achieve deep insight into the phenomenon of phase transformation upon rapid cooling in metal systems and reveal the physical meaning of scatter in the time taken to reach crystallization. The total number of pure metals considered in this work accounts for 14. Taking pure copper as an example, the correlation between phase selection of crystal or glass and cooling rate was investigated using molecular dynamic simulations. The obtained results demonstrate that there exists a cooling rate region of 6.3 × 1011–16.6 × 1011 K/s, in which crystalline fractions largely fluctuate along with cooling rates. Glass transformation in this cooling rate region is determined by atomic structure fluctuation, which is controlled by thermodynamic factors. According to the feature of bond-orientation order at different cooling rates, we propose two mechanisms of glass formation: (i) kinetic retardation of atom rearrangement or structural relaxation at a high cooling rate; and (ii) competition of icosahedral order against crystal order near the critical cooling rate. PMID:26939584

  8. Abnormal correlation between phase transformation and cooling rate for pure metals.

    PubMed

    Han, J J; Wang, C P; Liu, X J; Wang, Y; Liu, Z-K; Zhang, T-Y; Jiang, J Z

    2016-03-04

    This work aims to achieve deep insight into the phenomenon of phase transformation upon rapid cooling in metal systems and reveal the physical meaning of scatter in the time taken to reach crystallization. The total number of pure metals considered in this work accounts for 14. Taking pure copper as an example, the correlation between phase selection of crystal or glass and cooling rate was investigated using molecular dynamic simulations. The obtained results demonstrate that there exists a cooling rate region of 6.3 × 10(11)-16.6 × 10(11) K/s, in which crystalline fractions largely fluctuate along with cooling rates. Glass transformation in this cooling rate region is determined by atomic structure fluctuation, which is controlled by thermodynamic factors. According to the feature of bond-orientation order at different cooling rates, we propose two mechanisms of glass formation: (i) kinetic retardation of atom rearrangement or structural relaxation at a high cooling rate; and (ii) competition of icosahedral order against crystal order near the critical cooling rate.

  9. Metal-induced rapid transformation of diamond into single and multilayer graphene on wafer scale

    DOE PAGES

    Berman, Diana; Deshmukh, Sanket; Narayanan, Badri; ...

    2016-07-04

    The degradation of intrinsic properties of graphene during the transfer process constitutes a major challenge in graphene device fabrication, stimulating the need for direct growth of graphene on dielectric substrates. Previous attempts of metal-induced transformation of diamond and silicon carbide into graphene suffers from metal contamination and inability to scale graphene growth over large area. Here in this article, we introduce a direct approach to transform polycrystalline diamond into high-quality graphene layers on wafer scale (4 inch in diameter) using a rapid thermal annealing process facilitated by a nickel, Ni thin film catalyst on top. We show that the processmore » can be tuned to grow single or multilayer graphene with good electronic properties. Molecular dynamics simulations elucidate the mechanism of graphene growth on polycrystalline diamond. Additionally, we demonstrate the lateral growth of free-standing graphene over micron-sized pre-fabricated holes, opening exciting opportunities for future graphene/diamond-based electronics.« less

  10. Controllable Phase Transformation in Extracting Valuable Metals from Chinese Low-Grade Nickel Sulphide Ore

    NASA Astrophysics Data System (ADS)

    Cui, Fuhui; Mu, Wenning; Wang, Shuai; Xu, Qian; Zhai, Yuchun; Luo, Shaohua

    2017-06-01

    In this work, a two-stage sulphuric acid roasting and water leaching system was chosen to extract valuable metals from Chinese low-grade nickel sulphide ore. By optimizing the two-stage roasting process, first roasting temperature at 295°C with particle size of 80-91 μm with an acid-ore ratio of 1.1:1 for 2 h, and second roasting temperature at 620°C for 2 h, it was found that more than 98% of the nickel and 99% of the copper but less than 14.38% of Fe were leached into the water. Attempts were made via x-ray diffraction analysis, scanning electron microscopy, chemical phase analysis, and differential thermal and thermogravimetric analysis to reveal the phase transformations for Ni, Cu, and Mg, which could be expressed as mineral phases → sulphates hydrate → sulphates and for iron as mineral phases → hydrated ferrous sulphate → ferric sulphates and ferric oxide → oxide. The results of this work suggest that a controllable phase transformation by using a two-stage sulphuric acid roasting process is a feasible method for efficiently extracting valuable metals from Chinese nickel sulphide ore.

  11. Homogeneous and Heterogeneous Reaction and Transformation of Hg and Trace Metals in Combustion Systems

    SciTech Connect

    J. Helble; Clara Smith; David Miller

    2009-08-31

    The overall goal of this project was to produce a working dynamic model to predict the transformation and partitioning of trace metals resulting from combustion of a broad range of fuels. The information provided from this model will be instrumental in efforts to identify fuels and conditions that can be varied to reduce metal emissions. Through the course of this project, it was determined that mercury (Hg) and arsenic (As) would be the focus of the experimental investigation. Experiments were therefore conducted to examine homogeneous and heterogeneous mercury oxidation pathways, and to assess potential interactions between arsenic and calcium. As described in this report, results indicated that the role of SO{sub 2} on Hg oxidation was complex and depended upon overall gas phase chemistry, that iron oxide (hematite) particles contributed directly to heterogeneous Hg oxidation, and that As-Ca interactions occurred through both gas-solid and within-char reaction pathways. Modeling based on this study indicated that, depending upon coal type and fly ash particle size, vaporization-condensation, vaporization-surface reaction, and As-CaO in-char reaction all play a role in arsenic transformations under combustion conditions.

  12. Alkali metal mediated C–C bond coupling reaction

    SciTech Connect

    Tachikawa, Hiroto

    2015-02-14

    Metal catalyzed carbon-carbon (C–C) bond formation is one of the important reactions in pharmacy and in organic chemistry. In the present study, the electron and hole capture dynamics of a lithium-benzene sandwich complex, expressed by Li(Bz){sub 2}, have been investigated by means of direct ab-initio molecular dynamics method. Following the electron capture of Li(Bz){sub 2}, the structure of [Li(Bz){sub 2}]{sup −} was drastically changed: Bz–Bz parallel form was rapidly fluctuated as a function of time, and a new C–C single bond was formed in the C{sub 1}–C{sub 1}′ position of Bz–Bz interaction system. In the hole capture, the intermolecular vibration between Bz–Bz rings was only enhanced. The mechanism of C–C bond formation in the electron capture was discussed on the basis of theoretical results.

  13. Surface-mediated light transmission in metal nanoparticle chains

    NASA Astrophysics Data System (ADS)

    Compaijen, P. Jasper; Malyshev, Victor A.; Knoester, Jasper

    2013-05-01

    We study theoretically the efficiency of the transmission of optical signals through a linear chain consisting of identical and equidistantly spaced silver metal nanoparticles. Two situations are compared: the transmission efficiency through an isolated chain and through a chain in close proximity of a reflecting substrate. The Ohmic and radiative losses in each nanoparticle strongly affect the transmission efficiency of an isolated chain and suppress it to large extent. It is shown that the presence of a reflecting interface may enhance the guiding properties of the array. The reason for this is the energy exchange between the surface plasmon polaritons (SPPs) of the array and the substrate. We focus on the dependence of the transmission efficiency on the frequency and polarization of the incoming light, as well as on the influence of the array-interface spacing. Sometimes the effect of these parameters turns out to be counterintuitive, reflecting a complicated interplay of several transmission channels.

  14. Alkali metal mediated C-C bond coupling reaction

    NASA Astrophysics Data System (ADS)

    Tachikawa, Hiroto

    2015-02-01

    Metal catalyzed carbon-carbon (C-C) bond formation is one of the important reactions in pharmacy and in organic chemistry. In the present study, the electron and hole capture dynamics of a lithium-benzene sandwich complex, expressed by Li(Bz)2, have been investigated by means of direct ab-initio molecular dynamics method. Following the electron capture of Li(Bz)2, the structure of [Li(Bz)2]- was drastically changed: Bz-Bz parallel form was rapidly fluctuated as a function of time, and a new C-C single bond was formed in the C1-C1' position of Bz-Bz interaction system. In the hole capture, the intermolecular vibration between Bz-Bz rings was only enhanced. The mechanism of C-C bond formation in the electron capture was discussed on the basis of theoretical results.

  15. Alkali metal mediated C-C bond coupling reaction.

    PubMed

    Tachikawa, Hiroto

    2015-02-14

    Metal catalyzed carbon-carbon (C-C) bond formation is one of the important reactions in pharmacy and in organic chemistry. In the present study, the electron and hole capture dynamics of a lithium-benzene sandwich complex, expressed by Li(Bz)2, have been investigated by means of direct ab-initio molecular dynamics method. Following the electron capture of Li(Bz)2, the structure of [Li(Bz)2](-) was drastically changed: Bz-Bz parallel form was rapidly fluctuated as a function of time, and a new C-C single bond was formed in the C1-C1' position of Bz-Bz interaction system. In the hole capture, the intermolecular vibration between Bz-Bz rings was only enhanced. The mechanism of C-C bond formation in the electron capture was discussed on the basis of theoretical results.

  16. [Influence of sulfur on the speciation transformation and phyto-availability of heavy metals in soil: a review].

    PubMed

    Sun, Li-Juan; Duan, De-Chao; Peng, Cheng; He, Jun-Yu; Shi, Ji-Yan

    2014-07-01

    The biogeochemical cycle of sulfur can directly affect the speciation transformation of heavy metals in soils and their accumulation in plants. The toxicity of heavy metals in plants can be alleviated by their complexation with sulfur compounds like phytochelatins or metallothiones, consisting of the major mechanisms of phytodetoxification. Sulfur deficiency is becoming one of the limiting factors that influence Chinese agricultural production. Although the applications of sulfur fertilizer in soil have received much attention in recent years, the interaction mechanism of heavy metal with sulfur metabolism has not been studied extensively. In this paper, we reviewed current research advance on the impact of sulfur on the speciation transformation of soil heavy metals and their accumulation in plants, discussed the effect of sulfur on the detoxification mechanism of heavy metal in plants and provided further research prospective in this field.

  17. Metal-mediated aminocatalysis provides mild conditions: Enantioselective Michael addition mediated by primary amino catalysts and alkali-metal ions.

    PubMed

    Leven, Matthias; Neudörfl, Jörg M; Goldfuss, Bernd

    2013-01-01

    Four catalysts based on new amides of chiral 1,2-diamines and 2-sulfobenzoic acid have been developed. The alkali-metal salts of these betaine-like amides are able to form imines with enones, which are activated by Lewis acid interaction for nucleophilic attack by 4-hydroxycoumarin. The addition of 4-hydroxycoumarin to enones gives ee's up to 83% and almost quantitative yields in many cases. This novel type of catalysis provides an effective alternative to conventional primary amino catalysis were strong acid additives are essential components.

  18. Metal-mediated aminocatalysis provides mild conditions: Enantioselective Michael addition mediated by primary amino catalysts and alkali-metal ions

    PubMed Central

    Leven, Matthias; Neudörfl, Jörg M

    2013-01-01

    Summary Four catalysts based on new amides of chiral 1,2-diamines and 2-sulfobenzoic acid have been developed. The alkali-metal salts of these betaine-like amides are able to form imines with enones, which are activated by Lewis acid interaction for nucleophilic attack by 4-hydroxycoumarin. The addition of 4-hydroxycoumarin to enones gives ee’s up to 83% and almost quantitative yields in many cases. This novel type of catalysis provides an effective alternative to conventional primary amino catalysis were strong acid additives are essential components. PMID:23400419

  19. Lower nanometer-scale size limit for the deformation of a metallic glass by shear transformations revealed by quantitative AFM indentation

    PubMed Central

    Bennewitz, Roland

    2015-01-01

    Summary We combine non-contact atomic force microscopy (AFM) imaging and AFM indentation in ultra-high vacuum to quantitatively and reproducibly determine the hardness and deformation mechanisms of Pt(111) and a Pt57.5Cu14.7Ni5.3P22.5 metallic glass with unprecedented spatial resolution. Our results on plastic deformation mechanisms of crystalline Pt(111) are consistent with the discrete mechanisms established for larger scales: Plasticity is mediated by dislocation gliding and no rate dependence is observed. For the metallic glass we have discovered that plastic deformation at the nanometer scale is not discrete but continuous and localized around the indenter, and does not exhibit rate dependence. This contrasts with the observation of serrated, rate-dependent flow of metallic glasses at larger scales. Our results reveal a lower size limit for metallic glasses below which shear transformation mechanisms are not activated by indentation. In the case of metallic glass, we conclude that the energy stored in the stressed volume during nanometer-scale indentation is insufficient to account for the interfacial energy of a shear band in the glassy matrix. PMID:26425424

  20. Lower nanometer-scale size limit for the deformation of a metallic glass by shear transformations revealed by quantitative AFM indentation.

    PubMed

    Caron, Arnaud; Bennewitz, Roland

    2015-01-01

    We combine non-contact atomic force microscopy (AFM) imaging and AFM indentation in ultra-high vacuum to quantitatively and reproducibly determine the hardness and deformation mechanisms of Pt(111) and a Pt57.5Cu14.7Ni5.3P22.5 metallic glass with unprecedented spatial resolution. Our results on plastic deformation mechanisms of crystalline Pt(111) are consistent with the discrete mechanisms established for larger scales: Plasticity is mediated by dislocation gliding and no rate dependence is observed. For the metallic glass we have discovered that plastic deformation at the nanometer scale is not discrete but continuous and localized around the indenter, and does not exhibit rate dependence. This contrasts with the observation of serrated, rate-dependent flow of metallic glasses at larger scales. Our results reveal a lower size limit for metallic glasses below which shear transformation mechanisms are not activated by indentation. In the case of metallic glass, we conclude that the energy stored in the stressed volume during nanometer-scale indentation is insufficient to account for the interfacial energy of a shear band in the glassy matrix.

  1. Agrobacterium-mediated high-frequency transformation of an elite commercial maize (Zea mays L.) inbred line.

    PubMed

    Cho, Myeong-Je; Wu, Emily; Kwan, Jackie; Yu, Maryanne; Banh, Jenny; Linn, Wutt; Anand, Ajith; Li, Zhi; TeRonde, Susan; Register, James C; Jones, Todd J; Zhao, Zuo-Yu

    2014-10-01

    An improved Agrobacterium -mediated transformation protocol is described for a recalcitrant commercial maize elite inbred with optimized media modifications and AGL1. These improvements can be applied to other commercial inbreds. This study describes a significantly improved Agrobacterium-mediated transformation protocol in a recalcitrant commercial maize elite inbred, PHR03, using optimal co-cultivation, resting and selection media. The use of green regenerative tissue medium components, high copper and 6-benzylaminopurine, in resting and selection media dramatically increased the transformation frequency. The use of glucose in resting medium further increased transformation frequency by improving the tissue induction rate, tissue survival and tissue proliferation from immature embryos. Consequently, an optimal combination of glucose, copper and cytokinin in the co-cultivation, resting and selection media resulted in significant improvement from 2.6 % up to tenfold at the T0 plant level using Agrobacterium strain LBA4404 in transformation of PHR03. Furthermore, we evaluated four different Agrobacterium strains, LBA4404, AGL1, EHA105, and GV3101 for transformation frequency and event quality. AGL1 had the highest transformation frequency with up to 57.1 % at the T0 plant level. However, AGL1 resulted in lower quality events (defined as single copy for transgenes without Agrobacterium T-DNA backbone) when compared to LBA4404 (30.1 vs 25.6 %). We propose that these improvements can be applied to other recalcitrant commercial maize inbreds.

  2. Linking transformational leadership to nurses' extra-role performance: the mediating role of self-efficacy and work engagement.

    PubMed

    Salanova, Marisa; Lorente, Laura; Chambel, Maria J; Martínez, Isabel M

    2011-10-01

    This paper is a report of a social cognitive theory-guided study about the link between supervisors' transformational leadership and staff nurses' extra-role performance as mediated by nurse self-efficacy and work engagement. Past research has acknowledged the positive influence that transformational leaders have on employee (extra-role) performance. However, less is known about the psychological mechanisms that may explain the links between transformational leaders and extra-role performance, which encompasses behaviours that are not considered formal job requirements, but which facilitate the smooth functioning of the organization as a social system. Seventeen supervisors evaluated nurses' extra-role performance, the data generating a sample consisting of 280 dyads. The nurses worked in different health services in a large Portuguese hospital and the participation rate was 76·9% for nurses and 100% for supervisors. Data were collected during 2009. A theory-driven model of the relationships between transformation leadership, self-efficacy, work engagement and nurses' extra-role performance was tested using Structural Equation Modelling. Data analysis revealed a full mediation model in which transformational leadership explained extra-role performance through self-efficacy and work engagement. A direct relationship between transformational leadership and work engagement was also found. Nurses' supervisors with a transformational leadership style enhance different 'extra-role' performance in nurses and this increases hospital efficacy. They do so by establishing a sense of self-efficacy but also by amplifying their levels of engagement in the workplace. © 2011 Blackwell Publishing Ltd.

  3. Agrobacterium tumefaciens-mediated transformation as an efficient tool for insertional mutagenesis of Cercospora zeae-maydis.

    PubMed

    Lu, Yuanyuan; Xiao, Shuqin; Wang, Fen; Sun, Jiaying; Zhao, Likun; Yan, Libin; Xue, Chunsheng

    2017-02-01

    An efficient Agrobacterium tumefaciens-mediated transformation (ATMT) approach was developed for the plant pathogenic fungus, Cercospora zeae-maydis, which is the causative agent of gray leaf spot in maize. The transformation was evaluated with five parameters to test the efficiencies of transformation. Results showed that spore germination time, co-cultivation temperature and time were the significant influencing factors in all parameters. Randomly selected transformants were confirmed and the transformants were found to be mitotically stable, with single-copy T-DNA integration in the genome. T-DNA flanking sequences were cloned by thermal asymmetric interlaced PCR. Thus, the ATMT approach is an efficient tool for insertional mutagenesis of C. zeae-maydis.

  4. Bioavailable transition metals in particulate matter mediate cardiopulmonary injury in healthy and compromised animal models.

    PubMed Central

    Costa, D L; Dreher, K L

    1997-01-01

    Many epidemiologic reports associate ambient levels of particulate matter (PM) with human mortality and morbidity, particularly in people with preexisting cardiopulmonary disease (e.g., chronic obstructive pulmonary disease, infection, asthma). Because much ambient PM is derived from combustion sources, we tested the hypothesis that the health effects of PM arise from anthropogenic PM that contains bioavailable transition metals. The PM samples studied derived from three emission sources (two oil and one coal fly ash) and four ambient airsheds (St. Louis, MO; Washington; Dusseldorf, Germany; and Ottawa, Canada). PM was administered to rats by intratracheal instillation in equimass or equimetal doses to address directly the influence of PM mass versus metal content on acute lung injury and inflammation. Our results indicated that the lung dose of bioavailable transition metal, not instilled PM mass, was the primary determinant of the acute inflammatory response for both the combustion source and ambient PM samples. Residual oil fly ash, a combustion PM rich in bioavailable metal, was evaluated in a rat model of cardiopulmonary disease (pulmonary vasculitis/hypertension) to ascertain whether the disease state augmented sensitivity to that PM. Significant mortality and enhanced airway responsiveness were observed. Analysis of the lavaged lung fluids suggested that the milieu of the inflamed lung amplified metal-mediated oxidant chemistry to jeopardize the compromised cardiopulmonary system. We propose that soluble metals from PM mediate the array of PM-associated injuries to the cardiopulmonary system of the healthy and at-risk compromised host. PMID:9400700

  5. Metal-mediated molecular materials at the nano- and mesoscale

    NASA Astrophysics Data System (ADS)

    Arroyo, Itzia Zoraida

    The synthesis of materials via self-assembly is a powerful bottom-up approach for assembling matter from subnanometer up to micrometer scales. This methodology involves the spontaneous and reversible organization of small molecules to create larger structures driven by non-covalent interactions such as hydrogen bonding, hydrophobic forces and metal-ligand coordination interactions. In this dissertation we developed the synthetic methods to generate materials at the nano- and meso-scale using coordination-directed strategies for molecular self-assembly in solid-state and in water. In addition, we produced materials with a modular increased complexity with potential applications in advanced technologies and medicine. Molecular materials in the solid-state were engineered using the coordination directed approach by synthesizing organic ligands with well-defined geometries and symmetries that self-assembly with transition metals in aprotic media into supra-molecular arrays. These structures were crystallized and characterized by techniques such as X-ray Crystallography, Multi-Nuclear Magnetic Resonance (NMR), Mass Spectrometry (MS), Infrared (IR) and Ultraviolet-Visible (UV-vis) Spectroscopies. Potential application as hydrogen storage systems was evaluated using 2H NMR spectroscopy. Coordination-directed molecular materials that self-assembly in water were achieved by combining coordination capable amphiphilic molecules and designing their chemistry so that they can rearrange in water to produce different lyotropic phases. We characterized these materials using Extended X-ray Absorbance Fine Structure Spectroscopy (EXAFS), Dynamic Light Scattering, Atomic Force Microscopy (AFM), Transmission Electron Microscopy (TEM), Optical Microscopy and X-ray Photoelectron Spectroscopy (XPS). The new class of metallo-liposomes was used as a DNA delivery system and demonstrated to be effective for the transfection of pEGFP-N1 plasmid into HEK 293-T cells. Modular molecular

  6. Fungal inoculation and elevated CO2 mediate growth of Lolium mutiforum and Phytolacca americana, metal uptake, and metal bioavailability in metal-contaminated soil: evidence from DGT measurement.

    PubMed

    Song, Ningning; Wang, Fangli; Zhang, Changbo; Tang, Shirong; Guo, Junkang; Ju, Xuehai; Smith, Donald L

    2013-01-01

    Fungal inoculation and elevated CO2 may mediate plant growth and uptake of heavy metals, but little evidence from Diffusive Gradients in Thin-films (DGT) measurement has been obtained to characterize the process. Lolium mutiforum and Phytolacca americana were grown at ambient and elevated CO2 on naturally Cd and Pb contaminated soils inoculated with and without Trichoderma asperellum strain C3 or Penicillium chrysogenum strain D4, to investigate plant growth, metal uptake, and metal bioavailability responses. Fungal inoculation increased plant biomass and shoot/root Cd and Pb concentrations. Elevated CO2 significantly increased plants biomass, but decreased Cd and Pb concentrations in shoot/root to various extents, leading to a metal dilution phenomenon. Total Cd and Pb uptake by plants, and DGT-measured Cd and Pb concentrations in rhizosphere soils, were higher in all fungal inoculation and elevated CO2 treatments than control treatments, with the combined treatments having more influence than either treatment alone. Metal dilution phenomenon occurred because the increase in DGT-measured bioavailable metal pools in plant rhizosphere due to elevated CO2 was unable to match the increase in requirement for plant uptake of metals due to plant biomass increase.

  7. Metamaterials and Transformation Optics

    DTIC Science & Technology

    2014-01-31

    research is extend the analytical work in transformation optics (relating complex systems to simpler systems with the same spectral properties ) to... optics which via a transformation relates complex systems to simpler systems possessed of the same spectral properties . One good example is to be...mediated by the quantum fluctuations in electron density at the metal surfaces and are the most long ranged forces between nanoparticles . Fig. 1(a

  8. A Conserved Metal Binding Motif in the Bacillus subtilis Competence Protein ComFA Enhances Transformation.

    PubMed

    Chilton, Scott S; Falbel, Tanya G; Hromada, Susan; Burton, Briana M

    2017-08-01

    Genetic competence is a process in which cells are able to take up DNA from their environment, resulting in horizontal gene transfer, a major mechanism for generating diversity in bacteria. Many bacteria carry homologs of the central DNA uptake machinery that has been well characterized in Bacillus subtilis It has been postulated that the B. subtilis competence helicase ComFA belongs to the DEAD box family of helicases/translocases. Here, we made a series of mutants to analyze conserved amino acid motifs in several regions of B. subtilis ComFA. First, we confirmed that ComFA activity requires amino acid residues conserved among the DEAD box helicases, and second, we show that a zinc finger-like motif consisting of four cysteines is required for efficient transformation. Each cysteine in the motif is important, and mutation of at least two of the cysteines dramatically reduces transformation efficiency. Further, combining multiple cysteine mutations with the helicase mutations shows an additive phenotype. Our results suggest that the helicase and metal binding functions are two distinct activities important for ComFA function during transformation.IMPORTANCE ComFA is a highly conserved protein that has a role in DNA uptake during natural competence, a mechanism for horizontal gene transfer observed in many bacteria. Investigation of the details of the DNA uptake mechanism is important for understanding the ways in which bacteria gain new traits from their environment, such as drug resistance. To dissect the role of ComFA in the DNA uptake machinery, we introduced point mutations into several motifs in the protein sequence. We demonstrate that several amino acid motifs conserved among ComFA proteins are important for efficient transformation. This report is the first to demonstrate the functional requirement of an amino-terminal cysteine motif in ComFA. Copyright © 2017 American Society for Microbiology.

  9. Reversible electron-transfer reactions within a nanoscale metal oxide cage mediated by metallic substrates

    SciTech Connect

    Fleming, C.; Long, D. L.; McMillian, N.; Johnston, J.; Bovet N.; Dhanak, V.; Gadegaard, N.; Kogerler, P.; Cronin, L.; Kadodwala, M.

    2008-03-30

    Transition metal oxides exhibit a rich collection of electronic properties and have many practical applications in areas such as catalysis and ultra-high-density magnetic data storage. Therefore the development of switchable molecular transition metal oxides has potential for the engineering of single-molecule devices and nanoscale electronics. At present, the electronic properties of transition metal oxides can only be tailored through the irreversible introduction of dopant ions, modifying the electronic structure by either injecting electrons or core holes. Here we show that a molybdenum(VI) oxide 'polyoxometalate' molecular nanocluster containing two embedded redox agents is activated by a metallic surface and can reversibly interconvert between two electronic states. Upon thermal activation two electrons are ejected from the active sulphite anions and delocalized over the metal oxide cluster cage, switching it from a fully oxidized state to a two-electron reduced state along with the concomitant formation of an S-S bonding interaction between the two sulphur centres inside the cluster shell.

  10. Humic substances as a mediator for microbially catalyzed metal reduction

    USGS Publications Warehouse

    Lovley, D.R.; Fraga, J.L.; Blunt-Harris, E. L.; Hayes, L.A.; Phillips, E.J.P.; Coates, J.D.

    1998-01-01

    The potential for humic substances to serve as a terminal electron acceptor in microbial respiration and to function as an electron shuttle between Fe(III)-reducing microorganisms and insoluble Fe(III) oxides was investigated. The Fe(III)-reducing microorganism Geobacter metallireducens conserved energy to support growth from electron transport to humics as evidenced by continued oxidation of acetate to carbon dioxide after as many as nine transfers in a medium with acetate as the electron donor and soil humic acids as the electron acceptor. Growth of G. metallireducens with poorly crystalline Fe(III) oxide as the electron acceptor was greatly stimulated by the addition of as little as 100 ??M of the humics analog, anthraquinone-2,6-disulfonate. Other quinones investigated, including lawsone, menadione, and anthraquinone-2-sulfonate, also stimulated Fe(III) oxide reduction. A wide phylogenetic diversity of microorganisms capable of Fe(III) reduction were also able to transfer electrons to humics. Microorganisms which can not reduce Fe(III) could not reduce humics. Humics stimulated the reduction of structural Fe(III) in clay and the crystalline Fe(III) forms, goethite and hematite. These results demonstrate that electron shuttling between Fe(III)-reducing microorganisms and Fe(III) via humics not only accelerates the microbial reduction of poorly crystalline Fe(III) oxide, but also can facilitate the reduction of Fe(III) forms that are not typically reduced by microorganisms in the absence of humics. Addition of humic substances to enhance electron shuttling between Fe(III)-reducing microorganisms and Fe(III) oxides may be a useful strategy to stimulate the remediation of soils and sediments contaminated with organic or metal pollutants.

  11. Metal-mediated DNA base pairing: alternatives to hydrogen-bonded Watson-Crick base pairs.

    PubMed

    Takezawa, Yusuke; Shionoya, Mitsuhiko

    2012-12-18

    With its capacity to store and transfer the genetic information within a sequence of monomers, DNA forms its central role in chemical evolution through replication and amplification. This elegant behavior is largely based on highly specific molecular recognition between nucleobases through the specific hydrogen bonds in the Watson-Crick base pairing system. While the native base pairs have been amazingly sophisticated through the long history of evolution, synthetic chemists have devoted considerable efforts to create alternative base pairing systems in recent decades. Most of these new systems were designed based on the shape complementarity of the pairs or the rearrangement of hydrogen-bonding patterns. We wondered whether metal coordination could serve as an alternative driving force for DNA base pairing and why hydrogen bonding was selected on Earth in the course of molecular evolution. Therefore, we envisioned an alternative design strategy: we replaced hydrogen bonding with another important scheme in biological systems, metal-coordination bonding. In this Account, we provide an overview of the chemistry of metal-mediated base pairing including basic concepts, molecular design, characteristic structures and properties, and possible applications of DNA-based molecular systems. We describe several examples of artificial metal-mediated base pairs, such as Cu(2+)-mediated hydroxypyridone base pair, H-Cu(2+)-H (where H denotes a hydroxypyridone-bearing nucleoside), developed by us and other researchers. To design the metallo-base pairs we carefully chose appropriate combinations of ligand-bearing nucleosides and metal ions. As expected from their stronger bonding through metal coordination, DNA duplexes possessing metallo-base pairs exhibited higher thermal stability than natural hydrogen-bonded DNAs. Furthermore, we could also use metal-mediated base pairs to construct or induce other high-order structures. These features could lead to metal-responsive functional

  12. Promoting safety voice with safety-specific transformational leadership: the mediating role of two dimensions of trust.

    PubMed

    Conchie, Stacey M; Taylor, Paul J; Donald, Ian J

    2012-01-01

    Although safety-specific transformational leadership is known to encourage employee safety voice behaviors, less is known about what makes this style of leadership effective. We tested a model that links safety-specific transformational leadership to safety voice through various dimensions of trust. Data from 150 supervisor-employee dyads from the United Kingdom oil industry supported our predictions that the effects of safety-specific transformational leadership are sequentially mediated by affect-based trust beliefs and disclosure trust intentions. Moreover, we found that reliance trust intentions moderated the effect of disclosure: employees' disclosure intentions mediated the effects of affect-based trust on safety voice behaviors only when employees' intention to rely on their leader was moderate to high. These findings suggest that leaders seeking to encourage safety voice behaviors should go beyond "good reason" arguments and develop affective bonds with their employees.

  13. Long- and short-term temperature responses of microbially-mediated boreal soil organic matter transformations

    NASA Astrophysics Data System (ADS)

    Min, K.; Buckeridge, K. M.; Edwards, K. A.; Ziegler, S. E.; Billings, S. A.

    2015-12-01

    Microorganisms use exoenzymes to decay soil organic matter into assimilable substrates, some of which are transformed into CO2. Microbial CO2 efflux contributes up to 60% of soil respiration, a feature that can change with temperature due to altered exoenzyme activities (short-term) and microbial communities producing different exoenzymes (longer-term). Often, however, microbial temperature responses are masked by factors that also change with temperature in soil, making accurate projections of microbial CO2 efflux with warming challenging. Using soils along a natural climate gradient similar in most respects except for temperature regime (Newfoundland Labrador Boreal Ecosystem Latitudinal Transect), we investigated short-vs. long-term temperature responses of microbially-mediated organic matter transformations. While incubating soils at 5, 15, and 25°C for 84 days, we measured exoenzyme activities, CO2 efflux rates and biomass, and extracted DNA at multiple times. We hypothesized that short-term, temperature-induced increases in exoenzyme activities and CO2 losses would be smaller in soils from warmer regions, because microbes presumably adapted to warmer regions should use assimilable substrates more efficiently and thus produce exoenzymes at a lower rate. While incubation temperature generally induced greater exoenzyme activities (p<0.001), exoenzymes' temperature responses depended on enzymes and regions (p<0.001). Rate of CO2 efflux was affected by incubation temperature (P<0.001), but not by region. Microbial biomass and DNA sequencing will reveal how microbial community abundance and composition change with short-vs. longer-term temperature change. Though short-term microbial responses to temperature suggest higher CO2 efflux and thus lower efficiency of resource use with warming, longer-term adaptations of microbial communities to warmer climates remain unknown; this work helps fill that knowledge gap.

  14. 4-Hydroxyestradiol induces mammary epithelial cell transformation through Nrf2-mediated heme oxygenase-1 overexpression

    PubMed Central

    Park, Sin-Aye; Lee, Mee-Hyun; Na, Hye-Kyung; Surh, Young-Joon

    2017-01-01

    Estrogen (17β-estradiol, E2) undergoes oxidative metabolism by CYP1B1 to form 4-hydroxyestradiol (4-OHE2), a putative carcinogenic metabolite of estrogen. Our previous study showed that 4-OHE2-induced production of reactive oxygen species contributed to neoplastic transformation of human breast epithelial (MCF-10A) cells. In this study, 4-OHE2, but not E2, increased the expression of heme oxygenase-1 (HO-1), a sensor and regulator of oxidative stress, in MCF-10A cells. Silencing the HO-1 gene in MCF-10A cells suppressed 4-OHE2-induced cell proliferation and transformation. In addition, subcutaneous administration of 4-OHE2 markedly enhanced the growth of the MDA-MB-231 human breast cancer xenografts, which was retarded by zinc protoporphyrin, a pharmacological inhibitor of HO-1. 4-OHE2-induced HO-1 expression was mediated by NF-E2-related factor 2 (Nrf2). We speculate that an electrophilic quinone formed as a consequence of oxidation of 4-OHE2 binds directly to Kelch-like ECH-associated protein 1 (Keap1), an inhibitory protein that sequesters Nrf2 in the cytoplasm. This will diminish association between Nrf2 and Keap1. 4-OHE2 failed to interrupt the interaction between Keap1 and Nrf2 and to induce HO-1 expression in Keap1-C273S or C288S mutant cells. Lano-LC-ESI-MS/MS analysis in MCF-10A-Keap1-WT cells which were treated with 4-OHE2 revealed that the peptide fragment containing Cys288 gained a molecular mass of 287.15 Da, equivalent to the addition of a single molecule of 4-OHE2-derived ortho-quinones. PMID:27438141

  15. Spatial variance of POPs and heavy metals in transformer oil-contaminated soil around Tamil Nadu.

    PubMed

    Murugan, Karuvelan; Vasudevan, Namasivayam

    2017-09-05

    The persistent organic pollutants in the environment are one of the global issues to their unregulated disposal and informal recycling. This study investigates the contamination of soil with polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), phenolic compounds and heavy metals via spillage of transformer oil (TO). Fresh TO (FTO), used TO (UTO) and soil samples were analysed using GC-MS to confirm the presence of 8 PCB congeners, 16 PAHs and 24 types of phenolic compounds and using inductively coupled plasma optical emission spectrometry to confirm the presence of 7 heavy metals. The chromatographic analysis revealed the levels of mono-, di-, tri-, tetra-, penta-, hexa-, hepta- and octachlorobiphenyls in FTO to be 5.63, 25.24, 0.195, 0.185, 2.169, 1.023 and 5.28 mg/L and the level of mono-, di-, tri-, tetra-, penta- and hexachlorobiphenyls in UTO to be 0.27, 1.21, 1.31, 0.80, 1.77 and 3.90 mg/L. Analysis of soil from 10 different TO-contaminated sites showed the presence of PCBs, PAHs, phenolic compounds and heavy metals in the concentration range of 0.53-42.87 mg/kg, 3.19-246.6 μg/kg, 0.01-4086.45 μg/kg and 1.0-401.3 mg/kg, respectively. The variation in the concentration of these compounds and heavy metals among different sampling sites was determined using principal component analysis (PCA), metric multi-dimensional scaling (MDS) and Bray-Curtis cluster analysis (Bu-CA). The toxicity equivalence factor and the mechanism involved in the disruption of endocrine system are discussed. Thus, this study exemplifies the need for complete ban of PCB-containing TOs in developing countries and urges the need for technology for the disposal of TO.

  16. Valproic acid overcomes transforming growth factor-β-mediated sorafenib resistance in hepatocellular carcinoma

    PubMed Central

    Matsuda, Yasunobu; Wakai, Toshifumi; Kubota, Masayuki; Osawa, Mami; Hirose, Yuki; Sakata, Jun; Kobayashi, Takashi; Fujimaki, Shun; Takamura, Masaaki; Yamagiwa, Satoshi; Aoyagi, Yutaka

    2014-01-01

    Sorafenib is a multi-kinase inhibitor approved for hepatocellular carcinoma, but rarely causes tumor regression in patients with chronic liver diseases. To investigate whether growth factor-mediated signaling is involved in sorafenib resistance, HepG2 and PLC/PRF/5 hepatoma cells were exposed to epidermal growth factor (EGF), hepatocyte growth factor (HGF) or transforming growth factor-β (TGF-β) prior to treatment with sorafenib. Furthermore, to identify an effective combination treatment with sorafenib, growth factor-sensitized cells were treated with sorafenib alone or in combination with celecoxib, lovastatin or valproic acid (VPA). Trypan blue staining and Annexin V assays showed that the cytotoxic effect of sorafenib was inhibited by 15-54% in cells sensitized to TGF-β (P<0.05). Western blotting analysis showed that TGF-β significantly activated extracellular signal-regulated kinase (ERK)-mediated AKT signaling, and sorafenib failed to suppress both ERK and AKT in TGF-β-sensitized cells. The decreased anti-tumor effect of sorafenib was rescued by chemical inhibition of ERK and AKT. When TGF-β-sensitized cells were treated with sorafenib plus VPA, the levels of phosphorylated ERK and AKT were considerably suppressed and the numbers of dead cells were increased by 3.7-5.7-fold compared with those exposed to sorafenib alone (P<0.05). Moreover, low dose sorafenib-induced cell migration was effectively suppressed by combination treatment with sorafenib and VPA. Collectively, TGF-β/ERK/AKT signaling might play a critical role in sorafenib resistance in hepatoma cells, and combination treatment with VPA may be effective against this drug resistance. PMID:24817927

  17. Gremlin is a downstream profibrotic mediator of transforming growth factor-beta in cultured renal cells.

    PubMed

    Rodrigues-Diez, Raquel; Lavoz, Carolina; Carvajal, Gisselle; Rayego-Mateos, Sandra; Rodrigues Diez, Raul R; Ortiz, Alberto; Egido, Jesús; Mezzano, Sergio; Ruiz-Ortega, Marta

    2012-01-01

    Chronic kidney disease is characterized by accumulation of extracellular matrix in the tubulointerstitial area. Fibroblasts are the main matrix-producing cells. One source of activated fibroblasts is the epithelial mesenchymal transition (EMT). In cultured tubular epithelial cells, transforming growth factor-β (TGF-β1) induced Gremlin production associated with EMT phenotypic changes, and therefore Gremlin has been proposed as a downstream TGF-β1 mediator. Gremlin is a developmental gene upregulated in chronic kidney diseases associated with matrix accumulation, but its direct role in the modulation of renal fibrosis and its relation with TGF-β has not been investigated. Murine renal fibroblasts and human tubular epithelial cells were studied. Renal fibrosis was determined by evaluation of key profibrotic factors, extracellular matrix proteins (ECM) and EMT markers by Western blot/confocal microscopy or real-time PCR. Endogenous Gremlin was targeted with small interfering RNA. In murine fibroblasts, stimulation with recombinant Gremlin upregulated profibrotic genes, such as TGF-β1, and augmented the production of ECM proteins, including type I collagen. The blockade of endogenous Gremlin with small interfering RNA inhibited TGF-β1-induced ECM upregulation. In tubular epithelial cells Gremlin also increased profibrotic genes and caused EMT changes: phenotypic modulation to myofibroblast-like morphology, loss of epithelial markers and in-duction of mesenchymal markers. Moreover, Gremlin gene silencing inhibited TGF-β1-induced EMT changes. Gremlin directly activates profibrotic events in cul-tured renal fibroblasts and tubular epithelial cells. Moreover, endogenous Gremlin blockade inhibited TGF-β-mediated matrix production and EMT, suggesting that Gremlin could be a novel therapeutic target for renal fibrosis. Copyright © 2013 S. Karger AG, Basel.

  18. Magnetostructural martensitic transformations with large volume changes and magneto-strains in all-d-metal Heusler alloys

    NASA Astrophysics Data System (ADS)

    Wei, Z. Y.; Liu, E. K.; Li, Y.; Han, X. L.; Du, Z. W.; Luo, H. Z.; Liu, G. D.; Xi, X. K.; Zhang, H. W.; Wang, W. H.; Wu, G. H.

    2016-08-01

    The all-d-metal Mn2-based Heusler ferromagnetic shape memory alloys Mn50Ni40-xCoxTi10 (x = 8 and 9.5) are realized. With a generic comparison between d-metal Ti and main-group elements in lowering the transformation temperature, the magnetostructural martensitic transformations are established by further introducing Co to produce local ferromagnetic Mn-Co-Mn configurations. A 5-fold modulation and (3, -2) stacking of [00 10] of martensite are determined by X-ray diffraction and HRTEM analysis. Based on the transformation, a large magneto-strain of 6900 ppm and a large volume change of -2.54% are observed in polycrystalline samples, which makes the all-d-metal magnetic martensitic alloys of interest for magnetic/pressure multi-field driven applications.

  19. Magnetostructural martensitic transformations with large volume changes and magneto-strains in all-d-metal Heusler alloys

    SciTech Connect

    Wei, Z. Y.; Liu, E. K. Xi, X. K.; Zhang, H. W.; Wang, W. H.; Wu, G. H.; Li, Y.; Han, X. L.; Du, Z. W.; Luo, H. Z.; Liu, G. D.

    2016-08-15

    The all-d-metal Mn{sub 2}-based Heusler ferromagnetic shape memory alloys Mn{sub 50}Ni{sub 40−x}Co{sub x}Ti{sub 10} (x = 8 and 9.5) are realized. With a generic comparison between d-metal Ti and main-group elements in lowering the transformation temperature, the magnetostructural martensitic transformations are established by further introducing Co to produce local ferromagnetic Mn-Co-Mn configurations. A 5-fold modulation and (3, −2) stacking of [00 10] of martensite are determined by X-ray diffraction and HRTEM analysis. Based on the transformation, a large magneto-strain of 6900 ppm and a large volume change of −2.54% are observed in polycrystalline samples, which makes the all-d-metal magnetic martensitic alloys of interest for magnetic/pressure multi-field driven applications.

  20. RGS6 Suppresses Ras-induced Cellular Transformation by Facilitating Tip60-mediated Dnmt1 Degradation and Promoting Apoptosis

    PubMed Central

    Huang, Jie; Stewart, Adele; Maity, Biswanath; Hagen, Jussara; Fagan, Rebecca L.; Yang, Jianqi; Quelle, Dawn E.; Brenner, Charles; Fisher, Rory A.

    2014-01-01

    The RAS protooncogene plays a central role in regulation of cell proliferation, and point mutations leading to oncogenic activation of Ras occur in a large number of human cancers. Silencing of tumor suppressor genes by DNA methyltransferase 1 (Dnmt1) is essential for oncogenic cellular transformation by Ras, and Dnmt1 is over-expressed in numerous human cancers. Here we provide new evidence that the pleiotropic Regulator of G protein Signaling (RGS) family member RGS6 suppresses Ras-induced cellular transformation by facilitating Tip60-mediated degradation of Dmnt1 and promoting apoptosis. Employing mouse embryonic fibroblasts (MEFs) from wild type (WT) and RGS6−/− mice, we found that oncogenic Ras induced up-regulation of RGS6, which in turn blocked Ras-induced cellular transformation. RGS6 functions to suppress cellular transformation in response to oncogenic Ras by down regulating Dnmt1 protein expression leading to inhibition of Dnmt1-mediated anti-apoptotic activity. Further experiments showed that RGS6 functions as a scaffolding protein for both Dnmt1 and Tip60 and is required for Tip60-mediated acetylation of Dnmt1 and subsequent Dnmt1 ubiquitylation and degradation. The RGS domain of RGS6, known only for its GAP activity toward Gα subunits, was sufficient to mediate Tip60 association with RGS6. This work demonstrates a novel signaling action for RGS6 in negative regulation of oncogene-induced transformation and provides new insights into our understanding of the mechanisms underlying Ras-induced oncogenic transformation and regulation of Dnmt1 expression. Importantly, these findings identify RGS6 as an essential cellular defender against oncogenic stress and a potential therapeutic target for developing new cancer treatments. PMID:23995786

  1. Does self-efficacy mediate the relationship between transformational leadership behaviours and healthcare workers' sleep quality? A longitudinal study.

    PubMed

    Munir, Fehmidah; Nielsen, Karina

    2009-09-01

    This paper is a report of a study conducted to investigate the longitudinal relationship between transformational leadership behaviours and employees' sleep quality, and the mediating effects of self-efficacy. Although there is evidence for the influential role of transformational leadership on health outcomes, researchers have used either attitude outcomes (e.g. job satisfaction) or softer health measures, such as general well-being. Specific measures of well-being such as sleep quality have not been used, despite its association with working conditions. A longitudinal design was used to collect data from Danish healthcare workers at time 1 in 2005 (n = 447) and 18 months later at time 2 in 2007 (n = 274). Structural equation modelling was used to investigate the relationships between transformational leadership, self-efficacy and sleep quality at both time points independently (cross-sectionally) and longitudinally. For all constructs, time 2 measures were influenced by the baseline level. Direct relationships between transformational leadership and sleep quality were found. This relationship was negative cross-sectionally at both time points, but positive between baseline and follow-up. The relationship between leadership and employees' sleep quality was not mediated by employees' self-efficacy. Our results indicate that training managers in transformational leadership behaviours may have a positive impact on healthcare workers' health over time. However, more research is needed to examine the mechanisms by which transformational leadership brings about improved sleep quality; self-efficacy was not found to be the explanation.

  2. Agrobacterium tumefaciens-mediated transformation in the entomopathogenic fungus Lecanicillium lecanii and development of benzimidazole fungicide resistant strains.

    PubMed

    Zhang, Yan-Jun; Zhao, Jin-Jin; Xie, Ming; Peng, De-Liang

    2014-10-01

    Lecanicillium lecanii has been used in the biological control of several insects in agricultural practice. Since the gene manipulation tools for this entomopathogenic fungus have not been sufficiently developed, Agrobacterium tumefaciens-mediated transformation (ATMT) in L. lecanii was investigated in this study, using the wild-type isolate FZ9906 as a progenitor strain and the hygromycin B resistance (hph) gene as a selection marker. Furthermore, a field carbendazim-resistant (mrt) gene from Botrytis cinerea was expressed in L. lecanii FZ9906 via the ATMT system. The results revealed that the frequency of transformation surpassed 25transformants/10(6) conidia, most of the putative transformants contained a single copy of T-DNA, and the T-DNA inserts were stably inherited after five generations. All putative transformants had indistinguishable biological characteristics relative to the wild-type strain, excepting two transformants with altered growth habits or virulence. Moreover, the resistance of the putative transformants to carbendazim (MBC) was improved, and the highest one was 380-fold higher than the wild-type strain. In conclusion, ATMT is an effective and suitable system for L. lecanii transformation, and will be a useful tool for the basic and application research of gene functions and gene modifications of this strain.

  3. The development of coil short circuits when transformer windings become contaminated with metal-containing colloidal particles

    SciTech Connect

    L'vov, S. Yu.; Lyut'ko, E. O.; Bondareva, V. N.; Komarov, V. B.; L'vov, Yu. N.

    2012-01-15

    The radiational-thermal development of coil short circuits due to the action of partial discharges of the first kind when the windings of transformers, autotransformers and shunting reactors become contaminated with metal-containing colloidal particles, formed in the transformer oil as a result of the interaction of the oil with the constructional materials (the copper of the windings, the iron of the tank, the core etc.) is considered. Acriterion of dangerous contamination of the coil insulation of the windings by metal-containing colloidal particles is proposed, namely, 3% of the mass content of copper and iron in it, which, if exceeded, may serve as a basis for recognizing the state of transformers, autotransformers and shunting reactors at a voltage of 110 kV and above the limit. It is shown that filters for continuously cleaning the oil play a considerable role in prolonging the life of transformer equipment.

  4. An approach to crystallizing proteins by metal-mediated synthetic symmetrization

    SciTech Connect

    Laganowsky, Arthur; Zhao, Minglei; Soriaga, Angela B.; Sawaya, Michael R.; Cascio, Duilio; Yeates, Todd O.

    2011-12-09

    Combining the concepts of synthetic symmetrization with the approach of engineering metal-binding sites, we have developed a new crystallization methodology termed metal-mediated synthetic symmetrization. In this method, pairs of histidine or cysteine mutations are introduced on the surface of target proteins, generating crystal lattice contacts or oligomeric assemblies upon coordination with metal. Metal-mediated synthetic symmetrization greatly expands the packing and oligomeric assembly possibilities of target proteins, thereby increasing the chances of growing diffraction-quality crystals. To demonstrate this method, we designed various T4 lysozyme (T4L) and maltose-binding protein (MBP) mutants and cocrystallized them with one of three metal ions: copper (Cu2+), nickel (Ni2+), or zinc (Zn2+). The approach resulted in 16 new crystal structures-eight for T4L and eight for MBP-displaying a variety of oligomeric assemblies and packing modes, representing in total 13 new and distinct crystal forms for these proteins. We discuss the potential utility of the method for crystallizing target proteins of unknown structure by engineering in pairs of histidine or cysteine residues. As an alternate strategy, we propose that the varied crystallization-prone forms of T4L or MBP engineered in this work could be used as crystallization chaperones, by fusing them genetically to target proteins of interest.

  5. Radon transform based automatic metal artefacts generation for 3D threat image projection

    NASA Astrophysics Data System (ADS)

    Megherbi, Najla; Breckon, Toby P.; Flitton, Greg T.; Mouton, Andre

    2013-10-01

    Threat Image Projection (TIP) plays an important role in aviation security. In order to evaluate human security screeners in determining threats, TIP systems project images of realistic threat items into the images of the passenger baggage being scanned. In this proof of concept paper, we propose a 3D TIP method which can be integrated within new 3D Computed Tomography (CT) screening systems. In order to make the threat items appear as if they were genuinely located in the scanned bag, appropriate CT metal artefacts are generated in the resulting TIP images according to the scan orientation, the passenger bag content and the material of the inserted threat items. This process is performed in the projection domain using a novel methodology based on the Radon Transform. The obtained results using challenging 3D CT baggage images are very promising in terms of plausibility and realism.

  6. Phase Transformations and Metallization of Magnesium Oxide at High Pressure and Temperature

    NASA Astrophysics Data System (ADS)

    McWilliams, R. Stewart; Spaulding, Dylan K.; Eggert, Jon H.; Celliers, Peter M.; Hicks, Damien G.; Smith, Raymond F.; Collins, Gilbert W.; Jeanloz, Raymond

    2012-12-01

    Magnesium oxide (MgO) is representative of the rocky materials comprising the mantles of terrestrial planets, such that its properties at high temperatures and pressures reflect the nature of planetary interiors. Shock-compression experiments on MgO to pressures of 1.4 terapascals (TPa) reveal a sequence of two phase transformations: from B1 (sodium chloride) to B2 (cesium chloride) crystal structures above 0.36 TPa, and from electrically insulating solid to metallic liquid above 0.60 TPa. The transitions exhibit large latent heats that are likely to affect the structure and evolution of super-Earths. Together with data on other oxide liquids, we conclude that magmas deep inside terrestrial planets can be electrically conductive, enabling magnetic field-producing dynamo action within oxide-rich regions and blurring the distinction between planetary mantles and cores.

  7. Plastic deformation of metallic glasses: Size of shear transformation zones from molecular dynamics simulations

    NASA Astrophysics Data System (ADS)

    Zink, Mareike; Samwer, K.; Johnson, W. L.; Mayr, S. G.

    2006-05-01

    Plastic deformation in metallic glasses well below their glass transition temperatures Tg occurs spatially heterogeneously within highly localized regions, termed shear transformation zones (STZs). Yet, their size and the number of atoms involved in a local shear event, remains greatly unclear. With the help of classical molecular dynamics (MD) computer simulations on plastic deformation of the model glass CuTi during pure shearing, we address this issue by evaluating correlations in atomic-scale plastic displacements, viz. the displacement correlation function. From the correlation length, a universal diameter of about 15Å , or, equivalently, approximately 120 atoms is derived for a variety of conditions, such as variable strains, strain rates, temperatures, and boundary conditions. Our findings are consistent with a recent model proposed by Johnson and Samwer [Phys. Rev. Lett. 95, 195501 (2005)].

  8. Reversible transformations of silver oxide and metallic silver nanoparticles inside SiO{sub 2} films

    SciTech Connect

    Pal, Sudipto; De, Goutam

    2009-02-04

    Reversible transformation of silver oxide and metallic nanoparticles inside a relatively porous silica film has been established. Annealing of Ag-doped films in oxidizing (air) atmosphere at 450 deg. C yielded colorless films containing AgO{sub x}. These films were turned yellow when heated in H{sub 2}-N{sub 2} (reducing atmosphere) due to the formation of Ag nanoparticles. This yellow coloration (due to nano Ag{sup 0}) and bleaching (conversion of Ag{sup 0} {yields} Ag{sup +}) are reversible. Optical and photoluminescence spectra are well consistent with this coloration and bleaching. The soaking test of the air-annealed film in Na{sub 2}S{sub 2}O{sub 3} solution supports the presence of Ag{sup +}. Grazing incidence X-ray diffraction and transmission electron microscopy studies reveal the formation of Ag-oxides and Ag nanoparticles in the oxidized and reduced films, respectively.

  9. Composition mediated serration dynamics in Zr-based bulk metallic glasses

    NASA Astrophysics Data System (ADS)

    Wang, Z.; Qiao, J. W.; Tian, H.; Sun, B. A.; Wang, B. C.; Xu, B. S.; Chen, M. W.

    2015-11-01

    The composition mediated serration dynamics in Zr-based bulk metallic glasses (BMGs) is investigated by statistics analyses of the elastic-energy density, and free volumes during shear-banding are beneficial to understand serrated-flow behavior. The amplitude and elastic-energy density display a gradually increasing and then decreasing trend with increasing the content of Zr. It is based on the free-volume theory describing the atomic-level structure of ternary Zr-Cu-Al BMGs. The good agreement between the molecular dynamics simulation and experimental results provides evidence for the variation of free volumes as the elementary mechanism of composition mediated serration dynamics.

  10. Composition mediated serration dynamics in Zr-based bulk metallic glasses

    SciTech Connect

    Wang, Z.; Qiao, J. W. E-mail: mwchen@wpi-aimr.tohoku.ac.jp; Wang, B. C.; Xu, B. S.; Tian, H.; Sun, B. A.; Chen, M. W. E-mail: mwchen@wpi-aimr.tohoku.ac.jp

    2015-11-16

    The composition mediated serration dynamics in Zr-based bulk metallic glasses (BMGs) is investigated by statistics analyses of the elastic-energy density, and free volumes during shear-banding are beneficial to understand serrated-flow behavior. The amplitude and elastic-energy density display a gradually increasing and then decreasing trend with increasing the content of Zr. It is based on the free-volume theory describing the atomic-level structure of ternary Zr-Cu-Al BMGs. The good agreement between the molecular dynamics simulation and experimental results provides evidence for the variation of free volumes as the elementary mechanism of composition mediated serration dynamics.

  11. Molecular Recognition-Mediated Transformation of Single-Chain Polymer Nanoparticles into Crosslinked Polymer Films.

    PubMed

    Mahon, Clare S; McGurk, Christopher J; Watson, Scott M D; Fascione, Martin A; Sakonsinsiri, Chadamas; Turnbull, W Bruce; Fulton, David A

    2017-08-14

    We describe single-chain polymer nanoparticles (SCNPs) possessing intramolecular dynamic covalent crosslinks that can transform into polymer films through a molecular recognition-mediated crosslinking process. The SCNPs utilise molecular recognition with surface-immobilised proteins to concentrate upon a substrate, bringing the SCNPs into close spatial proximity with one another and allowing their dynamic covalent crosslinkers to undergo intra- to interpolymer chain crosslinking leading to the formation of polymeric film. SCNPs must possess both the capacity for specific molecular recognition and a dynamic nature to their intramolecular crosslinkers to form polymer films, and an investigation of the initial phase of film formation indicates it proceeds from features which form upon the surface then grow predominantly in the xy directions. This approach to polymer film formation presents a potential method to "wrap" surfaces displaying molecular recognition motifs-which could potentially include viral, cellular and bacterial surfaces or artificial surfaces displaying multivalent recognition motifs-within a layer of polymer film. © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  12. Adaptation or Malignant Transformation: The Two Faces of Epigenetically Mediated Response to Stress

    PubMed Central

    Zoldoš, Vlatka

    2013-01-01

    Adaptive response to stress is a fundamental property of living systems. At the cellular level, many different types of stress elicit an essentially limited repertoire of adaptive responses. Epigenetic changes are the main mechanism for medium- to long-term adaptation to accumulated (intense, long-term, or repeated) stress. We propose the adaptive deregulation of the epigenome in response to stress (ADERS) hypothesis which assumes that the unspecific adaptive stress response grows stronger with the increasing stress level, epigenetically activating response gene clusters while progressively deregulating other cellular processes. The balance between the unspecific adaptive response and the general epigenetic deregulation is critical because a strong response can lead to pathology, particularly to malignant transformation. The main idea of our hypothesis is the continuum traversed by a cell subjected to accumulated stress, which lies between an unspecific adaptive response and pathological deregulation—the two extremes sharing the same underlying cause, which is a manifestation of a unified epigenetically mediated adaptive response to stress. The evolutionary potential of epigenetic regulation in multigenerational adaptation is speculatively discussed in the light of neo-Lamarckism. Finally, an approach to testing the proposed hypothesis is presented, relying on either the publicly available datasets or on conducting new experiments. PMID:24187667

  13. Adaptation or malignant transformation: the two faces of epigenetically mediated response to stress.

    PubMed

    Vojta, Aleksandar; Zoldoš, Vlatka

    2013-01-01

    Adaptive response to stress is a fundamental property of living systems. At the cellular level, many different types of stress elicit an essentially limited repertoire of adaptive responses. Epigenetic changes are the main mechanism for medium- to long-term adaptation to accumulated (intense, long-term, or repeated) stress. We propose the adaptive deregulation of the epigenome in response to stress (ADERS) hypothesis which assumes that the unspecific adaptive stress response grows stronger with the increasing stress level, epigenetically activating response gene clusters while progressively deregulating other cellular processes. The balance between the unspecific adaptive response and the general epigenetic deregulation is critical because a strong response can lead to pathology, particularly to malignant transformation. The main idea of our hypothesis is the continuum traversed by a cell subjected to accumulated stress, which lies between an unspecific adaptive response and pathological deregulation--the two extremes sharing the same underlying cause, which is a manifestation of a unified epigenetically mediated adaptive response to stress. The evolutionary potential of epigenetic regulation in multigenerational adaptation is speculatively discussed in the light of neo-Lamarckism. Finally, an approach to testing the proposed hypothesis is presented, relying on either the publicly available datasets or on conducting new experiments.

  14. Riboflavin-mediated RDX transformation in the presence of Shewanella putrefaciens CN32 and lepidocrocite.

    PubMed

    Bae, Sungjun; Lee, Yoonhwa; Kwon, Man Jae; Lee, Woojin

    2014-06-15

    The potential of riboflavin for the reductive degradation of a cyclic nitramine, hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), was investigated in the presence of lepidocrocite and/or Shewanella putrefaciens CN32. RDX reduction by CN32 alone or CN32 with lepidocrocite was insignificant, while 110 μM RDX was completely reduced by CN32 with riboflavin in 78 h. The transformation products identified included nitroso metabolites, formaldehyde, and ammonium, indicating the ring cleavage of RDX. UV and visible light analysis revealed that riboflavin was microbially reduced by CN32, and that the reduced riboflavin was linked to the complete degradation of RDX. In the presence of both CN32 and lepidocrocite (γ-FeOOH), 100 μM-riboflavin increased the rate and extent of Fe(II) production as well as RDX reduction. An abiotic study also showed that Fe(II)-riboflavin complex, and Fe(II) adsorbed on lepidocrocite, reduced RDX by 48% and 21%, respectively. The findings in this study suggest that riboflavin-mediated RDX degradation pathways in subsurface environments are diverse and complex. However, riboflavin, either from bacteria or exogenous sources, can significantly increase RDX degradation. This will provide a sustainable clean-up option for explosive-contaminated subsurface environments. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Microbial- and thiosulfate-mediated dissolution of mercury sulfide minerals and transformation to gaseous mercury.

    PubMed

    Vázquez-Rodríguez, Adiari I; Hansel, Colleen M; Zhang, Tong; Lamborg, Carl H; Santelli, Cara M; Webb, Samuel M; Brooks, Scott C

    2015-01-01

    Mercury (Hg) is a toxic heavy metal that poses significant environmental and human health risks. Soils and sediments, where Hg can exist as the Hg sulfide mineral metacinnabar (β-HgS), represent major Hg reservoirs in aquatic environments. Metacinnabar has historically been considered a sink for Hg in all but severely acidic environments, and thus disregarded as a potential source of Hg back to aqueous or gaseous pools. Here, we conducted a combination of field and laboratory incubations to identify the potential for metacinnabar as a source of dissolved Hg within near neutral pH environments and the underpinning (a)biotic mechanisms at play. We show that the abundant and widespread sulfur-oxidizing bacteria of the genus Thiobacillus extensively colonized metacinnabar chips incubated within aerobic, near neutral pH creek sediments. Laboratory incubations of axenic Thiobacillus thioparus cultures led to the release of metacinnabar-hosted Hg(II) and subsequent volatilization to Hg(0). This dissolution and volatilization was greatly enhanced in the presence of thiosulfate, which served a dual role by enhancing HgS dissolution through Hg complexation and providing an additional metabolic substrate for Thiobacillus. These findings reveal a new coupled abiotic-biotic pathway for the transformation of metacinnabar-bound Hg(II) to Hg(0), while expanding the sulfide substrates available for neutrophilic chemosynthetic bacteria to Hg-laden sulfides. They also point to mineral-hosted Hg as an underappreciated source of gaseous elemental Hg to the environment.

  16. Microbial- and thiosulfate-mediated dissolution of mercury sulfide minerals and transformation to gaseous mercury

    PubMed Central

    Vázquez-Rodríguez, Adiari I.; Hansel, Colleen M.; Zhang, Tong; Lamborg, Carl H.; Santelli, Cara M.; Webb, Samuel M.; Brooks, Scott C.

    2015-01-01

    Mercury (Hg) is a toxic heavy metal that poses significant environmental and human health risks. Soils and sediments, where Hg can exist as the Hg sulfide mineral metacinnabar (β-HgS), represent major Hg reservoirs in aquatic environments. Metacinnabar has historically been considered a sink for Hg in all but severely acidic environments, and thus disregarded as a potential source of Hg back to aqueous or gaseous pools. Here, we conducted a combination of field and laboratory incubations to identify the potential for metacinnabar as a source of dissolved Hg within near neutral pH environments and the underpinning (a)biotic mechanisms at play. We show that the abundant and widespread sulfur-oxidizing bacteria of the genus Thiobacillus extensively colonized metacinnabar chips incubated within aerobic, near neutral pH creek sediments. Laboratory incubations of axenic Thiobacillus thioparus cultures led to the release of metacinnabar-hosted Hg(II) and subsequent volatilization to Hg(0). This dissolution and volatilization was greatly enhanced in the presence of thiosulfate, which served a dual role by enhancing HgS dissolution through Hg complexation and providing an additional metabolic substrate for Thiobacillus. These findings reveal a new coupled abiotic-biotic pathway for the transformation of metacinnabar-bound Hg(II) to Hg(0), while expanding the sulfide substrates available for neutrophilic chemosynthetic bacteria to Hg-laden sulfides. They also point to mineral-hosted Hg as an underappreciated source of gaseous elemental Hg to the environment. PMID:26157421

  17. A theoretical study of the omega-phase transformation in metals

    NASA Astrophysics Data System (ADS)

    Sanati, Mahdi

    I have studied the formation of o-phase from electronic and mesoscopic (domain wall) points of view. To study the formation of domain walls, I have extended the Landau model of Cook for the o-phase transition by including a spatial gradient (Ginzburg) term of the scalar order parameter. In general, the Landau free energy is an asymmetric double-well potential. From the variational derivative of the total free energy I obtained a static equilibrium condition. By solving this equation for different physical parameters and boundary conditions, I obtained different quasi-one-dimensional soliton-like solutions. These solutions correspond to three different types of domain walls between the o-phase and the beta-matrix. These results are used to model the formation of the o-phase in bcc Ti. Canonical band model and first principles calculations confirmed the instability of the bcc-phase of group III and IV transition metals with respect to the o-phase transformation. I showed that the d-electron density is the controlling parameter for this type of the transformation. Also the possibility of formation of the o-phase for rare earth metals is discussed. First-principles full-potential linear muffin-tin orbital method (FPLMTO) calculations are performed for o-type displacement of the atoms to study the formation of the o-phase in TiAl and Ti 3Al2Nb alloys. The results of my calculations showed an instability in ordered B2 TiAl structure with respect to the o-phase when one third of the Al atoms are replaced by Nb atoms. These phenomena are explained, first by symmetry arguments; then a pair potential model is used to illustrate this instability based on interactions between different pair of atoms derived from the electronic structure. In addition, importance of the atomic arrangements on the structural stability of the Ti3Al2 Nb system is discussed.

  18. Engineering island-chain silicon nanowires via a droplet mediated Plateau-Rayleigh transformation

    PubMed Central

    Xue, Zhaoguo; Xu, Mingkun; Zhao, Yaolong; Wang, Jimmy; Jiang, Xiaofan; Yu, Linwei; Wang, Junzhuan; Xu, Jun; Shi, Yi; Chen, Kunji; Roca i Cabarrocas, Pere

    2016-01-01

    The ability to program highly modulated morphology upon silicon nanowires (SiNWs) has been fundamental to explore new phononic and electronic functionalities. We here exploit a nanoscale locomotion of metal droplets to demonstrate a large and readily controllable morphology engineering of crystalline SiNWs, from straight ones into continuous or discrete island-chains, at temperature <350 °C. This has been accomplished via a tin (Sn) droplet mediated in-plane growth where amorphous Si thin film is consumed as precursor to produce crystalline SiNWs. Thanks to a significant interface-stretching effect, a periodic Plateau-Rayleigh instability oscillation can be stimulated in the liquid Sn droplet, and the temporal oscillation of the Sn droplets is translated faithfully, via the deformable liquid/solid deposition interface, into regular spatial modulation upon the SiNWs. Combined with a unique self-alignment and positioning capability, this new strategy could enable a rational design and single-run fabrication of a wide variety of nanowire-based optoelectronic devices. PMID:27682161

  19. Engineering island-chain silicon nanowires via a droplet mediated Plateau-Rayleigh transformation

    NASA Astrophysics Data System (ADS)

    Xue, Zhaoguo; Xu, Mingkun; Zhao, Yaolong; Wang, Jimmy; Jiang, Xiaofan; Yu, Linwei; Wang, Junzhuan; Xu, Jun; Shi, Yi; Chen, Kunji; Roca I Cabarrocas, Pere

    2016-09-01

    The ability to program highly modulated morphology upon silicon nanowires (SiNWs) has been fundamental to explore new phononic and electronic functionalities. We here exploit a nanoscale locomotion of metal droplets to demonstrate a large and readily controllable morphology engineering of crystalline SiNWs, from straight ones into continuous or discrete island-chains, at temperature <350 °C. This has been accomplished via a tin (Sn) droplet mediated in-plane growth where amorphous Si thin film is consumed as precursor to produce crystalline SiNWs. Thanks to a significant interface-stretching effect, a periodic Plateau-Rayleigh instability oscillation can be stimulated in the liquid Sn droplet, and the temporal oscillation of the Sn droplets is translated faithfully, via the deformable liquid/solid deposition interface, into regular spatial modulation upon the SiNWs. Combined with a unique self-alignment and positioning capability, this new strategy could enable a rational design and single-run fabrication of a wide variety of nanowire-based optoelectronic devices.

  20. Does human-induced habitat transformation modify pollinator-mediated selection? A case study in Viola portalesia (Violaceae).

    PubMed

    Murúa, Maureen; Espinoza, Claudia; Bustamante, Ramiro; Marín, Víctor H; Medel, Rodrigo

    2010-05-01

    Pollinator-mediated selection is one of the most important factors driving adaptation in flowering plants. However, as ecological conditions change through habitat loss and fragmentation, the interactions among species may evolve in new and unexpected directions. Human-induced environmental variation is likely to affect selection regimes, but as yet no empirical examples have been reported. In the study reported here, we examined the influence of human-induced habitat transformation on the composition of pollinator assemblages and, hence, pollinator-mediated selection on the flower phenotype of Viola portalesia (Violaceae). Our results indicate that pollinator assemblages differed substantially in terms of species composition and visitation rate between nearby native and transformed habitats. Similarly, the insect species that contributed most to visitation rates differed between plant populations. While the magnitude and sign of pollinator-mediated selection on flower length and width did not differ between sites, selection for flower number lost significance in the transformed habitat, and a significant pattern of disruptive selection for flower shape, undetected in the native habitat, was present in the transformed one. Overall, the results of this study suggest that human-induced habitat change may not only modify the species composition of pollinator assemblages, relaxing the selection process on some flower characters, but they may also create new opportunities for fitness-trait covariation not present in pristine conditions.

  1. Agrobacterium-mediated transformation of the haploid liverwort Marchantia polymorpha L., an emerging model for plant biology.

    PubMed

    Ishizaki, Kimitsune; Chiyoda, Shota; Yamato, Katsuyuki T; Kohchi, Takayuki

    2008-07-01

    Agrobacterium-mediated transformation has not been practical in pteridophytes, bryophytes and algae to date, although it is commonly used in model plants including Arabidopsis and rice. Here we present a rapid Agrobacterium-mediated transformation system for the haploid liverwort Marchantia polymorpha L. using immature thalli developed from spores. Hundreds of hygromycin-resistant plants per sporangium were obtained by co-cultivation of immature thalli with Agrobacterium carrying the binary vector that contains a reporter, the beta-glucuronidase (GUS) gene with an intron, and a selection marker, the hygromycin phosphotransferase (hpt) gene. In this system, individual gemmae, which arise asexually from single initial cells, were analyzed as isogenic transformants. GUS activity staining showed that all hygromycin-resistant plants examined expressed the GUS transgene in planta. DNA analyses verified random integration of 1-5 copies of the intact T-DNA between the right and the left borders into the M. polymorpha genome. The efficient and rapid Agrobacterium-mediated transformation of M. polymorpha should provide molecular techniques to facilitate comparative genomics, taking advantage of this unique model plant that retains many features of the common ancestor of land plants.

  2. High Pressure Phase-Transformation Induced Texture Evolution and Strengthening in Zirconium Metal: Experiment and Modeling

    PubMed Central

    Yu, Xiaohui; Zhang, Ruifeng; Weldon, David; Vogel, Sven C.; Zhang, Jianzhong; Brown, Donald W.; Wang, Yanbin; Reiche, Helmut M.; Wang, Shanmin; Du, Shiyu; Jin, Changqing; Zhao, Yusheng

    2015-01-01

    We studied the phase-transition induced texture changes and strengthening mechanism for zirconium metal under quasi-hydrostatic compression and uni-axial deformation under confined high pressure using the deformation-DIA (D-DIA) apparatus. It is shown that the experimentally obtained texture for ω-phase Zr can be qualitatively described by combining a subset of orientation variants previously proposed in two different models. The determined flow stress for the high-pressure ω-phase is 0.5–1.2 GPa, more than three times higher than that of the α-phase. Using first-principles calculations, we investigated the mechanical and electronic properties of the two Zr polymorphs. We find that the observed strengthening can be attributed to the relatively strong directional bonding in the ω phase, which significantly increases its shear plastic resistance over the α-phase Zr. The present findings provide an alternate route for Zr metal strengthening by high-pressure phase transformation. PMID:26218405

  3. High pressure phase-transformation induced texture evolution and strengthening in zirconium metal: Experiment and modeling

    SciTech Connect

    Yu, Xiaohui; Zhang, Ruifeng; Weldon, David; Vogel, Sven C.; Zhang, Jianzhong; Brown, Donald W.; Wang, Yanbin; Reiche, Helmut M.; Wang, Shanmin; Du, Shiyu; Jin, Changqing; Zhao, Yusheng

    2015-07-28

    We studied the phase-transition induced texture changes and strengthening mechanism for zirconium metal under quasi-hydrostatic compression and uni-axial deformation under confined high pressure using the deformation-DIA (D-DIA) apparatus. It is shown that the experimentally obtained texture for ω-phase Zr can be qualitatively described by combining a subset of orientation variants previously proposed in two different models. The determined flow stress for the high-pressure ω-phase is 0.5–1.2 GPa, more than three times higher than that of the α-phase. Using first-principles calculations, we investigated the mechanical and electronic properties of the two Zr polymorphs. We find that the observed strengthening can be attributed to the relatively strong directional bonding in the ω phase, which significantly increases its shear plastic resistance over the α-phase Zr. The present findings provide an alternate route for Zr metal strengthening by high-pressure phase transformation.

  4. High pressure phase-transformation induced texture evolution and strengthening in zirconium metal: Experiment and modeling

    DOE PAGES

    Yu, Xiaohui; Zhang, Ruifeng; Weldon, David; ...

    2015-07-28

    We studied the phase-transition induced texture changes and strengthening mechanism for zirconium metal under quasi-hydrostatic compression and uni-axial deformation under confined high pressure using the deformation-DIA (D-DIA) apparatus. It is shown that the experimentally obtained texture for ω-phase Zr can be qualitatively described by combining a subset of orientation variants previously proposed in two different models. The determined flow stress for the high-pressure ω-phase is 0.5–1.2 GPa, more than three times higher than that of the α-phase. Using first-principles calculations, we investigated the mechanical and electronic properties of the two Zr polymorphs. We find that the observed strengthening can bemore » attributed to the relatively strong directional bonding in the ω phase, which significantly increases its shear plastic resistance over the α-phase Zr. The present findings provide an alternate route for Zr metal strengthening by high-pressure phase transformation.« less

  5. Mechanisms of Contact-Mediated Killing of Yeast Cells on Dry Metallic Copper Surfaces▿

    PubMed Central

    Quaranta, Davide; Krans, Travis; Santo, Christophe Espírito; Elowsky, Christian G.; Domaille, Dylan W.; Chang, Christopher J.; Grass, Gregor

    2011-01-01

    Surfaces made of copper or its alloys have strong antimicrobial properties against a wide variety of microorganisms. However, the molecular mode of action responsible for the antimicrobial efficacy of metallic copper is not known. Here, we show that dry copper surfaces inactivate Candida albicans and Saccharomyces cerevisiae within minutes in a process called contact-mediated killing. Cellular copper ion homeostasis systems influenced the kinetics of contact-mediated killing in both organisms. Deregulated copper ion uptake through a hyperactive S. cerevisiae Ctr1p (ScCtr1p) copper uptake transporter in Saccharomyces resulted in faster inactivation of mutant cells than of wild-type cells. Similarly, lack of the C. albicans Crp1p (CaCrp1p) copper-efflux P-type ATPase or the metallothionein CaCup1p caused more-rapid killing of Candida mutant cells than of wild-type cells. Candida and Saccharomyces took up large quantities of copper ions as soon as they were in contact with copper surfaces, as indicated by inductively coupled plasma mass spectroscopy (ICP-MS) analysis and by the intracellular copper ion-reporting dye coppersensor-1. Exposure to metallic copper did not cause lethality through genotoxicity, deleterious action on a cell's genetic material, as indicated by a mutation assay with Saccharomyces. Instead, toxicity mediated by metallic copper surfaces targeted membranes in both yeast species. With the use of Live/Dead staining, onset of rapid and extensive cytoplasmic membrane damage was observed in cells from copper surfaces. Fluorescence microscopy using the indicator dye DiSBaC2(3) indicated that cell membranes were depolarized. Also, during contact-mediated killing, vacuoles first became enlarged and then disappeared from the cells. Lastly, in metallic copper-stressed yeasts, oxidative stress in the cytoplasm and in mitochondria was elevated. PMID:21097600

  6. Seven-membered rings through metal-free rearrangement mediated by hypervalent iodine.

    PubMed

    Silva, Siguara Bastos Lemos; Torre, Adriana Della; de Carvalho, João Ernesto; Ruiz, Ana Lúcia Tasca Gois; Silva, Luiz F

    2015-01-15

    A versatile and metal-free approach for the synthesis of carbocycles and of heterocycles bearing seven- and eight-membered rings is described. The strategy is based on ring expansion of 1-vinylcycloalkanols (or the corresponding silyl or methyl ether) mediated by the hypervalent iodine reagent HTIB (PhI(OH)OTs). Reaction conditions can be easily adjusted to give ring expansion products bearing different functional groups. A route to medium-ring lactones was also developed.

  7. Critical role of retinoid/rexinoid signaling in mediating transformation and therapeutic response of NUP98-RARG leukemia.

    PubMed

    Qiu, J J; Zeisig, B B; Li, S; Liu, W; Chu, H; Song, Y; Giordano, A; Schwaller, J; Gronemeyer, H; Dong, S; So, C W E

    2015-05-01

    While the nucleoporin 98-retinoic acid receptor gamma (NUP98-RARG) is the first RARG fusion protein found in acute leukemia, its roles and the molecular basis in oncogenic transformation are currently unknown. Here, we showed that homodimeric NUP98-RARG not only acquired unique nuclear localization pattern and ability of recruiting both RXRA and wild-type NUP98, but also exhibited similar transcriptional properties as RARA fusions found in acute promyelocytic leukemia (APL). Using murine bone marrow retroviral transduction/transformation assay, we further demonstrated that NUP98-RARG fusion protein had gained transformation ability of primary hematopoietic stem/progenitor cells, which was critically dependent on the C-terminal GLFG domain of NUP98 and the DNA binding domain (DBD) of RARG. In contrast to other NUP98 fusions, cells transformed by the NUP98-RARG fusion were extremely sensitive to all-trans retinoic acid (ATRA) treatment. Interestingly, while pan-RXR agonists, SR11237 and LGD1069 could specifically inhibit NUP98-RARG transformed cells, mutation of the RXR interaction domain in NUP98-RARG had little effect on its transformation, revealing that therapeutic functions of rexinoid can be independent of the direct biochemical interaction between RXR and the fusion. Together, these results indicate that deregulation of the retinoid/rexinoid signaling pathway has a major role and may represent a potential therapeutic target for NUP98-RARG-mediated transformation.

  8. [Agrobacterium tumefaciens-mediated transformation of Aureobasidium pullulans and high-efficient screening for polymalic acid producing strain].

    PubMed

    Tu, Guangwei; Wang, Yongkang; Feng, Jun; Li, Xiaorong; Guo, Meijin; Zou, Xiang

    2015-07-01

    To develop a genetic transformation method of Aureobasidium pullulans and T-DNA insertion for high-efficient screening of polymalic acid (PMA) producing strain. Agrobacterium tumefaciens-AGL1, containing the selection genes encoding hygromycin B phosphotase or phosphinothricin acetyltranferase, was used to transform Aureobasidium pullulans CCTCC M2012223 and transformants were confirmed by colony PCR method. Transferred DNA (T-DNA) insertional mutants were cultured in microwell plate, and screened for high-titer PMA producing strain according to the pH response model. DNA walking was used to detect the insertion sites in the mutant. Results show that the selection markers could stably generated in the transformants, and 80 to 120 transformants could be found per 10(7) single cells. A high-titer PMA mutant H27 was obtained, giving a good PMA production caused by the disruption of phosphoglycerate mutase, that increased by 24.5% compared with the control. Agrobacterium tumefaciens-mediated transformation and high-efficient screening method were successfully developed, which will be helpful for genetic transformation of Aureobasidium pullulans and its functional genes discovery.

  9. High-efficiency Agrobacterium-mediated transformation of Norway spruce (Picea abies) and loblolly pine (Pinus taeda)

    NASA Technical Reports Server (NTRS)

    Wenck, A. R.; Quinn, M.; Whetten, R. W.; Pullman, G.; Sederoff, R.; Brown, C. S. (Principal Investigator)

    1999-01-01

    Agrobacterium-mediated gene transfer is the method of choice for many plant biotechnology laboratories; however, large-scale use of this organism in conifer transformation has been limited by difficult propagation of explant material, selection efficiencies and low transformation frequency. We have analyzed co-cultivation conditions and different disarmed strains of Agrobacterium to improve transformation. Additional copies of virulence genes were added to three common disarmed strains. These extra virulence genes included either a constitutively active virG or extra copies of virG and virB, both from pTiBo542. In experiments with Norway spruce, we increased transformation efficiencies 1000-fold from initial experiments where little or no transient expression was detected. Over 100 transformed lines expressing the marker gene beta-glucuronidase (GUS) were generated from rapidly dividing embryogenic suspension-cultured cells co-cultivated with Agrobacterium. GUS activity was used to monitor transient expression and to further test lines selected on kanamycin-containing medium. In loblolly pine, transient expression increased 10-fold utilizing modified Agrobacterium strains. Agrobacterium-mediated gene transfer is a useful technique for large-scale generation of transgenic Norway spruce and may prove useful for other conifer species.

  10. High-efficiency Agrobacterium-mediated transformation of Norway spruce (Picea abies) and loblolly pine (Pinus taeda)

    NASA Technical Reports Server (NTRS)

    Wenck, A. R.; Quinn, M.; Whetten, R. W.; Pullman, G.; Sederoff, R.; Brown, C. S. (Principal Investigator)

    1999-01-01

    Agrobacterium-mediated gene transfer is the method of choice for many plant biotechnology laboratories; however, large-scale use of this organism in conifer transformation has been limited by difficult propagation of explant material, selection efficiencies and low transformation frequency. We have analyzed co-cultivation conditions and different disarmed strains of Agrobacterium to improve transformation. Additional copies of virulence genes were added to three common disarmed strains. These extra virulence genes included either a constitutively active virG or extra copies of virG and virB, both from pTiBo542. In experiments with Norway spruce, we increased transformation efficiencies 1000-fold from initial experiments where little or no transient expression was detected. Over 100 transformed lines expressing the marker gene beta-glucuronidase (GUS) were generated from rapidly dividing embryogenic suspension-cultured cells co-cultivated with Agrobacterium. GUS activity was used to monitor transient expression and to further test lines selected on kanamycin-containing medium. In loblolly pine, transient expression increased 10-fold utilizing modified Agrobacterium strains. Agrobacterium-mediated gene transfer is a useful technique for large-scale generation of transgenic Norway spruce and may prove useful for other conifer species.

  11. A Type IV Pilus Mediates DNA Binding during Natural Transformation in Streptococcus pneumoniae

    PubMed Central

    Laurenceau, Raphaël; Péhau-Arnaudet, Gérard; Baconnais, Sonia; Gault, Joseph; Malosse, Christian; Dujeancourt, Annick; Campo, Nathalie; Chamot-Rooke, Julia; Le Cam, Eric; Claverys, Jean-Pierre; Fronzes, Rémi

    2013-01-01

    Natural genetic transformation is widely distributed in bacteria and generally occurs during a genetically programmed differentiated state called competence. This process promotes genome plasticity and adaptability in Gram-negative and Gram-positive bacteria. Transformation requires the binding and internalization of exogenous DNA, the mechanisms of which are unclear. Here, we report the discovery of a transformation pilus at the surface of competent Streptococcus pneumoniae cells. This Type IV-like pilus, which is primarily composed of the ComGC pilin, is required for transformation. We provide evidence that it directly binds DNA and propose that the transformation pilus is the primary DNA receptor on the bacterial cell during transformation in S. pneumoniae. Being a central component of the transformation apparatus, the transformation pilus enables S. pneumoniae, a major Gram-positive human pathogen, to acquire resistance to antibiotics and to escape vaccines through the binding and incorporation of new genetic material. PMID:23825953

  12. Transforming growth factor alpha is a critical mediator of radiation lung injury.

    PubMed

    Chung, Eun Joo; Hudak, Kathryn; Horton, Jason A; White, Ayla; Scroggins, Bradley T; Vaswani, Shiva; Citrin, Deborah

    2014-09-01

    Radiation fibrosis of the lung is a late toxicity of thoracic irradiation. Epidermal growth factor (EGF) signaling has previously been implicated in radiation lung injury. We hypothesized that TGF-α, an EGF receptor ligand, plays a key role in radiation-induced fibrosis in lung. Mice deficient in transforming growth factor (TGF-α(-/-)) and control C57Bl/6J (C57-WT) mice were exposed to thoracic irradiation in 5 daily fractions of 6 Gy. Cohorts of mice were followed for survival (n ≥ 5 per group) and tissue collection (n = 3 per strain and time point). Collagen accumulation in irradiated lungs was assessed by Masson's trichrome staining and analysis of hydroxyproline content. Cytokine levels in lung tissue were assessed with ELISA. The effects of TGF-α on pneumocyte and fibroblast proliferation and collagen production were analyzed in vitro. Lysyl oxidase (LOX) expression and activity were measured in vitro and in vivo. Irradiated C57-WT mice had a median survival of 24.4 weeks compared to 48.2 weeks for irradiated TGF-α(-/-) mice (P = 0.001). At 20 weeks after irradiation, hydroxyproline content was markedly increased in C57-WT mice exposed to radiation compared to TGF-α(-/-) mice exposed to radiation or unirradiated C57-WT mice (63.0, 30.5 and 37.6 μg/lung, respectively, P = 0.01). C57-WT mice exposed to radiation had dense foci of subpleural fibrosis at 20 weeks after exposure, whereas the lungs of irradiated TGF-α (-/-) mice were largely devoid of fibrotic foci. Lung tissue concentrations of IL-1β, IL-4, TNF-α, TGF-β and EGF at multiple time points after irradiation were similar in C57-WT and TGF-α(-/-) mice. TGF-α in lung tissue of C57-WT mice rose rapidly after irradiation and remained elevated through 20 weeks. TGF-α(-/-) mice had lower basal LOX expression than C57-WT mice. Both LOX expression and LOX activity were increased after irradiation in all mice but to a lesser degree in TGF-α(-/-) mice. Treatment of NIH-3T3 fibroblasts with TGF

  13. Transforming Growth Factor Alpha is a Critical Mediator of Radiation Lung Injury

    PubMed Central

    Chung, Eun Joo; Hudak, Kathryn; Horton, Jason A.; White, Ayla; Scroggins, Bradley T.; Vaswani, Shiva; Citrin, Deborah

    2014-01-01

    Radiation fibrosis of the lung is a late toxicity of thoracic irradiation. Epidermal growth factor (EGF) signaling has previously been implicated in radiation lung injury. We hypothesized that TGF-α, an EGF receptor ligand, plays a key role in radiation-induced fibrosis in lung. Mice deficient in transforming growth factor (TGF-α−/−) and control C57Bl/ 6J (C57-WT) mice were exposed to thoracic irradiation in 5 daily fractions of 6 Gy. Cohorts of mice were followed for survival (n ≥ 5 per group) and tissue collection (n = 3 per strain and time point). Collagen accumulation in irradiated lungs was assessed by Masson’s trichrome staining and analysis of hydroxyproline content. Cytokine levels in lung tissue were assessed with ELISA. The effects of TGF-α on pneumocyte and fibroblast proliferation and collagen production were analyzed in vitro. Lysyl oxidase (LOX) expression and activity were measured in vitro and in vivo. Irradiated C57-WT mice had a median survival of 24.4 weeks compared to 48.2 weeks for irradiated TGF-α−/− mice (P = 0.001). At 20 weeks after irradiation, hydroxyproline content was markedly increased in C57-WT mice exposed to radiation compared to TGF-α−/− mice exposed to radiation or unirradiated C57-WT mice (63.0, 30.5 and 37.6 µg/lung, respectively, P = 0.01). C57-WT mice exposed to radiation had dense foci of subpleural fibrosis at 20 weeks after exposure, whereas the lungs of irradiated TGF-α−/− mice were largely devoid of fibrotic foci. Lung tissue concentrations of IL-1β, IL-4, TNF-α, TGF-β and EGF at multiple time points after irradiation were similar in C57-WT and TGF-α−/− mice. TGF-α in lung tissue of C57-WT mice rose rapidly after irradiation and remained elevated through 20 weeks. TGF-α−/− mice had lower basal LOX expression than C57-WT mice. Both LOX expression and LOX activity were increased after irradiation in all mice but to a lesser degree in TGF-α−/− mice. Treatment of NIH-3T3

  14. Direct transformation of methyl imines to α-iminonitriles under mild and transition-metal-free conditions.

    PubMed

    Chen, Feng; Huang, Xiaoqiang; Cui, Yuxin; Jiao, Ning

    2013-08-19

    A novel transformation of methyl imines into α-iminonitriles under mild and transition-metal-free conditions is described. Three C sp 3-H bonds are cleaved in a radical pathway at room temperature under air. Simple bromide salts are employed to assist this radical process (see scheme; FG=functional group, PIDA = iodobenzene diacetate, TMS = trimethylsilyl).

  15. Microbial- and thiosulfate-mediated dissolution of mercury sulfide minerals and transformation to gaseous mercury

    DOE PAGES

    Vázquez-Rodríguez, Adiari I.; Hansel, Colleen M.; Zhang, Tong; ...

    2015-06-23

    Mercury (Hg) is a toxic heavy metal that poses significant environmental and human health risks. Soils and sediments, where Hg can exist as the Hg sulfide mineral metacinnabar (β-HgS), represent major Hg reservoirs in aquatic environments. Metacinnabar has historically been considered a sink for Hg in all but severely acidic environments, and thus disregarded as a potential source of Hg back to aqueous or gaseous pools. In this study, we conducted a combination of field and laboratory incubations to identify the potential for metacinnabar as a source of dissolved Hg within near neutral pH environments and the underpinning (a)biotic mechanismsmore » at play. We show that the abundant and widespread sulfur-oxidizing bacteria of the genus Thiobacillus extensively colonized metacinnabar chips incubated within aerobic, near neutral pH creek sediments. Laboratory incubations of axenic Thiobacillus thioparus cultures led to the release of metacinnabar-hosted Hg(II) and subsequent volatilization to Hg(0). This dissolution and volatilization was greatly enhanced in the presence of thiosulfate, which served a dual role by enhancing HgS dissolution through Hg complexation and providing an additional metabolic substrate for Thiobacillus. These findings reveal a new coupled abiotic-biotic pathway for the transformation of metacinnabar-bound Hg(II) to Hg(0), while expanding the sulfide substrates available for neutrophilic chemosynthetic bacteria to Hg-laden sulfides. Lastly, they also point to mineral-hosted Hg as an underappreciated source of gaseous elemental Hg to the environment.« less

  16. Microbial- and thiosulfate-mediated dissolution of mercury sulfide minerals and transformation to gaseous mercury

    SciTech Connect

    Vázquez-Rodríguez, Adiari I.; Hansel, Colleen M.; Zhang, Tong; Lamborg, Carl H.; Santelli, Cara M.; Webb, Samuel M.; Brooks, Scott C.

    2015-06-23

    Mercury (Hg) is a toxic heavy metal that poses significant environmental and human health risks. Soils and sediments, where Hg can exist as the Hg sulfide mineral metacinnabar (β-HgS), represent major Hg reservoirs in aquatic environments. Metacinnabar has historically been considered a sink for Hg in all but severely acidic environments, and thus disregarded as a potential source of Hg back to aqueous or gaseous pools. In this study, we conducted a combination of field and laboratory incubations to identify the potential for metacinnabar as a source of dissolved Hg within near neutral pH environments and the underpinning (a)biotic mechanisms at play. We show that the abundant and widespread sulfur-oxidizing bacteria of the genus Thiobacillus extensively colonized metacinnabar chips incubated within aerobic, near neutral pH creek sediments. Laboratory incubations of axenic Thiobacillus thioparus cultures led to the release of metacinnabar-hosted Hg(II) and subsequent volatilization to Hg(0). This dissolution and volatilization was greatly enhanced in the presence of thiosulfate, which served a dual role by enhancing HgS dissolution through Hg complexation and providing an additional metabolic substrate for Thiobacillus. These findings reveal a new coupled abiotic-biotic pathway for the transformation of metacinnabar-bound Hg(II) to Hg(0), while expanding the sulfide substrates available for neutrophilic chemosynthetic bacteria to Hg-laden sulfides. Lastly, they also point to mineral-hosted Hg as an underappreciated source of gaseous elemental Hg to the environment.

  17. A high-throughput Agrobacterium-mediated transformation system for the grass model species Brachypodium distachyon L.

    PubMed

    Păcurar, Daniel Ioan; Thordal-Christensen, Hans; Nielsen, Klaus Kristian; Lenk, Ingo

    2008-10-01

    In the ongoing process of developing Brachypodium distachyon as a model plant for temperate cereals and forage grasses, we have developed a high-throughput Agrobacterium-mediated transformation system for a diploid accession. Embryogenic callus, derived from immature embryos of the accession BDR018, were transformed with Agrobacterium tumefaciens strain AGL1 carrying two T-DNA plasmids, pDM805 and pWBV-Ds-Ubi-bar-Ds. Transient and stable transformation efficiencies were optimised by varying the pre-cultivation period, which had a strong effect on stable transformation efficiency. On average 55% of 17-day-old calli co-inoculated with Agrobacterium regenerated stable transgenic plants. Stable transformation frequencies of up to 80%, which to our knowledge is the highest transformation efficiency reported in graminaceous species, were observed. In a study of 177 transgenic lines transformed with pDM805, all of the regenerated transgenic lines were resistant to BASTA, while the gusA gene was expressed in 88% of the transgenic lines. Southern blot analysis revealed that 35% of the tested plants had a single T-DNA integration. Segregation analysis performed on progenies of ten selected T(0) plants indicated simple Mendelian inheritance of the two transgenes. Furthermore, the presence of two selection marker genes, bar and hpt, on the T-DNA of pWBV-Ds-Ubi-bar-Ds allowed us to characterize the developed transformation protocol with respect to full-length integration rate. Even when not selected for, full-length integration occurred in 97% of the transformants when using bialaphos as selection agent.

  18. Genetic transformation of Fusarium avenaceum by Agrobacterium tumefaciens mediated transformation and the development of a USER-Brick vector construction system

    PubMed Central

    2014-01-01

    Background The plant pathogenic and saprophytic fungus Fusarium avenaceum causes considerable in-field and post-field losses worldwide due to its infections of a wide range of different crops. Despite its significant impact on the profitability of agriculture production and a desire to characterize the infection process at the molecular biological level, no genetic transformation protocol has yet been established for F. avenaceum. In the current study, it is shown that F. avenaceum can be efficiently transformed by Agrobacterium tumefaciens mediated transformation. In addition, an efficient and versatile single step vector construction strategy relying on Uracil Specific Excision Reagent (USER) Fusion cloning, is developed. Results The new vector construction system, termed USER-Brick, is based on a limited number of PCR amplified vector fragments (core USER-Bricks) which are combined with PCR generated fragments from the gene of interest. The system was found to have an assembly efficiency of 97% with up to six DNA fragments, based on the construction of 55 vectors targeting different polyketide synthase (PKS) and PKS associated transcription factor encoding genes in F. avenaceum. Subsequently, the ΔFaPKS3 vector was used for optimizing A. tumefaciens mediated transformation (ATMT) of F. avenaceum with respect to six variables. Acetosyringone concentration, co-culturing time, co-culturing temperature and fungal inoculum were found to significantly impact the transformation frequency. Following optimization, an average of 140 transformants per 106 macroconidia was obtained in experiments aimed at introducing targeted genome modifications. Targeted deletion of FaPKS6 (FA08709.2) in F. avenaceum showed that this gene is essential for biosynthesis of the polyketide/nonribosomal compound fusaristatin A. Conclusion The new USER-Brick system is highly versatile by allowing for the reuse of a common set of building blocks to accommodate seven different types of genome

  19. Agrobacterium-Mediated Co-transformation of Multiple Genes in Metarhizium robertsii

    PubMed Central

    Bidochka, Michael J.

    2017-01-01

    Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii. PMID:28781540

  20. Possible stibnite transformation at the friction surface of the semi-metallic friction composites designed for car brake linings

    NASA Astrophysics Data System (ADS)

    Matějka, V.; Lu, Y.; Matějková, P.; Smetana, B.; Kukutschová, J.; Vaculík, M.; Tomášek, V.; Zlá, S.; Fan, Y.

    2011-12-01

    After a friction process several changes in phase composition of friction composites are often registered. High temperature, accompanied by high pressure induced during braking can cause initiation of chemical reactions which do not run at room or elevated temperatures under the atmospheric pressure. Most of the studies in the field of tribochemistry at friction surfaces of automotive semi-metallic brake linings deal with phenolic resin degradation and corrosion of metallic components. The paper addresses the formation of elemental antimony as well as the alloying process of iron with antimony observed on the surface of laboratory prepared semi-metallic friction composites containing stibnite. The role of alumina abrasives in the process of stibnite transformation is also discussed and mechanism of stibnite transformation was outlined.

  1. A stable and efficient Agrobacterium tumefaciens-mediated genetic transformation of the medicinal plant Digitalis purpurea L.

    PubMed

    Li, Ying; Gao, Zhenrui; Piao, Chunlan; Lu, Kaiwen; Wang, Zhiping; Cui, Min-Long

    2014-02-01

    In this study, we developed a rapid and efficient method for in vitro propagation and Agrobacterium tumefaciens-mediated transformation of Digitalis purpurea L. (syn. foxglove), an important medicinal plant. Mature leaf explants of D. purpurea were used for 100 % adventitious shoot regeneration on Murashige and Skoog (MS) medium supplemented with 1 mg L(-1) thidiazuron (TDZ) (a cytokine) and 0.1 mg L(-1) 1-naphthaleneacetic acid (NAA) (an auxin). Transformation was achieved by inoculating leaf explants with the A. tumefaciens strains GV2260/pBI121 or GV3101/pBI121. The binary vector pBI121 contained the reporter β-glucuronidase gene (GUS) and kanamycin selection marker nptII. Kanamycin-resistant shoots were regenerated directly on the selection medium 4-6 weeks after co-cultivation. Approximately, 52.2 and 60 % of kanamycin-resistant shoots transformed with Agrobacterium strains GV2260 and GV3101, respectively, showed strong GUS staining by histochemical assay. Furthermore, PCR and Southern blot analysis confirmed the presence of nptII and GUS on the chromosome of the transformed D. purpurea plants, and stable GUS expression was detected in the transformants by RT-PCR analysis. This efficient method of shoot regeneration and genetic transformation of D. purpurea will provide a powerful tool to increase and produce valuable components such as digitoxin, digoxin, and digoxigenin in D. purpurea through improved secondary metabolic pathways via a biotechnological approach.

  2. How Followers Differing in Career Motivation Gain Career Profits from Transformational Leaders: A Longitudinal Moderated Mediation Model

    PubMed Central

    Baethge, Anja; Rigotti, Thomas; Vincent-Hoeper, Sylvie

    2017-01-01

    Although, transformational leadership is among the most thoroughly examined leadership theories, knowledge regarding its association with followers' career outcomes is still limited. Furthermore, the underlying mechanisms explaining how transformational leaders affect their employees' career success are yet not well-understood. Based on theoretical assumptions about the processes involved in setting the goal of “making a career,” we propose an indirect effect of transformational leadership on subjective and objective career success via development opportunities that depends on the level of career motivation of employees. We conducted a longitudinal study with two measurement occasions separated by 13 months with 320 employees of a large IT company. Respondents provided ratings online on their direct supervisor's transformational leadership, their own development opportunities, and career motivation at T1; subjective career success was rated at both time points, whereas objective indicators of career transitions were rated at T2 retrospectively. Using structural equation modeling, we tested the proposed moderated mediation model. The results indicated that transformational leadership increased subordinates' subjective career success via development opportunities. In addition, and contrary to theoretical reasoning, the indirect effect was not significant for employees with high career motivation. Thus, employees high in career motivation appeared not to benefit from the development opportunities offered by transformational leaders. The results are discussed in light of tailored leadership that takes the aspirations, and needs of followers into account. PMID:28932204

  3. How Followers Differing in Career Motivation Gain Career Profits from Transformational Leaders: A Longitudinal Moderated Mediation Model.

    PubMed

    Baethge, Anja; Rigotti, Thomas; Vincent-Hoeper, Sylvie

    2017-01-01

    Although, transformational leadership is among the most thoroughly examined leadership theories, knowledge regarding its association with followers' career outcomes is still limited. Furthermore, the underlying mechanisms explaining how transformational leaders affect their employees' career success are yet not well-understood. Based on theoretical assumptions about the processes involved in setting the goal of "making a career," we propose an indirect effect of transformational leadership on subjective and objective career success via development opportunities that depends on the level of career motivation of employees. We conducted a longitudinal study with two measurement occasions separated by 13 months with 320 employees of a large IT company. Respondents provided ratings online on their direct supervisor's transformational leadership, their own development opportunities, and career motivation at T1; subjective career success was rated at both time points, whereas objective indicators of career transitions were rated at T2 retrospectively. Using structural equation modeling, we tested the proposed moderated mediation model. The results indicated that transformational leadership increased subordinates' subjective career success via development opportunities. In addition, and contrary to theoretical reasoning, the indirect effect was not significant for employees with high career motivation. Thus, employees high in career motivation appeared not to benefit from the development opportunities offered by transformational leaders. The results are discussed in light of tailored leadership that takes the aspirations, and needs of followers into account.

  4. Six1 overexpression at early stages of HPV16-mediated transformation of human keratinocytes promotes differentiation resistance and EMT

    SciTech Connect

    Xu, Hanwen; Pirisi, Lucia; Creek, Kim E.

    2015-01-01

    Previous studies in our laboratory discovered that SIX1 mRNA expression increased during in vitro progression of HPV16-immortalized human keratinocytes (HKc/HPV16) toward a differentiation-resistant (HKc/DR) phenotype. In this study, we explored the role of Six1 at early stages of HPV16-mediated transformation by overexpressing Six1 in HKc/HPV16. We found that Six1 overexpression in HKc/HPV16 increased cell proliferation and promoted cell migration and invasion by inducing epithelial–mesenchymal transition (EMT). Moreover, the overexpression of Six1 in HKc/HPV16 resulted in resistance to serum and calcium-induced differentiation, which is the hallmark of the HKc/DR phenotype. Activation of MAPK in HKc/HPV16 overexpressing Six1 is linked to resistance to calcium-induced differentiation. In conclusion, this study determined that Six1 overexpression resulted in differentiation resistance and promoted EMT at early stages of HPV16-mediated transformation of human keratinocytes. - Highlights: • Six1 expression increases during HPV16-mediated transformation. • Six1 overexpression causes differentiation resistance in HPV16-immortalized cells. • Six1 overexpression in HPV16-immortalized keratinocytes activates MAPK. • Activation of MAPK promotes EMT and differentiation resistance. • Six1 overexpression reduces Smad-dependent TGF-β signaling.

  5. The Role of the Secondary Coordination Sphere in Metal-Mediated Dioxygen Activation

    PubMed Central

    Shook, Ryan L.

    2012-01-01

    Alfred Werner proposed nearly 100 years ago that the secondary coordination sphere has a role in determining physical properties of transition metal complexes. We now know that the secondary coordination sphere impacts nearly all aspects of transition metal chemistry, including the reactivity and selectivity in metal-mediated processes. These features are highlighted in the binding and activation of dioxygen by transition metal complexes. There are clear connections between the control of the secondary coordination sphere and the ability of metal complexes to 1) reversibly bind dioxygen or 2) bind and activate dioxygen to form highly reactive M–oxo complexes. In this forum article, several biological and synthetic examples are presented and discussed in terms of structure-function relationships. Particular emphasis is given to systems with defined non-covalent interactions, such as intramolecular hydrogen bonds involving dioxygen-derived ligands. To further illustrate these effects, the homolytic cleavage of C–H bonds by M–oxo complexes with basic oxo ligands is described. PMID:20380466

  6. Factors influencing Agrobacterium-mediated embryogenic callus transformation of Valencia sweet orange (Citrus sinensis) containing the pTA29-barnase gene.

    PubMed

    Li, D D; Shi, W; Deng, X X

    2003-12-01

    Valencia sweet orange (Citrus sinensis (L.) Osbeck) calluses were used as explants to develop a new transformation system for citrus mediated by Agrobacterium tumefaciens. Factors affecting Agrobacterium-mediated transformation efficiency included mode of pre-cultivation, temperature of cocultivation and presence of acetosyringone (AS). The highest transformation efficiency was obtained with a 4-day pre-cultivation period in liquid medium. Transformation efficiency was higher when cocultivation was performed for 3 days at 19 degrees C than at 23 or 28 degrees C. Almost no resistant callus was obtained if the cocultivation medium lacked AS. The transformation procedure yielded transgenic Valencia plants containing the pTA29-barnase gene, as verified by PCR amplification and confirmed by Southern blotting. Because male sterility is a common factor leading to seedlessness in citrus cultivars with parthenocarpic characteristics, production of seedless citrus genotypes by Agrobacterium-mediated genetic transformation is a promising alternative to conventional breeding methods.

  7. Agrobacterium tumefaciens-mediated transformation of Lasiodiplodia theobromae, the causal agent of gummosis in cashew nut plants.

    PubMed

    Muniz, C R; da Silva, G F; Souza, M T; Freire, F C O; Kema, G H J; Guedes, M I F

    2014-02-21

    Lasiodiplodia theobromae is a major pathogen of many different crop cultures, including cashew nut plants. This paper describes an efficient Agrobacterium tumefaciens-mediated transformation (ATMT) system for the successful delivery of T-DNA, transferring the genes of green fluorescent protein (gfp) and hygromycin B phosphotransferase (hph) to L. theobromae. When the fungal pycnidiospores were co-cultured with A. tumefaciens harboring the binary vector with hph-gfp gene, hygromycin-resistant fungus only developed with acetosyringone supplementation. The cashew plants inoculated with the fungus expressing GFP revealed characteristic pathogen colonization by epifluorescence microscopy. Intense and bright green hyphae were observed for transformants in all extensions of mycelium cultures. The penetration of parenchyma cells near to the inoculation site, beneath the epicuticle surface, was observed prior to 25 dpi. Penetration was followed by the development of hyphae within invaded host cells. These findings provide a rapid and reproducible ATMT method for L. theobromae transformation.

  8. Agrobacterium tumefaciens-mediated transformation of Beauveria bassiana using an herbicide resistance gene as a selection marker.

    PubMed

    Fang, Weiguo; Zhang, Yongjun; Yang, Xingyong; Zheng, Xuelian; Duan, Hui; Li, Yi; Pei, Yan

    2004-01-01

    Beauveria bassiana has been investigated for use in the biological control of several insects in agricultural practice. To understand the molecular basis of virulence and host specificity and to improve the entomopathogenicity of B. bassiana, we have developed a simple, highly efficient and reliable Agrobacterium-mediated transformation method for B. bassiana using a phosphinothricin acetyltransferase (bar) gene as a selectable marker. Most transformants contained single copies of T-DNA and the T-DNA inserts were stably inherited after five generations. With this highly efficient transformation method for B. bassiana, we also obtained two putative T-DNA-tagged mutants that may have altered growth habits or virulence. Thus, the described protocol could provide a useful tool to manipulate the genetic make-up and to tag genes that may be important for virulence or growth and development of B. bassiana.

  9. Oxidative transformation of iron monosulfides and pyrite in estuarine sediments: Implications for trace metals mobilisation.

    PubMed

    Choppala, Girish; Bush, Richard; Moon, Ellen; Ward, Nicholas; Wang, Zhaohui; Bolan, Nanthi; Sullivan, Leigh

    2017-01-15

    Iron monosulfides are the initial iron sulfide minerals that form under reducing conditions in organic-rich sediments. Frequently referred as monosulfidic black ooze (MBO), these sediments exists in a range of anoxic systems including estuaries, coastal wetlands and permeable reactive barriers. The objective of this study was to investigate the transformation of solid phase sulfur, iron fractions and trace metals mobilisation in organic-rich hypersulfidic sediments during dredging. Two sediments from geographically contrasting sites in the Peel-Harvey Estuary were collected and subjected to oxidation through resuspension over 14 days. During oxidation, redox potential rapidly and continuously increased, although minimal change in pH was observed in both sediments. The majority of FeS was oxidised within 48 h. Although not as dynamic as FeS, unusually high rates of FeS2 oxidation were measured in both sediments at circumneutral pH, with between 39 and 58% of FeS2 oxidised over 14 days. The rapid oxidation of FeS2 may be attributed to the presence of nano-size FeS2 crystals (≈550-860 nm) with a high surface area. Before resuspension, solid bound Fe(total) was most abundant as measured by HCl-extractable Fe(II), followed by organic bound Fe(total) and oxide bound Fe(total). There was a marked decrease in these three fractions in both sediments during resuspension, with an increase in Fe(III) fraction. No significant release of trace metals was observed during resuspension of sulfidic sediments. However, disturbance to these estuarine sediments increases Fe(III) formation and further deteriorates the environment through smothering biological surfaces, deteriorating food sources and the quality of benthic habitats. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. AgarTrap-mediated genetic transformation using intact gemmae/gemmalings of the liverwort Marchantia polymorpha L.

    PubMed

    Tsuboyama-Tanaka, Shoko; Kodama, Yutaka

    2015-03-01

    The dioecious liverwort, Marchantia polymorpha L., is an emerging model plant. Various molecular biological techniques have been optimized for M. polymorpha for the past several years, and recently we reported a simplified Agrobacterium-mediated transformation method using sporelings (immature thalli from spores) of M. polymorpha. This method, termed AgarTrap (Agar-utilized Transformation with Pouring Solutions), completed by exchanging appropriate solutions on a single Petri dish to produce a sufficient number of independent transgenic sporelings. However, because spores are produced by crosses between males and females, the genetic backgrounds of resulting transgenic sporelings are not uniform. To easily produce transgenic liverworts with a uniform genetic background using AgarTrap, we developed an AgarTrap-mediated transformation method using intact gemmae/gemmalings produced by asexual reproduction. Using AgarTrap with male and female gemmae/gemmalings produced a sufficient number of independent transgenic gemmalings with uniform genetic backgrounds. The optimized transformation efficiencies were approximately 30 and 50 % in males and females, respectively. As with AgarTrap using sporelings, AgarTrap using intact gemmae/gemmalings will be useful in promoting studies of the molecular biology of M. polymorpha.

  11. Polydopamine-mediated surface-functionalization of graphene oxide for heavy metal ions removal

    SciTech Connect

    Dong, Zhihui; Zhang, Feng; Wang, Dong; Liu, Xia; Jin, Jian

    2015-04-15

    By utilizing polydopamine (PD) nano-thick interlayer as mediator, polyethylenimine (PEI) brushes with abundant amine groups were grafted onto the surface of PD coated graphene oxide (GO) uniformly via a Michael-Addition reaction and produced a PEI–PD/GO composite nanosheets. The PEI–PD/GO composite exhibited an improved performance for adsorption of heavy metal ions as compared to PEI-coated GO and pure GO. The adsorption capacities for Cu{sup 2+}, Cd{sup 2+}, Pb{sup 2+}, Hg{sup 2+} are up to 87, 106, 197, and 110 mg/g, respectively. To further make the GO based composite operable, PEI–PD/RGO aerogel was prepared through hydrothermal and achieved a high surface area up to 373 m{sup 2}/g. Although the adsorption capacity of PEI–PD/RGO aerogel for heavy metal ions decreases a little as compared to PEI–PD/GO composite dispersion (38, 32, 95, 113 mg/g corresponding to Cu{sup 2+}, Cd{sup 2+}, Pb{sup 2+}, and Hg{sup 2+}, respectively), it could be recycled several times in a simple way by releasing adsorbed metal ions, indicating its potential application for cleaning wastewater. - Graphical abstract: Polyethylenimine (PEI) brushes were grafted onto the surface of graphene oxide (GO) uniformly via a Michael-Addition reaction between the PEI and polydopamine interlayer coated on GO surface. The PEI–PD/GO composite exhibited an improved performance for adsorption of heavy metal ions compared to PEI-coated GO and pure GO. - Highlights: • We prepared polyethylenimine grafted polydopamine-mediated graphene oxide composites. • Introduction of PD layer increases metal ions adsorption capacity. • PEI–PD/RGO aerogel exhibited a superior adsorption performance. • PEI–PD/RGO aerogel can be recycled several times in a simple way.

  12. An efficient method for Agrobacterium-mediated genetic transformation and plant regeneration in cumin (Cuminum cyminum L.).

    PubMed

    Pandey, Sonika; Mishra, Avinash; Patel, Manish Kumar; Jha, Bhavanath

    2013-09-01

    Cumin is an annual herbaceous medicinally important plant having diverse applications. An efficient and reproducible method of Agrobacterium-mediated genetic transformation was herein established for the first time. A direct regeneration method without callus induction was optimised using embryos as explant material in Gamborg's B5 medium supplemented with 0.5-μM 6-benzyladenine and 2.0-μM α-naphthalene acetic acid. About 1,020 embryos (a mean of 255 embryos per batch) were used for the optimisation of transformation conditions. These conditions were an Agrobacterium cell suspension of 0.6 OD600, a co-cultivation time of 72 h, 300-μM acetosyringone and wounding of explants using a razor blade. Pre-cultured elongated embryos were treated using optimised conditions. About 720 embryos (a mean of 180 embryos per batch) were used for transformation and 95 % embryos showed transient β-glucuronidase expression after co-cultivation. Putative transformed embryos were cultured on B5 medium for shoot proliferation and 21 regenerated plants were obtained after selection and allowed to root. T0 plantlets showed β-glucuronidase expression and gene integration was confirmed via PCR amplification of 0.96 and 1.28 kb fragments of the hygromycin-phosphotransferase II and β-glucuronidase genes, respectively. In this study, a transformation efficiency of 1.5 % was demonstrated and a total of 11 transgenic plants were obtained at the hardening stage, however, only four plants acclimatised during hardening. Gene copy number was analysed by Southern blot analysis of hardened plants and single-copy gene integration was observed. This is the first successful attempt of Agrobacterium-mediated genetic transformation of cumin.

  13. Effects of sediment iron mineral composition on microbially mediated changes in divalent metal speciation: Importance of ferrihydrite

    NASA Astrophysics Data System (ADS)

    Cooper, D. Craig; Neal, Andrew L.; Kukkadapu, Ravi K.; Brewe, Dale; Coby, Aaron; Picardal, Flynn W.

    2005-04-01

    Dissimilatory metal reducing bacteria (DMRB) can influence geochemical processes that affect the speciation and mobility of metallic contaminants within natural environments. Most investigations into the effect of DMRB on sediment geochemistry utilize various synthetic oxides as the Fe III source (e.g., ferrihydrite, goethite, hematite). These synthetic materials do not represent the mineralogical composition of natural systems, and do not account for the effect of sediment mineral composition on microbially mediated processes. Our experiments with a DMRB ( Shewanella putrefaciens 200) and a divalent metal (Zn II) indicate that, while complexity in sediment mineral composition may not strongly impact the degree of "microbial iron reducibility," it does alter the geochemical consequences of such microbial activity. The ferrihydrite and clay mineral content are key factors. Microbial reduction of a synthetic blend of goethite and ferrihydrite (VHSA-G) carrying previously adsorbed Zn II increased both [Zn II-aq] and the proportion of adsorbed Zn II that is insoluble in 0.5 M HCl. Microbial reduction of Fe III in similarly treated iron-bearing clayey sediment (Fe-K-Q) and hematite sand, which contained minimal amounts of ferrihydrite, had no similar effect. Addition of ferrihydrite increased the effect of microbial Fe III reduction on Zn II association with a 0.5 M HCl insoluble phase in all sediment treatments, but the effect was inconsequential in the Fe-K-Q. Zinc k-edge X-ray absorption spectroscopy (XAS) data indicate that microbial Fe III reduction altered Zn II bonding in fundamentally different ways for VHSA-G and Fe-K-Q. In VHSA-G, ZnO 6 octahedra were present in both sterile and reduced samples; with a slightly increased average Zn-O coordination number and a slightly higher degree of long-range order in the reduced sample. This result may be consistent with enhanced Zn II substitution within goethite in the microbially reduced sample, though these data do not

  14. In situ-generated metal oxide catalyst during CO oxidation reaction transformed from redox-active metal-organic framework-supported palladium nanoparticles

    PubMed Central

    2012-01-01

    The preparation of redox-active metal-organic framework (ra-MOF)-supported Pd nanoparticles (NPs) via the redox couple-driven method is reported, which can yield unprotected metallic NPs at room temperature within 10 min without the use of reducing agents. The Pd@ra-MOF has been exploited as a precursor of an active catalyst for CO oxidation. Under the CO oxidation reaction condition, Pd@ra-MOF is transformed into a PdOx-NiOy/C nanocomposite to generate catalytically active species in situ, and the resultant nanocatalyst shows sustainable activity through synergistic stabilization. PMID:22898143

  15. Utilizing Hilbert-Huang transform in detection some of mechanical properties of the refractory metals

    NASA Astrophysics Data System (ADS)

    Mohammed, Arshed Abdulhamed; Haris, Sallehuddin Mohamed; Nuawi, Mohd Zaki

    2016-02-01

    This study is one of the first to report on the use of Hilbert-Huang transform (HHT) to determine the modulus of elasticity of a material, which is one of the most important properties of metals. In addition, this study involves an analytical study of the process of transfer of energy, which was represented in the form of intrinsic mode functions (IMFs). Moreover, the distribution of IMFs within the time-frequency-plain was determined by testing eight test specimens. Five test specimens were refractory materials, namely, Ti, Ti6AL4V, Zr, Nb, and Ta, and the other three were non-refractory materials, namely, Al, Brass, and ST4340. The new setup was composed of Mg and involves the use of two piezoelectric transducers, which were used as the emitter and receiver. The setup was designed and implemented in this research based on Mg usage to test the metals. First, a new relationship was derived between the pressure transmission coefficient (PTC) of the transmitted wave (through the emitter-water-test specimen-Mg to the receiver) and the corresponding values of the product of the density (ρ) and the modulus of elasticity (E) for the same test specimen. Another relationship was established between the PTCs and the total energy transmitted at high frequencies. This energy indicates the summation of IMFs that have high frequencies (THIMFs), higher than 10 kHz, can determine E better than TOF for most test specimens. To verify this results, with regard to the second conclusion, a new simulation for this setup was carried out using Simulink in MATLAB. Twelve theoretical tests were done, for high acoustic impedance metals, like Hf, Mo, WNiFe and W in addition to test the same group which was tested experimentally. The results of theoretical tests supported the experimental results except for Nb. Most of the conclusions were obtained through practical results and analytical studies. The results proved that THIMFs can determine the change in the microstructure of the alloys

  16. Plasmon‐Mediated Solar Energy Conversion via Photocatalysis in Noble Metal/Semiconductor Composites

    PubMed Central

    Wang, Mengye; Ye, Meidan; Iocozzia, James

    2016-01-01

    Plasmonics has remained a prominent and growing field over the past several decades. The coupling of various chemical and photo phenomenon has sparked considerable interest in plasmon‐mediated photocatalysis. Given plasmonic photocatalysis has only been developed for a relatively short period, considerable progress has been made in improving the absorption across the full solar spectrum and the efficiency of photo‐generated charge carrier separation. With recent advances in fundamental (i.e., mechanisms) and experimental studies (i.e., the influence of size, geometry, surrounding dielectric field, etc.) on plasmon‐mediated photocatalysis, the rational design and synthesis of metal/semiconductor hybrid nanostructure photocatalysts has been realized. This review seeks to highlight the recent impressive developments in plasmon‐mediated photocatalytic mechanisms (i.e., Schottky junction, direct electron transfer, enhanced local electric field, plasmon resonant energy transfer, and scattering and heating effects), summarize a set of factors (i.e., size, geometry, dielectric environment, loading amount and composition of plasmonic metal, and nanostructure and properties of semiconductors) that largely affect plasmonic photocatalysis, and finally conclude with a perspective on future directions within this rich field of research. PMID:27818901

  17. Plasmon-Mediated Solar Energy Conversion via Photocatalysis in Noble Metal/Semiconductor Composites.

    PubMed

    Wang, Mengye; Ye, Meidan; Iocozzia, James; Lin, Changjian; Lin, Zhiqun

    2016-06-01

    Plasmonics has remained a prominent and growing field over the past several decades. The coupling of various chemical and photo phenomenon has sparked considerable interest in plasmon-mediated photocatalysis. Given plasmonic photocatalysis has only been developed for a relatively short period, considerable progress has been made in improving the absorption across the full solar spectrum and the efficiency of photo-generated charge carrier separation. With recent advances in fundamental (i.e., mechanisms) and experimental studies (i.e., the influence of size, geometry, surrounding dielectric field, etc.) on plasmon-mediated photocatalysis, the rational design and synthesis of metal/semiconductor hybrid nanostructure photocatalysts has been realized. This review seeks to highlight the recent impressive developments in plasmon-mediated photocatalytic mechanisms (i.e., Schottky junction, direct electron transfer, enhanced local electric field, plasmon resonant energy transfer, and scattering and heating effects), summarize a set of factors (i.e., size, geometry, dielectric environment, loading amount and composition of plasmonic metal, and nanostructure and properties of semiconductors) that largely affect plasmonic photocatalysis, and finally conclude with a perspective on future directions within this rich field of research.

  18. Rare Earth Metal-Mediated Precision Polymerization of Vinylphosphonates and Conjugated Nitrogen-Containing Vinyl Monomers.

    PubMed

    Soller, Benedikt S; Salzinger, Stephan; Rieger, Bernhard

    2016-02-24

    This review focuses on introducing and explaining the rare earth metal-mediated group transfer polymerization (REM-GTP) of polar monomers and is composed of three main sections: poly(vinylphosphonate)s, surface-initiated group transfer polymerization (SI-GTP), and extension to N-coordinating Michael-type monomers (2-vinylpridine (2VP), 2-isopropenyl-2-oxazoline (IPOx)). The poly(vinylphosphonate)s section is divided into two parts: radical, anionic, and silyl ketene acetal group transfer polymerization (SKA-GTP) of vinylphosphonates in comparison to REM-GTP, and properties of poly(vinylphosphonate)s. The mechanism of vinylphosphonate REM-GTP is discussed in detail for initiation and propagation including activation enthalpies ΔH(‡) and entropies ΔS(‡) according to the Eyring equation. SI-GTP is presented as a method for surface functionalization, and recent trends for 2VP and IPOx polymerization are summarized. This review will serve as a good resource or guideline for researchers who are currently working in the field of rare earth metal mediated polymerization catalysis as well as for those who are interested in beginning to employ rare earth metal complexes for the synthesis of new materials from polar monomers.

  19. Ecological risk assessment on heavy metals in soils: Use of soil diffuse reflectance mid-infrared Fourier-transform spectroscopy

    PubMed Central

    Wang, Cheng; Li, Wei; Guo, Mingxing; Ji, Junfeng

    2017-01-01

    The bioavailability of heavy metals in soil is controlled by their concentrations and soil properties. Diffuse reflectance mid-infrared Fourier-transform spectroscopy (DRIFTS) is capable of detecting specific organic and inorganic bonds in metal complexes and minerals and therefore, has been employed to predict soil composition and heavy metal contents. The present study explored the potential of DRIFTS for estimating soil heavy metal bioavailability. Soil and corresponding wheat grain samples from the Yangtze River Delta region were analyzed by DRIFTS and chemical methods. Statistical regression analyses were conducted to correlate the soil spectral information to the concentrations of Cd, Cr, Cu, Zn, Pb, Ni, Hg and Fe in wheat grains. The principal components in the spectra influencing soil heavy metal bioavailability were identified and used in prediction model construction. The established soil DRIFTS-based prediction models were applied to estimate the heavy metal concentrations in wheat grains in the mid-Yangtze River Delta area. The predicted heavy metal concentrations of wheat grain were highly consistent with the measured levels by chemical analysis, showing a significant correlation (r2 > 0.72) with acceptable root mean square error RMSE. In conclusion, DRIFTS is a promising technique for assessing the bioavailability of soil heavy metals and related ecological risk. PMID:28198802

  20. Ecological risk assessment on heavy metals in soils: Use of soil diffuse reflectance mid-infrared Fourier-transform spectroscopy

    NASA Astrophysics Data System (ADS)

    Wang, Cheng; Li, Wei; Guo, Mingxing; Ji, Junfeng

    2017-02-01

    The bioavailability of heavy metals in soil is controlled by their concentrations and soil properties. Diffuse reflectance mid-infrared Fourier-transform spectroscopy (DRIFTS) is capable of detecting specific organic and inorganic bonds in metal complexes and minerals and therefore, has been employed to predict soil composition and heavy metal contents. The present study explored the potential of DRIFTS for estimating soil heavy metal bioavailability. Soil and corresponding wheat grain samples from the Yangtze River Delta region were analyzed by DRIFTS and chemical methods. Statistical regression analyses were conducted to correlate the soil spectral information to the concentrations of Cd, Cr, Cu, Zn, Pb, Ni, Hg and Fe in wheat grains. The principal components in the spectra influencing soil heavy metal bioavailability were identified and used in prediction model construction. The established soil DRIFTS-based prediction models were applied to estimate the heavy metal concentrations in wheat grains in the mid-Yangtze River Delta area. The predicted heavy metal concentrations of wheat grain were highly consistent with the measured levels by chemical analysis, showing a significant correlation (r2 > 0.72) with acceptable root mean square error RMSE. In conclusion, DRIFTS is a promising technique for assessing the bioavailability of soil heavy metals and related ecological risk.

  1. Highly Efficient Agrobacterium-Mediated Transformation of Wheat Via In Planta Inoculation

    NASA Astrophysics Data System (ADS)

    Risacher, Thierry; Craze, Melanie; Bowden, Sarah; Paul, Wyatt; Barsby, Tina

    This chapter details a reproducible method for the transformation of spring wheat using Agrobacterium tumefaciens via the direct inoculation of bacteria into immature seeds in planta as described in patent WO 00/63398(1. Transformation efficiencies from 1 to 30% have been obtained and average efficiencies of at least 5% are routinely achieved. Regenerated plants are phenotypically normal with 30-50% of transformation events carrying introduced genes at single insertion sites, a higher rate than is typically reported for transgenic plants produced using biolistic transformation methods.

  2. Agrobacterium rhizogenes-mediated transformation of Taraxacum platycarpum and changes of morphological characters.

    PubMed

    Lee, M H; Yoon, E S; Jeong, J H; Choi, Y E

    2004-06-01

    Transformed hairy roots were efficiently induced from seedlings of Taraxacum platycarpum by infection with Agrobacterium rhizogenes 15834. Root explants produced transformed roots at a higher frequency (76.5+/-3.5%) as compared to stem (32.7+/-4.8%) or cotyledon (16.2+/-5.7%). Hairy roots exhibited active elongation with high branching of roots on growth regulator-free medium. The competence of plant regeneration from non-transformed adventitious roots and transformed hairy roots was compared. The frequency of adventitious shoot formation from transformed roots was much higher (88.5+/-9.8%) than that of non-transformed roots (31.7 +/-9.5%) on hormone-free medium. Rooting of hairy root-derived adventitious shoots occurred easily on growth regulator-free medium but no rooting was observed on non-transformed shoots. The stable introduction of rol genes into Taraxacum plants was confirmed by PCR and Southern hybridization. Transgenic plantlets showed considerable differences in their morphology when compared to the corresponding wild-type (non-transgenic) plants. Plantlets formed from transformed roots had numerous fibrous roots with abundant lateral branches instead of the thickened taproots in non-transformed plants. The differences observed may reflect the modification of morphological root characters by introduction of rol genes.

  3. Water mediated alterations in gravity signal transform phytofilertation capability in hydroponic plants

    NASA Astrophysics Data System (ADS)

    Singh, Yogranjan; Singh Marabi, Rakesh; Satpute, Gyanesh Kumar; Mishra, Stuti

    2012-07-01

    An exorbitant sum of different synthetic molecules of chemicals including dyes and pigments are discharged into the environment, mainly via industrial effluents every year worldwide. The physical-chemical treatments for remediation viz adsorption, precipitation, ion exchange or filtration have proved to be disadvantageous because of high cost, low efficiency and inapplicability to a wide variety of dyes, or the formation of by-products and thereby creating waste disposal problems. Similarly the limited ability of micro-organisms to degrade xenobiotic especially sulphonoaromatic compounds, limits the efficiency and, therefore, the use of conventional wastewater treatment plants. In this context, the development of alternative biological treatments to eliminate these pollutants from industrial effluents is an important requirement. Plant metabolism, is extremely diverse and can be exploited to treat recalcitrant pollutants, not degradable by bacteria or fungi and can act as an important global sink for environmental pollutants. The presence of putative metabolites, in leaves of hydrophytes has been observed, indicating the transformation of several xenobiotics. A diverse range of the enzymes involved in the early stages of the detoxification process are closely associated with the redox biochemistry of the cell. The activities of enzymes such as glutathione transferases, peroxidases and cytochrome P450 monooxygenases and its multigenic family have implications with respect to the maintenance of redox homeostasis. Besides activating xenobiotics, cytochromes P450 is involved vitally in cell signaling for counteracting buoyant balance. Signal transduction cascades, including the role of cytochrome P450 monooxygenases in responding to gravitational cues, appear to be affected by buoyancy as well. Gravitropism is the orientation of growth in response to gravity and involves the perception of the gravitational force in the columella cells of the root cap where the primary

  4. A comparison of Agrobacterium-mediated transformation and protoplast-mediated transformation with CRISPR-Cas9 and bipartite gene targeting substrates, as effective gene targeting tools for Aspergillus carbonarius.

    PubMed

    Weyda, István; Yang, Lei; Vang, Jesper; Ahring, Birgitte K; Lübeck, Mette; Lübeck, Peter S

    2017-04-01

    In recent years, versatile genetic tools have been developed and applied to a number of filamentous fungi of industrial importance. However, the existing techniques have limitations when it comes to achieve the desired genetic modifications, especially for efficient gene targeting. In this study, we used Aspergillus carbonarius as a host strain due to its potential as a cell factory, and compared three gene targeting techniques by disrupting the ayg1 gene involved in the biosynthesis of conidial pigment in A. carbonarius. The absence of the ayg1 gene leads to phenotypic change in conidia color, which facilitated the analysis on the gene targeting frequency. The examined transformation techniques included Agrobacterium-mediated transformation (AMT) and protoplast-mediated transformation (PMT). Furthermore, the PMT for the disruption of the ayg1 gene was carried out with bipartite gene targeting fragments and the recently adapted CRISPR-Cas9 system. All three techniques were successful in generating Δayg1 mutants, but showed different efficiencies. The most efficient method for gene targeting was AMT, but further it was shown to be dependent on the choice of Agrobacterium strain. However, there are different advantages and disadvantages of all three gene targeting methods which are discussed, in order to facilitate future approaches for fungal strain improvements.

  5. NF-κB directly mediates epigenetic deregulation of common microRNAs in Epstein-Barr virus-mediated transformation of B-cells and in lymphomas

    PubMed Central

    Vento-Tormo, Roser; Rodríguez-Ubreva, Javier; Lisio, Lorena Di; Islam, Abul B. M. M. K.; Urquiza, Jose M.; Hernando, Henar; López-Bigas, Nuria; Shannon-Lowe, Claire; Martínez, Nerea; Montes-Moreno, Santiago; Piris, Miguel A.; Ballestar, Esteban

    2014-01-01

    MicroRNAs (miRNAs) have negative effects on gene expression and are major players in cell function in normal and pathological conditions. Epstein-Barr virus (EBV) infection of resting B lymphocytes results in their growth transformation and associates with different B cell lymphomas. EBV-mediated B cell transformation involves large changes in gene expression, including cellular miRNAs. We performed miRNA expression analysis in growth transformation of EBV-infected B cells. We observed predominant downregulation of miRNAs and upregulation of a few miRNAs. We observed similar profiles of miRNA expression in B cells stimulated with CD40L/IL-4, and those infected with EBNA-2- and LMP-1-deficient EBV particles, suggesting the implication of the NF-kB pathway, common to all four situations. In fact, the NF-kB subunit p65 associates with the transcription start site (TSS) of both upregulated and downregulated miRNAs following EBV infection This occurs together with changes at histone H3K27me3 and histone H3K4me3. Inhibition of the NF-kB pathway impairs changes in miRNA expression, NF-kB binding and changes at the above histone modifications near the TSS of these miRNA genes. Changes in expression of these miRNAs also occurred in diffuse large B cell lymphomas (DLBCL), which are strongly NF-kB dependent. Our results highlight the relevance of the NF-kB pathway in epigenetically mediated miRNA control in B cell transformation and DLBCL. PMID:25200074

  6. Mitochondrial Porin Isoform AtVDAC1 Regulates the Competence of Arabidopsis thaliana to Agrobacterium-Mediated Genetic Transformation

    PubMed Central

    Kwon, Tackmin

    2016-01-01

    The efficiency of Agrobacterium-mediated transformation in plants depends on the virulence of Agrobacterium strains, the plant tissue culture conditions, and the susceptibility of host plants. Understanding the molecular interactions between Agrobacterium and host plant cells is crucial when manipulating the susceptibility of recalcitrant crop plants and protecting orchard trees from crown gall disease. It was discovered that Arabidopsis voltage-dependent anion channel 1 (atvdac1) mutant has drastic effects on Agrobacterium-mediated tumorigenesis and growth developmental phenotypes, and that these effects are dependent on a Ws-0 genetic background. Genetic complementation of Arabidopsis vdac1 mutants and yeast porin1-deficient strain with members of the AtVDAC gene family revealed that AtVDAC1 is required for Agrobacterium-mediated transformation, and there is weak functional redundancy between AtVDAC1 and AtVDAC3, which is independent of porin activity. Furthermore, atvdac1 mutants were deficient in transient and stable transformation by Agrobacterium, suggesting that AtVDAC1 is involved in the early stages of Agrobacterium infection prior to transferred-DNA (T-DNA) integration. Transgenic plants overexpressing AtVDAC1 not only complemented the phenotypes of the atvdac1 mutant, but also showed high efficiency of transient T-DNA gene expression; however, the efficiency of stable transformation was not affected. Moreover, the effect of phytohormone treatment on competence to Agrobacterium was compromised in atvdac1 mutants. These data indicate that AtVDAC1 regulates the competence of Arabidopsis to Agrobacterium infection. PMID:27643450

  7. Involvement of HIF-2α-mediated inflammation in arsenite-induced transformation of human bronchial epithelial cells

    SciTech Connect

    Xu, Yuan; Zhao, Yue; Xu, Wenchao; Luo, Fei; Wang, Bairu; Li, Yuan; Pang, Ying; Liu, Qizhan

    2013-10-15

    Arsenic is a well established human carcinogen that causes diseases of the lung. Some studies have suggested a link between inflammation and lung cancer; however, it is unknown if arsenite-induced inflammation causally contributes to arsenite-caused malignant transformation of cells. In this study, we investigated the molecular mechanisms underlying inflammation during neoplastic transformation induced in human bronchial epithelial (HBE) cells by chronic exposure to arsenite. The results showed that, on acute or chronic exposure to arsenite, HBE cells over-expressed the pro-inflammatory cytokines, interleukin-6 (IL-6), interleukin-8 (IL-8), and interleukin-1β (IL-1β). The data also indicated that HIF-2α was involved in arsenite-induced inflammation. Moreover, IL-6 and IL-8 were essential for the malignant progression of arsenite-transformed HBE cells. Thus, these experiments show that HIF-2α mediates arsenite-induced inflammation and that such inflammation is involved in arsenite-induced malignant transformation of HBE cells. The results provide a link between the inflammatory response and the acquisition of a malignant transformed phenotype by cells chronically exposed to arsenite and thus establish a previously unknown mechanism for arsenite-induced carcinogenesis. - Highlights: • Arsenite induces inflammation. • Arsenite-induced the increases of IL-6 and IL-8 via HIF-2α. • Inflammation is involved in arsenite-induced carcinogenesis.

  8. Detection of Oil Palm Root Penetration by Agrobacterium-Mediated Transformed Ganoderma boninense, Expressing Green Fluorescent Protein.

    PubMed

    Govender, Nisha; Wong, Mui-Yun

    2017-04-01

    A highly efficient and reproducible Agrobacterium-mediated transformation protocol for Ganoderma boninense was developed to facilitate observation of the early stage infection of basal stem rot (BSR). The method was proven amenable to different explants (basidiospore, protoplast, and mycelium) of G. boninense. The transformation efficiency was highest (62%) under a treatment combination of protoplast explant and Agrobacterium strain LBA4404, with successful expression of an hyg marker gene and gus-gfp fusion gene under the control of heterologous p416 glyceraldehyde 3-phosphate dehydrogenase promoter. Optimal transformation conditions included a 1:100 Agrobacterium/explant ratio, induction of Agrobacterium virulence genes in the presence of 250 μm acetosyringone, co-cultivation at 22°C for 2 days on nitrocellulose membrane overlaid on an induction medium, and regeneration of transformants on potato glucose agar prepared with 0.6 M sucrose and 20 mM phosphate buffer. Evaluated transformants were able to infect root tissues of oil palm plantlets with needle-like microhyphae during the penetration event. The availability of this model pathogen system for BSR may lead to a better understanding of the pathogenicity factors associated with G. boninense penetration into oil palm roots.

  9. piggybac- and PhiC31-mediated genetic transformation of the Asian tiger mosquito, Aedes albopictus (Skuse).

    PubMed

    Labbé, Geneviève M C; Nimmo, Derric D; Alphey, Luke

    2010-08-17

    The Asian tiger mosquito, Aedes albopictus (Skuse), is a vector of several arboviruses including dengue and chikungunya. This highly invasive species originating from Southeast Asia has travelled the world in the last 30 years and is now established in Europe, North and South America, Africa, the Middle East and the Caribbean. In the absence of vaccine or antiviral drugs, efficient mosquito control strategies are crucial. Conventional control methods have so far failed to control Ae. albopictus adequately. Germline transformation of Aedes albopictus was achieved by micro-injection of embryos with a piggyBac-based transgene carrying a 3xP3-ECFP marker and an attP site, combined with piggyBac transposase mRNA and piggyBac helper plasmid. Five independent transgenic lines were established, corresponding to an estimated transformation efficiency of 2-3%. Three lines were re-injected with a second-phase plasmid carrying an attB site and a 3xP3-DsRed2 marker, combined with PhiC31 integrase mRNA. Successful site-specific integration was observed in all three lines with an estimated transformation efficiency of 2-6%. Both piggybac- and site-specific PhiC31-mediated germline transformation of Aedes albopictus were successfully achieved. This is the first report of Ae. albopictus germline transformation and engineering, a key step towards studying and controlling this species using novel molecular techniques and genetic control strategies.

  10. Improved Agrobacterium-mediated transformation and high efficiency of root formation from hypocotyl meristem of spring Brassica napus 'Precocity' cultivar.

    PubMed

    Liu, X X; Lang, S R; Su, L Q; Liu, X; Wang, X F

    2015-12-14

    Rape seed (Brassica napus L.) is one of the most important oil seed crops in the world. Genetic manipulation of rapeseed requires a suitable tissue culture system and an efficient method for plant regeneration, as well as an efficient transformation procedure. However, development of transgenic B. napus has been problematic, and current studies are limited to cultivated varieties. In this study, we report a protocol for regeneration of transgenic rape after Agrobacterium-mediated transformation of hypocotyls from the spring B. napus 'Precocity' cultivar. We analyzed the effects of plant growth regulators in the medium on regeneration. Additionally, factors affecting the transformation efficiency, including seedling age, Agrobacterium concentration, infection time, and co-cultivation time, were assessed by monitoring GUS expression. Results from these experiments revealed that transformation was optimized when the meristematic parts of the hypocotyls were taken from 8 day-old seedlings, cultured on Murashinge and Skoog basal media containing 0.1 mg/L 1-naphthaleneacetic acid and 2.5 mg/L 6-benzylaminopurine, and incubated in Agrobacterium suspension (OD600 = 0.5) for 3 to 5 min, followed by 2 days of co-cultivation. Integration of T-DNA into the plant genome was confirmed by polymerase chain reaction (PCR), b-glucuronidase histochemical staining, and quantitative real-time PCR. The protocols developed for regeneration, transformation, and rooting described in this study could help to accelerate the development of transgenic spring rape varieties with novel features.

  11. Advances in Photocatalysis: A Microreview of Visible Light Mediated Ruthenium and Iridium Catalyzed Organic Transformations.

    PubMed

    Day, Jon I; Teegardin, Kip; Weaver, Jimmie; Chan, John

    2016-07-15

    Photocatalytic organic transformations utilizing ruthenium and iridium complexes have garnered significant attention due to the access they provide to new synthetic spaces through new reaction mechanisms. A survey of the photophysical data and the diversity of transformations that may be accomplished utilizing commercially available photocatalysts is contained herein.

  12. Agrobacterium-mediated transformation of Fraxinus pennsylvanica hypocotyls and plant regeneration

    Treesearch

    Ningxia Du; Paula M. Pijut

    2009-01-01

    A genetic transformation protocol for green ash (Fraxinus pennsylvanica) hypocotyl explants was developed. Green ash hypocotyls were transformed using Agrobacterium tumefaciens strain EHA105 harboring binary vector pq35GR containing the neomycin phosphotransferase (nptII) and β-glucuronidase (GUS) fusion...

  13. Transformational Leadership and Creative Problem-Solving: The Mediating Role of Psychological Safety and Reflexivity

    ERIC Educational Resources Information Center

    Carmeli, Abraham; Sheaffer, Zachary; Binyamin, Galy; Reiter-Palmon, Roni; Shimoni, Tali

    2014-01-01

    Previous research has pointed to the importance of transformational leadership in facilitating employees' creative outcomes. However, the mechanism by which transformational leadership cultivates employees' creative problem-solving capacity is not well understood. Drawing on theories of leadership, information processing and creativity,…

  14. Transformational Leadership and Creative Problem-Solving: The Mediating Role of Psychological Safety and Reflexivity

    ERIC Educational Resources Information Center

    Carmeli, Abraham; Sheaffer, Zachary; Binyamin, Galy; Reiter-Palmon, Roni; Shimoni, Tali

    2014-01-01

    Previous research has pointed to the importance of transformational leadership in facilitating employees' creative outcomes. However, the mechanism by which transformational leadership cultivates employees' creative problem-solving capacity is not well understood. Drawing on theories of leadership, information processing and creativity,…

  15. Refined glufosinate selection in Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill].

    PubMed

    Zeng, P; Vadnais, D A; Zhang, Z; Polacco, J C

    2004-02-01

    Modern genetic analysis and manipulation of soybean ( Glycine max) depend heavily on an efficient and dependable transformation process, especially in public genotypes from which expressed sequence tag (EST), bacterial artificial chromosome and microarray data have been derived. Williams 82 is the subject of EST and functional genomics analyses. However, it has not previously been transformed successfully using either somatic embryogenesis-based or cotyledonary-node transformation methods, the two predominant soybean transformation systems. An advance has recently been made in using antioxidants to enhance Agrobacterium infection of soybean. Nonetheless, an undesirable effect of using these antioxidants is the compromised recovery of transgenic soybean when combined with the use of the herbicide glufosinate as a selective agent. Therefore, we optimized both Agrobacterium infection and glufosinate selection in the presence of L-cysteine for Williams 82. We have recovered transgenic lines of this genotype with an enhanced transformation efficiency using this herbicide selection system.

  16. Altering genomic integrity: heavy metal exposure promotes trans-posable element-mediated damage

    PubMed Central

    Morales, Maria E.; Servant, Geraldine; Ade, Catherine; Roy-Enge, Astrid M.

    2015-01-01

    Maintenance of genomic integrity is critical for cellular homeostasis and survival. The active transposable elements (TEs) composed primarily of three mobile element lineages LINE-1, Alu, and SVA comprise approximately 30% of the mass of the human genome. For the past two decades, studies have shown that TEs significantly contribute to genetic instability and that TE-caused damages are associated with genetic diseases and cancer. Different environmental exposures, including several heavy metals, influence how TEs interact with its host genome increasing their negative impact. This mini-review provides some basic knowledge on TEs, their contribution to disease and an overview of the current knowledge on how heavy metals influence TE-mediated damage. PMID:25774044

  17. Ammonia-Borane and Amine-Borane Dehydrogenation Mediated by Complex Metal Hydrides.

    PubMed

    Rossin, Andrea; Peruzzini, Maurizio

    2016-08-10

    This review is a comprehensive survey of the last 10 years of research on ammonia-borane and amine-borane dehydrogenation mediated by complex metal hydrides (CMHs), within the broader context of chemical hydrogen storage. The review also collects those cases where CMHs are the catalyst spent form or its resting state. Highlights on the reaction mechanism (strictly dependent on the CMH of choice) and the catalysts efficiency (in terms of equivalents of H2 produced and relative reaction rates) are provided throughout the discussion.

  18. Gold glyconanoparticles for mimics and measurement of metal ion-mediated carbohydrate-carbohydrate interactions.

    PubMed

    Reynolds, Angela J; Haines, Alan H; Russell, David A

    2006-01-31

    To mimic and measure calcium ion-mediated carbohydrate-carbohydrate interactions, four lactose derivatives have been synthesized for assembly on gold nanoparticles. The series of lactose derivatives varied by the length of the thiolated ethylene glycol anchor chain [O(CH2CH2O)(m)CH2CH2SH; where m = 0, 1, 2, and 3] used to self-assemble the carbohydrates to the preformed gold nanoparticles of ca. 16 nm diameter. Upon addition of calcium ions to the lactose-stabilized nanoparticles, rapid carbohydrate-carbohydrate interactions were visualized and subsequently measured using UV-visible spectrometry and transmission electron microscopy (TEM). The nanoparticle aggregates formed via metal-mediated carbohydrate-carbohydrate interactions could be readily redispersed through the addition of EDTA. Multiple reaggregation and redispersion cycles were achieved, confirming that the aggregation process was due to metal ion-mediated carbohydrate interactions rather than calcium chelation by residual citrate ions on the particle surface. The essential involvement of the lactose moiety in Ca2+ complexation was shown by control measurements on related D-glucose-derivatized nanoparticles, where a significantly reduced aggregation response was obtained only at high ion concentrations. Other group 2 metal ions with radii larger than that of calcium, viz., barium and strontium, were also shown to mediate the aggregation of the lactose-stabilized nanoparticles. The induced aggregation of the lactose nanoparticles was determined to be quantitatively dependent upon the calcium ion concentration. Furthermore, the analytical sensitivity of the calcium-induced aggregation and the linear dynamic range were dependent on the length of the ethylene glycol anchor chain. The shortest ethylene glycol chain (m = 0) gave the most sensitive response with the optimum limit of detection (0.8 mM Ca2+), whereas the longest ethylene glycol chain (m = 3) provides a measurement of calcium ion concentration

  19. Inhibition of CD1d-mediated antigen presentation by the transforming growth factor-β/Smad signalling pathway.

    PubMed

    Bailey, Jennifer C; Iyer, Abhirami K; Renukaradhya, Gourapura J; Lin, Yinling; Nguyen, Hoa; Brutkiewicz, Randy R

    2014-12-01

    CD1d-mediated lipid antigen presentation activates a subset of innate immune lymphocytes called invariant natural killer T (NKT) cells that, by virtue of their potent cytokine production, bridge the innate and adaptive immune systems. Transforming growth factor (TGF-β) is a known immune modulator that can activate the mitogen-activated protein kinase p38; we have previously shown that p38 is a negative regulator of CD1d-mediated antigen presentation. Several studies implicate a role for TGF-β in the activation of p38. Therefore, we hypothesized that TGF-β would impair antigen presentation by CD1d. Indeed, a dose-dependent decrease in CD1d-mediated antigen presentation and impairment of lipid antigen processing was observed in response to TGF-β treatment. However, it was found that this inhibition was not through p38 activation. Instead, Smads 2, 3 and 4, downstream elements of the TGF-β canonical signalling pathway, contributed to the observed effects. In marked contrast to that observed with CD1d, TGF-β was found to enhance MHC class II-mediated antigen presentation. Overall, these results suggest that the canonical TGF-β/Smad pathway negatively regulates an important arm of the host's innate immune responses - CD1d-mediated lipid antigen presentation to NKT cells.

  20. Transforming Environmental Knowledge into Behavior: The Mediating Role of Environmental Emotions

    ERIC Educational Resources Information Center

    Carmi, Nurit; Arnon, Sara; Orion, Nir

    2015-01-01

    The present study was based on the premise that environmental knowledge can drive environmental behavior only if it arouses environmental emotions. Using a structural equations modeling approach, we tested the direct, as well as the indirect (mediated) effects of knowledge on behavior and assessed the mediating role of environmental emotions. We…

  1. Transforming Environmental Knowledge into Behavior: The Mediating Role of Environmental Emotions

    ERIC Educational Resources Information Center

    Carmi, Nurit; Arnon, Sara; Orion, Nir

    2015-01-01

    The present study was based on the premise that environmental knowledge can drive environmental behavior only if it arouses environmental emotions. Using a structural equations modeling approach, we tested the direct, as well as the indirect (mediated) effects of knowledge on behavior and assessed the mediating role of environmental emotions. We…

  2. SacB-SacR gene cassette as the negative selection marker to suppress Agrobacterium overgrowth in Agrobacterium-mediated plant transformation

    USDA-ARS?s Scientific Manuscript database

    Agrobacterium overgrowth is a common problem in Agrobacterium-mediated plant transformation. To suppress the Agrobacterium overgrowth, various antibiotics have been used during plant tissue culture steps. The antibiotics are expensive and may adversely affect plant cell differentiation and reduce ...

  3. [Chlorination transformation and volatilization of heavy metals in fly ash from the incineration during the disposal process with higher temperature].

    PubMed

    Liu, Jing-Yong; Sun, Shui-Yu

    2012-09-01

    The volatilization of heavy metals Pb, Cd, Cu and Zn were studied at 900 degrees C and 1 000 degrees C with different residence time, meanwhile the influence of chlorination agents CaCl2, MgCl2, NaCl, FeCl3 and AlCl3 on heavy metals volatilization was studied. The results showed that the volatilization of heavy metals had great differences with the volatilization rate followed the order Pb > Cd > Zn > Cu, in which the volatilization rate of Pb was more than 80% and that of Cu was less than 30%. In the thermal disposal process, the volatilization rate influenced by temperature was greater than that by the residence time, and the volatile elements Pb and Cd were particularly evident in the volatilization. After adding chlorides in fly ash, the volatilization of heavy metals changed significantly, and the volatilization rate of the low volatile elements Cu and Zn increased significantly compared with Pb and Cd. With the content of chloride increasing, the volatilization rate of heavy metals increased, but different types of chloride compounds on heavy metal transformation were quite different, in which the promotion effect of NaCl on heavy metals Cd, Zn and Cu was less than that of other chlorinating agents. The results can provide strategies for the harmless disposal and maximize resource utilization and recycling of fly ash.

  4. Genetic transformation of Metroxylon sagu (Rottb.) cultures via Agrobacterium-mediated and particle bombardment.

    PubMed

    Ibrahim, Evra Raunie; Hossain, Md Anowar; Roslan, Hairul Azman

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.

  5. Genetic Transformation of Metroxylon sagu (Rottb.) Cultures via Agrobacterium-Mediated and Particle Bombardment

    PubMed Central

    Ibrahim, Evra Raunie

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance. PMID:25295258

  6. Agrobacterium tumefasciens-mediated transformation of the aquatic fungus Blastocladiella emersonii.

    PubMed

    Vieira, André L G; Camilo, César M

    2011-08-01

    Agrobacterium tumefaciens is widely used for plant DNA transformation and more recently, has also been used to transform yeast, filamentous fungi and even human cells. Using this technique, we developed the first transformation protocol for the saprobic aquatic fungus Blastocladiella emersonii, a Blastocladiomycete localized at the base of fungal phylogenetic tree, which has been shown as a promising and interesting model of study of cellular function and differentiation. We constructed binary T-DNA vectors containing hygromycin phosphotransferase (hph) or enhanced green fluorescent protein (egfp) genes, under the control of Aspergillus nidulans trpC promoter and terminator sequences. 24 h of co-cultivation in induction medium (IM) agar plates, followed by transfer to PYG-agar plates containing cefotaxim to kill Agrobacterium tumefsciens and hygromycin to select transformants, resulted in growth and sporulation of resistant transformants. Genomic DNA from the pool o resistant zoospores were shown to contain T-DNA insertion as evidenced by PCR amplification of hph gene. Using a similar protocol we could also evidence the expression of enhanced green fluorescent protein (EGFP) in zoospores derived from transformed cells. This protocol can also open new perspectives for other non-transformable closely related fungi, like the Chytridiomycete class.

  7. Parameters influencing Agrobacterium-mediated transformation system in safflower genotypes AKS-207 and PKV Pink.

    PubMed

    Dhumale, Dipti Raghunath; Shingote, Prashant Raghunath; Dudhare, Mahendra Shankarrao; Jadhav, Pravin Vishwanath; Kale, Prashant Bhaskar

    2016-12-01

    Shoot regeneration in safflower (Carthamus tinctorius 'AKS 207' and 'PKV Pink') genetically transformed using Agrobacterium was used for assessing various constraints to the efficiency of transformation including infection period, virulence induction medium, co-cultivation period, bacterial titre, selection regime, and the natural phenolic compound acetosyringone. Transformation frequency was promising with 8-10-day-old cotyledonary leaf explants. Therefore, explants of that age cultured on Agrobacterium minimal medium (AB) containing 100 µM acetosyringone were infected with Agrobacterium (cell titre 0.5 OD600nm) for 15 min followed by 48 h of co-cultivation on kanamycin-enriched medium (50 mg/L). Transformation of the shoots was confirmed using β-glucuronidase (GUS) histochemical assay and polymerase chain reaction (PCR). With the transformation protocol thus optimized, the transformation frequency as determined using GUS assays was 54.0 % for AKS 207 and 47.6 % for PKV Pink. The corresponding figures using PCR were 27.0 and 33.3 %. The transformed shoots required 10-14 weeks of culture initiation but produced very few roots.

  8. Agrobacterium tumefaciens-mediated transformation: An efficient tool for insertional mutagenesis and targeted gene disruption in Harpophora oryzae.

    PubMed

    Liu, Ning; Chen, Guo-Qing; Ning, Guo-Ao; Shi, Huan-Bin; Zhang, Chu-Long; Lu, Jian-Ping; Mao, Li-Juan; Feng, Xiao-Xiao; Liu, Xiao-Hong; Su, Zhen-Zhu; Lin, Fu-Cheng

    2016-01-01

    The endophytic filamentous fungus Harpophora oryzae is a beneficial endosymbiont isolated from the wild rice. H. oryzae could not only effectively improve growth rate and biomass yield of rice crops, but also induce systemic resistance against the rice blast fungus, Magnaporthe oryzae. In this study, Agrobacterium tumefaciens-mediated transformation (ATMT) was employed and optimized to modify the H. oryzae genes by either random DNA fragment integration or targeted gene replacement. Our results showed that co-cultivation of H. oryzae conidia with A. tumefaciens in the presence of acetosyringone for 48 h at 22 °C could lead to a relatively highest frequency of transformation, and 200 μM acetosyringone (AS) pre-cultivation of A. tumefaciens is also suggested. ATMT-mediated knockout mutagenesis was accomplished with the gene-deletion cassettes using a yeast homologous recombination method with a yeast-Escherichia-Agrobacterium shuttle vector pKOHo. Using the ATMT-mediated knockout mutagenesis, we successfully deleted three genes of H. oryzae (HoATG5, HoATG7, and HoATG8), and then got the null mutants ΔHoatg5, ΔHoatg7, and ΔHoatg8. These results suggest that ATMT is an efficient tool for gene modification including randomly insertional mutagenesis and gene deletion mutagenesis in H. oryzae. Copyright © 2015 Elsevier GmbH. All rights reserved.

  9. Aegle marmelos Mediated Green Synthesis of Different Nanostructured Metal Hexacyanoferrates: Activity against Photodegradation of Harmful Organic Dyes

    PubMed Central

    Jassal, Vidhisha; Kaith, B. S.

    2016-01-01

    Prussian blue analogue potassium metal hexacyanoferrate (KMHCF) nanoparticles Fe4[Fe(CN)6]3 (FeHCF), K2Cu3[Fe(CN)6]2 (KCuHCF), K2Ni[Fe(CN)6]·3H2O (KNiHCF), and K2Co[Fe(CN)6] (KCoHCF) have been synthesized using plant based biosurfactant Aegle marmelos (Bael) and water as a green solvent. It must be emphasized here that no harmful reagent or solvent was used throughout the study. Plant extracts are easily biodegradable and therefore do not cause any harm to the environment. Hence, the proposed method of synthesis of various KMHCF nanoparticles followed a green path. The synthesized nanoparticles were characterized by powder X-ray diffraction (PXRD), Field-Emission Scanning Electron Microscopy (FE-SEM), Transmission Electron Microscopy (TEM), and Fourier Transform Infrared Spectroscopy (FT-IR). MHCF nanoparticles were used for the photocatalytic degradation of toxic dyes like Malachite Green (MG), Eriochrome Black T (EBT), Methyl Orange (MO), and Methylene Blue (MB). Under optimized reaction conditions, maximum photocatalytic degradation was achieved in case of KCuHCF nanoparticles mediated degradation process (MG: 96.06%, EBT: 83.03%, MB: 94.72%, and MO: 63.71%) followed by KNiHCF (MG: 95%, EBT: 80.32%, MB: 91.35%, and MO: 59.42%), KCoHCF (MG: 91.45%, EBT: 78.84%, MB: 89.28%, and MO: 58.20%). PMID:27034896

  10. Porphyrin-Metalation-Mediated Tuning of Photoredox Catalytic Properties in Metal–Organic Frameworks

    SciTech Connect

    Johnson, Jacob A.; Luo, Jian; Zhang, Xu; Chen, Yu-Sheng; Morton, Martha D.; Echeverría, Elena; Torres, Fernand E.; Zhang, Jian

    2015-09-04

    Photoredox catalytic activation of organic molecules via single-electron transfer processes has proven to be a mild and efficient synthetic methodology. However, the heavy reliance on expensive ruthenium and iridium complexes limits their applications for scale-up synthesis. To this end, photoactive metal–organic frameworks (MOFs) exhibit unique advantages as novel heterogeneous photocatalytic systems, yet their utilization toward organic transformations has been limited. Here we describe the preparation and synthetic applications of four isostructural porphyrinic MOFs, namely, UNLPF-10a, -10b, -11, and -12, which are composed of free base, InIII-, SnIVCl2-, and SnIV-porphyrin building blocks, respectively. We demonstrate that the metalation with high valent metal cations (InIII and SnIV) significantly modifies the electronic structure of porphyrin macrocycle and provides a highly oxidative photoexcited state that can undergo efficient reductive quenching processes to facilitate organic reactions. In particular, UNLPF-12 exhibits both outstanding photostability and efficient photocatalytic activities toward a range of important organic transformations including aerobic hydroxylation of arylboronic acids, amine coupling, and the Mannich reaction.

  11. Transformation of Metal-Organic Frameworks/Coordination Polymers into Functional Nanostructured Materials: Experimental Approaches Based on Mechanistic Insights.

    PubMed

    Lee, Kyung Joo; Lee, Jae Hwa; Jeoung, Sungeun; Moon, Hoi Ri

    2017-10-09

    Nanostructured materials such as porous metal oxides, metal nanoparticles, porous carbons, and their composites have been intensively studied due to their applications, including energy conversion and storage devices, catalysis, and gas storage. Appropriate precursors and synthetic methods are chosen for synthesizing the target materials. About a decade ago, metal-organic frameworks (MOFs) and coordination polymers (CPs) emerged as new precursors for these nanomaterials because they contain both organic and inorganic species that can play parallel roles as both a template and a precursor under given circumstances. Thermal conversions of MOFs offer a promising toolbox for synthesizing functional nanomaterials that are difficult to obtain using conventional methods. Although understanding the conversion mechanism is important for designing MOF precursors for the synthesis of nanomaterials with desired physicochemical properties, comprehensive discussions revealing the transformation mechanism remain insufficient. This Account reviews the utilization of MOFs/CPs as precursors and their transformation into functional nanomaterials with a special emphasis on understanding the relationship between the intrinsic nature of the parent MOFs and the daughter nanomaterials while discussing various experimental approaches based on mechanistic insights. We discuss nanomaterials categorized by materials such as metal-based nanomaterials and porous carbons. For metal-based nanomaterials transformed from MOFs, the nature of metal ions in the MOF scaffolds affects the physicochemical properties of the resultant materials including the phase, composite, and morphology of nanomaterials. Organic ligands are also involved in the in situ chemical reactions with metal species during thermal conversion. We describe these conversion mechanisms by classifying the phase of metal components in the resultant materials. Along with the metal species, carbon is a major element in MOFs, and thus

  12. Metal-Organic Frameworks as Potential Platforms for Carbon Dioxide Capture and Chemical Transformation

    NASA Astrophysics Data System (ADS)

    Gao, Wenyang

    The anthropogenic carbon dioxide (CO2) emission into the atmosphere, mainly through the combustion of fossil fuels, has resulted in a balance disturbance of the carbon cycle. Overwhelming scientific evidence proves that the escalating level of atmospheric CO2 is deemed as the main culprit for global warming and climate change. It is thus imperative to develop viable CO2 capture and sequestration (CCS) technologies to reduce CO2 emissions, which is also essential to avoid the potential devastating effects in future. The drawbacks of energy-cost, corrosion and inefficiency for amine-based wet-scrubbing systems which are currently used in industry, have prompted the exploration of alternative approaches for CCS. Extensive efforts have been dedicated to the development of functional porous materials, such as activated carbons, zeolites, porous organic polymers, and metal-organic frameworks (MOFs) to capture CO2. However, these adsorbents are limited by either poor selectivity for CO2 separation from gas mixtures or low CO2 adsorption capacity. Therefore, it is still highly demanding to design next-generation adsorbent materials fulfilling the requirements of high CO2 selectivity and enough CO2 capacity, as well as high water/moisture stability under practical conditions. Metal-organic frameworks (MOFs) have been positioned at the forefront of this area as a promising type of candidate amongst various porous materials. This is triggered by the modularity and functionality of pore size, pore walls and inner surface of MOFs by use of crystal engineering approaches. In this work, several effective strategies, such as incorporating 1,2,3-triazole groups as moderate Lewis base centers into MOFs and employing flexible azamacrocycle-based ligands to build MOFs, demonstrate to be promising ways to enhance CO 2 uptake capacity and CO2 separation ability of porous MOFs. It is revealed through in-depth studies on counter-intuitive experimental observations that the local electric

  13. Compositional analysis of metal chelating materials using near-field photothermal Fourier transform infrared microspectroscopy.

    PubMed

    Moffat, Jonathan G; Mayes, Andrew G; Belton, Peter S; Craig, Duncan Q M; Reading, Mike

    2010-01-01

    Photothermal-Fourier transform-infrared (PT-FT-IR) microspectroscopy employs a thermal probe mounted in a scanning probe microscope (SPM). By placement of the tip of the probe on the surface of a solid sample, it can obtain localized IR spectra of a wide range of samples. A second mode of analysis is also available; a sample can be taken from the selected location using a technique called thermally assisted nanosampling (TAN), then a spectrum can be obtained of the nanosample while the probe is remote from the surface. We report a novel method of local compositional analysis that combines both of these types of measurement; a reagent is attached to the tip using TAN, then the reagent is placed in contact with analyte. IR spectroscopy can then be used to analyze any interaction between the reagent and surface it is placed in contact with. All of these modes of analysis were illustrated using a metal chelating agent. In the surface mode, changes to a solid bead of a chelating resin were measured using standard PT-FT-IR. In the nanosampling mode of analysis, a particle of a chelating polymer was attached to the tip of the probe using TAN and this was placed in contact with a concentrated calcium solution. Strong spectral changes were observed that mirrored those found when exposing the surface bound chelating resin bead to a solution of the same ion. A semiquantitative simulation of the PT spectrum for a chelating resin bead was achieved using a thermal diffusion model derived from photoacoustic spectroscopy indicating that semiquantitative or quantitative measurements will be possible in such a system.

  14. Functional Analysis and Discovery of Microbial Genes Transforming Metallic and Organic Pollutants: Database and Experimental Tools

    SciTech Connect

    Lawrence P. Wackett; Lynda B.M. Ellis

    2004-12-09

    Microbial functional genomics is faced with a burgeoning list of genes which are denoted as unknown or hypothetical for lack of any knowledge about their function. The majority of microbial genes encode enzymes. Enzymes are the catalysts of metabolism; catabolism, anabolism, stress responses, and many other cell functions. A major problem facing microbial functional genomics is proposed here to derive from the breadth of microbial metabolism, much of which remains undiscovered. The breadth of microbial metabolism has been surveyed by the PIs and represented according to reaction types on the University of Minnesota Biocatalysis/Biodegradation Database (UM-BBD): http://umbbd.ahc.umn.edu/search/FuncGrps.html The database depicts metabolism of 49 chemical functional groups, representing most of current knowledge. Twice that number of chemical groups are proposed here to be metabolized by microbes. Thus, at least 50% of the unique biochemical reactions catalyzed by microbes remain undiscovered. This further suggests that many unknown and hypothetical genes encode functions yet undiscovered. This gap will be partly filled by the current proposal. The UM-BBD will be greatly expanded as a resource for microbial functional genomics. Computational methods will be developed to predict microbial metabolism which is not yet discovered. Moreover, a concentrated effort to discover new microbial metabolism will be conducted. The research will focus on metabolism of direct interest to DOE, dealing with the transformation of metals, metalloids, organometallics and toxic organics. This is precisely the type of metabolism which has been characterized most poorly to date. Moreover, these studies will directly impact functional genomic analysis of DOE-relevant genomes.

  15. Agrobacterium-mediated transformation of safflower and the efficient recovery of transgenic plants via grafting

    PubMed Central

    2011-01-01

    Background Safflower (Carthamus tinctorius L.) is a difficult crop to genetically transform being susceptible to hyperhydration and poor in vitro root formation. In addition to traditional uses safflower has recently emerged as a broadacre platform for the production of transgenic products including modified oils and pharmaceutically active proteins. Despite commercial activities based on the genetic modification of safflower, there is no method available in the public domain describing the transformation of safflower that generates transformed T1 progeny. Results An efficient and reproducible protocol has been developed with a transformation efficiency of 4.8% and 3.1% for S-317 (high oleic acid content) and WT (high linoleic acid content) genotypes respectively. An improved safflower transformation T-DNA vector was developed, including a secreted GFP to allow non-destructive assessment of transgenic shoots. Hyperhydration and necrosis of Agrobacterium-infected cotyledons was effectively controlled by using iota-carrageenan, L-cysteine and ascorbic acid. To overcome poor in vitro root formation for the first time a grafting method was developed for safflower in which ~50% of transgenic shoots develop into mature plants bearing viable transgenic T1 seed. The integration and expression of secreted GFP and hygromycin genes were confirmed by PCR, Southern and Western blot analysis. Southern blot analysis in nine independent lines indicated that 1-7 transgenes were inserted per line and T1 progeny displayed Mendelian inheritance. Conclusions This protocol demonstrates significant improvements in both the efficiency and ease of use over existing safflower transformation protocols. This is the first complete method of genetic transformation of safflower that generates stably-transformed plants and progeny, allowing this crop to benefit from modern molecular applications. PMID:21595986

  16. Cadophora finlandia and Phialocephala fortinii: Agrobacterium-mediated transformation and functional GFP expression.

    PubMed

    Gorfer, Markus; Klaubauf, Sylvia; Bandian, Dragana; Strauss, Joseph

    2007-07-01

    Hygromycin B resistance was transferred to the sterile mycelia of Cadophora finlandia and Phialocephala fortinii by co-cultivation with Agrobacterium tumefaciens. Constitutively expressed green fluorescent protein (GFP) was also introduced using the same vector. Confocal laser scanning microscopy (CLSM) revealed strong fluorescence of transformants. Both traits were mitotically stable during one year of subculturing on non-selective growth medium. Southern blot analysis showed that the majority of the transformants contained single-copy integrations at random sites in the genome.

  17. Composition-Mediated Order-Disorder Transformation in FePt Nanoparticles

    SciTech Connect

    Johnston-Peck, Aaron C.; Cullen, David A.; Tracy, Joseph B.

    2013-07-08

    Thumbnail image of graphical abstract Heat-treated alloy FePt nanoparticles transform into L10 FePt and mixed L10/L12 FePt3 intermetallic phases. Enrichment in Pt in some nanoparticles, rather than intrinsic thermodynamic effects, drives phase segregation. FePt nanoparticles of uniform, equimolar composition are expected to transform into phase-pure, highly ordered L10 FePt nanoparticles.

  18. Susceptibility for N-ras-mediated transformation requires loss of tumor suppressor activity.

    PubMed

    Krizman, D B; Giovanella, B C; Tainsky, M A

    1990-01-01

    We have been using PA-1 human teratocarcinoma cells to study mechanisms by which oncogenes induce transformation. Tumorigenic PA-1 cells at passages greater than 100 (greater than P100) contain a spontaneously activated N-ras oncogene, while earlier-passage preneoplastic cells contain only the germ-line protooncogene and are nontumorigenic. One preneoplastic cell clone of PA-1 cells can be transformed by introduction of the cloned PA-1 N-ras in gene-transfer experiments, while another earlier-passage clonal cell line cannot be transformed. The goal of this investigation was to determine how human cells progress from resistance to susceptibility to ras oncogene-induced transformation. Somatic cell hybridization experiments described in this report indicate that the resistance of the low-passage cells to transformation is a dominant trait suppressing transformation. Loss of chromosomes from hybrid segregants suggested that tumor suppressors exist on chromosomes 1, 4, and 11. Extended in vitro passaging of somatic cell hybrids also resulted in the loss of chromosomes. Chromosome 1 was lost in these populations of cells, implying that reduction of this chromosome may promote proliferation and not specifically affect tumor formation.

  19. Establishment of a high efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.).

    PubMed

    Ozawa, Kenjirou

    2009-04-01

    Technologies for transformation of rice have been developed to meet the requirements of functional genomics in order to enable the production of transgenic rice plants with useful agricultural characters. However, many rice varieties are not efficiently transformed by Agrobacterium. We have succeeded in establishing a highly efficient transformation system in rice by co-cultivating rice calli with Agrobacterium on three filter papers moistened with enriched N6 or DKN media instead of using solid media. Rice calli immersed in Agrobacterium suspension (EHA101, Agrobacterium concentration of OD600=0.04) were co-cultured on three pieces of filter paper (9cm in diameter) moistened with 5.5mL of N6 or DKN liquid co-cultivation medium supplemented with 2,4-d (2mg/L), proline (10mM), casein hydrolysate (300mg/L), sucrose (30g/L), glucose (5g/L), l-cysteine (100mg/L) and acetosyringone (15mg/L) at 25°C for 3 days in the dark. Compared with the transformation efficiency of calli co-cultivated on solid media, transformation efficiency was increased by about fivefold by using the filter paper method for many varieties of rice, including those that previously yielded much poor transformation rates.

  20. Agrobacterium-mediated transformation of rough lemon (Citrus jambhiri Lush) with yeast HAL2 gene.

    PubMed

    Ali, Shawkat; Mannan, Abdul; El Oirdi, Mohamed; Waheed, Abdul; Mirza, Bushra

    2012-06-12

    Rough lemon (Citrus jambhiri Lush.) is the most commonly used Citrus rootstock in south Asia. It is extremely sensitive to salt stress that decreases the growth and yield of Citrus crops in many areas worldwide. Over expression of the yeast halotolerant gene (HAL2) results in increasing the level of salt tolerance in transgenic plants. Transformation of rough lemon was carried out by using Agrobacterium tumefaciens strains LBA4404 harboring plasmid pJRM17. Transgenic shoots were selected on kanamycin 100 mg L(-1) along with 250 mg L(-1) each of cefotaxime and vancomycin for effective inhibition of Agrobacterium growth. The Murashige and Skoog (MS) medium containing 200 μM acetoseryngone (AS) proved to be the best inoculation and co-cultivation medium for transformation. MS medium supplemented with 3 mg L(-1) of 6-benzylaminopurine (BA) showed maximum regeneration efficiency of the transformed explants. The final selection of the transformed plants was made on the basis of PCR and Southern blot analysis. Rough lemon has been successfully transformed via Agrobacterium tumefaciens with β-glucuronidase (GUS) and HAL2. Various factors affecting gene transformation and regeneration efficiency were also investigated.

  1. Vibration and glycerol-mediated plasmid DNA transformation for Escherichia coli.

    PubMed

    Shanehbandi, Dariush; Saei, Amir A; Zarredar, Habib; Barzegari, Abolfazl

    2013-11-01

    Escherichia coli transformation is an essential step in many molecular biology experiments. Despite earlier advances in the field, many studies including shotgun cloning still require more efficient transformation protocols. Chemical transformation has been the most popular method, in which competent cells are transformed following a brief period of heat shock. Here, we report a novel protocol with higher efficiency, in which competent E. coli cells (treated with CaCl2 ) grown in media containing glycerol experience a gentle vibration. Three E. coli strains DH5α, Jm107 and BL21 (DE3) and three plasmids pGEM-T, pET-28a and pCAMBIA with different sizes (3000, 5369 and 8428 bp, respectively) were used to test the protocol. The results indicated a significant increase in number of transformed colonies compared with heat-shock method. Our findings also demonstrated the favourable impacts of glycerol on transformation of E. coli.

  2. Agrobacterium-mediated transformation of rough lemon (Citrus jambhiri Lush) with yeast HAL2 gene

    PubMed Central

    2012-01-01

    Background Rough lemon (Citrus jambhiri Lush.) is the most commonly used Citrus rootstock in south Asia. It is extremely sensitive to salt stress that decreases the growth and yield of Citrus crops in many areas worldwide. Over expression of the yeast halotolerant gene (HAL2) results in increasing the level of salt tolerance in transgenic plants. Results Transformation of rough lemon was carried out by using Agrobacterium tumefaciens strains LBA4404 harboring plasmid pJRM17. Transgenic shoots were selected on kanamycin 100 mg L-1 along with 250 mg L-1 each of cefotaxime and vancomycin for effective inhibition of Agrobacterium growth. The Murashige and Skoog (MS) medium containing 200 μM acetoseryngone (AS) proved to be the best inoculation and co-cultivation medium for transformation. MS medium supplemented with 3 mg L-1 of 6-benzylaminopurine (BA) showed maximum regeneration efficiency of the transformed explants. The final selection of the transformed plants was made on the basis of PCR and Southern blot analysis. Conclusion Rough lemon has been successfully transformed via Agrobacterium tumefaciens with β-glucuronidase (GUS) and HAL2. Various factors affecting gene transformation and regeneration efficiency were also investigated. PMID:22691292

  3. 6-Pyrazolylpurine as an Artificial Nucleobase for Metal-Mediated Base Pairing in DNA Duplexes

    PubMed Central

    Léon, J. Christian; Sinha, Indranil; Müller, Jens

    2016-01-01

    The artificial nucleobase 6-pyrazol-1-yl-purine (6PP) has been investigated with respect to its usability in metal-mediated base pairing. As was shown by temperature-dependent UV spectroscopy, 6PP may form weakly stabilizing 6PP–Ag(I)–6PP homo base pairs. Interestingly, 6PP can be used to selectively recognize a complementary pyrimidine nucleobase. The addition of Ag(I) to a DNA duplex comprising a central 6PP:C mispair (C = cytosine) leads to a slight destabilization of the duplex. In contrast, a stabilizing 6PP–Ag(I)–T base pair is formed with a complementary thymine (T) residue. It is interesting to note that 6PP is capable of differentiating between the pyrimidine moieties despite the fact that it is not as sterically crowded as 6-(3,5-dimethylpyrazol-1-yl)purine, an artificial nucleobase that had previously been suggested for the recognition of nucleic acid sequences via the formation of a metal-mediated base pair. Hence, the additional methyl groups of 6-(3,5-dimethylpyrazol-1-yl)purine may not be required for the specific recognition of the complementary nucleobase. PMID:27089326

  4. The use of the phosphomannose isomerase gene as alternative selectable marker for Agrobacterium-mediated transformation of flax (Linum usitatissimum).

    PubMed

    Lamblin, Frédéric; Aimé, Aurélie; Hano, Christophe; Roussy, Isabelle; Domon, Jean-Marc; Van Droogenbroeck, Bart; Lainé, Eric

    2007-06-01

    In order to meet the future requirement of using non-antibiotic resistance genes for the production of transgenic plants, we have adapted the selectable marker system PMI/mannose to be used in Agrobacterium-mediated transformation of flax (Linum usitatissimum L.) cv. Barbara. The Escherichia coli pmi gene encodes a phosphomannose isomerase (E.C. 5.1.3.8) that converts mannose-6-phosphate, an inhibitor of glycolysis, into fructose-6-phosphate (glycolysis intermediate). Its expression in transformed cells allows them to grow on mannose-selective medium. The Agrobacterium tumefaciens strain GV3101 (pGV2260) harbouring the binary vector pNOV2819 that carries the pmi gene under the control of the Cestrum yellow leaf curling virus constitutive promoter was used for transformation experiments. Transgenic flax plants able to root on mannose-containing medium were obtained from hypocotyl-derived calli that had been selected on a combination of 20 g L(-1) sucrose and 10 g L(-1) mannose. Their transgenic state was confirmed by PCR and Southern blotting. Transgene expression was detected by RT-PCR in leaves, stems and roots of in vitro grown primary transformants. The mean transformation efficiency of 3.6%, that reached 6.4% in one experiment was comparable to that obtained when using the nptII selectable marker on the same cultivar. The ability of T1 seeds to germinate on mannose-containing medium confirmed the Mendelian inheritance of the pmi gene in the progeny of primary transformants. These results indicate that the PMI/mannose selection system can be successfully used for the recovery of flax transgenic plants under safe conditions for human health and the environment.

  5. Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14.

    PubMed

    Nyaboga, Evans N; Njiru, Joshua M; Tripathi, Leena

    2015-01-01

    Routine production of large numbers of transgenic plants is required to fully exploit advances in cassava biotechnology and support development of improved germplasm for deployment to farmers. This article describes an improved, high-efficiency transformation protocol for recalcitrant cassava cultivar TME14 preferred in Africa. Factors that favor production of friable embryogenic calli (FEC) were found to be use of DKW medium, crushing of organized embryogenic structures (OES) through 1-2 mm sized metal wire mesh, washing of crushed OES tissues and short exposure of tyrosine to somatic embryos; and transformation efficiency was enhanced by use of low Agrobacterium density during co-cultivation, co-centrifugation of FEC with Agrobacterium, germination of paramomycin resistant somatic embryos on medium containing BAP with gradual increase in concentration and variations of the frequency of subculture of cotyledonary-stage embryos on shoot elongation medium. By applying the optimized parameters, FEC were produced for cassava cultivar TME14 and transformed using Agrobacterium strain LBA4404 harboring the binary vector pCAMBIA2301. About 70-80 independent transgenic lines per ml settled cell volume (SCV) of FEC were regenerated on selective medium. Histochemical GUS assays confirmed the expression of gusA gene in transformed calli, somatic embryos and transgenic plants. The presence and integration of the gusA gene were confirmed by PCR and Southern blot analysis, respectively. RT-PCR analysis of transgenic plants confirmed the expression of gusA gene. This protocol demonstrates significantly enhanced transformation efficiency over existing cassava transformation protocols and could become a powerful tool for functional genomics and transferring new traits into cassava.

  6. Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14

    PubMed Central

    Nyaboga, Evans N.; Njiru, Joshua M.; Tripathi, Leena

    2015-01-01

    Routine production of large numbers of transgenic plants is required to fully exploit advances in cassava biotechnology and support development of improved germplasm for deployment to farmers. This article describes an improved, high-efficiency transformation protocol for recalcitrant cassava cultivar TME14 preferred in Africa. Factors that favor production of friable embryogenic calli (FEC) were found to be use of DKW medium, crushing of organized embryogenic structures (OES) through 1–2 mm sized metal wire mesh, washing of crushed OES tissues and short exposure of tyrosine to somatic embryos; and transformation efficiency was enhanced by use of low Agrobacterium density during co-cultivation, co-centrifugation of FEC with Agrobacterium, germination of paramomycin resistant somatic embryos on medium containing BAP with gradual increase in concentration and variations of the frequency of subculture of cotyledonary-stage embryos on shoot elongation medium. By applying the optimized parameters, FEC were produced for cassava cultivar TME14 and transformed using Agrobacterium strain LBA4404 harboring the binary vector pCAMBIA2301. About 70–80 independent transgenic lines per ml settled cell volume (SCV) of FEC were regenerated on selective medium. Histochemical GUS assays confirmed the expression of gusA gene in transformed calli, somatic embryos and transgenic plants. The presence and integration of the gusA gene were confirmed by PCR and Southern blot analysis, respectively. RT-PCR analysis of transgenic plants confirmed the expression of gusA gene. This protocol demonstrates significantly enhanced transformation efficiency over existing cassava transformation protocols and could become a powerful tool for functional genomics and transferring new traits into cassava. PMID:26113851

  7. The Ras-MAPK pathway downregulates Caveolin-1 in rodent fibroblast but not in human fibroblasts: implications in the resistance to oncogene-mediated transformation.

    PubMed

    Sasai, K; Kakumoto, K; Hanafusa, H; Akagi, T

    2007-01-18

    Normal human diploid fibroblasts (HDFs) are refractory to oncogene-mediated transformations in vitro, compared with rodent fibroblasts. As successful oncogene-mediated transformations of normal HDFs have been reported using the human telomerase catalytic subunit, it has been considered that telomerase activity contributes to the species-specific transformability. However, these transformed HDFs are much less malignant compared with those of rodent cells, suggesting the existence of undefined mechanisms that render HDFs resistant to malignant transformation. Here, cDNA microarray analysis identified caveolin-1 as one of the possible cellular factors involved in such mechanisms. The mitogen-activated protein kinases (MAPK) pathway downregulates Caveolin-1 in rodent fibroblasts, transformed by coexpression of the SV40 early region and activated H-Ras. In contrast, the coexpression of these two oncogenes in HDFs failed to reduce the expression level of Caveolin-1. These results strongly suggest the presence of critical differences in events following the phosphorylation of ERK during the activation process of the MAPK signaling pathway between human and rodent cells, as the ERK protein was similarly phosphorylated in both systems. Furthermore, the small interfering RNA-mediated suppression of Caveolin-1 facilitated the oncogene-mediated transformation of normal HDFs, clearly indicating that the differences in the transformability between human and rodent cells are due, at least in part, to the mechanism responsible for the resistance to Ras-induced Caveolin-1 downregulation in HDFs.

  8. The relationship between transformational teaching and adolescent physical activity: the mediating roles of personal and relational efficacy beliefs.

    PubMed

    Bourne, Jessica; Liu, Yan; Shields, Christopher A; Jackson, Ben; Zumbo, Bruno D; Beauchamp, Mark R

    2015-02-01

    The purpose of this study was to examine the extent to which transformational teaching, exhibited by secondary school physical education teachers, predicts within-class physical activity and leisure-time physical activity among adolescents. The study used a prospective observational design and involved data collected from 874 Grade 10 adolescents (M age = 15.41, (SD) = .61). Through use of structural equation modeling, the results revealed that adolescents' perceptions of transformational teaching were positively related to within-class physical activity and leisure-time physical activity, and these effects were mediated by adolescents' estimation of their teacher's confidence in their abilities (i.e. relation-inferred self-efficacy) and self-efficacy beliefs.

  9. Transgenic plants from shoot apical meristems of Vitis vinifera L. "Thompson Seedless" via Agrobacterium-mediated transformation.

    PubMed

    Dutt, M; Li, Z T; Dhekney, S A; Gray, D J

    2007-12-01

    Shoot apical meristem explants of Vitis vinifera "Thompson Seedless" were used for Agrobacterium-mediated genetic transformation. It was determined that the meristems had to be subjected to a dark growth phase then wounded to obtain transgenic plants. Morphological and histological studies illustrated the role of wounding to expose apical meristem cells for transformation. A bifunctional egfp/nptII fusion gene was used to select kanamycin resistant plants that expressed green fluorescent protein (GFP). Kanamycin at a concentration of 16 mg L(-1) in selection medium resulted in recovery of non-chimeric transgenic plants that uniformly expressed GFP, whereas 8 mg L(-1) kanamycin allowed non-transgenic and/or chimeric plants to develop. Polymerase chain reaction (PCR) and Southern blot analyses confirmed the presence of transgenes and their stable integration into the genome of regenerated plants. Up to 1% of shoot tips produced stable transgenic cultures within 6 weeks of treatment, resulting in a total of 18 independent lines.

  10. Formation of a new crystalline form of anhydrous β-maltose by ethanol-mediated crystal transformation.

    PubMed

    Verhoeven, Nicolas; Neoh, Tze Loon; Ohashi, Tetsuya; Furuta, Takeshi; Kurozumi, Sayaka; Yoshii, Hidefumi

    2012-04-01

    β-Maltose monohydrate was transformed into an anhydrous form by ethanol-mediated method under several temperatures with agitation. A new stable anhydrous form of β-maltose (Mβ(s)) was obtained, as substantiated by the X-ray diffraction patterns. Mβ(s) obtained by this method presented a fine porous structure, resulting in greater specific surface area compared to those of β-maltose monohydrate and anhydrous β-maltose obtained by vacuum drying (Mβ(h)). The crystal transformation presumably consisted of two steps: dehydration reaction from the hydrous to amorphous forms and crystal formation from the amorphous forms to the noble anhydrous form. The kinetics of these reactions were determined by thermal analysis using Jander's equation and Arrhenius plots. The overall activation energies of the dehydration reaction and the formation of anhydrous maltose were evaluated to be 100 and 90 kJ/mol, respectively.

  11. Temperature-mediated phase transformation, pore geometry and pore hysteresis transformation of borohydride derived in-born porous zirconium hydroxide nanopowders

    PubMed Central

    Nayak, Nadiya B.; Nayak, Bibhuti B.

    2016-01-01

    Development of in-born porous nature of zirconium hydroxide nanopowders through a facile hydrogen (H2) gas-bubbles assisted borohydride synthesis route using sodium borohydride (NaBH4) and novel information on the temperature-mediated phase transformation, pore geometry as well as pore hysteresis transformation of in-born porous zirconium hydroxide nanopowders with the help of X-ray diffraction (XRD), Brunauer–Emmett–Teller (BET) isotherm and Transmission Electron Microscopy (TEM) images are the main theme of this research work. Without any surfactants or pore forming agents, the borohydride derived amorphous nature of porous powders was stable up to 500 °C and then the seed crystals start to develop within the loose amorphous matrix and trapping the inter-particulate voids, which led to develop the porous nature of tetragonal zirconium oxide at 600 °C and further sustain this porous nature as well as tetragonal phase of zirconium oxide up to 800 °C. The novel hydrogen (H2) gas-bubbles assisted borohydride synthesis route led to develop thermally stable porous zirconium hydroxide/oxide nanopowders with an adequate pore size, pore volume, and surface area and thus these porous materials are further suggested for promising use in different areas of applications. PMID:27198738

  12. Development of a phosphomannose isomerase-based Agrobacterium-mediated transformation system for chickpea (Cicer arietinum L.).

    PubMed

    Patil, Gunvant; Deokar, Amit; Jain, P K; Thengane, R J; Srinivasan, R

    2009-11-01

    To develop an alternative genetic transformation system that is not dependent on an antibiotic selection strategy, the phosphomannose isomerase gene (pmi) system was evaluated for producing transgenic plants of chickpea (Cicer arietinum L.). A shoot morphogenesis protocol based on the thidiazuron (TDZ)-induced shoot morphogenesis system was combined with Agrobacterium-mediated transformation of the pmi gene and selection of transgenic plants on mannose. Embryo axis explants of chickpea cv. C-235 were grown on a TDZ-supplemented medium for shoot proliferation. Embryo axis explants from which the first and second flush of shoots were removed were transformed using Agrobacterium carrying the pmi gene, and emerging shoots were allowed to regenerate on a zeatin-supplemented medium with an initial selection pressure of 20 g l(-1) mannose. Rooting was induced in the selected shoots on an indole-3-butyric acid (IBA)-supplemented medium with a selection pressure of 15 g l(-1) mannose. PCR with marker gene-specific primers and chlorophenol red (CPR) assay of the shoots indicated that shoots had been transformed. RT-PCR and Southern analysis of selected regenerated plants further confirmed integration of the transgene into the chickpea genome. These positive results suggest that the pmi/mannose selection system can be used to produce transgenic plants of chickpea that are free from antibiotic resistance marker genes.

  13. Improving the sensitivity of a high-Tc SQUID at MHz frequency using a normal metal transformer

    NASA Astrophysics Data System (ADS)

    He, D. F.; Itozaki, H.; Tachiki, M.

    2006-05-01

    Superconducting quantum interference devices (SQUIDs) can be used to detect the signals of nuclear quadrupole resonance (NQR). The NQR frequencies of some interesting materials are in the order of MHz. However, the sensitivity of a high-Tc SQUID is normally not enough to detect the weak NQR signals. To improve the sensitivity of a high-Tc SQUID at MHz frequency, we used a transformer made of normal copper wire. The transformer was composed of a pickup coil, an input coil and a capacitor. The pickup coil was used to detect the magnetic field; the input coil was used to couple the field to the SQUID; and the capacitor was used to create a resonant frequency. By using the normal metal transformer, the magnetic field resolution of the high-Tc dc SQUID was improved by about 38.8 times (from 220 to 5.67 fT Hz-1/2) at 954 kHz.

  14. Transgenic grasspea (Lathyrus sativus L.): factors influencing agrobacterium-mediated transformation and regeneration.

    PubMed

    Barik, D P; Mohapatra, U; Chand, P K

    2005-11-01

    A reproducible procedure was developed for genetic transformation of grasspea using epicotyl segment co-cultivation with Agrobacterium. Two disarmed Agrobacterium tumefaciens strains, EHA 105 and LBA 4404, both carrying the binary plasmid p35SGUSINT with the neomycin phosphotransferase II (nptII) gene and the beta-glucuronidase (gus)-intron, were studied as vector systems. The latter was found to have a higher transforming ability. Several key factors modifying the transformation rate were optimized. The highest transformation rate was achieved using hand-pricked explants for infection with an Agrobacterium culture corresponding to OD(600) congruent with 0.6 and diluted to a cell density of 10(9) cells ml(-1) for 10 min, followed by co-cultivation for 4 days in a medium maintained at pH 5.6. Putative transformed explants capable of forming shoots were selected on regeneration medium containing kanamycin (100 mug ml(-1)). We achieved up to 36% transient expression based on the GUS histochemical assay. Southern hybridization of genomic DNA of the kanamycin-resistant GUS-expressive shoots to a gus-intron probe substantiated the integration of the transgene. Transformed shoots were rooted on half-strength MS containing 0.5 mg l(-1) indole-3-acetic acid, acclimated in vermi-compost and established in the experimental field. Germ-line transformation was evident through progeny analysis. Among T(1) seedlings of most transgenic plant lines, kanamycin-resistant and -sensitive plants segregated in a ratio close to 3:1.

  15. Enhancers of Agrobacterium-mediated Transformation of Tibouchina semidecandra Selected on the Basis of GFP Expression.

    PubMed

    Yong, Wilson Thau Lym; Henry, Erle Stanley; Abdullah, Janna Ong

    2010-12-01

    Genetic engineering is a powerful tool for the improvement of plant traits. Despite reported successes in the plant kingdom, this technology has barely scratched the surface of the Melastomataceae family. Limited studies have led to some optimisation of parameters known to affect the transformation efficiency of these plants. The major finding of this study was to optimise the presence of selected enhancers [e.g., monosaccharides (D-glucose, D-galactose and D-fructose), tyrosine, aluminium chloride (AICI3) and ascorbic acid] to improve the transformation efficiency of Tibouchina semidecandra. Agrobacterium tumefaciens strain LBA4404 harbouring the disarmed plasmid pCAMBIA1304 was used to transform shoots and nodes of T. semidecandra. Different concentrations of the transformation enhancers were tested by using green fluorescent protein (GFP) as a reporter. The results obtained were based on the percentage of GFP expression, which was observed 14 days post-transformation. A combination of 120 μM galactose and 100 μM tyrosine supplemented with 600 μM AICI3 in the presence of 15 mg/l ascorbic acid gave the highest percentage of positive transformants for T. semidecandra shoots. Whereas 60 μM galactose and 50 μM tyrosine with 200 μM AICI3 in the presence of 15 mg/l ascorbic acid was optimum for T. semidecandra nodes. The presence of the hygromycin phosphotransferase II (hptII) transgene in the genomic DNA of putative T. semidecandra transformants was verified by PCR amplification with specific primers.

  16. Gene regulation mediating fiber-type transformation in skeletal muscle cells is partly glucose- and ChREBP-dependent.

    PubMed

    Hanke, Nina; Scheibe, Renate J; Manukjan, Georgi; Ewers, David; Umeda, Patrick K; Chang, Kin-Chow; Kubis, Hans-Peter; Gros, Gerolf; Meissner, Joachim D

    2011-03-01

    Adaptations in the oxidative capacity of skeletal muscle cells can occur under several physiological or pathological conditions. We investigated the effect of increasing extracellular glucose concentration on the expression of markers of energy metabolism in primary skeletal muscle cells and the C2C12 muscle cell line. Growth of myotubes in 25mM glucose (high glucose, HG) compared with 5.55mM led to increases in the expression and activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a marker of glycolytic energy metabolism, while oxidative markers peroxisome proliferator-activated receptor γ coactivator 1α and citrate synthase decreased. HG induced metabolic adaptations as are seen during a slow-to-fast fiber transformation. Furthermore, HG increased fast myosin heavy chain (MHC) IId/x but did not change slow MHCI/β expression. Protein phosphatase 2A (PP2A) was shown to mediate the effects of HG on GAPDH and MHCIId/x. Carbohydrate response element-binding protein (ChREBP), a glucose-dependent transcription factor downstream of PP2A, partially mediated the effects of glucose on metabolic markers. The glucose-induced increase in PP2A activity was associated with an increase in p38 mitogen-activated protein kinase activity, which presumably mediates the increase in MHCIId/x promoter activity. Liver X receptor, another possible mediator of glucose effects, induced only an incomplete metabolic shift, mainly increasing the expression of the glycolytic marker. Taken together, HG induces a partial slow-to-fast transformation comprising metabolic enzymes together with an increased expression of MHCIId/x. This work demonstrates a functional role for ChREBP in determining the metabolic type of muscle fibers and highlights the importance of glucose as a signaling molecule in muscle. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. [The effect of solid phase transformation on the metal-ceramic compatibility of Co-Cr alloy].

    PubMed

    Wu, Zhikai; Xu, Sheng; Li, Ning

    2011-12-01

    To study the effect of solid phase transformation on the metal-ceramic compatibility of Co-Cr alloy during firing programs. 9 foils of Co-Cr and Ni-Cr alloy with the dimension of 25 mmx3 mmx0.5 mm were casted using lost wax technology respectively. Among them, 6 specimens were subjected to metal-ceramic bonding strength test (three point bending method), the ceramic layer of 3 specimens were removed for X-ray diffraction (XRD) analysis. One cylindrical specimen of each alloy was casted for the test of coefficient of thermal expansion, cooling curves were recorded. The metal-ceramic bonding strength of Ni-Cr, Co-Cr alloy was (49.1 +/- 3.40), (40.9 +/- 2.02) MPa respectively. There was significant difference between the two groups' bonding strength (P = 0.00). The coefficient of thermal expansion in the 20-500 degrees C interval of Ni-Cr and Co-Cr alloy was 13.9 x 10(-6), 13.8 x 10(-6) x K(-1) respectively. XRD analysis indicated that the microstructure of Ni-Cr alloy was austenite. While Co-Cr alloy was constituted by fcc phase, hcp phase and sigma phase. During the cooling procedure, the transformation of fcc phase to hcp phase and segregation of needle-like sigma phase intensify the linear contraction speed of Co-Cr alloy, which decreases the metal-ceramic adaptability.

  18. [Analysis of heavy-metal-mediated disease and development of a novel remediation system based on fieldwork and experimental research].

    PubMed

    Yajima, Ichiro; Zou, Cunchao; Li, Xiang; Nakano, Chizuru; Omata, Yasuhiro; Kumasaka, Mayuko Y

    2015-01-01

    Heavy-metal pollution occurs in various environments, including water, air and soil, and has serious effects on human health. Since heavy-metal pollution in drinking water causes various diseases including skin cancer, it has become a global problem worldwide. However, there is limited information on the mechanism of development of heavy-metal-mediated disease. We performed both fieldwork and experimental studies to elucidate the levels of heavy-metal pollution and mechanisms of development of heavy-metal-related disease and to develop a novel remediation system. Our fieldwork in Bangladesh, Vietnam and Malaysia demonstrated that drinking well water in these countries was polluted with high concentrations of several heavy metals including arsenic, barium, iron and manganese. Our experimental studies based on the data from our fieldwork demonstrated that these heavy metals caused skin cancer and hearing loss. Further experimental studies resulted in the development of a novel remediation system with which toxic heavy metals were absorbed from polluted drinking water. Implementation of both fieldwork and experimental studies is important for prediction, prevention and therapy of heavy-metal-mediated diseases.

  19. Metal-Mediated Affinity and Orientation Specificity in a Computationally Designed Protein Homodimer

    SciTech Connect

    Der, Bryan S.; Machius, Mischa; Miley, Michael J.; Mills, Jeffrey L.; Szyperski, Thomas; Kuhlman, Brian

    2015-10-15

    Computationally designing protein-protein interactions with high affinity and desired orientation is a challenging task. Incorporating metal-binding sites at the target interface may be one approach for increasing affinity and specifying the binding mode, thereby improving robustness of designed interactions for use as tools in basic research as well as in applications from biotechnology to medicine. Here we describe a Rosetta-based approach for the rational design of a protein monomer to form a zinc-mediated, symmetric homodimer. Our metal interface design, named MID1 (NESG target ID OR37), forms a tight dimer in the presence of zinc (MID1-zinc) with a dissociation constant <30 nM. Without zinc the dissociation constant is 4 {micro}M. The crystal structure of MID1-zinc shows good overall agreement with the computational model, but only three out of four designed histidines coordinate zinc. However, a histidine-to-glutamate point mutation resulted in four-coordination of zinc, and the resulting metal binding site and dimer orientation closely matches the computational model (C{alpha} rmsd = 1.4 {angstrom}).

  20. Metal-ion-mediated assemblies of thiazole orange with cucurbit[7]uril: a photophysical study.

    PubMed

    Shinde, Meenakshi N; Dutta Choudhury, Sharmistha; Barooah, Nilotpal; Pal, Haridas; Bhasikuttan, Achikanath C; Mohanty, Jyotirmayee

    2015-03-05

    The formation of molecular superstructures by metal-ion-mediated noncovalent self-assembly has been demonstrated using the macrocycle, cucurbit[7]uril (CB7), and the dye, thiazole orange (TO), as building blocks. Interestingly, the association of these molecular building blocks can be tuned by the chemical environment, leading to self-assembled structures of different stoichiometries, which is supported by absorption, fluorescence, (1)H NMR, and AFM measurements. Most importantly, the self-assembly process of the CB7/TO/metal ion system is observed to be remarkably different for alkali (Na(+)) and alkaline earth (Ca(2+)) metal ions. Fluorescence enhancement is observed in the presence of Ca(2+) ions, which is attributed to the formation of short dimeric structures composed of two 1:1 CB7-TO complexes. Solution turbidity is detected in the presence of Na(+) ions, which is proposed to be due to the formation of extended structures by the assembly of many 1:1 CB7-TO complexes.

  1. Polydopamine-mediated surface-functionalization of graphene oxide for heavy metal ions removal

    NASA Astrophysics Data System (ADS)

    Dong, Zhihui; Zhang, Feng; Wang, Dong; Liu, Xia; Jin, Jian

    2015-04-01

    By utilizing polydopamine (PD) nano-thick interlayer as mediator, polyethylenimine (PEI) brushes with abundant amine groups were grafted onto the surface of PD coated graphene oxide (GO) uniformly via a Michael-Addition reaction and produced a PEI-PD/GO composite nanosheets. The PEI-PD/GO composite exhibited an improved performance for adsorption of heavy metal ions as compared to PEI-coated GO and pure GO. The adsorption capacities for Cu2+, Cd2+, Pb2+, Hg2+ are up to 87, 106, 197, and 110 mg/g, respectively. To further make the GO based composite operable, PEI-PD/RGO aerogel was prepared through hydrothermal and achieved a high surface area up to 373 m2/g. Although the adsorption capacity of PEI-PD/RGO aerogel for heavy metal ions decreases a little as compared to PEI-PD/GO composite dispersion (38, 32, 95, 113 mg/g corresponding to Cu2+, Cd2+, Pb2+, and Hg2+, respectively), it could be recycled several times in a simple way by releasing adsorbed metal ions, indicating its potential application for cleaning wastewater.

  2. Hyperbranched polymer mediated fabrication of water soluble carbon nanotube-metal nanoparticle hybrids

    NASA Astrophysics Data System (ADS)

    Li, Haiqing; Cooper-White, Justin J.

    2013-03-01

    1-Pyrenemethanol initiated hyperbranched polyglycerol (PiHP) has been synthesized and utilized to non-covalently functionalize pristine multi-walled carbon nanotubes (CNTs) through π-π stacking interactions. Mediated with the PiHP coating, a variety of metal nanoparticles (Au, Ag, Pd and Pt) were in situ generated and randomly tethered on the CNT sidewalls, producing various water-soluble CNT/PiHP/metal hybrids. Particularly, the resulting CNT/PiHP/Pt hybrids possess improved metal coverage in comparison to the reported CNT/Pt nanohybrids obtained by the use of conventional non-covalent CNT surface-modifiers. Depending on the using concentration of Pt2+ precursor, Pt coverage in CNT/PiHP/Pt hybrids can be effectively controlled. In the meanwhile, Pt component on the CNT sidewalls can be either well isolated nanoparticles or loose ``nanoclusters''. To test the promising catalytic application of these obtained CNT/PiHP/Pt hybrids, a systematic investigation on their catalytic performance towards the reduction of 4-nitrophenol to produce 4-aminophenol was performed. Surprisingly, these hybrids exhibited significantly enhanced catalytic activity compared with the conventionally utilized Au and Ag nanoparticles. Moreover, they can be easily recovered and reused without significant loss in catalytic activity after running 6 circles.

  3. Synthetic strategies and structural aspects of metal-mediated multiporphyrin assemblies.

    PubMed

    Iengo, Elisabetta; Zangrando, Ennio; Alessio, Enzo

    2006-11-01

    Porphyrins play a major role as active chromophores in artificial systems mimicking the natural photoinduced processes. The formation of coordination bonds between peripheral donor sites on the porphyrins and external metal fragments has proven to be an efficient