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Sample records for idade da planta

  1. Synthetic promoters in planta.

    PubMed

    Dey, Nrisingha; Sarkar, Shayan; Acharya, Sefali; Maiti, Indu B

    2015-11-01

    This paper reviews the importance, prospective and development of synthetic promoters reported in planta. A review of the synthetic promoters developed in planta would help researchers utilize the available resources and design new promoters to benefit fundamental research and agricultural applications. The demand for promoters for the improvement and application of transgenic techniques in research and agricultural production is increasing. Native/naturally occurring promoters have some limitations in terms of their induction conditions, transcription efficiency and size. The strength and specificity of native promoter can be tailored by manipulating its 'cis-architecture' by the use of several recombinant DNA technologies. Newly derived chimeric promoters with specific attributes are emerging as an efficient tool for plant molecular biology. In the last three decades, synthetic promoters have been used to regulate plant gene expression. To better understand synthetic promoters, in this article, we reviewed promoter structure, the scope of cis-engineering, strategies for their development, their importance in plant biology and the total number of such promoters (188) developed in planta to date; we then categorized them under different functional regimes as biotic stress-inducible, abiotic stress-inducible, light-responsive, chemical-inducible, hormone-inducible, constitutive and tissue-specific. Furthermore, we identified a set of 36 synthetic promoters that control multiple types of expression in planta. Additionally, we illustrated the differences between native and synthetic promoters and among different synthetic promoter in each group, especially in terms of efficiency and induction conditions. As a prospective of this review, the use of ideal synthetic promoters is one of the prime requirements for generating transgenic plants suitable for promoting sustainable agriculture and plant molecular farming.

  2. Nebulosas planetárias galácticas: idades e cinemática

    NASA Astrophysics Data System (ADS)

    Lago, L. G.; Maciel, W. J.

    2003-08-01

    As nebulosas planetárias são formadas a partir de estrelas com massas entre 0.8 e 8 massas solares na sequência principal, formando um conjunto de objetos com idades e populações diferentes. Esse fato torna as nebulosas interessantes ao estudo da rotação galáctica, uma vez que as populações mais jovens devem estar claramente associadas ao disco fino da Galáxia, enquanto que as populações mais velhas estariam associadas ao disco espesso ou mesmo ao halo galáctico. Neste trabalho, utilizamos uma grande amostra de nebulosas com velocidades radiais conhecidas para determinar a curva de rotação da Galáxia. Para minimizar os erros decorrentes das incertezas nas distâncias, usamos quatro diferentes escalas estatísticas. Para a atribuição da idade dos objetos, utilizamos o método proposto recentemente por Maciel et al. (Astron. Astrophys. 397, 667, 2003), em um estudo da variação temporal dos gradientes de abundâncias. Com esse método, é possível obter uma classificação das nebulosas por grupos de idade, totalmente independente de outros sistemas de classificação, uma vez que as idades são estimadas individualmente, a partir das metalicidades das nebulosas. Nossos resultados são comparados com as previsões dos tipos de Peimbert, e mostram que os desvios médios das curvas obtidas são sensivelmente menores nos casos dos objetos mais jovens, associados ao disco fino, com relação aos objetos mais velhos do disco espesso, levando à determinação de uma curva de rotação semelhante à dos objetos de população I jovem. (CNPq)

  3. Fibroma of tendon sheath in planta.

    PubMed

    Lu, Hui; Chen, Qiang; Shen, Hui; Shen, Xiang-Qian; Wu, Shou-Cheng

    2016-01-01

    Fibroma of tendon sheath in planta is comparatively rare, and its differentiate diagnose, tumour features, treatment and complications were lack of retrospective study in clinics. This was a retrospective study of 13 patients (seven women, six men) operated between July 2001 and May 2013 for FTS in planta. The average age at the time of the procedure was 49.8 ± 8.3 years old (range 31-64). The female-to-male ratio was 9:4. Before the surgery, anteroposterior, lateral and oblique position of foot X-rays were performed in all patients. Ultrasonography (n = 11) and magnetic resonance imaging (n = 11) were performed selectively. The tumor located on the metatarsal par (n = 6), the central part of plantar (n = 4), the lateral part (n = 2) and the medial part (n = 1). Eight patients presented with painless mass (62 % of cases), while five patients presented with pain mass (38 % of cases). No patient had bony erosion. This paper studies the different features of FTS and classifies them into two types-superficial type that tumour grows at planter fascia; deep type that breaks through the planter fascia growing around tendon and joint capsule. Eight and five patients were diagnosed as superficial type and deep type respectively. In all cases, the tumor was excised, pathological results was FTS. The mean follow-up period was 3.2 ± 1.1 years (range 2-7) years. Five patients had neurovascular bundle involvement (38 % of cases). Two patients had a recurrence (15 % of cases), they undergone another operation. Four patients had a pain (31 % of cases), two patients had numbness (15 % of cases), and one patient had pain and numbness (8 % of cases). They recovered after conservative treatment. For the FTS that grows in the plantar, we should select differential diagnosis and the corresponding therapy according to the features of two types, also the prognosis is different. IV.

  4. Hyperthermophilic endoglucanase for in planta lignocellulose conversion

    PubMed Central

    2012-01-01

    Background The enzymatic conversion of lignocellulosic plant biomass into fermentable sugars is a crucial step in the sustainable and environmentally friendly production of biofuels. However, a major drawback of enzymes from mesophilic sources is their suboptimal activity under established pretreatment conditions, e.g. high temperatures, extreme pH values and high salt concentrations. Enzymes from extremophiles are better adapted to these conditions and could be produced by heterologous expression in microbes, or even directly in the plant biomass. Results Here we show that a cellulase gene (sso1354) isolated from the hyperthermophilic archaeon Sulfolobus solfataricus can be expressed in plants, and that the recombinant enzyme is biologically active and exhibits the same properties as the wild type form. Since the enzyme is inactive under normal plant growth conditions, this potentially allows its expression in plants without negative effects on growth and development, and subsequent heat-inducible activation. Furthermore we demonstrate that the recombinant enzyme acts in high concentrations of ionic liquids and can therefore degrade α-cellulose or even complex cell wall preparations under those pretreatment conditions. Conclusion The hyperthermophilic endoglucanase SSO1354 with its unique features is an excellent tool for advanced biomass conversion. Here we demonstrate its expression in planta and the possibility for post harvest activation. Moreover the enzyme is suitable for combined pretreatment and hydrolysis applications. PMID:22928996

  5. 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.

  6. In planta passive sampling devices for assessing subsurface chlorinated solvents.

    PubMed

    Shetty, Mikhil K; Limmer, Matt A; Waltermire, Kendra; Morrison, Glenn C; Burken, Joel G

    2014-06-01

    Contaminant concentrations in trees have been used to delineate groundwater contaminant plumes (i.e., phytoscreening); however, variability in tree composition hinders accurate measurement of contaminant concentrations in planta, particularly for long-term monitoring. This study investigated in planta passive sampling devices (PSDs), termed solid phase samplers (SPSs) to be used as a surrogate tree core. Characteristics studied for five materials included material-air partitioning coefficients (Kma) for chlorinated solvents, sampler equilibration time and field suitability. The materials investigated were polydimethylsiloxane (PDMS), low-density polyethylene (LDPE), linear low-density polyethylene (LLDPE), polyoxymethylene (POM) and plasticized polyvinyl chloride (PVC). Both PDMS and LLDPE samplers demonstrated high partitioning coefficients and diffusivities and were further tested in greenhouse experiments and field trials. While most of the materials could be used for passive sampling, the PDMS SPSs performed best as an in planta sampler. Such a sampler was able to accurately measure trichloroethylene (TCE) and tetrachloroethylene (PCE) concentrations while simultaneously incorporating simple operation and minimal impact to the surrounding property and environment. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Native-sized spider silk proteins synthesized in planta via intein-based multimerization.

    PubMed

    Hauptmann, Valeska; Weichert, Nicola; Menzel, Matthias; Knoch, Dominic; Paege, Norman; Scheller, Jürgen; Spohn, Uwe; Conrad, Udo; Gils, Mario

    2013-04-01

    The synthesis of native-sized proteins is a pre-requisite for exploiting the potential of spider silk as a bio-based material. The unique properties of spider silk, such as extraordinary tensile strength and elasticity, result from the highly repetitive nature of spider silk protein motifs. The present report describes the combination of spider silk flagelliform protein (FLAG) production in the endoplasmic reticulum of tobacco plant leaf cells with an intein-based posttranslational protein fusion technology. The repeated ligation of FLAG monomers resulted in the formation of large multimers. This method avoids the need for highly repetitive transgenes, which may result in a higher genetic and transcriptional stability. Here we show, for the first time, the production of synthetic, high molecular weight spider silk proteins larger than 250 kDa based on the assembly of protein monomers via intein-mediated trans-splicing in planta. The resulting multimeric structures form microfibers, thereby demonstrating their great potential as a biomaterial.

  8. Comparative proteomics of extracellular proteins in vitro and in planta from the pathogenic fungus Fusarium graminearum.

    PubMed

    Paper, Janet M; Scott-Craig, John S; Adhikari, Neil D; Cuomo, Christina A; Walton, Jonathan D

    2007-09-01

    High-throughput MS/MS was used to identify proteins secreted by Fusarium graminearum (Gibberella zeae) during growth on 13 media in vitro and in planta during infection of wheat heads. In vitro secreted proteins were collected from the culture filtrates, and in planta proteins were collected by vacuum infiltration. A total of 289 proteins (229 in vitro and 120 in planta) were identified with high statistical confidence. Forty-nine of the in planta proteins were not found in any of the in vitro conditions. The majority (91-100%) of the in vitro proteins had predicted signal peptides, but only 56% of the in planta proteins. At least 13 of the nonsecreted proteins found only in planta were single-copy housekeeping enzymes, including enolase, triose phosphate isomerase, phosphoglucomutase, calmodulin, aconitase, and malate dehydrogenase. The presence of these proteins in the in planta but not in vitro secretome might indicate that significant fungal lysis occurs during pathogenesis. On the other hand, several of the proteins lacking signal peptides that were found in planta have been reported to be potent immunogens secreted by animal pathogenic fungi, and therefore could be important in the interaction between F. graminearum and its host plants.

  9. CRISPR/Cas-Mediated In Planta Gene Targeting.

    PubMed

    Schiml, Simon; Fauser, Friedrich; Puchta, Holger

    2017-01-01

    The recent emergence of the CRISPR/Cas system has boosted the possibilities for precise genome engineering approaches throughout all kingdoms of life. The most common application for plants is targeted mutagenesis, whereby a Cas9-mediated DNA double-strand break (DSB) is repaired by mutagenic nonhomologous end joining (NHEJ). However, the site-specific alteration of a genomic sequence or integration of a transgene relies on the precise repair by homologous recombination (HR) using a suitable donor sequence: this poses a particular challenge in plants, as NHEJ is the preferred repair mechanism for DSBs in somatic tissue. Here, we describe our recently developed in planta gene targeting (ipGT) system, which works via the induction of DSBs by Cas9 to activate the target and the targeting vector at the same time, making it independent of high transformation efficiencies.

  10. The effector AvrRxo1 phosphorylates NAD in planta.

    PubMed

    Shidore, Teja; Broeckling, Corey D; Kirkwood, Jay S; Long, John J; Miao, Jiamin; Zhao, Bingyu; Leach, Jan E; Triplett, Lindsay R

    2017-06-01

    Gram-negative bacterial pathogens of plants and animals employ type III secreted effectors to suppress innate immunity. Most characterized effectors work through modification of host proteins or transcriptional regulators, although a few are known to modify small molecule targets. The Xanthomonas type III secreted avirulence factor AvrRxo1 is a structural homolog of the zeta toxin family of sugar-nucleotide kinases that suppresses bacterial growth. AvrRxo1 was recently reported to phosphorylate the central metabolite and signaling molecule NAD in vitro, suggesting that the effector might enhance bacterial virulence on plants through manipulation of primary metabolic pathways. In this study, we determine that AvrRxo1 phosphorylates NAD in planta, and that its kinase catalytic sites are necessary for its toxic and resistance-triggering phenotypes. A global metabolomics approach was used to independently identify 3'-NADP as the sole detectable product of AvrRxo1 expression in yeast and bacteria, and NAD kinase activity was confirmed in vitro. 3'-NADP accumulated upon transient expression of AvrRxo1 in Nicotiana benthamiana and in rice leaves infected with avrRxo1-expressing strains of X. oryzae. Mutation of the catalytic aspartic acid residue D193 abolished AvrRxo1 kinase activity and several phenotypes of AvrRxo1, including toxicity in yeast, bacteria, and plants, suppression of the flg22-triggered ROS burst, and ability to trigger an R gene-mediated hypersensitive response. A mutation in the Walker A ATP-binding motif abolished the toxicity of AvrRxo1, but did not abolish the 3'-NADP production, virulence enhancement, ROS suppression, or HR-triggering phenotypes of AvrRxo1. These results demonstrate that a type III effector targets the central metabolite and redox carrier NAD in planta, and that this catalytic activity is required for toxicity and suppression of the ROS burst.

  11. In Planta Localization of Stilbenes within Picea abies Phloem.

    PubMed

    Jyske, Tuula; Kuroda, Katsushi; Suuronen, Jussi-Petteri; Pranovich, Andrey; Roig-Juan, Sílvia; Aoki, Dan; Fukushima, Kazuhiko

    2016-10-01

    Phenolic stilbene glucosides (astringin, isorhapontin, and piceid) and their aglycons commonly accumulate in the phloem of Norway spruce (Picea abies). However, current knowledge about the localization and accumulation of stilbenes within plant tissues and cells remains limited. Here, we used an innovative combination of novel microanalytical techniques to evaluate stilbenes in a frozen-hydrated condition (i.e. in planta) and a freeze-dried condition across phloem tissues. Semiquantitative time-of-flight secondary ion-mass spectrometry imaging in planta revealed that stilbenes were localized in axial parenchyma cells. Quantitative gas chromatography analysis showed the highest stilbene content in the middle of collapsed phloem with decreases toward the outer phloem. The same trend was detected for soluble sugar and water contents. The specimen water content may affect stilbene composition; the glucoside-to-aglycon ratio decreased slightly with decreases in water content. Phloem chemistry was correlated with three-dimensional structures of phloem as analyzed by microtomography. The outer phloem was characterized by a high volume of empty parenchyma, reduced ray volume, and a large number of axial parenchyma with porous vacuolar contents. Increasing porosity from the inner to the outer phloem was related to decreasing compactness of stilbenes and possible secondary oxidation or polymerization. Our results indicate that aging-dependent changes in phloem may reduce cell functioning, which affects the capacity of the phloem to store water and sugar, and may reduce the defense potential of stilbenes in the axial parenchyma. Our results highlight the power of using a combination of techniques to evaluate tissue- and cell-level mechanisms involved in plant secondary metabolite formation and metabolism. © 2016 American Society of Plant Biologists. All Rights Reserved.

  12. The effector AvrRxo1 phosphorylates NAD in planta

    PubMed Central

    Long, John J.; Zhao, Bingyu; Leach, Jan E.

    2017-01-01

    Gram-negative bacterial pathogens of plants and animals employ type III secreted effectors to suppress innate immunity. Most characterized effectors work through modification of host proteins or transcriptional regulators, although a few are known to modify small molecule targets. The Xanthomonas type III secreted avirulence factor AvrRxo1 is a structural homolog of the zeta toxin family of sugar-nucleotide kinases that suppresses bacterial growth. AvrRxo1 was recently reported to phosphorylate the central metabolite and signaling molecule NAD in vitro, suggesting that the effector might enhance bacterial virulence on plants through manipulation of primary metabolic pathways. In this study, we determine that AvrRxo1 phosphorylates NAD in planta, and that its kinase catalytic sites are necessary for its toxic and resistance-triggering phenotypes. A global metabolomics approach was used to independently identify 3’-NADP as the sole detectable product of AvrRxo1 expression in yeast and bacteria, and NAD kinase activity was confirmed in vitro. 3’-NADP accumulated upon transient expression of AvrRxo1 in Nicotiana benthamiana and in rice leaves infected with avrRxo1-expressing strains of X. oryzae. Mutation of the catalytic aspartic acid residue D193 abolished AvrRxo1 kinase activity and several phenotypes of AvrRxo1, including toxicity in yeast, bacteria, and plants, suppression of the flg22-triggered ROS burst, and ability to trigger an R gene-mediated hypersensitive response. A mutation in the Walker A ATP-binding motif abolished the toxicity of AvrRxo1, but did not abolish the 3’-NADP production, virulence enhancement, ROS suppression, or HR-triggering phenotypes of AvrRxo1. These results demonstrate that a type III effector targets the central metabolite and redox carrier NAD in planta, and that this catalytic activity is required for toxicity and suppression of the ROS burst. PMID:28628666

  13. Construção de um catálogo de aglomerados abertos para estudo da dinâmica da estrutura espiral da Galáxia

    NASA Astrophysics Data System (ADS)

    Carlos, I. M.; Lépine, J. R. D.

    2003-08-01

    Os aglomerados abertos são objetos de grande valor para o estudo da dinâmica da Galáxia devido esses objetos terem uma faixa de idade relativamente ampla. O trabalho visa estudar a dinâmica da estrutura espiral da Galáxia principalmente através do uso desses aglomerados, uma vez que o estudo da cinemática desses objetos é fundamental para esse objetivo. Nosso grupo trabalha no sentido de construir uma base de dados de aglomerados abertos contendo coordenadas, distância, idade, movimentos próprios e velocidades radiais e já disponibiliza uma nova versão do catálogo de aglomerados abertos o qual é uma compilação de edições anteriores, principalmente Lynga (1987), Mermilliod (1995) e ESO-B (Lauberts 1982). Nossa amostra possui cerca de 1630 aglomerados, mas nem todos os parâmetros acima citados foram determinados em sua totalidade. Para determinarmos esses parâmetros, derivamos as cores intrínsecas das estrelas membro de cada aglomerado a partir de seus tipos espectrais (busca feita no SIMBAD) obtendo assim o excesso de cor individual. A distribuição dos excessos de cor foi então utilizada para derivarmos o avermelhamento médio para cada aglomerado. De maneira similar, os tipos espectrais foram usados para estimar as magnitudes absolutas, e com as magnitudes absolutas e aparentes determinamos a respectiva distribuição do módulo de distância e finalmente a distância. Para determinar as idades foram confeccionados os diagramas cor-magnitude das estrelas de cada aglomerado onde foram superpostas a Seqüência Principal de Idade Zero (ZAMS). Superpomos a ZAMS de Schmidt-Kaler e isócronas de composição solar. Essas isócronas foram usadas para determinação das idades dos aglomerados. Uma vez que não temos ainda resultados finais, apresentamos então alguns diagramas cor-magnitude os quais foram usados para determinação, principalmente, da distância e idade dos aglomerados.

  14. Physiology of Ex Planta Nitrogenase Activity in Rhizobium japonicum†

    PubMed Central

    Agarwal, Arun K.; Keister, Donald L.

    1983-01-01

    Thirty-nine wild-type strains of Rhizobium japonicum have been studied for their ability to synthesize nitrogenase ex planta in defined liquid media under microaerobic conditions. Twenty-one produced more than trace amounts of acetylene reduction activity, but only a few of these yielded high activity. The oxygen response curves were similar for most of the nitrogenase-positive strains. The strains derepressible for activity had several phenotypic characteristics different from non-derepressible strains. These included slower growth and lower oxygen consumption under microaerobic conditions and lower extracellular polysaccharide production. Extracellular polysaccharide production during growth on gluconate in every nitrogenase-positive strain assayed was lower under both aerobic and microaerobic conditions than the non-derepressible strains. These phenotypic characteristics may be representative of a genotype of a subspecies of R. japonicum. These studies were done in part to enlarge the base number of strains available for studies on the physiology, biochemistry, and genetics of nitrogen fixation. PMID:16346295

  15. Pentahydroxyscirpene—Producing Strains, Formation In Planta, and Natural Occurrence

    PubMed Central

    Varga, Elisabeth; Wiesenberger, Gerlinde; Fruhmann, Philipp; Malachová, Alexandra; Svoboda, Thomas; Lemmens, Marc; Adam, Gerhard; Berthiller, Franz

    2016-01-01

    Trichothecenes are a class of structurally diverse mycotoxins with more than 200 naturally occurring compounds. Previously, a new compound, pentahydroxyscirpene (PHS), was reported as a byproduct of a nivalenol producing Fusarium strain, IFA189. PHS contains a hydroxy group at C-8 instead of the keto group of type B trichothecenes. In this work, we demonstrate that IFA189 belongs to the species Fusarium kyushuense using molecular tools. Production of PHS in vitro was also observed for several isolates of other Fusarium species producing nivalenol. Furthermore, we report the formation of 4-acetyl-PHS by F. kyushuense on inoculated rice. Wheat ears of the variety Remus were infected with IFA189 and the in planta production of PHS was confirmed. Natural occurrence of PHS was verified in barley samples from the Czech Republic using a liquid chromatographic-tandem mass spectrometric method validated for this purpose. Toxicity of PHS to wheat ribosomes was evaluated with a coupled in vitro transcription and translation assay, which showed that PHS inhibits protein biosynthesis slightly less than nivalenol and deoxynivalenol. PMID:27754401

  16. Physiology of ex planta nitrogenase activity in Rhizobium japonicum

    SciTech Connect

    Agarwal, A.K.; Keister, D.L.

    1983-05-01

    Thirty-nine wild-type strains of Rhizobium japonicum have been studied for their ability to synthesize nitrogenase ex planta in defined liquid media under microaerobic conditions. Twenty-one produced more than trace amounts of acetylene reduction activity, but only a few of these yielded high activity. The oxygen response curves were similar for most of the nitrogenase-positive strains. The strains derepressible for activity had several phenotypic characteristics different from non-derepressible strains. These included slower growth and lower oxygen consumption under microaerobic conditions and lower extracellular polysaccharide production. Extracellular polysaccharide production during growth on gluconate in every nitrogenase-positive strain assayed was lower under both aerobic and microaerobic conditions than the non-depressible strains. These phenotypic characteristics may be representative of a genotype of a subspecies of R. japonicum. These studies were done in part to enlarge the base number of strains available for studies on the physiology, biochemistry, and genetics of nitrogen fixation. (35 Refs.)

  17. Smallpox subunit vaccine produced in planta confers protection in mice

    PubMed Central

    Golovkin, Maxim; Spitsin, Sergei; Andrianov, Vyacheslav; Smirnov, Yuriy; Xiao, Yuhong; Pogrebnyak, Natalia; Markley, Karen; Brodzik, Robert; Gleba, Yuri; Isaacs, Stuart N.; Koprowski, Hilary

    2007-01-01

    We report here the in planta production of the recombinant vaccinia virus B5 antigenic domain (pB5), an attractive component of a subunit vaccine against smallpox. The antigenic domain was expressed by using efficient transient and constitutive plant expression systems and tested by various immunization routes in two animal models. Whereas oral administration in mice or the minipig with collard-derived insoluble pB5 did not generate an anti-B5 immune response, intranasal administration of soluble pB5 led to a rise of B5-specific immunoglobulins, and parenteral immunization led to a strong anti-B5 immune response in both mice and the minipig. Mice immunized i.m. with pB5 generated an antibody response that reduced virus spread in vitro and conferred protection from challenge with a lethal dose of vaccinia virus. These results indicate the feasibility of producing safe and inexpensive subunit vaccines by using plant production systems. PMID:17428917

  18. Pentahydroxyscirpene-Producing Strains, Formation In Planta, and Natural Occurrence.

    PubMed

    Varga, Elisabeth; Wiesenberger, Gerlinde; Fruhmann, Philipp; Malachová, Alexandra; Svoboda, Thomas; Lemmens, Marc; Adam, Gerhard; Berthiller, Franz

    2016-10-14

    Trichothecenes are a class of structurally diverse mycotoxins with more than 200 naturally occurring compounds. Previously, a new compound, pentahydroxyscirpene (PHS), was reported as a byproduct of a nivalenol producing Fusarium strain, IFA189. PHS contains a hydroxy group at C-8 instead of the keto group of type B trichothecenes. In this work, we demonstrate that IFA189 belongs to the species Fusarium kyushuense using molecular tools. Production of PHS in vitro was also observed for several isolates of other Fusarium species producing nivalenol. Furthermore, we report the formation of 4-acetyl-PHS by F. kyushuense on inoculated rice. Wheat ears of the variety Remus were infected with IFA189 and the in planta production of PHS was confirmed. Natural occurrence of PHS was verified in barley samples from the Czech Republic using a liquid chromatographic-tandem mass spectrometric method validated for this purpose. Toxicity of PHS to wheat ribosomes was evaluated with a coupled in vitro transcription and translation assay, which showed that PHS inhibits protein biosynthesis slightly less than nivalenol and deoxynivalenol.

  19. An in planta biolistic method for stable wheat transformation.

    PubMed

    Hamada, Haruyasu; Linghu, Qianyan; Nagira, Yozo; Miki, Ryuji; Taoka, Naoaki; Imai, Ryozo

    2017-09-13

    The currently favoured method for wheat (Triticum aestivum L.) transformation is inapplicable to many elite cultivars because it requires callus culture and regeneration. Here, we developed a simple, reproducible, in planta wheat transformation method using biolistic DNA delivery without callus culture or regeneration. Shoot apical meristems (SAMs) grown from dry imbibed seeds were exposed under a microscope and subjected to bombardment with different-sized gold particles coated with the GFP gene construct, introducing DNA into the L2 cell layer. Bombarded embryos were grown to mature, stably transformed T0 plants and integration of the GFP gene into the genome was determined at the fifth leaf. Use of 0.6-µm particles and 1350-psi pressure resulted in dramatically increased maximum ratios of transient GFP expression in SAMs and transgene integration in the fifth leaf. The transgene was integrated into the germ cells of 62% of transformants, and was therefore inherited in the next generation. We successfully transformed the model wheat cultivar 'Fielder', as well as the recalcitrant Japanese elite cultivar 'Haruyokoi'. Our method could potentially be used to generate stable transgenic lines for a wide range of commercial wheat cultivars.

  20. Pro-insecticidal approach towards increasing in planta activity.

    PubMed

    Creemer, Lawrence C; Giampietro, Natalie C; Lambert, William; Yap, Maurice C; deBoer, Gerrit J; Adelfinskaya, Yelena; Castetter, Scott; Wessels, Frank J

    2017-04-01

    The adrenergic mode of action was investigated for the development of potential new insecticides. Clonidine-related analogs were tested against Myzus persicae (Sulzer) and Bemisia tabaci (Gennadius). Clonidine analogs lack translation owing to a possible vacuole-trapping mechanism. Physical property modulation via a prodrug approach was attempted to overcome this mechanism. Clonidine showed insecticidal activity against M. persicae and B. tabaci. A prodrug of a known open-chain analog of clonidine was developed. While the prodrug had decreased pKa and increased lipophilicity and displayed good activity against M. persicae B. tabaci, the activity did not translate to cotton. Metabolic studies showed that the prodrug was quickly metabolized to the parent compound, and was further metabolized to a known vacuole-trapped oxazoline analog. Adrenergic active compounds, such as clonidine analogs, show potential as insecticides; however, a designed prodrug approach did not overcome the lack of translation in this case. Studies confirmed that the synthesized prodrug analog metabolized in planta to the proposed vacuole-trapped compound. One possible explanation for the failure of this approach is that the rate of metabolism and vacuole trapping is faster than translaminar flow, and therefore the released pesticide is not biologically available to the target organism. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  1. Time Gating of Chloroplast Autofluorescence Allows Clearer Fluorescence Imaging In Planta

    PubMed Central

    Kodama, Yutaka

    2016-01-01

    Chloroplast, an organelle facilitating photosynthesis, exhibits strong autofluorescence, which is an undesired background signal that restricts imaging experiments with exogenous fluorophore in plants. In this study, the autofluorescence was characterized in planta under confocal laser microscopy, and it was found that the time-gated imaging technique completely eliminates the autofluorescence. As a demonstration of the technique, a clearer signal of fluorescent protein-tagged phototropin, a blue-light photoreceptor localized at the chloroplast periphery, was visualized in planta. PMID:27027881

  2. Time Gating of Chloroplast Autofluorescence Allows Clearer Fluorescence Imaging In Planta.

    PubMed

    Kodama, Yutaka

    2016-01-01

    Chloroplast, an organelle facilitating photosynthesis, exhibits strong autofluorescence, which is an undesired background signal that restricts imaging experiments with exogenous fluorophore in plants. In this study, the autofluorescence was characterized in planta under confocal laser microscopy, and it was found that the time-gated imaging technique completely eliminates the autofluorescence. As a demonstration of the technique, a clearer signal of fluorescent protein-tagged phototropin, a blue-light photoreceptor localized at the chloroplast periphery, was visualized in planta.

  3. Antifreeze Proteins Modify the Freezing Process In Planta12

    PubMed Central

    Griffith, Marilyn; Lumb, Chelsey; Wiseman, Steven B.; Wisniewski, Michael; Johnson, Robert W.; Marangoni, Alejandro G.

    2005-01-01

    During cold acclimation, winter rye (Secale cereale L. cv Musketeer) plants accumulate antifreeze proteins (AFPs) in the apoplast of leaves and crowns. The goal of this study was to determine whether these AFPs influence survival at subzero temperatures by modifying the freezing process or by acting as cryoprotectants. In order to inhibit the growth of ice, AFPs must be mobile so that they can bind to specific sites on the ice crystal lattice. Guttate obtained from cold-acclimated winter rye leaves exhibited antifreeze activity, indicating that the AFPs are free in solution. Infrared video thermography was used to observe freezing in winter rye leaves. In the absence of an ice nucleator, AFPs had no effect on the supercooling temperature of the leaves. However, in the presence of an ice nucleator, AFPs lowered the temperature at which the leaves froze by 0.3°C to 1.2°C. In vitro studies showed that apoplastic proteins extracted from cold-acclimated winter rye leaves inhibited the recrystallization of ice and also slowed the rate of migration of ice through solution-saturated filter paper. When we examined the possible role of winter rye AFPs in cryoprotection, we found that lactate dehydrogenase activity was higher after freezing in the presence of AFPs compared with buffer, but the same effect was obtained by adding bovine serum albumin. AFPs had no effect on unstacked thylakoid volume after freezing, but did inhibit stacking of the thylakoids, thus indicating a loss of thylakoid function. We conclude that rye AFPs have no specific cryoprotective activity; rather, they interact directly with ice in planta and reduce freezing injury by slowing the growth and recrystallization of ice. PMID:15805474

  4. Secretome analysis of the rice bacterium Xanthomonas oryzae (Xoo) using in vitro and in planta systems.

    PubMed

    Wang, Yiming; Kim, Sang Gon; Wu, Jingni; Huh, Hyun-Hye; Lee, Su-Ji; Rakwal, Randeep; Agrawal, Ganesh Kumar; Park, Zee-Yong; Young Kang, Kyu; Kim, Sun Tae

    2013-06-01

    Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight disease in rice, and that severely affects yield loss (upto 50%) of total rice production. Here, we report a proteomics investigation of Xoo (compatible race K3)-secreted proteins, isolated from its in vitro culture and in planta infected rice leaves. 2DE coupled with MALDI-TOF-MS and/or nLC-ESI-MS/MS approaches identified 139 protein spots (out of 153 differential spots), encoding 109 unique proteins. Identified proteins belonged to multiple biological and molecular functions. Metabolic and nutrient uptake proteins were common up to both in vitro and in planta secretomes. However, pathogenicity, protease/peptidase, and host defense-related proteins were highly or specifically expressed during in planta infection. A good correlation was observed between protein and transcript abundances for nine proteins secreted in planta as per semiquantitative RT-PCR analysis. Transgenic rice leaf sheath (carrying PBZ1 promoter::GFP cell death reporter), when used to express a few of the identified secretory proteins, showed a direct activation of cell death signaling, suggesting their involvement in pathogenicity related with secretion effectors. This work furthers our understanding of rice bacterial blight disease, and serves as a resource for possible translation in generating disease resistant rice plants for improved seed yield. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. A full-length infectious clone of beet soil-borne virus indicates the dispensability of the RNA-2 for virus survival in planta and symptom expression on Chenopodium quinoa leaves.

    PubMed

    Crutzen, François; Mehrvar, Mohsen; Gilmer, David; Bragard, Claude

    2009-12-01

    For a better understanding of the functionality and pathogenicity of beet soil-borne virus (BSBV), full-length cDNA clones have been constructed for the three genomic RNAs. With the aim of assessing their effectiveness and relative contribution to the virus housekeeping functions, transcripts were inoculated on Chenopodium quinoa and Beta macrocarpa leaves using five genome combinations. Both RNAs-1 (putative replicase) and -3 (putative movement proteins) proved to be essential for virus replication in planta and symptom production on C. quinoa, whereas RNA-2 (putative coat protein, CP, and a read-through domain, RT) was not. No symptoms were recorded on B. macrocarpa, but viral RNAs were detected. In both host plants, the 19 kDa CP was detected by Western blotting as well as a 115 kDa protein corresponding to the CP-RT.

  6. Chlamydiae has contributed at least 55 genes to Plantae with predominantly plastid functions.

    PubMed

    Moustafa, Ahmed; Reyes-Prieto, Adrian; Bhattacharya, Debashish

    2008-05-21

    The photosynthetic organelle (plastid) originated via primary endosymbiosis in which a phagotrophic protist captured and harnessed a cyanobacterium. The plastid was inherited by the common ancestor of the red, green (including land plants), and glaucophyte algae (together, the Plantae). Despite the critical importance of primary plastid endosymbiosis, its ancient derivation has left behind very few "footprints" of early key events in organelle genesis. To gain insights into this process, we conducted an in-depth phylogenomic analysis of genomic data (nuclear proteins) from 17 Plantae species to identify genes of a surprising provenance in these taxa, Chlamydiae bacteria. Previous studies show that Chlamydiae contributed many genes (at least 21 in one study) to Plantae that primarily have plastid functions and were postulated to have played a fundamental role in organelle evolution. Using our comprehensive approach, we identify at least 55 Chlamydiae-derived genes in algae and plants, of which 67% (37/55) are putatively plastid targeted and at least 3 have mitochondrial functions. The remainder of the proteins does not contain a bioinformatically predicted organelle import signal although one has an N-terminal extension in comparison to the Chlamydiae homolog. Our data suggest that environmental Chlamydiae were significant contributors to early Plantae genomes that extend beyond plastid metabolism. The chlamydial gene distribution and protein tree topologies provide evidence for both endosymbiotic gene transfer and a horizontal gene transfer ratchet driven by recurrent endoparasitism as explanations for gene origin. Our findings paint a more complex picture of gene origin than can easily be explained by endosymbiotic gene transfer from an organelle-like point source. These data significantly extend the genomic impact of Chlamydiae on Plantae and show that about one-half (30/55) of the transferred genes are most closely related to sequences emanating from the genome

  7. Assessment of fluorescein-based fluorescent dyes for tracing Neotyphodium endophytes in planta.

    PubMed

    Card, Stuart D; Tapper, Brian A; Lloyd-West, Catherine; Wright, Kathryn M

    2013-01-01

    Fluorescent dyes were assessed for their ability to stain viable hyphae of the fungi Neotyphodium lolii and N. coenophialum, symbiotic endophytes of the Pooideae grasses Lolium perenne and Festuca arundinacea, respectively. The fluorescein-based fluorophores; fluorescein diacetate (FDA), 5(6)-carboxy-fluorescein diacetate (CFDA), 5-chloromethylfluorescein diacetate (CMFDA) and the chitin-binding stain, Calcofluor while M2R, were assessed for staining of endophyte hyphae in vitro from axenic fungal cultures and in planta, including epidermal leaf sheath peels, nodes, ovaries, embryos and meristems. CMFDA produced the greatest intensity of staining of fungal hyphae and gave excellent contrast in planta compared to the plant cells. Compared to the other dyes, CMFDA was also the least affected by photo bleaching and continued to fluoresce up to 2 h after initial excitation. None of the fluorescent dyes stained fungal hyphae in seed.

  8. In planta mutagenesis determines the functional regions of the wheat puroindoline proteins.

    PubMed

    Feiz, L; Beecher, B S; Martin, J M; Giroux, M J

    2009-11-01

    In planta analysis of protein function in a crop plant could lead to improvements in understanding protein structure/function relationships as well as selective agronomic or end product quality improvements. The requirements for successful in planta analysis are a high mutation rate, an efficient screening method, and a trait with high heritability. Two ideal targets for functional analysis are the Puroindoline a and Puroindoline b (Pina and Pinb, respectively) genes, which together compose the wheat (Triticum aestivum L.) Ha locus that controls grain texture and many wheat end-use properties. Puroindolines (PINs) together impart soft texture, and mutations in either PIN result in hard seed texture. Studies of the PINs' mode of action are limited by low allelic variation. To create new Pin alleles and identify critical function-determining regions, Pin point mutations were created in planta via EMS treatment of a soft wheat. Grain hardness of 46 unique PIN missense alleles was then measured using segregating F(2):F(3) populations. The impact of individual missense alleles upon PIN function, as measured by grain hardness, ranged from neutral (74%) to intermediate to function abolishing. The percentage of function-abolishing mutations among mutations occurring in both PINA and PINB was higher for PINB, indicating that PINB is more critical to overall Ha function. This is contrary to expectations in that PINB is not as well conserved as PINA. All function-abolishing mutations resulted from structure-disrupting mutations or from missense mutations occurring near the Tryptophan-rich region. This study demonstrates the feasibility of in planta functional analysis of wheat proteins and that the Tryptophan-rich region is the most important region of both PINA and PINB.

  9. Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry

    PubMed Central

    2012-01-01

    Understanding protein and gene function requires identifying interaction partners using biochemical, molecular or genetic tools. In plants, searching for novel protein-protein interactions is limited to protein purification assays, heterologous in vivo systems such as the yeast-two-hybrid or mutant screens. Ideally one would be able to search for novel protein partners in living plant cells. We demonstrate that it is possible to screen for novel protein-protein interactions from a random library in protoplasted Arabidopsis plant cells and recover some of the interacting partners. Our screen is based on capturing the bi-molecular complementation of mYFP between an YN-bait fusion partner and a completely random prey YC-cDNA library with FACS. The candidate interactions were confirmed using in planta BiFC assays and in planta FRET-FLIM assays. From this work, we show that the well characterized protein Calcium Dependent Protein Kinase 3 (CPK3) interacts with APX3, HMGB5, ORP2A and a ricin B-related lectin domain containing protein At2g39050. This is one of the first randomin planta screens to be successfully employed. PMID:22789293

  10. Discovery of Phytophthora infestans Genes Expressed in Planta through Mining of cDNA Libraries

    PubMed Central

    Chaves, Diego; Pinzón, Andrés; Grajales, Alejandro; Rojas, Alejandro; Mutis, Gabriel; Cárdenas, Martha; Burbano, Daniel; Jiménez, Pedro; Bernal, Adriana; Restrepo, Silvia

    2010-01-01

    Background Phytophthora infestans (Mont.) de Bary causes late blight of potato and tomato, and has a broad host range within the Solanaceae family. Most studies of the Phytophthora – Solanum pathosystem have focused on gene expression in the host and have not analyzed pathogen gene expression in planta. Methodology/Principal Findings We describe in detail an in silico approach to mine ESTs from inoculated host plants deposited in a database in order to identify particular pathogen sequences associated with disease. We identified candidate effector genes through mining of 22,795 ESTs corresponding to P. infestans cDNA libraries in compatible and incompatible interactions with hosts from the Solanaceae family. Conclusions/Significance We annotated genes of P. infestans expressed in planta associated with late blight using different approaches and assigned putative functions to 373 out of the 501 sequences found in the P. infestans genome draft, including putative secreted proteins, domains associated with pathogenicity and poorly characterized proteins ideal for further experimental studies. Our study provides a methodology for analyzing cDNA libraries and provides an understanding of the plant – oomycete pathosystems that is independent of the host, condition, or type of sample by identifying genes of the pathogen expressed in planta. PMID:20352100

  11. Time-weighted average SPME analysis for in planta determination of cVOCs.

    PubMed

    Sheehan, Emily M; Limmer, Matt A; Mayer, Philipp; Karlson, Ulrich Gosewinkel; Burken, Joel G

    2012-03-20

    The potential of phytoscreening for plume delineation at contaminated sites has promoted interest in innovative, sensitive contaminant sampling techniques. Solid-phase microextraction (SPME) methods have been developed, offering quick, undemanding, noninvasive sampling without the use of solvents. In this study, time-weighted average SPME (TWA-SPME) sampling was evaluated for in planta quantification of chlorinated solvents. TWA-SPME was found to have increased sensitivity over headspace and equilibrium SPME sampling. Using a variety of chlorinated solvents and a polydimethylsiloxane/carboxen (PDMS/CAR) SPME fiber, most compounds exhibited near linear or linear uptake over the sampling period. Smaller, less hydrophobic compounds exhibited more nonlinearity than larger, more hydrophobic molecules. Using a specifically designed in planta sampler, field sampling was conducted at a site contaminated with chlorinated solvents. Sampling with TWA-SPME produced instrument responses ranging from 5 to over 200 times higher than headspace tree core sampling. This work demonstrates that TWA-SPME can be used for in planta detection of a broad range of chlorinated solvents and methods can likely be applied to other volatile and semivolatile organic compounds.

  12. In planta anthocyanin degradation by a vacuolar class III peroxidase in Brunfelsia calycina flowers.

    PubMed

    Zipor, Gadi; Duarte, Patrícia; Carqueijeiro, Inês; Shahar, Liat; Ovadia, Rinat; Teper-Bamnolker, Paula; Eshel, Dani; Levin, Yishai; Doron-Faigenboim, Adi; Sottomayor, Mariana; Oren-Shamir, Michal

    2015-01-01

    In contrast to detailed knowledge regarding the biosynthesis of anthocyanins, the largest group of plant pigments, little is known about their in planta degradation. It has been suggested that anthocyanin degradation is enzymatically controlled and induced when beneficial to the plant. Here we investigated the enzymatic process in Brunfelsia calycina flowers, as they changed color from purple to white. We characterized the enzymatic process by which B. calycina protein extracts degrade anthocyanins. A candidate peroxidase was partially purified and characterized and its intracellular localization was determined. The transcript sequence of this peroxidase was fully identified. A basic peroxidase, BcPrx01, is responsible for the in planta degradation of anthocyanins in B. calycina flowers. BcPrx01 has the ability to degrade complex anthocyanins, it co-localizes with these pigments in the vacuoles of petals, and both the mRNA and protein levels of BcPrx01 are greatly induced parallel to the degradation of anthocyanins. Both isoelectric focusing (IEF) gel analysis and 3D structure prediction indicated that BcPrx01 is cationic. Identification of BcPrx01 is a significant breakthrough both in the understanding of anthocyanin catabolism in plants and in the field of peroxidases, where such a consistent relationship between expression levels, in planta subcellular localization and activity has seldom been demonstrated. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  13. Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry.

    PubMed

    Berendzen, Kenneth Wayne; Böhmer, Maik; Wallmeroth, Niklas; Peter, Sébastien; Vesić, Marko; Zhou, Ying; Tiesler, Franziska Katharina Elisabeth; Schleifenbaum, Frank; Harter, Klaus

    2012-07-12

    Understanding protein and gene function requires identifying interaction partners using biochemical, molecular or genetic tools. In plants, searching for novel protein-protein interactions is limited to protein purification assays, heterologous in vivo systems such as the yeast-two-hybrid or mutant screens. Ideally one would be able to search for novel protein partners in living plant cells. We demonstrate that it is possible to screen for novel protein-protein interactions from a random library in protoplasted Arabidopsis plant cells and recover some of the interacting partners. Our screen is based on capturing the bi-molecular complementation of mYFP between an YN-bait fusion partner and a completely random prey YC-cDNA library with FACS. The candidate interactions were confirmed using in planta BiFC assays and in planta FRET-FLIM assays. From this work, we show that the well characterized protein Calcium Dependent Protein Kinase 3 (CPK3) interacts with APX3, HMGB5, ORP2A and a ricin B-related lectin domain containing protein At2g39050. This is one of the first randomin planta screens to be successfully employed.

  14. 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.

  15. Studying Protein-Protein Interactions In Planta Using Advanced Fluorescence Microscopy.

    PubMed

    Somssich, Marc; Simon, Rüdiger

    2017-01-01

    The formation of protein complexes through direct protein-protein interaction is essential for virtually every biological process, and accordingly the ability to determine the interaction properties of specific proteins is important to understand these processes. Förster resonance energy transfer (FRET) measurements are state-of-the-art confocal fluorescence microscopy- and imaging-based techniques that allow the analysis of protein interactions in vivo and in planta, in specific compartments of single cells or tissues. Here we provide a step-by-step guide to perform FRET measurements by acceptor photobleaching (APB) and fluorescence lifetime imaging microscopy (FLIM) in the plant expression system Nicotiana benthamiana.

  16. In planta cloning of geminiviral DNA: the true Sida micrantha mosaic virus.

    PubMed

    Jeske, Holger; Gotthardt, Diether; Kober, Sigrid

    2010-02-01

    The circular single-stranded DNAs of geminiviruses are multiplied efficiently and preferentially by rolling circle amplification (RCA), and can be diagnosed readily by restriction fragment length polymorphism (RFLP) and direct sequencing of the RCA product. Two strategies are described for cloning geminiviruses from plants harboring mixed infections by using RCA and RFLP with plant-derived nucleic acids without the need for bacterial amplification. By combining both these approaches, the true Sida micrantha mosaic virus was identified. The advantages of maintaining the quasispecies nature of a virus during in planta cloning is discussed with respect to reliable virus identification and resistance breeding.

  17. The alternative sigma factor AlgT, but not alginate synthesis, promotes in planta multiplication of Pseudomonas syringae pv. glycinea.

    PubMed

    Schenk, Alexander; Weingart, Helge; Ullrich, Matthias S

    2008-02-01

    The phytopathogen Pseudomonas syringae pv. glycinea produces the exopolysaccharide (EPS) alginate, which is thought to function in epiphytic fitness and virulence. A key regulator for alginate biosynthesis in Pseudomonas aeruginosa and P. syringae is the alternative sigma factor AlgT (sigma(22)). In this study, the contribution of alginate synthesis and AlgT to in planta epiphytic fitness and virulence of P. syringae was examined. Alginate biosynthesis mutants were generated for the P. syringae pv. glycinea strains PG4180 and PG4180.muc, representing a comprehensive set of alginate- and AlgT-positive or -negative derivatives. Analysis of in vitro and in planta phenotypes revealed that AlgT strongly promoted in planta growth, survival and symptom development, but decreased the ability to grow in vitro. In contrast, alginate biosynthesis had only marginal impact. Quantitative in vitro and in planta gene expression analyses for alginate biosynthesis and algT were carried out at two temperatures in AlgT-negative and -positive backgrounds. algT as well as algD gene expression was AlgT-dependent, plant-inducible and temperature-dependent, with higher expression at 18 compared to 28 degrees C; however, no temperature dependence was observed in vitro. Our data suggest that AlgT may act as a global regulator for virulence and in planta fitness traits of P. syringae independent of its role in EPS biosynthesis.

  18. i Planta

    USDA-ARS?s Scientific Manuscript database

    This application was designed for farmers, agriculture researchers, and gardeners. The application includes a calculator, a data logger, and a GPS mapping function. It was designed for vegetable farming but can be used for open field applications. The calculator solves for the amount of pesticide,...

  19. A method for obtaining RNA from Hemileia vastatrix appressoria produced in planta, suitable for transcriptomic analyses.

    PubMed

    Loureiro, Andreia; Gil Azinheira, Helena; do Céu Silva, Maria; Talhinhas, Pedro

    2015-11-01

    Appressoria are the first infection structures developed by rust fungi and require specific topographic signals from the host for their differentiation. The ease in obtaining appressoria in vitro for these biotrophic fungi led to studies concerning gene expression and gene discovery at appressorial level, avoiding the need to distinguish plant and fungal transcripts. However, in some pathosystems, it was observed that gene expression in appressoria seems to be influenced by host-derived signals, suggesting that transcriptomic analyses performed from in planta differentiated appressoria would be potentially more informative than those from in vitro differentiated appressoria. Nevertheless analysing appressorial RNA obtained from in planta samples is often hampered by an excessive dilution of fungal RNA within plant RNA, besides uncertainty regarding the fungal or plant origin of RNA from highly conserved genes. To circumvent these difficulties, we have recovered Hemileia vastatrix appressoria from Arabica coffee leaf surface using a film of nitrocellulose dissolved in butyl and ethyl acetates (nail polish), and extracted fungal RNA from the polish peel. RNA thus obtained is of good quality and usable for cDNA synthesis and transcriptomic (quantitative PCR) studies. This method could provide the means to investigate specific host-induced appressoria-related fungal pathogenicity factors.

  20. In-planta Sporulation Capacity Enhances Infectivity and Rhizospheric Competitiveness of Frankia Strains.

    PubMed

    Cotin-Galvan, Laetitia; Pozzi, Adrien C; Schwob, Guillaume; Fournier, Pascale; Fernandez, Maria P; Herrera-Belaroussi, Aude

    2016-01-01

    Frankia Sp+ strains maintain their ability to sporulate in symbiosis with actinorhizal plants, producing abundant sporangia inside host plant cells, in contrast to Sp- strains, which are unable to perform in-planta sporulation. We herein examined the role of in-planta sporulation in Frankia infectivity and competitiveness for root infection. Fifteen strains belonging to different Sp+ and Sp- phylogenetic lineages were inoculated on seedlings of Alnus glutinosa (Ag) and A. incana (Ai). Strain competitiveness was investigated by performing Sp-/Sp+ co-inoculations. Plant inoculations were standardized using crushed nodules obtained under laboratory-controlled conditions (same plant species, age, and environmental factors). Specific oligonucleotide primers were developed to identify Frankia Sp+ and/or Sp- strains in the resulting nodules. Single inoculation experiments showed that (i) infectivity by Sp+ strains was significantly greater than that by Sp- strains, (ii) genetically divergent Sp+ strains exhibited different infective abilities, and (iii) Sp+ and Sp- strains showed different host preferences according to the origin (host species) of the inocula. Co-inoculations of Sp+ and Sp- strains revealed the greater competitiveness of Sp+ strains (98.3 to 100% of Sp+ nodules, with up to 15.6% nodules containing both Sp+ and Sp- strains). The results of the present study highlight differences in Sp+/Sp- strain ecological behaviors and provide new insights to strengthen the obligate symbiont hypothesis for Sp+ strains.

  1. Ethylene Production by Pseudomonas syringae Pathovars In Vitro and In Planta

    PubMed Central

    Weingart, H.; Volksch, B.

    1997-01-01

    Significant amounts of ethylene were produced by Pseudomonas syringae pv. glycinea, pv. phaseolicola (which had been isolated from viny weed Pueraria lobata [Willd.] Ohwi [common name, kudzu]), and pv. pisi in synthetic medium. On the other hand, the bean strains of P. syringae pv. phaseolicola and strains of 17 other pathovars did not produce ethylene. P. syringae pv. glycinea and P. syringae pv. phaseolicola produced nearly identical levels of ethylene (about 5 x 10(sup-7) nl h(sup-1) cell(sup-1)), which were about 10 times higher than the ethylene level of P. syringae pv. pisi. Two 22-bp oligonucleotide primers derived from the ethylene-forming enzyme (efe) gene of P. syringae pv. phaseolicola PK2 were investigated for their ability to detect ethylene-producing P. syringae strains by PCR analysis. PCR amplification with this primer set resulted in a specific 0.99-kb fragment in all ethylene-producing strains with the exception of the P. syringae pv. pisi strains. Therefore, P. syringae pv. pisi may use a different biosynthetic pathway for ethylene production or the sequence of the efe gene is less conserved in this bacterium. P. syringae pv. phaseolicola isolated from kudzu and P. syringae pv. glycinea also produced ethylene in planta. It could be shown that the enhanced ethylene production in diseased tissue was due to the production of ethylene by the inoculated bacteria. Ethylene production in vitro and in planta was strictly growth associated. PMID:16535480

  2. Inhibition of fungal and bacterial plant pathogens in vitro and in planta with ultrashort cationic lipopeptides.

    PubMed

    Makovitzki, Arik; Viterbo, Ada; Brotman, Yariv; Chet, Ilan; Shai, Yechiel

    2007-10-01

    Plant diseases constitute an emerging threat to global food security. Many of the currently available antimicrobial agents for agriculture are highly toxic and nonbiodegradable and cause extended environmental pollution. Moreover, an increasing number of phytopathogens develop resistance to them. Recently, we have reported on a new family of ultrashort antimicrobial lipopeptides which are composed of only four amino acids linked to fatty acids (A. Makovitzki, D. Avrahami, and Y. Shai, Proc. Natl. Acad. Sci. USA 103:15997-16002, 2006). Here, we investigated the activities in vitro and in planta and the modes of action of these short lipopeptides against plant-pathogenic bacteria and fungi. They act rapidly, at low micromolar concentrations, on the membranes of the microorganisms via a lytic mechanism. In vitro microscopic analysis revealed wide-scale damage to the microorganism's membrane, in addition to inhibition of pathogen growth. In planta potent antifungal activity was demonstrated on cucumber fruits and leaves infected with the pathogen Botrytis cinerea as well as on corn leaves infected with Cochliobolus heterostrophus. Similarly, treatment with the lipopeptides of Arabidopsis leaves infected with the bacterial leaf pathogen Pseudomonas syringae efficiently and rapidly reduced the number of bacteria. Importantly, in contrast to what occurred with many native lipopeptides, no toxicity was observed on the plant tissues. These data suggest that the ultrashort lipopeptides could serve as native-like antimicrobial agents economically feasible for use in plant protection.

  3. Dynamic root exudation of sorgoleone and its in planta mechanism of action

    PubMed Central

    Dayan, Franck E.; Howell, J'Lynn; Weidenhamer, Jeffrey D.

    2009-01-01

    The oily droplets exuded from the root hairs of sorghum are composed of a 1:1 ratio of sorgoleone and its lipid resorcinol analogue. The production of these droplets appears to be suppressed when c. 20 μg of exudate mg−1 root dry weight accumulates at the tip of the root hairs. However, more exudate is produced following gentle washing of the roots with water, suggesting that the biosynthesis of lipid benzoquinones and resorcinols is a dynamic process. Sorgoleone interferes with several molecular target sites, including photosynthetic electron transport, in in vitro assays. However, the in planta mechanism of action of sorgoleone remains controversial because it is not clear whether this lipid benzoquinone exuding from the roots of sorghum is taken up by roots of the receiving plants and translocated to their foliage where it must enter the chloroplast and inhibit PSII in the thylakoid membrane. Experiments designed to test the in planta mode of action of sorgoleone demonstrated that it has no effect on the photosynthesis of older plants, but inhibits photosynthesis in germinating seedlings. Sorgoleone is not translocated acropetally in older plants, but can be absorbed through the hypocotyl and cotyledonary tissues. Therefore, the mode of action of sorgoleone may be the result of inhibition of photosynthesis in young seedlings in concert with inhibition of its other molecular target sites in older plants. PMID:19357432

  4. Bcmimp1, a Botrytis cinerea Gene Transiently Expressed in planta, Encodes a Mitochondrial Protein

    PubMed Central

    Benito-Pescador, David; Santander, Daniela; Arranz, M.; Díaz-Mínguez, José M.; Eslava, Arturo P.; van Kan, Jan A. L.; Benito, Ernesto P.

    2016-01-01

    Botrytis cinerea is a widespread necrotrophic fungus which infects more than 200 plant species. In an attempt to characterize the physiological status of the fungus in planta and to identify genetic factors contributing to its ability to infect the host cells, a differential gene expression analysis during the interaction B. cinerea-tomato was carried out. Gene Bcmimp1 codes for a mRNA detected by differential display in the course of this analysis. During the interaction with the host, it shows a transient expression pattern with maximal expression levels during the colonization and maceration of the infected tissues. Bioinformatic analysis suggested that BCMIMP1 is an integral membrane protein located in the mitochondrial inner membrane. Co-localization experiments with a BCMIMP1-GFP fusion protein confirmed that the protein is targeted to the mitochondria. ΔBcmimp1 mutants do not show obvious phenotypic differences during saprophytic growth and their infection ability was unaltered as compared to the wild-type. Interestingly, the mutants produced increased levels of reactive oxygen species, likely as a consequence of disturbed mitochondrial function. Although Bcmimp1 expression is enhanced in planta it cannot be considered a pathogenicity factor. PMID:26952144

  5. Inhibition of Fungal and Bacterial Plant Pathogens In Vitro and In Planta with Ultrashort Cationic Lipopeptides▿

    PubMed Central

    Makovitzki, Arik; Viterbo, Ada; Brotman, Yariv; Chet, Ilan; Shai, Yechiel

    2007-01-01

    Plant diseases constitute an emerging threat to global food security. Many of the currently available antimicrobial agents for agriculture are highly toxic and nonbiodegradable and cause extended environmental pollution. Moreover, an increasing number of phytopathogens develop resistance to them. Recently, we have reported on a new family of ultrashort antimicrobial lipopeptides which are composed of only four amino acids linked to fatty acids (A. Makovitzki, D. Avrahami, and Y. Shai, Proc. Natl. Acad. Sci. USA 103:15997-16002, 2006). Here, we investigated the activities in vitro and in planta and the modes of action of these short lipopeptides against plant-pathogenic bacteria and fungi. They act rapidly, at low micromolar concentrations, on the membranes of the microorganisms via a lytic mechanism. In vitro microscopic analysis revealed wide-scale damage to the microorganism's membrane, in addition to inhibition of pathogen growth. In planta potent antifungal activity was demonstrated on cucumber fruits and leaves infected with the pathogen Botrytis cinerea as well as on corn leaves infected with Cochliobolus heterostrophus. Similarly, treatment with the lipopeptides of Arabidopsis leaves infected with the bacterial leaf pathogen Pseudomonas syringae efficiently and rapidly reduced the number of bacteria. Importantly, in contrast to what occurred with many native lipopeptides, no toxicity was observed on the plant tissues. These data suggest that the ultrashort lipopeptides could serve as native-like antimicrobial agents economically feasible for use in plant protection. PMID:17720828

  6. In planta expression of hyperthermophilic enzymes as a strategy for accelerated lignocellulosic digestion.

    PubMed

    Mir, Bilal Ahmad; Myburg, Alexander A; Mizrachi, Eshchar; Cowan, Don A

    2017-09-13

    Conversion of lignocellulosic biomass to biofuels and biomaterials suffers from high production costs associated with biomass pretreatment and enzymatic hydrolysis. In-planta expression of lignocellulose-digesting enzymes is a promising approach to reduce these cost elements. However, this approach faces a number of challenges, including auto-hydrolysis of developing cell walls, plant growth and yield penalties, low expression levels and the limited stability of expressed enzymes at the high temperatures generally used for biomass processing to release fermentable sugars. To overcome these challenges we expressed codon-optimized recombinant hyperthermophilic endoglucanase (EG) and xylanase (Xyn) genes in A. thaliana. Transgenic Arabidopsis lines expressing EG and Xyn enzymes at high levels without any obvious plant growth or yield penalties were selected for further analysis. The highest enzyme activities were observed in the dry stems of transgenic lines, indicating that the enzymes were not degraded during stem senescence and storage. Biomass from transgenic lines exhibited improved saccharification efficiency relative to WT control plants. We conclude that the expression of hyperthermophilic enzymes in plants is a promising approach for combining pretreatment and enzymatic hydrolysis processes in lignocellulosic digestion. This study provides a valid foundation for further studies involving in planta co-expression of core and accessory lignocellulose-digesting enzymes.

  7. Prediction of the in planta P. pachyrhizi secretome and potential effector families.

    PubMed

    De Carvalho, Mayra C da C G; Nascimento, Leandro Costa; Darben, Luana M; Polizel-Podanosqui, Adriana M; Lopes-Caitar, Valéria S; Qi, Mingsheng; Rocha, Carolina S; Carazzolle, Marcelo Falsarella; Kuwahara, Márcia K; Pereira, Goncalo A G; Abdelnoor, Ricardo V; Whitham, Steven A; Marcelino-Guimarães, Francismar C

    2016-03-24

    Asian soybean rust (ASR) caused by the obligate biotrophic fungus Phakopsora pachyrhizi can cause losses greater than 80%. Despite its economic importance, there is no soybean cultivar with durable ASR resistance (Goellner et al., 2010). In addition, the P. pachyrhizi genome is not yet available. However, the availability of other rust genomes as well as the development of sample enrichment strategies and bioinformatics tools has improved our knowledge of the ASR secretome and its potential effectors. In this context, we used a combination of laser capture microdissection (LCM), RNAseq and a bioinformatics pipeline to identify a total of 36,350 P. pachyrhizi contigs expressed in planta and a predicted secretome of 851 proteins. Some of the predicted secreted proteins had characteristics of candidate effectors, including small size, cysteine rich, do not contain PFAM domains (except those associated with pathogenicity), and were strongly expressed in planta. A comparative analysis of the predicted secreted proteins present in Pucciniales species identified new members of soybean rust and new Pucciniales- or P. pachyrhizi-specific families (tribes). Members of some families were strongly upregulated during early infection, starting with initial infection through haustorium formation. Effector candidates selected from two of these families were able to suppress immunity in transient assays, and were localized in the plant cytoplasm and nuclei. These experiments support our bioinformatics predictions and show that these families contain members that have functions consistent with P. pachyrhizi effectors. This article is protected by copyright. All rights reserved.

  8. Bcmimp1, a Botrytis cinerea Gene Transiently Expressed in planta, Encodes a Mitochondrial Protein.

    PubMed

    Benito-Pescador, David; Santander, Daniela; Arranz, M; Díaz-Mínguez, José M; Eslava, Arturo P; van Kan, Jan A L; Benito, Ernesto P

    2016-01-01

    Botrytis cinerea is a widespread necrotrophic fungus which infects more than 200 plant species. In an attempt to characterize the physiological status of the fungus in planta and to identify genetic factors contributing to its ability to infect the host cells, a differential gene expression analysis during the interaction B. cinerea-tomato was carried out. Gene Bcmimp1 codes for a mRNA detected by differential display in the course of this analysis. During the interaction with the host, it shows a transient expression pattern with maximal expression levels during the colonization and maceration of the infected tissues. Bioinformatic analysis suggested that BCMIMP1 is an integral membrane protein located in the mitochondrial inner membrane. Co-localization experiments with a BCMIMP1-GFP fusion protein confirmed that the protein is targeted to the mitochondria. ΔBcmimp1 mutants do not show obvious phenotypic differences during saprophytic growth and their infection ability was unaltered as compared to the wild-type. Interestingly, the mutants produced increased levels of reactive oxygen species, likely as a consequence of disturbed mitochondrial function. Although Bcmimp1 expression is enhanced in planta it cannot be considered a pathogenicity factor.

  9. Biosynthesis, structural, and functional attributes of tocopherols in planta; past, present, and future perspectives.

    PubMed

    Hussain, Nazim; Irshad, Faiza; Jabeen, Zahra; Shamsi, Imran Haider; Li, Zhilan; Jiang, Lixi

    2013-07-03

    Tocopherols are lipophilic molecules, ubiquitously synthesized in all photosynthetic organisms. Being a group of vitamin E compounds, they play an essential role in human nutrition and health. Despite their structural and functional attributes as important antioxidants in plants, it would be misleading to ignore the potential roles of tocopherols beyond their antioxidant properties in planta. Detailed characterization of mutants and transgenic plants, including Arabidopsis (vte1, vte2, vte4, and so on), maize (sxd1) mutants, and transgenic potato and tobacco lines altered in tocopherol biosynthesis and contents, has led to surprising outcomes regarding the additional functions of these molecules. Thus, the aim of this review is to highlight the past and present research findings on tocopherols' structural, biosynthesis, and functional properties in plants. Special emphasis is given to their suggested functions in planta, such as cell signaling, hormonal interactions, and coordinated response of tocopherols to other antioxidants under abiotic stresses. Moreover, some important questions about possible new functions of tocopherols will be discussed as future prospects to stimulate further research.

  10. Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH).

    PubMed

    Vági, Pál; Knapp, Dániel G; Kósa, Annamária; Seress, Diána; Horváth, Áron N; Kovács, Gábor M

    2014-05-01

    In planta detection of mutualistic, endophytic, and pathogenic fungi commonly colonizing roots and other plant organs is not a routine task. We aimed to use fluorescence in situ hybridization (FISH) for simultaneous specific detection of different fungi colonizing the same tissue. We have adapted ribosomal RNA (rRNA) FISH for visualization of common mycorrhizal (arbuscular- and ectomycorrhiza) and endophytic fungi within roots of different plant species. Beside general probes, we designed and used specific ones hybridizing to the large subunit of rRNA with fluorescent dyes chosen to avoid or reduce the interference with the autofluorescence of plant tissues. We report here an optimized efficient protocol of rRNA FISH and the use of both epifluorescence and confocal laser scanning microscopy for simultaneous specific differential detection of those fungi colonizing the same root. The method could be applied for the characterization of other plant-fungal interactions, too. In planta FISH with specific probes labeled with appropriate fluorescent dyes could be used not only in basic research but to detect plant colonizing pathogenic fungi in their latent life-period.

  11. Comparative analysis of the in vitro and in planta secretomes from Mycosphaerella fijiensis isolates.

    PubMed

    Escobar-Tovar, Lina; Guzmán-Quesada, Mauricio; Sandoval-Fernández, Jorge A; Gómez-Lim, Miguel A

    2015-06-01

    Black Sigatoka, a devastating disease of bananas and plantains worldwide, is caused by the fungus Mycosphaerella fijiensis. Several banana cultivars such as 'Yangambi Km 5' and Calcutta IV, have been known to be resistant to the fungus, but the resistance has been broken in 'Yangambi Km 5' in Costa Rica. Since the resistance of this variety still persists in Mexico, the aim of this study was to compare the in vitro and in planta secretomes from two avirulent and virulent M. fijiensis isolates using proteomics and bioinformatics approaches. We aimed to identify differentially expressed proteins in fungal isolates that differ in pathogenicity and that might be responsible for breaking the resistance in 'Yangambi Km 5'. We were able to identify 90 protein spots in the secretomes of fungal isolates encoding 42 unique proteins and 35 differential spots between them. Proteins involved in carbohydrate transport and metabolism were more prevalent. Several proteases, pathogenicity-related, ROS detoxification and unknown proteins were also highly or specifically expressed by the virulent isolate in vitro or during in planta infection. An unknown protein representing a virulence factor candidate was also identified. These results demonstrated that the secretome reflects major differences between both M. fijiensis isolates. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  12. In-planta Sporulation Capacity Enhances Infectivity and Rhizospheric Competitiveness of Frankia Strains

    PubMed Central

    Cotin-Galvan, Laetitia; Pozzi, Adrien C.; Schwob, Guillaume; Fournier, Pascale; Fernandez, Maria P.; Herrera-Belaroussi, Aude

    2016-01-01

    Frankia Sp+ strains maintain their ability to sporulate in symbiosis with actinorhizal plants, producing abundant sporangia inside host plant cells, in contrast to Sp− strains, which are unable to perform in-planta sporulation. We herein examined the role of in-planta sporulation in Frankia infectivity and competitiveness for root infection. Fifteen strains belonging to different Sp+ and Sp− phylogenetic lineages were inoculated on seedlings of Alnus glutinosa (Ag) and A. incana (Ai). Strain competitiveness was investigated by performing Sp−/Sp+ co-inoculations. Plant inoculations were standardized using crushed nodules obtained under laboratory-controlled conditions (same plant species, age, and environmental factors). Specific oligonucleotide primers were developed to identify Frankia Sp+ and/or Sp− strains in the resulting nodules. Single inoculation experiments showed that (i) infectivity by Sp+ strains was significantly greater than that by Sp− strains, (ii) genetically divergent Sp+ strains exhibited different infective abilities, and (iii) Sp+ and Sp− strains showed different host preferences according to the origin (host species) of the inocula. Co-inoculations of Sp+ and Sp− strains revealed the greater competitiveness of Sp+ strains (98.3 to 100% of Sp+ nodules, with up to 15.6% nodules containing both Sp+ and Sp− strains). The results of the present study highlight differences in Sp+/Sp− strain ecological behaviors and provide new insights to strengthen the obligate symbiont hypothesis for Sp+ strains. PMID:26726131

  13. In planta production and characterization of a hyperthermostable GH10 xylanase in transgenic sugarcane.

    PubMed

    Kim, Jae Yoon; Nong, Guang; Rice, John D; Gallo, Maria; Preston, James F; Altpeter, Fredy

    2017-03-01

    Sugarcane (Saccharum sp. hybrids) is one of the most efficient and sustainable feedstocks for commercial production of fuel ethanol. Recent efforts focus on the integration of first and second generation bioethanol conversion technologies for sugarcane to increase biofuel yields. This integrated process will utilize both the cell wall bound sugars of the abundant lignocellulosic sugarcane residues in addition to the sucrose from stem internodes. Enzymatic hydrolysis of lignocellulosic biomass into its component sugars requires significant amounts of cell wall degrading enzymes. In planta production of xylanases has the potential to reduce costs associated with enzymatic hydrolysis but has been reported to compromise plant growth and development. To address this problem, we expressed a hyperthermostable GH10 xylanase, xyl10B in transgenic sugarcane which displays optimal catalytic activity at 105 °C and only residual catalytic activity at temperatures below 70 °C. Transgene integration and expression in sugarcane were confirmed by Southern blot, RT-PCR, ELISA and western blot following biolistic co-transfer of minimal expression cassettes of xyl10B and the selectable neomycin phosphotransferase II. Xylanase activity was detected in 17 transgenic lines with a fluorogenic xylanase activity assay. Up to 1.2% of the total soluble protein fraction of vegetative progenies with integration of chloroplast targeted expression represented the recombinant Xyl10B protein. Xyl10B activity was stable in vegetative progenies. Tissues retained 75% of the xylanase activity after drying of leaves at 35 °C and a 2 month storage period. Transgenic sugarcane plants producing Xyl10B did not differ from non-transgenic sugarcane in growth and development under greenhouse conditions. Sugarcane xylan and bagasse were used as substrate for enzymatic hydrolysis with the in planta produced Xyl10B. TLC and HPLC analysis of hydrolysis products confirmed the superior catalytic activity

  14. Mexican papita viroid and tomato planta macho viroid belong to a single species in the genus Pospiviroid

    USDA-ARS?s Scientific Manuscript database

    Tomato planta macho viroid (TPMVd) and Mexican papita viroid (MPVd) are two closely-related (>90% sequence identity) members of the genus Pospiviroid. Their current status as separate species is based upon the reported ability of TPMVd to replicate in Gomphrena globosa and the inability of this viro...

  15. Single, Ancient Origin of a Plastid Metabolite Translocator Family in Plantae from an Endomembrane-Derived Ancestor

    PubMed Central

    Weber, Andreas P. M.; Linka, Marc; Bhattacharya, Debashish

    2006-01-01

    Phylogenetic analyses show the single origin of a plastid metabolite translocator family in the Plantae from a gene encoding an existing endomembrane-derived protein. Red algal secondary endosymbiosis has spread a translocator gene into the ancestor of the “chromalveolate” protists, where it has diversified into a novel clade of proteins. PMID:16524915

  16. The Ustilago maydis b mating type locus controls hyphal proliferation and expression of secreted virulence factors in planta.

    PubMed

    Wahl, Ramon; Zahiri, Alexander; Kämper, Jörg

    2010-01-01

    Sexual development in fungi is controlled by mating type loci that prevent self-fertilization. In the phytopathogenic fungus Ustilago maydis, the b mating type locus encodes two homeodomain proteins, termed bE and bW. After cell fusion, a heterodimeric bE/bW complex is formed if the proteins are derived from different alleles. The bE/bW complex is required and sufficient to initiate pathogenic development and sexual reproduction; for the stages of pathogenic development succeeding plant penetration, however, its role was unclear. To analyse b function during in planta development, we generated a temperature-sensitive bE(ts) protein by exchange of a single amino acid. bE(ts) strains are stalled in pathogenic development at restrictive temperature in planta, and hyphae develop enlarged, bulbous cells at their tips that contain multiple nuclei, indicating a severe defect in the control and synchronization of cell cycle and cytokinesis. DNA array analysis of bE(ts) mutant strains in planta revealed a b-dependent regulation of genes encoding secreted proteins that were shown to influence fungal virulence. Our data demonstrate that in U. maydis the b heterodimer is not only essential to establish the heterodikaryon after mating of two compatible sporidia and to initiate fungal pathogenicity, but also to sustain in planta proliferation and ensure sexual reproduction.

  17. Microbial secondary metabolites ameliorate growth, in planta contents and lignification in Withania somnifera (L.) Dunal.

    PubMed

    Singh, Akanksha; Gupta, Rupali; Srivastava, Madhumita; Gupta, M M; Pandey, Rakesh

    2016-04-01

    In the present investigation, metabolites of Streptomyces sp. MTN14 and Trichoderma harzianum ThU significantly enhanced biomass yield (3.58 and 3.48 fold respectively) in comparison to the control plants. The secondary metabolites treatments also showed significant augmentation (0.75-2.25 fold) in withanolide A, a plant secondary metabolite. Lignin deposition, total phenolic and flavonoid content in W. somnifera were maximally induced in treatment having T. harzianum metabolites. Also, Trichoderma and Streptomyces metabolites were found much better in invoking in planta contents and antioxidants compared with their live culture treatments. Therefore, identification of new molecular effectors from metabolites of efficient microbes may be used as biopesticide and biofertilizer for commercial production of W. somnifera globally.

  18. In Planta Response of Arabidopsis to Photothermal Impact Mediated by Gold Nanoparticles.

    PubMed

    Koo, Yeonjong; Lukianova-Hleb, Ekaterina Y; Pan, Joann; Thompson, Sean M; Lapotko, Dmitri O; Braam, Janet

    2016-02-03

    Biological responses to photothermal effects of gold nanoparticles (GNPs) have been demonstrated and employed for various applications in diverse systems except for one important class - plants. Here, the uptake of GNPs through Arabidopsis thaliana roots and translocation to leaves are reported. Successful plasmonic nanobubble generation and acoustic signal detection in planta is demonstrated. Furthermore, Arabidopsis leaves harboring GNPs and exposed to continuous laser or noncoherent light show elevated temperatures across the leaf surface and induced expression of heat-shock regulated genes. Overall, these results demonstrate that Arabidopsis can readily take up GNPs through the roots and translocate the particles to leaf tissues. Once within leaves, GNPs can act as photothermal agents for on-demand remote activation of localized biological processes in plants. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. In Planta Microsphere-Based Lateral Flow Leaf Biosensor in Maize.

    PubMed

    Wen, Jessica T; Castro, Carlos; Tsutsui, Hideaki

    2015-08-01

    Low-cost and quick detection of biotic stresses is critically important for protection of staple food crops such as maize in smallholder farms in developing countries, where access to improved seed varieties, fertilizers, and pesticides is limited due to financial and geographical reasons. Here, we report a new lateral flow detection technology directly integrated in a maize leaf, in which microspheres conjugated with analyte-specific capture antibodies are non-invasively injected. The antibody-conjugated microspheres capture and detect an analyte in a concentration-specific manner. In this study, we optimized microsphere size for effective infiltration and immobilization in the leaf, and further demonstrated detection of a fluorescent mock biomarker, fluorescein, in a live maize plant. This in planta lateral flow biosensor is the first of its kind and is expected to provide a low-cost and user-friendly detection method for biotic stresses in the field. © 2014 Society for Laboratory Automation and Screening.

  20. In Planta Localization of Stilbenes within Picea abies Phloem1[OPEN

    PubMed Central

    Kuroda, Katsushi; Pranovich, Andrey; Roig-Juan, Sílvia; Aoki, Dan

    2016-01-01

    Phenolic stilbene glucosides (astringin, isorhapontin, and piceid) and their aglycons commonly accumulate in the phloem of Norway spruce (Picea abies). However, current knowledge about the localization and accumulation of stilbenes within plant tissues and cells remains limited. Here, we used an innovative combination of novel microanalytical techniques to evaluate stilbenes in a frozen-hydrated condition (i.e. in planta) and a freeze-dried condition across phloem tissues. Semiquantitative time-of-flight secondary ion-mass spectrometry imaging in planta revealed that stilbenes were localized in axial parenchyma cells. Quantitative gas chromatography analysis showed the highest stilbene content in the middle of collapsed phloem with decreases toward the outer phloem. The same trend was detected for soluble sugar and water contents. The specimen water content may affect stilbene composition; the glucoside-to-aglycon ratio decreased slightly with decreases in water content. Phloem chemistry was correlated with three-dimensional structures of phloem as analyzed by microtomography. The outer phloem was characterized by a high volume of empty parenchyma, reduced ray volume, and a large number of axial parenchyma with porous vacuolar contents. Increasing porosity from the inner to the outer phloem was related to decreasing compactness of stilbenes and possible secondary oxidation or polymerization. Our results indicate that aging-dependent changes in phloem may reduce cell functioning, which affects the capacity of the phloem to store water and sugar, and may reduce the defense potential of stilbenes in the axial parenchyma. Our results highlight the power of using a combination of techniques to evaluate tissue- and cell-level mechanisms involved in plant secondary metabolite formation and metabolism. PMID:27531441

  1. In planta transformation method for T-DNA transfer in orchids

    NASA Astrophysics Data System (ADS)

    Semiarti, Endang; Purwantoro, Aziz; Mercuriani, Ixora S.; Anggriasari, Anida M.; Jang, Seonghoe; Suhandono, Sony; Machida, Yasunori; Machida, Chiyoko

    2014-03-01

    Transgenic plant technology is an efficient tool to study the function of gene(s) in plant. The most popular and widely used technique is Agrobacterium-mediated transformation in which cocultivation was done by immersing the plant tissues/organ in overnight bacterial cultured for about 30 minutes to one hour under in vitro condition. In this experiment, we developed more easier technique that omitted the in vitro step during cocultivation with Agrobacterium, namely in planta transformation method. Pollinaria (compact pollen mass of orchid) of Phalaenopsis amabilis and Spathoglottis plicata orchids were used as target explants that were immersed into bacterial culture for 30 minutes, then dried up the pollinaria, the transformed pollinaria was used to pollinate orchid flowers. The T-DNA used for this experiments were Ubipro∷PaFT/A. tumefaciens GV3101 for P. amabilis and MeEF1α2 pro∷GUS/ A. tumefaciens LBA 4404 for S.plicata. Seeds that were produced from pollinated flowers were grown onto 10 mg/l hygromicin containing NP (New Phalaenopsis) medium. The existance of transgene in putative transformant protocorm (developing orchid embryo) genome was confirmed using PCR with specific primers of either PaFT or GUS genes. Histochemical GUS assay was also performed to the putative transformants. The result showed that transformation frequencies were 2.1 % in P. amabilis, and 0,53% in S. plicata. These results indicates that in planta transformation method could be used for Agrobacterium-mediated genetic transformation, with advantage easier and more secure work from contaminants than that of the in vitro method.

  2. Large-scale gene discovery in the septoria tritici blotch fungus Mycosphaerella graminicola with a focus on in planta expression.

    PubMed

    Kema, Gert H J; van der Lee, Theo A J; Mendes, Odette; Verstappen, Els C P; Lankhorst, René Klein; Sandbrink, Hans; van der Burgt, Ate; Zwiers, Lute-Harm; Csukai, Michael; Waalwijk, Cees

    2008-09-01

    The foliar disease septoria tritici blotch, caused by the fungus Mycosphaerella graminicola, is currently the most important wheat disease in Europe. Gene expression was examined under highly different conditions, using 10 expressed sequence tag libraries generated from M. graminicola isolate IPO323 using seven in vitro and three in planta growth conditions. To identify fungal clones in the interaction libraries, we developed a selection method based on hybridization with the entire genomic DNA of M. graminicola, to selectively enrich these libraries for fungal genes. Assembly of the 27,007 expressed sequence tags resulted in 9,190 unigenes, representing 5.2 Mb of the estimated 39-Mb genome size of M. graminicola. All libraries contributed significantly to the number of unigenes, especially the in planta libraries representing different stages of pathogenesis, which covered 15% of the library-specific unigenes. Even under presymptomatic conditions (5 days postinoculation), when fungal biomass is less than 5%, this method enabled us to efficiently capture fungal genes expressed during pathogenesis. Many of these genes were uniquely expressed in planta, indicating that in planta gene expression significantly differed from in vitro expression. Examples of gene discovery included a number of cell wall-degrading enzymes, a broad set of genes involved in signal transduction (n=11) and a range of ATP-binding cassette (n=20) and major facilitator superfamily transporter genes (n=12) potentially involved in protection against antifungal compounds or the secretion of pathogenicity factors. In addition, evidence is provided for a mycovirus in M. graminicola that is highly expressed under various stress conditions, in particular, under nitrogen starvation. Our analyses provide a unique window on in vitro and in planta gene expression of M. graminicola.

  3. XerR, a negative regulator of XccR in Xanthomonas campestris pv. campestris, relieves its repressor function in planta.

    PubMed

    Wang, Li; Zhang, Lili; Geng, Yunfeng; Xi, Wei; Fang, Rongxiang; Jia, Yantao

    2011-07-01

    We previously reported that XccR, a LuxR-type regulator of Xanthomonas campestris pv. campestris (Xcc), activates the downstream proline iminopeptidase virulence gene (pip) in response to certain host plant factor(s). In this report, we further show that the expression of the xccR gene was repressed in the culture medium by an NtrC-type response regulator, which we named XerR (XccR expression-related, repressor), and that this repression was relieved when the bacteria were grown in planta. Such a regulatory mechanism is reinforced by the observations that XerR directly bound to the xccR promoter in vitro, and that mutations at the phosphorylation-related residues of XerR resulted in the loss of its repressor function. Furthermore, the expression level of xccR increased even in XerR-overexpressing Xcc cells when they were vacuum infiltrated into cabbage plants. We also preliminarily characterized the host factor(s) involved in the above mentioned interactions between Xcc and the host plant, showing that a plant material(s) with molecular weight(s) less than 1 kDa abolished the binding of XerR to the xccR promoter, while the same material enhanced the binding of XccR to the luxXc box in the pip promoter. Taken together, our results implicate XerR in a new layer of the regulatory mechanism controlling the expression of the virulence-related xccR/pip locus and provide clues to the identification of plant signal molecules that interact with XerR and XccR to enhance the virulence of Xcc.

  4. XerR, a negative regulator of XccR in Xanthomonas campestris pv. campestris, relieves its repressor function in planta

    PubMed Central

    Wang, Li; Zhang, Lili; Geng, Yunfeng; Xi, Wei; Fang, Rongxiang; Jia, Yantao

    2011-01-01

    We previously reported that XccR, a LuxR-type regulator of Xanthomonas campestris pv. campestris (Xcc), activates the downstream proline iminopeptidase virulence gene (pip) in response to certain host plant factor(s). In this report, we further show that the expression of the xccR gene was repressed in the culture medium by an NtrC-type response regulator, which we named XerR (XccR expression-related, repressor), and that this repression was relieved when the bacteria were grown in planta. Such a regulatory mechanism is reinforced by the observations that XerR directly bound to the xccR promoter in vitro, and that mutations at the phosphorylation-related residues of XerR resulted in the loss of its repressor function. Furthermore, the expression level of xccR increased even in XerR-overexpressing Xcc cells when they were vacuum infiltrated into cabbage plants. We also preliminarily characterized the host factor(s) involved in the above mentioned interactions between Xcc and the host plant, showing that a plant material(s) with molecular weight(s) less than 1 kDa abolished the binding of XerR to the xccR promoter, while the same material enhanced the binding of XccR to the luxXc box in the pip promoter. Taken together, our results implicate XerR in a new layer of the regulatory mechanism controlling the expression of the virulence-related xccR/pip locus and provide clues to the identification of plant signal molecules that interact with XerR and XccR to enhance the virulence of Xcc. PMID:21483448

  5. Pea lectin receptor-like kinase promotes high salinity stress tolerance in bacteria and expresses in response to stress in planta.

    PubMed

    Joshi, Amita; Dang, Hung Quang; Vaid, Neha; Tuteja, Narendra

    2010-01-01

    The plant lectin receptor-like kinases (LecRLKs) are involved in various signaling pathways but their role in salinity stress tolerance has not heretofore been well described. Salinity stress negatively affects plant growth/productivity and threatens food security worldwide. Based on functional gene-mining assay, we have isolated 34 salinity tolerant genes out of one million Escherichia coli (SOLR) transformants containing pea cDNAs grown in 0.8 M NaCl. Sequence analysis of one of these revealed homology to LecRLK, which possesses N-myristilation and N-glycosylation sites thus corroborating the protein to be a glycoconjugate. The homology based computational modeling of the kinase domain suggested high degree of conservation with the protein already known to be stress responsive in plants. The NaCl tolerance provided by PsLecRLK to the above bacteria was further confirmed in E. coli (DH5alpha). In planta studies showed that the expression of PsLecRLK cDNA was significantly upregulated in response to NaCl as compared to K(+) and Li(+) ions, suggesting the Na(+) ion specific response. Transcript of the PsLecRLK gene accumulates mainly in roots and shoots. The purified 47 kDa recombinant PsLecRLK-KD (kinase domain) protein has been shown to phosphorylate general substrates like MBP and casein. This study not only suggests the conservation of the cellular response to high salinity stress across prokaryotes and plant kingdom but also provides impetus to develop novel concepts for better understanding of mechanism of stress tolerance in bacteria and plants. It also opens up new avenues for studying practical aspects of plant salinity tolerance for enhanced agricultural productivity.

  6. Towards an Understanding of Enhanced Biomass Digestibility by In Planta Expression of a Family 5 Glycoside Hydrolase.

    PubMed

    Donohoe, Bryon S; Wei, Hui; Mittal, Ashutosh; Shollenberger, Todd; Lunin, Vladimir V; Himmel, Michael E; Brunecky, Roman

    2017-06-29

    In planta expression of a thermophilic endoglucanase (AcCel5A) reduces recalcitrance by creating voids and other irregularities in cell walls of Arabidopsis thaliana that increase enzyme accessibility without negative impacts on plant growth or cell wall composition. Our results suggest that cellulose β-1-4 linkages can be cut sparingly in the assembling wall and that these minimal changes, made at the proper time, have an impact on plant cell wall recalcitrance without negative effects on overall plant development.

  7. Root internalization, transport and in-planta survival of Salmonella enterica serovar Newport in sweet basil.

    PubMed

    Gorbatsevich, Elena; Sela Saldinger, Shlomo; Pinto, Riky; Bernstein, Nirit

    2013-02-01

    It is now acknowledged that food-borne pathogens present in the irrigation water or soil can become associated with crop plants in the field, penetrate internal plant tissues via the root, translocate and survive inside plants. Only little information is available concerning interaction between enteric pathogens and plants. The present study evaluated the potential for contamination of the aromatic plant, sweet basil during cultivation, by Salmonella enterica serovar Newport. Root internalization was plant-age-dependent, with the highest susceptibility occurring at the beginning of the rapid growth phase of the root. Higher incidence of internalization was detected in vegetative than reproductive plant organs, pointing at bacterial transport in the transpiration stream. Internalized Salmonella survived only < 30 h in the phyllosphere. In contrast, survival of Salmonella on the leaf surface was much pronounced (at least 8 days), and the initial decay rate was lower at the abaxial (lower) compared with the adaxial (upper) side of the leaf. Although the experiments were conducted with high concentration of Salmonella unlikely to happen in the field, internalization occurred at a low frequency and in-planta survival was limited to less than 30 h. These findings imply that leaf surface contamination, rather than root internalization, may pose higher risk for human infection following consumption of contaminated basil. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  8. In planta production of flock house virus transencapsidated RNA and its potential use as a vaccine.

    PubMed

    Zhou, Yiyang; Maharaj, Payal D; Mallajosyula, Jyothi K; McCormick, Alison A; Kearney, Christopher M

    2015-04-01

    We have developed a transencapsidated vaccine delivery system based on the insect virus, Flock House virus (FHV). FHV is attractive due to its small genome size, simple organization, and nonpathogenic characteristics. With the insertion of a Tobacco mosaic virus (TMV) origin of assembly (Oa), the independently replicating FHV RNA1 can be transencapsidated by TMV coat protein. In this study, we demonstrated that the Oa-adapted FHV RNA1 transencapsidation process can take place in planta, by using a bipartite plant expression vector system, where TMV coat protein is expressed by another plant virus vector, Foxtail mosaic virus (FoMV). Dual infection in the same cell by both FHV and FoMV was observed. Though an apparent classical coat protein-mediated resistance repressed FHV expression, this was overcome by delaying inoculation of the TMV coat protein vector by 3 days after FHV vector inoculation. Expression of the transgene marker in animals by these in vivo-generated transencapsidated nanoparticles was confirmed by mouse vaccination, which also showed an improved vaccine response compared to similar in vitro-produced vaccines.

  9. TYLCV-Is movement in planta does not require V2 protein

    SciTech Connect

    Hak, Hagit; Levy, Yael; Chandran, Sam A.; Belausov, Eduard; Loyter, Abraham; Lapidot, Moshe; Gafni, Yedidya

    2015-03-15

    Tomato yellow leaf curl virus (TYLCV), a major tomato pathogen causing extensive crop losses, is a whitefly-transmitted geminivirus. V2 mutants of TYLCV-Is and related viruses tend to induce symptomless infection with attenuated viral DNA levels, while accumulating close to wild-type DNA levels in protoplasts, suggesting V2 as a movement protein. The discovery of plant-silencing mechanisms and viral silencing suppressors, V2 included, led us to reconsider V2's involvement in viral movement. We studied two mutant versions of the virus, one impaired in V2 silencing-suppression activity, and another carrying a non-translatable V2. While both mutant viruses spread in the infected plant to newly emerged leaves at the same rate as the wild-type virus, their DNA-accumulation levels were tenfold lower than in the wild-type virus. Thus, we suggest that the setback in virus proliferation, previously ascribed to a movement impediment, is due to lack of silencing-suppression activity. - Highlights: • TYLCV-Is V2 protein is localized in distinct microbodies throughout the cell cytoplasm, around the nucleus and in association with cytoplasmic strands but is not associated with the plasmodesmata. • Disruption of RNA-silencing suppression activity of TYLCV-Is V2 protein causes low titer of the virus in the infected plants. • The movement of TYLCV-Is in planta does not require a functional V2 protein.

  10. In Planta Determination of the mRNA-Binding Proteome of Arabidopsis Etiolated Seedlings.

    PubMed

    Reichel, Marlene; Liao, Yalin; Rettel, Mandy; Ragan, Chikako; Evers, Maurits; Alleaume, Anne-Marie; Horos, Rastislav; Hentze, Matthias W; Preiss, Thomas; Millar, Anthony A

    2016-10-01

    RNA binding proteins (RBPs) control the fate and expression of a transcriptome. Despite this fundamental importance, our understanding of plant RBPs is rudimentary, being mainly derived via bioinformatic extrapolation from other kingdoms. Here, we adapted the mRNA-protein interactome capture method to investigate the RNA binding proteome in planta. From Arabidopsis thaliana etiolated seedlings, we captured more than 700 proteins, including 300 with high confidence that we have defined as the At-RBP set. Approximately 75% of these At-RBPs are bioinformatically linked with RNA biology, containing a diversity of canonical RNA binding domains (RBDs). As no prior experimental RNA binding evidence exists for the majority of these proteins, their capture now authenticates them as RBPs. Moreover, we identified protein families harboring emerging and potentially novel RBDs, including WHIRLY, LIM, ALBA, DUF1296, and YTH domain-containing proteins, the latter being homologous to animal RNA methylation readers. Other At-RBP set proteins include major signaling proteins, cytoskeleton-associated proteins, membrane transporters, and enzymes, suggesting the scope and function of RNA-protein interactions within a plant cell is much broader than previously appreciated. Therefore, our foundation data set has provided an unbiased insight into the RNA binding proteome of plants, on which future investigations into plant RBPs can be based.

  11. Endophytic Colonization and In Planta Nitrogen Fixation by a Herbaspirillum sp. Isolated from Wild Rice Species

    PubMed Central

    Elbeltagy, Adel; Nishioka, Kiyo; Sato, Tadashi; Suzuki, Hisa; Ye, Bin; Hamada, Toru; Isawa, Tsuyoshi; Mitsui, Hisayuki; Minamisawa, Kiwamu

    2001-01-01

    Nitrogen-fixing bacteria were isolated from the stems of wild and cultivated rice on a modified Rennie medium. Based on 16S ribosomal DNA (rDNA) sequences, the diazotrophic isolates were phylogenetically close to four genera: Herbaspirillum, Ideonella, Enterobacter, and Azospirillum. Phenotypic properties and signature sequences of 16S rDNA indicated that three isolates (B65, B501, and B512) belong to the Herbaspirillum genus. To examine whether Herbaspirillum sp. strain B501 isolated from wild rice, Oryza officinalis, endophytically colonizes rice plants, the gfp gene encoding green fluorescent protein (GFP) was introduced into the bacteria. Observations by fluorescence stereomicroscopy showed that the GFP-tagged bacteria colonized shoots and seeds of aseptically grown seedlings of the original wild rice after inoculation of the seeds. Conversely, for cultivated rice Oryza sativa, no GFP fluorescence was observed for shoots and only weak signals were observed for seeds. Observations by fluorescence and electron microscopy revealed that Herbaspirillum sp. strain B501 colonized mainly intercellular spaces in the leaves of wild rice. Colony counts of surface-sterilized rice seedlings inoculated with the GFP-tagged bacteria indicated significantly more bacterial populations inside the original wild rice than in cultivated rice varieties. Moreover, after bacterial inoculation, in planta nitrogen fixation in young seedlings of wild rice, O. officinalis, was detected by the acetylene reduction and 15N2 gas incorporation assays. Therefore, we conclude that Herbaspirillum sp. strain B501 is a diazotrophic endophyte compatible with wild rice, particularly O. officinalis. PMID:11679357

  12. Advances in alfalfa mosaic virus-mediated expression of anthrax antigen in planta

    SciTech Connect

    Brodzik, R.; Bandurska, K.; Deka, D.; Golovkin, M.; Koprowski, H. . E-mail: h_koprowski@jefferson.edu

    2005-12-16

    Plant viruses show great potential for production of pharmaceuticals in plants. Such viruses can harbor a small antigenic peptide(s) as a part of their coat proteins (CP) and elicit an antigen-specific immune response. Here, we report the high yield and consistency in production of recombinant alfalfa mosaic virus (AlMV) particles for specific presentation of the small loop 15 amino acid epitope from domain-4 of the Bacillus anthracis protective antigen (PA-D4s). The epitope was inserted immediately after the first 25 N-terminal amino acids of AlMV CP to retain genome activation and binding of CP to viral RNAs. Recombinant AlMV particles were efficiently produced in tobacco, easily purified for immunological analysis, and exhibited extended stability and systemic proliferation in planta. Intraperitional injections of mice with recombinant plant virus particles harboring the PA-D4s epitope elicited a distinct immune response. Western blotting and ELISA analysis showed that sera from immunized mice recognized both native PA antigen and the AlMV CP.

  13. Development of 4-methoxy-7-nitroindolinyl (MNI)-caged auxins which are extremely stable in planta.

    PubMed

    Hayashi, Ken-Ichiro; Kusaka, Naoyuki; Yamasaki, Soma; Zhao, Yunde; Nozaki, Hiroshi

    2015-10-15

    Phytohormone auxin is a master regulator in plant growth and development. Regulation of cellular auxin level plays a central role in plant development. Auxin polar transport system modulates an auxin gradient that determines plant developmental process in response to environmental conditions and developmental programs. Photolabile caged auxins allow optical control of artificial auxin gradients at cellular resolution. Especially, two-photon uncaging system achieves high spatiotemporal control of photolysis reaction at two-photon cross-section. However, the development of caged versions of auxin has been limited by the instability of the caged auxins to higher plant metabolic activities. Here, we describe the synthesis and application of highly stable caged auxins, 4-methoxy-7-nitroindolinyl (MNI)-caged auxins. Natural auxin, indole 3-acetic acid, and two synthetic auxins, 1-NAA and 2,4-D were caged by MNI caging group. MNI-caged auxins showed a high stability in planta and a rapid release the original auxin when photolyzed. We demonstrated that optical control of auxin-responsive gene expression and auxin-related physiological responses by using MNI-caged auxins. We anticipate that MNI-caged auxins will be an effective tool for high-resolution control of endogenous auxin level. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Cytorhabdovirus P protein suppresses RISC-mediated cleavage and RNA silencing amplification in planta.

    PubMed

    Mann, Krin S; Johnson, Karyn N; Carroll, Bernard J; Dietzgen, Ralf G

    2016-03-01

    Plant viruses have evolved to undermine the RNA silencing pathway by expressing suppressor protein(s) that interfere with one or more key components of this antiviral defense. Here we show that the recently identified RNA silencing suppressor (RSS) of lettuce necrotic yellows virus (LNYV), phosphoprotein P, binds to RNA silencing machinery proteins AGO1, AGO2, AGO4, RDR6 and SGS3 in protein-protein interaction assays when transiently expressed. In planta, we demonstrate that LNYV P inhibits miRNA-guided AGO1 cleavage and translational repression, and RDR6/SGS3-dependent amplification of silencing. Analysis of LNYV P deletion mutants identified a C-terminal protein domain essential for both local RNA silencing suppression and interaction with AGO1, AGO2, AGO4, RDR6 and SGS3. In contrast to other viral RSS known to disrupt AGO activity, LNYV P sequence does not contain any recognizable GW/WG or F-box motifs. This suggests that LNYV P may represent a new class of AGO binding proteins. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Distribution of Tomato planta macho viroid in germinating pollen and transmitting tract.

    PubMed

    Matsushita, Yosuke; Yanagisawa, Hironobu

    2017-09-23

    Vertical and horizontal pollen transmission is important for efficient infection by viroids. Vertical pollen transmission of viroids is attributed to the infection by viroid in the embryo sac through infected pollen. To identify the viroid infection in pollen and pollen tubes elongating through the transmitting tract, we used in situ hybridization to histochemically analyze the distribution of Tomato planta macho viroid (TPMVd) in pollen grains, the stigma, and style of petunia plants. TPMVd was present in the generative nucleus and vegetative nucleus of mature infected pollen grains and germinating pollen grains. During pollen tube growth, TPMVd was present in the vegetative nucleus and two sperm nuclei, which were generated by division of the generative nucleus in the style transmitting tract. These findings indicated that viroid infection in sperm nuclei is responsible for vertical pollen transmission of viroids. TPMVd infection from TPMVd-infected pollen tubes to the transmitting tract was not observed. In addition, TPMVd signals were not confirmed in the stigma and transmitting tract of TPMVd-infected petunia plants, suggesting that viroids may not replicate in these tissues at the stage of mature style. Therefore, TPMVd may leak from the pollen tube somewhere in the ovary, except in the transmitting tract, during the horizontal transmission of TPMVd.

  16. Advances in alfalfa mosaic virus-mediated expression of anthrax antigen in planta.

    PubMed

    Brodzik, R; Bandurska, K; Deka, D; Golovkin, M; Koprowski, H

    2005-12-16

    Plant viruses show great potential for production of pharmaceuticals in plants. Such viruses can harbor a small antigenic peptide(s) as a part of their coat proteins (CP) and elicit an antigen-specific immune response. Here, we report the high yield and consistency in production of recombinant alfalfa mosaic virus (AlMV) particles for specific presentation of the small loop 15 amino acid epitope from domain-4 of the Bacillus anthracis protective antigen (PA-D4s). The epitope was inserted immediately after the first 25 N-terminal amino acids of AlMV CP to retain genome activation and binding of CP to viral RNAs. Recombinant AlMV particles were efficiently produced in tobacco, easily purified for immunological analysis, and exhibited extended stability and systemic proliferation in planta. Intraperitional injections of mice with recombinant plant virus particles harboring the PA-D4s epitope elicited a distinct immune response. Western blotting and ELISA analysis showed that sera from immunized mice recognized both native PA antigen and the AlMV CP.

  17. The signal transduction pathways controlling in planta tuberization in potato: an emerging synthesis.

    PubMed

    Sarkar, Debabrata

    2008-01-01

    Tuberization is one of the multiple outputs of a single-input phytochrome B sensory system, involving several regulatory genes. Phytochrome B- and GA-mediated photoperiodic perception occurs in the leaf, and then the RNA acts as a systemic signal in the long-distance signaling pathway to initiate tuberization in the subapical region of an underground stolon. There is good evidence that flowering and tuberizing signals might be similar. Is there a cross-talk with an oxidative burst-mediated redox signaling pathway during tuberization? Is the lipoxygenase cascade involved in the formation of the perimedullary tissue in a growing tuber? Do aquaporins regulate cell division, expansion and elongation during stolon growth and tuber induction in potato? Is the adaptive diversity for tuberization under varying photoperiods a micro-evolutionary indicator of differential transduction of cell-to-cell signal molecules under spatial and temporal expression of regulatory genes encoding transcriptional activators? Taking these views into consideration, the review presents an interim synthesis of a signaling network regulating in planta tuberization in potato.

  18. Assessing environmental impacts of genetically modified plants on non-target organisms: The relevance of in planta studies.

    PubMed

    Arpaia, Salvatore; Birch, A Nicholas E; Kiss, Jozsef; van Loon, Joop J A; Messéan, Antoine; Nuti, Marco; Perry, Joe N; Sweet, Jeremy B; Tebbe, Christoph C

    2017-04-01

    In legal frameworks worldwide, genetically modified plants (GMPs) are subjected to pre-market environmental risk assessment (ERA) with the aim of identifying potential effects on the environment. In the European Union, the EFSA Guidance Document introduces the rationale that GMPs, as well as their newly produced metabolites, represent the potential stressor to be evaluated during ERA. As a consequence, during several phases of ERA for cultivation purposes, it is considered necessary to use whole plants or plant parts in experimental protocols. The importance of in planta studies as a strategy to address impacts of GMPs on non-target organisms is demonstrated, to evaluate both effects due to the intended modification in plant phenotype (e.g. expression of Cry proteins) and effects due to unintended modifications in plant phenotype resulting from the transformation process (e.g. due to somaclonal variations or pleiotropic effects). In planta tests are also necessary for GMPs in which newly expressed metabolites cannot easily be studied in vitro. This paper reviews the scientific literature supporting the choice of in planta studies as a fundamental tool in ERA of GMPs in cultivation dossiers; the evidence indicates they can realistically mimic the ecological relationships occurring in their receiving environments and provide important insights into the biology and sustainable management of GMPs.

  19. Assessment of factors influencing the Agrobacterium-mediated in planta seed transformation of brinjal (Solanum melongena L.).

    PubMed

    Subramanyam, Kondeti; Rajesh, Manoharan; Jaganath, Balusamy; Vasuki, Amirthalingam; Theboral, Jeevaraj; Elayaraja, Dhandapani; Karthik, Sivabalan; Manickavasagam, Markandan; Ganapathi, Andy

    2013-09-01

    An efficient and reproducible in planta transformation method was developed for brinjal using seed as an explant. The brinjal seeds were infected with Agrobacterium tumefaciens EHA 105 harbouring pCAMBIA 1301-bar plasmid, and the transformants were selected against BASTA®. Several parameters influencing the in planta seed transformation such as pre-culture duration, acetosyringone concentration, surfactants, duration of sonication, vacuum pressure and vacuum duration have been evaluated. The putatively transformed (T 0) brinjal plants were screened by GUS histochemical analysis. Among the different combinations and concentrations tested, when the 18-h pre-cultured brinjal seeds were sonicated for 20 min and vacuum infiltered for 3 min at 500 mm of Hg in Agrobacterium suspension containing 100 μM acetosyringone, 0.2 % Silwett L-77 favoured the Agrobacterium infection and showed maximum transformation efficiency. Among the five brinjal varieties evaluated, Arka Samhitha showed maximum transformation efficiency at 45.66 %. The transgene was successfully transmitted to progeny plants (T 1) which was evidenced by GUS histochemical analysis, polymerase chain reaction and Southern hybridisation. The in planta protocol developed in the present study would be beneficial to transfer the economically and nutritionally important genes into different varieties of brinjal, and the transgenic brinjal plants can be produced in less time (approximately 27 days).

  20. An efficient in planta transformation of Jatropha curcas (L.) and multiplication of transformed plants through in vivo grafting.

    PubMed

    Jaganath, Balusamy; Subramanyam, Kondeti; Mayavan, Subramanian; Karthik, Sivabalan; Elayaraja, Dhandapani; Udayakumar, Rajangam; Manickavasagam, Markandan; Ganapathi, Andy

    2014-05-01

    An efficient and reproducible Agrobacterium-mediated in planta transformation was developed in Jatropha curcas. The various factors affecting J. curcas in planta transformation were optimized, including decapitation, Agrobacterium strain, pin-pricking, vacuum infiltration duration and vacuum pressure. Simple vegetative in vivo cleft grafting method was adopted in the multiplication of transformants without the aid of tissue culture. Among the various parameters evaluated, decapitated plants on pin-pricking and vacuum infiltrated at 250 mmHg for 3 min with the Agrobacterium strain EHA 105 harbouring the binary vector pGA 492 was proved to be efficient in all terms with a transformation efficiency of 62.66%. Transgene integration was evinced by the GUS histochemical analysis, and the GUS positive plants were subjected to grafting. Putatively transformed J. curcas served as "Scion" and the wild type J. curcas plant severed as "Stock". There was no occurrence of graft rejection and the plants were then confirmed by GUS histochemical analysis, polymerase chain reaction (PCR) and Southern hybridization. Genetic stability of the grafted plants was evaluated by using randomly amplified polymorphic DNA (RAPD), marker which showed 100% genetic stability between mother and grafted plants. Thus, an efficient in planta transformation and grafting based multiplication of J. curcas was established.

  1. Understanding colonization and proliferation potential of endophytes and pathogen in planta via plating, polymerase chain reaction and ergosterol assay.

    PubMed

    Chow, Yiing Yng; Rahman, Sadequr; Ting, Adeline Su Yien

    2017-01-01

    This study aimed to establish the colonization behavior and proliferation potential of three endophytes and one pathogen Ganoderma boninense (Gb) introduced into oil palm ramets (host model). The endophytes selected were Diaporthe phaseolorum (WAA02), Trichoderma asperellum (T2), and Penicillium citrinum (BTF08). Ramets were first inoculated with 100 mL of fungal cells (10(6) cfu mL(-)(1)) via soil drenching. For the next 7 days, ramets were sampled and subjected to three different assays to detect and identify fungal colonization, and establish their proliferation potential in planta. Plate assay revealed the presence of endophytes in root, stem and leaf tissues within 7 days after inoculation. Polymerase Chain Reaction (PCR) detected and identified the isolates from the plant tissues. The ergosterol assay (via high performance liquid chromatography, HPLC) confirmed the presence of endophytes and Gb in planta. The increase in ergosterol levels throughout 49 days was however insignificant, suggesting that proliferation may be absent or may occur very slowly in planta. This study strongly suggests that the selected endophytes could colonize the host upon inoculation, but proliferation occurs at a slower rate, which may subsequently influence the biocontrol expression of endophytes against the pathogen.

  2. Analysis of the in planta transcriptome expressed by the corn pathogen Pantoea stewartii subsp. stewartii via RNA-Seq.

    PubMed

    Packard, Holly; Kernell Burke, Alison; Jensen, Roderick V; Stevens, Ann M

    2017-01-01

    Pantoea stewartii subsp. stewartii is a bacterial phytopathogen that causes Stewart's wilt disease in corn. It uses quorum sensing to regulate expression of some genes involved in virulence in a cell density-dependent manner as the bacterial population grows from small numbers at the initial infection site in the leaf apoplast to high cell numbers in the xylem where it forms a biofilm. There are also other genes important for pathogenesis not under quorum-sensing control such as a Type III secretion system. The purpose of this study was to compare gene expression during an in planta infection versus either a pre-inoculum in vitro liquid culture or an in vitro agar plate culture to identify genes specifically expressed in planta that may also be important for colonization and/or virulence. RNA was purified from each sample type to determine the transcriptome via RNA-Seq using Illumina sequencing of cDNA. Fold gene expression changes in the in planta data set in comparison to the two in vitro grown samples were determined and a list of the most differentially expressed genes was generated to elucidate genes important for plant association. Quantitative reverse transcription PCR (qRT-PCR) was used to validate expression patterns for a select subset of genes. Analysis of the transcriptome data via gene ontology revealed that bacterial transporters and systems important for oxidation reduction processes appear to play a critical role for P. stewartii as it colonizes and causes wilt disease in corn plants.

  3. A transient expression assay for the in planta efficacy screening of an antimicrobial peptide against grapevine bacterial pathogens.

    PubMed

    Visser, M; Stephan, D; Jaynes, J M; Burger, J T

    2012-06-01

    Natural and synthetic antimicrobial peptides (AMPs) are of increasing interest as potential resistance conferring elements in plants against pathogen infection. The efficacy of AMPs against pathogens is prescreened by in vitro assays, and promising AMP candidates are introduced as transgenes into plants. As in vitro and in planta environments differ, a prescreening procedure of the AMP efficacy in the plant environment is desired. Here, we report the efficacy of the purified synthetic peptide D4E1 against the grapevine-infecting bacterial pathogens Agrobacterium vitis and Xylophilus ampelinus in vitro and describe for the first time an in planta prescreening procedure based on transiently expressed D4E1. The antimicrobial effect of D4E1 against Ag. vitis and X. ampelinus was shown by a reduction in colony-forming units in vitro in a traditional plate-based assay and by a reduction in bacterial titres in planta as measured by quantitative real-time PCR (qPCR) in grapevine leaves transiently expressing D4E1. A statistically significant reduction in titre was shown for X. ampelinus, but for Ag. vitis, a significant reduction in titre was only observed in a subset of plants. The titres of both grapevine-infecting bacterial pathogens were reduced in an in vitro assay and for X. ampelinus in an in planta assay by D4E1 application. This widens the applicability of D4E1 as a potential resistance-enhancing element to additional pathogens and in a novel plant species. D4E1 is a promising candidate to confer enhanced resistance against the two tested grapevine bacterial pathogens, and the applied transient expression system proved to be a valuable tool for prescreening of D4E1 efficacy in an in planta environment. The described prescreening procedure can be used for other AMPs and might be adapted to other plant species and pathogens before the expensive and tedious development of stably transgenic lines is started. © 2012 The Authors. Letters in Applied Microbiology © 2012

  4. Abscisic acid metabolizing rhizobacteria decrease ABA concentrations in planta and alter plant growth.

    PubMed

    Belimov, Andrey A; Dodd, Ian C; Safronova, Vera I; Dumova, Valentina A; Shaposhnikov, Alexander I; Ladatko, Alexander G; Davies, William J

    2014-01-01

    Although endogenous phytohormones such as abscisic acid (ABA) regulate root growth, and many rhizobacteria can modulate root phytohormone status, hitherto there have been no reports of rhizobacteria mediating root ABA concentrations and growth by metabolising ABA. Using a selective ABA-supplemented medium, two bacterial strains were isolated from the rhizosphere of rice (Oryza sativa) seedlings grown in sod-podzolic soil and assigned to Rhodococcus sp. P1Y and Novosphingobium sp. P6W using partial 16S rRNA gene sequencing and phenotypic patterns by the GEN III MicroPlate test. Although strain P6W had more rapid growth in ABA-supplemented media than strain P1Y, both could utilize ABA as a sole carbon source in batch culture. When rice seeds were germinated on filter paper in association with bacteria, root ABA concentration was not affected, but shoot ABA concentration of inoculated plants decreased by 14% (strain P6W) and 22% (strain P1Y). When tomato (Solanum lycopersicum) genotypes differing in ABA biosynthesis (ABA deficient mutants flacca - flc, and notabilis - not and the wild-type cv. Ailsa Craig, WT) were grown in gnotobiotic cultures on nutrient solution agar, rhizobacterial inoculation decreased root and/or leaf ABA concentrations, depending on plant and bacteria genotypes. Strain P6W inhibited primary root elongation of all genotypes, but increased leaf biomass of WT plants. In WT plants treated with silver ions that inhibit ethylene perception, both ABA-metabolising strains significantly decreased root ABA concentration, and strain P6W decreased leaf ABA concentration. Since these changes in ABA status also occurred in plants that were not treated with silver, it suggests that ethylene was probably not involved in regulating bacteria-mediated changes in ABA concentration. Correlations between plant growth and ABA concentrations in planta suggest that ABA-metabolising rhizobacteria may stimulate growth via an ABA-dependent mechanism.

  5. Bioinspired engineering study of Plantae vascules for self-healing composite structures

    PubMed Central

    Trask, R. S.; Bond, I. P.

    2010-01-01

    This paper presents the first conceptual study into creating a Plantae-inspired vascular network within a fibre-reinforced polymer composite laminate, which provides an ongoing self-healing functionality without incurring a mass penalty. Through the application of a ‘lost-wax’ technique, orthogonal hollow vascules, inspired by the ‘ray cell’ structures found in ring porous hardwoods, were successfully introduced within a carbon fibre-reinforced epoxy polymer composite laminate. The influence on fibre architecture and mechanical behaviour of single vascules (located on the laminate centreline) when aligned parallel and transverse to the local host ply was characterized experimentally using a compression-after-impact test methodology. Ultrasonic C-scanning and high-resolution micro-CT X-ray was undertaken to identify the influence of and interaction between the internal vasculature and impact damage. The results clearly show that damage morphology is influenced by vascule orientation and that a 10 J low-velocity impact damage event is sufficient to breach the vasculature; a prerequisite for any subsequent self-healing function. The residual compressive strength after a 10 J impact was found to be dependent upon vascule orientation. In general, residual compressive strength decreased to 70 per cent of undamaged strength when vasculature was aligned parallel to the local host ply and a value of 63 per cent when aligned transverse. This bioinspired engineering study has illustrated the potential that a vasculature concept has to offer in terms of providing a self-healing function with minimum mass penalty, without initiating premature failure within a composite structure. PMID:19955122

  6. In planta comparative transcriptomics of host-adapted strains of Ralstonia solanacearum

    PubMed Central

    Ailloud, Florent; Lowe, Tiffany M.; Robène, Isabelle; Cruveiller, Stéphane; Allen, Caitilyn

    2016-01-01

    Background. Ralstonia solanacearum is an economically important plant pathogen with an unusually large host range. The Moko (banana) and NPB (not pathogenic to banana) strain groups are closely related but are adapted to distinct hosts. Previous comparative genomics studies uncovered very few differences that could account for the host range difference between these pathotypes. To better understand the basis of this host specificity, we used RNAseq to profile the transcriptomes of an R. solanacearum Moko strain and an NPB strain under in vitro and in planta conditions. Results. RNAs were sequenced from bacteria grown in rich and minimal media, and from bacteria extracted from mid-stage infected tomato, banana and melon plants. We computed differential expression between each pair of conditions to identify constitutive and host-specific gene expression differences between Moko and NPB. We found that type III secreted effectors were globally up-regulated upon plant cell contact in the NPB strain compared with the Moko strain. Genes encoding siderophore biosynthesis and nitrogen assimilation genes were highly up-regulated in the NPB strain during melon pathogenesis, while denitrification genes were up-regulated in the Moko strain during banana pathogenesis. The relatively lower expression of oxidases and the denitrification pathway during banana pathogenesis suggests that R. solanacearum experiences higher oxygen levels in banana pseudostems than in tomato or melon xylem. Conclusions. This study provides the first report of differential gene expression associated with host range variation. Despite minimal genomic divergence, the pathogenesis of Moko and NPB strains is characterized by striking differences in expression of virulence- and metabolism-related genes. PMID:26788428

  7. Cytokine changes in gastric and colonic epithelial cell in response to Planta ovata extract.

    PubMed

    Yakoob, Javed; Jafri, Wasim; Mehmood, Malik Hassan; Abbas, Zaigham; Tariq, Kanwal

    2017-03-22

    Background Psyllium (Planta ovata, Ispaghul) seed and husk are used for treatment of altered bowel habit, i. e. constipation and diarrhea. We studied the effect of Ispaghul extract on secretion of interleukin-1 beta (IL-1β) by AGS (ATCC CRL 1739) and SW480 (ATCC CCL-227) epithelial cell lines and determined whether Ispaghul extract has an effect on IL-1β secretion by Helicobacter pylori (H. pylori)-stimulated AGS cell and Escherichia coli K-12 (E. coli K-12)-stimulated SW480 cells in vitro. Methods The AGS cells and SW480 cells were pretreated with Ispaghul extract in concentrations, i. e. 3.5 and 7 μg/mL prior to infection with H. pylori and E. coli K-12. Results DNA fragmentation in AGS and SW480 cells treated with Ispaghul extract was not significant (2.3±0.8 %) compared with untreated cells (2.2±0.6 %). Ispaghul extract decreased the H. pylori-stimulated secretion of IL-1β by AGS cell (p<0.0001). This effect did not increase as the concentration of extract was increased. Ispaghul extract also decreased E. coli K-12-stimulated IL-1β secretion by SW480 cell (p<0.0001). This effect increased as the concentration of extracts was increased. Conclusions Ispaghul extract had an effect on stimulated secretion of IL-1β by the AGS and SW480 cell. It decreased pro-inflammatory reaction from both cell lines stimulated by bacteria.

  8. In planta comparative transcriptomics of host-adapted strains of Ralstonia solanacearum.

    PubMed

    Ailloud, Florent; Lowe, Tiffany M; Robène, Isabelle; Cruveiller, Stéphane; Allen, Caitilyn; Prior, Philippe

    2016-01-01

    Background. Ralstonia solanacearum is an economically important plant pathogen with an unusually large host range. The Moko (banana) and NPB (not pathogenic to banana) strain groups are closely related but are adapted to distinct hosts. Previous comparative genomics studies uncovered very few differences that could account for the host range difference between these pathotypes. To better understand the basis of this host specificity, we used RNAseq to profile the transcriptomes of an R. solanacearum Moko strain and an NPB strain under in vitro and in planta conditions. Results. RNAs were sequenced from bacteria grown in rich and minimal media, and from bacteria extracted from mid-stage infected tomato, banana and melon plants. We computed differential expression between each pair of conditions to identify constitutive and host-specific gene expression differences between Moko and NPB. We found that type III secreted effectors were globally up-regulated upon plant cell contact in the NPB strain compared with the Moko strain. Genes encoding siderophore biosynthesis and nitrogen assimilation genes were highly up-regulated in the NPB strain during melon pathogenesis, while denitrification genes were up-regulated in the Moko strain during banana pathogenesis. The relatively lower expression of oxidases and the denitrification pathway during banana pathogenesis suggests that R. solanacearum experiences higher oxygen levels in banana pseudostems than in tomato or melon xylem. Conclusions. This study provides the first report of differential gene expression associated with host range variation. Despite minimal genomic divergence, the pathogenesis of Moko and NPB strains is characterized by striking differences in expression of virulence- and metabolism-related genes.

  9. A real-time fluorogenic assay for the visualization of glycoside hydrolase activity in planta.

    PubMed

    Ibatullin, Farid M; Banasiak, Alicja; Baumann, Martin J; Greffe, Lionel; Takahashi, Junko; Mellerowicz, Ewa J; Brumer, Harry

    2009-12-01

    There currently exists a diverse array of molecular probes for the in situ localization of polysaccharides, nucleic acids, and proteins in plant cells, including reporter enzyme strategies (e.g. protein-glucuronidase fusions). In contrast, however, there is a paucity of methods for the direct analysis of endogenous glycoside hydrolases and transglycosidases responsible for cell wall remodeling. To exemplify the potential of fluorogenic resorufin glycosides to address this issue, a resorufin beta-glycoside of a xylogluco-oligosaccharide (XXXG-beta-Res) was synthesized as a specific substrate for in planta analysis of XEH activity. The resorufin aglycone is particularly distinguished for high sensitivity in muro assays due to a low pK(a) (5.8) and large extinction coefficient (epsilon 62,000 M(-1) cm(-1)), long-wavelength fluorescence (excitation 571 nm/emission 585 nm), and high quantum yield (0.74) of the corresponding anion. In vitro analyses demonstrated that XXXG-beta-Res is hydrolyzed by the archetypal plant XEH, nasturtium (Tropaeolum majus) NXG1, with classical Michaelis-Menten substrate saturation kinetics and a linear dependence on both enzyme concentration and incubation time. Further, XEH activity could be visualized in real time by observing the localized increase in fluorescence in germinating nasturtium seeds and Arabidopsis (Arabidopsis thaliana) inflorescent stems by confocal microscopy. Importantly, this new in situ XEH assay provides an essential complement to the in situ xyloglucan endotransglycosylase assay, thus allowing delineation of the disparate activities encoded by xyloglucan endotransglycosylase/hydrolase genes directly in plant tissues. The observation that XXXG-beta-Res is also hydrolyzed by diverse microbial XEHs indicates that this substrate, and resorufin glycosides in general, may find broad applicability for the analysis of wall restructuring by polysaccharide hydrolases during morphogenesis and plant-microbe interactions.

  10. Bioinspired engineering study of Plantae vascules for self-healing composite structures.

    PubMed

    Trask, R S; Bond, I P

    2010-06-06

    This paper presents the first conceptual study into creating a Plantae-inspired vascular network within a fibre-reinforced polymer composite laminate, which provides an ongoing self-healing functionality without incurring a mass penalty. Through the application of a 'lost-wax' technique, orthogonal hollow vascules, inspired by the 'ray cell' structures found in ring porous hardwoods, were successfully introduced within a carbon fibre-reinforced epoxy polymer composite laminate. The influence on fibre architecture and mechanical behaviour of single vascules (located on the laminate centreline) when aligned parallel and transverse to the local host ply was characterized experimentally using a compression-after-impact test methodology. Ultrasonic C-scanning and high-resolution micro-CT X-ray was undertaken to identify the influence of and interaction between the internal vasculature and impact damage. The results clearly show that damage morphology is influenced by vascule orientation and that a 10 J low-velocity impact damage event is sufficient to breach the vasculature; a prerequisite for any subsequent self-healing function. The residual compressive strength after a 10 J impact was found to be dependent upon vascule orientation. In general, residual compressive strength decreased to 70 per cent of undamaged strength when vasculature was aligned parallel to the local host ply and a value of 63 per cent when aligned transverse. This bioinspired engineering study has illustrated the potential that a vasculature concept has to offer in terms of providing a self-healing function with minimum mass penalty, without initiating premature failure within a composite structure.

  11. RNA-guided transcriptional regulation in planta via synthetic dCas9-based transcription factors.

    PubMed

    Piatek, Agnieszka; Ali, Zahir; Baazim, Hatoon; Li, Lixin; Abulfaraj, Aala; Al-Shareef, Sahar; Aouida, Mustapha; Mahfouz, Magdy M

    2015-05-01

    Targeted genomic regulation is a powerful approach to accelerate trait discovery and development in agricultural biotechnology. Bacteria and archaea use clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated (Cas) regulatory systems for adaptive molecular immunity against foreign nucleic acids introduced by invading phages and conjugative plasmids. The type II CRISPR/Cas system has been adapted for genome editing in many cell types and organisms. A recent study used the catalytically inactive Cas9 (dCas9) protein combined with guide-RNAs (gRNAs) as a DNA-targeting platform to modulate gene expression in bacterial, yeast, and human cells. Here, we modified this DNA-targeting platform for targeted transcriptional regulation in planta by developing chimeric dCas9-based transcriptional activators and repressors. To generate transcriptional activators, we fused the dCas9 C-terminus with the activation domains of EDLL and TAL effectors. To generate a transcriptional repressor, we fused the dCas9 C-terminus with the SRDX repression domain. Our data demonstrate that dCas9 fusion with the EDLL activation domain (dCas9:EDLL) and the TAL activation domain (dCas9:TAD), guided by gRNAs complementary to selected promoter elements, induce strong transcriptional activation on Bs3::uidA targets in plant cells. Further, the dCas9:SRDX-mediated transcriptional repression of an endogenous gene. Thus, our results suggest that the synthetic transcriptional repressor (dCas9:SRDX) and activators (dCas9:EDLL and dCas9:TAD) can be used as endogenous transcription factors to repress or activate transcription of an endogenous genomic target. Our data indicate that the CRISPR/dCas9 DNA-targeting platform can be used in plants as a functional genomics tool and for biotechnological applications. © 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  12. Floral benzenoid carboxyl methyltransferases: From in vitro to in planta function

    PubMed Central

    Effmert, Uta; Saschenbrecker, Sandra; Ross, Jeannine; Negre, Florence; Fraser, Chris M.; Noel, Joseph P.; Dudareva, Natalia; Piechulla, Birgit

    2010-01-01

    Benzenoid carboxyl methyltransferases synthesize methyl esters (e.g., methyl benzoate and methyl salicylate), which are constituents of aromas and scents of many plant species and play important roles in plant communication with the surrounding environment. Within the past five years, eleven such carboxyl methyltransferases were isolated and most of them were comprehensively investigated at the biochemical, molecular and structural level. Two types of enzymes can be distinguished according to their substrate preferences: the SAMT-type enzymes isolated from Clarkia breweri, Stephanotis floribunda, Antirrhinum majus, Hoya carnosa, and Petunia hybrida, which have a higher catalytic efficiency and preference for salicylic acid, while BAMT-type enzymes from A. majus, Arabidopsis thaliana, Arabidopsis lyrata, and Nicotiana suaveolens prefer benzoic acid. The elucidation of C. breweri SAMT’s three-dimensional structure allowed a detailed modelling of the active sites of the carboxyl methyltransferases and revealed that the SAM binding pocket is highly conserved among these enzymes while the methyl acceptor binding site exhibits some variability, allowing a classification into SAMT-type and BAMT-type enzymes. The analysis of expression patterns coupled with biochemical characterization showed that these carboxyl methyltransferases are involved either in floral scent biosynthesis or in plant defense responses. While the latter can be induced by biotic or abiotic stress, the genes responsible for floral scent synthesis exhibit developmental and rhythmic expression pattern. The nature of the product and efficiency of its formation in planta depend on the availability of substrates, the catalytic efficiency of the enzyme toward benzoic acid and/or salicylic acid, and the transcriptional, translational, and post-translational regulation at the enzyme level. The biochemical properties of benzenoid carboxyl methyltransferases suggest that the genes involved in plant defenses

  13. A rapid assay to quantify the cleavage efficiency of custom-designed nucleases in planta.

    PubMed

    Johnson, Ross A; Gurevich, Vyacheslav; Levy, Avraham A

    2013-06-01

    Custom-designed nucleases are a promising technology for genome editing through the catalysis of double-strand DNA breaks within target loci and subsequent repair by the host cell, which can result in targeted mutagenesis or gene replacement. Implementing this new technology requires a rapid means to determine the cleavage efficiency of these custom-designed proteins in planta. Here we present such an assay that is based on cleavage-dependent luciferase gene correction as part of a transient dual-luciferase(®) reporter (Promega) expression system. This assay consists of co-infiltrating Nicotiana benthamiana leaves with two Agrobacterium tumefaciens strains: one contains the target sequence embedded within a luciferase reporter gene and the second strain contains the custom-designed nuclease gene(s). We compared repair following site-specific nuclease digestion through non-homologous DNA end-joining, as opposed to single strand DNA annealing, as a means to restore an out-of-frame luciferase gene cleavage-reporter construct. We show, using luminometer measurements and bioluminescence imaging, that the assay for non-homologous end-joining is sensitive, quantitative, reproducible and rapid in estimating custom-designed nucleases' cleavage efficiency. We detected cleavage by two out of three transcription activator-like effector nucleases that we custom-designed for targets in the Arabidopsis CRUCIFERIN3 gene, and we compared with the well-established 'QQR' zinc-finger nuclease. The assay we report requires only standard equipment and basic plant molecular biology techniques, and it can be carried out within a few days. Different types of custom-designed nucleases can be preliminarily tested in our assay system before their downstream application in plant genome editing.

  14. Label-free chemically specific imaging in planta with stimulated Raman scattering microscopy.

    PubMed

    Mansfield, Jessica C; Littlejohn, George R; Seymour, Mark P; Lind, Rob J; Perfect, Sarah; Moger, Julian

    2013-05-21

    The growing world population puts ever-increasing demands on the agricultural and agrochemical industries to increase agricultural yields. This can only be achieved by investing in fundamental plant and agrochemical research and in the development of improved analytical tools to support research in these areas. There is currently a lack of analytical tools that provide noninvasive structural and chemical analysis of plant tissues at the cellular scale. Imaging techniques such as coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS) microscopy provide label-free chemically specific image contrast based on vibrational spectroscopy. Over the past decade, these techniques have been shown to offer clear advantages for a vast range of biomedical research applications. The intrinsic vibrational contrast provides label-free quantitative functional analysis, it does not suffer from photobleaching, and it allows near real-time imaging in 3D with submicrometer spatial resolution. However, due to the susceptibility of current detection schemes to optical absorption and fluorescence from pigments (such as chlorophyll), the plant science and agrochemical research communities have not been able to benefit from these techniques and their application in plant research has remained virtually unexplored. In this paper, we explore the effect of chlorophyll fluorescence and absorption in CARS and SRS microscopy. We show that with the latter it is possible to use phase-sensitive detection to separate the vibrational signal from the (electronic) absorption processes. Finally, we demonstrate the potential of SRS for a range of in planta applications by presenting in situ chemical analysis of plant cell wall components, epicuticular waxes, and the deposition of agrochemical formulations onto the leaf surface.

  15. The In Planta Transcriptome of Ralstonia solanacearum: Conserved Physiological and Virulence Strategies during Bacterial Wilt of Tomato

    PubMed Central

    Jacobs, Jonathan M.; Babujee, Lavanya; Meng, Fanhong; Milling, Annett; Allen, Caitilyn

    2012-01-01

    ABSTRACT Plant xylem fluid is considered a nutrient-poor environment, but the bacterial wilt pathogen Ralstonia solanacearum is well adapted to it, growing to 108 to 109 CFU/g tomato stem. To better understand how R. solanacearum succeeds in this habitat, we analyzed the transcriptomes of two phylogenetically distinct R. solanacearum strains that both wilt tomato, strains UW551 (phylotype II) and GMI1000 (phylotype I). We profiled bacterial gene expression at ~6 × 108 CFU/ml in culture or in plant xylem during early tomato bacterial wilt pathogenesis. Despite phylogenetic differences, these two strains expressed their 3,477 common orthologous genes in generally similar patterns, with about 12% of their transcriptomes significantly altered in planta versus in rich medium. Several primary metabolic pathways were highly expressed during pathogenesis. These pathways included sucrose uptake and catabolism, and components of these pathways were encoded by genes in the scrABY cluster. A UW551 scrA mutant was significantly reduced in virulence on resistant and susceptible tomato as well as on potato and the epidemiologically important weed host Solanum dulcamara. Functional scrA contributed to pathogen competitive fitness during colonization of tomato xylem, which contained ~300 µM sucrose. scrA expression was induced by sucrose, but to a much greater degree by growth in planta. Unexpectedly, 45% of the genes directly regulated by HrpB, the transcriptional activator of the type 3 secretion system (T3SS), were upregulated in planta at high cell densities. This result modifies a regulatory model based on bacterial behavior in culture, where this key virulence factor is repressed at high cell densities. The active transcription of these genes in wilting plants suggests that T3SS has a biological role throughout the disease cycle. PMID:22807564

  16. Significant reduction of BiFC non-specific assembly facilitates in planta assessment of heterotrimeric G-protein interactors.

    PubMed

    Gookin, Timothy E; Assmann, Sarah M

    2014-11-01

    Protein networks and signaling cascades are key mechanisms for intra- and intercellular signal transduction. Identifying the interacting partners of a protein can provide vital clues regarding its physiological role. The bimolecular fluorescence complementation (BiFC) assay has become a routine tool for in vivo analysis of protein-protein interactions and their subcellular location. Although the BiFC system has improved since its inception, the available options for in planta analysis are still subject to very low signal-to-noise ratios, and a systematic comparison of BiFC confounding background signals has been lacking. Background signals can obscure weak interactions, provide false positives, and decrease confidence in true positives. To overcome these problems, we performed an extensive in planta analysis of published BiFC fragments used in metazoa and plants, and then developed an optimized single vector BiFC system which utilizes monomeric Venus (mVenus) split at residue 210, and contains an integrated mTurquoise2 marker to precisely identify transformed cells in order to distinguish true negatives. Here we provide our streamlined double ORF expression (pDOE) BiFC system, and show that our advance in BiFC methodology functions even with an internally fused mVenus210 fragment. We illustrate the efficacy of the system by providing direct visualization of Arabidopsis MLO1 interacting with a calmodulin-like (CML) protein, and by showing that heterotrimeric G-protein subunits Gα (GPA1) and Gβ (AGB1) interact in plant cells. We further demonstrate that GPA1 and AGB1 each physically interact with PLDα1 in planta, and that mutation of the so-called PLDα1 'DRY' motif abolishes both of these interactions. © 2014 The Authors The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

  17. Analysis of the in planta transcriptome expressed by the corn pathogen Pantoea stewartii subsp. stewartii via RNA-Seq

    PubMed Central

    Packard, Holly; Kernell Burke, Alison; Jensen, Roderick V.

    2017-01-01

    Pantoea stewartii subsp. stewartii is a bacterial phytopathogen that causes Stewart’s wilt disease in corn. It uses quorum sensing to regulate expression of some genes involved in virulence in a cell density-dependent manner as the bacterial population grows from small numbers at the initial infection site in the leaf apoplast to high cell numbers in the xylem where it forms a biofilm. There are also other genes important for pathogenesis not under quorum-sensing control such as a Type III secretion system. The purpose of this study was to compare gene expression during an in planta infection versus either a pre-inoculum in vitro liquid culture or an in vitro agar plate culture to identify genes specifically expressed in planta that may also be important for colonization and/or virulence. RNA was purified from each sample type to determine the transcriptome via RNA-Seq using Illumina sequencing of cDNA. Fold gene expression changes in the in planta data set in comparison to the two in vitro grown samples were determined and a list of the most differentially expressed genes was generated to elucidate genes important for plant association. Quantitative reverse transcription PCR (qRT-PCR) was used to validate expression patterns for a select subset of genes. Analysis of the transcriptome data via gene ontology revealed that bacterial transporters and systems important for oxidation reduction processes appear to play a critical role for P. stewartii as it colonizes and causes wilt disease in corn plants. PMID:28462040

  18. Towards an Understanding of Enhanced Biomass Digestibility by In Planta Expression of a Family 5 Glycoside Hydrolase

    DOE PAGES

    Donohoe, Bryon S.; Wei, Hui; Mittal, Ashutosh; ...

    2017-06-29

    In planta expression of a thermophilic endoglucanase (AcCel5A) reduces recalcitrance by creating voids and other irregularities in cell walls of Arabidopsis thaliana that increase enzyme accessibility without negative impacts on plant growth or cell wall composition. Furthermore, our results suggest that cellulose β-1–4 linkages can be cut sparingly in the assembling wall and that these minimal changes, made at the proper time, have an impact on plant cell wall recalcitrance without negative effects on overall plant development.

  19. Nodularin, a cyanobacterial toxin, is synthesized in planta by symbiotic Nostoc sp.

    PubMed Central

    Gehringer, Michelle M; Adler, Lewis; Roberts, Alexandra A; Moffitt, Michelle C; Mihali, Troco K; Mills, Toby J T; Fieker, Claus; Neilan, Brett A

    2012-01-01

    The nitrogen-fixing bacterium, Nostoc, is a commonly occurring cyanobacterium often found in symbiotic associations. We investigated the potential of cycad cyanobacterial endosymbionts to synthesize microcystin/nodularin. Endosymbiont DNA was screened for the aminotransferase domain of the toxin biosynthesis gene clusters. Five endosymbionts carrying the gene were screened for bioactivity. Extracts of two isolates inhibited protein phosphatase 2A and were further analyzed using electrospray ionization mass spectrometry (ESI-MS)/MS. Nostoc sp. ‘Macrozamia riedlei 65.1' and Nostoc sp. ‘Macrozamia serpentina 73.1' both contained nodularin. High performance liquid chromatography (HPLC) HESI-MS/MS analysis confirmed the presence of nodularin at 9.55±2.4 ng μg−1 chlorophyll a in Nostoc sp. ‘Macrozamia riedlei 65.1' and 12.5±8.4 ng μg−1 Chl a in Nostoc sp. ‘Macrozamia serpentina 73.1' extracts. Further scans indicated the presence of the rare isoform [L-Har2] nodularin, which contains ℒ-homoarginine instead of ℒ-arginine. Nodularin was also present at 1.34±0.74 ng ml−1 (approximately 3 pmol per g plant ww) in the methanol root extracts of M. riedlei MZ65, while the presence of [L-Har2] nodularin in the roots of M. serpentina MZ73 was suggested by HPLC HESI-MS/MS analysis. The ndaA-B and ndaF genomic regions were sequenced to confirm the presence of the hybrid polyketide/non-ribosomal gene cluster. A seven amino-acid insertion into the NdaA-C1 domain of N. spumigena NSOR10 protein was observed in all endosymbiont-derived sequences, suggesting the transfer of the nda cluster from N. spumigena to terrestrial Nostoc species. This study demonstrates the synthesis of nodularin and [L-Har2] nodularin in a non-Nodularia species and the production of cyanobacterial hepatotoxin by a symbiont in planta. PMID:22456448

  20. Nodularin, a cyanobacterial toxin, is synthesized in planta by symbiotic Nostoc sp.

    PubMed

    Gehringer, Michelle M; Adler, Lewis; Roberts, Alexandra A; Moffitt, Michelle C; Mihali, Troco K; Mills, Toby J T; Fieker, Claus; Neilan, Brett A

    2012-10-01

    The nitrogen-fixing bacterium, Nostoc, is a commonly occurring cyanobacterium often found in symbiotic associations. We investigated the potential of cycad cyanobacterial endosymbionts to synthesize microcystin/nodularin. Endosymbiont DNA was screened for the aminotransferase domain of the toxin biosynthesis gene clusters. Five endosymbionts carrying the gene were screened for bioactivity. Extracts of two isolates inhibited protein phosphatase 2A and were further analyzed using electrospray ionization mass spectrometry (ESI-MS)/MS. Nostoc sp. 'Macrozamia riedlei 65.1' and Nostoc sp. 'Macrozamia serpentina 73.1' both contained nodularin. High performance liquid chromatography (HPLC) HESI-MS/MS analysis confirmed the presence of nodularin at 9.55±2.4 ng μg-1 chlorophyll a in Nostoc sp. 'Macrozamia riedlei 65.1' and 12.5±8.4 ng μg-1 Chl a in Nostoc sp. 'Macrozamia serpentina 73.1' extracts. Further scans indicated the presence of the rare isoform [L-Har(2)] nodularin, which contains L-homoarginine instead of L-arginine. Nodularin was also present at 1.34±0.74 ng ml(-1) (approximately 3 pmol per g plant ww) in the methanol root extracts of M. riedlei MZ65, while the presence of [L-Har(2)] nodularin in the roots of M. serpentina MZ73 was suggested by HPLC HESI-MS/MS analysis. The ndaA-B and ndaF genomic regions were sequenced to confirm the presence of the hybrid polyketide/non-ribosomal gene cluster. A seven amino-acid insertion into the NdaA-C1 domain of N. spumigena NSOR10 protein was observed in all endosymbiont-derived sequences, suggesting the transfer of the nda cluster from N. spumigena to terrestrial Nostoc species. This study demonstrates the synthesis of nodularin and [L-Har(2)] nodularin in a non-Nodularia species and the production of cyanobacterial hepatotoxin by a symbiont in planta.

  1. In planta gene expression analysis of Xanthomonas oryzae pathovar oryzae, African strain MAI1

    PubMed Central

    2010-01-01

    Background Bacterial leaf blight causes significant yield losses in rice crops throughout Asia and Africa. Although both the Asian and African strains of the pathogen, Xanthomonas oryzae pv. oryzae (Xoo), induce similar symptoms, they are nevertheless genetically different, with the African strains being more closely related to the Asian X. oryzae pv. oryzicola (Xoc). Results Changes in gene expression of the African Xoo strain MAI1 in the susceptible rice cultivar Nipponbare were profiled, using an SSH Xoo DNA microarray. Microarray hybridization was performed comparing bacteria recovered from plant tissues at 1, 3, and 6 days after inoculation (dai) with bacteria grown in vitro. A total of 710 bacterial genes were found to be differentially expressed, with 407 up-regulated and 303 down-regulated. Expression profiling indicated that less than 20% of the 710 bacterial transcripts were induced in the first 24 h after inoculation, whereas 63% were differentially expressed at 6 dai. The 710 differentially expressed genes were one-end sequenced. 535 sequences were obtained from which 147 non-redundant sequences were identified. Differentially expressed genes were related to metabolism, secretion and transport, pathogen adherence to plant tissues, plant cell-wall degradation, IS elements, and virulence. In addition, various other genes encoding proteins with unknown function or showing no similarity to other proteins were also induced. The Xoo MAI1 non-redundant set of sequences was compared against several X. oryzae genomes, revealing a specific group of genes that was present only in MAI1. Numerous IS elements were also found to be differentially expressed. Quantitative real-time PCR confirmed 86% of the identified profile on a set of 14 genes selected according to the microarray analysis. Conclusions This is the first report to compare the expression of Xoo genes in planta across different time points during infection. This work shows that as-yet-unidentified and

  2. Differential subcellular targeting of recombinant human α₁-proteinase inhibitor influences yield, biological activity and in planta stability of the protein in transgenic tomato plants.

    PubMed

    Jha, Shweta; Agarwal, Saurabh; Sanyal, Indraneel; Jain, G K; Amla, D V

    2012-11-01

    The response of protein accumulation site on yield, biological activity and in planta stability of therapeutic recombinant human proteinase inhibitor (α₁-PI) was analyzed via targeting to different subcellular locations, like endoplasmic reticulum (ER), apoplast, vacuole and cytosol in leaves of transgenic tomato plants. In situ localization of the recombinant α₁-PI protein in transgenic plant cells was monitored by immunohistochemical staining. Maximum accumulation of recombinant α₁-PI in T₀ and T₁ transgenic tomato plants was achieved from 1.5 to 3.2% of total soluble protein (TSP) by retention in ER lumen, followed by vacuole and apoplast, whereas cytosolic targeting resulted into degradation of the protein. The plant-derived recombinant α₁-PI showed biological activity for elastase inhibition, as monitored by residual porcine pancreatic elastase (PPE) activity assay and band-shift assay. Recombinant α₁-PI was purified from transgenic tomato plants with high yield, homogeneity and biological activity. Purified protein appeared as a single band of ∼48-50 kDa on SDS-PAGE with pI value ranging between 5.1 and 5.3. Results of mass spectrometry and optical spectroscopy of purified recombinant α₁-PI revealed the structural integrity of the recombinant protein comparable to native serum α₁-PI. Enzymatic deglycosylation and lectin-binding assays with the purified recombinant α₁-PI showed compartment-specific N-glycosylation of the protein targeted to ER, apoplast and vacuole. Conformational studies based on urea-induced denaturation and circular dichroism (CD) spectroscopy revealed relatively lower stability of the recombinant α₁-PI protein, compared to its serum counterpart. Pharmacokinetic evaluation of plant derived recombinant and human plasma-purified α₁-PI in rat, by intravenous route, revealed significantly faster plasma clearance and lower area under curve (AUC) of recombinant protein. Our data suggested significance of

  3. Burkholderia phymatum is a highly effective nitrogen-fixing symbiont of Mimosa spp. and fixes nitrogen ex planta.

    PubMed

    Elliott, Geoffrey N; Chen, Wen-Ming; Chou, Jui-Hsing; Wang, Hui-Chun; Sheu, Shih-Yi; Perin, Liamara; Reis, Veronica M; Moulin, Lionel; Simon, Marcelo F; Bontemps, Cyril; Sutherland, Joan M; Bessi, Rosana; de Faria, Sergio M; Trinick, Michael J; Prescott, Alan R; Sprent, Janet I; James, Euan K

    2007-01-01

    * The ability of Burkholderia phymatum STM815 to effectively nodulate Mimosa spp., and to fix nitrogen ex planta, was compared with that of the known Mimosa symbiont Cupriavidus taiwanensis LMG19424. * Both strains were equally effective symbionts of M. pudica, but nodules formed by STM815 had greater nitrogenase activity. STM815 was shown to have a broader host range across the genus Mimosa than LMG19424, nodulating 30 out of 31 species, 21 of these effectively. LMG19424 effectively nodulated only nine species. GFP-marked variants were used to visualise symbiont presence within nodules. * STM815 gave significant acetylene reduction assay (ARA) activity in semisolid JMV medium ex planta, but no ARA activity was detected with LMG19424. 16S rDNA sequences of two isolates originally from Mimosa nodules in Papua New Guinea (NGR114 and NGR195A) identified them as Burkholderia phymatum also, with nodA, nodC and nifH genes of NGR195A identical to those of STM815. * B. phymatum is therefore an effective Mimosa symbiont with a broad host range, and is the first reported beta-rhizobial strain to fix nitrogen in free-living culture.

  4. In planta expression of a mature Der p 1 allergen isolated from an Italian strain of Dermatophagoides pteronyssinus.

    PubMed

    Marconi, Gianpiero; Albertini, Emidio; Mari, Adriano; Palazzo, Paola; Porceddu, Andrea; Raggi, Lorenzo; Bolis, Luigi; Lancioni, Hovirag; Palomba, Antonella; Lucentini, Livia; Lanfaloni, Luisa; Marcucci, Francesco; Falcinelli, Mario; Panara, Fausto

    2012-06-01

    European (Dermatophagoides pteronyssinus) and American (Dermatophagoides farinae) house dust mite species are considered the most common causes of asthma and allergic symptoms worldwide. Der p 1 protein, one of the main allergens of D. pteronyssinus, is found in high concentration in mites faecal pellets, which can became easily airborne and, when inhaled, can cause perennial rhinitis and bronchial asthma. Here we report the isolation of the Der p 1 gene from an Italian strain of D. pteronyssinus and the PVX-mediated expression of its mature form (I-rDer p 1) in Nicotiana benthamiana plants. Human sera from characterized allergic patients were used for IgE binding inhibition assays to test the immunological reactivity of I-rDer p 1 produced in N. benthamiana plants. The binding properties of in planta produced I-rDer p 1 versus the IgE of patients sera were comparable to those obtained on Der p 1 preparation immobilized on a microarray. In this paper we provide a proof of concept for the production of an immunologically active form of Der p 1 using a plant viral vector. These results pave the way for the development of diagnostic allergy tests based on in planta produced allergens.

  5. Characterization of in planta-induced rust genes isolated from a haustorium-specific cDNA library.

    PubMed

    Hahn, M; Mendgen, K

    1997-05-01

    Rust fungi are plant parasites that depend on living host tissue for growth. For invasion of leaves, dikaryotic urediospores differentiate germ tubes and infection structures that penetrate through stomata. Biotrophic growth occurs by intercellular mycelia that form haustoria within host cells. A cDNA library was constructed from haustoria isolated from broad bean leaves infected by Uromyces fabae. Differential screening revealed that a high proportion (19%) of the haustorial cDNAs are specifically expressed in planta but are not expressed, or are much weaker, in germlings or infection structures produced in vitro. A total of 31 different in planta-induced genes (PIGs) were identified. Some of the PIGs are highly expressed in haustoria. The PIGs are single or low copy number genes in the rust genome. A variety of developmentally regulated expression patterns of PIG mRNAs were observed. Sequence analysis of PIG cDNAs revealed similarities to genes encoding proteins involved in amino acid transport, thiamine biosynthesis, short-chain dehydrogenases, metallothioneins, cytochrome P-450 monooxygenases, and peptidyl-prolyl isomerases.

  6. In-planta sporulation phenotype: a major life history trait to understand the evolution of Alnus-infective Frankia strains.

    PubMed

    Pozzi, Adrien C; Bautista-Guerrero, Hector H; Nouioui, Imen; Cotin-Galvan, Laëtitia; Pepin, Régis; Fournier, Pascale; Menu, Frédéric; Fernandez, Maria P; Herrera-Belaroussi, Aude

    2015-09-01

    Two major types of Frankia strains are usually recognized, based on the ability to sporulate in-planta: spore-positive (Sp+) and spore-negative (Sp-). We carried out a study of Sp+ and Sp- Frankia strains based on nodules collected on Alnus glutinosa, Alnus incana and Alnus viridis. The nodules were phenotyped using improved histology methods, and endophytic Frankia strain genotype was determined using a multilocus sequence analysis approach. An additional sampling was done to assess the relation between Sp+ phenotype frequency and genetic diversity of Frankia strains at the alder stand scale. Our results revealed that (i) Sp+ and Sp- Alnus-infective Frankia strains are genetically different even when sampled from the same alder stand and the same host-plant species; (ii) there are at least two distinct phylogenetic lineages of Sp+ Frankia that cluster according to the host-plant species and without regard of geographic distance and (iii) genetic diversity of Sp+ strains is very low at the alder stand scale compared with Sp- strains. Difference in evolutionary history and genetic diversity between Sp+ and Sp- Frankia allows us to discuss the possible ecological role of in-planta sporulation. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

  7. Identification and characterization of in planta-expressed secreted effector proteins from Magnaporthe oryzae that induce cell death in rice.

    PubMed

    Chen, Songbiao; Songkumarn, Pattavipha; Venu, R C; Gowda, Malali; Bellizzi, Maria; Hu, Jinnan; Liu, Wende; Ebbole, Daniel; Meyers, Blake; Mitchell, Thomas; Wang, Guo-Liang

    2013-02-01

    Interactions between rice and Magnaporthe oryzae involve the recognition of cellular components and the exchange of complex molecular signals from both partners. How these interactions occur in rice cells is still elusive. We employed robust-long serial analysis of gene expression, massively parallel signature sequencing, and sequencing by synthesis to examine transcriptome profiles of infected rice leaves. A total of 6,413 in planta-expressed fungal genes, including 851 genes encoding predicted effector proteins, were identified. We used a protoplast transient expression system to assess 42 of the predicted effector proteins for the ability to induce plant cell death. Ectopic expression assays identified five novel effectors that induced host cell death only when they contained the signal peptide for secretion to the extracellular space. Four of them induced cell death in Nicotiana benthamiana. Although the five effectors are highly diverse in their sequences, the physiological basis of cell death induced by each was similar. This study demonstrates that our integrative genomic approach is effective for the identification of in planta-expressed cell death-inducing effectors from M. oryzae that may play an important role facilitating colonization and fungal growth during infection.

  8. The da Vinci robot.

    PubMed

    Moran, Michael E

    2006-12-01

    One might assume from the title of this paper that the nuances of a complex mechanical robot will be discussed, and this would be correct. On the other hand, the date of the design and possible construction of this robot was 1495, a little more than five centuries ago. The key point in the title is the lack of a trademarked name, as Leonardo was the designer of this sophisticated system. His notes from the Codex Altanticus represent the foundation of this report. English translations of da Vinci's notebooks are currently available. Beginning in the 1950s, investigators at the University of California began to ponder the significance of some of da Vinci's markings on what appeared to be technical drawings. Such markings also occur in his Codex Atlanticus (the largest single collection of da Vinci's sheets, consisting of 1119 separate pages and 481 folios) along with a large number of other mechanical devices. Continuing research at the Instituto e Museo di Storia della Scienza in Florence has yielded a great deal of information about Leonardo's intentions with regard to his mechanical knight. It is now known that da Vinci's robot would have had the outer appearance of a Germanic knight. It had a complex core of mechanical devices that probably was human powered. The robot had two independent operating systems. The first had three degree-of-freedom legs, ankles, knees, and hips. The second had four degrees of freedom in the arms with articulated shoulders, elbows, wrists, and hands. A mechanical analog-programmable controller within the chest provided the power and control for the arms. The legs were powered by an external crank arrangement driving the cable, which connected to key locations near each lower extremity's joints. Da Vinci also is known to have devised a programmable front-wheel-drive automobile with rack-and-pinion suspension mechanisms at age 26. He would recall this device again, when, at age 40, he is thought to have built a programmable automated

  9. A single base pair in the right terminal domain of Tomato planta macho viroid is a virulence determinant factor on tomato

    USDA-ARS?s Scientific Manuscript database

    Tomato planta macho viroid (TPMVd), including isolates previously designated as Mexican papita viroid (MPVd), causes serious disease on tomatoes in North America. Two predominant variants, sharing 93.8% sequence identity, incited distinct severe (MPVd-S) or mild (MPVd-M) symptoms on tomato. To ide...

  10. Bioimaging techniques for subcellular localization of plant hemoglobins and measurement of hemoglobin-dependent nitric oxide scavenging in planta.

    PubMed

    Hebelstrup, Kim H; Østergaard-Jensen, Erik; Hill, Robert D

    2008-01-01

    Plant hemoglobins are ubiquitous in all plant families. They are expressed at low levels in specific tissues. Several studies have established that plant hemoglobins are scavengers of nitric oxide (NO) and that varying the endogenous level of hemoglobin in plant cells negatively modulates bioactivity of NO generated under hypoxic conditions or during cellular signaling. Earlier methods for determination of hemoglobin-dependent scavenging in planta were based on measuring activity in whole plants or organs. Plant hemoglobins do not contain specific organelle localization signals; however, earlier reports on plant hemoglobin have demonstrated either cytosolic or nuclear localization, depending on the method or cell type investigated. We have developed two bioimaging techniques: one for visualization of hemoglobin-catalyzed scavenging of NO in specific cells and another for visualization of subcellular localization of green fluorescent protein-tagged plant hemoglobins in transformed Arabidopsis thaliana plants.

  11. Plastid-localized amino acid biosynthetic pathways of Plantae are predominantly composed of non-cyanobacterial enzymes

    PubMed Central

    Reyes-Prieto, Adrian; Moustafa, Ahmed

    2012-01-01

    Studies of photosynthetic eukaryotes have revealed that the evolution of plastids from cyanobacteria involved the recruitment of non-cyanobacterial proteins. Our phylogenetic survey of >100 Arabidopsis nuclear-encoded plastid enzymes involved in amino acid biosynthesis identified only 21 unambiguous cyanobacterial-derived proteins. Some of the several non-cyanobacterial plastid enzymes have a shared phylogenetic origin in the three Plantae lineages. We hypothesize that during the evolution of plastids some enzymes encoded in the host nuclear genome were mistargeted into the plastid. Then, the activity of those foreign enzymes was sustained by both the plastid metabolites and interactions with the native cyanobacterial enzymes. Some of the novel enzymatic activities were favored by selective compartmentation of additional complementary enzymes. The mosaic phylogenetic composition of the plastid amino acid biosynthetic pathways and the reduced number of plastid-encoded proteins of non-cyanobacterial origin suggest that enzyme recruitment underlies the recompartmentation of metabolic routes during the evolution of plastids. PMID:23233874

  12. Behavior of RNAi suppressor protein 2b of Cucumber mosaic virus in planta in presence and absence of virus.

    PubMed

    Praveen, Shelly; Mangrauthia, Satendra K; Singh, Priyanka; Mishra, Anil K

    2008-08-01

    The 2b protein encoded by Cucumber mosaic virus (CMV) has been shown as a virus counter defense factor that interferes with the RNAi pathway. The 2b gene from CMV-banana, New Delhi isolate (CMV-NDLS) was amplified from CMV infected cucumber plants to generate the sense and antisense binary vector constructs for 2b expression and repression in planta. Constitutive expression of 2b gene in healthy Nicotiana tabacum caused phenotypic aberrations during somatic embryogenesis, which were not observed when expressed in CMV infected N. tabacum. Further, the established virus population in CMV infected N. tabacum was not affected by constitutive expression and repression of 2b gene. Thus, indicating its role in initiation of gene silencing, at the early stage of viral infection. This is the first demonstration of differential behavior of 2b suppressor protein in host development in the absence and presence of virus.

  13. Homoserine and asparagine are host signals that trigger in planta expression of a pathogenesis gene in Nectria haematococca.

    PubMed

    Yang, Zhennai; Rogers, Linda M; Song, Yuanda; Guo, Wenjin; Kolattukudy, P E

    2005-03-15

    Some pathogenesis-related genes are expressed in fungi only when the pathogen is in the host, but the host signals that trigger these gene expressions have not been identified. Virulent Nectria haematococca infects pea plants and requires either pelA, which is induced by pectin, or pelD, which is induced only in planta. However, the host signal(s) that trigger pelD expression was unknown. Here we report the isolation of the host signals and identify homoserine and asparagine, two free amino acids found in uniquely high levels in pea seedlings, as the pelD-inducing signals. N. haematococca has evolved a mechanism to sense the host tissue environment by using the high levels of two free amino acids in this plant, thereby triggering the expression of pelD to assist the pathogenic process.

  14. Unfolding of in planta activity of anti-rep ribozyme in presence of a RNA silencing suppressor.

    PubMed

    Mishra, Sumona Karjee; Chilakamarthi, Ushasri; Deb, J K; Mukherjee, Sunil Kumar

    2014-05-21

    Antisense RNA ribozymes have intrinsic endonucleolytic activity to effect cleavage of the target RNA. However, this activity in vivo is often controlled by the dominance of antisense or other double-stranded RNA mechanism. In this work, we demonstrate the in planta activity of a hammerhead ribozyme designed to target rep-mRNA of a phytopathogen Mungbean Yellow Mosaic India virus (MYMIV) as an antiviral agent. We also found RNA-silencing is induced on introduction of catalytically active as well as inactive ribozymes. Using RNA-silencing suppressors (RSS), we demonstrate that the endonucleolytic activity of ribozymes is a true phenomenon, even while a mutated version may demonstrate a similar down-regulation of the target RNA. This helps to ease the confusion over the action mechanism of ribozymes in vivo. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  15. Homoserine and asparagine are host signals that trigger in planta expression of a pathogenesis gene in Nectria haematococca

    PubMed Central

    Yang, Zhennai; Rogers, Linda M.; Song, Yuanda; Guo, Wenjin; Kolattukudy, P. E.

    2005-01-01

    Some pathogenesis-related genes are expressed in fungi only when the pathogen is in the host, but the host signals that trigger these gene expressions have not been identified. Virulent Nectria haematococca infects pea plants and requires either pelA, which is induced by pectin, or pelD, which is induced only in planta. However, the host signal(s) that trigger pelD expression was unknown. Here we report the isolation of the host signals and identify homoserine and asparagine, two free amino acids found in uniquely high levels in pea seedlings, as the pelD-inducing signals. N. haematococca has evolved a mechanism to sense the host tissue environment by using the high levels of two free amino acids in this plant, thereby triggering the expression of pelD to assist the pathogenic process. PMID:15753300

  16. Development of fiber optic spectroscopy for in-vitro and in-planta detection of fluorescent proteins

    NASA Astrophysics Data System (ADS)

    Liew, Oi Wah; Chen, Jun-Wei; Asundi, Anand K.

    2001-10-01

    The objective of this project is to apply photonics technology to bio-safety management of genetically modified (GM) plants. The conventional method for screening GM plants is through selection using antibiotic resistance markers. There is public concern with such approaches and these are associated with food safety issues, escape of antibiotic resistance genes to pathogenic microorganisms and interference with antibiotic therapy. Thus, the strategy taken in this project is to replace antibiotic resistance markers with fluorescent protein markers that allow for rapid and non-invasive optical screening of genetically modified plants. In this paper, fibre optic spectroscopy was developed to detect and quantify recombinant green (EGFP) and red (DsRED) fluorescent proteins in vitro and in planta. In vitro detection was first carried out to optimize the sensitivity of the optical system. The bacterial expression vectors carrying the coding regions of EGFP and DsRED were introduced into Escherichia coli host cells and fluorescent proteins were produced following induction with IPTG. Soluble EGFP and DsRED proteins were isolated from lysed bacterial cells and serially diluted for quantitative analysis by fibre optic spectroscopy using different light sources, namely, blue LED (475 nm), tungsten halogen (350 - 1000 nm) and double frequency Nd:YAG green laser (532 nm). Fluorescence near the expected emission wavelengths could be detected up to 320X dilution for EGFP and DsRED with blue LED and 532 nm green laser, respectively, as the excitation source. Tungsten halogen was found to be unsuitable for excitation of both EGFP and DsRED. EGFP was successfully purified by size separation under non-denaturing electrophoretic conditions and quantified. The minimum concentration of EGFP detectable with blue LED excitation was 5 mg/ml. To determine the capability of spectroscopy detection in planta, transgenic potato hairy roots and whole modified plant lines expressing the

  17. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance.

    PubMed

    Pandey, Sonika; Patel, Manish Kumar; Mishra, Avinash; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas.

  18. Phytohormonal networks promote differentiation of fiber initials on pre-anthesis cotton ovules grown in vitro and in planta.

    PubMed

    Kim, Hee Jin; Hinchliffe, Doug J; Triplett, Barbara A; Chen, Z Jeffrey; Stelly, David M; Yeater, Kathleen M; Moon, Hong S; Gilbert, Matthew K; Thyssen, Gregory N; Turley, Rickie B; Fang, David D

    2015-01-01

    The number of cotton (Gossypium sp.) ovule epidermal cells differentiating into fiber initials is an important factor affecting cotton yield and fiber quality. Despite extensive efforts in determining the molecular mechanisms regulating fiber initial differentiation, only a few genes responsible for fiber initial differentiation have been discovered. To identify putative genes directly involved in the fiber initiation process, we used a cotton ovule culture technique that controls the timing of fiber initial differentiation by exogenous phytohormone application in combination with comparative expression analyses between wild type and three fiberless mutants. The addition of exogenous auxin and gibberellins to pre-anthesis wild type ovules that did not have visible fiber initials increased the expression of genes affecting auxin, ethylene, ABA and jasmonic acid signaling pathways within 1 h after treatment. Most transcripts expressed differentially by the phytohormone treatment in vitro were also differentially expressed in the ovules of wild type and fiberless mutants that were grown in planta. In addition to MYB25-like, a gene that was previously shown to be associated with the differentiation of fiber initials, several other differentially expressed genes, including auxin/indole-3-acetic acid (AUX/IAA) involved in auxin signaling, ACC oxidase involved in ethylene biosynthesis, and abscisic acid (ABA) 8'-hydroxylase an enzyme that controls the rate of ABA catabolism, were co-regulated in the pre-anthesis ovules of both wild type and fiberless mutants. These results support the hypothesis that phytohormonal signaling networks regulate the temporal expression of genes responsible for differentiation of cotton fiber initials in vitro and in planta.

  19. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance

    PubMed Central

    Pandey, Sonika; Patel, Manish Kumar; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas. PMID:27411057

  20. The application of laser microdissection to in planta gene expression profiling of the maize anthracnose stalk rot fungus Colletotrichum graminicola.

    PubMed

    Tang, Weihua; Coughlan, Sean; Crane, Edmund; Beatty, Mary; Duvick, Jon

    2006-11-01

    Laser microdissection (LM) offers a potential means for deep sampling of a fungal plant-pathogen transcriptome during the infection process using whole-genome DNA microarrays. The use of a fluorescent protein-expressing fungus can greatly facilitate the identification of fungal structures for LM sampling. However, fixation methods that preserve both tissue histology and protein fluorescence, and that also yield RNA of suitable quality for microarray applications, have not been reported. We developed a microwave-accelerated acetone fixation, paraffin-embedding method that fulfills these requirements and used it to prepare mature maize stalk tissues infected with an Anemonia majano cyan fluorescent protein-expressing isolate of the anthracnose stalk rot fungus Colletotrichum graminicola. We successfully used LM to isolate individual maize cells associated with C. graminicola hyphae at an early stage of infection. The LM-derived RNA, after two-round linear amplification, was of sufficient quality and quantity for global expression profiling using a fungal microarray. Comparing replicated LM samples representing an early stage of stalk cell infection with samples from in vitro-germinated conidia, we identified 437 and 370 C. graminicola genes showing significant up- or downregulation, respectively. We confirmed the differential expression of several representative transcripts by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) and documented extensive overlap of this dataset with a PCR-subtraction library enriched for C. graminicola transcripts in planta. Our results demonstrate that LM is feasible for in planta pathogen expression profiling and can reveal clues about fungal genes involved in pathogenesis. The method in this report may be advantageous for visualizing a variety of cellular features that depend on a high degree of histochemical preservation and RNA integrity prior to LM.

  1. Phytohormonal Networks Promote Differentiation of Fiber Initials on Pre-Anthesis Cotton Ovules Grown In Vitro and In Planta

    PubMed Central

    Kim, Hee Jin; Hinchliffe, Doug J.; Triplett, Barbara A.; Chen, Z. Jeffrey; Stelly, David M.; Yeater, Kathleen M.; Moon, Hong S.; Gilbert, Matthew K.; Thyssen, Gregory N.; Turley, Rickie B.; Fang, David D.

    2015-01-01

    The number of cotton (Gossypium sp.) ovule epidermal cells differentiating into fiber initials is an important factor affecting cotton yield and fiber quality. Despite extensive efforts in determining the molecular mechanisms regulating fiber initial differentiation, only a few genes responsible for fiber initial differentiation have been discovered. To identify putative genes directly involved in the fiber initiation process, we used a cotton ovule culture technique that controls the timing of fiber initial differentiation by exogenous phytohormone application in combination with comparative expression analyses between wild type and three fiberless mutants. The addition of exogenous auxin and gibberellins to pre-anthesis wild type ovules that did not have visible fiber initials increased the expression of genes affecting auxin, ethylene, ABA and jasmonic acid signaling pathways within 1 h after treatment. Most transcripts expressed differentially by the phytohormone treatment in vitro were also differentially expressed in the ovules of wild type and fiberless mutants that were grown in planta. In addition to MYB25-like, a gene that was previously shown to be associated with the differentiation of fiber initials, several other differentially expressed genes, including auxin/indole-3-acetic acid (AUX/IAA) involved in auxin signaling, ACC oxidase involved in ethylene biosynthesis, and abscisic acid (ABA) 8'-hydroxylase an enzyme that controls the rate of ABA catabolism, were co-regulated in the pre-anthesis ovules of both wild type and fiberless mutants. These results support the hypothesis that phytohormonal signaling networks regulate the temporal expression of genes responsible for differentiation of cotton fiber initials in vitro and in planta. PMID:25927364

  2. Henrique da Rocha Lima.

    PubMed

    Bernardes Filho, Fred; Avelleira, João Carlos Regazzi

    2015-01-01

    Brazilian physician and researcher Henrique da Rocha Lima was born in 1879 in the city of Rio de Janeiro, where he studied medicine and obtained the degree of M.D. in 1901. He specialized in Clinical Medicine in Germany and was the ambassador in European countries of the scientific medicine that emerged from the Oswaldo Cruz Institute in the early twentieth century. Rocha Lima has discovered the causative agent of typhus and had a major contribution to the studies of yellow fever, Chagas disease, Carrión's disease and histoplasmosis. His genius, his research and his discoveries projected his name, and, with it, the image of Brazil in the international scientific scene.

  3. Berengario da Carpi.

    PubMed

    De Santo, N G; Bisaccia, C; De Santo, L S; De Santo, R M; Di Leo, V A; Papalia, T; Cirillo, M; Touwaide, A

    1999-01-01

    Berengario da Carpi was magister of anatomy and surgery at the University of Bologna from 1502 to 1527. Eustachio and Falloppia defined him as 'the restaurator of anatomy'. He was a great surgeon, anatomist and physician of illustrious patients including Lorenzo II dei Medici, Giovanni dalle Bande Nere, Galeazzo Pallavicini, Cardinal Colonna, and Alessandro Soderini. He had strong links to the intellectuals of his time (Forni, Bonamici, Manuzio, Pomponazzi) as well as with the Medici family. He was respected by the Popes Julius II, Leo X and Clement VII. His main contributions are the Isogogae Breves, De Fractura calvae sive cranei, and the illustrated Commentaria on the Anatomy of Mondino de Liucci, a textbook utilized for more than 200 years, which Berengario aimed to restore to its initial text. The Commentaria constitutes the material for the last part of this paper which concludes with a personal translation of some passages on 'The kidney', where the author gives poignant examples of experimental ingenuity.

  4. Henrique da Rocha Lima*

    PubMed Central

    Bernardes Filho, Fred; Avelleira, João Carlos Regazzi

    2015-01-01

    Brazilian physician and researcher Henrique da Rocha Lima was born in 1879 in the city of Rio de Janeiro, where he studied medicine and obtained the degree of M.D. in 1901. He specialized in Clinical Medicine in Germany and was the ambassador in European countries of the scientific medicine that emerged from the Oswaldo Cruz Institute in the early twentieth century. Rocha Lima has discovered the causative agent of typhus and had a major contribution to the studies of yellow fever, Chagas disease, Carrión’s disease and histoplasmosis. His genius, his research and his discoveries projected his name, and, with it, the image of Brazil in the international scientific scene. PMID:26131867

  5. The in planta transcriptome of Ralstonia solanacearum: conserved physiological and virulence strategies during bacterial wilt of tomato.

    PubMed

    Jacobs, Jonathan M; Babujee, Lavanya; Meng, Fanhong; Milling, Annett; Allen, Caitilyn

    2012-01-01

    Plant xylem fluid is considered a nutrient-poor environment, but the bacterial wilt pathogen Ralstonia solanacearum is well adapted to it, growing to 10(8) to 10(9) CFU/g tomato stem. To better understand how R. solanacearum succeeds in this habitat, we analyzed the transcriptomes of two phylogenetically distinct R. solanacearum strains that both wilt tomato, strains UW551 (phylotype II) and GMI1000 (phylotype I). We profiled bacterial gene expression at ~6 × 10(8) CFU/ml in culture or in plant xylem during early tomato bacterial wilt pathogenesis. Despite phylogenetic differences, these two strains expressed their 3,477 common orthologous genes in generally similar patterns, with about 12% of their transcriptomes significantly altered in planta versus in rich medium. Several primary metabolic pathways were highly expressed during pathogenesis. These pathways included sucrose uptake and catabolism, and components of these pathways were encoded by genes in the scrABY cluster. A UW551 scrA mutant was significantly reduced in virulence on resistant and susceptible tomato as well as on potato and the epidemiologically important weed host Solanum dulcamara. Functional scrA contributed to pathogen competitive fitness during colonization of tomato xylem, which contained ~300 µM sucrose. scrA expression was induced by sucrose, but to a much greater degree by growth in planta. Unexpectedly, 45% of the genes directly regulated by HrpB, the transcriptional activator of the type 3 secretion system (T3SS), were upregulated in planta at high cell densities. This result modifies a regulatory model based on bacterial behavior in culture, where this key virulence factor is repressed at high cell densities. The active transcription of these genes in wilting plants suggests that T3SS has a biological role throughout the disease cycle. IMPORTANCE Ralstonia solanacearum is a widespread plant pathogen that causes bacterial wilt disease. It inflicts serious crop losses on tropical

  6. "Plantas con madre": plants that teach and guide in the shamanic initiation process in the East-Central Peruvian Amazon.

    PubMed

    Jauregui, X; Clavo, Z M; Jovel, E M; Pardo-de-Santayana, M

    2011-04-12

    We present and discuss a particular group of plants used by a diversity of healers in the initiation process and apprenticeship of traditional medicine, as practiced by Amazonian societies in East-Central Peru. Often, these plants are locally called plantas con madre (plants with a mother), and are thought to guide initiates in the process of seeking sacred knowledge, learning about plant usage, and understanding traditional medicine practices. We illustrate the diversity of plants used in the apprenticeship and practice of traditional medicine, and nurture the discussion to better understand the terminology used by Indigenous healers to describe plant uses and their practices. The study was conducted between 2003 and 2008 with the participation of 29 curanderos (healers; 23 men, 6 women), 3 apprentices and 4 herbalists. The participants belonged to four ethnic groups: 17 Mestizos, 15 Shipibo-Konibo, 1 Ashaninka, and 1 Matsiguenga; a Spanish apprentice and an Italian herbalist were also included in the study. The field data were collected using semi-structured interviews, participant observation, and the witnessing of numerous healing sessions. Oral informed consent was obtained from each participant. We identified 55 plant species belonging to 26 botanical families, which are used in initiation processes and apprenticeships of traditional medicine. This group of plants is administered under strict conditions during training and healing sessions called dietas (shamanic diets), with the supervision of one or more maestros curanderos (master healers). We observed that during the shamanic diets, maestros curanderos administered plants depending on the teachings or tools he/she was passing on, and were based on a particular sequence during the initiation process: (I) purification and cleansing species; (II) sensitivity and intuition; (III) strengthening; and (IV) protection and defence. Traditional healers continue to be a primary source of health care for the majority

  7. WRS2 UPA DA Removal

    NASA Image and Video Library

    2009-11-23

    ISS021-E-032275 (23 Nov. 2009) --- NASA astronaut Leland Melvin, STS-129 mission specialist, holds the failed Urine Processor Assembly / Distillation Assembly (UPA DA) in the Destiny laboratory of the International Space Station while space shuttle Atlantis remains docked with the station. Melvin and European Space Agency astronaut Frank De Winne (out of frame), Expedition 21 commander, removed and packed the UPA DA, then transferred it from the Water Recovery System 2 (WRS-2) rack to Atlantis for stowage on the middeck.

  8. Detection of plum pox potyviral protein-protein interactions in planta using an optimized mRFP-based bimolecular fluorescence complementation system.

    PubMed

    Zilian, Eva; Maiss, Edgar

    2011-12-01

    In previous studies, protein interaction maps of different potyviruses have been generated using yeast two-hybrid (YTH) systems, and these maps have demonstrated a high diversity of interactions of potyviral proteins. Using an optimized bimolecular fluorescence complementation (BiFC) system, a complete interaction matrix for proteins of a potyvirus was developed for the first time under in planta conditions with ten proteins from plum pox virus (PPV). In total, 52 of 100 possible interactions were detected, including the self-interactions of CI, 6K2, VPg, NIa-Pro, NIb and CP, which is more interactions than have ever been detected for any other potyvirus in a YTH approach. Moreover, the BiFC system was shown to be able to localize the protein interactions, which was typified for the protein self-interactions indicated above. Additionally, experiments were carried out with the P3N-PIPO protein, revealing an interaction with CI but not with CP and supporting the involvement of P3N-PIPO in the cell-to-cell movement of potyviruses. No self-interaction of the PPV helper component-proteinase (HC-Pro) was detected using BiFC in planta. Therefore, additional experiments with turnip mosaic virus (TuMV) HC-Pro, PPV_HC-Pro and their mutants were conducted. The self-interaction of TuMV_HCpro, as recently demonstrated, and the self-interaction of the TuMV_ and PPV_HC-Pro mutants were shown by BiFC in planta, indicating that HC-Pro self-interactions may be species-specific. BiFC is a very useful and reliable method for the detection and localization of protein interactions in planta, thus enabling investigations under more natural conditions than studies in yeast cells.

  9. Closed-reference metatranscriptomics enables in planta profiling of putative virulence activities in the grapevine trunk-disease complex.

    PubMed

    Morales-Cruz, Abraham; Allenbeck, Gabrielle; Figueroa-Balderas, Rosa; Ashworth, Vanessa E; Lawrence, Daniel P; Travadon, Renaud; Smith, Rhonda J; Baumgartner, Kendra; Rolshausen, Philippe E; Cantu, Dario

    2017-02-20

    Grapevines, like other perennial crops, are affected by so-called 'trunk diseases', which damage the trunk and other woody tissues. Mature grapevines typically contract more than one trunk disease and often multiple grapevine trunk pathogens (GTPs) are recovered from infected tissues. The co-existence of different GTP species in complex and dynamic microbial communities complicates the study of the molecular mechanisms underlying disease development especially under vineyard conditions. The objective of this study was to develop and optimize a community-level transcriptomics (i.e., metatranscriptomics) approach that can monitor simultaneously the virulence activities of multiple GTPs in planta. The availability of annotated genomes for the most relevant co-infecting GTPs in diseased grapevine wood provided the unprecedented opportunity to generate a multi-species reference for mapping and quantifying DNA and RNA sequencing reads. We first evaluated popular sequence read mappers using permutations of multiple simulated datasets. Alignment parameters of the selected mapper were optimized to increase the specificity and sensitivity for its application to metagenomics and metatranscriptomics analyses. Initial testing on grapevine wood experimentally inoculated with individual GTPs confirmed the validity of the method. Using naturally-infected field samples expressing a variety of trunk disease symptoms, we show that our approach provides quantitative assessments of species composition as well as genome-wide transcriptional profiling of potential virulence factors, namely cell wall degradation, secondary metabolism and nutrient uptake for all co-infecting GTPs. This article is protected by copyright. All rights reserved.

  10. Agrobacterium tumefaciens deploys a superfamily of type VI secretion DNase effectors as weapons for interbacterial competition in planta.

    PubMed

    Ma, Lay-Sun; Hachani, Abderrahman; Lin, Jer-Sheng; Filloux, Alain; Lai, Erh-Min

    2014-07-09

    The type VI secretion system (T6SS) is a widespread molecular weapon deployed by many Proteobacteria to target effectors/toxins into both eukaryotic and prokaryotic cells. We report that Agrobacterium tumefaciens, a soil bacterium that triggers tumorigenesis in plants, produces a family of type VI DNase effectors (Tde) that are distinct from previously known polymorphic toxins and nucleases. Tde exhibits an antibacterial DNase activity that relies on a conserved HxxD motif and can be counteracted by a cognate immunity protein, Tdi. In vitro, A. tumefaciens T6SS could kill Escherichia coli but triggered a lethal counterattack by Pseudomonas aeruginosa upon injection of the Tde toxins. However, in an in planta coinfection assay, A. tumefaciens used Tde effectors to attack both siblings cells and P. aeruginosa to ultimately gain a competitive advantage. Such acquired T6SS-dependent fitness in vivo and conservation of Tde-Tdi couples in bacteria highlights a widespread antibacterial weapon beneficial for niche colonization. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  11. Cell wall targeted in planta iron accumulation enhances biomass conversion and seed iron concentration in Arabidopsis and rice

    DOE PAGES

    Yang, Haibing; Wei, Hui; Ma, Guojie; ...

    2016-04-07

    Conversion of nongrain biomass into liquid fuel is a sustainable approach to energy demands as global population increases. Previously, we showed that iron can act as a catalyst to enhance the degradation of lignocellulosic biomass for biofuel production. However, direct addition of iron catalysts to biomass pretreatment is diffusion-limited, would increase the cost and complexity of biorefinery unit operations and may have deleterious environmental impacts. Here, we show a new strategy for in planta accumulation of iron throughout the volume of the cell wall where iron acts as a catalyst in the deconstruction of lignocellulosic biomass. We engineered CBM-IBP fusionmore » polypeptides composed of a carbohydrate-binding module family 11 (CBM11) and an iron-binding peptide (IBP) for secretion into Arabidopsis and rice cell walls. CBM-IBP transformed Arabidopsis and rice plants show significant increases in iron accumulation and biomass conversion compared to respective controls. Further, CBM-IBP rice shows a 35% increase in seed iron concentration and a 40% increase in seed yield in greenhouse experiments. In conclusion, CBM-IBP rice potentially could be used to address iron deficiency, the most common and widespread nutritional disorder according to the World Health Organization.« less

  12. Cell wall targeted in planta iron accumulation enhances biomass conversion and seed iron concentration in Arabidopsis and rice.

    PubMed

    Yang, Haibing; Wei, Hui; Ma, Guojie; Antunes, Mauricio S; Vogt, Stefan; Cox, Joseph; Zhang, Xiao; Liu, Xiping; Bu, Lintao; Gleber, S Charlotte; Carpita, Nicholas C; Makowski, Lee; Himmel, Michael E; Tucker, Melvin P; McCann, Maureen C; Murphy, Angus S; Peer, Wendy A

    2016-10-01

    Conversion of nongrain biomass into liquid fuel is a sustainable approach to energy demands as global population increases. Previously, we showed that iron can act as a catalyst to enhance the degradation of lignocellulosic biomass for biofuel production. However, direct addition of iron catalysts to biomass pretreatment is diffusion-limited, would increase the cost and complexity of biorefinery unit operations and may have deleterious environmental impacts. Here, we show a new strategy for in planta accumulation of iron throughout the volume of the cell wall where iron acts as a catalyst in the deconstruction of lignocellulosic biomass. We engineered CBM-IBP fusion polypeptides composed of a carbohydrate-binding module family 11 (CBM11) and an iron-binding peptide (IBP) for secretion into Arabidopsis and rice cell walls. CBM-IBP transformed Arabidopsis and rice plants show significant increases in iron accumulation and biomass conversion compared to respective controls. Further, CBM-IBP rice shows a 35% increase in seed iron concentration and a 40% increase in seed yield in greenhouse experiments. CBM-IBP rice potentially could be used to address iron deficiency, the most common and widespread nutritional disorder according to the World Health Organization. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  13. Overexpression of the rice carotenoid cleavage dioxygenase 1 gene in Golden Rice endosperm suggests apocarotenoids as substrates in planta.

    PubMed

    Ilg, Andrea; Yu, Qiuju; Schaub, Patrick; Beyer, Peter; Al-Babili, Salim

    2010-08-01

    Carotenoids are converted by carotenoid cleavage dioxygenases that catalyze oxidative cleavage reactions leading to apocarotenoids. However, apocarotenoids can also be further truncated by some members of this enzyme family. The plant carotenoid cleavage dioxygenase 1 (CCD1) subfamily is known to degrade both carotenoids and apocarotenoids in vitro, leading to different volatile compounds. In this study, we investigated the impact of the rice CCD1 (OsCCD1) on the pigmentation of Golden Rice 2 (GR2), a genetically modified rice variety accumulating carotenoids in the endosperm. For this purpose, the corresponding cDNA was introduced into the rice genome under the control of an endosperm-specific promoter in sense and anti-sense orientations. Despite high expression levels of OsCCD1 in sense plants, pigment analysis revealed carotenoid levels and patterns comparable to those of GR2, pleading against carotenoids as substrates in rice endosperm. In support, similar carotenoid contents were determined in anti-sense plants. To check whether OsCCD1 overexpressed in GR2 endosperm is active, in vitro assays were performed with apocarotenoid substrates. HPLC analysis confirmed the cleavage activity of introduced OsCCD1. Our data indicate that apocarotenoids rather than carotenoids are the substrates of OsCCD1 in planta.

  14. A toolset of aequorin expression vectors for in planta studies of subcellular calcium concentrations in Arabidopsis thaliana

    PubMed Central

    Mehlmer, Norbert; Parvin, Nargis; Hurst, Charlotte H.; Knight, Marc R.; Teige, Markus; Vothknecht, Ute C.

    2014-01-01

    Calcium has long been acknowledged as one of the most important signalling components in plants. Many abiotic and biotic stimuli are transduced into a cellular response by temporal and spatial changes in cellular calcium concentration and the calcium-sensitive protein aequorin has been exploited as a genetically encoded calcium indicator for the measurement of calcium in planta. The objective of this work was to generate a compatible set of aequorin expression plasmids for the generation of transgenic plant lines to measure changes in calcium levels in different cellular subcompartments. Aequorin was fused to different targeting peptides or organellar proteins as a means to localize it to the cytosol, the nucleus, the plasma membrane, and the mitochondria. Furthermore, constructs were designed to localize aequorin in the stroma as well as the inner and outer surface of the chloroplast envelope membranes. The modular set-up of the plasmids also allows the easy replacement of targeting sequences to include other compartments. An additional YFP-fusion was included to verify the correct subcellular localization of all constructs by laser scanning confocal microscopy. For each construct, pBin19-based binary expression vectors driven by the 35S or UBI10 promoter were made for Agrobacterium-mediated transformation. Stable Arabidopsis lines were generated and initial tests of several lines confirmed their feasibility to measure calcium signals in vivo. PMID:22213817

  15. 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.

  16. In Planta Single-Molecule Pull-Down Reveals Tetrameric Stoichiometry of HD-ZIPIII:LITTLE ZIPPER Complexes[OPEN

    PubMed Central

    Husbands, Aman Y.; Aggarwal, Vasudha; Ha, Taekjip; Timmermans, Marja C.P.

    2016-01-01

    Deciphering complex biological processes markedly benefits from approaches that directly assess the underlying biomolecular interactions. Most commonly used approaches to monitor protein-protein interactions typically provide nonquantitative readouts that lack statistical power and do not yield information on the heterogeneity or stoichiometry of protein complexes. Single-molecule pull-down (SiMPull) uses single-molecule fluorescence detection to mitigate these disadvantages and can quantitatively interrogate interactions between proteins and other compounds, such as nucleic acids, small molecule ligands, and lipids. Here, we establish SiMPull in plants using the HOMEODOMAIN LEUCINE ZIPPER III (HD-ZIPIII) and LITTLE ZIPPER (ZPR) interaction as proof-of-principle. Colocalization analysis of fluorophore-tagged HD-ZIPIII and ZPR proteins provides strong statistical evidence of complex formation. In addition, we use SiMPull to directly quantify YFP and mCherry maturation probabilities, showing these differ substantially from values obtained in mammalian systems. Leveraging these probabilities, in conjunction with fluorophore photobleaching assays on over 2000 individual complexes, we determined HD-ZIPIII:ZPR stoichiometry. Intriguingly, these complexes appear as heterotetramers, comprising two HD-ZIPIII and two ZPR molecules, rather than heterodimers as described in the current model. This surprising result raises new questions about the regulation of these key developmental factors and is illustrative of the unique contribution SiMPull is poised to make to in planta protein interaction studies. PMID:27385814

  17. In Planta Single-Molecule Pull-Down Reveals Tetrameric Stoichiometry of HD-ZIPIII:LITTLE ZIPPER Complexes.

    PubMed

    Husbands, Aman Y; Aggarwal, Vasudha; Ha, Taekjip; Timmermans, Marja C P

    2016-08-01

    Deciphering complex biological processes markedly benefits from approaches that directly assess the underlying biomolecular interactions. Most commonly used approaches to monitor protein-protein interactions typically provide nonquantitative readouts that lack statistical power and do not yield information on the heterogeneity or stoichiometry of protein complexes. Single-molecule pull-down (SiMPull) uses single-molecule fluorescence detection to mitigate these disadvantages and can quantitatively interrogate interactions between proteins and other compounds, such as nucleic acids, small molecule ligands, and lipids. Here, we establish SiMPull in plants using the HOMEODOMAIN LEUCINE ZIPPER III (HD-ZIPIII) and LITTLE ZIPPER (ZPR) interaction as proof-of-principle. Colocalization analysis of fluorophore-tagged HD-ZIPIII and ZPR proteins provides strong statistical evidence of complex formation. In addition, we use SiMPull to directly quantify YFP and mCherry maturation probabilities, showing these differ substantially from values obtained in mammalian systems. Leveraging these probabilities, in conjunction with fluorophore photobleaching assays on over 2000 individual complexes, we determined HD-ZIPIII:ZPR stoichiometry. Intriguingly, these complexes appear as heterotetramers, comprising two HD-ZIPIII and two ZPR molecules, rather than heterodimers as described in the current model. This surprising result raises new questions about the regulation of these key developmental factors and is illustrative of the unique contribution SiMPull is poised to make to in planta protein interaction studies.

  18. Agrobacterium tumefaciens Deploys a Superfamily of Type VI Secretion DNase Effectors as Weapons for Interbacterial Competition In Planta

    PubMed Central

    Ma, Lay-Sun; Hachani, Abderrahman; Lin, Jer-Sheng; Filloux, Alain; Lai, Erh-Min

    2014-01-01

    Summary The type VI secretion system (T6SS) is a widespread molecular weapon deployed by many Proteobacteria to target effectors/toxins into both eukaryotic and prokaryotic cells. We report that Agrobacterium tumefaciens, a soil bacterium that triggers tumorigenesis in plants, produces a family of type VI DNase effectors (Tde) that are distinct from previously known polymorphic toxins and nucleases. Tde exhibits an antibacterial DNase activity that relies on a conserved HxxD motif and can be counteracted by a cognate immunity protein, Tdi. In vitro, A. tumefaciens T6SS could kill Escherichia coli but triggered a lethal counterattack by Pseudomonas aeruginosa upon injection of the Tde toxins. However, in an in planta coinfection assay, A. tumefaciens used Tde effectors to attack both siblings cells and P. aeruginosa to ultimately gain a competitive advantage. Such acquired T6SS-dependent fitness in vivo and conservation of Tde-Tdi couples in bacteria highlights a widespread antibacterial weapon beneficial for niche colonization. PMID:24981331

  19. 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

  20. Coronatine Gene Expression In Vitro and In Planta, and Protein Accumulation During Temperature Downshift in Pseudomonas syringae

    PubMed Central

    Braun, Yvonne; Smirnova, Angela V.; Weingart, Helge; Schenk, Alexander; Ullrich, Matthias S.

    2009-01-01

    The plant pathogenic bacterium Pseudomonas syringae PG4180 synthesizes high levels of the phytotoxin coronatine (COR) at the virulence-promoting temperature of 18 °C, but negligible amounts at 28 °C. Temperature-dependent COR gene expression is regulated by a modified two-component system, consisting of a response regulator, CorR, the histidine protein kinase CorS, and a third component, termed CorP. We analyzed at transcriptional and translational levels the expression of corS and the cma operon involved in COR biosynthesis after a temperature downshift from 28 to 18 °C. Expression of cma was induced within 20 min and increased steadily whereas corS expression was only slightly temperature-dependent. Accumulation of CmaB correlated with accumulation of cma mRNA. However, cma transcription was suppressed by inhibition of de novo protein biosynthesis. A transcriptional fusion of the cma promoter to a promoterless egfp gene was used to monitor the cma expression in vitro and in planta. A steady induction of cma::egfp by temperature downshift was observed in both environments. The results indicate that PG4180 responds to a temperature decrease with COR gene expression. However, COR gene expression and protein biosynthesis increased steadily, possibly reflecting adaptation to long-term rather than rapid temperature changes. PMID:22408526

  1. Construction of EGFP-labeling system for visualizing the infection process of Xanthomonas axonopodis pv. citri in planta.

    PubMed

    Liu, Li-Ping; Deng, Zi-Niu; Qu, Jin-Wang; Yan, Jia-Wen; Catara, Vittoria; Li, Da-Zhi; Long, Gui-You; Li, Na

    2012-09-01

    Xanthomonas axonopodis pv. citri (Xac) is the causal agent of citrus bacterial canker, an economically important disease to world citrus industry. To monitor the infection process of Xac in different citrus plants, the enhanced green florescent protein (EGFP) visualizing system was constructed to visualize the propagation and localization in planta. First, the wild-type Xac was isolated from the diseased leaves of susceptible 'Bingtang' sweet orange, and then the isolated Xac was labeled with EGFP by triparental mating. After PCR identification, the growth kinetics and pathogenicity of the transformants were analyzed in comparison with the wild-type Xac. The EGFP-labeled bacteria were inoculated by spraying on the surface and infiltration in the mesophyll of 'Bingtang' sweet orange leaves. The bacterial cell multiplication and diffusion processes were observed directly under confocal laser scanning microscope at different intervals after inoculation. The results indicated that the EGFP-labeled Xac releasing clear green fluorescence light under fluorescent microscope showed the infection process and had the same pathogenicity as the wild type to citrus. Consequently, the labeled Xac demonstrated the ability as an efficient tool to monitor the pathogen infection.

  2. Flavin Adenine Dinucleotide and N(5) ,N(10) -Methenyltetrahydrofolate are the in planta Cofactors of Arabidopsis thaliana Cryptochrome 3.

    PubMed

    Göbel, Tanja; Reisbacher, Stefan; Batschauer, Alfred; Pokorny, Richard

    2017-01-01

    Members of the cryptochrome/photolyase family (CPF) of proteins utilize noncovalently bound light-absorbing cofactors for their biological function. Usually, the identity of these cofactors is determined after expression in heterologous systems leaving the question unanswered whether these cofactors are identical to the indigenous ones. Here, cryptochrome 3 from Arabidopsis thaliana was expressed as a fusion with the green fluorescent protein in Arabidopsis plants. Besides the confirmation of the earlier report of its localization in chloroplasts, our data indicate that fractions of the fusion protein are present in the stroma and associated with thylakoids, respectively. Furthermore, it is shown that the fusion protein expressed in planta contains the same cofactors as the His6 -tagged protein expressed in Escherichia coli, that is, flavin adenine dinucleotide and N(5) ,N(10) -methenyltetrahydrofolate. This demonstrates that the heterologously expressed cryptochrome 3, characterized in a number of previous studies, is a valid surrogate of the corresponding protein expressed in plants. To our knowledge, this is also a first conclusive analysis of cofactors bound to an Arabidopsis protein belonging to the CPF and purified from plant tissue. © 2016 The Authors. Photochemistry and Photobiology published by Wiley Periodicals, Inc. on behalf of American Society for Photobiology.

  3. In planta chemical cross-linking and mass spectrometry analysis of protein structure and interaction in Arabidopsis.

    PubMed

    Zhu, Xinliang; Yu, Fengchao; Yang, Zhu; Liu, Shichang; Dai, Chen; Lu, Xiaoyun; Liu, Chenyu; Yu, Weichuan; Li, Ning

    2016-07-01

    Site-specific chemical cross-linking in combination with mass spectrometry analysis has emerged as a powerful proteomic approach for studying the three-dimensional structure of protein complexes and in mapping protein-protein interactions (PPIs). Building on the success of MS analysis of in vitro cross-linked proteins, which has been widely used to investigate specific interactions of bait proteins and their targets in various organisms, we report a workflow for in vivo chemical cross-linking and MS analysis in a multicellular eukaryote. This approach optimizes the in vivo protein cross-linking conditions in Arabidopsis thaliana, establishes a MudPIT procedure for the enrichment of cross-linked peptides, and develops an integrated software program, exhaustive cross-linked peptides identification tool (ECL), to identify the MS spectra of in planta chemical cross-linked peptides. In total, two pairs of in vivo cross-linked peptides of high confidence have been identified from two independent biological replicates. This work demarks the beginning of an alternative proteomic approach in the study of in vivo protein tertiary structure and PPIs in multicellular eukaryotes. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Sucrose phosphate synthase and sucrose phosphate phosphatase interact in planta and promote plant growth and biomass accumulation.

    PubMed

    Maloney, Victoria J; Park, Ji-Young; Unda, Faride; Mansfield, Shawn D

    2015-07-01

    Bioinformatic analysis indicates that sucrose phosphate synthase (SPS) contains a putative C-terminal sucrose phosphate phosphatase (SPP)-like domain that may facilitates the binding of SPP. If an SPS-SPP enzyme complex exists, it may provide sucrose biosynthesis with an additional level of regulation, forming a direct metabolic channel for sucrose-6-phosphate between these two enzymes. Herein, the formation of an enzyme complex between SPS and SPP was examined, and the results from yeast two-hybrid experiments suggest that there is indeed an association between these proteins. In addition, in planta bioluminescence resonance energy transfer (BRET) was observed in Arabidopsis seedlings, providing physical evidence for a protein interaction in live cells and in real time. Finally, bimolecular fluorescence complementation (BiFC) was employed in an attempt to detect SPS-SPP interactions visually. The findings clearly demonstrated that SPS interacts with SPP and that this interaction impacts soluble carbohydrate pools and affects carbon partitioning to starch. Moreover, a fusion construct between the two genes promotes plant growth in both transgenic Arabidopsis and hybrid poplar. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  5. Biochemical synthesis of uniformly (13)C-labeled diterpene hydrocarbons and their bioconversion to diterpenoid phytoalexins in planta.

    PubMed

    Ye, Zhongfeng; Nakagawa, Kazuya; Natsume, Masahiro; Nojiri, Hideaki; Kawaide, Hiroshi; Okada, Kazunori

    2017-06-01

    Phytocassanes and momilactones are the major diterpenoid phytoalexins inductively produced in rice as bioactive substances. Regardless of extensive studies on the biosynthetic pathways of these phytoalexins, bioconversion of diterpene hydrocarbons is not shown in planta. To elucidate the entire biosynthetic pathways of these phytoalexins, uniformly (13)C-labeled ent-cassadiene and syn-pimaradiene were enzymatically synthesized with structural verification by GC-MS and (13)C-NMR. Application of the (13)C-labeled substrates on rice leaves led to the detection of (13)C-labeled metabolites using LC-MS/MS. Further application of this method in the moss Hypnum plumaeforme and the nearest out-group of Oryza species Leersia perrieri, respectively, resulted in successful bioconversion of these labeled substrates into phytoalexins in these plants. These results demonstrate that genuine biosynthetic pathways from these diterpene hydrocarbons to the end product phytoalexins occur in these plants and that enzymatically synthesized [U-(13)C20] diterpene substrates are a powerful tool for chasing endogenous metabolites without dilution with naturally abundant unlabeled compounds.

  6. In planta transformation of sorghum (Sorghum bicolor (L.) Moench) using TPS1 gene for enhancing tolerance to abiotic stresses.

    PubMed

    Yellisetty, Varalaxmi; Reddy, L A; Mandapaka, Maheswari

    2015-09-01

    An in planta transformation protocol for sorghum (Sorghum bicolor (L.) Moench) using shoot apical meristem of germinating seedlings is reported in this study. Agrobacterium tumefaciens strain, LBA4404 with pCAMBIA1303 vector and construct pCAMBIA1303TPS1 were individually used for transformation. Since, the transgene is integrated into the cells of already differentiated tissues, the T 0 plants were chimeric and stable integration was observed in T1 generation. β-Glucuronidase (GUS) expression in the seedlings and spikelets of emerging cob was the first indication of transformability in T0 generation which was further confirmed by PCR analysis using hpt and TPS1 gene-specific primers. Screening on 25 mg/L hygromycin combined with PCR analysis was used for selection of transformants in the T1 generation. Transformation efficiencies ranged between 34-38% and 26-34% using pCAMBIA1303 vector and construct pCAMBIA1303TPS1, respectively. Molecular characterization of the T2 transgenics using PCR, RT-PCR and Southern blot analyses further revealed the integration, expression and inheritance of the transgene. These results indicate the feasibility of the method to generate transgenics with pCAM-BIA1303 vector and construct pCAMBIA1303TPS1. The abiotic stress tolerance of TPS1 transgenics developed in the present study was evident by the ability of the transformants to tolerate 200 mM NaCl as well as higher root growth and biomass.

  7. In planta distribution of 'Candidatus Liberibacter asiaticus' as revealed by polymerase chain reaction (PCR) and real-time PCR.

    PubMed

    Tatineni, Satyanarayana; Sagaram, Uma Shankar; Gowda, Siddarame; Robertson, Cecile J; Dawson, William O; Iwanami, Toru; Wang, Nian

    2008-05-01

    Huanglongbing (HLB) is one of the most devastating diseases of citrus worldwide, and is caused by a phloem-limited fastidious prokaryotic alpha-proteobacterium that is yet to be cultured. In this study, a combination of traditional polymerase chain reaction (PCR) and real-time PCR targeting the putative DNA polymerase and 16S rDNA sequence of 'Candidatus Liberibacter asiaticus,' respectively, were used to examine the distribution and movement of the HLB pathogen in the infected citrus tree. We found that 'Ca. Liberibacter asiaticus' was distributed in bark tissue, leaf midrib, roots, and different floral and fruit parts, but not in endosperm and embryo, of infected citrus trees. Quantification analysis of the HLB bacterium indicated that it was distributed unevenly in planta and ranged from 14 to 137,031 cells/mug of total DNA in different tissues. A relatively high concentration of 'Ca. Liberibacter asiaticus' was observed in fruit peduncles. Our data from greenhouse-infected plants also indicated that 'Ca. Liberibacter asiaticus' was transmitted systemically from infection site to different parts of the plant. Understanding the distribution and movement of the HLB bacterium inside an individual citrus tree is critical for discerning its virulence mechanism and to develop management strategies for HLB.

  8. Functional characterisation of metal(loid) processes in planta through the integration of synchrotron techniques and plant molecular biology

    PubMed Central

    Donner, Erica; Punshon, Tracy; Guerinot, Mary Lou; Lombi, Enzo

    2013-01-01

    Functional characterisation of the genes regulating metal(loid) homeostasis in plants is a major focus of crop biofortification, phytoremediation, and food security research. This paper focuses on the potential for advancing plant metal(loid) research by combining molecular biology and synchrotron-based techniques. Recent advances in x-ray focussing optics and fluorescence detection have greatly improved the potential of synchrotron techniques for plant science research, allowing metal(loids) to be imaged in vivo in hydrated plant tissues at sub-micron resolution. Laterally resolved metal(loid) speciation can also be determined. By using molecular techniques to probe the location of gene expression and protein localisation and combining it with this synchrotron-derived data, functional information can be effectively and efficiently assigned to specific genes. This paper provides a review of the state of the art in this field, and provides examples as to how synchrotron-based methods can be combined with molecular techniques to facilitate functional characterisation of genes in planta. PMID:22200921

  9. Validation of RT-qPCR reference genes for in planta expression studies in Hemileia vastatrix, the causal agent of coffee leaf rust.

    PubMed

    Vieira, Ana; Talhinhas, Pedro; Loureiro, Andreia; Duplessis, Sébastien; Fernandez, Diana; Silva, Maria do Céu; Paulo, Octávio S; Azinheira, Helena Gil

    2011-09-01

    Hemileia vastatrix is a biotrophic fungus, causing coffee leaf rust in all coffee growing countries, leading to serious social and economic problems. Gene expression studies may have a key role unravelling the transcriptomics of this pathogen during interaction with the plant host. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is currently the golden standard for gene expression analysis, although an accurate normalisation is essential for adequate conclusions. Reference genes are often used for this purpose, but the stability of their expression levels requires validation under experimental conditions. Moreover, pathogenic fungi undergo important biomass variations along their infection process in planta, which raises the need for an adequate method to further normalise the proportion of fungal cDNA in the total plant and fungus cDNA pool. In this work, the expression profiles of seven reference genes [glyceraldehyde-3-phosphate dehydrogenase (GADPH), elongation factor (EF-1), Beta tubulin (β-tubulin), cytochrome c oxidase subunit III (Cyt III), cytochrome b (Cyt b), Hv00099, and 40S ribosomal protein (40S_Rib)] were analysed across 28 samples, obtained in vitro (germinated uredospores and appressoria) and in planta (post-penetration fungal growth phases). Gene stability was assessed using the statistical algorithms incorporated in geNorm and NormFinder tools. Cyt b, 40S_Rib, and Hv00099 were the most stable genes for the in vitro dataset, while 40S_Rib, GADPH, and Cyt III were the most stable in planta. For the combined datasets (in vitro and in planta), 40S_Rib, GADPH, and Hv00099 were selected as the most stable. Subsequent expression analysis for a gene encoding an alpha subunit of a heterotrimeric G-protein showed that the reference genes selected for the combined dataset do not differ significantly from those selected specifically for the in vitro and in planta datasets. Our study provides tools for correct validation

  10. Effective and specific in planta RNAi in cyst nematodes: expression interference of four parasitism genes reduces parasitic success.

    PubMed

    Sindhu, Anoop S; Maier, Tom R; Mitchum, Melissa G; Hussey, Richard S; Davis, Eric L; Baum, Thomas J

    2009-01-01

    Cyst nematodes are highly evolved sedentary plant endoparasites that use parasitism proteins injected through the stylet into host tissues to successfully parasitize plants. These secretory proteins likely are essential for parasitism as they are involved in a variety of parasitic events leading to the establishment of specialized feeding cells required by the nematode to obtain nourishment. With the advent of RNA interference (RNAi) technology and the demonstration of host-induced gene silencing in parasites, a new strategy to control pests and pathogens has become available, particularly in root-knot nematodes. Plant host-induced silencing of cyst nematode genes so far has had only limited success but similarly should disrupt the parasitic cycle and render the host plant resistant. Additional in planta RNAi data for cyst nematodes are being provided by targeting four parasitism genes through host-induced RNAi gene silencing in transgenic Arabidopsis thaliana, which is a host for the sugar beet cyst nematode Heterodera schachtii. Here it is reported that mRNA abundances of targeted nematode genes were specifically reduced in nematodes feeding on plants expressing corresponding RNAi constructs. Furthermore, this host-induced RNAi of all four nematode parasitism genes led to a reduction in the number of mature nematode females. Although no complete resistance was observed, the reduction of developing females ranged from 23% to 64% in different RNAi lines. These observations demonstrate the relevance of the targeted parasitism genes during the nematode life cycle and, potentially more importantly, suggest that a viable level of resistance in crop plants may be accomplished in the future using this technology against cyst nematodes.

  11. Efficient In Planta Detection and Dissection of De Novo Mutation Events in the Arabidopsis thaliana Disease Resistance Gene UNI.

    PubMed

    Ogawa, Tomohiko; Mori, Akiko; Igari, Kadunari; Morita, Miyo Terao; Tasaka, Masao; Uchida, Naoyuki

    2016-06-01

    Plants possess disease resistance (R) proteins encoded by R genes, and each R protein recognizes a specific pathogen factor(s) for immunity. Interestingly, a remarkably high degree of polymorphisms in R genes, which are traces of past mutation events during evolution, suggest the rapid diversification of R genes. However, little is known about molecular aspects that facilitate the rapid change of R genes because of the lack of tools that enable us to monitor de novo R gene mutations efficiently in an experimentally feasible time scale, especially in living plants. Here we introduce a model assay system that enables efficient in planta detection of de novo mutation events in the Arabidopsis thaliana R gene UNI in one generation. The uni-1D mutant harbors a gain-of-function allele of the UNI gene. uni-1D heterozygous individuals originally exhibit dwarfism with abnormally short stems. However, interestingly, morphologically normal stems sometimes emerge spontaneously from the uni-1D plants, and the morphologically reverted tissues carry additional de novo mutations in the UNI gene. Strikingly, under an extreme condition, almost half of the examined population shows the reversion phenomenon. By taking advantage of this phenomenon, we demonstrate that the reversion frequency is remarkably sensitive to a variety of fluctuations in DNA stability, underlying a mutable tendency of the UNI gene. We also reveal that activities of the salicylic acid pathway and DNA damage sensor pathway are involved in the reversion phenomenon. Thus, we provide an experimentally feasible model tool to explore factors and conditions that significantly affect the R gene mutation phenomenon. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  12. Identification of Epichloë Endophytes In Planta by a Microsatellite-Based PCR Fingerprinting Assay with Automated Analysis

    PubMed Central

    Moon, Christina D.; Tapper, Brian A.; Scott, Barry

    1999-01-01

    Epichloë endophytes are a group of filamentous fungi that include both sexual (Epichloë) and asexual (Neotyphodium) species. As a group they are genetically diverse and form both antagonistic and mutualistic associations with temperate grasses. We report here on the development of a microsatellite-based PCR system for fingerprinting this group of fungi with template isolated from either culture or infected plant material. M13mp19 partial genomic libraries were constructed for size-fractionated genomic DNA from two endophyte strains. These libraries were screened with a mixture of DIG-labeled dinucleotide and trinucleotide repeat probes. Positive clones were sequenced, and nine unique microsatellite loci were identified. An additional microsatellite was serendipitously identified in the 3′ untranscribed region of the 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase gene from N. lolii Lp19. Primers were designed for each locus and a panel of endophytes, from different taxonomic groupings, was screened to determine the degree of polymorphism. On the basis of these results a multiplex assay was developed for strain identification with fluorescently labeled primers for five of these loci. Using this system the size of the products amplified can be precisely determined by automated analysis, and an allele profile for each strain can be readily generated. The assay was shown to resolve endophyte groupings to the level of known isozyme phenotype groupings. In a blind test the assay was used successfully to identify a set of endophytes in planta. A reference database of allele sizes has been established for the panel of endophytes examined, and this will be expanded as new strains are analyzed. PMID:10049893

  13. Expression of the Phytophthora infestans ipiB and ipiO genes in planta and in vitro.

    PubMed

    Pieterse, C M; Derksen, A M; Folders, J; Govers, F

    1994-08-02

    The ipiB and ipiO genes of the potato late blight fungus Phytophthora infestans (Mont.) de Bary were isolated from a genomic library in a screen for genes induced in planta. Expression of these genes was studied during pathogenesis on various host tissues and different host plants, some of which show specific resistance against P. infestans infection. During pathogenesis on leaves and tubers of the fully susceptible potato cultivar (cv.) Ajax and on leaves of the fully susceptible tomato cv. Moneymaker, the P. infestans ipiB and ipiO genes show a transient expression pattern with highest mRNA levels in the early stages of infection. During the interaction with leaves of the partially resistant potato cv. Pimpernel, the expression is also transient but accumulation and disappearance of the mRNAs is delayed. Also in P. infestans inoculated onto a race-specific resistant potato cultivar and onto the nonhost Solanum nigrum, ipiB and ipiO mRNA is detectable during the initial stages of infection. Apparently, the expression of the ipiB and the ipiO genes is activated in compatible, incompatible and nonhost interactions. In encysted zoospores, ipiB and ipiO mRNA accumulation was not detectable, but during cyst germination and appressorium formation on an artificial surface the genes are highly expressed. Expression studies in mycelium grown in vitro revealed that during nutrient starvation the expression of the ipiB and ipiO genes is induced. For ipiO gene expression, carbon deprivation appeared to be sufficient. The ipiO gene promoters contain a sequence motif that functions as a glucose repression element in yeast and this motif might be involved in the regulation of ipiO gene expression.

  14. In planta AKT2 subunits constitute a pH- and Ca2+-sensitive inward rectifying K+ channel.

    PubMed

    Latz, Andreas; Ivashikina, Natalya; Fischer, Susanne; Ache, Peter; Sano, Toshio; Becker, Dirk; Deeken, Rosalia; Hedrich, Rainer

    2007-04-01

    Heterologous expression of plant genes in yeast and animal cells represents a common approach to study plant ion channels. When expressed in Xenopus oocytes and COS cells the Arabidopsis Shaker-like K+ channel, AKT2 forms a weakly voltage-dependent channel, blocked by Ca2+ and protons. Channels with these characteristics, however, were not found in AKT2-expressing Arabidopsis cell types. To understand this phenomenon, we employed Agrobacterium-mediated transient transformation to functionally characterise Arabidopsis thaliana channels in Nicotiana benthamiana mesophyll cells. In this expression system we used AtTPK4 as a control for voltage-independent A. thaliana channels. Agrobacteria harbouring GFP-tagged constructs with the coding sequences of AKT2 and AtTPK4 were infiltrated into intact tobacco leaves. With quantitative RT-PCR analyses channel transcripts of AKT2 and AtTPK4 were determined in transformed leaves. These results were confirmed by Western blots with V5 epitope-tagged AKT2 and AtTPK4 proteins, showing that the channel protein was indeed synthesised. For functional analysis of these channels, mesophyll protoplasts were isolated from infiltrated leaf sections. Patch-clamp studies revealed that AKT2 channels in mesophyll protoplasts retained Ca2+ and pH sensitivity, characteristics of the heterologously expressed protein, but displayed pronounced differences in voltage-dependence and kinetics. AKT2-transformed mesophyll cells, displayed inward-rectifying, rather than voltage-independent K+ channels, initially characterised in AKT2-expressing animal cells. In contrast, AtTPK4 showed the same electrophysiological characteristics both, in oocytes and plant cells. Our data suggest that heterologous systems do not always possess all regulatory components for functional expression of plant channels. Therefore, transient expression of plant proteins in planta provides an additional research tool for rapid biophysical analysis of plant ion channels.

  15. Barcoding the kingdom Plantae: new PCR primers for ITS regions of plants with improved universality and specificity.

    PubMed

    Cheng, Tao; Xu, Chao; Lei, Li; Li, Changhao; Zhang, Yu; Zhou, Shiliang

    2016-01-01

    The internal transcribed spacer (ITS) of nuclear ribosomal DNA is one of the most commonly used DNA markers in plant phylogenetic and DNA barcoding analyses, and it has been recommended as a core plant DNA barcode. Despite this popularity, the universality and specificity of PCR primers for the ITS region are not satisfactory, resulting in amplification and sequencing difficulties. By thoroughly surveying and analysing the 18S, 5.8S and 26S sequences of Plantae and Fungi from GenBank, we designed new universal and plant-specific PCR primers for amplifying the whole ITS region and a part of it (ITS1 or ITS2) of plants. In silico analyses of the new and the existing ITS primers based on these highly representative data sets indicated that (i) the newly designed universal primers are suitable for over 95% of plants in most groups; and (ii) the plant-specific primers are suitable for over 85% of plants in most groups without amplification of fungi. A total of 335 samples from 219 angiosperm families, 11 gymnosperm families, 24 fern and lycophyte families, 16 moss families and 17 fungus families were used to test the performances of these primers. In vitro PCR produced similar results to those from the in silico analyses. Our new primer pairs gave PCR improvements up to 30% compared with common-used ones. The new universal ITS primers will find wide application in both plant and fungal biology, and the new plant-specific ITS primers will, by eliminating PCR amplification of nonplant templates, significantly improve the quality of ITS sequence information collections in plant molecular systematics and DNA barcoding.

  16. A Real-Time Fluorogenic Assay for the Visualization of Glycoside Hydrolase Activity in Planta1[C][OA

    PubMed Central

    Ibatullin, Farid M.; Banasiak, Alicja; Baumann, Martin J.; Greffe, Lionel; Takahashi, Junko; Mellerowicz, Ewa J.; Brumer, Harry

    2009-01-01

    There currently exists a diverse array of molecular probes for the in situ localization of polysaccharides, nucleic acids, and proteins in plant cells, including reporter enzyme strategies (e.g. protein-glucuronidase fusions). In contrast, however, there is a paucity of methods for the direct analysis of endogenous glycoside hydrolases and transglycosidases responsible for cell wall remodeling. To exemplify the potential of fluorogenic resorufin glycosides to address this issue, a resorufin β-glycoside of a xylogluco-oligosaccharide (XXXG-β-Res) was synthesized as a specific substrate for in planta analysis of XEH activity. The resorufin aglycone is particularly distinguished for high sensitivity in muro assays due to a low pKa (5.8) and large extinction coefficient (ε 62,000 m−1cm−1), long-wavelength fluorescence (excitation 571 nm/emission 585 nm), and high quantum yield (0.74) of the corresponding anion. In vitro analyses demonstrated that XXXG-β-Res is hydrolyzed by the archetypal plant XEH, nasturtium (Tropaeolum majus) NXG1, with classical Michaelis-Menten substrate saturation kinetics and a linear dependence on both enzyme concentration and incubation time. Further, XEH activity could be visualized in real time by observing the localized increase in fluorescence in germinating nasturtium seeds and Arabidopsis (Arabidopsis thaliana) inflorescent stems by confocal microscopy. Importantly, this new in situ XEH assay provides an essential complement to the in situ xyloglucan endotransglycosylase assay, thus allowing delineation of the disparate activities encoded by xyloglucan endotransglycosylase/hydrolase genes directly in plant tissues. The observation that XXXG-β-Res is also hydrolyzed by diverse microbial XEHs indicates that this substrate, and resorufin glycosides in general, may find broad applicability for the analysis of wall restructuring by polysaccharide hydrolases during morphogenesis and plant-microbe interactions. PMID:19783642

  17. WRS2 UPA DA Removal

    NASA Image and Video Library

    2009-11-23

    ISS021-E-032273 (23 Nov. 2009) --- European Space Agency astronaut Frank De Winne, Expedition 21 commander, holds the failed Urine Processor Assembly / Distillation Assembly (UPA DA) in the Destiny laboratory of the International Space Station while space shuttle Atlantis remains docked with the station. De Winne and NASA astronaut Leland Melvin (out of frame), STS-129 mission specialist, removed and packed the UPA DA, then transferred it from the Water Recovery System 2 (WRS-2) rack to Atlantis for stowage on the middeck.

  18. da Vinci decoded: does da Vinci stereopsis rely on disparity?

    PubMed

    Tsirlin, Inna; Wilcox, Laurie M; Allison, Robert S

    2012-11-01

    In conventional stereopsis, the depth between two objects is computed based on the retinal disparity in the position of matching points in the two eyes. When an object is occluded by another object in the scene, so that it is visible only in one eye, its retinal disparity cannot be computed. Nakayama and Shimojo (1990) found that a precept of quantitative depth between the two objects could still be established for such stimuli and proposed that this precept is based on the constraints imposed by occlusion geometry. They named this and other occlusion-based depth phenomena "da Vinci stereopsis." Subsequent research found quantitative depth based on occlusion geometry in several other classes of stimuli grouped under the term da Vinci stereopsis. However, Nakayama and Shimojo's findings were later brought into question by Gillam, Cook, and Blackburn (2003), who suggested that quantitative depth in their stimuli was perceived based on conventional disparity. In order to understand whether da Vinci stereopsis relies on one type of mechanism or whether its function is stimulus dependent we examine the nature and source of depth in the class of stimuli used by Nakayama and Shimojo (1990). We use three different psychophysical and computational methods to show that the most likely source for depth in these stimuli is occlusion geometry. Based on these experiments and previous data we discuss the potential mechanisms responsible for processing depth from monocular features in da Vinci stereopsis.

  19. Leonardo da Vinci and the Downburst.

    NASA Astrophysics Data System (ADS)

    Gedzelman, Stanley David

    1990-05-01

    Evidence from the drawings, experiments, and writings of Leonardo da Vinci are presented to demonstrate that da Vinci recognized and, possibly, discovered the downburst and understood its associated airflow. Other early references to vortex flows resembling downbursts are mentioned.

  20. Bikinin-like inhibitors targeting GSK3/Shaggy-like kinases: characterisation of novel compounds and elucidation of their catabolism in planta

    PubMed Central

    2014-01-01

    Background Plant GSK-3/Shaggy-like kinases are key players in brassinosteroid (BR) signalling which impact on plant development and participate in response to wounding, pathogens and salt stress. Bikinin was previously identified in a chemical genetics screen as an inhibitor targeting these kinases. To dissect the structural elements crucial for inhibition of GSK-3/Shaggy-like kinases by bikinin and to isolate more potent compounds we synthesised a number of related substances and tested their inhibitory activity in vitro and in vivo using Arabidopsis thaliana. Results A pyridine ring with an amido succinic acid residue in position 2 and a halogen in position 5 were crucial for inhibitory activity. The compound with an iodine substituent in position 5, denoted iodobikinin, was most active in inhibiting BIN2 activity in vitro and efficiently induced brassinosteroid-like responses in vivo. Its methyl ester, methyliodobikinin, showed improved cell permeability, making it highly potent in vivo although it had lower activity in vitro. HPLC analysis revealed that the methyl residue was rapidly cleaved off in planta liberating active iodobikinin. In addition, we provide evidence that iodobikinin and bikinin are inactivated in planta by conjugation with glutamic acid or malic acid and that the latter process is catalysed by the malate transferase SNG1. Conclusion Brassinosteroids participate in regulation of many aspects of plant development and in responses to environmental cues. Thus compounds modulating their action are valuable tools to study such processes and may be an interesting opportunity to modify plant growth and performance in horticulture and agronomy. Here we report the development of bikinin derivatives with increased potency that can activate BR signalling and mimic BR action. Methyliodobikinin was 3.4 times more active in vivo than bikinin. The main reason for the superior activity of methyliodobikinin, the most potent compound, is its enhanced plant

  1. Requirement of the galU Gene for Polysaccharide Production by and Pathogenicity and Growth In Planta of Xanthomonas citri subsp. citri▿

    PubMed Central

    Guo, Yinping; Sagaram, Uma Shankar; Kim, Jeong-soon; Wang, Nian

    2010-01-01

    Xanthomonas citri subsp. citri is the causal agent of citrus canker, which has a significant impact on citrus production. In this study, we characterized the galU gene of X. citri subsp. citri. Two galU mutants (F6 and D12) were identified in an X. citri subsp. citri EZ-Tn5 Tnp transposon library. Rescue cloning, sequence analysis, and Southern blot analysis indicated that both of these mutants had a single copy of the EZ-Tn5 transposon inserted in galU in the chromosome. Further study showed that galU was required for biosynthesis of extracellular polysaccharides (EPS; xanthan gum) and capsular polysaccharide (CPS) and biofilm formation. Mutation of galU resulted in a loss of pathogenicity for grapefruit. The loss of pathogenicity of a galU mutant resulted from its inability to grow in planta rather than from the effect on virulence genes. Quantitative reverse transcription-PCR assays indicated that mutation of galU did not impair the expression of key virulence genes, such as pthA of X. citri subsp. citri. Although D12 had a growth rate similar to that of the wild-type strain in nutrient broth, no D12 population became established in the intercellular spaces of citrus leaves. Coinoculation of a galU mutant with the wild-type strain did not promote growth of the galU mutant in planta. Defects in EPS and CPS production, pathogenicity, and growth in planta of the galU mutant were complemented to the wild-type level using plasmid pCGU2.1 containing an intact galU gene. These data indicate that the galU gene contributes to X. citri subsp. citri growth in intercellular spaces and is involved in EPS and CPS synthesis and biofilm formation. PMID:20118360

  2. Bikinin-like inhibitors targeting GSK3/Shaggy-like kinases: characterisation of novel compounds and elucidation of their catabolism in planta.

    PubMed

    Rozhon, Wilfried; Wang, Wuyan; Berthiller, Franz; Mayerhofer, Juliane; Chen, Tingting; Petutschnig, Elena; Sieberer, Tobias; Poppenberger, Brigitte; Jonak, Claudia

    2014-06-19

    Plant GSK-3/Shaggy-like kinases are key players in brassinosteroid (BR) signalling which impact on plant development and participate in response to wounding, pathogens and salt stress. Bikinin was previously identified in a chemical genetics screen as an inhibitor targeting these kinases. To dissect the structural elements crucial for inhibition of GSK-3/Shaggy-like kinases by bikinin and to isolate more potent compounds we synthesised a number of related substances and tested their inhibitory activity in vitro and in vivo using Arabidopsis thaliana. A pyridine ring with an amido succinic acid residue in position 2 and a halogen in position 5 were crucial for inhibitory activity. The compound with an iodine substituent in position 5, denoted iodobikinin, was most active in inhibiting BIN2 activity in vitro and efficiently induced brassinosteroid-like responses in vivo. Its methyl ester, methyliodobikinin, showed improved cell permeability, making it highly potent in vivo although it had lower activity in vitro. HPLC analysis revealed that the methyl residue was rapidly cleaved off in planta liberating active iodobikinin. In addition, we provide evidence that iodobikinin and bikinin are inactivated in planta by conjugation with glutamic acid or malic acid and that the latter process is catalysed by the malate transferase SNG1. Brassinosteroids participate in regulation of many aspects of plant development and in responses to environmental cues. Thus compounds modulating their action are valuable tools to study such processes and may be an interesting opportunity to modify plant growth and performance in horticulture and agronomy. Here we report the development of bikinin derivatives with increased potency that can activate BR signalling and mimic BR action. Methyliodobikinin was 3.4 times more active in vivo than bikinin. The main reason for the superior activity of methyliodobikinin, the most potent compound, is its enhanced plant tissue permeability. Inactivation

  3. Estudo exaustivo e sistemático de erros nas provas da olimpíada brasileira de astronomia - resultados preliminares

    NASA Astrophysics Data System (ADS)

    Oliveira, F. S.; Canalle, J. B. G.; Villas da Rocha, J. F.

    2003-08-01

    A Olimpíada Brasileira de Astronomia (OBA) contou, em sua quinta edição, com a participação de mais de 60 mil alunos dos ensinos fundamental e médio de praticamente todo o território nacional. Uma das formas de controle que a sua coordenação nacional possui sobre a correção, levada a cabo pelos professores que as aplicaram, é a do envio, por parte destes professores, das 10 melhores provas de cada um dos três níveis nos quais a Olimpíada é realizada. Assim, e dado ao caráter discursivo das provas da Olimpíada, a sua coordenação nacional dispõe de um vasto acervo sobre as concepções, certas ou erradas, dos alunos que dela participaram. Este conjunto de dados jamais fora explorado até o presente trabalho que pretende ser, neste sentido, inaugural. Nas provas dos níveis I e II da V OBA, que abrangem todo o conjunto do ensino fundamental, uma das questões versava sobre a noção que os alunos tinham sobre como os habitantes da Terra situam-se sobre sua superfície. Fizemos um estudo sistemático das respostas a esta questão. Primeiramente, todas as modalidades de erros foram levantadas; em seguida, realizamos uma distribuição por idade tanto do índice de acerto como da ocorrência e da distribuição de cada erro. Como resultado, obtivemos que, em ambos os níveis: 1) as concepções errôneas não se apresentam distribuídas aleatoriamente; 2) há uma concentração no tipo de erro apresentado; 3) há uma tendência de progressivo acerto com relação à idade quando os dados são considerados em termos nacionais; 4) esta tendência nacional é fraca ou mesmo não é verificada em alguns dos estados com maior patamar de acerto.

  4. Extraction of high-quality bacterial RNA from infected leaf tissue for bacterial in planta gene expression analysis by multiplexed fluorescent Northern hybridization.

    PubMed

    Schenk, Alexander; Weingart, Helge; Ullrich, Matthias S

    2008-03-01

    Plant pathogenic bacteria possess a large number of genes that allow them to grow and cause disease on plants. In planta gene expression analysis is important to understand the impact of these genes on bacterial virulence. A new mRNA-based approach using multiplexed Northern hybridization was developed. High-quality bacterial and plant total RNA was successfully isolated from leaf tissue infiltrated with Pseudomonas syringae. The procedure employs a new extraction buffer formulation containing glycine, sodium dodecylsulphate, cetyltrimethylammonium bromide, high-molecular-weight polyethylene glycol and beta-mercaptoethanol. Cell lysis and classical acid-phenol extraction steps followed by LiCl precipitation yielded large amounts of total RNA of high purity and integrity. Multiplexing of DIG and chemically fluorescently labelled RNA probes was developed and expression data were normalized using the 23S rRNA gene as reference. The method was validated by studying in planta expression of the P. syringae genes mucD, cmaA, cfl, corR, corS and corP comprising a selection of highly expressed biosynthetic and low-expressed regulatory genes. The method was assessed regarding its sensitivity and might by useful for studying a variety of plant-microbe interactions.

  5. A proline iminopeptidase gene upregulated in planta by a LuxR homologue is essential for pathogenicity of Xanthomonas campestris pv. campestris.

    PubMed

    Zhang, Lili; Jia, Yantao; Wang, Li; Fang, Rongxiang

    2007-07-01

    Expression of bacterial genes is often regulated by complex mechanisms, some of which involve host cues. Analysis of the Xanthomonas campestris pv. campestris (Xcc) genome sequence revealed the presence of an xccR/pip locus. The upstream gene xccR is a luxR homologue, while pip codes for a proline iminopeptidase. A lux box-like element, named luxXc box, locates in the pip promoter region. In this work, we show that disruption of either xccR or pip resulted in significantly attenuated virulence of Xcc. Under medium culture conditions, the pip expression was significantly enhanced by overexpression of XccR and the luxXc box is necessary for this enhancement. We further show that expression of a pip promoter-gusA fusion either inserted in the bacterial chromosome or resided in a plasmid was markedly induced when the bacteria grew in planta. Disruption of either xccR or the luxXc box abolished the in planta induction, while disruption of pip enhanced the induction. Taken together, these data demonstrate that pip is indispensable for Xcc virulence and suggest a model for Xcc-host interaction in which the pathogen senses some host factor(s) to activate XccR that subsequently interacts with the luxXc box to induce the expression of pip for facilitating Xcc infection.

  6. Mobilization of horizontally acquired island 2 is induced in planta in the phytopathogen Pectobacterium atrosepticum SCRI1043 and involves the putative relaxase ECA0613 and quorum sensing.

    PubMed

    Vanga, Bhanupratap R; Ramakrishnan, Pavithra; Butler, Ruth C; Toth, Ian K; Ronson, Clive W; Jacobs, Jeanne M E; Pitman, Andrew R

    2015-11-01

    Integrative and conjugative elements (ICEs) contribute to the rapid evolution of bacterial pathogens via horizontal gene transfer of virulence determinants. ICEs have common mechanisms for transmission, yet the cues triggering this process under natural environmental or physiological conditions are largely unknown. In this study, mobilization of the putative ICE horizontally acquired island 2 (HAI2), present in the chromosome of the phytopathogen Pectobacterium atrosepticum SCRI1043, was examined during infection of the host plant potato. Under these conditions, mobilization of HAI2 increased markedly compared with in vitro cultures. In planta-induced mobilization of HAI2 was regulated by quorum sensing and involved the putative ICE-encoded relaxase ECA0613. Disruption of ECA0613 also reduced transcription of genes involved in production of coronafacic acid (Cfa), the major virulence factor harboured on HAI2, whereas their expression was unaffected in the quorum-sensing (expI) mutant. Thus, suppression of cfa gene expression was not regulated by the mobilization of the ICE per se, but was due directly to inactivation of the relaxase. The identification of genetic factors associated solely with in planta mobilization of an ICE demonstrates that this process is highly adapted to the natural environment of the bacterial host and can influence the expression of virulence determinants.

  7. The major histocompatibility complex genes impact pain response in DA and DA.1U rats.

    PubMed

    Guo, Yuan; Yao, Fan-Rong; Cao, Dong-Yuan; Li, Li; Wang, Hui-Sheng; Xie, Wen; Zhao, Yan

    2015-08-01

    Our recent studies have shown that the difference in basal pain sensitivity to mechanical and thermal stimulation between Dark-Agouti (DA) rats and a novel congenic DA.1U rats is major histocompatibility complex (MHC) genes dependent. In the present study, we further used DA and DA.1U rats to investigate the role of MHC genes in formalin-induced pain model by behavioral, electrophysiological and immunohistochemical methods. Behavioral results showed biphasic nociceptive behaviors increased significantly following the intraplantar injection of formalin in the hindpaw of DA and DA.1U rats. The main nociceptive behaviors were lifting and licking, especially in DA rats (P<0.001 and P<0.01). The composite pain scores (CPS) in DA rats were significantly higher than those in DA.1U rats in both phases of the formalin test (P<0.01). Electrophysiological results also showed the biphasic increase in discharge rates of C and Aδ fibers of L5 dorsal root in the two strains, and the net change of the discharge rate of DA rats was significantly higher than that of DA.1U rats (P<0.05). The mechanical thresholds decreased after formalin injection in both strains (P<0.01), and the net change in the mechanical threshold in DA was greater than that in DA.1U rats (P<0.05). The expression of RT1-B, representation of MHC class II molecule, in laminae I-II of L4/5 spinal cord in DA rats was significantly higher than that in DA.1U rats in the respective experimental group (P<0.05). These results suggested that both DA and DA.1U rats exhibited nociceptive responses in formalin-induced pain model and DA rats were more sensitive to noxious chemical stimulus than DA.1U rats, indicating that MHC genes might contribute to the difference in pain sensitivity.

  8. Purification of Two Novel Sugar Acid-binding Lectins from Haplomitrium Mnioides (bryophyte, Plantae) and their Preliminary Characterization.

    PubMed

    Masuzaki, Hiroaki; Hosono, Masahiro; Nitta, Kazuo

    2017-01-01

    Two novel sugar acid-binding lectins were purified from Haplomitrium mnioides (Lindb.) Schust. using a procedure consisting of ammonium sulfate precipitation, G-50 gel filtration, hydroxyapatite chromatography, and HW-50 gel filtration. We reported their partial physicochemical properties: molecular weight, affinity for carbohydrates and organic acids, pH stability, and dependence of their hemagglutination activity on metal ions. We also determined their N-terminal amino acid sequences. H. mnioides lectins (HMLs) were monomers (one with a molecular weight of approximately 27 kDa, and the other with a molecular weight of approximately 105 kDa) under both nonreducing and reducing conditions. They were named HML27 and HML105, respectively. Both HMLs had an affinity for N-acetylneuraminic acid, D-glucuronic acid, D-glucaric acid, bovine submaxillary mucin, heparin, and organic acids, such as citrate, 2-oxoglutaric acid, and D-2-hydroxyglutarate. Furthermore, HML27 had an affinity for α-D-galacturonic acid, D-malate, L-malate, and pyruvate, while HML105 had an affinity for D-gluconic acid. HML27 and HML105 are novel plant lectins: they have an affinity for sugar acids and organic acids and specifically recognize the carboxyl group, and there is no homology between their N-terminal amino acid sequences and those of the previously described lectins and agglutinins.

  9. RNA-Seq Revealed Differences in Transcriptomes between 3ADON and 15ADON Populations of Fusarium graminearum In Vitro and In Planta

    PubMed Central

    Puri, Krishna D.; Yan, Changhui; Leng, Yueqiang

    2016-01-01

    Fusarium graminearum is the major causal agent of Fusarium head blight (FHB) in barley and wheat in North America. The fungus not only causes yield loss of the crops but also produces harmful trichothecene mycotoxins [Deoxynivalenol (DON) and its derivatives-3-acetyldeoxynivalenol (3ADON) and 15-acetyldeoxynivalenol (15ADON), and nivalenol (NIV)] that contaminate grains. Previous studies showed a dramatic increase of 3ADON-producing isolates with higher aggressiveness and DON production than the 15ADON-producing isolates in North America. However, the genetic and molecular basis of differences between the two types of isolates is unclear. In this study, we compared transcriptomes of the 3ADON and 15ADON isolates in vitro (in culture media) and in planta (during infection on the susceptible wheat cultivar ‘Briggs’) using RNA-sequencing. The in vitro gene expression comparison identified 479 up-regulated and 801 down-regulated genes in the 3ADON isolates; the up-regulated genes were mainly involved in C-compound and carbohydrate metabolism (18.6%), polysaccharide metabolism (7.7%) or were of unknown functions (57.6%). The in planta gene expression analysis revealed that 185, 89, and 62 genes were up-regulated in the 3ADON population at 48, 96, and 144 hours after inoculation (HAI), respectively. The up-regulated genes were significantly enriched in functions for cellular import, C-compound and carbohydrate metabolism, allantoin and allantoate transport at 48 HAI, for detoxification and virulence at 96 HAI, and for metabolism of acetic acid derivatives, detoxification, and cellular import at 144 HAI. Comparative analyses of in planta versus in vitro gene expression further revealed 2,159, 1,981 and 2,095 genes up-regulated in the 3ADON isolates, and 2,415, 2,059 and 1,777 genes up-regulated in the 15ADON isolates at the three time points after inoculation. Collectively, our data provides a foundation for further understanding of molecular mechanisms involved in

  10. RNA-Seq Revealed Differences in Transcriptomes between 3ADON and 15ADON Populations of Fusarium graminearum In Vitro and In Planta.

    PubMed

    Puri, Krishna D; Yan, Changhui; Leng, Yueqiang; Zhong, Shaobin

    2016-01-01

    Fusarium graminearum is the major causal agent of Fusarium head blight (FHB) in barley and wheat in North America. The fungus not only causes yield loss of the crops but also produces harmful trichothecene mycotoxins [Deoxynivalenol (DON) and its derivatives-3-acetyldeoxynivalenol (3ADON) and 15-acetyldeoxynivalenol (15ADON), and nivalenol (NIV)] that contaminate grains. Previous studies showed a dramatic increase of 3ADON-producing isolates with higher aggressiveness and DON production than the 15ADON-producing isolates in North America. However, the genetic and molecular basis of differences between the two types of isolates is unclear. In this study, we compared transcriptomes of the 3ADON and 15ADON isolates in vitro (in culture media) and in planta (during infection on the susceptible wheat cultivar 'Briggs') using RNA-sequencing. The in vitro gene expression comparison identified 479 up-regulated and 801 down-regulated genes in the 3ADON isolates; the up-regulated genes were mainly involved in C-compound and carbohydrate metabolism (18.6%), polysaccharide metabolism (7.7%) or were of unknown functions (57.6%). The in planta gene expression analysis revealed that 185, 89, and 62 genes were up-regulated in the 3ADON population at 48, 96, and 144 hours after inoculation (HAI), respectively. The up-regulated genes were significantly enriched in functions for cellular import, C-compound and carbohydrate metabolism, allantoin and allantoate transport at 48 HAI, for detoxification and virulence at 96 HAI, and for metabolism of acetic acid derivatives, detoxification, and cellular import at 144 HAI. Comparative analyses of in planta versus in vitro gene expression further revealed 2,159, 1,981 and 2,095 genes up-regulated in the 3ADON isolates, and 2,415, 2,059 and 1,777 genes up-regulated in the 15ADON isolates at the three time points after inoculation. Collectively, our data provides a foundation for further understanding of molecular mechanisms involved in

  11. Use of attenuated total reflection Fourier transform infrared (ATR FT-IR) spectroscopy in direct, non-destructive, and rapid assessment of developmental cotton fibers grown in planta and in culture

    USDA-ARS?s Scientific Manuscript database

    Cotton fibers are routinely harvested from cotton plants (in planta), and their end-use qualities depend on their development stages. Cotton fibers are also cultured at controlled laboratory environments, so that cotton researchers can investigate many aspects of experimental protocols in cotton bre...

  12. La EPA aplica un criterio flexible en cuanto a las normas que rigen las plantas eléctricas móviles importadas para usarse en Puerto Rico y las Islas Vírgenes Estadounidenses

    EPA Pesticide Factsheets

    Comunicado de prensa de la EPA: La EPA aplica un criterio flexible en cuanto a las normas que rigen las plantas eléctricas móviles importadas para usarse en Puerto Rico y las Islas Vírgenes Estadounidenses

  13. In vitro and in planta interaction evidence between Nicotiana tabacum thaumatin-like protein 1 (TLP1) and cucumber mosaic virus proteins.

    PubMed

    Kim, Min Jung; Ham, Byung-Kook; Kim, Hwa Ran; Lee, In-Ju; Kim, Young Jin; Ryu, Ki Hyun; Park, Young In; Paek, Kyung-Hee

    2005-12-01

    Using a yeast two-hybrid system, we identified a plant cellular factor that interacts with the proteins of the Cucumber mosaic virus (CMV). Initially 14 candidate genes were isolated from Nicotiana tabacum, using a full-length CMV 1a gene as bait. Among the candidate genes, two were encoding thaumatin-like proteins (TLP), and were designated as Nicotiana tabacum thaumatin-like protein 1 (NtTLP1). Consistent with this observation, recombinant GST-NtTLP1 protein, which was expressed and purified in E. coli, bound tightly to CMV 1a in vitro. In planta interaction was also verified via co-immunoprecipitation. Additionally, NtTLP1 specifically interacted with the CMV movement-related proteins, movement protein and coat protein, in yeast. Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that the expression of NtTLP1 increased as the result of CMV inoculation.

  14. Forward genetic in planta screen for identification of plant-protective traits of Sphingomonas sp. strain Fr1 against Pseudomonas syringae DC3000.

    PubMed

    Vogel, Christine; Innerebner, Gerd; Zingg, Judith; Guder, Jan; Vorholt, Julia A

    2012-08-01

    Sphingomonas sp. strain Fr1 has recently been shown to protect Arabidopsis thaliana against the bacterial leaf pathogen Pseudomonas syringae DC3000. Here, we describe a forward genetic in planta screen to identify genes in Sphingomonas sp. Fr1 necessary for this effect. About 5,000 Sphingomonas sp. Fr1 mini-Tn5 mutants were assayed for a defect in plant protection against a luxCDABE-tagged P. syringae DC3000 derivative in a space-saving 24-well plate system. The bioluminescence of the pathogen was used as the indicator of pathogen proliferation and allowed for the identification of Sphingomonas sp. Fr1 mutants that had lost the ability to restrict pathogen growth before disease symptoms were visible. Potential candidates were validated using the same miniaturized experimental system. Of these mutants, 10 were confirmed as plant protection defective yet colonization competent. The mutants were subsequently evaluated in a previously described standard microbox system, and plants showed enhanced disease phenotypes after pathogen infection relative to those inoculated with the parental strain as a control. However, the disease severities were lower than those observed for control plants that were grown axenically prior to pathogen challenge, which suggests that several traits may contribute to plant protection. Transposon insertion sites of validated mutants with defects in plant protection were determined and mapped to 7 distinct genomic regions. In conclusion, the established screening protocol allowed us to identify mutations that affect plant protection, and it opens the possibility to uncover traits important for in planta microbe-microbe interactions.

  15. The Small Subunit of Snapdragon Geranyl Diphosphate Synthase Modifies the Chain Length Specificity of Tobacco Geranylgeranyl Diphosphate Synthase in Planta[W

    PubMed Central

    Orlova, Irina; Nagegowda, Dinesh A.; Kish, Christine M.; Gutensohn, Michael; Maeda, Hiroshi; Varbanova, Marina; Fridman, Eyal; Yamaguchi, Shinjiro; Hanada, Atsushi; Kamiya, Yuji; Krichevsky, Alexander; Citovsky, Vitaly; Pichersky, Eran; Dudareva, Natalia

    2009-01-01

    Geranyl diphosphate (GPP), the precursor of many monoterpene end products, is synthesized in plastids by a condensation of dimethylallyl diphosphate and isopentenyl diphosphate (IPP) in a reaction catalyzed by homodimeric or heterodimeric GPP synthase (GPPS). In the heterodimeric enzymes, a noncatalytic small subunit (GPPS.SSU) determines the product specificity of the catalytic large subunit, which may be either an active geranylgeranyl diphosphate synthase (GGPPS) or an inactive GGPPS-like protein. Here, we show that expression of snapdragon (Antirrhinum majus) GPPS.SSU in tobacco (Nicotiana tabacum) plants increased the total GPPS activity and monoterpene emission from leaves and flowers, indicating that the introduced catalytically inactive GPPS.SSU found endogenous large subunit partner(s) and formed an active snapdragon/tobacco GPPS in planta. Bimolecular fluorescence complementation and in vitro enzyme analysis of individual and hybrid proteins revealed that two of four GGPPS-like candidates from tobacco EST databases encode bona fide GGPPS that can interact with snapdragon GPPS.SSU and form a functional GPPS enzyme in plastids. The formation of chimeric GPPS in transgenic plants also resulted in leaf chlorosis, increased light sensitivity, and dwarfism due to decreased levels of chlorophylls, carotenoids, and gibberellins. In addition, these transgenic plants had reduced levels of sesquiterpene emission, suggesting that the export of isoprenoid intermediates from the plastids into the cytosol was decreased. These results provide genetic evidence that GPPS.SSU modifies the chain length specificity of phylogenetically distant GGPPS and can modulate IPP flux distribution between GPP and GGPP synthesis in planta. PMID:20028839

  16. Transient Expression of Secretory IgA In Planta is Optimal Using a Multi-Gene Vector and may be Further Enhanced by Improving Joining Chain Incorporation

    PubMed Central

    Westerhof, Lotte B.; Wilbers, Ruud H. P.; van Raaij, Debbie R.; van Wijk, Christina Z.; Goverse, Aska; Bakker, Jaap; Schots, Arjen

    2016-01-01

    Secretory IgA (sIgA) is a crucial antibody in host defense at mucosal surfaces. It is a promising antibody isotype in a variety of therapeutic settings such as passive vaccination and treatment of inflammatory disorders. However, heterologous production of this heteromultimeric protein complex is still suboptimal. The challenge is the coordinate expression of the four required polypeptides; the alpha heavy chain, the light chain, the joining chain, and part of the polymeric-Ig-receptor called the secretory component, in a 4:4:1:1 ratio. We evaluated the transient expression of three sIgAκ variants, harboring the heavy chain isotype α1, α2m1, or α2m2, of the clinical antibody Ustekinumab in planta. Ustekinumab is directed against the p40 subunit that is shared by the pro-inflammatory cytokines interleukin (IL)-12 and IL-23. A sIgA variant of this antibody may enable localized treatment of inflammatory bowel disease. Of the three different sIgA variants we obtained the highest yield with sIgA1κ reaching up to 373 μg sIgA/mg total soluble protein. The use of a multi-cassette vector containing all four expression cassettes was most efficient. However, not the expression strategy, but the incorporation of the joining chain turned out to be the limiting step for sIgA production. Our data demonstrate that transient expression in planta is suitable for the economic production of heteromultimeric protein complexes such as sIgA. PMID:26793201

  17. Transient Expression of Secretory IgA In Planta is Optimal Using a Multi-Gene Vector and may be Further Enhanced by Improving Joining Chain Incorporation.

    PubMed

    Westerhof, Lotte B; Wilbers, Ruud H P; van Raaij, Debbie R; van Wijk, Christina Z; Goverse, Aska; Bakker, Jaap; Schots, Arjen

    2015-01-01

    Secretory IgA (sIgA) is a crucial antibody in host defense at mucosal surfaces. It is a promising antibody isotype in a variety of therapeutic settings such as passive vaccination and treatment of inflammatory disorders. However, heterologous production of this heteromultimeric protein complex is still suboptimal. The challenge is the coordinate expression of the four required polypeptides; the alpha heavy chain, the light chain, the joining chain, and part of the polymeric-Ig-receptor called the secretory component, in a 4:4:1:1 ratio. We evaluated the transient expression of three sIgAκ variants, harboring the heavy chain isotype α1, α2m1, or α2m2, of the clinical antibody Ustekinumab in planta. Ustekinumab is directed against the p40 subunit that is shared by the pro-inflammatory cytokines interleukin (IL)-12 and IL-23. A sIgA variant of this antibody may enable localized treatment of inflammatory bowel disease. Of the three different sIgA variants we obtained the highest yield with sIgA1κ reaching up to 373 μg sIgA/mg total soluble protein. The use of a multi-cassette vector containing all four expression cassettes was most efficient. However, not the expression strategy, but the incorporation of the joining chain turned out to be the limiting step for sIgA production. Our data demonstrate that transient expression in planta is suitable for the economic production of heteromultimeric protein complexes such as sIgA.

  18. Forward Genetic In Planta Screen for Identification of Plant-Protective Traits of Sphingomonas sp. Strain Fr1 against Pseudomonas syringae DC3000

    PubMed Central

    Vogel, Christine; Innerebner, Gerd; Zingg, Judith; Guder, Jan

    2012-01-01

    Sphingomonas sp. strain Fr1 has recently been shown to protect Arabidopsis thaliana against the bacterial leaf pathogen Pseudomonas syringae DC3000. Here, we describe a forward genetic in planta screen to identify genes in Sphingomonas sp. Fr1 necessary for this effect. About 5,000 Sphingomonas sp. Fr1 mini-Tn5 mutants were assayed for a defect in plant protection against a luxCDABE-tagged P. syringae DC3000 derivative in a space-saving 24-well plate system. The bioluminescence of the pathogen was used as the indicator of pathogen proliferation and allowed for the identification of Sphingomonas sp. Fr1 mutants that had lost the ability to restrict pathogen growth before disease symptoms were visible. Potential candidates were validated using the same miniaturized experimental system. Of these mutants, 10 were confirmed as plant protection defective yet colonization competent. The mutants were subsequently evaluated in a previously described standard microbox system, and plants showed enhanced disease phenotypes after pathogen infection relative to those inoculated with the parental strain as a control. However, the disease severities were lower than those observed for control plants that were grown axenically prior to pathogen challenge, which suggests that several traits may contribute to plant protection. Transposon insertion sites of validated mutants with defects in plant protection were determined and mapped to 7 distinct genomic regions. In conclusion, the established screening protocol allowed us to identify mutations that affect plant protection, and it opens the possibility to uncover traits important for in planta microbe-microbe interactions. PMID:22660707

  19. Autocorrelation descriptor improvements for QSAR: 2DA_Sign and 3DA_Sign

    NASA Astrophysics Data System (ADS)

    Sliwoski, Gregory; Mendenhall, Jeffrey; Meiler, Jens

    2016-03-01

    Quantitative structure-activity relationship (QSAR) is a branch of computer aided drug discovery that relates chemical structures to biological activity. Two well established and related QSAR descriptors are two- and three-dimensional autocorrelation (2DA and 3DA). These descriptors encode the relative position of atoms or atom properties by calculating the separation between atom pairs in terms of number of bonds (2DA) or Euclidean distance (3DA). The sums of all values computed for a given small molecule are collected in a histogram. Atom properties can be added with a coefficient that is the product of atom properties for each pair. This procedure can lead to information loss when signed atom properties are considered such as partial charge. For example, the product of two positive charges is indistinguishable from the product of two equivalent negative charges. In this paper, we present variations of 2DA and 3DA called 2DA_Sign and 3DA_Sign that avoid information loss by splitting unique sign pairs into individual histograms. We evaluate these variations with models trained on nine datasets spanning a range of drug target classes. Both 2DA_Sign and 3DA_Sign significantly increase model performance across all datasets when compared with traditional 2DA and 3DA. Lastly, we find that limiting 3DA_Sign to maximum atom pair distances of 6 Å instead of 12 Å further increases model performance, suggesting that conformational flexibility may hinder performance with longer 3DA descriptors. Consistent with this finding, limiting the number of bonds in 2DA_Sign from 11 to 5 fails to improve performance.

  20. Autocorrelation descriptor improvements for QSAR: 2DA_Sign and 3DA_Sign.

    PubMed

    Sliwoski, Gregory; Mendenhall, Jeffrey; Meiler, Jens

    2016-03-01

    Quantitative structure-activity relationship (QSAR) is a branch of computer aided drug discovery that relates chemical structures to biological activity. Two well established and related QSAR descriptors are two- and three-dimensional autocorrelation (2DA and 3DA). These descriptors encode the relative position of atoms or atom properties by calculating the separation between atom pairs in terms of number of bonds (2DA) or Euclidean distance (3DA). The sums of all values computed for a given small molecule are collected in a histogram. Atom properties can be added with a coefficient that is the product of atom properties for each pair. This procedure can lead to information loss when signed atom properties are considered such as partial charge. For example, the product of two positive charges is indistinguishable from the product of two equivalent negative charges. In this paper, we present variations of 2DA and 3DA called 2DA_Sign and 3DA_Sign that avoid information loss by splitting unique sign pairs into individual histograms. We evaluate these variations with models trained on nine datasets spanning a range of drug target classes. Both 2DA_Sign and 3DA_Sign significantly increase model performance across all datasets when compared with traditional 2DA and 3DA. Lastly, we find that limiting 3DA_Sign to maximum atom pair distances of 6 Å instead of 12 Å further increases model performance, suggesting that conformational flexibility may hinder performance with longer 3DA descriptors. Consistent with this finding, limiting the number of bonds in 2DA_Sign from 11 to 5 fails to improve performance.

  1. BacS: an abundant bacteroid protein in Rhizobium etli whose expression ex planta requires nifA.

    PubMed

    Jahn, Olivia J; Davila, Guillermo; Romero, David; Noel, K Dale

    2003-01-01

    Rhizobium etli CFN42 bacteroids from bean nodules possessed an abundant 16-kDa protein (BacS) that was found in the membrane pellet after cell disruption. This protein was not detected in bacteria cultured in tryptone-yeast extract. In minimal media, it was produced at low oxygen concentration but not in a mutant whose nifA was disrupted. N-terminal sequencing of the protein led to isolation of a bacS DNA fragment. DNA hybridization and nucleotide sequencing revealed three copies of the bacS gene, all residing on the main symbiotic plasmid of strain CFN42. A stretch of 304 nucleotides, exactly conserved upstream of all three bacS open reading frames, had very close matches with the NifA and sigma 54 consensus binding sequences. The only bacS homology in the genetic sequence databases was to three hypothetical proteins of unknown function, all from rhizobial species. Mutation and genetic complementation indicated that each of the bacS genes gives rise to a BacS polypeptide. Mutants disrupted or deleted in all three genes did not produce the BacS polypeptide but were Nod+ and Fix+ on Phaseolus vulgaris.

  2. Efeitos do binarismo não resolvido na determinação da função de massa de aglomerados

    NASA Astrophysics Data System (ADS)

    Kerber, L. O.; Santiago, B. X.

    2003-08-01

    Através de simulações numéricas buscamos quantificar os efeitos que o binarismo não resolvido causa na determinação da função de massa (MF) de aglomerados estelares. Geramos diagramas cor-magnitude (CMDs) artificiais simulando uma população única, caracterizada por estrelas de mesma idade e composição quí mica, com uma fração de binárias não resolvidas e distribuição em massa das estrelas dada por uma MF do tipo lei de potência. A presença de pares de estrelas não resolvidos faz com que a MF obtida da função de luminosidade (LF) tenha a têndencia de ser mais plana do que a MF que gerou o CMD artificial. Propomos um tratamento de correção para tal efeito. Outro efeito relacionado diz respeito ao alargamento do CMD, que apresenta-se como um indicador do número total de estrelas no domí nio de baixas massas (m < 0.6M¤). Todos os resultados acima possuem uma forte dependência com os erros fotométricos e estão baseados na hipótese de que ambas estrelas do par não resolvido são sorteadas de uma mesma MF de forma independente. O objetivo final é aplicarmos o tratamento aqui desenvolvido para implementarmos a análise da nossa amostra de aglomerados ricos da Grande Nuvem de Magalhães.

  3. The potentially dangerous asteroid 2012 DA14

    NASA Astrophysics Data System (ADS)

    Wlodarczyk, I.

    2012-12-01

    We present computing methods that allow us to study the behaviour of the dynamically interesting potentially dangerous asteroid 2012 DA14. Using the freely available ORBFIT software, we can follow the orbit of the asteroid backward and forward in the future, searching for close approaches to the Earth that might lead to possible impacts. The possible impact orbit for 2026 is computed. We show that it should be possible to recover asteroid 2012 DA14, mainly in 2013 February. It is highly unlikely that asteroid 2012 DA14 will hit any geosynchronous satellites during its close approach on 2013 February 15.

  4. Origin of the DA and non-DA white dwarf stars

    NASA Technical Reports Server (NTRS)

    Shipman, Harry L.

    1989-01-01

    Various proposals for the bifurcation of the white dwarf cooling sequence are reviewed. 'Primordial' theories, in which the basic bifurcation of the white dwarf sequence is rooted in events predating the white dwarf stage of stellar evolution, are discussed, along with the competing 'mixing' theories in which processes occurring during the white dwarf stage are responsible for the existence of DA or non-DA stars. A new proposal is suggested, representing a two-channel scenario. In the DA channel, some process reduces the hydrogen layer mass to the value of less than 10 to the -7th. The non-DA channel is similar to that in the primordial scenario. These considerations suggest that some mechanism operates in both channels to reduce the thickness of the outermost layer of the white dwarf. It is also noted that accretion from the interstellar medium has little to do with whether a particular white dwarf becomes a DA or a non-DA star.

  5. An Extension of PPLS-DA for Classification and Comparison to Ordinary PLS-DA

    PubMed Central

    Telaar, Anna; Liland, Kristian Hovde; Repsilber, Dirk; Nürnberg, Gerd

    2013-01-01

    Classification studies are widely applied, e.g. in biomedical research to classify objects/patients into predefined groups. The goal is to find a classification function/rule which assigns each object/patient to a unique group with the greatest possible accuracy (classification error). Especially in gene expression experiments often a lot of variables (genes) are measured for only few objects/patients. A suitable approach is the well-known method PLS-DA, which searches for a transformation to a lower dimensional space. Resulting new components are linear combinations of the original variables. An advancement of PLS-DA leads to PPLS-DA, introducing a so called ‘power parameter’, which is maximized towards the correlation between the components and the group-membership. We introduce an extension of PPLS-DA for optimizing this power parameter towards the final aim, namely towards a minimal classification error. We compare this new extension with the original PPLS-DA and also with the ordinary PLS-DA using simulated and experimental datasets. For the investigated data sets with weak linear dependency between features/variables, no improvement is shown for PPLS-DA and for the extensions compared to PLS-DA. A very weak linear dependency, a low proportion of differentially expressed genes for simulated data, does not lead to an improvement of PPLS-DA over PLS-DA, but our extension shows a lower prediction error. On the contrary, for the data set with strong between-feature collinearity and a low proportion of differentially expressed genes and a large total number of genes, the prediction error of PPLS-DA and the extensions is clearly lower than for PLS-DA. Moreover we compare these prediction results with results of support vector machines with linear kernel and linear discriminant analysis. PMID:23408965

  6. An extension of PPLS-DA for classification and comparison to ordinary PLS-DA.

    PubMed

    Telaar, Anna; Liland, Kristian Hovde; Repsilber, Dirk; Nürnberg, Gerd

    2013-01-01

    Classification studies are widely applied, e.g. in biomedical research to classify objects/patients into predefined groups. The goal is to find a classification function/rule which assigns each object/patient to a unique group with the greatest possible accuracy (classification error). Especially in gene expression experiments often a lot of variables (genes) are measured for only few objects/patients. A suitable approach is the well-known method PLS-DA, which searches for a transformation to a lower dimensional space. Resulting new components are linear combinations of the original variables. An advancement of PLS-DA leads to PPLS-DA, introducing a so called 'power parameter', which is maximized towards the correlation between the components and the group-membership. We introduce an extension of PPLS-DA for optimizing this power parameter towards the final aim, namely towards a minimal classification error. We compare this new extension with the original PPLS-DA and also with the ordinary PLS-DA using simulated and experimental datasets. For the investigated data sets with weak linear dependency between features/variables, no improvement is shown for PPLS-DA and for the extensions compared to PLS-DA. A very weak linear dependency, a low proportion of differentially expressed genes for simulated data, does not lead to an improvement of PPLS-DA over PLS-DA, but our extension shows a lower prediction error. On the contrary, for the data set with strong between-feature collinearity and a low proportion of differentially expressed genes and a large total number of genes, the prediction error of PPLS-DA and the extensions is clearly lower than for PLS-DA. Moreover we compare these prediction results with results of support vector machines with linear kernel and linear discriminant analysis.

  7. A plastid-targeted heat shock cognate 70 kDa protein interacts with the Abutilon mosaic virus movement protein

    SciTech Connect

    Krenz, Bjoern; Windeisen, Volker; Wege, Christina; Jeske, Holger; Kleinow, Tatjana

    2010-05-25

    The movement protein (MP) of bipartite geminiviruses facilitates cell-to-cell as well as long-distance transport within plants and influences viral pathogenicity. Yeast two-hybrid assays identified a chaperone, the nuclear-encoded and plastid-targeted heat shock cognate 70 kDa protein (cpHSC70-1) of Arabidopsis thaliana, as a potential binding partner for the Abutilon mosaic virus (AbMV) MP. In planta, bimolecular fluorescence complementation (BiFC) analysis showed cpHSC70-1/MP complexes and MP homooligomers at the cell periphery and co-localized with chloroplasts. BiFC revealed cpHSC70-1 oligomers associated with chloroplasts, but also distributed at the cellular margin and in filaments arising from plastids reminiscent of stromules. Silencing the cpHSC70 gene of Nicotiana benthamiana using an AbMV DNA A-derived gene silencing vector induced minute white leaf areas, which indicate an effect on chloroplast stability. Although AbMV DNA accumulated within chlorotic spots, a spatial restriction of these occurred, suggesting a functional relevance of the MP-chaperone interaction for viral transport and symptom induction.

  8. Specific in planta recognition of two GKLR proteins of the downy mildew Bremia lactucae revealed in a large effector screen in lettuce.

    PubMed

    Stassen, Joost H M; den Boer, Erik; Vergeer, Pim W J; Andel, Annemiek; Ellendorff, Ursula; Pelgrom, Koen; Pel, Mathieu; Schut, Johan; Zonneveld, Olaf; Jeuken, Marieke J W; Van den Ackerveken, Guido

    2013-11-01

    Breeding lettuce (Lactuca sativa) for resistance to the downy mildew pathogen Bremia lactucae is mainly achieved by introgression of dominant downy mildew resistance (Dm) genes. New Bremia races quickly render Dm genes ineffective, possibly by mutation of recognized host-translocated effectors or by suppression of effector-triggered immunity. We have previously identified 34 potential RXLR(-like) effector proteins of B. lactucae that were here tested for specific recognition within a collection of 129 B. lactucae-resistant Lactuca lines. Two effectors triggered a hypersensitive response: BLG01 in 52 lines, predominantly L. saligna, and BLG03 in two L. sativa lines containing Dm2 resistance. The N-terminal sequences of BLG01 and BLG03, containing the signal peptide and GKLR variant of the RXLR translocation motif, are not required for in planta recognition but function in effector delivery. The locus responsible for BLG01 recognition maps to the bottom of lettuce chromosome 9, whereas recognition of BLG03 maps in the RGC2 cluster on chromosome 2. Lactuca lines that recognize the BLG effectors are not resistant to Bremia isolate Bl:24 that expresses both BLG genes, suggesting that Bl:24 can suppress the triggered immune responses. In contrast, lettuce segregants displaying Dm2-mediated resistance to Bremia isolate Bl:5 are responsive to BLG03, suggesting that BLG03 is a candidate Avr2 protein.

  9. Cell wall targeted in planta iron accumulation enhances biomass conversion and seed iron concentration in Arabidopsis and rice

    SciTech Connect

    Yang, Haibing; Wei, Hui; Ma, Guojie; Antunes, Mauricio S.; Vogt, Stefan; Cox, Joseph; Zhang, Xiao; Liu, Xiping; Bu, Lintao; Gleber, S. Charlotte; Carpita, Nicholas C.; Makowski, Lee; Himmel, Michael E.; Tucker, Melvin P.; McCann, Maureen C.; Murphy, Angus S.; Peer, Wendy A.

    2016-04-07

    Conversion of nongrain biomass into liquid fuel is a sustainable approach to energy demands as global population increases. Previously, we showed that iron can act as a catalyst to enhance the degradation of lignocellulosic biomass for biofuel production. However, direct addition of iron catalysts to biomass pretreatment is diffusion-limited, would increase the cost and complexity of biorefinery unit operations and may have deleterious environmental impacts. Here, we show a new strategy for in planta accumulation of iron throughout the volume of the cell wall where iron acts as a catalyst in the deconstruction of lignocellulosic biomass. We engineered CBM-IBP fusion polypeptides composed of a carbohydrate-binding module family 11 (CBM11) and an iron-binding peptide (IBP) for secretion into Arabidopsis and rice cell walls. CBM-IBP transformed Arabidopsis and rice plants show significant increases in iron accumulation and biomass conversion compared to respective controls. Further, CBM-IBP rice shows a 35% increase in seed iron concentration and a 40% increase in seed yield in greenhouse experiments. CBM-IBP rice potentially could be used to address iron deficiency, the most common and widespread nutritional disorder according to the World Health Organization.

  10. The predominant circular form of avocado sunblotch viroid accumulates in planta as a free RNA adopting a rod-shaped secondary structure unprotected by tightly bound host proteins.

    PubMed

    López-Carrasco, Amparo; Flores, Ricardo

    2017-07-01

    Avocado sunblotch viroid (ASBVd), the type member of the family Avsunviroidae, replicates and accumulates in chloroplasts. Whether this minimal non-protein-coding circular RNA of 246-250 nt exists in vivo as a free nucleic acid or closely associated with host proteins remains unknown. To tackle this issue, the secondary structures of the monomeric circular (mc) (+) and (-) strands of ASBVd have been examined in silico by searching those of minimal free energy, and in vitro at single-nucleotide resolution by selective 2'-hydroxyl acylation analysed by primer extension (SHAPE). Both approaches resulted in predominant rod-like secondary structures without tertiary interactions, with the mc (+) RNA being more compact than its (-) counterpart as revealed by non-denaturing polyacryamide gel electrophoresis. Moreover, in vivo SHAPE showed that the mc ASBVd (+) form accumulates in avocado leaves as a free RNA adopting a similar rod-shaped conformation unprotected by tightly bound host proteins. Hence, the mc ASBVd (+) RNA behaves in planta like the previously studied mc (+) RNA of potato spindle tuber viroid, the type member of nuclear viroids (family Pospiviroidae), indicating that two different viroids replicating and accumulating in distinct subcellular compartments, have converged into a common structural solution. Circularity and compact secondary structures confer to these RNAs, and probably to all viroids, the intrinsic stability needed to survive in their natural habitats. However, in vivo SHAPE has not revealed the (possibly transient or loose) interactions of the mc ASBVd (+) RNA with two host proteins observed previously by UV irradiation of infected avocado leaves.

  11. Mutations in the capsid protein of Brome mosaic virus affecting encapsidation eliminate vesicle induction in planta: implications for virus cell-to-cell spread.

    PubMed

    Bamunusinghe, Devinka; Chaturvedi, Sonali; Seo, Jang-Kyun; Rao, A L N

    2013-08-01

    Positive-strand RNA viruses are known to rearrange the endomembrane network to make it more conducive for replication, maturation, or egress. Our previous transmission electron microscopic (TEM) analysis showed that ectopic expression of wild-type (wt) capsid protein (CP) of Brome mosaic virus (BMV) has an intrinsic property of modifying the endoplasmic reticulum (ER) to induce vesicles similar to those present in wt BMV infection. In this study, we evaluated the functional significance of CP-mediated vesicle induction to the BMV infection cycle in planta. Consequently, the cytopathologic changes induced by wt CP or its mutants defective in virion assembly due to mutations engineered in either N- or C-proximal domains were comparatively analyzed by TEM in two susceptible (Nicotiana benthamiana and Chenopodium quinoa) and one nonhost (N. clevelandii) plant species. The results showed that in susceptible hosts, CP-mediated ER-derived vesicle induction is contingent on the expression of encapsidation-competent CP. In contrast, unlike in N. benthamiana and C. quinoa, transient expression of wt CP in nonhost N. clevelandii plants eliminated vesicle induction. Additionally, comparative source-to-sink analysis of virus spread in leaves of N. benthamiana and N. clevelandii coexpressing wt BMV and Cucumber mosaic virus (CMV) showed that despite trans-encapsidation, CMV failed to complement the defective cell-to-cell movement of BMV. The significance and relation of CP-mediated vesicle induction to virus cell-to-cell movement are discussed.

  12. Mutations in the Capsid Protein of Brome Mosaic Virus Affecting Encapsidation Eliminate Vesicle Induction In Planta: Implications for Virus Cell-to-Cell Spread

    PubMed Central

    Bamunusinghe, Devinka; Chaturvedi, Sonali; Seo, Jang-Kyun

    2013-01-01

    Positive-strand RNA viruses are known to rearrange the endomembrane network to make it more conducive for replication, maturation, or egress. Our previous transmission electron microscopic (TEM) analysis showed that ectopic expression of wild-type (wt) capsid protein (CP) of Brome mosaic virus (BMV) has an intrinsic property of modifying the endoplasmic reticulum (ER) to induce vesicles similar to those present in wt BMV infection. In this study, we evaluated the functional significance of CP-mediated vesicle induction to the BMV infection cycle in planta. Consequently, the cytopathologic changes induced by wt CP or its mutants defective in virion assembly due to mutations engineered in either N- or C-proximal domains were comparatively analyzed by TEM in two susceptible (Nicotiana benthamiana and Chenopodium quinoa) and one nonhost (N. clevelandii) plant species. The results showed that in susceptible hosts, CP-mediated ER-derived vesicle induction is contingent on the expression of encapsidation-competent CP. In contrast, unlike in N. benthamiana and C. quinoa, transient expression of wt CP in nonhost N. clevelandii plants eliminated vesicle induction. Additionally, comparative source-to-sink analysis of virus spread in leaves of N. benthamiana and N. clevelandii coexpressing wt BMV and Cucumber mosaic virus (CMV) showed that despite trans-encapsidation, CMV failed to complement the defective cell-to-cell movement of BMV. The significance and relation of CP-mediated vesicle induction to virus cell-to-cell movement are discussed. PMID:23741003

  13. Arabidopsis DAYU/ABERRANT PEROXISOME MORPHOLOGY9 Is a Key Regulator of Peroxisome Biogenesis and Plays Critical Roles during Pollen Maturation and Germination in Planta[W

    PubMed Central

    Li, Xin-Ran; Li, Hong-Ju; Yuan, Li; Liu, Man; Shi, Dong-Qiao; Liu, Jie; Yang, Wei-Cai

    2014-01-01

    Pollen undergo a maturation process to sustain pollen viability and prepare them for germination. Molecular mechanisms controlling these processes remain largely unknown. Here, we report an Arabidopsis thaliana mutant, dayu (dau), which impairs pollen maturation and in vivo germination. Molecular analysis indicated that DAU encodes the peroxisomal membrane protein ABERRANT PEROXISOME MORPHOLOGY9 (APEM9). DAU is transiently expressed from bicellular pollen to mature pollen during male gametogenesis. DAU interacts with peroxisomal membrane proteins PEROXIN13 (PEX13) and PEX16 in planta. Consistently, both peroxisome biogenesis and peroxisome protein import are impaired in dau pollen. In addition, the jasmonic acid (JA) level is significantly decreased in dau pollen, and the dau mutant phenotype is partially rescued by exogenous application of JA, indicating that the male sterility is mainly due to JA deficiency. In addition, the phenotypic survey of peroxin mutants indicates that the PEXs most likely play different roles in pollen germination. Taken together, these data indicate that DAU/APEM9 plays critical roles in peroxisome biogenesis and function, which is essential for JA production and pollen maturation and germination. PMID:24510720

  14. In Planta Biocontrol of Pectobacterium atrosepticum by Rhodococcus erythropolis Involves Silencing of Pathogen Communication by the Rhodococcal Gamma-Lactone Catabolic Pathway.

    PubMed

    Barbey, Corinne; Crépin, Alexandre; Bergeau, Dorian; Ouchiha, Asma; Mijouin, Lily; Taupin, Laure; Orange, Nicole; Feuilloley, Marc; Dufour, Alain; Burini, Jean-François; Latour, Xavier

    2013-01-01

    The virulence of numerous Gram-negative bacteria is under the control of a quorum sensing process based on synthesis and perception of N-acyl homoserine lactones. Rhodococcus erythropolis, a Gram-positive bacterium, has recently been proposed as a biocontrol agent for plant protection against soft-rot bacteria, including Pectobacterium. Here, we show that the γ-lactone catabolic pathway of R. erythropolis disrupts Pectobacterium communication and prevents plant soft-rot. We report the first characterization and demonstration of N-acyl homoserine lactone quenching in planta. In particular, we describe the transcription of the R. erythropolis lactonase gene, encoding the key enzyme of this pathway, and the subsequent lactone breakdown. The role of this catabolic pathway in biocontrol activity was confirmed by deletion of the lactonase gene from R. erythropolis and also its heterologous expression in Escherichia coli. The γ-lactone catabolic pathway is induced by pathogen communication rather than by pathogen invasion. This is thus a novel and unusual biocontrol pathway, differing from those previously described as protecting plants from phytopathogens. These findings also suggest the existence of an additional pathway contributing to plant protection.

  15. Genetic modification of potato against microbial diseases: in vitro and in planta activity of a dermaseptin B1 derivative, MsrA2.

    PubMed

    Osusky, Milan; Osuska, Lubica; Kay, William; Misra, Santosh

    2005-08-01

    Dermaseptin B1 is a potent cationic antimicrobial peptide found in skin secretions of the arboreal frog Phyllomedusa bicolor. A synthetic derivative of dermaseptin B1, MsrA2 (N-Met-dermaseptin B1), elicited strong antimicrobial activities against various phytopathogenic fungi and bacteria in vitro. To assess its potential for plant protection, MsrA2 was expressed at low levels (1-5 microg/g of fresh tissue) in the transgenic potato (Solanum tuberosum L.) cv. Desiree. Stringent challenges of these transgenic potato plants with a variety of highly virulent fungal phytopathogens--Alternaria, Cercospora, Fusarium, Phytophthora, Pythium, Rhizoctonia and Verticillium species--and with the bacterial pathogen Erwinia carotovora demonstrated that the plants had an unusually broad-spectrum and powerful resistance to infection. MsrA2 profoundly protected both plants and tubers from diseases such as late blight, dry rot and pink rot and markedly extended the storage life of tubers. Due to these properties in planta, MsrA2 is proposed as an ideal antimicrobial peptide candidate to significantly increase resistance to phytopathogens and improve quality in a variety of crops worldwide with the potential to obviate fungicides and facilitate storage under difficult conditions.

  16. In Planta Stage-Specific Fungal Gene Profiling Elucidates the Molecular Strategies of Fusarium graminearum Growing inside Wheat Coleoptiles[W][OA

    PubMed Central

    Zhang, Xiao-Wei; Jia, Lei-Jie; Zhang, Yan; Jiang, Gang; Li, Xuan; Zhang, Dong; Tang, Wei-Hua

    2012-01-01

    The ascomycete Fusarium graminearum is a destructive fungal pathogen of wheat (Triticum aestivum). To better understand how this pathogen proliferates within the host plant, we tracked pathogen growth inside wheat coleoptiles and then examined pathogen gene expression inside wheat coleoptiles at 16, 40, and 64 h after inoculation (HAI) using laser capture microdissection and microarray analysis. We identified 344 genes that were preferentially expressed during invasive growth in planta. Gene expression profiles for 134 putative plant cell wall–degrading enzyme genes suggest that there was limited cell wall degradation at 16 HAI and extensive degradation at 64 HAI. Expression profiles for genes encoding reactive oxygen species (ROS)–related enzymes suggest that F. graminearum primarily scavenges extracellular ROS before a later burst of extracellular ROS is produced by F. graminearum enzymes. Expression patterns of genes involved in primary metabolic pathways suggest that F. graminearum relies on the glyoxylate cycle at an early stage of plant infection. A secondary metabolite biosynthesis gene cluster was specifically induced at 64 HAI and was required for virulence. Our results indicate that F. graminearum initiates infection of coleoptiles using covert penetration strategies and switches to overt cellular destruction of tissues at an advanced stage of infection. PMID:23266949

  17. Colonization process of olive tissues by Verticillium dahliae and its in planta interaction with the biocontrol root endophyte Pseudomonas fluorescens PICF7

    PubMed Central

    Prieto, Pilar; Navarro‐Raya, Carmen; Valverde‐Corredor, Antonio; Amyotte, Stefan G.; Dobinson, Katherine F.; Mercado‐Blanco, Jesús

    2009-01-01

    Summary The colonization process of Olea europaea by the defoliating pathotype of Verticillium dahliae, and the in planta interaction with the endophytic, biocontrol strain Pseudomonas fluorescens PICF7 were determined. Differential fluorescent protein tagging was used for the simultaneous visualization of P. fluorescens PICF7 and V. dahliae in olive tissues. Olive plants were bacterized with PICF7 and then transferred to V. dahliae‐infested soil. Monitoring olive colonization events by V. dahliae and its interaction with PICF7 was conducted using a non‐gnotobiotic system, confocal laser scanner microscopy and tissue vibratoming sections. A yellow fluorescently tagged V. dahliae derivative (VDAT‐36I) was obtained by Agrobacterium tumefaciens‐mediated transformation. Isolate VDAT‐36I quickly colonized olive root surface, successfully invaded root cortex and vascular tissues via macro‐ and micro‐breakages, and progressed to the aerial parts of the plant through xylem vessel cells. Strain PICF7 used root hairs as preferred penetration site, and once established on/in root tissues, hindered pathogen colonization. For the first time using this approach, the entire colonization process of a woody plant by V. dahliae is reported. Early and localized root surface and root endophytic colonization by P. fluorescens PICF7 is needed to impair full progress of verticillium wilt epidemics in olive. PMID:21255281

  18. A single base pair in the right terminal domain of tomato planta macho viroid is a virulence determinant factor on tomato.

    PubMed

    Li, Rugang; Padmanabhan, Chellappan; Ling, Kai-Shu

    2017-01-01

    Tomato planta macho viroid (TPMVd), including isolates previously designated as Mexican papita viroid (MPVd), causes serious disease on tomatoes in North America. Two predominant variants, sharing 93.8% sequence identity, incited distinct severe (MPVd-S) or mild (MPVd-M) symptoms on tomato. To identify virulence determinant factor, a series of chimeric infectious clones were generated using synthetic DNA approach to progressively replace each structural domain between the two variants. In bioassays on tomato 'Rutgers', three chimeras containing Terminal Left and Pathogenicity (MPVd-H1), Central (MPVd-H2), or Variable (MPVd-H3) of MPVd-S, incited mild to intermediate symptoms. However, a chimera containing Terminal Right (TR) of MPVd-S (MPVd-H4) incited severe symptoms. Only one base-pair mutation in the TR domain between MPVd-M (176U:A185) and MPVd-S (174G:C183) was identified. A reciprocal mutant (MPVd-H5) rendered the chimeric viroid mild on tomato. This single base-pair in the TR domain was determined as the virulence determinant factor for TPMVd. Published by Elsevier Inc.

  19. Silencing of Mythimna separata chitinase genes via oral delivery of in planta-expressed RNAi effectors from a recombinant plant virus.

    PubMed

    Bao, Wenhua; Cao, Budao; Zhang, Yanan; Wuriyanghan, Hada

    2016-11-01

    To evaluate transient expression of RNA interference (RNAi) effectors in Nicotiana benthamiana plants by using recombinant virus vectors and also oral delivery of the effectors for silencing of Mythimna separata endogenous gene expression. Mythimna separata is a serious pest of corn production in China. To evaluate RNAi approaches to target specific RNAs in M. separate, we cloned fragments of the M. separata chitinase sequences into a virus vector in order to produce RNAi effectors during virus infection and replication in plants. When the infected plants were fed to M. separata, expression levels of target MseChi1 and MseChi2 genes were down-regulated by 76 and 45 %, respectively, and sequence-specific siRNAs were detected in recipient insects. RNAi-based silencing of chitinase genes also led to body weight decreases by 43 %. Our research demonstrates target mRNA knockdown and suggests a promising application for controlling of M. separata by in planta expression of RNAi effectors using a recombinant plant virus.

  20. Short-term salinity stress in tobacco plants leads to the onset of animal-like PCD hallmarks in planta in contrast to long-term stress.

    PubMed

    Andronis, Efthimios A; Roubelakis-Angelakis, Kalliopi A

    2010-01-01

    Recent results have identified mitochondria as centers of stress-induced generation of reactive oxygen species in plants. Depolarization of plant mitochondrial membrane during stress results the release of programmed cell death (PCD)-inducing factors in the cytosol in a fashion similar to the onset of animal-like PCD. Herein, we report significant similarities of animal-like PCD and salinity stress-induced plant PCD. Short-term salinity stress (3 h) led to depolarization of the mitochondrial membrane, release of cytochrome c (CYT-c), which was visualized using a contemporary molecular technique, activation of caspase-3 type proteases and the onset of PCD in wild type tobacco plants, Nicotiana tabacum cv. Petit Havana. However, PCD was not manifested during long-term salinity stress (24 h). Interestingly long-term salinity stress led to necrotic-like features, which were accompanied by collapse of respiration, reduction of key components of the respiratory chain, such as CYT-c and alternative oxidase, ATP depletion and high proteolytic activity. The results suggest that salinity stress of tobacco plants in planta leads to the onset of animal-like PCD only during the early stages post-stress, while long-term stress leads to necrotic-like features.

  1. Increased levels of glycerol-3-phosphate lead to a stimulation of flux into triacylglycerol synthesis after supplying glycerol to developing seeds of Brassica napus L. in planta.

    PubMed

    Vigeolas, Helene; Geigenberger, Peter

    2004-09-01

    Glycerol-3-phosphate (glycerol-3P) is a primary substrate for triacylglycerol synthesis. In the present study, changes in the levels of glycerol-3P during rape (Brassica napus L.) seed development and the influence of manipulating glycerol-3P levels on triacylglycerol synthesis were investigated. (i) Glycerol-3P levels were high in young seeds and decreased during seed development at 30 and 40 days after flowering (DAF), when lipid accumulation was maximal. (ii) To manipulate glycerol-3P levels in planta, various concentrations of glycerol were injected directly into 30-DAF seeds, which remained otherwise intact within their siliques and attached to the plant. Injection of 0-10 nmol glycerol led to a progressive increase in seed glycerol-3P levels within 28 h. (iii). Increased levels of glycerol-3P were accompanied by an increase in the flux of injected [14C]sucrose into total lipids and triacylglycerol, whereas fluxes to organic acids, amino acids, starch, protein and cell walls were not affected. (iv) When [14C]acetate was injected into seeds, label incorporation into total lipids and triacylglycerol increased progressively with increasing glycerol-3P levels. (v) There was a strong correlation between the level of glycerol-3P and the incorporation of injected [14C]acetate and [14C]sucrose into triacylglycerol. (v) The results provide evidence that the prevailing levels of glycerol-3P co-limit triacylglycerol synthesis in developing rape seeds.

  2. [Anogenital Warts in a Major Venereology Clinic: Centro de Saúde da Lapa - Lisbon, 2008 to 2014].

    PubMed

    Gameiro, Ana; Alves, João; Santo, Irene; Azevedo, Jacinta

    2016-02-01

    Introdução: O vírus do papiloma humano é responsável pela infeção sexualmente transmissível mais comum, podendo manifestar--se por um conjunto amplo de doenças, nomeadamente condilomas anogenitais, papilomatose laríngea recorrente, e neoplasias da região anogenital, colo do útero e orofaringe. Estima-se que os condilomas anogenitais afetem 1% da população sexualmente ativa, causados em cerca de 90% pelos genótipos 6 e 11. Material e Métodos: Identificámos os doentes com primeiro diagnóstico de condilomas anogenitais da consulta de doenças sexualmente transmissíveis do Centro de Saúde da Lapa, entre janeiro de 2008 e dezembro de 2014, e caracterizámos os doentes por sexo, orientação sexual, idade, localização das lesões, e número de parceiros nos últimos seis meses. Resultados: Foram identificados 902 indivíduos com primeiro diagnóstico de condilomas anogenitais. Observámos uma diminuição significativa de novos casos nas mulheres com < 19 anos (r = -0,848; p = 0,016), e uma diminuição sem significado estatístico nos homens com < 19 anos e nas mulheres entre os 20 - 24 anos. Discussão: Em outubro de 2008, a vacina quadrivalente (genótipos 6, 11, 16, 18) foi introduzida no plano nacional de vacinação de Portugal, abrangendo as adolescentes com 13 anos, com um catch-up para as de 17 anos. A diminuição de primeiros diagnósticos de condilomas anogenitais nas mulheres, deve-se provavelmente à sua vacinação antes de iniciarem a vida sexual. Conclusão: Este estudo reforça a import'ncia do programa nacional de vacinação para o vírus do papiloma humano.

  3. The 32 kDa subunit of replication protein A (RPA) participates in the DNA replication of Mung bean yellow mosaic India virus (MYMIV) by interacting with the viral Rep protein

    PubMed Central

    Singh, Dharmendra Kumar; Islam, Mohammad Nurul; Choudhury, Nirupam Roy; Karjee, Sumona; Mukherjee, Sunil Kumar

    2007-01-01

    Mung bean yellow mosaic India virus (MYMIV) is a member of genus begomoviridae and its genome comprises of bipartite (two components, namely DNA-A and DNA-B), single-stranded, circular DNA of about 2.7 kb. During rolling circle replication (RCR) of the DNA, the stability of the genome and maintenance of the stem–loop structure of the replication origin is crucial. Hence the role of host single-stranded DNA-binding protein, Replication protein A (RPA), in the RCR of MYMIV was examined. Two RPA subunits, namely the RPA70 kDa and RPA32 kDa, were isolated from pea and their roles were validated in a yeast system in which MYMIV DNA replication has been modelled. Here, we present evidences that only the RPA32 kDa subunit directly interacted with the carboxy terminus of MYMIV-Rep both in vitro as well as in yeast two-hybrid system. RPA32 modulated the functions of Rep by enhancing its ATPase and down regulating its nicking and closing activities. The possible role of these modulations in the context of viral DNA replication has been discussed. Finally, we showed the positive involvement of RPA32 in transient replication of the plasmid DNA bearing MYMIV replication origin using an in planta based assay. PMID:17182628

  4. The 32 kDa subunit of replication protein A (RPA) participates in the DNA replication of Mung bean yellow mosaic India virus (MYMIV) by interacting with the viral Rep protein.

    PubMed

    Singh, Dharmendra Kumar; Islam, Mohammad Nurul; Choudhury, Nirupam Roy; Karjee, Sumona; Mukherjee, Sunil Kumar

    2007-01-01

    Mung bean yellow mosaic India virus (MYMIV) is a member of genus begomoviridae and its genome comprises of bipartite (two components, namely DNA-A and DNA-B), single-stranded, circular DNA of about 2.7 kb. During rolling circle replication (RCR) of the DNA, the stability of the genome and maintenance of the stem-loop structure of the replication origin is crucial. Hence the role of host single-stranded DNA-binding protein, Replication protein A (RPA), in the RCR of MYMIV was examined. Two RPA subunits, namely the RPA70 kDa and RPA32 kDa, were isolated from pea and their roles were validated in a yeast system in which MYMIV DNA replication has been modelled. Here, we present evidences that only the RPA32 kDa subunit directly interacted with the carboxy terminus of MYMIV-Rep both in vitro as well as in yeast two-hybrid system. RPA32 modulated the functions of Rep by enhancing its ATPase and down regulating its nicking and closing activities. The possible role of these modulations in the context of viral DNA replication has been discussed. Finally, we showed the positive involvement of RPA32 in transient replication of the plasmid DNA bearing MYMIV replication origin using an in planta based assay.

  5. A distância e o conteúdo estelar da região HII gigante G333.1-0.4 - vínculos para a taxa de formação estelar da galáxia

    NASA Astrophysics Data System (ADS)

    Figuerêdo, E.; Damineli, A.; Blum, R.; Conti, P.

    2003-08-01

    Neste trabalho apresentamos imagens de alta resolução angular da região HII gigante G333.1-0.4 obtidas através dos filtros J, H e K no telescópio de 4-m do CTIO. Este trabalho faz parte de um estudo de regiões HII gigantes no infravermelho próximo que tem por objetivo estudar a natureza da formação de estrelas massivas e traçar a estrutura espiral de nossa galáxia. Nossa determinação da distância é baseada no método da paralaxe espectroscópica de estrelas OB localizadas na seqüência principal de idade zero (ZAMS) do Diagrama HR. No caso de G333.1-0.4, a magnitude aparente das estrelas localizadas na ZAMS indica que a distância não pode ser maior do que o limite inferior determinado por técnica rádio (2,8 kpc). Resultados semelhantes foram encontrados para regiões estudadas anteriormente, reforçando a idéia de que a taxa de formação estelar na Via Láctea é menor do que o determinado a partir de dados rádio. Nossos resultados mais recentes sobre o conteúdo estelar de G333.1-0.4 revelaram vários objetos que possuem cores bastante avermelhadas (H-K > 2,0). Nós identificamos estes objetos usando os diagramas cor-cor e cor-magnitude dos aglomerados. Estes objetos apresentam um forte excesso em emissão na banda K e possivelmente se tratam de estrelas do tipo OB envolvidas por um disco/envelope circumestelar espesso. O estudo da função de massa inical desta região, em conjunto com resultados de nossos trabalhos anteriores, aponta para uma IMF independente da posição galática. A contagem de estrelas nos fornece um valor para o número de fótons no contínuo de Lyman que corrobora com a afirmação de que G333.1-0.4 se encontra mais próxima da menor distância determinada por rádio.

  6. Abundâncias de oxigênio e enxofre nas estrelas de tipo solar da vizinhança solar

    NASA Astrophysics Data System (ADS)

    Requeijo, F.; Porto de Mello, G. F.

    2003-08-01

    Alguns resultados sugerem que o Sol seja 58% mais abundante em oxigênio que o meio interestelar local. Esta anomalia parece estender-se para o carbono e o criptônio. Entre as possíveis explicações deste fenômeno estão: uma supernova de tipo II que tenha enriquecido a nebulosa protosolar, tornando-a superabundante em oxigênio; um episódio de infall de material pobre em metais sobre o disco Galático, diluindo o meio interestelar local ou uma migração dinâmica do Sol de uma órbita mais interna da Galáxia para sua posição atual. A escolha entre estes cenários exige o conhecimento preciso da abundância solar em relação às anãs G da vizinhança. Neste contexto, o oxigênio e enxofre, são elementos-chave por serem ambos produzidos pelas supernovas de tipo II, devendo portanto possuir o mesmo padrão de abundância. Este projeto visa esclarecer qual a posição do Sol na distribuição local de abundâncias de enxofre e oxigênio para uma amostra de estrelas de tipo solar com idades e metalicidades bem conhecidas. Para tal, analisamos espectros de alta resolução e alta relação sinal-ruído nas regiões espectrais de ll 6300, 7774 (O) e l8695 (S). Para o enxofre encontramos que o Sol parece ser uma estrela típica dentre as da vizinhança, e que este elemento não mostra a sobreabundância para baixas metalicidades, já bem estabelecida para o oxigênio. Discutimos as abundâncias do enxofre no contexto da Evolução Química da Galáxia. Apresentamos resultados preliminares muito precisos para a linha proibida do oxigênio l6300 e comparamos estes com os obtidos para o tripleto em l7774. Quantificamos os efeitos não-ETL presentes no tripleto em função dos parâmetros atmosféricos estelares.

  7. The Supernova Impostor SN 2010da

    NASA Astrophysics Data System (ADS)

    Binder, Breanna A.; Williams, Benjamin F.; Kong, Albert K. H.; Plucinsky, Paul P.; Gaetz, Terrance J.; Skillman, Evan D.; Dolphin, Andrew E.

    2016-01-01

    Supernova impostors are optical transients that, despite being assigned a supernova designation, do not signal the death of a massive star or accreting white dwarf. Instead, many impostors are thought to be major eruptions from luminous blue variables. Although the physical cause of these eruptions is still debated, tidal interactions from a binary companion has recently gained traction as a possible explanation for observations of some supernova impostors. In this talk, I will discuss the particularly interesting impostor SN 2010da, which exhibits high-luminosity, variable X-ray emission. The X-ray emission is consistent with accretion onto a neutron star, making SN 2010da a likely high mass X-ray binary in addition to a supernova impostor. SN 2010da is a unique laboratory for understanding both binary interactions as drivers of massive star eruptions and the evolutionary processes that create high mass X-ray binaries.

  8. Boosting in planta production of antigens derived from the porcine reproductive and respiratory syndrome virus (PRRSV) and subsequent evaluation of their immunogenicity.

    PubMed

    Piron, Robin; De Koker, Stefaan; De Paepe, Annelies; Goossens, Julie; Grooten, Johan; Nauwynck, Hans; Depicker, Ann

    2014-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is a disease of swine, caused by an arterivirus, the PRRS virus (PRRSV). This virus infects pigs worldwide and causes huge economic losses. Due to genetic drift, current vaccines are losing their power. Adaptable vaccines could provide a solution to this problem. This study aims at producing in planta a set of antigens derived from the PRRSV glycoproteins (GPs) to be included in a subunit vaccine. We selected the GP3, GP4 and GP5 and optimized these for production in an Arabidopsis seed platform by removing transmembrane domains (Tm) and/or adding stabilizing protein domains, such as the green fluorescent protein (GFP) and immunoglobulin (IgG) 'Fragment crystallizable' (Fc) chains. Accumulation of the GPs with and without Tm was low, reaching no more than 0.10% of total soluble protein (TSP) in homozygous seed. However, addition of stabilizing domains boosted accumulation up to a maximum of 2.74% of TSP when GFP was used, and albeit less effectively, also the Fc chains of the porcine IgG3 and murine IgG2a increased antigen accumulation, to 0.96% and 1.81% of TSP respectively, while the murine IgG3 Fc chain did not. Antigens with Tm were less susceptible to these manipulations to increase yield. All antigens were produced in the endoplasmic reticulum and accordingly, they carried high-mannose N-glycans. The immunogenicity of several of those antigens was assessed and we show that vaccination with purified antigens did elicit the production of antibodies with virus neutralizing activity in mice but not in pigs.

  9. Boosting In Planta Production of Antigens Derived from the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Subsequent Evaluation of Their Immunogenicity

    PubMed Central

    Piron, Robin; De Koker, Stefaan; De Paepe, Annelies; Goossens, Julie; Grooten, Johan; Nauwynck, Hans; Depicker, Ann

    2014-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is a disease of swine, caused by an arterivirus, the PRRS virus (PRRSV). This virus infects pigs worldwide and causes huge economic losses. Due to genetic drift, current vaccines are losing their power. Adaptable vaccines could provide a solution to this problem. This study aims at producing in planta a set of antigens derived from the PRRSV glycoproteins (GPs) to be included in a subunit vaccine. We selected the GP3, GP4 and GP5 and optimized these for production in an Arabidopsis seed platform by removing transmembrane domains (Tm) and/or adding stabilizing protein domains, such as the green fluorescent protein (GFP) and immunoglobulin (IgG) ‘Fragment crystallizable’ (Fc) chains. Accumulation of the GPs with and without Tm was low, reaching no more than 0.10% of total soluble protein (TSP) in homozygous seed. However, addition of stabilizing domains boosted accumulation up to a maximum of 2.74% of TSP when GFP was used, and albeit less effectively, also the Fc chains of the porcine IgG3 and murine IgG2a increased antigen accumulation, to 0.96% and 1.81% of TSP respectively, while the murine IgG3 Fc chain did not. Antigens with Tm were less susceptible to these manipulations to increase yield. All antigens were produced in the endoplasmic reticulum and accordingly, they carried high-mannose N-glycans. The immunogenicity of several of those antigens was assessed and we show that vaccination with purified antigens did elicit the production of antibodies with virus neutralizing activity in mice but not in pigs. PMID:24614617

  10. Endogenous silencing of Puccinia triticina pathogenicity genes through in planta-expressed sequences leads to the suppression of rust diseases on wheat.

    PubMed

    Panwar, Vinay; McCallum, Brent; Bakkeren, Guus

    2013-02-01

    Rust fungi are destructive plant pathogens. The draft genomes of several wheat-infecting species have been released and potential pathogenicity genes identified through comparative analyses to fungal pathogens that are amenable to genetic manipulation. Functional gene analysis tools are needed to understand the infection process of these obligate parasites and to confirm whether predicted pathogenicity genes could become targets for disease control. We have modified an Agrobacterium tumefaciens-mediated in planta-induced transient gene silencing (PITGS) assay for use in Triticum spp. (wheat), and used this assay to target predicted wheat leaf rust fungus, Puccinia triticina (Pt) pathogenicity genes, a MAP kinase (PtMAPK1), a cyclophilin (PtCYC1) and calcineurin B (PtCNB), to analyze their roles in disease. Agroinfiltration effectively delivered hairpin silencing constructs in wheat, leading to the generation of fungal gene-specific siRNA molecules in infiltrated leaves, and resulting in up to 70% reduction in transcription of the endogenous target genes in superinfected Pt. In vivo silencing caused severe disease suppression, compromising fungal growth and sporulation, as viewed by confocal microscopy and measured by reductions in fungal biomass and emergence of uredinia. Interestingly, using the same gene constructs, suppression of infection by Puccinia graminis and Puccinia striiformis was also achieved. Our results show that A. tumefaciens-mediated PITGS can be used as a reverse-genetics tool to discover gene function in rust fungi. This proof-of-concept study indicates that the targeted fungal transcripts might be important in pathogenesis, and could potentially be used as promising targets for developing RNA interference-based resistance against rust fungi. © 2012 The Authors The Plant Journal © 2012 Blackwell Publishing Ltd.

  11. Virulence and in planta movement of Xanthomonas hortorum pv. pelargonii are affected by the diffusible signal factor (DSF)-dependent quorum sensing system.

    PubMed

    Barel, Victoria; Chalupowicz, Laura; Barash, Isaac; Sharabani, Galit; Reuven, Michal; Dror, Orit; Burdman, Saul; Manulis-Sasson, Shulamit

    2015-09-01

    Xanthomonas hortorum pv. pelargonii (Xhp), the causal agent of bacterial blight in pelargonium, is the most threatening bacterial disease of this ornamental worldwide. To gain an insight into the regulation of virulence in Xhp, we have disrupted the quorum sensing (QS) genes, which mediate the biosynthesis and sensing of the diffusible signal factor (DSF). Mutations in rpfF (encoding the DSF synthase) and rpfC (encoding the histidine sensor kinase of the two-component system RfpC/RpfG) and overexpression of rpfF showed a significant reduction in incidence and severity of the disease on pelargonium. Confocal laser scanning microscopy images of inoculated plants with a green fluorescent protein (GFP)-labelled wild-type strain showed that the pathogen is homogeneously dispersed in the lumen of xylem vessels, reaching the apex and invading the intercellular spaces of the leaf mesophyll tissue within 21 days. In contrast, the rpfF and rpfC knockout mutants, as well as the rpfF-overexpressing strain, remained confined to the vicinity of the inoculation site. The rpfF and rpfC mutants formed large incoherent aggregates in the xylem vessels that might interfere with upward movement of the bacterium within the plant. Both mutants also formed extended aggregates under in vitro conditions, whereas the wild-type strain formed microcolonies. Expression levels of putative virulence genes in planta were substantially reduced within 48 h after inoculation with the QS mutants when compared with the wild-type. The results presented indicate that an optimal DSF concentration is crucial for successful colonization and virulence of Xhp in pelargonium. © 2014 BSPP AND JOHN WILEY & SONS LTD.

  12. RiPEIP1, a gene from the arbuscular mycorrhizal fungus Rhizophagus irregularis, is preferentially expressed in planta and may be involved in root colonization.

    PubMed

    Fiorilli, Valentina; Belmondo, Simone; Khouja, Hassine Radhouane; Abbà, Simona; Faccio, Antonella; Daghino, Stefania; Lanfranco, Luisa

    2016-08-01

    Transcriptomics and genomics data recently obtained from the arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis have offered new opportunities to decipher the contribution of the fungal partner to the establishment of the symbiotic association. The large number of genes which do not show similarity to known proteins witnesses the uniqueness of this group of plant-associated fungi. In this work, we characterize a gene that was called RiPEIP1 (Preferentially Expressed In Planta). Its expression is strongly induced in the intraradical phase, including arbuscules, and follows the expression profile of the Medicago truncatula phosphate transporter MtPT4, a molecular marker of a functional symbiosis. Indeed, mtpt4 mutant plants, which exhibit low mycorrhizal colonization and an accelerated arbuscule turnover, also show a reduced RiPEIP1 mRNA abundance. To further characterize RiPEIP1, in the absence of genetic transformation protocols for AM fungi, we took advantage of two different fungal heterologous systems. When expressed as a GFP fusion in yeast cells, RiPEIP1 localizes in the endomembrane system, in particular to the endoplasmic reticulum, which is consistent with the in silico prediction of four transmembrane domains. We then generated RiPEIP1-expressing strains of the fungus Oidiodendron maius, ericoid endomycorrhizal fungus for which transformation protocols are available. Roots of Vaccinium myrtillus colonized by RiPEIP1-expressing transgenic strains showed a higher mycorrhization level compared to roots colonized by the O. maius wild-type strain, suggesting that RiPEIP1 may regulate the root colonization process.

  13. The CRISPR/Cas system can be used as nuclease for in planta gene targeting and as paired nickases for directed mutagenesis in Arabidopsis resulting in heritable progeny.

    PubMed

    Schiml, Simon; Fauser, Friedrich; Puchta, Holger

    2014-12-01

    The CRISPR/Cas nuclease is becoming a major tool for targeted mutagenesis in eukaryotes by inducing double-strand breaks (DSBs) at pre-selected genomic sites that are repaired by non-homologous end joining (NHEJ) in an error-prone way. In plants, it could be demonstrated that the Cas9 nuclease is able to induce heritable mutations in Arabidopsis thaliana and rice. Gene targeting (GT) by homologous recombination (HR) can also be induced by DSBs. Using a natural nuclease and marker genes, we previously developed an in planta GT strategy in which both a targeting vector and targeting locus are activated simultaneously via DSB induction during plant development. Here, we demonstrate that this strategy can be used for natural genes by CRISPR/Cas-mediated DSB induction. We were able to integrate a resistance cassette into the ADH1 locus of A. thaliana via HR. Heritable events were identified using a PCR-based genotyping approach, characterised by Southern blotting and confirmed on the sequence level. A major concern is the specificity of the CRISPR/Cas nucleases. Off-target effects might be avoided using two adjacent sgRNA target sequences to guide the Cas9 nickase to each of the two DNA strands, resulting in the formation of a DSB. By amplicon deep sequencing, we demonstrate that this Cas9 paired nickase strategy has a mutagenic potential comparable with that of the nuclease, while the resulting mutations are mostly deletions. We also demonstrate the stable inheritance of such mutations in A. thaliana. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  14. In Planta Expression Screens of Phytophthora infestans RXLR Effectors Reveal Diverse Phenotypes, Including Activation of the Solanum bulbocastanum Disease Resistance Protein Rpi-blb2[W

    PubMed Central

    Oh, Sang-Keun; Young, Carolyn; Lee, Minkyoung; Oliva, Ricardo; Bozkurt, Tolga O.; Cano, Liliana M.; Win, Joe; Bos, Jorunn I.B.; Liu, Hsin-Yin; van Damme, Mireille; Morgan, William; Choi, Doil; Van der Vossen, Edwin A.G.; Vleeshouwers, Vivianne G.A.A.; Kamoun, Sophien

    2009-01-01

    The Irish potato famine pathogen Phytophthora infestans is predicted to secrete hundreds of effector proteins. To address the challenge of assigning biological functions to computationally predicted effector genes, we combined allele mining with high-throughput in planta expression. We developed a library of 62 infection-ready P. infestans RXLR effector clones, obtained using primer pairs corresponding to 32 genes and assigned activities to several of these genes. This approach revealed that 16 of the 62 examined effectors cause phenotypes when expressed inside plant cells. Besides the well-studied AVR3a effector, two additional effectors, PexRD8 and PexRD3645-1, suppressed the hypersensitive cell death triggered by the elicitin INF1, another secreted protein of P. infestans. One effector, PexRD2, promoted cell death in Nicotiana benthamiana and other solanaceous plants. Finally, two families of effectors induced hypersensitive cell death specifically in the presence of the Solanum bulbocastanum late blight resistance genes Rpi-blb1 and Rpi-blb2, thereby exhibiting the activities expected for Avrblb1 and Avrblb2. The AVRblb2 family was then studied in more detail and found to be highly variable and under diversifying selection in P. infestans. Structure-function experiments indicated that a 34–amino acid region in the C-terminal half of AVRblb2 is sufficient for triggering Rpi-blb2 hypersensitivity and that a single positively selected AVRblb2 residue is critical for recognition by Rpi-blb2. PMID:19794118

  15. Hidden sketches by Leonardo da Vinci revealed

    NASA Astrophysics Data System (ADS)

    Dumé, Belle

    2009-02-01

    Three drawings on the back of Leonardo da Vinci's The Virgin and Child with St Anne (circa 1508) have been discovered by researchers led by Michel Menu from the Centre de Recherche et de Restauration des Musées de France (C2RMF) and the Louvre Museum in Paris.

  16. DA white dwarfs in the Kepler field

    NASA Astrophysics Data System (ADS)

    Doyle, T. F.; Howell, S. B.; Petit, V.; Lépine, S.

    2017-01-01

    We present 16 new, and confirm 7 previously identified, DA white dwarfs in the Kepler field through ground-based spectroscopy with the Hale 200″, Kitt Peak 4-m, and Bok 2.3-m telescopes. Using atmospheric models, we determine their effective temperatures and surface gravities to constrain their position with respect to the ZZ Ceti (DA pulsator) instability strip, and look for the presence or absence of pulsation with Kepler's unprecedented photometry. Our results are as follows. (i) From our measurements of temperature and surface gravity, 12 of the 23 DA white dwarfs from this work fall well outside of the instability strip. The Kepler photometry available for 11 of these WDs allows us to confirm that none are pulsating. One of these 11 happens to be a presumed binary, KIC 11604781, with a period of ˜5 d. (ii) The remaining 11 DA white dwarfs are instability strip candidates, potentially falling within the current, empirical instability strip, after accounting for uncertainties. These WDs will help constrain the strip's location further, as eight are near the blue edge and three are near the red edge of the instability strip. Four of these WDs do not have Kepler photometry, so ground-based photometry is needed to determine the pulsation nature of these white dwarfs. The remaining seven have Kepler photometry available, but do not show any periodicity on typical WD pulsation time-scales.

  17. How to Think Like Leonardo da Vinci

    ERIC Educational Resources Information Center

    Caouette, Ralph

    2008-01-01

    To be effective and relevant in twenty-first-century learning, art needs to be more inclusive. In this article, the author discusses how teachers can find a good example in Leonardo da Vinci for building an art program. His art, design, and curiosity are the perfect foundation for any art program, at any level. (Contains 3 resources and 3 online…

  18. How to Think Like Leonardo da Vinci

    ERIC Educational Resources Information Center

    Caouette, Ralph

    2008-01-01

    To be effective and relevant in twenty-first-century learning, art needs to be more inclusive. In this article, the author discusses how teachers can find a good example in Leonardo da Vinci for building an art program. His art, design, and curiosity are the perfect foundation for any art program, at any level. (Contains 3 resources and 3 online…

  19. VerSeDa: vertebrate secretome database

    PubMed Central

    Cortazar, Ana R.; Oguiza, José A.

    2017-01-01

    Based on the current tools, de novo secretome (full set of proteins secreted by an organism) prediction is a time consuming bioinformatic task that requires a multifactorial analysis in order to obtain reliable in silico predictions. Hence, to accelerate this process and offer researchers a reliable repository where secretome information can be obtained for vertebrates and model organisms, we have developed VerSeDa (Vertebrate Secretome Database). This freely available database stores information about proteins that are predicted to be secreted through the classical and non-classical mechanisms, for the wide range of vertebrate species deposited at the NCBI, UCSC and ENSEMBL sites. To our knowledge, VerSeDa is the only state-of-the-art database designed to store secretome data from multiple vertebrate genomes, thus, saving an important amount of time spent in the prediction of protein features that can be retrieved from this repository directly. Database URL: VerSeDa is freely available at http://genomics.cicbiogune.es/VerSeDa/index.php PMID:28365718

  20. VerSeDa: vertebrate secretome database.

    PubMed

    Cortazar, Ana R; Oguiza, José A; Aransay, Ana M; Lavín, José L

    2017-01-01

    Based on the current tools, de novo secretome (full set of proteins secreted by an organism) prediction is a time consuming bioinformatic task that requires a multifactorial analysis in order to obtain reliable in silico predictions. Hence, to accelerate this process and offer researchers a reliable repository where secretome information can be obtained for vertebrates and model organisms, we have developed VerSeDa (Vertebrate Secretome Database). This freely available database stores information about proteins that are predicted to be secreted through the classical and non-classical mechanisms, for the wide range of vertebrate species deposited at the NCBI, UCSC and ENSEMBL sites. To our knowledge, VerSeDa is the only state-of-the-art database designed to store secretome data from multiple vertebrate genomes, thus, saving an important amount of time spent in the prediction of protein features that can be retrieved from this repository directly. VerSeDa is freely available at http://genomics.cicbiogune.es/VerSeDa/index.php.

  1. The local effect of octreotide on mechanical pain sensitivity is more sensitive in DA rats than DA.1U rats.

    PubMed

    Yao, Fan-Rong; Wang, Hui-Sheng; Guo, Yuan; Zhao, Yan

    2016-02-01

    A recent study by the authors indicated that major histocompatibility complex (MHC) genes are associated with the differences in basal pain sensitivity and in formalin model between Dark-Agouti (DA) and novel congenic DA.1U rats, which have the same genetic background as DA rats except for the u alleles of MHC. The objective of the present study is to investigate whether there is a difference in the pristane-induced arthritis (PIA) model and local analgesic effect of octreotide (OCT) between DA and DA.1U rats. The hindpaw mechanical withdrawal threshold (MWT) and heat withdrawal latency (HWL) were observed. The C unit firings of the tibial nerve evoked by non-noxious and noxious toe movements were recorded by electrophysiological methods in normal and PIA models in DA and DA.1U rats before and after local OCT administration. The expression of somatostatin receptor 2A (SSTR2A) was observed by immunohistochemistry. The results demonstrate that DA rats have a higher mechanical sensitivity than DA.1U rats after PIA. Local OCT administration significantly elevated MWT in DA rats under normal and PIA sate, but not in DA.1U rats. The electrophysiological experiments showed OCT significantly attenuated the firings of C units evoked by non-noxious and noxious stimulation in DA rats more than those in DA.1U rats both in normal and PIA states. In addition, the expression of SSTR2A in the dorsal horn of the spinal cord was significantly higher in DA than in DA.1U rats. All of the findings suggest a higher local analgesic effect of OCT in DA rats than DA.1U rats, which might be associated with the MHC genes.

  2. A Day in the Life at DaVita Academy

    ERIC Educational Resources Information Center

    Weinstein, Margery

    2010-01-01

    When a company name means "giving life," the bar for learning and development programs is held high. In this article, the author describes what it takes to graduate from DaVita Academy, the soft skills training program dialysis services company DaVita offers all its employees. DaVita's chief executive officer, Kent Thiry, states that the Academy…

  3. A Day in the Life at DaVita Academy

    ERIC Educational Resources Information Center

    Weinstein, Margery

    2010-01-01

    When a company name means "giving life," the bar for learning and development programs is held high. In this article, the author describes what it takes to graduate from DaVita Academy, the soft skills training program dialysis services company DaVita offers all its employees. DaVita's chief executive officer, Kent Thiry, states that the Academy…

  4. 32 CFR 516.25 - DA Form 4.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 3 2010-07-01 2010-07-01 true DA Form 4. 516.25 Section 516.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY AID OF CIVIL AUTHORITIES AND PUBLIC RELATIONS LITIGATION Reporting Legal Proceedings to HQDA § 516.25 DA Form 4. (a) General. The DA Form 4 (See figure...

  5. Tdp studies and tests for C. A. Energia Electrica de Venezuela (enelven) at planta ramon laguna, units RL-17 and RL-10. Volume 2. Unit RL-10 boiler condition assessment report. Export trade information

    SciTech Connect

    Not Available

    1991-03-28

    The study, conducted by Babcock and Wilcox, was funded by the U.S. Trade and Development agency on behalf of Enelven. In order to maximize generated power output and minimize operating costs at Planta Ramon Laguna, tests were done to evaluate the condition of equipment at the plant. In order to identify any damage and determine the operating output of each unit, assessments were done of the furnaces, boilers, generators and boiler feed pumps being used in the plant. The report presents the results of these tests. This is the second of three volumes and it includes the following section: (1) Condition Assessment of Unit RL-10 Boiler.

  6. Characterization of two catalase-peroxidase encoding genes in Fusarium verticillioides reveals differential responses to in vitro versus in planta oxidative challenges.

    PubMed

    Gao, Shan; Gold, Scott E; Glenn, Anthony E

    2017-08-12

    Catalase/peroxidases (KatGs) are a superfamily of reactive oxygen species (ROS)-degrading enzymes believed to be horizontally acquired by ancient Ascomycota from bacteria. Subsequent gene duplication resulted in two KatG paralogs in ascomycetes: the widely distributed intracellular KatG1 group, and the phytopathogen-dominated extracellular KatG2 group. To functionally characterize FvCP01 (KatG1) and FvCP02 (KatG2) in the maize pathogen Fusarium verticillioides, single and double gene deletion mutants were examined in response to hydrogen peroxide (H2 O2 ). Both ΔFvCP01 and ΔFvCP02 were more sensitive to H2 O2 than the wild type in vitro, although their sensitivity differed depending on the type of inoculum. Inoculations using mycelial agar plugs demonstrated an additive effect of the mutants, with the ΔFvCP01/ΔFvCP02 double deletion being the most sensitive to H2 O2 . Conidia in general were much more sensitive to H2 O2 than agar plugs, and conidial inoculations indicated FvCP01 conferred more H2 O2 tolerance than FvCP02. Transcriptional analysis showed induction of FvCP01 but decreased expression of FvCP02 in both mycelia and spores in wild type after H2 O2 exposure, but this trend was reversed when the fungus was grown on germinating maize seeds. This interaction with the plant increased expression of FvCP02 but not FvCP01, indicating FvCP02 may be responsive to plant-derived H2 O2 . Yet, FvCP01 was induced >3-fold in the ΔFvCP02 mutant grown on germlings, suggesting FvCP01 can compensate for the loss of FvCP02. Given the differential responses of these two F. verticillioides genes to in vitro versus in planta challenges, a model is proposed to illustrate the differing roles of FvCP01 and FvCP02 in protective responses against H2 O2 -derived oxidative stress. This article is protected by copyright. All rights reserved. © 2017 BSPP and John Wiley & Sons Ltd.

  7. Genomic homologous recombination in planta.

    PubMed Central

    Gal, S; Pisan, B; Hohn, T; Grimsley, N; Hohn, B

    1991-01-01

    A system for monitoring intrachromosomal homologous recombination in whole plants is described. A multimer of cauliflower mosaic virus (CaMV) sequences, arranged such that CaMV could only be produced by recombination, was integrated into Brassica napus nuclear DNA. This set-up allowed scoring of recombination events by the appearance of viral symptoms. The repeated homologous regions were derived from two different strains of CaMV so that different recombinant viruses (i.e. different recombination events) could be distinguished. In most of the transgenic plants, a single major virus species was detected. About half of the transgenic plants contained viruses of the same type, suggesting a hotspot for recombination. The remainder of the plants contained viruses with cross-over sites distributed throughout the rest of the homologous sequence. Sequence analysis of two recombinant molecules suggest that mismatch repair is linked to the recombination process. Images PMID:2026150

  8. Homogeneous generation of iDA neurons with high similarity to bona fide DA neurons using a drug inducible system.

    PubMed

    Park, Hanseul; Kim, Hongwon; Yoo, Junsang; Lee, Jaekwang; Choi, Hwan; Baek, Soonbong; Lee, C Justin; Kim, Janghwan; Lengner, Christopher J; Sung, Jung-Suk; Kim, Jongpil

    2015-12-01

    Recent work generating induced dopaminergic (iDA) neurons using direct lineage reprogramming potentially provides a novel platform for the study and treatment Parkinson's disease (PD). However, one of the most important issues for iDA-based applications is the degree to which iDA neurons resemble the molecular and functional properties of their endogenous DA neuron counterparts. Here we report that the homogeneity of the reprogramming gene expression system is critical for the generation of iDA neuron cultures that are highly similar to endogenous DA neurons. We employed an inducible system that carries iDA-inducing factors as defined transgenes for direct lineage reprogramming to iDA neurons. This system circumvents the need for viral transduction, enabling a more efficient and reproducible reprogramming process for the generation of genetically homogenous iDA neurons. We showed that this inducible system generates iDA neurons with high similarity to their bona fide in vivo counterparts in comparison to direct infection methods. Thus, our results suggest that homogenous expression of exogenous genes in direct lineage reprogramming is critical for the generation of high quality iDA neuron cultures, making such culture systems a valuable resource for iDA-based drug screening and, ultimately, potential therapeutic intervention in PD. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. PanDA for COMPASS at JINR

    NASA Astrophysics Data System (ADS)

    Petrosyan, A. Sh.

    2016-09-01

    PanDA (Production and Distributed Analysis System) is a workload management system, widely used for data processing at experiments on Large Hadron Collider and others. COMPASS is a high-energy physics experiment at the Super Proton Synchrotron. Data processing for COMPASS runs locally at CERN, on lxbatch, the data itself stored in CASTOR. In 2014 an idea to start running COMPASS production through PanDA arose. Such transformation in experiment's data processing will allow COMPASS community to use not only CERN resources, but also Grid resources worldwide. During the spring and summer of 2015 installation, validation and migration work is being performed at JINR. Details and results of this process are presented in this paper.

  10. [Leonardo da Vinci--a dyslectic genius?].

    PubMed

    Røsstad, Anna

    2002-12-10

    Leonardo da Vinci's texts consist almost exclusively of scientific notes. Working on a book on Leonardo's art, I studied all Leonardo's published texts carefully for any new information. In some prefaces I came to suspect that Leonardo might have suffered from dyslexia. This article considers the question of whether it is possible to find indications of dyslexia in Leonardo's texts and in the accounts of his life.

  11. DA 495: An Aging Pulsar Wind Nebula

    NASA Astrophysics Data System (ADS)

    Kothes, R.; Landecker, T. L.; Reich, W.; Safi-Harb, S.; Arzoumanian, Z.

    2008-11-01

    We present a radio continuum study of the pulsar wind nebula (PWN) DA 495 (G65.7+1.2), including images of total intensity and linear polarization from 408 to 10550 MHz based on the Canadian Galactic Plane Survey and observations with the Effelsberg 100 m Radio Telescope. Removal of flux density contributions from a superimposed H II region and from compact extragalactic sources reveals a break in the spectrum of DA 495 at 1.3 GHz, with a spectral index α = - 0.45 +/- 0.20 below the break and α = - 0.87 +/- 0.10 above it (Sν propto να). The spectral break is more than 3 times lower in frequency than the lowest break detected in any other PWN. The break in the spectrum is likely the result of synchrotron cooling, and DA 495, at an age of ~20,000 yr, may have evolved from an object similar to the Vela X nebula, with a similarly energetic pulsar. We find a magnetic field of ~1.3 mG inside the nebula. After correcting for the resulting high internal rotation measure, the magnetic field structure is quite simple, resembling the inner part of a dipole field projected onto the plane of the sky, although a toroidal component is likely also present. The dipole field axis, which should be parallel to the spin axis of the putative pulsar, lies at an angle of ~50° east of the north celestial pole and is pointing away from us toward the southwest. The upper limit for the radio surface brightness of any shell-type supernova remnant emission around DA 495 is Σ1GHz ~ 5.4 × 10-23 W m-2 Hz-1 sr-1 (assuming a radio spectral index of α = - 0.5), lower than the faintest shell-type remnant known to date.

  12. UBV photometry of hot DA white dwarfs

    NASA Technical Reports Server (NTRS)

    Kidder, K. M.; Holberg, J. B.; Mason, Paul A.

    1991-01-01

    Johnson UBV photometry has been obtained photoelectrically for a set of DA white dwarfs with effective temperatures greater than 20,000 K and for the AM Her type binary HO538 + 608. Most of the white dwarfs lie within existing Einstein IPC or EXOSAT LE soft X-ray fields, therefore they are of interest as potential serendipitous soft X-ray sources. In addition, high dispersion spectroscopy has been used to differentiate seven of these objects to be subdwarfs.

  13. A computational theory of da Vinci stereopsis.

    PubMed

    Tsirlin, Inna; Wilcox, Laurie M; Allison, Robert S

    2014-06-09

    In binocular vision, occlusion of one object by another gives rise to monocular occlusions—regions visible only in one eye. Although binocular disparities cannot be computed for these regions, monocular occlusions can be precisely localized in depth and can induce the perception of illusory occluding surfaces. The phenomenon of depth perception from monocular occlusions, known as da Vinci stereopsis, is intriguing, but its mechanisms are not well understood. We first propose a theory of the mechanisms underlying da Vinci stereopsis that is based on the psychophysical and computational literature on monocular occlusions. It postulates, among other principles, that monocular areas are detected explicitly, and depth from occlusions is calculated based on constraints imposed by occlusion geometry. Next, we describe a biologically inspired computational model based on this theory that successfully reconstructs depth in a large range of stimuli and produces results similar to those described in the psychophysical literature. These results demonstrate that the proposed neural architecture could underpin da Vinci stereopsis and other stereoscopic percepts. © 2014 ARVO.

  14. Leonardo da Vinci's studies of the heart.

    PubMed

    Shoja, Mohammadali M; Agutter, Paul S; Loukas, Marios; Benninger, Brion; Shokouhi, Ghaffar; Namdar, Husain; Ghabili, Kamyar; Khalili, Majid; Tubbs, R Shane

    2013-08-20

    Leonardo da Vinci's detailed drawings are justly celebrated; however, less well known are his accounts of the structures and functions of the organs. In this paper, we focus on his illustrations of the heart, his conjectures about heart and blood vessel function, his experiments on model systems to test those conjectures, and his unprecedented conclusions about the way in which the cardiovascular system operates. In particular, da Vinci seems to have been the first to recognize that the heart is a muscle and that systole is the active phase of the pump. He also seems to have understood the functions of the auricles and pulmonary veins, identified the relationship between the cardiac cycle and the pulse, and explained the hemodynamic mechanism of valve opening and closure. He also described anatomical variations and changes in structure and function that occurred with age. We outline da Vinci's varied career and suggest ways in which his personality, experience, skills and intellectual heritage contributed to these advances in understanding. We also consider his influence on later studies in anatomy and physiology.

  15. The real code of leonardo da vinci.

    PubMed

    Ose, Leiv

    2008-02-01

    Leonardo da Vinci was born in Italy. Among the researchers and scientists, he is favourably known for his remarkable efforts in scientific work. His investigations of atherosclerosis judiciously combine three separate fields of research. In 1506, he finished his masterpiece, painting of Mona Lisa. A careful clinical examination of the famous painting reveals a yellow irregular leather-like spot at the inner end of the left upper eyelid and a soft bumpy well-defined swelling of the dorsum of the right hand beneath the index finger about 3 cm long. This is probably the first case of familial hypercholesterolemia (FH). The FH code of Leonardo da Vinci was given immense consideration by scientists like Carl Muller, who described the xanthomas tuberosum and angina pectoris. On the contrary, Akira Endo searched for microbial metabolites that would inhibit HMG-CoA reductase, the rate-limiting enzyme in the synthesis of cholesterol and finally, Michael Brown and Joseph Goldstein published a remarkable series of elegant and insightful papers in the 70s and 80s. They established that the cellular uptake of low-density lipoprotein (LDL) essentially requires the LDL receptor. this was the real Code of Leonardo da Vinci.

  16. The Real Code of Leonardo da Vinci

    PubMed Central

    Ose, Leiv

    2008-01-01

    Leonardo da Vinci was born in Italy. Among the researchers and scientists, he is favourably known for his remarkable efforts in scientific work. His investigations of atherosclerosis judiciously combine three separate fields of research. In 1506, he finished his masterpiece, painting of Mona Lisa. A careful clinical examination of the famous painting reveals a yellow irregular leather-like spot at the inner end of the left upper eyelid and a soft bumpy well-defined swelling of the dorsum of the right hand beneath the index finger about 3 cm long. This is probably the first case of familial hypercholesterolemia (FH). The FH code of Leonardo da Vinci was given immense consideration by scientists like Carl Muller, who described the xanthomas tuberosum and angina pectoris. On the contrary, Akira Endo searched for microbial metabolites that would inhibit HMG-CoA reductase, the rate-limiting enzyme in the synthesis of cholesterol and finally, Michael Brown and Joseph Goldstein published a remarkable series of elegant and insightful papers in the 70s and 80s. They established that the cellular uptake of low-density lipoprotein (LDL) essentially requires the LDL receptor. In conclusion: this was the real Code of Leonardo da Vinci. PMID:19924278

  17. Use of Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) Spectroscopy in Direct, Nondestructive, and Rapid Assessment of Developmental Cotton Fibers Grown In Planta and in Culture.

    PubMed

    Liu, Yongliang; Kim, Hee-Jin

    2015-08-01

    Cotton fibers are routinely harvested from cotton plants (in planta), and their end-use qualities depend on their development stages. Cotton fibers are also cultured in controlled laboratory environments, so that cotton researchers can investigate many aspects of experimental protocols in cotton breeding programs at reduced expenses. In this work, attenuated total reflection Fourier transform infrared (ATR FT-IR) spectra of cotton fibers grown in planta and in culture were collected to explore the potential of FT-IR technique as a simple, rapid, and direct method for characterizing the fiber development. Complementary to visual inspection of spectral variations, principal component analysis (PCA) of ATR FT-IR spectra revealed the occurrence of phase transition from primary to secondary cell wall synthesis and also the difference of starting the phase transition between two types of fibers. Like PCA observation, three simple algorithms were capable of monitoring the secondary cell wall formation effectively. Interestingly and uniquely, simple algorithms were able to detect the subtle discrepancies in fibers older than 25 days post-anthesis, which was not apparent from PCA results. The observation indicated the feasibility of FT-IR technique in rapid, routine, nondestructive, and direct assessment of fiber development for cotton physiology and breeding applications.

  18. [Agrobacterium-mediated sunflower transformation (Helianthus annuus L.) in vitro and in Planta using strain of LBA4404 harboring binary vector pBi2E with dsRNA-suppressor proline dehydrogenase gene].

    PubMed

    Tishchenko, E N; Komisarenko, A G; Mikhal'skaia, S I; Sergeeva, L E; Adamenko, N I; Morgun, B V; Kochetov, A V

    2014-01-01

    To estimate the efficiency of proline dehydrogenase gene suppression towards increasing of sunflower (Helianthus annuus L.) tolerance level to water deficit and salinity, we employed strain LBA4404 harboring pBi2E with double-stranded RNA-suppressor, which were prepared on basis arabidopsis ProDH1 gene. The techniques of Agrobacterium-mediated transformation in vitro and in planta during fertilization sunflower have been proposed. There was shown the genotype-depended integration of T-DNA in sunflower genome. PCR-analysis showed that ProDH1 presents in genome of inbred lines transformed in planta, as well as in T1- and T2-generations. In trans-genic regenerants the essential accumulation of free L-proline during early stages of in vitro cultivation under normal conditions was shown. There was established the essential accumulation of free proline in transgenic regenerants during cultivation under lethal stress pressure (0.4 M mannitol and 2.0% sea water salts) and its decline upon the recovery period. These data are declared about effectiveness of suppression of sunflower ProDH and gene participation in processes connected with osmotolerance.

  19. Rational design of D-A1-D-A2 conjugated polymers with superior spectral coverage.

    PubMed

    Hedström, Svante; Tao, Qiang; Wang, Ergang; Persson, Petter

    2015-10-28

    The spectral coverage of a light-harvesting polymer largely determines the maximum achievable photocurrent in organic photovoltaics, and therefore constitutes a crucial parameter for improving their performance. The D-A1-D-A2 copolymer motif is a new and promising design strategy for extending the absorption range by incorporating two acceptor units with complementary photoresponses. The fundamental factors that promote an extended absorption are here determined for three prototype D-A1-D-A2 systems through a combination of experimental and computational methods. Systematic quantum chemical calculations are then used to reveal the intrinsic optical properties of ten further D-A1-D-A2 polymer candidates. These investigated polymers are all predicted to exhibit intense primary absorption peaks at 615-954 nm, corresponding to charge-transfer (CT) transitions to the stronger acceptor, as well as secondary absorption features at 444-647 nm that originate from CT transitions to the weaker acceptors. Realization of D-A1-D-A2 polymers with superior spectral coverage is thereby found to depend critically on the spatial and energetic separation between the two distinct acceptor LUMOs. Two promising D-A1-D-A2 copolymer candidates were finally selected for further theoretical and experimental study, and demonstrate superior light-harvesting properties in terms of significantly extended spectral coverage. This demonstrates great potential for enhanced light-harvesting in D-A1-D-A2 polymers via multiple absorption features compared to traditional D-A polymers.

  20. DA{phi}NE beam instrumentation

    SciTech Connect

    Ghigo, A.; Biscari, C.; Coiro, O.; Pirro, G. Di; Drago, A.; Gallo, A.; Marcellini, F.; Mazzitelli, G.; Milardi, C.; Sannibale, F.; Serio, M.; Stecchi, A.; Stella, A.; Vignola, G.; Zobov, M.

    1998-12-10

    DA{phi}NE, the Frascati {phi}-Factory, is now under commissioning. The accelerator complex is composed of a linac, an accumulator-damping ring, and two separate main rings, one for electrons and the other for positrons, with two interaction regions in which the experiments will be placed. In order to achieve the luminosity goal, high performance instrumentation and beam diagnostics have been installed. Some of the relevant beam measurements performed are: beam emittance, transverse and longitudinal dimensions, beam positions and tunes, overlap in the interaction points, and luminosity. An overview of the diagnostic instrumentation of the accelerator complex is given together with measurement examples and discussion of operational experiences.

  1. Polarization-switching D/A converter.

    PubMed

    Sun, Shunming; Kalkur, Thottam S

    2005-05-01

    This paper describes a novel digital-to-analog (D/A) conversion technique, which uses the analog quantity polarization as a D/A conversion medium. It can be implemented by CMOS capacitors or by ferroelectric capacitors, which exhibit strong nonlinearity in charge versus voltage behavior. Because a ferroelectric material inherently has spontaneous polarization and generally has a large dielectric constant, the effective capacitance of a ferroelectric capacitor is much larger than that of a CMOS capacitor of the same size. This ensures less influence of bottom-electrode parasitic capacitance on a ferroelectric capacitor. Furthermore, a data converter based on ferroelectric capacitors possesses the potential nonvolatile memory function owing to ferroelectric hysteresis. Along with the architecture proposed for polarization-switching digital-to-analog converter (PDAC), its circuit implementation is introduced. Described is implementation of two 9-bit bipolar PDACs: one is based on CMOS capacitors and the other on off-chip ferroelectric capacitors. Experimental results are presented for the performance of these two prototypes.

  2. Tdp studies and tests for C. A. Energia Electrica de Venezuela (enelven) at planta ramon laguna, units RL-17 and RL-10. Volume 1. Executive summary, RL-17 test report, and gas conversion proposals. Export trade information

    SciTech Connect

    Not Available

    1991-03-28

    The study, conducted by Babcock and Wilcox, was funded by the U.S. Trade and Development agency on behalf of Enelven. In order to maximize generated power output and minimize operating costs at Planta Ramon Laguna, tests were done to evaluate the condition of equipment at the plant. In order to identify any damage and determine the operating output of each unit, assessments were done of the furnaces, boilers, generators and boiler feed pumps being used in the plant. The report presents the results of these tests. This is the first of three volumes and it is divided into the following sections: (1) Executive Summary; (2) Hydrogen Damage Assessment; (3) RL-17 Gas Conversion Proposal; (4) RL-10 and RL-11 Gas Conversion Proposals.

  3. 32 CFR 643.121 - Private organizations on DA installations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 4 2011-07-01 2011-07-01 false Private organizations on DA installations. 643... (CONTINUED) REAL PROPERTY REAL ESTATE Additional Authority of Commanders § 643.121 Private organizations on DA installations. (a) AR 210-1 defines and classifies private organizations, such as thrift shops...

  4. 32 CFR 516.25 - DA Form 4.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 3 2014-07-01 2014-07-01 false DA Form 4. 516.25 Section 516.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY AID OF CIVIL AUTHORITIES AND PUBLIC... attached to a properly prepared and sealed DA Form 4 are self-authenticating. (See Fed. R. Evid. 902). (b...

  5. 32 CFR 516.25 - DA Form 4.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 3 2012-07-01 2009-07-01 true DA Form 4. 516.25 Section 516.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY AID OF CIVIL AUTHORITIES AND PUBLIC RELATIONS... properly prepared and sealed DA Form 4 are self-authenticating. (See Fed. R. Evid. 902). (b) Preparation at...

  6. DA-6034 Induces [Ca(2+)]i Increase in Epithelial Cells.

    PubMed

    Yang, Yu-Mi; Park, Soonhong; Ji, Hyewon; Kim, Tae-Im; Kim, Eung Kweon; Kang, Kyung Koo; Shin, Dong Min

    2014-04-01

    DA-6034, a eupatilin derivative of flavonoid, has shown potent effects on the protection of gastric mucosa and induced the increases in fluid and glycoprotein secretion in human and rat corneal and conjunctival cells, suggesting that it might be considered as a drug for the treatment of dry eye. However, whether DA-6034 induces Ca(2+) signaling and its underlying mechanism in epithelial cells are not known. In the present study, we investigated the mechanism for actions of DA-6034 in Ca(2+) signaling pathways of the epithelial cells (conjunctival and corneal cells) from human donor eyes and mouse salivary gland epithelial cells. DA-6034 activated Ca(2+)-activated Cl(-) channels (CaCCs) and increased intracellular calcium concentrations ([Ca(2+)]i) in primary cultured human conjunctival cells. DA-6034 also increased [Ca(2+)]i in mouse salivary gland cells and human corneal epithelial cells. [Ca(2+)]i increase of DA-6034 was dependent on the Ca(2+) entry from extracellular and Ca(2+) release from internal Ca(2+) stores. Interestingly, these effects of DA-6034 were related to ryanodine receptors (RyRs) but not phospholipase C/inositol 1,4,5-triphosphate (IP3) pathway and lysosomal Ca(2+) stores. These results suggest that DA-6034 induces Ca(2+) signaling via extracellular Ca(2+) entry and RyRs-sensitive Ca(2+) release from internal Ca(2+) stores in epithelial cells.

  7. The Case: Bunche-Da Vinci Learning Partnership Academy

    ERIC Educational Resources Information Center

    Eisenberg, Nicole; Winters, Lynn; Alkin, Marvin C.

    2005-01-01

    The Bunche-Da Vinci case described in this article presents a situation at Bunche Elementary School that four theorists were asked to address in their evaluation designs (see EJ791771, EJ719772, EJ791773, and EJ792694). The Bunche-Da Vinci Learning Partnership Academy, an elementary school located between an urban port city and a historically…

  8. The Case: Bunche-Da Vinci Learning Partnership Academy

    ERIC Educational Resources Information Center

    Eisenberg, Nicole; Winters, Lynn; Alkin, Marvin C.

    2005-01-01

    The Bunche-Da Vinci case described in this article presents a situation at Bunche Elementary School that four theorists were asked to address in their evaluation designs (see EJ791771, EJ719772, EJ791773, and EJ792694). The Bunche-Da Vinci Learning Partnership Academy, an elementary school located between an urban port city and a historically…

  9. Leonardo da Vinci's contributions to neuroscience.

    PubMed

    Pevsner, Jonathan

    2002-04-01

    Leonardo da Vinci (1452-1519) made far-reaching contributions to many areas of science, technology and art. Leonardo's pioneering research into the brain led him to discoveries in neuroanatomy (such as those of the frontal sinus and meningeal vessels) and neurophysiology (he was the first to pith a frog). His injection of hot wax into the brain of an ox provided a cast of the ventricles, and represents the first known use of a solidifying medium to define the shape and size of an internal body structure. Leonardo developed an original, mechanistic model of sensory physiology. He undertook his research with the broad goal of providing physical explanations of how the brain processes visual and other sensory input, and integrates that information via the soul.

  10. The DA{phi}NE luminosity monitor

    SciTech Connect

    Di Pirro, G.; Drago, A.; Ghigo, A.; Mazzitelli, G.; Preger, M.; Sannibale, F.; Serio, M.; Vignola, G.; Cervelli, F.; Lomtadze, T.

    1998-12-10

    DA{phi}NE, the Frascati {phi}-factory, is an e{sup +}/e{sup -} collider with 2 interaction points (IPs). The center of mass energy is 1020 MeV and the design luminosity 4.2x10{sup 30} cm{sup -2} s{sup -1} in single bunch mode and 5x10{sup 32} cm{sup -2} s{sup -1} in multibunch mode. Between the possible electromagnetic reactions at the interaction point, single bremsstrahlung (SB) has been selected for the luminosity measurement. The SB high counting rate allows real-time monitoring, which is very useful during machine tune-up and moreover the narrow peak of the SB angular distribution makes the counting rate almost independent from the beam position at the IP. A description of the experimental set-up, calibration results and luminosity measurements is presented.

  11. Da Costa's syndrome or neurocirculatory asthenia.

    PubMed Central

    Paul, O

    1987-01-01

    The syndrome variously called Da Costa's syndrome, effort syndrome, neurocirculatory asthenia, etc has been studied for more than 100 years by many distinguished physicians. Originally identified in men in wartime, it has been widely recognised as a common chronic condition in both sexes in civilian life. Although the symptoms may seem to appear after infections and various physical and psychological stresses, neurocirculatory asthenia is most often encountered as a familial disorder that is unrelated to these factors, although they may aggravate an existing tendency. Respiratory complaints (including breathlessness, with and without effort, and smothering sensations) are almost universal, and palpitation, chest discomfort, dizziness and faintness, and fatigue are common. The physical examination is normal. The aetiology is obscure but patients usually have a normal life span. Reassurance and measures to improve physical fitness are helpful. PMID:3314950

  12. 75 FR 52292 - Airworthiness Directives; Diamond Aircraft Industries GmbH Models DA 40 and DA 40F Airplanes

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-25

    ... Directives; Diamond Aircraft Industries GmbH Models DA 40 and DA 40F Airplanes AGENCY: Federal Aviation...: We propose to adopt a new airworthiness directive (AD) for all Diamond Aircraft Industries GmbH... Federal holidays. For service information identified in this proposed AD, contact Diamond...

  13. Current Status of the Hot White Dwarf Luminosity Function and non-DA to DA Ratio from SDSS Data

    NASA Astrophysics Data System (ADS)

    Krzesinski, J.; Stachowski, G.

    2017-03-01

    Recent advances in the determination of the hot white dwarf (WD) luminosity function have reached the point where we have good agreement between the observed and theoretical DA and non-DA LFs for WDs from SDSS DR4. The main progress in calculation of the DA LF was obtained when the WD sample was restricted to only carbon-oxygen core WDs. However, one remaining effect which could influence the LF and the non-DA to DA ratio is the difference in visibility of helium- and hydrogen-atmosphere WDs in a magnitude-limited sample. In this work we explore this effect for the SDSS g filter as a function of effective temperature, as well as make an attempt to evaluate data relevant to the WD sample and LFs from more recent data releases of the SDSS.

  14. Effect of 14-kDa and 47-kDa protein molecules of age garlic extract on peritoneal macrophages.

    PubMed

    Daneshmandi, Saeed; Hajimoradi, Monire; Ahmadabad, Hasan Namdar; Hassan, Zuhair Mohammad; Roudbary, Maryam; Ghazanfari, Tooba

    2011-03-01

    Garlic (Allium sativum), traditionally being used as a spice worldwide, has different applications and is claimed to possess beneficial effects in several health ailments such as tumor and atherosclerosis. Garlic is also an immunomodulator and its different components are responsible for different properties. The present work aimed to assess the effect of protein fractions of garlic on peritoneal macrophages. 14-kDa and 47-kDa protein fractions of garlic were purified. Mice peritoneal macrophages were lavaged and cultured in a microtiter plate and exposed to different concentrations of garlic proteins. MTT assay was performed to evaluate the viability of macrophage. The amount of nitric oxide (NO) was detected in culture supernatants of macrophages by Griess reagent and furthermore, the cytotoxicity study of culture supernatants was carried out on WEHI-164 fibrosarcoma cell line as tumor necrosis factor-α bioassay. MTT assay results for both 14-kDa and 47-kDa protein fractions of stimulated macrophages were not significant (P > 0.05). Both 14-kDa and 47-kDa fractions significantly suppressed production of NO from macrophages (P = 0.007 and P = 0.003, respectively). Cytotoxicity of macrophages' supernatant on WEHI-164 fibrosarcoma cells was not affected by garlic protein fractions (P = 0.066 for 14-kDa and P = 0.085 for 47-kDa fractions). according to our finding, 14-kDa and 47-kDa fractions of aged garlic extract are able to suppress NO production from macrophages, which can be used as a biological advantage. These molecules had no cytotoxic effect on macrophages and do not increase tumoricidal property of macrophages.

  15. Beam position monitor system of DA{phi}NE

    SciTech Connect

    Stella, A.; Drago, A.; Ghigo, A.; Marcellini, F.; Milardi, C.; Sannibale, F.; Serio, M.; Vaccarezza, C.

    1998-12-10

    The DA{phi}NE beam position monitor (BPM) system consists of 150 monitors installed all along the machine. Design issues, calibration procedures, experimental results and performance of the system are described. The closed orbit in the main rings is extracted from the BPM signals through narrowband receivers (realized by Bergoz Precision Beam Instrumentation for DA{phi}NE), then acquired and processed by a real-time task based on four independent processors dealing with different machine areas. The data acquisition system is integrated in the DA{phi}NE control system and measures five complete orbits in a second. Implementation criteria, measurements and results are reported.

  16. Circumstellar features in hot DA white dwarfs

    NASA Astrophysics Data System (ADS)

    Bannister, N. P.; Barstow, M. A.; Holberg, J. B.; Bruhweiler, F. C.

    2003-05-01

    We present a phenomenological study of highly ionized, non-photospheric absorption features in high spectral resolution vacuum ultraviolet spectra of 23 hot DA white dwarfs. Prior to this study, four of the survey objects (Feige 24, REJ 0457-281, G191-B2B and REJ 1614-085) were known to possess these features. We find four new objects with multiple components in one or more of the principal resonance lines: REJ 1738+665, Ton 021, REJ 0558-373 and WD 2218+706. A fifth object, REJ 2156-546, also shows some evidence of multiple components, though further observations are required to confirm the detection. We discuss possible origins for these features including ionization of the local interstellar environment, the presence of material inside the gravitational well of the white dwarf, mass loss in a stellar wind and the existence of material in an ancient planetary nebula around the star. We propose ionization of the local interstellar medium as the origin of these features in G191-B2B and REJ 1738+665, and demonstrate the need for higher-resolution spectroscopy of the sample, to detect multiple interstellar medium velocity components and to identify circumstellar features that may lie close to the photospheric velocity.

  17. Phantom surfaces in da Vinci stereopsis.

    PubMed

    Wardle, Susan G; Gillam, Barbara J

    2013-02-08

    In binocular viewing of natural three-dimensional scenes, occlusion relationships between objects at different depths create regions of the background that are visible to only one eye. These monocular regions can support depth perception. There are two viewing conditions in which a monocular region can be on the nasal side of a binocular surface--(a) when a background surface is viewed through an aperture and (b) when a region is camouflaged against the background in one eye's view. We created stimuli with a monocular region using complex textures in which camouflage was not possible, and for which there was no physical aperture. For these stimuli, observers perceived a strong phantom contour in near depth at the edge of the monocular region, with the monocular texture perceived behind at the depth of the binocular surface. Depth-matching with a probe showed that the depth of the phantom occluding surface was as precise as for stimuli with regular binocular disparity. Monocular regions of texture on the opposite (temporal) side of the binocular surface were perceived behind, as predicted by occlusion geometry, and there was no phantom surface. We discuss the implications for models of da Vinci stereopsis and stereoscopic edge processing, and consider the involvement of a form of Panum's limiting case. We conclude that the visual system uses a combination of occlusion geometry and complex matching to precisely locate edges in depth that lack a luminance contour.

  18. Der Telemanipulator daVinci als mechanisches Trackingsystem

    NASA Astrophysics Data System (ADS)

    Käst, Johannes; Neuhaus, Jochen; Nickel, Felix; Kenngott, Hannes; Engel, Markus; Short, Elaine; Reiter, Michael; Meinzer, Hans-Peter; Maier-Hein, Lena

    Der Telemanipulator daVinci (Intuitive Surgical, Sunnyvale, Kalifornien) ist ein M aster-Slave System für roboterassistierte minimalinvasive Chirurgie. Da er über integrierte Gelenksensoren verfügt, kann er unter Verwendung der daVinci-API als mechanisches Trackingsystem verwendet werden. In dieser Arbeit evaluieren wir die Präzision und Genauigkeit eines daVinci mit Hilfe eines Genauigkeitsphantoms mit bekannten Maßen. Der ermittelte Positionierungsfehler liegt in der Größenordnung von 6 mm und ist somit für einen Großteil der medizinischen Fragestellungen zu hoch. Zur Reduktion des Fehlers schlagen wir daher eine Kalibrierung der Gelenksensoren vor.

  19. 40 CFR 60.48Da - Compliance provisions.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... successive boiler operating days is completed within 60 days after achieving the maximum production rate at... determine compliance by using the CEMS specified under § 60.49Da for measuring NOX and oxygen (O2) (or...

  20. 40 CFR 60.48Da - Compliance provisions.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... after achieving the maximum production rate at which the affected facility will be operated, but not... elect to determine compliance by using the CEMS specified under § 60.49Da for measuring NOX and oxygen...

  1. 40 CFR 60.51Da - Reporting requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Generating Units § 60.51Da Reporting requirements. (a) For SO2, NOX, PM, and NOX plus CO emissions, the... emission rates (so) and inlet emission rates (si) as applicable. (3) The lower confidence limit for the...

  2. 40 CFR 60.51Da - Reporting requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Generating Units § 60.51Da Reporting requirements. (a) For SO2, NOX, PM, and NOX plus CO emissions, the... emission rates (so) and inlet emission rates (si) as applicable. (3) The lower confidence limit for the...

  3. Interaction of Arabidopsis TGA3 and WRKY53 transcription factors on Cestrum yellow leaf curling virus (CmYLCV) promoter mediates salicylic acid-dependent gene expression in planta.

    PubMed

    Sarkar, Shayan; Das, Abhimanyu; Khandagale, Prashant; Maiti, Indu B; Chattopadhyay, Sudip; Dey, Nrisingha

    2017-09-14

    This paper highlighted a salicylic acid-inducible Caulimoviral promoter fragment from Cestrum yellow leaf curling virus (CmYLCV). Interaction of Arabidopsis transcription factors TGA3 and WRKY53 on CmYLCV promoter resulted in the enhancement of the promoter activity via NPR1-dependent salicylic acid signaling. Several transcriptional promoters isolated from plant-infecting Caulimoviruses are being presently used worldwide as efficient tools for plant gene expression. The CmYLCV promoter has been isolated from the Cestrum yellow leaf curling virus (Caulimoviruses) and characterized more than 12 years ago; also we have earlier reported a near-constitutive, pathogen-inducible CmYLCV promoter fragment (-329 to +137 from transcription start site; TSS) that enhances stronger (3×) expression than the previously reported fragments; all these fragments are highly efficient in monocot and dicot plants (Sahoo et al. Planta 240: 855-875, 2014). Here, we have shown that the full-length CmYLCV promoter fragment (-729 to +137 from TSS) is salicylic acid (SA) inducible. In this context, we have performed an in-depth study to elucidate the factors responsible for SA-inducibility of the CmYLCV promoter. We found that the as-1 1 and W-box1 elements (located at -649 and -640 from the TSS) of the CmYLCV promoter are required for SA-induced activation by recruiting Arabidopsis TGA3 and WRKY53 transcription factors. Consequently, as a nascent observation, we established the physical interaction between TGA3 and WYKY53; also demonstrated that the N-terminal domain of TGA3 is sufficient for the interaction with the full-length WRKY53. Such interaction synergistically activates the CmYLCV promoter activity in planta. Further, we found that activation of the CmYLCV promoter by SA through TGA3 and WRKY53 interaction depends on NPR1. Finally, the findings presented here provide strong support for the direct regulatory roles of TGA3 and WRKY53 in the SA and NPR1-dependent activation of a

  4. ECTA/DaSy Framework Self-Assessment Comparison Tool

    ERIC Educational Resources Information Center

    Center for IDEA Early Childhood Data Systems (DaSy), 2016

    2016-01-01

    The Self-Assessment Comparison (SAC) Tool is for state Part C and Section 619/Preschool programs to use to assess changes in the implementation of one or more components of the ECTA System Framework and/or subcomponenets of the DaSy Data System Framework. It is a companion to the ECTA/DaSy Framework Self-Assessment. Key features of the SAC are…

  5. Tree branching: Leonardo da Vinci's rule versus biomechanical models.

    PubMed

    Minamino, Ryoko; Tateno, Masaki

    2014-01-01

    This study examined Leonardo da Vinci's rule (i.e., the sum of the cross-sectional area of all tree branches above a branching point at any height is equal to the cross-sectional area of the trunk or the branch immediately below the branching point) using simulations based on two biomechanical models: the uniform stress and elastic similarity models. Model calculations of the daughter/mother ratio (i.e., the ratio of the total cross-sectional area of the daughter branches to the cross-sectional area of the mother branch at the branching point) showed that both biomechanical models agreed with da Vinci's rule when the branching angles of daughter branches and the weights of lateral daughter branches were small; however, the models deviated from da Vinci's rule as the weights and/or the branching angles of lateral daughter branches increased. The calculated values of the two models were largely similar but differed in some ways. Field measurements of Fagus crenata and Abies homolepis also fit this trend, wherein models deviated from da Vinci's rule with increasing relative weights of lateral daughter branches. However, this deviation was small for a branching pattern in nature, where empirical measurements were taken under realistic measurement conditions; thus, da Vinci's rule did not critically contradict the biomechanical models in the case of real branching patterns, though the model calculations described the contradiction between da Vinci's rule and the biomechanical models. The field data for Fagus crenata fit the uniform stress model best, indicating that stress uniformity is the key constraint of branch morphology in Fagus crenata rather than elastic similarity or da Vinci's rule. On the other hand, mechanical constraints are not necessarily significant in the morphology of Abies homolepis branches, depending on the number of daughter branches. Rather, these branches were often in agreement with da Vinci's rule.

  6. ECTA/DaSy Framework Self-Assessment Comparison Tool

    ERIC Educational Resources Information Center

    Center for IDEA Early Childhood Data Systems (DaSy), 2016

    2016-01-01

    The Self-Assessment Comparison (SAC) Tool is for state Part C and Section 619/Preschool programs to use to assess changes in the implementation of one or more components of the ECTA System Framework and/or subcomponenets of the DaSy Data System Framework. It is a companion to the ECTA/DaSy Framework Self-Assessment. Key features of the SAC are…

  7. Migration of ATLAS PanDA to CERN

    NASA Astrophysics Data System (ADS)

    Stewart, Graeme Andrew; Klimentov, Alexei; Koblitz, Birger; Lamanna, Massimo; Maeno, Tadashi; Nevski, Pavel; Nowak, Marcin; Emanuel De Castro Faria Salgado, Pedro; Wenaus, Torre

    2010-04-01

    The ATLAS Production and Distributed Analysis System (PanDA) is a key component of the ATLAS distributed computing infrastructure. All ATLAS production jobs, and a substantial amount of user and group analysis jobs, pass through the PanDA system, which manages their execution on the grid. PanDA also plays a key role in production task definition and the data set replication request system. PanDA has recently been migrated from Brookhaven National Laboratory (BNL) to the European Organization for Nuclear Research (CERN), a process we describe here. We discuss how the new infrastructure for PanDA, which relies heavily on services provided by CERN IT, was introduced in order to make the service as reliable as possible and to allow it to be scaled to ATLAS's increasing need for distributed computing. The migration involved changing the backend database for PanDA from MySQL to Oracle, which impacted upon the database schemas. The process by which the client code was optimised for the new database backend is discussed. We describe the procedure by which the new database infrastructure was tested and commissioned for production use. Operations during the migration had to be planned carefully to minimise disruption to ongoing ATLAS offline computing. All parts of the migration were fully tested before commissioning the new infrastructure and the gradual migration of computing resources to the new system allowed any problems of scaling to be addressed.

  8. DaTo: an atlas of biological databases and tools.

    PubMed

    Li, Qilin; Zhou, Yincong; Jiao, Yingmin; Zhang, Zhao; Bai, Lin; Tong, Li; Yang, Xiong; Sommer, Björn; Hofestädt, Ralf; Chen, Ming

    2016-12-18

    This work presents DaTo, a semi-automatically generated world atlas of biological databases and tools. It extracts raw information from all PubMed articles which contain exact URLs in their abstract section, followed by a manual curation of the abstract and the URL accessibility. DaTo features a user-friendly query interface, providing extensible URL-related annotations, such as the status, the location and the country of the URL. A graphical interaction network browser has also been integrated into the DaTo web interface to facilitate exploration of the relationship between different tools and databases with respect to their ontology-based semantic similarity. Using DaTo, the geographical locations, the health statuses, as well as the journal associations were evaluated with respect to the historical development of bioinformatics tools and databases over the last 20 years. We hope it will inspire the biological community to gain a systematic insight into bioinformatics resources. DaTo is accessible via http://bis.zju.edu.cn/DaTo/.

  9. Mycobacterium tuberculosis 19-kDa lipoprotein promotes neutrophil activation.

    PubMed

    Neufert, C; Pai, R K; Noss, E H; Berger, M; Boom, W H; Harding, C V

    2001-08-01

    Certain microbial substances, e.g., LPS, can activate neutrophils or prime them to enhance their response to other activating agents, e.g., fMLP. We investigated the role of the Mycobacterium tuberculosis (MTB) 19-kDa lipoprotein in activation of human neutrophils. MTB 19-kDa lipoprotein initiated phenotypic changes characteristic of neutrophil activation, including down-regulation of CD62 ligand (L-selectin) and up-regulation of CD35 (CR1) and CD11b/CD18 (CR3, Mac-1). In addition, exposure of neutrophils to MTB 19-kDa lipoprotein enhanced the subsequent oxidative burst in response to fMLP as assessed by oxidation of dihydrorhodamine 123 (determined by flow cytometry). LPS also produced these effects with similar kinetics, but an oligodeoxynucleotide containing a CpG motif failed to induce any priming or activation response. Although the effects of LPS required the presence of serum, neutrophil activation by MTB 19-kDa lipoprotein occurred independently of serum factors, suggesting the involvement of different receptors and signaling mechanisms for LPS and MTB 19-kDa lipoprotein. Thus, MTB 19-kDa lipoprotein serves as a pathogen-associated molecular pattern that promotes neutrophil priming and activation.

  10. The Ubiquitin Receptor DA1 Interacts with the E3 Ubiquitin Ligase DA2 to Regulate Seed and Organ Size in Arabidopsis[C][W

    PubMed Central

    Xia, Tian; Li, Na; Dumenil, Jack; Li, Jie; Kamenski, Andrei; Bevan, Michael W.; Gao, Fan; Li, Yunhai

    2013-01-01

    Seed size in higher plants is determined by the coordinated growth of the embryo, endosperm, and maternal tissue. Several factors that act maternally to regulate seed size have been identified, such as AUXIN RESPONSE FACTOR2, APETALA2, KLUH, and DA1, but the genetic and molecular mechanisms of these factors in seed size control are almost totally unknown. We previously demonstrated that the ubiquitin receptor DA1 acts synergistically with the E3 ubiquitin ligase ENHANCER1 OF DA1 (EOD1)/BIG BROTHER to regulate the final size of seeds in Arabidopsis thaliana. Here, we describe another RING-type protein with E3 ubiquitin ligase activity, encoded by DA2, which regulates seed size by restricting cell proliferation in the maternal integuments of developing seeds. The da2-1 mutant forms large seeds, while overexpression of DA2 decreases seed size of wild-type plants. Overexpression of rice (Oryza sativa) GRAIN WIDTH AND WEIGHT2, a homolog of DA2, restricts seed growth in Arabidopsis. Genetic analyses show that DA2 functions synergistically with DA1 to regulate seed size, but does so independently of EOD1. Further results reveal that DA2 interacts physically with DA1 in vitro and in vivo. Therefore, our findings define the genetic and molecular mechanisms of three ubiquitin-related proteins DA1, DA2, and EOD1 in seed size control and indicate that they are promising targets for crop improvement. PMID:24045020

  11. The ubiquitin receptor DA1 interacts with the E3 ubiquitin ligase DA2 to regulate seed and organ size in Arabidopsis.

    PubMed

    Xia, Tian; Li, Na; Dumenil, Jack; Li, Jie; Kamenski, Andrei; Bevan, Michael W; Gao, Fan; Li, Yunhai

    2013-09-01

    Seed size in higher plants is determined by the coordinated growth of the embryo, endosperm, and maternal tissue. Several factors that act maternally to regulate seed size have been identified, such as auxin response factor2, apetala2, KLUH, and DA1, but the genetic and molecular mechanisms of these factors in seed size control are almost totally unknown. We previously demonstrated that the ubiquitin receptor DA1 acts synergistically with the E3 ubiquitin ligase enhancer1 OF DA1 (EOD1)/big brother to regulate the final size of seeds in Arabidopsis thaliana. Here, we describe another RING-type protein with E3 ubiquitin ligase activity, encoded by DA2, which regulates seed size by restricting cell proliferation in the maternal integuments of developing seeds. The da2-1 mutant forms large seeds, while overexpression of DA2 decreases seed size of wild-type plants. Overexpression of rice (Oryza sativa) grain width and weight2, a homolog of DA2, restricts seed growth in Arabidopsis. Genetic analyses show that DA2 functions synergistically with DA1 to regulate seed size, but does so independently of EOD1. Further results reveal that DA2 interacts physically with DA1 in vitro and in vivo. Therefore, our findings define the genetic and molecular mechanisms of three ubiquitin-related proteins DA1, DA2, and EOD1 in seed size control and indicate that they are promising targets for crop improvement.

  12. Mevalocidin: a novel, phloem mobile phytotoxin from Fusarium DA056446 and Rosellinia DA092917.

    PubMed

    Gerwick, B Clifford; Brewster, William K; Deboer, Gerrit J; Fields, Steve C; Graupner, Paul R; Hahn, Donald R; Pearce, Cedric J; Schmitzer, Paul R; Webster, Jeffery D

    2013-02-01

    A multiyear effort to identify new natural products was built on a hypothesis that both phytotoxins from plant pathogens and antimicrobial compounds might demonstrate herbicidal activity. The discovery of one such compound, mevalocidin, is described in the current report. Mevalocidin was discovered from static cultures of two unrelated fungal isolates designated Rosellinia DA092917 and Fusarium DA056446. The chemical structure was confirmed by independent synthesis. Mevalocidin demonstrated broad spectrum post-emergence activity on grasses and broadleaves and produced a unique set of visual symptoms on treated plants suggesting a novel mode of action. Mevalocidin was rapidly absorbed in a representative grass and broadleaf plant. Translocation occurred from the treated leaf to other plant parts including roots confirming phloem as well as xylem mobility. By 24 hr after application, over 20 % had been redistributed through-out the plant. Mevalocidin is a unique phytotoxin based on its chemistry, with the uncommon attribute of demonstrating both xylem and phloem mobility in grass and broadleaf plants.

  13. Secretion of 10-kDa and 12-kDa thioredoxin species from blood monocytes and transformed leukocytes.

    PubMed

    Sahaf, B; Rosén, A

    2000-01-01

    Thioredoxins (TRX) are ubiquitous, small redox-active proteins with multiple functions, including antioxidant, cytoprotective, and chemoattractant activities. In addition to a 12-kDa intracellular form, extracellular 10-kDa and 12-kDa TRX have been defined. The biological activities of the 10-kDa TRX were previously measured as eosinophil cytotoxicity enhancing activity or B-cell stimulatory activity. Cytotrophoblastic cell lines also release a 10-kDa TRX form. To study the biological role of 10-kDa TRX, we established two highly sensitive enzyme-linked immuno-spot assays (ELISPOT), which detect secreted truncated 10-kDa and full-length 12-kDa TRX at the single cell level. TRX secretion was investigated in several cell lines including the T-helper cell hybridoma MP6, the Jurkat T-cell leukemia, the U-937 myelomonocytic leukemia, and the 3B6, EBV-transformed, lymphoblastoid B-cell line. The highest number of secreting cells was found in 3B6 cultures, median = 34 (quartiles, 27-39) per well (10(5) cells). Peripheral blood monocytes isolated from healthy donors secreted significantly more TRX after stimulation with ionomycin, phorbol myristate acetate (PMA), fMLP, and lipopolysaccharide (LPS), compared to unstimulated cells. Oxidative stress induced by thioloxidant diamide also induced the secretion of both truncated and full-length TRX measured in ELISPOT (p = 0.047 and p = 0.031, respectively). The biological activity of the truncated and full-length forms was tested in a cell migration assay. Truncated TRX was devoid of protein disulfide reductase activity, but retained strong chemoattractant activity for human monocytes, in the same range as full-length TRX, as previously reported (Bertini et al., 1999).

  14. PKC phosphorylates residues in the N-terminal of the DA transporter to regulate amphetamine-induced DA efflux.

    PubMed

    Wang, Qiang; Bubula, Nancy; Brown, Jason; Wang, Yunliang; Kondev, Veronika; Vezina, Paul

    2016-05-27

    The DA transporter (DAT), a phosphoprotein, controls extracellular dopamine (DA) levels in the central nervous system through transport or reverse transport (efflux). Multiple lines of evidence support the claim that PKC significantly contributes to amphetamine-induced DA efflux. Other signaling pathways, involving CaMKII and ERK, have also been shown to regulate DAT mediated efflux. Here we assessed the contribution of putative PKC residues (S4, S7, S13) in the N-terminal of the DAT to amphetamine-induced DA efflux by transfecting DATs containing different serine to alanine (S-A) point mutations into DA pre-loaded HEK-293 cells and incubating these cells in amphetamine (2μM). The effects of a S-A mutation at the non-PKC residue S12 and a threonine to alanine (T-A) mutation at the ERK T53 residue were also assessed for comparison. WT-DATs were used as controls. In an initial experiment, we confirmed that inhibiting PKC with Go6976 (130nM) significantly reduced amphetamine-induced DA efflux. In subsequent experiments, cells transfected with the S4A, S12A, S13A, T53A and S4,7,13A mutants showed a reduction in amphetamine-induced DA efflux similar to that observed with Go6976. Interestingly, cells transfected with the S7A mutant, identified by some as a PKC-PKA residue, showed unperturbed WT-DAT levels of amphetamine-induced DA efflux. These results indicate that phosphorylation by PKC of select residues in the DAT N-terminal can regulate amphetamine-induced efflux. PKC can act either independently or in concert with other kinases such as ERK to produce this effect.

  15. Purification and characterization of 94kDa and 80kDa forms of the muscarinic cholinergic receptor

    SciTech Connect

    Fracek, S.P. Jr.; Venter, J.C.; Kerlavage, A.R.

    1986-05-01

    Two molecular forms of the muscarinic cholinergic receptor have been consistently observed in a variety of species, albeit in variable amounts. Proteins which are specifically labeled by (/sup 3/H)propylbenzilylcholine mustard ((/sup 3/H)PrBCM) were observed at 94kDa and 80kDa upon SDS-PAGE of membrane proteins prepared from brains and hearts of trout, frog, turtle, chicken, rat, and pig. They have developed a purification procedure which yields each of these proteins in a homogeneous form suitable for structural analysis. The four step procedure involves affinity chromatography on 3-(2'-aminobenzhydryloxy)tropane-sepharose, concentration on hydroxylapatite, preparative SDS-PAGE and extraction of individual bands from the gel. Limited tryptic digestion of purified (/sup 3/H)PrBCM-labeled porcine atrial muscarinic receptor yields (/sup 3/H)-labeled fragments of 75, 65, 52, 40, 35, 30, 25, and 20kDa, in close agreement with results of analogous digestions of muscarinic receptor from other species and tissues. Complete tryptic digestion and subsequent mapping by reverse-phase HPLC yields very similar profiles for (/sup 125/I)-labeled 94kDa and 80kDA receptor forms. Most peaks which elute in the hydrophobic region of the profile overlap for the two proteins while the 94kDa protein contains several additional peaks of apparent low hydrophobicity.

  16. An updated model for nitrate uptake modelling in plants. I. Functional component: cross-combination of flow–force interpretation of nitrate uptake isotherms, and environmental and in planta regulation of nitrate influx

    PubMed Central

    Le Deunff, Erwan; Malagoli, Philippe

    2014-01-01

    Background and Aims In spite of major breakthroughs in the last three decades in the identification of root nitrate uptake transporters in plants and the associated regulation of nitrate transport activities, a simplified and operational modelling approach for nitrate uptake is still lacking. This is due mainly to the difficulty in linking the various regulations of nitrate transport that act at different levels of time and on different spatial scales. Methods A cross-combination of a Flow–Force approach applied to nitrate influx isotherms and experimentally determined environmental and in planta regulation is used to model nitrate in oilseed rape, Brassica napus. In contrast to ‘Enzyme–Substrate’ interpretations, a Flow–Force modelling approach considers the root as a single catalytic structure and does not infer hypothetical cellular processes among nitrate transporter activities across cellular layers in the mature roots. In addition, this approach accounts for the driving force on ion transport based on the gradient of electrochemical potential, which is more appropriate from a thermodynamic viewpoint. Key Results and Conclusions Use of a Flow–Force formalism on nitrate influx isotherms leads to the development of a new conceptual mechanistic basis to model more accurately N uptake by a winter oilseed rape crop under field conditions during the whole growth cycle. This forms the functional component of a proposed new structure–function mechanistic model of N uptake. PMID:24638820

  17. 76 FR 12627 - Airworthiness Directives; Diamond Aircraft Industries GmbH Models DA 42, DA 42 NG, and DA 42 M-NG...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-08

    ... Federal Aviation Administration 14 CFR Part 39 RIN 2120-AA64 Airworthiness Directives; Diamond Aircraft... on Diamond aeroplanes, the majority of which were DA 40. In additional, at least 18 doors have been replaced because of damage found on the hinge. Diamond Aircraft Industries conducted analyses...

  18. A cosmologia no ensino da geografia

    NASA Astrophysics Data System (ADS)

    Santos, S. C.; Chiaradia, A. P. M.

    2003-08-01

    O principal objetivo deste trabalho é auxiliar o professor de Geografia em sala de aula no ensino de tópicos relacionados com a Cosmologia. A idéia deste trabalho surgiu quando foi constatado que o professor de Geografia tem dificuldades de ensinar este tópico. Esta constatação foi feita por uma das autoras ao lecionar este tópico no ensino fundamental e em discussões com outros professores de Geografia. Da mesma maneira que ocorria desde os tempos mais antigos, os alunos têm muito interesse em conhecer os fenômenos que ocorrem no Cosmo, porém os livros didáticos de Geografia utilizados em sala de aula não são ricos em informações sobre este assunto. Assim, o professor de Geografia tem poucas informações para discutir este assunto em sala de aula e não dá a devida importância para este tópico. Então, foi desenvolvido um material de apoio para professores de Geografia sobre a origem do Universo, sua evolução e seu possível futuro evolutivo segundo as mais recentes teorias, com base em perguntas feitas pelos alunos de ensino fundamental e as informações trazidas nos livros didáticos Não cabe a este material inovar e tão pouco trazer uma metodologia de ensino de Cosmologia. Neste material o professor de Geografia pode encontrará um banco de informações, que constitui no estabelecimento de conceitos, teorias e hipóteses, sobre a Cosmologia, em linguagem simples e de fácil entendimento. Para desenvolvê-lo, foram feitas pesquisas não exaustivas em livros e revistas científicas, compilação e discussão em forma cronológica das teorias aceitas sobre modelos cosmológicos. Portanto, este material será apresentado neste trabalho.

  19. Overview of ATLAS PanDA Workload Management

    SciTech Connect

    Maeno T.; De K.; Wenaus T.; Nilsson P.; Stewart G. A.; Walker R.; Stradling A.; Caballero J.; Potekhin M.; Smith D.

    2011-01-01

    The Production and Distributed Analysis System (PanDA) plays a key role in the ATLAS distributed computing infrastructure. All ATLAS Monte-Carlo simulation and data reprocessing jobs pass through the PanDA system. We will describe how PanDA manages job execution on the grid using dynamic resource estimation and data replication together with intelligent brokerage in order to meet the scaling and automation requirements of ATLAS distributed computing. PanDA is also the primary ATLAS system for processing user and group analysis jobs, bringing further requirements for quick, flexible adaptation to the rapidly evolving analysis use cases of the early datataking phase, in addition to the high reliability, robustness and usability needed to provide efficient and transparent utilization of the grid for analysis users. We will describe how PanDA meets ATLAS requirements, the evolution of the system in light of operational experience, how the system has performed during the first LHC data-taking phase and plans for the future.

  20. Overview of ATLAS PanDA Workload Management

    NASA Astrophysics Data System (ADS)

    Maeno, T.; De, K.; Wenaus, T.; Nilsson, P.; Stewart, G. A.; Walker, R.; Stradling, A.; Caballero, J.; Potekhin, M.; Smith, D.; ATLAS Collaboration

    2011-12-01

    The Production and Distributed Analysis System (PanDA) plays a key role in the ATLAS distributed computing infrastructure. All ATLAS Monte-Carlo simulation and data reprocessing jobs pass through the PanDA system. We will describe how PanDA manages job execution on the grid using dynamic resource estimation and data replication together with intelligent brokerage in order to meet the scaling and automation requirements of ATLAS distributed computing. PanDA is also the primary ATLAS system for processing user and group analysis jobs, bringing further requirements for quick, flexible adaptation to the rapidly evolving analysis use cases of the early datataking phase, in addition to the high reliability, robustness and usability needed to provide efficient and transparent utilization of the grid for analysis users. We will describe how PanDA meets ATLAS requirements, the evolution of the system in light of operational experience, how the system has performed during the first LHC data-taking phase and plans for the future.

  1. The ATLAS PanDA Monitoring System and its Evolution

    NASA Astrophysics Data System (ADS)

    Klimentov, A.; Nevski, P.; Potekhin, M.; Wenaus, T.

    2011-12-01

    The PanDA (Production and Distributed Analysis) Workload Management System is used for ATLAS distributed production and analysis worldwide. The needs of ATLAS global computing imposed challenging requirements on the design of PanDA in areas such as scalability, robustness, automation, diagnostics, and usability for both production shifters and analysis users. Through a system-wide job database, the PanDA monitor provides a comprehensive and coherent view of the system and job execution, from high level summaries to detailed drill-down job diagnostics. It is (like the rest of PanDA) an Apache-based Python application backed by Oracle. The presentation layer is HTML code generated on the fly in the Python application which is also responsible for managing database queries. However, this approach is lacking in user interface flexibility, simplicity of communication with external systems, and ease of maintenance. A decision was therefore made to migrate the PanDA monitor server to Django Web Application Framework and apply JSON/AJAX technology in the browser front end. This allows us to greatly reduce the amount of application code, separate data preparation from presentation, leverage open source for tools such as authentication and authorization mechanisms, and provide a richer and more dynamic user experience. We describe our approach, design and initial experience with the migration process.

  2. Vínculos sobre um modelo de quartessência de Chaplygin usando observações do satélite chandra da fração de massa de gás em aglomerados de galáxias

    NASA Astrophysics Data System (ADS)

    de Souza, R. S.

    2003-08-01

    Observações de Supernovas do tipo Ia mostram que a expansão do Universo está acelerando. Segundo as equações de Einstein uma componente com pressão negativa (energia escura) é necessária para explicar a aceleração cósmica. Além da energia escura é usualmente admitido que no Universo há também uma matéria exótica com pressão zero, que é chamada de matéria escura. Essa componente possui um papel fundamental na formação de estruturas no Universo. Recentemente tem se explorado a possibilidade de que matéria e energia escura poderiam ser unificadas através de uma única componente, que tem sido denominada de quartessência. Um exemplo de fluido com essas características é o Gás de Chaplygin Generalizado, que possui uma equação de estado da forma p = -A/ra. Inicialmente consideramos o caso especial a = 1 (gás de Chaplygin) e vinculamos parâmetros do modelo utilizando observações em raios-X do satélite Chandra da fração de massa de gás em aglomerados de galáxias. Uma comparação dos vínculos obtidos com esse teste com outros testes, tais como supernovas e idade do Universo, mostra que esse teste é bastante restritivo. Exibiremos ainda resultados para o caso em que a curvatura é nula e o parâmetro a está compreendido no intervalo -1 < a 1.

  3. Geomagnetic observations on tristan da cunha, south atlantic ocean

    USGS Publications Warehouse

    Matzka, J.; Olsen, N.; Maule, C.F.; Pedersen, L.W.; Berarducci, A.M.; Macmillan, S.

    2009-01-01

    Few geomagnetic ground observations exist of the Earth's strongest core field anomaly, the South Atlantic Anomaly (SAA). The geomagnetic repeat station on the island Tristan da Cunha, located half-way between South Africa and South America at 37?? 05' S, 12?? 18' W, is therefore of crucial importance. We have conducted several sets of repeat station measurements during magnetically quiet conditions (Kp 2o or less) in 2004. The procedures are described and the results are compared to those from earlier campaigns and to the predictions of various global field models. Features of the local crustal bias field and the solar quiet daily variation are discussed. We also evaluate the benefit of continuous magnetic field recordings from Tristan da Cunha, and argue that such a data set is a very valuable addition to geomagnetic satellite data. Recently, funds were set up to establish and operate a magnetometer station on Tristan da Cunha during the Swarm magnetic satellite mission (2011-2014).

  4. 1991 DA: An asteroid in a bizarre orbit

    NASA Technical Reports Server (NTRS)

    Steel, Duncan; Mcnaught, Robert H.; Asher, David

    1992-01-01

    Asteroidal object 1991 DA has an orbit of high inclination, crossing the planets from Mars to Uranus. This is unique for an asteroid, but not unusual for a comet of the Halley-type: it therefore seems likely that 1991 DA is an extinct or dormant comet. Previous CCD imaging has shown no indication of a coma; spectroscopic observations of 1991 DA which lack any evidence of strong comet-like emissions are reported. Numerical integrations of the orbit of this object were performed which show that is has been remarkably stable for the past approximately 20,000 yr, but chaotic before that. This may allow a new estimate to be made of the physical lifetimes of comets.

  5. Situação da Mulher na Astronomia Brasileira

    NASA Astrophysics Data System (ADS)

    Silva, Adriana V. R.

    2007-07-01

    O conteúdo desse texto surgiu de uma apresentação de mesmo título que fiz na XXXI Reunião Anual da Sociedade Astronômica Brasileira (SAB) em 2005. Esse tema foi inspirado originalmente pela minha participação no "2nd UIPAP International Conference on Women in Physics" realizado entre 23 e 25 de maio de 2005 no Rio de Janeiro. Essa é uma conferência internacional que acontece de três em três anos, sendo que a primeira ocorreu em 2002 na cidade de Paris, França. Participei dessa conferência como membro da delegação da Sociedade Brasileira de Física e um dos trabalhos que apresentei versava sobre a situação das mulheres na Astronomia brasileira, cujos resultados principais discorro a seguir. A situação das astrônomas, baseada nos dados dos sócios da SAB coletados no final de 2004, é comparada com a das físicas brasileiras e também com as nossas colegas americanas. Os dados identificam ainda uma maior evasão da carreira por parte das mulheres do que os homens. Alguns dos possíveis motivos da evasão são discutidos, como o desejo de constituir família e/ou isolamento. Resultados um tanto preocupantes com relação à distribuição de bolsas de produtividade do CNPq também são apresentados. As principais discussões e estratégias recomendadas nesse congresso são mencionadas de forma resumida ao final.

  6. The future of PanDA in ATLAS distributed computing

    NASA Astrophysics Data System (ADS)

    De, K.; Klimentov, A.; Maeno, T.; Nilsson, P.; Oleynik, D.; Panitkin, S.; Petrosyan, A.; Schovancova, J.; Vaniachine, A.; Wenaus, T.

    2015-12-01

    Experiments at the Large Hadron Collider (LHC) face unprecedented computing challenges. Heterogeneous resources are distributed worldwide at hundreds of sites, thousands of physicists analyse the data remotely, the volume of processed data is beyond the exabyte scale, while data processing requires more than a few billion hours of computing usage per year. The PanDA (Production and Distributed Analysis) system was developed to meet the scale and complexity of LHC distributed computing for the ATLAS experiment. In the process, the old batch job paradigm of locally managed computing in HEP was discarded in favour of a far more automated, flexible and scalable model. The success of PanDA in ATLAS is leading to widespread adoption and testing by other experiments. PanDA is the first exascale workload management system in HEP, already operating at more than a million computing jobs per day, and processing over an exabyte of data in 2013. There are many new challenges that PanDA will face in the near future, in addition to new challenges of scale, heterogeneity and increasing user base. PanDA will need to handle rapidly changing computing infrastructure, will require factorization of code for easier deployment, will need to incorporate additional information sources including network metrics in decision making, be able to control network circuits, handle dynamically sized workload processing, provide improved visualization, and face many other challenges. In this talk we will focus on the new features, planned or recently implemented, that are relevant to the next decade of distributed computing workload management using PanDA.

  7. The role of transparency in da Vinci stereopsis.

    PubMed

    Zannoli, Marina; Mamassian, Pascal

    2011-10-15

    The majority of natural scenes contains zones that are visible to one eye only. Past studies have shown that these monocular regions can be seen at a precise depth even though there are no binocular disparities that uniquely constrain their locations in depth. In the so-called da Vinci stereopsis configuration, the monocular region is a vertical line placed next to a binocular rectangular occluder. The opacity of the occluder has been mentioned to be a necessary condition to obtain da Vinci stereopsis. However, this opacity constraint has never been empirically tested. In the present study, we tested whether da Vinci stereopsis and perceptual transparency can interact using a classical da Vinci configuration in which the opacity of the occluder varied. We used two different monocular objects: a line and a disk. We found no effect of the opacity of the occluder on the perceived depth of the monocular object. A careful analysis of the distribution of perceived depth revealed that the monocular object was perceived at a depth that increased with the distance between the object and the occluder. The analysis of the skewness of the distributions was not consistent with a double fusion explanation, favoring an implication of occlusion geometry in da Vinci stereopsis. A simple model that includes the geometry of the scene could account for the results. In summary, the mechanism responsible to locate monocular regions in depth is not sensitive to the material properties of objects, suggesting that da Vinci stereopsis is solved at relatively early stages of disparity processing. Copyright © 2011 Elsevier Ltd. All rights reserved.

  8. The DaCHS Multi-protocol VO Server

    NASA Astrophysics Data System (ADS)

    Demleitner, M.

    2014-05-01

    GAVO's Data Center Helper Suite (DaCHS) is a suite of tools for publishing data to the Virtual Observatory. It implements all major VO protocols (SCS, SIAP, SSAP, TAP, OAI-PMH). The integrated management and ingestion component allow defining metadata, structure, and services once and re-use the definition throughout the publication cycle from initial metadata aquisition to registry record generation. It has been driving GAVO's data center since 2008 and is now deployed in multiple locations around the globe. This poster briefly describes the design of the system as well as a bird's-eye view of data publishing with DaCHS.

  9. Using DA White Dwarfs to Calibrate Synthetic Photometry

    NASA Astrophysics Data System (ADS)

    Holberg, J. B.

    2007-04-01

    Four widely used photometric systems, namely the Johnson-Kron-Cousins UBVRI, the Strömgren uvby, the 2MASS JHKs and the Sloan Digital Sky Survey ugriz systems have been directly compared with the HST absolute photometric scale of Bohlin & Gilliland (2004). These comparisons are subsequently used to construct a large grid of accurate synthetic magnitudes for DA white dwarfs. This grid is, in turn, critically evaluated with respect to the observed photometry from substantial samples of actual white dwarfs. The advantages of DA white dwarfs as photometric stars are emphasized, and the prospects for extending the use of these stars into the near infrared are highlighted.

  10. In Planta Processing and Glycosylation of a Nematode CLAVATA3/ENDOSPERM SURROUNDING REGION-Like Effector and Its Interaction with a Host CLAVATA2-Like Receptor to Promote Parasitism1[OPEN

    PubMed Central

    Chen, Shiyan; Lang, Ping; Chronis, Demosthenis; Zhang, Sheng; De Jong, Walter S.; Mitchum, Melissa G.

    2015-01-01

    Like other biotrophic plant pathogens, plant-parasitic nematodes secrete effector proteins into host cells to facilitate infection. Effector proteins that mimic plant CLAVATA3/ENDOSPERM SURROUNDING REGION-related (CLE) proteins have been identified in several cyst nematodes, including the potato cyst nematode (PCN); however, the mechanistic details of this cross-kingdom mimicry are poorly understood. Plant CLEs are posttranslationally modified and proteolytically processed to function as bioactive ligands critical to various aspects of plant development. Using ectopic expression coupled with nanoliquid chromatography-tandem mass spectrometry analysis, we show that the in planta mature form of proGrCLE1, a multidomain CLE effector secreted by PCN during infection, is a 12-amino acid arabinosylated glycopeptide (named GrCLE1-1Hyp4,7g) with striking structural similarity to mature plant CLE peptides. This glycopeptide is more resistant to hydrolytic degradation and binds with higher affinity to a CLAVATA2-like receptor (StCLV2) from potato (Solanum tuberosum) than its nonglycosylated forms. We further show that StCLV2 is highly up-regulated at nematode infection sites and that transgenic potatoes with reduced StCLV2 expression are less susceptible to PCN infection, indicating that interference of the CLV2-mediated signaling pathway confers nematode resistance in crop plants. These results strongly suggest that phytonematodes have evolved to utilize host cellular posttranslational modification and processing machinery for the activation of CLE effectors following secretion into plant cells and highlight the significance of arabinosylation in regulating nematode CLE effector activity. Our finding also provides evidence that multidomain CLEs are modified and processed similarly to single-domain CLEs, adding new insight into CLE maturation in plants. PMID:25416475

  11. Origin and distribution of Calvin cycle fructose and sedoheptulose bisphosphatases in plantae and complex algae: a single secondary origin of complex red plastids and subsequent propagation via tertiary endosymbioses.

    PubMed

    Teich, René; Zauner, Stefan; Baurain, Denis; Brinkmann, Henner; Petersen, Jörn

    2007-07-01

    Sedoheptulose-1,7-bisphosphatase (SBPase) and fructose-1,6-bisphosphatase (FBPase) are essential nuclear-encoded enzymes involved in land plant Calvin cycle and gluconeogenesis. In this study, we cloned seven SBP and seven FBP cDNAs/genes and established sequences from all lineages of photosynthetic eukaryotes, in order to investigate their origin and evolution. Our data are best explained by a single recruitment of plastid-targeted SBP in Plantae after primary endosymbiosis and a further distribution to algae with complex plastids. While SBP is universally found in photosynthetic lineages, its presence in apicomplexa, ciliates, trypanosomes, and ascomycetes is surprising given that no metabolic function beyond the one in the plastid Calvin cycle is described so far. Sequences of haptophytes, cryptophytes, diatoms, and peridinin-containing dinoflagellates (complex red lineage) strongly group together in the SBP tree and the same assemblage is recovered for plastid-targeted FBP sequences, although this is less supported. Both SBP and plastid-targeted FBP are most likely of red algal origin. Including phosphoribulokinase, fructose bisphosphate aldolase, and glyceraldehyde-3-phosphate dehydrogenase, a total of five independent plastid-related nuclear-encoded markers support a common origin of all complex rhodoplasts via a single secondary endosymbiosis event. However, plastid phylogenies are incongruent with those of the host cell, as illustrated by the cytosolic FBP isoenzyme. These results are discussed in the context of Cavalier-Smith's far-reaching chromalveolate hypothesis. In our opinion, a more plausible evolutionary scenario would be the establishment of a unique secondary rhodoplast and its subsequent spread via tertiary endosymbioses.

  12. Training and Health. Leonardo da Vinci Series: Good Practices.

    ERIC Educational Resources Information Center

    Commission of the European Communities, Brussels (Belgium). Directorate-General for Education and Culture.

    This document profiles programs in the fields of health and medicine that are offered through the European Commission's Leonardo da Vinci program. The following programs are profiled: (1) CYTOTRAIN (a transnational vocational training program in cervical cancer screening); (2) Apollo (a program of open and distance learning for paramedical…

  13. 40 CFR 60.49Da - Emission monitoring.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... (except a wet scrubber) is used for reducing PM, SO2, or carbon monoxide (CO) emissions; (B) Only natural..., calibrate, maintain, and operate a CEMS, and record the output of the system, for measuring the O2 or carbon... for which the data capture requirement of § 60.49Da(p)(4)(i) was not met. (iv) Notwithstanding...

  14. Leonardo da Vinci's foot: historical evidence of concept.

    PubMed

    Jastifer, James R; Toledo-Pereyra, Luis H

    2012-10-01

    Leonardo da Vinci (1452-1519), world-renowned Italian renaissance master, is known for his contributions to, and broad interests in science and art. The objective of this work is to demonstrate the extent of his science by applying the use of his concepts to current models of foot and ankle mechanics. The art and science of Leonardo Da Vinci were extensively analyzed by reviewing his original drawings and hand written notebooks as well as their English translation. Current medical journals including the topics of foot, ankle, and biomechanics were reviewed for modern evidence and application of his concepts. The library of Michigan State University and the electronic library of the Royal Library at Windsor Castle were extensively utilized. From the depths of Santa Maria Nuova Hospital in Florence and Santo Spirito Hospital in Rome, through his commentary and anatomical drawings of around 30 cadaver dissections he performed, Leonardo da Vinci expressed his concept of foot and ankle anatomy and mechanics. He laid forth concepts, which vary little from current theories including those of proportion, statics and joint stability, sesamoid biomechanics, and structural support of the foot. Leonardo da Vinci, by combining an interest in anatomy and a gift of genius and artistic ability laid a foundation of foot and ankle anatomy and mechanics that have been applied in modern clinical sciences. Leonardo in this way made important contributions to the practice of foot and ankle orthopedics.

  15. The Potential da Vinci in All of Us

    ERIC Educational Resources Information Center

    Petto, Sarah; Petto, Andrew

    2009-01-01

    The study of the human form is fundamental to both science and art curricula. For vertebrates, perhaps no feature is more important than the skeleton to determine observable form and function. As Leonard da Vinci's famous Proportions of the Human Figure (Virtruvian Man) illustrates, the size, shape, and proportions of the human body are defined by…

  16. The DaVinci Project: Multimedia in Art and Chemistry.

    ERIC Educational Resources Information Center

    Simonson, Michael; Schlosser, Charles

    1998-01-01

    Provides an overview of the DaVinci Project, a collaboration of students, teachers, and researchers in chemistry and art to develop multimedia materials for grades 3-12 visualizing basic concepts in chemistry and visual art. Topics addressed include standards in art and science; the conceptual framework for the project; and project goals,…

  17. 40 CFR 60.47Da - Commercial demonstration permit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 6 2011-07-01 2011-07-01 false Commercial demonstration permit. 60... Steam Generating Units for Which Construction is Commenced After September 18, 1978 § 60.47Da Commercial... technology may apply to the Administrator for a commercial demonstration permit. The Administrator will...

  18. Studying and Working Abroad. Leonardo da Vinci Series: Good Practices.

    ERIC Educational Resources Information Center

    Commission of the European Communities, Brussels (Belgium). Directorate-General for Education and Culture.

    This document profiles recent successful examples of students studying and working abroad as part of the European Commission's Leonardo da Vinci program, which is designed to give students across the European Union the opportunity to experience vocational training in a foreign country. The following examples are presented: (1) 3 Finnish students…

  19. The Potential da Vinci in All of Us

    ERIC Educational Resources Information Center

    Petto, Sarah; Petto, Andrew

    2009-01-01

    The study of the human form is fundamental to both science and art curricula. For vertebrates, perhaps no feature is more important than the skeleton to determine observable form and function. As Leonard da Vinci's famous Proportions of the Human Figure (Virtruvian Man) illustrates, the size, shape, and proportions of the human body are defined by…

  20. The DaVinci Project: Multimedia in Art and Chemistry.

    ERIC Educational Resources Information Center

    Simonson, Michael; Schlosser, Charles

    1998-01-01

    Provides an overview of the DaVinci Project, a collaboration of students, teachers, and researchers in chemistry and art to develop multimedia materials for grades 3-12 visualizing basic concepts in chemistry and visual art. Topics addressed include standards in art and science; the conceptual framework for the project; and project goals,…

  1. Da Que Hablar (Something To Talk About), 1991-1993.

    ERIC Educational Resources Information Center

    Da Que Hablar, 1993

    1993-01-01

    This document consists of all 14 issues of a bimonthly serial, from its inception in May 1991 through November 1993. "Da Que Hablar" provides numerous authentic materials from magazines and newspapers to stimulate discussion in Spanish in the foreign language classroom. The articles cover topics such as current events, cultural issues,…

  2. Linear theory radial and nonradial pulsations of DA dwarf stars

    SciTech Connect

    Starrfield, S.; Cox, A.N.; Hodson, S.; Pesnell, W.D.

    1982-07-28

    The Los Alamos stellar envelope and radial linear non-adiabatic computer code, along with a new Los Alamos non-radial code are used to investigate the total hydrogen mass necessary to produce the non-radial instability of DA dwarfs. (GHT)

  3. Women and Technical Professions. Leonardo da Vinci Series: Good Practices.

    ERIC Educational Resources Information Center

    Commission of the European Communities, Brussels (Belgium). Directorate-General for Education and Culture.

    This document profiles programs for women in technical professions that are offered through the European Commission's Leonardo da Vinci program. The following programs are profiled: (1) Artemis and Diana (vocational guidance programs to help direct girls toward technology-related careers); (2) CEEWIT (an Internet-based information and…

  4. 40 CFR 60.45Da - Standard for mercury (Hg).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Standard for mercury (Hg). 60.45Da... for mercury (Hg). (a) For each coal-fired electric utility steam generating unit other than an IGCC... gases that contain mercury (Hg) emissions in excess of each Hg emissions limit in paragraphs...

  5. Modelagem do vento e da fotosfera de AG Carinae

    NASA Astrophysics Data System (ADS)

    Groh, J. H.; Damineli, A.

    2003-08-01

    A trajetória evolutiva das estrelas de alta massa depende fortemente de suas taxas de perda de massa. Apesar do rápido progresso no estudo destas estrelas, a taxa de perda de massa e outros parâmetros físicos básicos, como a temperatura superficial e a velocidade terminal do vento ainda não estão bem determinados. Isto ocorre devido à presença de ventos irregulares, rápidos e fortes ao redor destas estrelas, tornando a interpretação dos seus espectros uma tarefa difícil. Assim, a modelagem do vento e da fotosfera dessas estrelas está sendo cada vez mais usada para obter tais parâmetros a partir dos espectros. O aumento da taxa de perda de massa durante a fase LBV (Variáveis Luminosas Azuis), comparado com outros tipos de estrelas, tem sido atribuído a instabilidades do tipo S Doradus. Dispomos de uma base de dados espectroscópicos cobrindo 22 anos de observações de AG Carinae, incluindo um ciclo S Doradus completo, com espectros CCD em alta resolução na faixa óptica e infravermelha. Utilizamos o programa desenvolvido por Schmutz (1997) para uma análise preliminar desse ciclo, obtendo a taxa de perda de massa a partir da linha do Ha. Não existe uma correlação clara da taxa de perda de massa com mudanças da temperatura efetiva, do raio da estrela e do fluxo na banda V. A estrela atingiu seu mínimo fotométrico (raio mínimo) em 1990 e o máximo fotométrico (raio máximo) em 1995, enquanto que o fluxo máximo da linha do Ha ocorreu em 1996. Além disso a taxa de perda de massa não segue esse ciclo, contrariamente às idéias correntes. Para fazer um modelo mais realista estamos usando o programa CMFGEN (Hillier & Miller), que trata a fotosfera e o vento estelar de forma consistente, considerando a radiação fora do equilíbrio termodinâmico (NLTE) e com blanketting total de linhas. Simulamos o espectro de AG Carinae em duas épocas extremas do ciclo S Dor para testar os resultados obtidos com o modelo mais simplificado.

  6. 49 CFR 178.58 - Specification 4DA welded steel cylinders for aircraft use.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 3 2014-10-01 2014-10-01 false Specification 4DA welded steel cylinders for...) SPECIFICATIONS FOR PACKAGINGS Specifications for Cylinders § 178.58 Specification 4DA welded steel cylinders for aircraft use. (a) Type, size, and service pressure. A DOT 4DA is a welded steel sphere (two...

  7. 49 CFR 178.58 - Specification 4DA welded steel cylinders for aircraft use.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 3 2012-10-01 2012-10-01 false Specification 4DA welded steel cylinders for...) SPECIFICATIONS FOR PACKAGINGS Specifications for Cylinders § 178.58 Specification 4DA welded steel cylinders for aircraft use. (a) Type, size, and service pressure. A DOT 4DA is a welded steel sphere (two...

  8. 49 CFR 178.58 - Specification 4DA welded steel cylinders for aircraft use.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 3 2013-10-01 2013-10-01 false Specification 4DA welded steel cylinders for...) SPECIFICATIONS FOR PACKAGINGS Specifications for Cylinders § 178.58 Specification 4DA welded steel cylinders for aircraft use. (a) Type, size, and service pressure. A DOT 4DA is a welded steel sphere (two...

  9. 49 CFR 178.58 - Specification 4DA welded steel cylinders for aircraft use.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 3 2011-10-01 2011-10-01 false Specification 4DA welded steel cylinders for...) SPECIFICATIONS FOR PACKAGINGS Specifications for Cylinders § 178.58 Specification 4DA welded steel cylinders for aircraft use. (a) Type, size, and service pressure. A DOT 4DA is a welded steel sphere (two...

  10. Adapter for contact force sensing of the da Vinci robot.

    PubMed

    Shimachi, Shigeyuki; Hirunyanitiwatna, Surakij; Fujiwara, Yasunori; Hashimoto, Akira; Hakozaki, Yoshinori

    2008-06-01

    At present, the da Vinci surgical robot system does not provide haptic feedback. One of the authors has proposed a contact-force sensing method called the 'overcoat method', in which the instrument/driver is supported by force sensors. In the da Vinci robot, the instrument jaws are powered by a wire-pulley mechanism; thus, in order to apply the overcoat method to the da Vinci system, we must transfer the power through a frame that is supported by force sensors. The authors have attempted to add a force-sensor function to the Sterile Adapter of the da Vinci system. In developing a sensorized adapter, a new configuration of force sensors and a new axial-force-free (AFF) joint have been devised in order to obtain an independent 'axial force effect' from the drive torque fed from the da Vinci robot arm. The force-sensing errors of the present system have been measured to have a maximum value of approximately 0.2 N while driving the jaws, and a maximum value of approximately 0.2 N when the robot arm is inclined with some excitation. Some impact reference forces applied on to the ends of the jaws agree with the outputs of the sensorized adapter to within <0.05 N. It is shown that the new adapter can be sterilized. One apprehension is that the total weight of the new adapter-approximately 1.2 kg-might unbalance the robot arm. In the case of the new adapter, the centre-line of the instrument shaft is shifted externally through approximately 3.5 mm from its original position. However, a new cannula for the da Vinci robot might solve this problem. The new configuration of force sensors and the new AFF joint work well in their basic functions. The total force-sensing error is estimated as approximately 0.5 N. One of the main reasons for the error appears to be the deformation of the adapter frame. (c) 2008 John Wiley & Sons, Ltd.

  11. JCoDA: a tool for detecting evolutionary selection

    PubMed Central

    2010-01-01

    Background The incorporation of annotated sequence information from multiple related species in commonly used databases (Ensembl, Flybase, Saccharomyces Genome Database, Wormbase, etc.) has increased dramatically over the last few years. This influx of information has provided a considerable amount of raw material for evaluation of evolutionary relationships. To aid in the process, we have developed JCoDA (Java Codon Delimited Alignment) as a simple-to-use visualization tool for the detection of site specific and regional positive/negative evolutionary selection amongst homologous coding sequences. Results JCoDA accepts user-inputted unaligned or pre-aligned coding sequences, performs a codon-delimited alignment using ClustalW, and determines the dN/dS calculations using PAML (Phylogenetic Analysis Using Maximum Likelihood, yn00 and codeml) in order to identify regions and sites under evolutionary selection. The JCoDA package includes a graphical interface for Phylip (Phylogeny Inference Package) to generate phylogenetic trees, manages formatting of all required file types, and streamlines passage of information between underlying programs. The raw data are output to user configurable graphs with sliding window options for straightforward visualization of pairwise or gene family comparisons. Additionally, codon-delimited alignments are output in a variety of common formats and all dN/dS calculations can be output in comma-separated value (CSV) format for downstream analysis. To illustrate the types of analyses that are facilitated by JCoDA, we have taken advantage of the well studied sex determination pathway in nematodes as well as the extensive sequence information available to identify genes under positive selection, examples of regional positive selection, and differences in selection based on the role of genes in the sex determination pathway. Conclusions JCoDA is a configurable, open source, user-friendly visualization tool for performing evolutionary

  12. Produccion de plantas grandes usando minicontenedores

    Treesearch

    R. Kasten Dumroese; Thomas D. Landis

    2012-01-01

    En América del Norte hay cada vez más interés por la producción híbrida o mixta. La misma consiste en cultivar plantines en contenedores de pequeño volumen y luego trasplantarlos; el trasplante se puede realizar a canteros en el suelo como en la producción a raíz desnuda o bien a contenedores más grandes. Originalmente se llamaban plantines "plug+", "...

  13. PanDaTox: A tool for accelerated metabolic engineering

    SciTech Connect

    Amitai, Gil; Sorek, Rotem

    2012-07-18

    Metabolic engineering is often facilitated by cloning of genes encoding enzymes from various heterologous organisms into E. coli. Such engineering efforts are frequently hampered by foreign genes that are toxic to the E. coli host. We have developed PanDaTox (www.weizmann.ac.il/pandatox), a web-based resource that provides experimental toxicity information for more than 1.5 million genes from hundreds of different microbial genomes. The toxicity predictions, which were extensively experimentally verified, are based on serial cloning of genes into E. coli as part of the Sanger whole genome shotgun sequencing process. PanDaTox can accelerate metabolic engineering projects by allowing researchers to exclude toxic genes from the engineering plan and verify the clonability of selected genes before the actual metabolic engineering experiments are conducted.

  14. [Giovanbattista da Monte (Montanus): father of modern clinical medicine].

    PubMed

    Franceschetti, Diego; Agazia, Bruno; Zanchin, Giorgio

    2005-01-01

    The figure of Giovanbattista da Monte (1489-1551) is associated with the introduction of clinical teaching at the patient's beside, in 1543, at the San Francesco Hospital of Padua. In the XVI century, teaching was still based on the explanation and comment of the ancient authors and the educational programme was founded on theoretical aspects. The "practical" approach consisted of the treatment "ex cathedra" of diseases according to the various parts of the body, without observing the course of the pathological events with a direct confirmation at the patient's beside.To his merit, Da Monte established the practise of training students to gather the case history, to carry out an objective examination, and to closely examine disease phenomena with lessons at the bedside of the patient. Practical clinical training was thus introduced as the crucial moment in the formation of the physician.

  15. TA-DA: A TOOL FOR ASTROPHYSICAL DATA ANALYSIS

    SciTech Connect

    Da Rio, Nicola; Robberto, Massimo

    2012-12-01

    We present the Tool for Astrophysical Data Analysis (TA-DA), a new software aimed to greatly simplify and improve the analysis of stellar photometric data in comparison with theoretical models, and allow the derivation of stellar parameters from multi-band photometry. Its flexibility allows one to address a number of such problems: from the interpolation of stellar models, or sets of stellar physical parameters in general, to the computation of synthetic photometry in arbitrary filters or units; from the analysis of observed color-magnitude diagrams to a Bayesian derivation of stellar parameters (and extinction) based on multi-band data. TA-DA is available as a pre-compiled Interactive Data Language widget-based application; its graphical user interface makes it considerably user-friendly. In this paper, we describe the software and its functionalities.

  16. A novel 29-kDa chicken heat shock protein.

    PubMed

    Einat, M F; Haberfeld, A; Shamay, A; Horev, G; Hurwitz, S; Yahav, S

    1996-12-01

    The family of small heat shock proteins is the more variable among the highly conserved superfamily of heat shock proteins (HSP). Using a metabolic labeling procedure with tissue explants, we have detected in chickens a new member of the small HSP family with an apparent molecular weight of 29-kDa. This protein was induced in broiler chickens' heart muscle and lungs following an in vivo heat stress. The 29-kDa band appears after 3 h of heat stress, much later than the induction of HSP 90, HSP 70, and HSP 27. The late onset of induction suggests that HSP 29 plays a more specific role of a "second stage defense protein".

  17. DaVinci's Mona Lisa entering the next dimension.

    PubMed

    Carbon, Claus-Christian; Hesslinger, Vera M

    2013-01-01

    For several of Leonardo da Vinci's paintings, such as The Virgin and Child with St Anne or the Mona Lisa, there exist copies produced by his own studio. In case of the Mona Lisa, a quite exceptional, rediscovered studio copy was presented to the public in 2012 by the Prado Museum in Madrid. Not only does it mirror its famous counterpart superficially; it also features the very same corrections to the lower layers, which indicates that da Vinci and the 'copyist' must have elaborated their panels simultaneously. On the basis of subjective (thirty-two participants estimated painter-model constellations) as well as objective data (analysis of trajectories between landmarks of both paintings), we revealed that both versions differ slightly in perspective. We reconstructed the original studio setting and found evidence that the disparity between both paintings mimics human binocular disparity. This points to the possibility that the two Giocondas together might represent the first stereoscopic image in world history.

  18. Calibration of Synthetic Photometry Using DA White Dwarfs

    NASA Astrophysics Data System (ADS)

    Holberg, J. B.; Bergeron, P.

    2005-12-01

    We have calibrated four major ground-based photometric systems with respect to the Hubble Space Telescope absolute flux scale, which is defined by Vega and four fundamental DA white dwarfs. These photometric systems include the Johnson-Kron-Cousins UBVRI, the Stromgren uvby filters, the 2MASS JHKs and the Sloan Digital Sky Survey ugriz filters. Synthetic magnitudes are calculated from model white dwarf spectra folded through the published filter response functions, these magnitudes in turn are absolutely calibrated with respect to the HST flux scale. Effective zero magnitude fluxes and zero point offsets of each system are determined. In order to verify the external observational consistency as well as to demonstrate the applicability of these definitions, the synthetic magnitudes are compared with the respective observed magnitudes of larger sets of DA white dwarfs that have well determined effective temperatures and surface gravities and which span a wide range in both of these parameters.

  19. Da Vinci Xi Robot-Assisted Penetrating Keratoplasty.

    PubMed

    Chammas, Jimmy; Sauer, Arnaud; Pizzuto, Joëlle; Pouthier, Fabienne; Gaucher, David; Marescaux, Jacques; Mutter, Didier; Bourcier, Tristan

    2017-06-01

    This study aims (1) to investigate the feasibility of robot-assisted penetrating keratoplasty (PK) using the new Da Vinci Xi Surgical System and (2) to report what we believe to be the first use of this system in experimental eye surgery. Robot-assisted PK procedures were performed on human corneal transplants using the Da Vinci Xi Surgical System. After an 8-mm corneal trephination, four interrupted sutures and one 10.0 monofilament running suture were made. For each procedure, duration and successful completion of the surgery as well as any unexpected events were assessed. The depth of the corneal sutures was checked postoperatively using spectral-domain optical coherence tomography (SD-OCT). Robot-assisted PK was successfully performed on 12 corneas. The Da Vinci Xi Surgical System provided the necessary dexterity to perform the different steps of surgery. The mean duration of the procedures was 43.4 ± 8.9 minutes (range: 28.5-61.1 minutes). There were no unexpected intraoperative events. SD-OCT confirmed that the sutures were placed at the appropriate depth. We confirm the feasibility of robot-assisted PK with the new Da Vinci Surgical System and report the first use of the Xi model in experimental eye surgery. Operative time of robot-assisted PK surgery is now close to that of conventional manual surgery due to both improvement of the optical system and the presence of microsurgical instruments. Experimentations will allow the advantages of robot-assisted microsurgery to be identified while underlining the improvements and innovations necessary for clinical use.

  20. Improving Security in the ATLAS PanDA System

    NASA Astrophysics Data System (ADS)

    Caballero, J.; Maeno, T.; Nilsson, P.; Stewart, G.; Potekhin, M.; Wenaus, T.

    2011-12-01

    The security challenges faced by users of the grid are considerably different to those faced in previous environments. The adoption of pilot jobs systems by LHC experiments has mitigated many of the problems associated with the inhomogeneities found on the grid and has greatly improved job reliability; however, pilot jobs systems themselves must then address many security issues, including the execution of multiple users' code under a common 'grid' identity. In this paper we describe the improvements and evolution of the security model in the ATLAS PanDA (Production and Distributed Analysis) system. We describe the security in the PanDA server which is in place to ensure that only authorized members of the VO are allowed to submit work into the system and that jobs are properly audited and monitored. We discuss the security in place between the pilot code itself and the PanDA server, ensuring that only properly authenticated workload is delivered to the pilot for execution. When the code to be executed is from a 'normal' ATLAS user, as opposed to the production system or other privileged actor, then the pilot may use an EGEE developed identity switching tool called gLExec. This changes the grid proxy available to the job and also switches the UNIX user identity to protect the privileges of the pilot code proxy. We describe the problems in using this system and how they are overcome. Finally, we discuss security drills which have been run using PanDA and show how these improved our operational security procedures.

  1. GEODSS Tracking Results on Asteroid 2012 DA14

    DTIC Science & Technology

    2013-06-12

    meteor until stony or metallic material remnants make it to the surface of the planet in which case they become meteorites . Theoretically, the...passing in close proximity to the planet are a daily manifestation. In fact within days of the 2012 DA14 transit of the planet a large meteorite entered...although meteorites approximately basketball size hit the Earth about once per day, and objects the size of an automobile reach us roughly on a weekly

  2. Da Vinci's codex and the anatomy of healthcare.

    PubMed

    Stephens-Borg, Keith

    2012-08-01

    We usually display a laid-back approach to medical jargon throughout our theatre work. The word 'perioperative' is built from the Greek word 'peri' (around) and the Latin 'operari' (to work). Latin and Greek became the prefixed language of choice for Leonardo da Vinci, and his research was pivotal in determining the way in which surgical procedures are documented. Ancient manuscripts aided the unfolding of the secrets of anatomy, and Leonardo revealed that art was the key in expressive detailed explanation.

  3. Is there any mantle plume beneath Tristan da Cunha?

    NASA Astrophysics Data System (ADS)

    Schloemer, A.; Geissler, W. H.; Jegen, M. D.; Jokat, W.

    2015-12-01

    Tristan da Cunha is a volcanic island in the South Atlantic located very close to the Mid-Atlantic Ridge. Generally, it is accepted to be the location of a mantle plume, which has been active at least since the breakup of Gondwana at 130 Ma, the time when the Paraná/Etendeka flood basalts were emplaced. Furthermore, it is associated with the formation of the Walvis Ridge and the Rio Grande Rise, and therefore it's one of the key examples of a hot spot track linking a flood basalt province to an active ocean island volcano. However, global tomography models are contradicting about the origin of Tristan da Cunha: Whether it is a deep mantle plume or caused by shallow plate tectonics. To gain a better understanding, we deployed 24 broadband ocean-bottom seismometers, 26 ocean-bottom electromagnetic stations and 2 seismological land stations in January 2012 with the German research vessel Maria S. Merian. We acquired continuous seismological data for one year and recovered the instruments in January 2013.We use cross-correlated travel time residuals of teleseismic earthquakes to perform a finite-frequency tomography to resolve the P wave velocity upper mantle structure beneath the island. Here we show our preliminary results of the 3-D velocity perturbations in the upper mantle: We do not image a plume-like structure directly beneath the island. Instead we observe a low velocity region in the southwest of our array that might be related to a local mantle upwelling (mantle plume). Additionally we show the local seismicity in the Tristan da Cunha region.Chen et al. and Baba et al. will present the first results on the magnetotelluric experiment and Ryberg et al. will present the crustal structure around the Tristan da Cunha hotspot.

  4. DA-9801 promotes neurite outgrowth via ERK1/2-CREB pathway in PC12 cells.

    PubMed

    Won, Jong Hoon; Ahn, Kyong Hoon; Back, Moon Jung; Ha, Hae Chan; Jang, Ji Min; Kim, Ha Hyung; Choi, Sang-Zin; Son, Miwon; Kim, Dae Kyong

    2015-01-01

    In the present study, we examined the mechanisms underlying the effect of DA-9801 on neurite outgrowth. We found that DA-9801 elicits its effects via the mitogen-activated protein kinase (MEK) extracellular signal-regulated kinase (ERK)1/2-cAMP response element-binding protein (CREB) pathway. DA-9801, an extract from a mixture of Dioscorea japonica and Dioscorea nipponica, was reported to promote neurite outgrowth in PC12 cells. The effects of DA-9801 on cell viability and expression of neuronal markers were evaluated in PC12 cells. To investigate DA-9801 action, specific inhibitors targeting the ERK signaling cascade were used. No cytotoxicity was observed in PC12 cells at DA-9801 concentrations of less than 30 µg/mL. In the presence of nerve growth factor (NGF, 2 ng/mL), DA-9801 promoted neurite outgrowth and increased the relative mRNA levels of neurofilament-L (NF-L), a marker of neuronal differentiation. The Raf-1 inhibitor GW5074 and MEK inhibitor PD98059 significantly attenuated DA-9801-induced neurite outgrowth. Additionally, the MEK1 and MEK2 inhibitor SL327 significantly attenuated the increase in the percentage of neurite-bearing PC12 cells induced by DA-9801 treatment. Conversely, the selective p38 mitogen-activated protein kinase inhibitor SB203580 did not attenuate the DA-9801 treatment-induced increase in the percentage of neurite-bearing PC12 cells. DA-9801 enhanced the phosphorylation of ERK1/2 and CREB in PC12 cells incubated with and without NGF. Pretreatment with PD98059 blocked the DA-9801-induced phosphorylation of ERK1/2 and CREB. In conclusion, DA-9801 induces neurite outgrowth by affecting the ERK1/2-CREB signaling pathway. Insights into the mechanism underlying this effect of DA-9801 may suggest novel potential strategies for the treatment of peripheral neuropathy.

  5. Domain annotation of trimeric autotransporter adhesins—daTAA

    PubMed Central

    Szczesny, Pawel; Lupas, Andrei

    2008-01-01

    Motivation: Trimeric autotransporter adhesins (TAAs), such as Yersinia YadA, Neisseria NadA, Moraxella UspAs, Haemophilus Hia and Bartonella BadA, are important pathogenicity factors of proteobacteria. Their high sequence diversity and distinct mosaic-like structure lead to difficulties in the annotation of their sequences. These stem from the large number of short repeats, the presence of compositionally unusual coiled-coils, fuzzy domain boundaries and regions of seemingly low sequence complexity. Results: We have developed a workflow, named daTAA, for the accurate domain annotation of TAAs. Its core consists of manually curated alignments and of knowledge-based rules that enhance assignments made by sequence similarity. Compared to general domain annotation servers such as PFAM, daTAA captures more domains and provides more sensitive domain detection, as well as integrated and detailed coiled-coil assignments. Availability: The daTAA server is freely accessible at http://toolkit.tuebingen.mpg.de/dataa Contact: andrei.lupas@tuebingen.mpg.de Supplementary information: Supplementary data are available at Bioinformatics online. PMID:18397894

  6. Common commercial cosmetic products induce arthritis in the DA rat.

    PubMed Central

    Sverdrup, B; Klareskog, L; Kleinau, S

    1998-01-01

    Many different agents, including mineral oil and silicone, have the capacity to act as immunological adjuvants, i.e., they can contribute to the activation of the immune system. Some adjuvants, including mineral oil, are known to induce arthritis in certain strains of rats after intradermal injection or percutaneous application. The aim of this study was to determine if common commercial cosmetic products containing mineral oil could induce arthritis in the highly susceptible DA (Dark Agouti) rat. Intradermal injection of five out of eight assayed cosmetic products without further additives resulted in arthritis with synovitis. One of the products induced a very aggressive arthritis, which had declined after 5-9 weeks. When this product was also assayed for arthritogenicity upon percutaneous administration, it induced a mild and transient arthritis in 5 out of 10 DA rats, whereas control animals showed no clinical signs of joint involvement. No arthritic reaction was seen in rats after peroral feeding with the most arthritogenic product or by intravaginal application of Freund's adjuvants. Silicone gel implants in DA rats did not cause arthritis. We conclude that mineral oils included in common commercially available products retain their adjuvant properties and are arthritogenic in the presently investigated arthritis-prone rat strain. There is yet no evidence that mineral oils present in cosmetics may contribute to arthritis in humans, but we suggest that this question should be subject to further investigation. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:9417771

  7. The DaVinci Group: a second modern Ophthalmotrope.

    PubMed

    Pruehsner, William R; Enderle, John D

    2006-01-01

    A group of undergraduate students at the University of Connecticut Biomedical Engineering Program has formed a "club" in order to more fully understand and educate themselves in modeling anatomical processes. This group is called the DaVinci Robot or DaVinci Group. Experiments to mechanically model the six extraocular muscles of the eye have been performed, each meeting little success. While researching methods that would lead to better success, the concept of the Ophthalmotrope was discovered. The Ophthalmotrope is a mechanical visual aide used in teaching the function of the extraocular muscles, prevalent in the mid 1800's. The Group decided to study this device and ultimately decided to build one. The paper presented here discusses our third experiment, currently under investigation, that is, to build an Opthalmotrope. Difficulties with this task are lack of any information with regard to how to construct this device. Presented are descriptions of the Group's initial experiments and research conducted into the construction of the Ophthalmotrpe. In the main body of the presented paper is a description of how the DaVinci Group Ophthalmotrope is constructed. Concluding is a discussion of the progress of the construction of the Ophthalmotrope along with a brief listing of research conducted in order to build the device.

  8. The ATLAS PanDA Pilot in Operation

    NASA Astrophysics Data System (ADS)

    Nilsson, P.; Caballero, J.; De, K.; Maeno, T.; Stradling, A.; Wenaus, T.; ATLAS Collaboration

    2011-12-01

    The Production and Distributed Analysis system (PanDA) [1-2] was designed to meet ATLAS [3] requirements for a data-driven workload management system capable of operating at LHC data processing scale. Submitted jobs are executed on worker nodes by pilot jobs sent to the grid sites by pilot factories. This paper provides an overview of the PanDA pilot [4] system and presents major features added in light of recent operational experience, including multi-job processing, advanced job recovery for jobs with output storage failures, gLExec [5-6] based identity switching from the generic pilot to the actual user, and other security measures. The PanDA system serves all ATLAS distributed processing and is the primary system for distributed analysis; it is currently used at over 100 sites worldwide. We analyze the performance of the pilot system in processing real LHC data on the OSG [7], EGI [8] and Nordugrid [9-10] infrastructures used by ATLAS, and describe plans for its evolution.

  9. Pictorial cues constrain depth in da Vinci stereopsis.

    PubMed

    Makino, Yoshinari; Yano, Masafumi

    2006-01-01

    "da Vinci stereopsis" is defined as depth seen in a monocular object occluded by a binocular one, and the visual system must solve its depth ambiguity [Nakayama, K., & Shimojo, S. (1990). da Vinci stereopsis: Depth and subjective occluding contours from unpaired image points. Vision Research, 30, 1811-1825]. Although fused images include various pictorial features, effects of pictorial depth cues have never been systematically investigated in da Vinci stereopsis. To examine this, we created stereograms consisting of a monocular bar flanked by binocular bars with a fixed large horizontal separation, in which the monocular bar induced a subjective occluding edge. Manipulating vertical size or contrast of the bars could affect the depth of the monocular bar. Conflicting these cues revealed that the effect of vertical size was stronger than that of contrast in all our subjects. Measurements of the depth indicated that the relative vertical size of the bars quantitatively determined the perceived depth, of which levels had large inter-subject differences. All these experiments indicate that the visual system can use the pictorial depth cues as a constraint to determine the depth of monocular elements.

  10. OpenDA-NEMO framework for ocean data assimilation

    NASA Astrophysics Data System (ADS)

    van Velzen, Nils; Altaf, Muhammad Umer; Verlaan, Martin

    2016-05-01

    Data assimilation methods provide a means to handle the modeling errors and uncertainties in sophisticated ocean models. In this study, we have created an OpenDA-NEMO framework unlocking the data assimilation tools available in OpenDA for use with NEMO models. This includes data assimilation methods, automatic parallelization, and a recently implemented automatic localization algorithm that removes spurious correlations in the model based on uncertainties in the computed Kalman gain matrix. We have set up a twin experiment where we assimilate sea surface height (SSH) satellite measurements. From the experiments, we can conclude that the OpenDA-NEMO framework performs as expected and that the automatic localization significantly improves the performance of the data assimilation algorithm by successfully removing spurious correlations. Based on these results, it looks promising to extend the framework with new kinds of observations and work on improving the computational speed of the automatic localization technique such that it becomes feasible to include large number of observations.

  11. Identification of an abundant 56 kDa protein implicated in food allergy as granule-bound starch synthase

    USDA-ARS?s Scientific Manuscript database

    Rice, the staple food of South and East Asian counties, is considered to be hypoallergenic. However, several clinical studies have documented rice-induced allergy in sensitive patients. Rice proteins with molecular weights of 14-16 kDa, 26 kDa, 33 kDa and 56 kDa have been identified as allergens. Re...

  12. [The Arabic influence in the "Colóquios dos simples e drogas da India" of Garcia da Orta].

    PubMed

    Ricordel, Joëlle

    2015-09-01

    The "Colóquios dos simples e drogas he cousas medicinais de Índia" (Conversations on the simples, drugs and medicinal substances of India) (1563) of Garcia da Orta is a botanical and pharmacognosy book. The author is a Portuguese physician who studied in the Spanish universities and practiced medicine mainly in India. He studies in short chapters presented in the form of dialogues about sixty simples. Sources to which he refers are indicative of a "classical" training, but also the mark of a curious and open mind to different cultures. The Arabic sources are numerous and mainly concern the identification of substances by abundant synonyms of their names in foreign languages and different medicinal uses that may have been done by the ancient physicians. However, Da Orta is critical with respect to these sources, seeking contradictions and differences of opinion among authors. He confronts them with the oral information collected thanks to a wide network of contacts.

  13. Seeking 'energy' vs. pain relief in spas in Brazil (Caldas da Imperatriz) and Portugal (Termas da Sulfurea).

    PubMed

    Quintela, Maria Manuel

    2011-04-01

    This paper is a comparative ethnography of the therapeutic practices at two different spa locations: Caldas da Imperatriz, SC, Brazil, and Termas da Sulfurea in Cabeco de Vide, Portugal. The comparison reveals the existence of contrasting 'explanatory models' held by the spa-goers as well as by the official medical systems. In the Portuguese context this model is highly medicalized; in the Brazilian case, spa treatments are viewed as 'alternative' or 'complementary' therapy and are also related to religious philosophies. Each model corresponds to a different idiom expressing certain experiences and world views, one focusing on 'pains' (dores) and the other on 'energy' (energia), the former leading to the rationale of 'curing', the latter to the notion of 'energizing'. In this paper the author intends to analyze and contrast the categories found in these models, which originate from different conceptions of health, illness and healing for Brazilian and Portuguese spa-goers.

  14. Characterization of a dopamine receptor (DA sub 2K ) in the kidney inner medulla

    SciTech Connect

    Huo, T.; Ye, M.Q.; Healy, D.P. )

    1991-04-15

    Dopamine (DA) produces a natriuretic/diuretic response in the kidney by mechanisms that are still not well understood. There is some indication that DA{sub 2} receptors may be involved in mediating the effects of DA, but little is known regarding the nature of this receptor in the kidney. Autoradiographic localization of ({sup 3}H)spiperone, a DA{sub 2} antagonist, indicated that high-density binding was restricted to inner medullary collecting ducts (IMCDs). ({sup 3}H)Spiperone binding was saturable, high affinity and high density. Functionally, DA stimulated prostaglandin E{sub 2} production by IMCD cells, an effect that could be blocked by the DA{sub 2} antagonist domperidone. These results indicate that the kidney inner medulla expresses a functional DA receptor that may represent a newly identified DA receptor subtype (here designated DA{sub 2K}). Moreover, these results suggest that the kidney inner medulla may be a significant site at which DA, either directly or indirectly, influences water and electrolyte excretion.

  15. High School Student's Alternative Conceptions About the Phenomenon of the Formation of the Moon Phases. (Spanish Title: Concepciones Alternativas de Alumnos de Educación Media Sobre el Fenómeno de Formación de las Fases de La Luna.) Concepções Alternativas de Alunos do Ensino Médio Sobre o Fenômeno de Formação das Fases da Lua

    NASA Astrophysics Data System (ADS)

    Iachel, Gustavo; Langhi, Rodolfo; Fernandes Scalvi, Rosa Maria

    2008-07-01

    Forty students, at ages between 14 and 18 years old, from three schools in Bauru city, were questioned about their alternative conceptions concerning the phenomenon of formation of the Moon Phases. It was observed that some of the pupils confound the phenomenon of the formation of the Moon Phases with the phenomenon of the formation of the lunar eclipses, others are unaware of the reason of the phenomenon, they present incoherent alternative conceptions of the reality or incomplete conceptions. The results found here are aimed at the teachers of Elementary Education and can be used as a subsidy for future development of new pedagogical methods. Cuarenta estudiantes, con edad entre 14 y 18 años, pertenecientes a tres escuelas de la ciudad de Bauru, fueron cuestionados sobre sus concepciones alternativas acerca del fenómeno de la formación de las fases de la Luna. Fue observado que algunos alumnos confunden el fenómeno de formación de las fases de la Luna con el fenómeno de formación de los eclipses lunares, otros desconocen el motivo delfenómeno, presentan concepciones alternativas incoherentes con la realidad o bien presentan concepciones incompletas. Los resultados aquí encontrados son destinados a los profesores de la Enseñanza Básica y podrán ser usados como ayuda para el futuro desarrollo de nuevos métodos pedagógicos. Quarenta estudantes, com idades entre 14 e 18 anos, pertencentes a três escolas da cidade de Bauru, foram questionados sobre suas concepções alternativas acerca do fenômeno de formação das fases da Lua. Foi observado que alguns dos alunos confundem o fenômeno da formação das fases da Lua com o fenômeno da formação dos eclipses lunares, outros desconhecem o motivo do fenômeno, apresentam concepções alternativas incoerentes com a realidade ou então concepções incompletas. Os resultados aqui encontrados são destinados aos professores do Ensino Básico e poderão ser usados como subsídio parafuturo desenvolvimento de

  16. The primary structure of skeletal muscle myosin heavy chain: III. Sequence of the 22 kDa fragment and the alignment of the 23 kDa, 50 kDa, and 22 kDa fragments.

    PubMed

    Maita, T; Miyanishi, T; Matsuzono, K; Tanioka, Y; Matsuda, G

    1991-07-01

    The amino acid sequence of the 197-residue 22 kDa fragment from chicken pectoralis muscle was determined to be as follows: K-K-G-S-S-F-Q-T-V-S-A-L-F-R-E-N-L-N-K-L- M-A-N-L-R-S-T-H-P-H-F-V-R-C-I-I-P-N-E-T-K-T-P-G-A-M-E-H-E-L-V-L-H-Q-L-R- C-N-G-V- L-E-G-I-R-I-C-R-K-G-F-P-S-R-V-L-Y-A-D-F-K-Q-R-Y-R-V-L-N-A-S-A-I-P-E-G-Q- F-M-D-S- K-K-A-S-E-K-L-L-G-S-I-D-V-D-h-T-Q-Y-R-F-G-H-T-K-V-F-F-K-A-G-L-L-G-L-L-E- E-M-R-D- D-K-L-A-E-I-I-T-R-T-Q-A-R-C-R-G-F-L-M-R-V-E-Y-R-R-M-V-E-R-R-E-S-I-F-C-I- Q-Y-N-V-R-S-F-M-N-V-K-H-W-P-W-M-K-L-F-F-K, where h stands for 3-N-methylhistidine. The amino acid sequences of the 22 kDa fragment and its equivalent fragment from chicken ventricle and gizzard muscle myosins were also determined by our group. Predicted secondary structures of these 22 kDa fragment regions and of the reported chicken embryo myosin revealed some possible structural differences.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Leonardo da Vinci and Kethem-Kiveris vena.

    PubMed

    Dolezal, Antonín; Skorepova-Honzlova, Zita; Jelen, Karel

    2012-01-01

    In the drawing of coitus by Leonardo da Vinci are pictured the contemporary hypotheses regarding this act. The authors analyze the mamillaruteral connection depicted by the artist and grow up to believe that this is a hypothetical kiveris vena, female vein described by Anatomist Master Nicolai Physicus from the Salerno School. The Hebrew roots were found in the name. The connection is described also by Mondino in The Anathomia. The same connection can be found in the picture of the pregnant woman in Fasciculus Medicinæ by Johannes De Ketham.

  18. LEONARDO DA VINCI AND THE ORIGIN OF SEMEN.

    PubMed

    Noble, Denis; DiFrancesco, Dario; Zancani, Diego

    2014-12-20

    It is well known that Leonardo da Vinci made several drawings of the human male anatomy. The early drawings (before 1500) were incorrect in identifying the origin of semen, where he followed accepted teaching of his time. It is widely thought that he did not correct this mistake, a view that is reflected in several biographies. In fact, he made a later drawing (after 1500) in which the description of the anatomy is remarkably accurate and must have been based on careful dissection. In addition to highlighting this fact, acknowledged previously in only one other source, this article reviews the background to Leonardo's knowledge of the relevant anatomy.

  19. A quasi-passive CMOS pipeline D/A converter

    NASA Technical Reports Server (NTRS)

    Wang, Fong-Jim; Temes, Gabor C.; Law, Simon

    1989-01-01

    A novel pipeline digital-to-analog converter configuration, based on switched-capacitor techniques, is described. An n-bit D/A conversion can be implemented by cascading n + 1 unit cells. The device count of the circuit increases linearly, not exponentially, with the conversion accuracy. The new configuration can be pipelined. Hence, the conversion rate can be increased without requiring a higher clock rate. An experimental 10-bit DAC prototype has been fabricated using a 3-micron CMOS process. The results show that high-speed, high-accuracy, and low-power operation can be achieved without special process or postprocess trimming.

  20. Ultratight crystal packing of a 10 kDa protein

    SciTech Connect

    Trillo-Muyo, Sergio; Chruszcz, Maksymilian; Minor, Wladek; Kuisiene, Nomeda

    2013-03-01

    The crystal structure of the C-terminal domain of a putative U32 peptidase from G. thermoleovorans is reported; it is one of the most tightly packed protein structures reported to date. While small organic molecules generally crystallize forming tightly packed lattices with little solvent content, proteins form air-sensitive high-solvent-content crystals. Here, the crystallization and full structure analysis of a novel recombinant 10 kDa protein corresponding to the C-terminal domain of a putative U32 peptidase are reported. The orthorhombic crystal contained only 24.5% solvent and is therefore among the most tightly packed protein lattices ever reported.

  1. Leonardo da Vinci and the origin of semen

    PubMed Central

    Noble, Denis; DiFrancesco, Dario; Zancani, Diego

    2014-01-01

    It is well known that Leonardo da Vinci made several drawings of the human male anatomy. The early drawings (before 1500) were incorrect in identifying the origin of semen, where he followed accepted teaching of his time. It is widely thought that he did not correct this mistake, a view that is reflected in several biographies. In fact, he made a later drawing (after 1500) in which the description of the anatomy is remarkably accurate and must have been based on careful dissection. In addition to highlighting this fact, acknowledged previously in only one other source, this article reviews the background to Leonardo's knowledge of the relevant anatomy. PMID:27494016

  2. High Resolution EUV & FUV Spectroscopy of DA White Dwarfs

    NASA Astrophysics Data System (ADS)

    Barstow, M. A.; Good, S. A.; Bannister, N. P.; Burleigh, M. R.; Holberg, J. B.; Bruhweiler, F. C.; Napiwotzki, R.; Cruddace, R. G.; Kowalski, M. P.

    We report on recent results from a high-resolution spectroscopic survey of hot DA white dwarfs, based on IUE, FUSE and HST observations. For the first time, we address the measurement of element abundances in a completely objective manner with a spectroscopic model fitting technique, which allows us to consider formally the limits that can be placed on abundances in stars where no heavy elements are detected. We also include our latest analysis of the high resolution EUV spectrum of G191-B2B recorded by J-PEX.

  3. Mammary artery harvesting using the Da Vinci Si robotic system

    PubMed Central

    Canale, Leonardo Secchin; Bonatti, Johannes

    2014-01-01

    Internal mammary artery harvesting is an essential part of any coronary artery bypass operation. Totally endoscopic coronary artery bypass graft surgery has become reality in many centers as a safe and effective alternative to conventional surgery in selected patients. Internal mammary artery harvesting is the initial part of the procedure and should be performed equally safely if one wants to achieve excellence in patency rates for the bypass. We here describe the technique for mammary harvesting with the Da Vinci Si robotic system. PMID:24896171

  4. Visual tracking of da Vinci instruments for laparoscopic surgery

    NASA Astrophysics Data System (ADS)

    Speidel, S.; Kuhn, E.; Bodenstedt, S.; Röhl, S.; Kenngott, H.; Müller-Stich, B.; Dillmann, R.

    2014-03-01

    Intraoperative tracking of laparoscopic instruments is a prerequisite to realize further assistance functions. Since endoscopic images are always available, this sensor input can be used to localize the instruments without special devices or robot kinematics. In this paper, we present an image-based markerless 3D tracking of different da Vinci instruments in near real-time without an explicit model. The method is based on different visual cues to segment the instrument tip, calculates a tip point and uses a multiple object particle filter for tracking. The accuracy and robustness is evaluated with in vivo data.

  5. An improvement of LLNA:DA to assess the skin sensitization potential of chemicals.

    PubMed

    Zhang, Hongwei; Shi, Ying; Wang, Chao; Zhao, Kangfeng; Zhang, Shaoping; Wei, Lan; Dong, Li; Gu, Wen; Xu, Yongjun; Ruan, Hongjie; Zhi, Hong; Yang, Xiaoyan

    2017-01-01

    We developed a modified local lymph node assay based on ATP (LLNA:DA), termed the Two-Stage LLNA:DA, to further reduce the animal numbers in the identification of sensitizers. In the Two-Stage LLNA:DA procedure, 13 chemicals ranging from non-sensitizers to extreme sensitizers were selected. The first stage used reduced LLNA:DA (rLLNA:DA) to screen out sensitive chemicals. The second stage used LLNA:DA based on OECD 442 (A) to classify those potential sensitizers screened out in the first stage. In the first stage, the SIs of the methyl methacrylate, salicylic acid, methyl salicylate, ethyl salicylate, isopropanol and propanediol were below 1.8 and need not to be tested in the second step. Others continued to be tested by LLNA:DA. In the second stage, sodium lauryl sulphate and xylene were classified as weak sensitizers. a-hexyl cinnamic aldehyde and eugenol were moderate sensitizers. Benzalkonium chloride and glyoxal were strong sensitizers, and phthalic anhydride was an extreme sensitizer. The 9/9, 11/12, 10/11, and 8/13 (positive or negative only) categories of the Two-Stage LLNA:DA were consistent with those from the other methods (LLNA, LLNA:DA, GPMT/BT and HMT/HPTA), suggesting that Two-Stage LLNA:DA have a high coincidence rate with reported data. In conclusion, The Two-Stage LLNA:DA is in line with the "3R" rules, and can be a modification of LLNA:DA but needs more study.

  6. Purification, characterization and gene cloning of Da-36, a novel serine protease from Deinagkistrodon acutus venom.

    PubMed

    Zheng, Ying; Ye, Feng-Ping; Wang, Jie; Liao, Guo-Yang; Zhang, Yun; Fan, Quan-Shui; Lee, Wen-Hui

    2013-06-01

    A serine protease termed Da-36 was isolated from crude venom of Deinagkistrodon acutus. The enzyme was a single chain protein with an apparent molecular weight of 36,000 on SDS-PAGE with an isoelectric point of 6.59. Da-36 could clot human plasma by cleaving the Aα, Bβ and γ chains of fibrinogen and also exhibited arginine esterase activity. The proteolytic activity of Da-36 toward TAME was strongly inhibited by PMSF and moderately affected by benzamidine and aprotinin, indicating that it was a serine protease. Meanwhile, Da-36 showed stability with wide temperature (20-50 °C) and pH value ranges (pH 6-10). Divalent metal ions of Ca(2+), Mg(2+), and Mn(2+) had no effects but Zn(2+) and Cu(2+) inhibited the arginine esterase activity of Da-36. Total DNA was extracted directly from the lyophilized crude venom and the gene (5.5 kbp) coding for Da-36 had been successfully cloned. Sequence analysis revealed that the Da-36 gene contained five exons and four introns. The mature Da-36 was encoded by four separate exons. The deduced mature amino acid sequence of Da-36 was in good agreement with the determined N-terminal sequence of the purified protein and shared high homology with other serine proteases isolated from different snake venoms. Blast search using amino acid sequence of Da-36 against public database revealed that Da-36 showed a maximal identity of 90% with both Dav-X (Swiss-Prot: Q9I8W9.1) and thrombin-like protein 1 (GenBank: AAW56608.1) from the same snake species, indicating that Da-36 is a novel serine protease.

  7. Identification of Methylated Deoxyadenosines in Genomic DNA by dA6m DNA Immunoprecipitation

    PubMed Central

    Koziol, Magdalena J.; Bradshaw, Charles R.; Allen, George E.; Costa, Ana S.H.; Frezza, Christian

    2017-01-01

    dA6m DNA immunoprecipitation followed by deep sequencing (DIP-Seq) is a key tool in identifying and studying the genome-wide distribution of N6-methyldeoxyadenosine (dA6m). The precise function of this novel DNA modification remains to be fully elucidated, but it is known to be absent from transcriptional start sites and excluded from exons, suggesting a role in transcriptional regulation (Koziol et al., 2015). Importantly, its existence suggests that DNA might be more diverse than previously believed, as further DNA modifications might exist in eukaryotic DNA (Koziol et al., 2015). This protocol describes the method to perform dA6m DNA immunoprecipitation (DIP), as was applied to characterize the first dA6m methylome analysis in higher eukaryotes (Koziol et al., 2015). In this protocol, we describe how genomic DNA is isolated, fragmented and then DNA containing dA6m is pulled down with an antibody that recognizes dA6m in genomic DNA. After subsequent washes, DNA fragments that do not contain dA6m are eliminated, and the dA6m containing fragments are eluted from the antibody in order to be processed further for subsequent analyses. Background This protocol was developed in order to identify regions in the genome that contain dA6m. It can be used to detect dA6m in different genomes. As a guideline, this protocol was established from existing approaches used to detect adenosine methylation in RNA (Dominissini et al., 2013). We developed this protocol and adapted it for the detection of dA6m in DNA, rather than detecting adenosine methylation RNA. This was required, as no protocol was available at that time to allow the genome-wide identification of dA6m in eukaryotic DNA. PMID:28180135

  8. A 45-kDa acetylcholinesterase protoxin of Aeromonas hydrophila: purification and immunogenicity in fish.

    PubMed

    Pérez, M J; Rodríguez, L A; Fernández-Briera, A; Nieto, T P

    2002-05-21

    A rabbit antiserum to the 15-kDa acetylcholinesterase toxin neutralised the lethal effect of the 15-kDa toxin of Aeromonas hydrophila when injected into trout. However, immunisation of fish with the 15-kDa toxoid failed to induce an antibody response, and a higher molecular mass form of this toxin was purified from the extracellular products with the aim of inducing an immune response in fish. The optimal conditions for production of extracellular products by A. hydrophila strain B32 were studied to increase the concentration of this protoxin. The extracellular products were fractionated by molecular exclusion chromatography to yield a purified protoxin with an estimated molecular mass of 45 kDa by SDS-PAGE and which gave a positive reaction in Western blotting with the rabbit anti-15-kDa toxin serum. Since the 45-kDa protoxin showed lower specific acetylcholinesterase activity than the active 15-kDa toxin, the behaviour of the active site was studied using specific inhibitors. This 45-kDa protoxin was 13.3-fold less toxic than the 15-kDa toxin and induced antibody production in fish.

  9. Evolution of the ATLAS PanDA workload management system for exascale computational science

    NASA Astrophysics Data System (ADS)

    Maeno, T.; De, K.; Klimentov, A.; Nilsson, P.; Oleynik, D.; Panitkin, S.; Petrosyan, A.; Schovancova, J.; Vaniachine, A.; Wenaus, T.; Yu, D.; Atlas Collaboration

    2014-06-01

    An important foundation underlying the impressive success of data processing and analysis in the ATLAS experiment [1] at the LHC [2] is the Production and Distributed Analysis (PanDA) workload management system [3]. PanDA was designed specifically for ATLAS and proved to be highly successful in meeting all the distributed computing needs of the experiment. However, the core design of PanDA is not experiment specific. The PanDA workload management system is capable of meeting the needs of other data intensive scientific applications. Alpha-Magnetic Spectrometer [4], an astro-particle experiment on the International Space Station, and the Compact Muon Solenoid [5], an LHC experiment, have successfully evaluated PanDA and are pursuing its adoption. In this paper, a description of the new program of work to develop a generic version of PanDA will be given, as well as the progress in extending PanDA's capabilities to support supercomputers and clouds and to leverage intelligent networking. PanDA has demonstrated at a very large scale the value of automated dynamic brokering of diverse workloads across distributed computing resources. The next generation of PanDA will allow other data-intensive sciences and a wider exascale community employing a variety of computing platforms to benefit from ATLAS' experience and proven tools.

  10. Effects of DA-Phen, a dopamine-aminoacidic conjugate, on alcohol intake and forced abstinence.

    PubMed

    Sutera, Flavia Maria; De Caro, Viviana; Cannizzaro, Carla; Giannola, Libero Italo; Lavanco, Gianluca; Plescia, Fulvio

    2016-09-01

    The mesolimbic dopamine (DA) system plays a key role in drug reinforcement and is involved in the development of alcohol addiction. Manipulation of the DAergic system represents a promising strategy to control drug-seeking behavior. Previous studies on 2-amino-N-[2-(3,4-dihydroxy-phenyl)-ethyl]-3-phenyl-propionamide (DA-Phen) showed in vivo effects as a DA-ergic modulator. This study was aimed at investigate DA-Phen effects on operant behavior for alcohol seeking behavior, during reinstatement following subsequent periods of alcohol deprivation. For this purpose, male Wistar rats were tested in an operant paradigm of self-administration; behavioral reactivity and anxiety like-behavior during acute abstinence were evaluated. A characterization of DA-Phen CNS targeting by its quantification in the brain was also carried out. Our findings showed that DA-Phen administration was able to reduce relapse in alcohol drinking by 50% and reversed the alterations in behavioral reactivity and emotionality observed during acute abstinence. In conclusion, DA-Phen can reduce reinstatement of alcohol drinking in an operant-drinking paradigm following deprivation periods and reverse abstinence-induced behavioral phenotype. DA-Phen activity seems to be mediated by the modulation of the DAergic transmission. However further studies are needed to characterize DA-Phen pharmacodynamic and pharmacokinetic properties, and its potential therapeutic profile in alcohol addiction.

  11. PX-52, A novel inhibitor of 14 kDa secretory and 85 kDa cytosolic phospholipases A2.

    PubMed

    Franson, R C; Rosenthal, M D

    1997-01-01

    Previously we reported that PGBx, a prostaglandin oligomer with anti-inflammatory activity, inhibited 14 kDa phospholipase A2 (PLA2) activity and blocked arachidonic acid mobilization in prelabeled human neutrophils (Biochim. Biophys. Acta 1006:272-277, 278-286, 1989) This study describes a new inhibitor of phospholipase A2, PX-52, that also blocks agonist induced arachidonic acid mobilization in prelabeled cells. PX-52, a fatty acid polymer, inhibited hydrolysis of 14C-oleate labeled E.coli by a variety of 14 kDa PLA2s including human PMN, sperm, synovial fluid and disc, as well as porcine pancreas, N. naja, and bee venom in a dose-dependent manner with IC50s ranging from 1.0-3.7 uM. Inhibition of activity was comparable at different Ca2+ concentrations, but was relieved by increasing substrate concentration or by methylation of PX-52. Hydrolysis of [14C]-arachidonyl phosphatidylcholine by 85 kDa, cytosolic PLA2 from U937 cells was similarly inhibited by PX-52, the IC50 = 5 uM. Arachidonic acid mobilization induced by A23187 in prelabeled human PMNs was blocked by PX-52; IC50 = 10-15 uM while concentrations of up to 80 uM oleate had no effect. These results demonstrate that PX-52 inhibits the in vitro activity of secretory and cytosolic PLA2s and agonist-induced arachidonic acid release from human cells. Given its ability to block the arachidonic acid cascade, PX-52 may be useful in the control of inflammation.

  12. Purification and characterization of human 72-kDa gelatinase (type IV collagenase). Use of immunolocalisation to demonstrate the non-coordinate regulation of the 72-kDa and 95-kDa gelatinases by human fibroblasts.

    PubMed

    Hipps, D S; Hembry, R M; Docherty, A J; Reynolds, J J; Murphy, G

    1991-04-01

    Human gingival fibroblast gelatinase (type IV collagenase) has been purified to homogeneity using a combination of ion exchange chromatography, gel filtration and affinity chromatography. The purified proenzyme electrophoresed under reducing conditions as a single band of 72 kDa which could be activated to a species of 65 kDa. Gelatinase was activated by organomercurials by a process apparently initiated by a conformational change and involving self-cleavage. It was not activated by trypsin or plasmin unlike the other family members, collagenase and stromelysin. Gelatinase otherwise exhibited properties typical of the metalloproteinases: it was inhibited by metal chelating agents and by the specific inhibitor TIMP (tissue inhibitor of metalloproteinases). Its major substrate was shown to be denatured collagen although it was also able to degrade native type IV and V collagens. A polyclonal antibody was raised in a sheep using the purified enzyme as antigen. The antiserum recognised and specifically inhibited the 72-kDa gelatinase but not a 95-kDa gelatinase from pig leukocytes. It was used in immunolocalisation studies on human fibroblasts to investigate the regulation of the production of the two Mr forms of gelatinase. These studies clearly demonstrate that human fibroblasts constitutively synthesize and secrete 72-kDa gelatinase but that 95-kDa gelatinase was inducible by agents such as cytokines. The significance of these results in relation to the likely in vivo rôle of gelatinases is discussed.

  13. The complex impact structure Serra da Cangalha, Tocantins State, Brazil

    NASA Astrophysics Data System (ADS)

    Kenkmann, Thomas; Vasconcelos, Marcos A. R.; Crósta, Alvaro P.; Reimold, Wolf U.

    2011-06-01

    Serra da Cangalha is a complex impact structure with a crater diameter of 13,700 m and a central uplift diameter of 5800 m. New findings of shatter cones, planar fractures, feather features, and possible planar deformation features are presented. Several ring-like features that are visible on remote sensing imagery are caused by selective erosion of tilted strata. The target at Serra da Cangalha is composed of Devonian to Permian sedimentary rocks, mainly sandstones that are interlayered with siltstone and claystones. NNE-SSW and WNW-ESE-striking joint sets were present prior to the impact and also overprinted the structure after its formation. As preferred zones of weakness, these joint sets partly controlled the shape of the outer perimeter of the structure and, in particular, affected the deformation within the central uplift. Joints in radial orientation to the impact center did not undergo a change in orientation during tilting of strata when the central uplift was formed. These planes were used as major displacement zones. The asymmetry of the central uplift, with preferred overturning of strata in the northern to western sector, may suggest a moderately oblique impact from a southerly direction. Buckle folding of tilted strata, as well as strata overturning, indicates that the central uplift became gravitationally unstable at the end of crater formation.

  14. Chandra Confirmation of a Pulsar Wind Nebula in DA 495

    NASA Technical Reports Server (NTRS)

    Arzoumanian, Z.; Safi-Harb, S.; Landecker, T.L.; Kothes, R.; Camilo, F.

    2008-01-01

    As part of a multiwavelength study of the unusual radio supernova remnant DA 495, we present observations made with the Chandra X-ray Observatory. Imaging and spectroscopic analysis confirms the previously detected X-ray source at the heart of the annular radio nebula, establishing the radiative properties of two key emission components: a soft unresolved source with a blackbody temperature of 1 MK consistent with a neutron star, surrounded by a nontherma1 nebula 40" in diameter exhibiting a power-law spectrum with photon index Gamma = 1.63, typical of a pulsar wind nebula. Morphologically, the nebula appears to be slightly extended along a direction, in projection on the sky, previously demonstrated to be of significance in radio and ASCA observations; we argue that this represents the orientation of the pulsar spin axis. At smaller scales, a narrow X-ray feature is seen extending out 5" from the point source, but energetic arguments suggest that it is not the resolved termination shock of the pulsar wind against the ambient medium. Finally, we argue based on synchrotron lifetimes in the nebular magnetic field that DA 495 represents the first example of a pulsar wind nebula in which electromagnetic flux makes up a significant part, together with particle flux, of the neutron star's wind.

  15. Calibration of Synthetic Photometry Using DA White Dwarfs

    NASA Astrophysics Data System (ADS)

    Holberg, J. B.; Bergeron, Pierre

    2006-09-01

    We have calibrated four major ground-based photometric systems with respect to the Hubble Space Telescope (HST) absolute flux scale, which is defined by Vega and four fundamental DA white dwarfs. These photometric systems include the Johnson-Kron-Cousins UBVRI, the Strömgren uvby filters, the Two Micron All Sky Survey JHKs, and the Sloan Digital Sky Survey ugriz filters. Synthetic magnitudes are calculated from model white dwarf spectra folded through the published filter response functions; these magnitudes in turn are absolutely calibrated with respect to the HST flux scale. Effective zero-magnitude fluxes and zero-point offsets of each system are determined. In order to verify the external observational consistency, as well as to demonstrate the applicability of these definitions, the synthetic magnitudes are compared with the respective observed magnitudes of larger sets of DA white dwarfs that have well-determined effective temperatures and surface gravities and span a wide range in both of these parameters.

  16. D-A and D-2 dopamine receptor function in the rabbit retina: a model for the central nervous system

    SciTech Connect

    Hensler, J.G.

    1987-01-01

    Studies were done investigating the effect of the synaptic concentration of the transmitter DA, modified by changes in the frequency of electrical field stimulation and by the DA uptake inhibitor nomifensine, on the modulation of /sup 3/H-DA release by D-2 DA autoreceptors and by melatonin receptor sites. At lower synaptic concentrations of the transmitter dopamine, D-2 DA receptor agonists were more potent, while antagonists were more potent when the synaptic concentration of transmitter was higher. The potency of melatonin to inhibit DA release was not altered by the frequency of field stimulation of by frequency-dependent changes in the synaptic concentration of the transmitter.

  17. Axonal patterns and targets of dA1 interneurons in the chick hindbrain.

    PubMed

    Kohl, Ayelet; Hadas, Yoav; Klar, Avihu; Sela-Donenfeld, Dalit

    2012-04-25

    Hindbrain dorsal interneurons that comprise the rhombic lip relay sensory information and coordinate motor outputs. The progenitor dA1 subgroup of interneurons, which is formed along the dorsal-most region of the caudal rhombic lip, gives rise to the cochlear and precerebellar nuclei. These centers project sensory inputs toward upper-brain regions. The fundamental role of dA1 interneurons in the assembly and function of these brainstem nuclei is well characterized. However, the precise en route axonal patterns and synaptic targets of dA1 interneurons are not clear as of yet. Novel genetic tools were used to label dA1 neurons and trace their axonal trajectories and synaptic connections at various stages of chick embryos. Using dA1-specific enhancers, two contralateral ascending axonal projection patterns were identified; one derived from rhombomeres 6-7 that elongated in the dorsal funiculus, while the other originated from rhombomeres 2-5 and extended in the lateral funiculus. Targets of dA1 axons were followed at later stages using PiggyBac-mediated DNA transposition. dA1 axons were found to project and form synapses in the auditory nuclei and cerebellum. Investigation of mechanisms that regulate the patterns of dA1 axons revealed a fundamental role of Lim-homeodomain (HD) proteins. Switch in the expression of the specific dA1 Lim-HD proteins Lhx2/9 into Lhx1, which is typically expressed in dB1 interneurons, modified dA1 axonal patterns to project along the routes of dB1 subgroup. Together, the results of this research provided new tools and knowledge to the assembly of trajectories and connectivity of hindbrain dA1 interneurons and of molecular mechanisms that control these patterns.

  18. Immunoreactivity of the Mycobacterium avium subsp. paratuberculosis 19-kDa lipoprotein

    PubMed Central

    Huntley, Jason FJ; Stabel, Judith R; Bannantine, John P

    2005-01-01

    Background The Mycobacterium tuberculosis 19-kDa lipoprotein has been reported to stimulate both T and B cell responses as well as induce a number of Th1 cytokines. In order to evaluate the Mycobacterium avium subsp. paratuberculosis (M. avium subsp. paratuberculosis) 19-kDa lipoprotein as an immunomodulator in cattle with Johne's disease, the gene encoding the 19-kDa protein (MAP0261c) was analyzed. Results MAP0261c is conserved in mycobacteria, showing a 95% amino acid identity in M. avium subspecies avium, 84% in M. intracellulare and 76% in M. bovis and M. tuberculosis. MAP0261c was cloned, expressed, and purified as a fusion protein with the maltose-binding protein (MBP-19 kDa) in Escherichia coli. IFN-γ production was measured from 21 naturally infected and 9 control cattle after peripheral blood mononuclear cells (PBMCs) were stimulated with a whole cell lysate (WCL) of M. avium subsp. paratuberculosis or the recombinant MBP-19 kDa. Overall, the mean response to MBP-19 kDa was not as strong as the mean response to the WCL. By comparison, cells from control, non-infected cattle did not produce IFN-γ after stimulation with either WCL or MBP-19 kDa. To assess the humoral immune response to the 19-kDa protein, sera from cattle with clinical Johne's disease were used in immunoblot analysis. Reactivity to MBP-19 kDa protein, but not MBP alone, was observed in 9 of 14 infected cattle. Antibodies to the 19-kDa protein were not observed in 8 of 9 control cows. Conclusions Collectively, these results demonstrate that while the 19-kDa protein from M. avium subsp. paratuberculosis stimulates a humoral immune response and weak IFN-γ production in infected cattle, the elicited responses are not strong enough to be used in a sensitive diagnostic assay. PMID:15663791

  19. The 37kDa/67kDa Laminin Receptor acts as a receptor for Aβ42 internalization

    PubMed Central

    Da Costa Dias, Bianca; Jovanovic, Katarina; Gonsalves, Danielle; Moodley, Kiashanee; Reusch, Uwe; Knackmuss, Stefan; Weinberg, Marc S.; Little, Melvyn; Weiss, Stefan F. T.

    2014-01-01

    Neuronal loss is a major neuropathological hallmark of Alzheimer's disease (AD). The associations between soluble Aβ oligomers and cellular components cause this neurotoxicity. The 37 kDa/67 kDa laminin receptor (LRP/LR) has recently been implicated in Aβ pathogenesis. In this study the mechanism underlying the pathological role of LRP/LR was elucidated. Försters Resonance Energy Transfer (FRET) revealed that LRP/LR and Aβ form a biologically relevant interaction. The ability of LRP/LR to form stable associations with endogenously shed Aβ was confirmed by pull down assays and Aβ-ELISAs. Antibody blockade of this association significantly lowered Aβ42 induced apoptosis. Furthermore, antibody blockade and shRNA mediated downregulation of LRP/LR significantly hampered Aβ42 internalization. These results suggest that LRP/LR is a receptor for Aβ42 internalization, mediating its endocytosis and contributing to the cytotoxicity of the neuropeptide by facilitating intra-cellular Aβ42 accumulation. These findings recommend anti-LRP/LR specific antibodies and shRNAs as potential therapeutic tools for AD treatment. PMID:24990253

  20. Tdp studies and tests for C. A. Energia Electrica de Venezuela (enelven) at planta ramon laguna, units RL-17 and RL-10. Volume 3. Unit RL-10 turbine generator condition assessment report and units RL-10 and RL-11 boiler feed pump conditon assessment report. Export trade information

    SciTech Connect

    Not Available

    1991-03-28

    The study, conducted by Babcock and Wilcox, was funded by the U.S. Trade and Development agency on behalf of Enelven. In order to maximize generated power output and minimize operating costs at Planta Ramon Laguna, tests were done to evaluate the condition of equipment at the plant. In order to identify any damage and determine the operating output of each unit, assessments were done of the furnaces, boilers, generators and boiler feed pumps being used in the plant. The report presents the results of these tests. This is the last of three volumes and it is divided into the following sections: (1) Condition Assessment of Unit RL-10 Turbine-Generator; (2) Condition Assessment of Unit RL-10 and RL-11 Boiler Feed Pumps.

  1. Foxa2 acts as a co-activator potentiating expression of the Nurr1-induced DA phenotype via epigenetic regulation.

    PubMed

    Yi, Sang-Hoon; He, Xi-Biao; Rhee, Yong-Hee; Park, Chang-Hwan; Takizawa, Takumi; Nakashima, Kinichi; Lee, Sang-Hun

    2014-02-01

    Understanding how dopamine (DA) phenotypes are acquired in midbrain DA (mDA) neuron development is important for bioassays and cell replacement therapy for mDA neuron-associated disorders. Here, we demonstrate a feed-forward mechanism of mDA neuron development involving Nurr1 and Foxa2. Nurr1 acts as a transcription factor for DA phenotype gene expression. However, Nurr1-mediated DA gene expression was inactivated by forming a protein complex with CoREST, and then recruiting histone deacetylase 1 (Hdac1), an enzyme catalyzing histone deacetylation, to DA gene promoters. Co-expression of Nurr1 and Foxa2 was established in mDA neuron precursor cells by a positive cross-regulatory loop. In the presence of Foxa2, the Nurr1-CoREST interaction was diminished (by competitive formation of the Nurr1-Foxa2 activator complex), and CoREST-Hdac1 proteins were less enriched in DA gene promoters. Consequently, histone 3 acetylation (H3Ac), which is responsible for open chromatin structures, was strikingly increased at DA phenotype gene promoters. These data establish the interplay of Nurr1 and Foxa2 as the crucial determinant for DA phenotype acquisition during mDA neuron development.

  2. Tuning PEG-DA hydrogel properties via solvent-induced phase separation (SIPS)†

    PubMed Central

    Bailey, Brennan Margaret; Hui, Vivian; Fei, Ruochong

    2012-01-01

    Poly(ethylene glycol) diacrylate (PEG-DA) hydrogels are widely utilized to probe cell-material interactions and ultimately for a material-guided approach to tissue regeneration. In this study, PEG-DA hydrogels were fabricated via solvent-induced phase separation (SIPS) to obtain hydrogels with a broader range of tunable physical properties including morphology (e.g. porosity), swelling and modulus (G′). In contrast to conventional PEG-DA hydrogels prepared from an aqueous precursor solution, the reported SIPS protocol utilized a dichloromethane (DCM) precursor solution which was sequentially photopolymerized, dried and hydrated. Physical properties were further tailored by varying the PEG-DA wt% concentration (5 wt%–25 wt%) and Mn (3.4k and 6k g mol −1). SIPS produced PEG-DA hydrogels with a macroporous morphology as well as increased G′ values versus the corresponding conventional PEG-DA hydrogels. Notably, since the total swelling was not significantly changed versus the corresponding conventional PEG-DA hydrogels, pairs or series of hydrogels represent scaffolds in which morphology and hydration or G′ and hydration are uncoupled. In addition, PEG-DA hydrogels prepared via SIPS exhibited enhanced degradation rates. PMID:22956857

  3. 40 CFR 60.44Da - Standard for nitrogen oxides (NOX).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Standard for nitrogen oxides (NOX). 60.44Da Section 60.44Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS... for nitrogen oxides (NOX). (a) On and after the date on which the initial performance test is...

  4. Paśu Ayurvĕda (veterinary medicine) in Garudapurăņa.

    PubMed

    Varanasi, Subhose; Narayana, A

    2007-01-01

    The history of veterinary medicine is closely tied to the development of human medicine. Evidence of animal medicine has been found in ancient civilizations, such as those of the Hindu, Babylonians, Hebrews, Arabs, Greeks, and Romans. Ancient Indian literature in the form of the holy Vĕda, Purăna, Brăhmaņa, epics, etc. is flooded with information on animal care. The Purăņa are ancient scriptures discuss varied topics like devotion to God and his various aspects, traditional sciences like Ayurvĕda, Jyŏtişa (Astrology), cosmology, concepts like dharma, karma, reincarnation and many others. The treatment of animal diseases using Ayurvedic medicine has been mentioned in Garudapurăna, Agnipurăņa, Atri-samhită, Matsyapurăņa and many other texts. The Garudapurăņa is one of the important Săttvika purăna, the subject matter is divided into two parts, viz. Pŭrvakhaņda (first part) and an Uttarakhaņda (subsequent part). Gavăyurvĕda, Gajăyurvĕda narrated briefly and Aśvăyurvĕda described detailly in Pŭrvakhaņda.

  5. 32 CFR Appendix F to Part 623 - Power of Attorney (DA Form 4881-4-R)

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 3 2011-07-01 2009-07-01 true Power of Attorney (DA Form 4881-4-R) F Appendix F to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. F Appendix F to Part 623—Power of Attorney (DA Form...

  6. 32 CFR Appendix E to Part 623 - Surety Bond (DA Form 4881-3-R)

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 3 2010-07-01 2010-07-01 true Surety Bond (DA Form 4881-3-R) E Appendix E to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. E Appendix E to Part 623—Surety Bond (DA Form...

  7. 32 CFR Appendix E to Part 623 - Surety Bond (DA Form 4881-3-R)

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 3 2013-07-01 2013-07-01 false Surety Bond (DA Form 4881-3-R) E Appendix E to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. E Appendix E to Part 623—Surety Bond (DA Form 4881-3-R...

  8. 32 CFR Appendix E to Part 623 - Surety Bond (DA Form 4881-3-R)

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 3 2014-07-01 2014-07-01 false Surety Bond (DA Form 4881-3-R) E Appendix E to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. E Appendix E to Part 623—Surety Bond (DA Form 4881-3-R...

  9. 32 CFR Appendix E to Part 623 - Surety Bond (DA Form 4881-3-R)

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 3 2012-07-01 2009-07-01 true Surety Bond (DA Form 4881-3-R) E Appendix E to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. E Appendix E to Part 623—Surety Bond (DA Form 4881-3-R...

  10. Augmenting Effect of DA-9601 on Ghrelin in an Acute Gastric Injury Model

    PubMed Central

    Baek, Yoo Hum; Lee, Kang Nyeong; Jun, Dae Won; Yoon, Byung Chul; Kim, Ju Mi; Oh, Tae Young

    2011-01-01

    Background/Aims Acute gastric injury by alcohol or indomethacin has been reported to be prevented by DA-9601, an extract of the herb Artemisia asiatica. Ghrelin, an endogenously produced gastrointestinal peptide hormone, has also been demonstrated to play a role in gastric mucosal defense. The aim of this study was to investigate the effects of DA-9601 on ghrelin in an acute gastric injury model induced by alcohol or indomethacin. Methods A total of 140 Sprague-Dawley rats were divided into two groups, a placebo group and a DA-9601-pretreated group. Thirty minutes later, half of the rats in each group received ethanol injury and the other half received indomethacin injury. Levels of serum ghrelin and gastric mucosal ghrelin mRNA were measured by ELISA and RT-PCR, respectively. Results Immediately after ethanol administration, ghrelin increased in both groups pretreated with DA-9601 and placebo. However, the increase occurred more rapidly and was higher in the DA-9601-pretreated rats than in the controls that did not receive DA-9601-pretreatment. Similarly, from 30 minutes to 2 hours after indomethacin administration, the DA-9601-pretreated rats showed a significant increase in serum and gastric mucosal ghrelin concentrations, whereas placebo-pretreated rats showed only a mild increase. Conclusions DA-9601 potentiates the endogenous production and secretion of ghrelin in acute gastric injury models induced by ethanol or indomethacin. PMID:21461072

  11. A Creative Approach to the Common Core Standards: The Da Vinci Curriculum

    ERIC Educational Resources Information Center

    Chaucer, Harry

    2012-01-01

    "A Creative Approach to the Common Core Standards: The Da Vinci Curriculum" challenges educators to design programs that boldly embrace the Common Core State Standards by imaginatively drawing from the genius of great men and women such as Leonardo da Vinci. A central figure in the High Renaissance, Leonardo made extraordinary contributions as a…

  12. A Proposal to Build Evaluation Capacity at the Bunche-Da Vinci Learning Partnership Academy

    ERIC Educational Resources Information Center

    King, Jean A.

    2005-01-01

    The author describes potential evaluation capacity-building activities in contrast to the specifics of an evaluation design. Her response to the case of the Bunche-Da Vinci Learning Partnership Academy is developed in three parts: (1) an initial framing of the Bunche-Da Vinci situation; (2) what should be done before signing a contract; and (3)…

  13. A Creative Approach to the Common Core Standards: The Da Vinci Curriculum

    ERIC Educational Resources Information Center

    Chaucer, Harry

    2012-01-01

    "A Creative Approach to the Common Core Standards: The Da Vinci Curriculum" challenges educators to design programs that boldly embrace the Common Core State Standards by imaginatively drawing from the genius of great men and women such as Leonardo da Vinci. A central figure in the High Renaissance, Leonardo made extraordinary contributions as a…

  14. 32 CFR Appendix E to Part 623 - Surety Bond (DA Form 4881-3-R)

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 3 2011-07-01 2009-07-01 true Surety Bond (DA Form 4881-3-R) E Appendix E to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. E Appendix E to Part 623—Surety Bond (DA Form 4881-3-R...

  15. The analysis Arabidopsis thaliana overexpressing a 14kDa self-folding protein [abstract

    USDA-ARS?s Scientific Manuscript database

    A recent study in banana identified a 14kDa protein that has been hypothesized to function in regulating the nucleation and growth of the needle-shaped crystals of calcium oxalate that accumulate within the tissues of this plant. To gain further insight in to the functional role of this 14 kDa prote...

  16. The Indicative and Subjunctive "da"-complements in Serbian A Syntactic-Semantic Approach

    ERIC Educational Resources Information Center

    Todorovic, Natasa

    2012-01-01

    A syntactic-semantic investigation of subjunctive and indicative "da"-complements in Serbian is conducted in this project. After a careful comparison of Serbian sentence constructions with "da"-complements to the equivalent sentence structures in languages of the Balkans as well as other Slavic languages, it is clearly…

  17. Tyrosine phosphorylation of two cytosolic proteins of 50 kDa and 35 kDa in rat liver by insulin-receptor kinase in vitro.

    PubMed Central

    Kwok, Y C; Yip, C C

    1987-01-01

    Insulin-receptor tyrosine kinase can phosphorylate a variety of artificial substrates in vitro. Its physiological substrate(s), however, remains unknown. In the present study, we show that immobilized insulin receptors phosphorylate tyrosine residues of two cytosolic proteins of 50 kDa and 35 kDa in rat liver. Phosphorylation of these two proteins required Mn2+- or Mg2+-ATP as the phosphate donor. Phosphorylation was time- and temperature-dependent. Furthermore, the rate of phosphorylation of the two proteins was related to the autophosphorylated state of the insulin receptor. The pI of the phosphorylated 50 kDa and 35 kDa proteins was 5.4 and 5.6 respectively. These proteins were present in low abundance. They were not related to each other, nor to the insulin receptor, as demonstrated by in-gel proteolytic digestion and by immunoprecipitation using antibodies produced against them. They were specific substrates for the insulin receptor kinase, since they were not phosphorylated by epidermal-growth-factor-receptor kinase. These observations suggest that the 50 kDa and 35 kDa cytosolic proteins may be endogenous substrates for the insulin-receptor kinase. Images Fig. 1. Fig. 2. Fig. 5. Fig. 6. Fig. 7. Fig. 8. Fig. 9. PMID:2829823

  18. DA-9701: A New Multi-Acting Drug for the Treatment of Functional Dyspepsia.

    PubMed

    Kwon, Yong Sam; Son, Miwon

    2013-05-30

    Motilitone(®) (DA-9701) is a new herbal drug that was launched for the treatment of functional dyspepsia in December 2011 in Korea. The heterogeneous symptom pattern and multiple causes of functional dyspepsia have resulted in multiple drug target strategies for its treatment. DA-9701, a compound consisting of a combination of Corydalis Tuber and Pharbitidis Semen, has being developed for treatment of functional dyspepsia. It has multiple mechanisms of action such as fundus relaxation, visceral analgesia, and prokinetic effects. Furthermore, it was found to significantly enhance meal-induced gastric accommodation and increase gastric compliance in dogs. DA-9701 also showed an analgesic effect in rats with colorectal distension induced visceral hypersensitivity and an antinociceptive effect in beagle dogs with gastric distension-induced nociception. The pharmacological effects of DA-9701 also include conventional effects, such as enhanced gastric emptying and gastrointestinal transit. The safety profi le of DA-9701 is also preferable to that of other treatments.

  19. DA-9701: A New Multi-Acting Drug for the Treatment of Functional Dyspepsia

    PubMed Central

    Kwon, Yong Sam; Son, Miwon

    2013-01-01

    Motilitone® (DA-9701) is a new herbal drug that was launched for the treatment of functional dyspepsia in December 2011 in Korea. The heterogeneous symptom pattern and multiple causes of functional dyspepsia have resulted in multiple drug target strategies for its treatment. DA-9701, a compound consisting of a combination of Corydalis Tuber and Pharbitidis Semen, has being developed for treatment of functional dyspepsia. It has multiple mechanisms of action such as fundus relaxation, visceral analgesia, and prokinetic effects. Furthermore, it was found to significantly enhance meal-induced gastric accommodation and increase gastric compliance in dogs. DA-9701 also showed an analgesic effect in rats with colorectal distension induced visceral hypersensitivity and an antinociceptive effect in beagle dogs with gastric distension-induced nociception. The pharmacological effects of DA-9701 also include conventional effects, such as enhanced gastric emptying and gastrointestinal transit. The safety profi le of DA-9701 is also preferable to that of other treatments. PMID:24265862

  20. The 10 kDa protein of Taenia solium metacestodes shows genus specific antigenicity.

    PubMed

    Park, S K; Yun, D H; Chung, J Y; Kong, Y; Cho, S Y

    2000-09-01

    Genus specific antigenicity of the 10 kDa protein in cyst fluid (CF) of Taenia solium metacestodes was demonstrated by comparative immunoblot analysis. When CFs from taeniid metacestodes of T. saginata, T. solium, T. taeniaeformis and T. crassiceps were probed with specific monoclonal antibody (mAb) raised against 150 kDa protein of T. solium metacestodes, specific antibody reactions were observed in 7 and 10 kDa proteins of T. solium and in 7/8 kDa of T. saginata, T. taeniaeformis and T. crassiceps. The mAb did not react with any protein in hydatid fluid of Echinococcus granulosus and E. multilocularis. This result revealed that the 10 kDa peptide of T. solium metacestodes and its equivalent proteins of different Taenia metacestodes are genus specific antigens that are shared among different Taenia species.

  1. Growth-Promoting Hormone DA-6 Assists Phytoextraction and Detoxification of Cd by Ryegrass.

    PubMed

    He, Shanying; Wu, Qiuling; He, Zhenli

    2015-01-01

    A pot experiment was carried out to study the effect of growth-promoting hormone diethyl aminoethyl hexanoate (DA-6) on Cd phytoextraction and detoxification in ryegrass. Foliar spray of DA-6 significantly enhanced Cd extraction efficiency (P<0.05), with 1 μM DA-6 the most effective. At the subcellular level, 43-53% of Cd was soluble fraction and 23-46% in cell wall, and 9-25% in organelles. Chemical speciation analysis showed that 52.7-58.5% of Cd was NaCl extractable, 12.1-22.7% ethanol extractable, followed by other fractions. DA-6 alleviated metal toxicity by fixing more Cd in cell wall and decreasing Cd migration in plant. In conclusion, ryegrass tolerates Cd by cell wall compartmentalization along with protein and organic acids combination, and the treatment of 1 μM DA-6 appears to be optimal for enhancing the remediation efficiency of ryegrass for Cd contaminated soil.

  2. Xanthelasma and lipoma in Leonardo da Vinci's Mona Lisa.

    PubMed

    Dequeker, Jan; Muls, Erik; Leenders, Kathleen

    2004-08-01

    The painting Mona Lisa in the Louvre, Paris, by Leonardo da Vinci (1503-1506), shows skin alterations at the inner end of the left upper eyelid similar to xanthelasma, and a swelling of the dorsum of the right hand suggestive of a subcutaneous lipoma. These findings in a 25-30 year old woman, who died at the age of 37, may be indicative of essential hyperlipidemia, a strong risk factor for ischemic heart disease in middle age. As far as is known, this portrait of Mona Lisa painted in 1506 is the first evidence that xanthelasma and lipoma were prevalent in the sixteenth century, long before the first description by Addison and Gall in 1851.

  3. Leonardo da Vinci: the search for the soul.

    PubMed

    Del Maestro, R F

    1998-11-01

    The human race has always contemplated the question of the anatomical location of the soul. During the Renaissance the controversy crystallized into those individuals who supported the heart ("cardiocentric soul") and others who supported the brain ("cephalocentric soul") as the abode for this elusive entity. Leonardo da Vinci (1452-1519) joined a long list of other explorers in the "search for the soul." The method he used to resolve this anatomical problem involved the accumulation of information from ancient and contemporary sources, careful notetaking, discussions with acknowledged experts, and his own personal search for the truth. Leonardo used a myriad of innovative methods acquired from his knowledge of painting, sculpture, and architecture to define more clearly the site of the "senso comune"--the soul. In this review the author examines the sources of this ancient question, the knowledge base tapped by Leonardo for his personal search for the soul, and the views of key individuals who followed him.

  4. [Regarding the Manuscript D " Dell' occhio " of Leonardo da Vinci].

    PubMed

    Heitz, Robert F

    2009-01-01

    Leonardo da Vinci's Manuscript D consists of five double pages sheets, which, folded in two, comprise ten folios. This document, in the old Tuscan dialect and mirror writing, reveals the ideas of Leonardo on the anatomy of the eye in relation to the formation of images and visual perception. Leonardo explains in particular the behavior of the rays in the eye in terms of refraction and reflection, and is very mechanistic in his conception of the eye and of the visual process. The most significant innovations found in these folios are the concept of the eye as a camera obscura and the intersection of light rays in the interior of the eye. His texts nevertheless show hesitation, doubts and a troubled confusion, reflecting the ideas and uncertainties of his era. He did not share his results in his lifetime, despite both printing and etching being readily available to him.

  5. Sine ars scientia nihil est: Leonardo da Vinci and beyond.

    PubMed

    Kickhöfel, Eduardo H P

    2009-01-01

    The aim of this article is to reflect on the relationship between art and science so far as it concerns a symposium on neurosciences. We undertake a historical overview of that relationship, paying particular attention to the sui generis case of Leonardo da Vinci, who very often is regarded as the man who worked on art and science with equal ease. We then explain why his idea of merging these two forms of knowledge failed, considering the clear-cut distinction between art and science in his time. With this clarification, we explore the matter today. We look at Raphael's The Transfiguration, in which the representation of the possessed boy is seen by neuroscientists as indicative of an epileptic seizure. We also look at the ideas of neuroscientists Semir Zeki and Vilayanur Ramachandran, who study particular aspects of brain function and suggest a new merging of art and science.

  6. A 92-kDa human immunostimulatory protein.

    PubMed Central

    Fontan, E; Briend, E; Saklani-Jusforgues, H; d'Alayer, J; Vandekerckhove, J; Fauve, R M

    1994-01-01

    We purified to apparent homogeneity a human urinary glycoprotein of 92 kDa (HGP.92) that, administered intravenously at 250 micrograms/kg, fully protected mice against a lethal inoculum of Listeria monocytogenes. Since HGP.92 protected scid mice, which lack B and T lymphocytes, this increased resistance to Listeria did not appear to be lymphocyte mediated. Furthermore, inflammatory macrophages incubated with 6 nM HGP.92 inhibited the growth of Lewis carcinoma cells in vitro. These two activities appeared to depend on an oligosaccharide moiety, as they were lost after N-Glycanase treatment of HGP.92. Thus, the biological activity of HGP.92 was in some way related to a glycan moiety. Images PMID:8078887

  7. VLBI Radar of the 2012 DA14 Asteroid

    NASA Astrophysics Data System (ADS)

    Nechaeva, M. B.; Dugin, N. A.; Antipenko, A. A.; Bezrukov, D. A.; Bezrukov, V. V.; Voytyuk, V. V.; Dement'ev, A. F.; Jekabsons, N.; Klapers, M.; Konovalenko, A. A.; Kulishenko, V. F.; Nabatov, A. S.; Nesteruk, V. N.; Putillo, D.; Reznichenko, A. M.; Salerno, E.; Snegirev, S. D.; Tikhomirov, Yu. V.; Khutornoy, R. V.; Skirmante, K.; Shmeld, I.; Chagunin, A. K.

    2015-03-01

    An experiment on VLBI radar of the 2012 DA14 asteroid was carried out on February 15-16, 2011 at the time of its closest approach to the Earth. The research teams of Kharkov (Institute of Radio Astronomy of the National Academy of Sciences of Ukraine), Evpatoria (National Space Facilities Control and Test Center), Nizhny Novgorod (Radiophysical Research Institute), Bologna (Istituto di Radioastronomia (INAF)), and Ventspils (Ventspils International Radioastronomy Center) took part in the experiment. The asteroid was irradiated by the RT-70 planetary radar (Evpatoria) at a frequency of 5 GHz. The reflected signal was received using two 32-m radio telescopes in Medicina (Italy) and Irbene (Latvia) in radiointerferometric mode. The Doppler frequency shifts in bi-static radar mode and interference frequency in VLBI mode were measured. Accuracy of the VLBI radar method for determining the radial and angular velocities of the asteroid were estimated.

  8. Discovery of Photospheric Germanium in Hot DA White Dwarfs

    NASA Astrophysics Data System (ADS)

    Vennes, Stéphane; Chayer, Pierre; Dupuis, Jean

    2005-04-01

    We report the identification of Ge IV resonance lines in ultraviolet spectra of the hot DA white dwarfs Feige 24, G191-B2B, and GD 246. The lines originate in the stellar photosphere, and we measure low Ge/H abundance ratios ranging between -8.0 and -8.7. We also tentatively identify a resonance line of Sn IV blended with an Fe V line in the spectrum of G191-B2B. The presence of germanium extends our knowledge of the abundance pattern in hot white dwarfs beyond the iron group. The abundance ratio appears nearly solar, which implies either that the germanium abundance mixture in these stars has remained unaltered since leaving the main sequence or that diffusion processes (e.g., selective radiation pressure) are coincidentally reproducing a solar Ge/H ratio.

  9. 70 DA WHITE DWARFS IDENTIFIED IN LAMOST PILOT SURVEY

    SciTech Connect

    Zhao, J. K.; Luo, A. L.; Zhao, G.; Oswalt, T. D. E-mail: gzhao@bao.ac.cn E-mail: toswalt@fit.edu

    2013-06-01

    We present a spectroscopically identified catalog of 70 DA white dwarfs (WDs) from the LAMOST pilot survey. Thirty-five are found to be new identifications after cross-correlation with the Eisenstein et al. and Villanova catalogs. The effective temperature and gravity of these WDs are estimated by Balmer lines fitting. Most of them are hot WDs. The cooling times and masses of these WDs are estimated by interpolation in theoretical evolution tracks. The peak of the mass distribution is found to be {approx}0.6 M {sub Sun }, which is consistent with prior work in the literature. The distances of these WDs are estimated using the method of synthetic spectral distances. All of these WDs are found to be in the Galactic disk from our analysis of space motions. Our sample supports the expectation that WDs with high mass are concentrated near the plane of the Galactic disk.

  10. The First Six Outbursting Cool DA White Dwarf Pulsators

    NASA Astrophysics Data System (ADS)

    Bell, K. J.; Hermes, J. J.; Montgomery, M. H.; Winget, D. E.; Gentile Fusillo, N. P.; Raddi, R.; Gänsicke, B. T.

    2017-03-01

    Extensive observations from the Kepler spacecraft have recently revealed a new outburst phenomenon operating in cool pulsating DA (hydrogen atmosphere) white dwarfs (DAVs). With the introduction of two new outbursting DAVs from K2 Fields 7 (EPIC 229228364) and 8 (EPIC 220453225) in these proceedings, we presently know of six total members of this class of object. We present the observational commonalities of the outbursting DAVs: (1) outbursts that increase the mean stellar flux by up to ≍15%, last many hours, and recur irregularly on timescales of days; (2) effective temperatures that locate them near the cool edge of the DAV instability strip; and (3) rich pulsation spectra with modes that are observed to wander in amplitude/frequency.

  11. Trichinella spiralis: modulation of experimental autoimmune encephalomyelitis in DA rats.

    PubMed

    Gruden-Movsesijan, A; Ilic, N; Mostarica-Stojkovic, M; Stosic-Grujicic, S; Milic, M; Sofronic-Milosavljevic, Lj

    2008-04-01

    Helminth infection has a potent systemic immunomodulatory effect on the host immune response, which also affects the development of autoimmune diseases. We investigated the dose-dependent influence of Trichinella spiralis infection on experimental autoimmune encephalomyelitis (EAE). Our model of concomitant T. spiralis infection and EAE demonstrates that established infection of Dark Agouti (DA) rats with the parasite causes amelioration of the clinical course of induced EAE in a dose-dependent way. Infection with T. spiralis L1 stage muscle larvae (TSL1) reduced the severity of the autoimmune disease as judged by lower maximal clinical score, cumulative index, duration of illness and degree of mononuclear cell infiltration in T. spiralis infected animals compared to control, EAE-induced group. This study provides a valuable model of worm infection to investigate helminth-induced regulatory mechanisms for optimal benefit to the host.

  12. The DA{Phi}NE beam position monitors

    SciTech Connect

    Ghigo, A.; Sannibale, F.; Serio, M.; Vaccarezza, C.

    1997-01-01

    The beam diagnostics network of DA{Phi}NE, the Frascati {Phi}-factory, includes more than 110 beam position monitors divided between button monitors and striplines. The shape of the vacuum chamber changes along the accelerator implying several different geometries for these monitors. Moreover, in the two interaction regions of the collider where the electron and positron beams pass into the same chamber, a six-button configuration has been used. A bench calibration of each family of BPMs and striplines is being performed. A polynomial correction function has been derived by fitting the calibration results. An analytical-numerical analysis of the buttons` geometry has been done in order to compare the experimental with the theoretical results. {copyright} {ital 1997 American Institute of Physics.}

  13. Heavy-element abundance patterns in hot DA white dwarfs

    NASA Astrophysics Data System (ADS)

    Barstow, M. A.; Good, S. A.; Holberg, J. B.; Hubeny, I.; Bannister, N. P.; Bruhweiler, F. C.; Burleigh, M. R.; Napiwotzki, R.

    2003-05-01

    We present a series of systematic abundance measurements for 25 hot DA white dwarfs in the temperature range ~20 000-110 000 K, based on far-ultraviolet spectroscopy with the Space Telescope Imaging Spectrograph (STIS)/Goddard High Resolution Spectrograph (GHRS) on-board Hubble Space Telescope, IUE and FUSE. Using our latest heavy-element blanketed non-local thermodynamic equilibrium (non-LTE) stellar atmosphere calculations we have addressed the heavy-element abundance patterns, making completely objective measurements of abundance values and upper limits using a χ2 fitting technique to determine the uncertainties in the abundance measurements, which can be related to the formal upper limits in those stars where particular elements are not detected. We find that the presence or absence of heavy elements in the hot DA white dwarfs largely reflects what would be expected if radiative levitation is the supporting mechanism, although the measured abundances do not match the predicted values very well, as reported by other authors in the past. Almost all stars hotter than ~50 000 K contain heavy elements. For most of these the spread in element abundances is quite narrow and similar to the abundances measured in G191-B2B. However, there is an unexplained dichotomy at lower temperatures with some stars having apparently pure H envelopes and others having detectable quantities of heavy elements. The heavy elements present in these cooler stars are often stratified, lying in the outermost layers of the envelope. A few strong temperature/evolutionary effects are seen in the abundance measurements. There is a decreasing Si abundance with temperature, the N abundance pattern splits into two groups at lower temperature and there is a sharp decline in Fe and Ni abundance to zero, below ~50 000 K. When detected, the Fe and Ni abundances maintain an approximately constant ratio, close to the cosmic value of ~20. For the hottest white dwarfs observed by STIS, the strongest

  14. DaVinci-assisted laparoscopic radical prostatectomy: the learning curve

    NASA Astrophysics Data System (ADS)

    Le, Carter Q.; Ho, Khai-Linh V.; Gettman, Matthew T.

    2007-02-01

    Objective: To define the learning curve for daVinci-assisted laparoscopic radical prostatectomy (DLP) at our institution. Methods: The data from 170 patients who underwent DLP between August 2002 and December 2004 by a single surgeon (MTG) were reviewed. Operative time, hemoglobin decrease, conversion to open procedure, positive margin rates, complications, length of stay (LOS), length of catheterization, continence, and erectile function were analyzed. Results: Hemoglobin decrease (p=0.11), positive margin rates (p=0.80), and early urinary continence (p=0.17) did not significantly correlate with surgical experience. A trend towards lower complications (p=0.07) and an earlier return of erectile function (p=0.09) was noted with increased experience with DLP. Operative time, hospital stay, catheterization time, and open conversion showed significant association with patient sequence. Median operative time for the first 60 and the last 110 patients was 323.5 and 239.5 minutes (p=<0.0001), respectively. Median LOS for the aforementioned groups was 53 and 51 hours (p=0.009). Length of catheterization declined significantly between the first 60 and the remaining 110 patients, 14 as compared to 11.5 days (p=<0.0001). Eight open conversions occurred, six were in the first 30 patients (p=0.03). Conclusion: As an indicator of the learning curve, the operative time in our series showed no correlation with sequence after the 60 th patient. Thus, despite the advantages of robotics, the learning curve to efficient performance of daVinciassisted laparoscopic radical prostatectomy is long. Oncological and functional outcomes should not be affected during the learning curve.

  15. Therapeutic effect of DA-9601 on chronic reflux gastritis induced by sodium taurocholate in rats

    PubMed Central

    Oh, Tae Young; Shin, Chang Yell; Sohn, Yong Sung; Kim, Dong Hwan; Ahn, Byoung Ok; Lee, Eun Bang; Park, Cho Hyun

    2005-01-01

    AIM: To investigate the therapeutic effects of DA-9601 on sodium taurocholate (TCA)-induced chronic reflux gastritis in SD rats. METHODS: In this study, we have investigated the therapeutic effects of DA-9601 on chronic erosive and atrophic gastritis induced by 6 mo of TCA administration (5 mmol/L in drinking water) in SD rats. RESULTS: Four weeks of DA-9601 administration (0.065%, 0.216% in rat chow), following the withdrawal of TCA treatment, resulted in a significant decrease in total length of erosions in rats in a dose-dependent manner. Furthermore, the indicators of atrophic gastritis, such as reduced mucosal thickness and reduction in the number of parietal cells, were improved by the administration of DA-9601 in a dose-related manner. DA-9601 also attenuated inflammatory cell infiltration and the proliferation of collagenous fiber in the gastric mucosa. The improvement in the reduction of the gastric mucus was observed in the rats receiving a high dose of DA-9601 (0.216%). The therapeutic effect of DA-9601 on experimental chronic erosive gastritis was superior to that of rebamipide (1.08% in rat chow). Biochemical analyses showed increased mucosal prostaglandin E2 and reduced glutathione levels by DA-9601 treatment. CONCLUSION: We suggest that DA-9601 is a promising agent for the treatment of chronic erosive and atrophic gastritis with an etiological factor of bile reflux. Increased mucosal prostaglandin E2 and reduced glutathione by DA-9601 treatment may be therapeutic mechanisms for chronic erosive and atrophic gastritis. PMID:16437712

  16. Beyond the Classic VTA: Extended Amygdala Projections to DA-Striatal Paths in the Primate.

    PubMed

    Fudge, Julie L; Kelly, Emily A; Pal, Ria; Bedont, Joseph L; Park, Lydia; Ho, Brian

    2017-03-22

    The central extended amygdala (CEA) has been conceptualized as a 'macrosystem' that regulates various stress-induced behaviors. Consistent with this, the CEA highly expresses corticotropin-releasing factor (CRF), an important modulator of stress responses. Stress alters goal-directed responses associated with striatal paths, including maladaptive responses such as drug seeking, social withdrawal, and compulsive behavior. CEA inputs to the midbrain dopamine (DA) system are positioned to influence striatal functions through mesolimbic DA-striatal pathways. However, the structure of this amygdala-CEA-DA neuron path to the striatum has been poorly characterized in primates. In primates, we combined neuronal tracer injections into various arms of the circuit through specific DA subpopulations to assess: (1) whether the circuit connecting amygdala, CEA, and DA cells follows CEA intrinsic organization, or a more direct topography involving bed nucleus vs central nucleus divisions; (2) CRF content of the CEA-DA path; and (3) striatal subregions specifically involved in CEA-DA-striatal loops. We found that the amygdala-CEA-DA path follows macrostructural subdivisions, with the majority of input/outputs converging in the medial central nucleus, the sublenticular extended amygdala, and the posterior lateral bed nucleus of the stria terminalis. The proportion of CRF+ outputs is >50%, and mainly targets the A10 parabrachial pigmented nucleus (PBP) and A8 (retrorubal field, RRF) neuronal subpopulations, with additional inputs to the dorsal A9 neurons. CRF-enriched CEA-DA projections are positioned to influence outputs to the 'limbic-associative' striatum, which is distinct from striatal regions targeted by DA cells lacking CEA input. We conclude that the concept of the CEA is supported on connectional grounds, and that CEA termination over the PBP and RRF neuronal populations can influence striatal circuits involved in associative learning.Neuropsychopharmacology advance online

  17. Pharmacological action of DA-9701 on the motility of feline stomach circular smooth muscle.

    PubMed

    Nguyen, Thanh Thao; Song, Hyun Ju; Ko, Sung Kwon; Sohn, Uy Dong

    2015-03-01

    DA-9701, a new prokinetic agent for the treatment of functional dyspepsia, is formulated with Pharbitis semen and Corydalis tuber. This study wasconducted to determine the pharmacological action of DA-9701 and to identify the receptors involved in DA-9701 -induced contractile responsesin the feline gastric corporal, fundic and antral circular smooth muscle. Concentration-response curve to DA-9701 was established. The tissue trips were exposed to methylsergide, ketanserin, ondansetron, GR 113808, atropine and dopamine before administration of DA-9701. The contractile force was determined before and after administration of drugs by a polygraph.DA-9701 enhanced the spontaneous contractile amplitude of antrum, corpus and fundus. However, it did not change the spontaneous contractile frequency of antrum and corpus, but concentration-dependently reduced that of fundus. In the fundus, DA-9701 -induced tonic contractions were inhibited by dopamine, methylsergide, ketanserine, ondansetron or GR 113808 respectively, but not by atropine, indicating that the contractile responses are mediated by multiple receptors: 5-HT2, 5-HT3, 5-HT4, and dopamine receptors. In the corpus, DA-9701-induced contractions were blocked by atropine, dopamine or GR 113808, but not by methysergide, ketanserin or ondansetron, indicating that they are involved in receptors on both, smooth muscles and neurons: 5-HT4 and dopamine receptors. However, contractile responses to DA-9701 are mainly mediated by dopamine receptors in the antrum. These results suggest that DA-9701 has important roles in gastric accommodation by enhancing tonic activity of fundus, and in gastric emptying and gastrointestinal transit by phasic contractions of corpus and antrum mediated by multiple receptors.

  18. Reduced striatal dopamine DA D2 receptor function in dominant-negative GSK-3 transgenic mice.

    PubMed

    Gomez-Sintes, Raquel; Bortolozzi, Analia; Artigas, Francesc; Lucas, José J

    2014-09-01

    Glycogen synthase kinase-3 (GSK-3) is a serine/threonine kinase with constitutive activity involved in cellular architecture, gene expression, cell proliferation, fate decision and apoptosis, among others. GSK-3 expression is particularly high in brain where it may be involved in neurological and psychiatric disorders such as Alzheimer׳s disease, bipolar disorder and major depression. A link with schizophrenia is suggested by the antipsychotic drug-induced GSK-3 regulation and by the involvement of the Akt/GSK-3 pathway in dopaminergic neurotransmission. Taking advantage of the previous development of dominant negative GSK-3 transgenic mice (Tg) showing a selective reduction of GSK-3 activity in forebrain neurons but not in dopaminergic neurons, we explored the relationship between GSK-3 and dopaminergic neurotransmission in vivo. In microdialysis experiments, local quinpirole (DA D2-R agonist) in dorsal striatum reduced dopamine (DA) release significantly less in Tg mice than in wild-type (WT) mice. However, local SKF-81297 (selective DA D1-R agonist) in dorsal striatum reduced DA release equally in both control and Tg mice indicating a comparable function of DA D1-R in the direct striato-nigral pathway. Likewise, systemic quinpirole administration - acting preferentially on presynaptic DA D2- autoreceptors to modulate DA release-reduced striatal DA release similarly in both control and Tg mice. Quinpirole reduced locomotor activity and induced c-fos expression in globus pallidus (both striatal DA D2-R-mediated effects) significantly more in WT than in Tg mice. Taking together, the present results show that dominant negative GSK-3 transgenic mice show reduced DA D2-R-mediated function in striatum and further support a link between dopaminergic neurotransmission and GSK-3 activity. Copyright © 2014 Elsevier B.V. and ECNP. All rights reserved.

  19. The Effect of DA-9701 in Opioid-induced Bowel Dysfunction of Guinea Pig

    PubMed Central

    Hussain, Zahid; Rhee, Kwang Won; Lee, Young Ju; Park, Hyojin

    2016-01-01

    Background/Aims Opioid induced bowel dysfunction (OIBD) is associated with decreased gastrointestinal (GI) propulsive activity due to intake of opioid analgesics. DA-9701, a novel prokinetic agent formulated with Pharbitis Semen and Corydalis Tuber has promising effects on GI motor function. Therefore, we aim to evaluate the prokinetic effects of DA-9701 in an OIBD model of guinea pig. Methods The ileal and distal colon muscle contraction in presence of different doses of DA-9701, morphine, and combination (morphine + DA-9701) was measured by tissue bath study. The prokinetic effect of DA-9701 was assessed by charcoal transit and fecal pellet output assay in an OIBD model of guinea pig. Results DA-9701 significantly increased the amplitude and area under the curve of ileal muscle contraction, while there was insignificant effect on the distal colon compared to the control. The maximal amplitude of ileal muscle contraction was acquired at a concentration of 10 μg/mL of DA-9701. In contrast, morphine significantly decreased the amplitude of ileal and distal colon muscle contraction compared to the control. Morphine delayed both upper (P < 0.01) and lower (P < 0.05) GI transit, and delayed GI transit was restored by the administration of DA-9701. Morphine induced reduction of contractility was significantly ameliorated by addition of DA-9701 in both ileal and distal colon muscles. Conclusions DA-9701 significantly increased the amplitude of contraction of the ileal muscle, however the distal colon muscle contraction was insignificant. Additionally, it restored delayed upper and lower GI transit in an OIBD model of guinea pig, and it might prove to be a useful candidate drug in a clinical trial for OIBD. PMID:26932898

  20. Beyond Donor-Acceptor (D-A) Approach: Structure-Optoelectronic Properties-Organic Photovoltaic Performance Correlation in New D-A1 -D-A2 Low-Bandgap Conjugated Polymers.

    PubMed

    Chochos, Christos L; Drakopoulou, Sofia; Katsouras, Athanasios; Squeo, Benedetta M; Sprau, Christian; Colsmann, Alexander; Gregoriou, Vasilis G; Cando, Alex-Palma; Allard, Sybille; Scherf, Ullrich; Gasparini, Nicola; Kazerouni, Negar; Ameri, Tayebeh; Brabec, Christoph J; Avgeropoulos, Apostolos

    2017-04-01

    Low-bandgap near-infrared polymers are usually synthesized using the common donor-acceptor (D-A) approach. However, recently polymer chemists are introducing more complex chemical concepts for better fine tuning of their optoelectronic properties. Usually these studies are limited to one or two polymer examples in each case study so far, though. In this study, the dependence of optoelectronic and macroscopic (device performance) properties in a series of six new D-A1 -D-A2 low bandgap semiconducting polymers is reported for the first time. Correlation between the chemical structure of single-component polymer films and their optoelectronic properties has been achieved in terms of absorption maxima, optical bandgap, ionization potential, and electron affinity. Preliminary organic photovoltaic results based on blends of the D-A1 -D-A2 polymers as the electron donor mixed with the fullerene derivative [6,6]-phenyl-C71 -butyric acid methyl ester demonstrate power conversion efficiencies close to 4% with short-circuit current densities (J sc ) of around 11 mA cm(-2) , high fill factors up to 0.70, and high open-circuit voltages (V oc s) of 0.70 V. All the devices are fabricated in an inverted architecture with the photoactive layer processed in air with doctor blade technique, showing the compatibility with roll-to-roll large-scale manufacturing processes.

  1. 43 kDa and 66 kDa, two blood stage antigens induce immune response in Plasmodium berghei malaria.

    PubMed

    Pirta, Chhaya; Banyal, H S

    2014-08-01

    The hunt for an effective vaccine against malaria still continues. Several new target antigens as candidates for vaccine design are being explored and tested for their efficacy. In the present study the sera from mice immunized with 24,000 x g fraction of Plasmodium berghei has been used to identify highly immunogenic blood stage antigens. The protective antibodies present in immune sera were covalently immobilized on CNBr activated sepharose 4B and used for affinity chromatography purification of antigens present in blood stages of P. berghei. Two polypeptides of 66 and 43 kDa molecular weights proved to be highly immunogenic. They exhibited a strong humoral immune response in mice as evident by high titres in ELISA and IFA. Protective immunity by these two antigens was apparent by in vivo and in vitro studies. These two proteins could further be analysed and used as antigens in malaria vaccine design.

  2. Essential roles of Da transactivation domains in neurogenesis and in E(spl)-mediated repression.

    PubMed

    Zarifi, Ioanna; Kiparaki, Marianthi; Koumbanakis, Konstantinos A; Giagtzoglou, Nikolaos; Zacharioudaki, Evanthia; Alexiadis, Anastasios; Livadaras, Ioannis; Delidakis, Christos

    2012-11-01

    E proteins are a special class of basic helix-loop-helix (bHLH) proteins that heterodimerize with many bHLH activators to regulate developmental decisions, such as myogenesis and neurogenesis. Daughterless (Da) is the sole E protein in Drosophila and is ubiquitously expressed. We have characterized two transcription activation domains (TADs) in Da, called activation domain 1 (AD1) and loop-helix (LH), and have evaluated their roles in promoting peripheral neurogenesis. In this context, Da heterodimerizes with proneural proteins, such as Scute (Sc), which is dynamically expressed and also contributes a TAD. We found that either one of the Da TADs in the Da/Sc complex is sufficient to promote neurogenesis, whereas the Sc TAD is incapable of doing so. Besides its transcriptional activation role, the Da AD1 domain serves as an interaction platform for E(spl) proteins, bHLH-Orange family repressors which antagonize Da/Sc function. We show that the E(spl) Orange domain is needed for this interaction and strongly contributes to the antiproneural activity of E(spl) proteins. We present a mechanistic model on the interplay of these bHLH factors in the context of neural fate assignment.

  3. Biofortification of soybean meal: immunological properties of the 27 kDa γ-zein.

    PubMed

    Krishnan, Hari B; Jang, Sungchan; Kim, Won-Seok; Kerley, Monty S; Oliver, Melvin J; Trick, Harold N

    2011-02-23

    Legumes, including soybeans ( Glycine max ), are deficient in sulfur-containing amino acids, which are required for the optimal growth of monogastric animals. This deficiency can be overcome by expressing heterologous proteins rich in sulfur-containing amino acids in soybean seeds. A maize 27 kDa γ-zein, a cysteine-rich protein, has been successfully expressed in several crops including soybean, barley, and alfalfa with the intent to biofortify these crops for animal feed. Previous work has shown that the maize 27 kDa zein can withstand digestion by pepsin and elicit an immunogenic response in young pigs. By use of sera from patients who tested positive by ImmunoCAP assay for elevated IgE to maize proteins, specific IgE binding to the 27 kDa γ-zein is demonstrated. Bioinformatic analysis using the full-length and 80 amino acid sliding window FASTA searches identified significant sequence homology of the 27 kDa γ-zein with several known allergens. Immunoblot analysis using human serum that cross-reacts with maize seed proteins also revealed specific IgE-binding to the 27 kDa γ-zein in soybean seed protein extracts containing the 27 kDa zein. This study demonstrates for the first time the allergenicity potential of the 27 kDa γ-zein and the potential that this protein has to limit livestock performance when used in soybeans that serve as a biofortified feed supplement.

  4. Endogenous dopamine (DA) modulates (3H)spiperone binding in vivo in rat brain

    SciTech Connect

    Bischoff, S.; Krauss, J.; Grunenwald, C.; Gunst, F.; Heinrich, M.; Schaub, M.; Stoecklin, K.V.; Vassout, A.; Waldmeier, P.; Maitre, L. )

    1991-01-01

    (3H)spiperone (SPI) binding in vivo, biochemical parameters and behavior were measured after modulating DA levels by various drug treatments. DA releasers and uptake inhibitors increased SPI binding in rat striatum. In other brain areas, the effects were variable, but only the pituitary remained unaffected. Surprisingly, nomifensine decreased SPI binding in frontal cortex. The effects of these drugs were monitored by measuring DA, serotonin (5-HT) and their metabolites in the same rats. The increased SPI binding in striatum was parallel to the locomotor stimulation with the following rank order: amfonelic acid greater than nomifensine greater than D-amphetamine greater than or equal to methylphenidate greater than amineptine greater than bupropion. Decreasing DA levels with reserpine or alpha-methyl-para-tyrosine reduced SPI binding by 45% in striatum only when both drugs were combined. In contrast, reserpine enhanced SPI binding in pituitary. Thus, the amount of releasable DA seems to modulate SPI binding characteristics. It is suggested that in vivo, DA receptors are submitted to dynamic regulation in response to changes in intrasynaptic concentrations of DA.

  5. Vectra DA for the objective measurement of disease activity in patients with rheumatoid arthritis.

    PubMed

    Segurado, O G; Sasso, E H

    2014-01-01

    Quantitative and regular assessment of disease activity in rheumatoid arthritis (RA) is required to achieve treatment targets such as remission and to optimize clinical outcomes. To assess inflammation accurately, predict joint damage and monitor treatment response, a measure of disease activity in RA should reflect the pathological processes resulting in irreversible joint damage and functional disability. The Vectra DA blood test is an objective measure of disease activity for patients with RA. Vectra DA provides an accurate, reproducible score on a scale of 1 to 100 based on the concentrations of 12 biomarkers that reflect the pathophysiologic diversity of RA. The analytical validity, clinical validity, and clinical utility of Vectra DA have been evaluated for patients with RA in registries and prospective and retrospective clinical studies. As a biomarker-based instrument for assessing disease activity in RA, the Vectra DA test can help monitor therapeutic response to methotrexate and biologic agents and assess clinically challenging situations, such as when clinical measures are confounded by non-inflammatory pain from fibromyalgia. Vectra DA scores correlate with imaging of joint inflammation and are predictive for radiographic progression, with high Vectra DA scores being associated with more frequent and severe progression and low scores being predictive for non-progression. In summary, the Vectra DA score is an objective measure of RA disease activity that quantifies inflammatory status. By predicting risk for joint damage more effectively than conventional clinical and laboratory measures, it has the potential to complement these measures and optimise clinical decision making.

  6. The ductus arteriosus migratory smooth muscle cell phenotype processes tropoelastin to a 52-kDa product associated with impaired assembly of elastic laminae.

    PubMed

    Hinek, A; Rabinovitch, M

    1993-01-15

    We established the identity of a 52-kDa protein secreted by fetal lamb ductus arteriosus (DA) smooth muscle cells (SMC) and suggest how it might be related to structural changes unique to DA development, i.e. reduced assembly of elastic laminae and associated formation of intimal cushions. We produced a monoclonal antibody (HI-20) to the 52-kDa protein and observed, by electron microscopy, immunogold labeling of elastin in both DA and aorta vessel walls. Western immunoblotting showed that HI-20, as well as antibodies to tropoelastin, reacted with the 52-kDa protein secreted by DA SMC, as well as with 68-kDa tropoelastin. The highly specific antibody to the carboxyl-terminal sequence of tropoelastin failed, however, to recognize the 52-kDa protein, although it reacted well with the 68-kDa tropoelastin. Amino acid analysis and sequencing data confirmed the identity of the affinity-purified 52-kDa protein as truncated tropoelastin with an intact amino terminus. Cell-free translation of mRNA extracted from DA and aorta SMC produced a 68-kDa, but not a 52-kDa, immunoprecipitated tropoelastin. When DA and aorta SMC were pulsed with [14C]valine, we immunoprecipitated, after only a 15-min chase, both 68-kDa and 52-kDa tropoelastin from cell extracts of DA SMC, but only the 68-kDa tropoelastin was present in aorta SMC. There was no evidence of proteolytic degradation of radiolabeled aorta 68-kDa tropoelastin to a 52-kDa species when mixed with DA SMC conditioned medium. This suggests that the 52-kDa tropoelastin is the result of cell-associated processing or degradation of an original 68-kDa product of translation. Furthermore, pulse-chase experiments showed initial secretion of equivalent amounts of 68-kDa and 52-kDa tropoelastins by cultured DA SMC with increasing accumulation of the 52-kDa species, suggesting its impaired insolubilization. The production, in high concentration, of a 52-kDa tropoelastin product that lacks the carboxyl terminus, may prevent its alignment

  7. Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

    PubMed Central

    Wei, Hung-Mu; Lin, Li-Chih; Wang, Chiu-Feng; Lee, Yi-Jang; Chen, Yuan-Tsong; Liao, You-Di

    2016-01-01

    Granulysin, a cationic protein expressed by human natural killer cells and cytotoxic T lymphocytes, is a mediator for drug-induced Stevens-Johnson syndrome and graft-versus-host disease. Some 15 kDa granulysin are processed into 9 kDa forms and sequestered in cytolytic granules, while others are constitutively secreted into body fluids. Both 9 and 15 kDa granulysin have been shown to be a serum marker for cell-mediated immunity. Furthermore, 15 kDa is able to activate monocyte differentiation. However, its antimicrobial properties have not been clearly addressed. Here, we report a novel method to prepare both the soluble 9 and 15 kDa granulysin and show that the 15 kDa form is more effective than the 9 kDa form in exerting specific antimicrobial activity against Pseudomonas aeruginosa within a range of few micromolars. We also show that the 15 kDa granulysin is able to hyperpolarize the membrane potential and increase membrane permeability of treated bacteria. Interestingly, the bactericidal activity and membrane permeability of the granulysins were markedly reduced at lower pH (pH 5.4) as a result of probable increase in hydrophobicity of the granulysins. Additionally, we’ve also shown the granulysin to inhibit biofilm formation by P. aeruginosa. These results suggest that the 15 kDa granulysin exhibits a novel mechanism in bacteria killing in a way that’s different from most antimicrobial peptides. Our novel granulysin preparation methodology will be useful for further study of action mechanisms of other antimicrobial, cytotoxic and immunomodulating properties in granulysin-mediated diseases. PMID:27276051

  8. Therapeutic effect of infused Fluosol-DA/carbogen with ephedrine, flunarizine, or nitroprusside

    SciTech Connect

    Teicher, B.A.; Holden, S.A.; Northey, D.; Dewhirst, M.W.; Herman, T.S.

    1993-04-30

    The perfluorochemical emulsion Fluosol-DA plus carbogen breathing has been shown to increase the effectiveness of radiation therapy in preclinical solid tumors when the emulsion was administered by i.v. bolus injection. Much of the enhancement in tumor radiation response was lost when the emulsion was administered slowly. The authors hypothesized that an increase in tumor perfusion resulted when Fluosol-DA was administered rapidly. In the present study, the [alpha]/[beta] agonist ephedrine, the Ca[sup 2+] channel blocker flunarizine and the nitric oxide producing vasodilating drug nitroprusside have been tested. Ephedrine administration resulted in a decrease in the radiation plus Fluosol-DA [+-] carbogen antitumor effects in both the Lewis lung carcinoma and FSaIIC tumor systems. In contrast, flunarizine administration resulted in an increase in the efficacy of the radiation plus carbogen and the radiation plus Fluosol-DA/carbogen in both emulsion was given rapidly. Even with flunarizine administration Fluosol-DA delivered slowly was less effective than when the emulsion was given rapidly. Flunarizine with Fluosol-DA infused i.v. over 30 min followed by carbogen breathing prior to and during radiation therapy resulted in a 1.7-1.6-fold increase in response compared with 2.4-2.2-fold with Fluosol-DA administered by injection i.v. and carbogen breathing prior to and during radiation therapy using growth delay of the Lewis lung carcinoma. The effects of nitroprusside were complex. This drug had considerably more effect at 10 Gy than at higher radiation doses. These studies suggest that Fluosol-DA given by i.v. injection may increase tumor perfusion and that a drug like flunarizine may be beneficial if the Fluosol-DA is administered slowly followed by carbogen breathing and radiation therapy. 45 refs., 4 figs.

  9. Pharmacokinetics of intravenous and oral DA-8159, a new erectogenic, in rats with protein-calorie malnutrition.

    PubMed

    Shim, Hyun J; Kim, Yu C; Lee, Joo H; Ahn, Byung O; Kwon, Jong W; Kim, Won B; Lee, Inchul; Lee, Myung G

    2004-12-01

    Influence of dietary protein deficiency on the pharmacokinetics of DA-8159 and one of its metabolites, DA-8164, was investigated after intravenous and oral administration of DA-8159 at a dose of 30 mg kg(-1) to male Sprague-Dawley rats allowed free access to a 23% (control) or 5% (protein-calorie malnutrition, PCM) casein diet for 4 weeks. The total area under the plasma concentration-time curve from time zero to time infinity (AUC) values of DA-8164 were significantly smaller after both intravenous (87.0 vs 162 microg min mL(-1)) and oral (144 vs 319 microg min mL(-1)) administration of DA-8159 to PCM rats. This could be due to the decrease in CYP3A1/2 (50-60%) in the rats because DA-8164 was mainly formed via CYP3A1/2 in rats. This could be supported by significantly slower in-vitro CL(int) (2.04+/-0.646 vs 3.15+/-0.693 microL min(-1) (mg protein)(-1)) for the formation of DA-8164 in hepatic microsomal fraction of PCM rats. After intravenous administration of DA-8159, the AUC values of DA-8159 were not significantly different between the two groups of rats although the AUC of DA-8164 was significantly smaller in PCM rats, and this may be due to the minor metabolic pathway of DA-8164 in rats. However, after oral administration of DA-8159, the AUC of DA-8159 was significantly greater in PCM rats (194 vs 122 microg min mL(-1)). This was not due to enhanced absorption of DA-8159 from the gastrointestinal tract in the rats but may be due to a decreased intestinal first-pass effect of DA-8159 in the rats.

  10. Use of the new da Vinci Xi® during robotic rectal resection for cancer: a pilot matched-case comparison with the da Vinci Si®.

    PubMed

    Morelli, Luca; Guadagni, Simone; Di Franco, Gregorio; Palmeri, Matteo; Caprili, Giovanni; D'Isidoro, Cristiano; Cobuccio, Luigi; Marciano, Emanuele; Di Candio, Giulio; Mosca, Franco

    2017-03-01

    The aim of this study was to compare the short-term outcomes of robotic rectal resection with total mesorectal excision (TME) for rectal cancer, with the use of the new da Vinci Xi® (Xi-RobTME group) and the da Vinci Si® (Si-RobTME group). Ten patients with histologically confirmed rectal cancer underwent robot-assisted TME with the use of the new da Vinci Xi. The outcomes of Xi-RobTME group were compared with a Si-RobTME group selected using a case-matched methodology. Overall operative times and mean hospital stays were shorter in the Xi-RobTME group. Surgeries were fully robotic with a complete take-down of the splenic flexure in all Xi-RobTME cases, while only four cases of the Si-RobTME group were fully robotic, with two cases of complete take-down of the splenic flexure. The new da Vinci Xi could offer some advantages with respect to the da Vinci Si in rectal resection for cancer. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  11. MALDI-TOF High Mass Calibration up to 200 kDa Using Human Recombinant 16 kDa Protein Histidine Phosphatase Aggregates

    PubMed Central

    Ludwig, Katrin; Habbach, Schähdi; Krieglstein, Josef; Klumpp, Susanne; König, Simone

    2011-01-01

    Background Protein histidine phosphatase (PHP) is an enzyme which removes phosphate groups from histidine residues. It was described for vertebrates in the year 2002. The recombinant human 16 kDa protein forms multimeric complexes in physiological buffer and in the gas phase. High-mass calibration in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has remained a problem due to the lack of suitable standards. Large proteins can hardly be freed of their substructural microheterogeneity by classical purification procedures so that their use as calibrants is limited. A small adduct-forming protein of validated quality is a valuable alternative for that purpose. Methodology/Principal Findings Three major PHP clusters of ∼113, 209 and >600 kDa were observed in gel filtration analysis. Re-chromatography of the monomer peak showed the same cluster distribution. The tendency to associate was detected also in MALDI-TOF MS measuring regular adducts up to 200 kDa. Conclusions/Significance PHP forms multimers consisting of up to more than 35 protein molecules. In MALDI-TOF MS it generates adduct ions every 16 kDa. The protein can be produced with high quality so that its use as calibration compound for high mass ranges above 100 kDa, where standards are difficult to obtain, is feasible. PMID:21876758

  12. Respostas religiosas à aids no Brasil: impressões de pesquisa acerca da Pastoral de DST/Aids da Igreja Católica1

    PubMed Central

    Seffner, Fernando; Silva, Cristiane Gonçalves Meireles da; Maksud, Ívia; Garcia, Jonathan; Rios, Luís Felipe; Natividade, Marcelo; Borges, Priscila Rodrigues; Parker, Richard; Terto, Veriano

    2009-01-01

    O texto encontra-se estruturado em quatro partes. Na primeira delas, apresentamos um conjunto de considerações e informações acerca da situação da aids no Brasil, das relações entre religião, sexualidade, aids e estado laico, bem como uma descrição mais clara do Projeto Respostas Religiosas ao HIV/Aids no Brasil, do qual este texto apresenta algumas impressões de pesquisa preliminares. A seguir, dedicamos um item a apresentação da Pastoral de DST/Aids, sua história, estrutura e objetivos. No item seguinte problematizamos diversas questões em particular no âmbito das relações Estado e Igreja, relações entre agentes de pastoral e hierarquia da Igreja, e questões ligadas mais diretamente à sexualidade e aids, todas referenciadas ao trabalho da Pastoral de DST/Aids. Ao final, apresentamos a bibliografia e fontes consultadas. PMID:20442806

  13. Myelin management by the 18.5-kDa and 21.5-kDa classic myelin basic protein isoforms.

    PubMed

    Harauz, George; Boggs, Joan M

    2013-05-01

    The classic myelin basic protein (MBP) splice isoforms range in nominal molecular mass from 14 to 21.5 kDa, and arise from the gene in the oligodendrocyte lineage (Golli) in maturing oligodendrocytes. The 18.5-kDa isoform that predominates in adult myelin adheres the cytosolic surfaces of oligodendrocyte membranes together, and forms a two-dimensional molecular sieve restricting protein diffusion into compact myelin. However, this protein has additional roles including cytoskeletal assembly and membrane extension, binding to SH3-domains, participation in Fyn-mediated signaling pathways, sequestration of phosphoinositides, and maintenance of calcium homeostasis. Of the diverse post-translational modifications of this isoform, phosphorylation is the most dynamic, and modulates 18.5-kDa MBP's protein-membrane and protein-protein interactions, indicative of a rich repertoire of functions. In developing and mature myelin, phosphorylation can result in microdomain or even nuclear targeting of the protein, supporting the conclusion that 18.5-kDa MBP has significant roles beyond membrane adhesion. The full-length, early-developmental 21.5-kDa splice isoform is predominantly karyophilic due to a non-traditional P-Y nuclear localization signal, with effects such as promotion of oligodendrocyte proliferation. We discuss in vitro and recent in vivo evidence for multifunctionality of these classic basic proteins of myelin, and argue for a systematic evaluation of the temporal and spatial distributions of these protein isoforms, and their modified variants, during oligodendrocyte differentiation.

  14. 2012 DA14 and Chelyabinsk: Same Day Coincidence?

    NASA Astrophysics Data System (ADS)

    Chapman, Clark R.

    2013-10-01

    A long anticipated near-miss by near-Earth asteroid (NEA) 2012 DA14 on 15 February 2013 was upstaged 16 hours earlier by the impact and atmospheric explosion of a ~20 m NEA over Chelyabinsk, Russia. DA14 was earlier estimated to be 45 m across and passage at a distance under geosynchronous satellites was considered to be a record close approach by an object of that size, a once-in-40-years event. Actual Earth impact by a 20 m NEA had been estimated to be a once-in-200-years event. A simplistic calculation gives a probability of these happening on the same day of 1-in-a-billion. Within hours of the Chelyabinsk impact, it was recognized and widely reported that the two asteroids were in very different orbits and could not, at least in any readily understandable way, be physically related to each other (e.g. fragments of the same precursor body). Also, some early reports suggested that the two asteroids had very different compositions, further undermining the possibility they were pieces of the same body. (It seems unlikely, though certainly conceivable, that the two NEAs could have different compositions yet have some kind of causal connection resulting in the same-day events.) Nevertheless, incredibly tiny probabilities beg for an explanation. Here, with the benefit of hindsight, I re-examine issues that affect the probabilities, such as size, albedo, probable composition, and numbers of NEAs of these sizes. I also consider difficult issues of framing the apparent coincidence, which dominate other uncertainties. Updated information about the physical nature of the two asteroids somewhat modifies the original estimates of probabilities. Moreover, more proper ways of framing elements of the coincidence considerably reduce the improbability of the same-day events, but not nearly to the level (e.g. 1-in-1000 or perhaps 1-in-10,000) where we could rationalize dismissing the events as “just a coincidence.” The events of 15 February 2013 deserve more sophisticated

  15. Leonardo Da Vinci and stroke - vegetarian diet as a possible cause.

    PubMed

    Oztürk, Serefnur; Altieri, Marta; Troisi, Pina

    2010-01-01

    Leonardo da Vinci (April 15, 1452 to May 2, 1519) was an Italian Renaissance architect, musician, anatomist, inventor, engineer, sculptor, geometer, and painter. It has been gleaned from the many available historical documents that da Vinci was a vegetarian who respected and loved animals, and that he suffered from right hemiparesis in the last 5 years of his life. A vegetarian diet has both positive and negative influences on the cerebrovascular system. In this report, a possible relation between a vegetarian diet and stroke is discussed from various perspectives as related to Leonardo da Vinci's stroke. Copyright (c) 2010 S. Karger AG, Basel.

  16. PD2P: PanDA Dynamic Data Placement for ATLAS

    SciTech Connect

    Maeno, T.; De, K.; Panitkin, S.

    2012-12-13

    The PanDA (Production and Distributed Analysis) system plays a key role in the ATLAS distributed computing infrastructure. PanDA is the ATLAS workload management system for processing all Monte-Carlo (MC) simulation and data reprocessing jobs in addition to user and group analysis jobs. The PanDA Dynamic Data Placement (PD2P) system has been developed to cope with difficulties of data placement for ATLAS. We will describe the design of the new system, its performance during the past year of data taking, dramatic improvements it has brought about in the efficient use of storage and processing resources, and plans for the future.

  17. DaVinci canvas: a telerobotic surgical system with integrated, robot-assisted, laparoscopic ultrasound capability.

    PubMed

    Leven, Joshua; Burschka, Darius; Kumar, Rajesh; Zhang, Gary; Blumenkranz, Steve; Dai, Xiangtian Donald; Awad, Mike; Hager, Gregory D; Marohn, Mike; Choti, Mike; Hasser, Chris; Taylor, Russell H

    2005-01-01

    We present daVinci Canvas: a telerobotic surgical system with integrated robot-assisted laparoscopic ultrasound capability. DaVinci Canvas consists of the integration of a rigid laparoscopic ultrasound probe with the daVinci robot, video tracking of ultrasound probe motions, endoscope and ultrasound calibration and registration, autonomous robot motions, and the display of registered 2D and 3D ultrasound images. Although we used laparoscopic liver cancer surgery as a focusing application, our broader aim was the development of a versatile system that would be useful for many procedures.

  18. PD2P: PanDA Dynamic Data Placement for ATLAS

    NASA Astrophysics Data System (ADS)

    Maeno, T.; De, K.; Panitkin, S.

    2012-12-01

    The PanDA (Production and Distributed Analysis) system plays a key role in the ATLAS distributed computing infrastructure. PanDA is the ATLAS workload management system for processing all Monte-Carlo (MC) simulation and data reprocessing jobs in addition to user and group analysis jobs. The PanDA Dynamic Data Placement (PD2P) system has been developed to cope with difficulties of data placement for ATLAS. We will describe the design of the new system, its performance during the past year of data taking, dramatic improvements it has brought about in the efficient use of storage and processing resources, and plans for the future.

  19. Arbitrary Amplitude DIA and DA Solitary Waves in Adiabatic Dusty Plasmas

    SciTech Connect

    Mamun, A. A.; Jahan, N.; Shukla, P. K.

    2008-10-15

    The dust-ion-acoustic (DIA) as well as the dust-acoustic (DA) solitary waves (SWs) in an adiabatic dusty plasma are investigated by the pseudo-potential approach which is valid for arbitrary amplitude SWs. The role of the adiabaticity of electrons and ions in modifying the basic features (polarity, speed, amplitude and width) of arbitrary amplitude DIA and DA SWs are explicitly examined. It is found that the effects of the adiabaticity of electrons and ions significantly modify the basic features (polarity, speed, amplitude and width) of the DIA and DA SWs. The implications of our results in space and laboratory dusty plasmas are briefly discussed.

  20. Evolving understanding of translocator protein 18 kDa (TSPO).

    PubMed

    Li, Fei; Liu, Jian; Garavito, R Michael; Ferguson-Miller, Shelagh

    2015-09-01

    The translocator protein 18 kDa (TSPO) has been the focus of intense research by the biomedical community and the pharmaceutical industry because of its apparent involvement in many disease-related processes. These include steroidogenesis, apoptosis, inflammation, neurological disease and cancer, resulting in the use of TSPO as a biomarker and its potential as a drug target. Despite more than 30 years of study, the precise function of TSPO remains elusive. A recent breakthrough in determining the high-resolution crystal structures of bacterial homologs of mitochondrial TSPO provides new insight into the structural and functional properties at a molecular level and new opportunities for investigating the significance of this ancient and highly conserved protein family. The availability of atomic level structural information from different species also provides a platform for structure-based drug development. Here we briefly review current knowledge regarding TSPO and the implications of the new structures with respect to hypotheses and controversies in the field. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Robotic Partial Nephrectomy with the Da Vinci Xi.

    PubMed

    Kallingal, George J S; Swain, Sanjaya; Darwiche, Fadi; Punnen, Sanoj; Manoharan, Murugesan; Gonzalgo, Mark L; Parekh, Dipen J

    2016-01-01

    Purpose. The surgical expertise to perform robotic partial nephrectomy is heavily dependent on technology. The Da Vinci Xi (XI) is the latest robotic surgical platform with significant advancements compared to its predecessor. We describe our operative technique and experience with the XI system for robotic partial nephrectomy (RPN). Materials and Methods. Patients with clinical T1 renal masses were offered RPN with the XI. We used laser targeting, autopositioning, and a novel "in-line" port placement to perform RPN. Results. 15 patients underwent RPN with the XI. There were no intraoperative complications and no operative conversions. Mean console time was 101.3 minutes (range 44-176 minutes). Mean ischemia time was 17.5 minutes and estimated blood loss was 120 mLs. 12 of 15 patients had renal cell carcinoma. Two patients had oncocytoma and one had benign cystic disease. All patients had negative surgical margins and pathologic T1 disease. Two postoperative complications were encountered, including one patient who developed a pseudoaneurysm and one readmitted for presumed urinary tract infection. Conclusions. RPN with the XI system can be safely performed. Combining our surgical technique with the technological advancements on the XI offers patients acceptable pathologic and perioperative outcomes.

  2. Leonardo da Vinci: engineer, bioengineer, anatomist, and artist.

    PubMed

    West, John B

    2017-03-01

    Leonardo da Vinci (1452-1519) enjoys a reputation as one of the most talented people of all time in the history of science and the arts. However, little attention has been given to his contributions to physiology. One of his main interests was engineering, and he was fascinated by structural problems and the flow patterns of liquids. He also produced a large number of ingenious designs for warfare and a variety of highly original flying machines. But of particular interest to us are his contributions to bioengineering and how he used his knowledge of basic physical principles to throw light on physiological function. For example, he produced new insights into the mechanics of breathing including the action of the ribs and diaphragm. He was the first person to understand the different roles of the internal and external intercostal muscles. He had novel ideas about the airways including the mode of airflow in them. He also worked on the cardiovascular system and had a special interest in the pulmonary circulation. But, interestingly, he was not able to completely divorce his views from those of Galen, in that although he could not see pores in the interventricular septum of the heart, one of his drawings included them. Leonardo was a talented anatomist who made many striking drawings of the human body. Finally, his reputation for many people is based on his paintings including the Mona Lisa that apparently attracts more viewers than any other painting in the world.

  3. [Project Leonardo-da-Vinci for better nursing care].

    PubMed

    Gábor, Katalin; Csanádi, Lajosné; Helembai, Kornélia; Szögi, Zoltánné; Tulkán, Ibolya; Unginé, Kántor Katalin

    2002-08-18

    The aim of the present paper is to inform physicians about the work completed by nurses and professors of baccalaureat nurses in the framework of Leonardo da Vinci project, organised and sponsored by the European Union. The goal of the project was to increase the effectiveness of chief nurses throughout their further training programme in the field of management. The team of Szeged chose the human resource management, since in this field is possible to achieve the greatest improvement with the smallest financial investment. We measured the fluctuations and the absentees of the nurses, the changes in the degree of education, the nurse' and patient' satisfaction at the beginning and at the end of the period studied. Except the patient's satisfaction all the other parameters improved by the end of tested period. The project provided a unique possibility to compare the state of the Hungarian nursing with that of the countries belonging to the European Union, to exchange the experience and to learn some new methods. In the framework of this project a book of two volumes was prepared containing the suggestions of EU. This book is widely available in English and in French.

  4. New Middle Pleistocene hominin cranium from Gruta da Aroeira (Portugal)

    PubMed Central

    Daura, Joan; Sanz, Montserrat; Arsuaga, Juan Luis; Hoffmann, Dirk L.; Quam, Rolf M.; Ortega, María Cruz; Santos, Elena; Gómez, Sandra; Rubio, Angel; Villaescusa, Lucía; Souto, Pedro; Mauricio, João; Rodrigues, Filipa; Ferreira, Artur; Godinho, Paulo; Trinkaus, Erik

    2017-01-01

    The Middle Pleistocene is a crucial time period for studying human evolution in Europe, because it marks the appearance of both fossil hominins ancestral to the later Neandertals and the Acheulean technology. Nevertheless, European sites containing well-dated human remains associated with an Acheulean toolkit remain scarce. The earliest European hominin crania associated with Acheulean handaxes are at the sites of Arago, Atapuerca Sima de los Huesos (SH), and Swanscombe, dating to 400–500 ka (Marine Isotope Stage 11–12). The Atapuerca (SH) fossils and the Swanscombe cranium belong to the Neandertal clade, whereas the Arago hominins have been attributed to an incipient stage of Neandertal evolution, to Homo heidelbergensis, or to a subspecies of Homo erectus. A recently discovered cranium (Aroeira 3) from the Gruta da Aroeira (Almonda karst system, Portugal) dating to 390–436 ka provides important evidence on the earliest European Acheulean-bearing hominins. This cranium is represented by most of the right half of a calvarium (with the exception of the missing occipital bone) and a fragmentary right maxilla preserving part of the nasal floor and two fragmentary molars. The combination of traits in the Aroeira 3 cranium augments the previously documented diversity in the European Middle Pleistocene fossil record. PMID:28289213

  5. New Middle Pleistocene hominin cranium from Gruta da Aroeira (Portugal).

    PubMed

    Daura, Joan; Sanz, Montserrat; Arsuaga, Juan Luis; Hoffmann, Dirk L; Quam, Rolf M; Ortega, María Cruz; Santos, Elena; Gómez, Sandra; Rubio, Angel; Villaescusa, Lucía; Souto, Pedro; Mauricio, João; Rodrigues, Filipa; Ferreira, Artur; Godinho, Paulo; Trinkaus, Erik; Zilhão, João

    2017-03-28

    The Middle Pleistocene is a crucial time period for studying human evolution in Europe, because it marks the appearance of both fossil hominins ancestral to the later Neandertals and the Acheulean technology. Nevertheless, European sites containing well-dated human remains associated with an Acheulean toolkit remain scarce. The earliest European hominin crania associated with Acheulean handaxes are at the sites of Arago, Atapuerca Sima de los Huesos (SH), and Swanscombe, dating to 400-500 ka (Marine Isotope Stage 11-12). The Atapuerca (SH) fossils and the Swanscombe cranium belong to the Neandertal clade, whereas the Arago hominins have been attributed to an incipient stage of Neandertal evolution, to Homo heidelbergensis, or to a subspecies of Homo erectus A recently discovered cranium (Aroeira 3) from the Gruta da Aroeira (Almonda karst system, Portugal) dating to 390-436 ka provides important evidence on the earliest European Acheulean-bearing hominins. This cranium is represented by most of the right half of a calvarium (with the exception of the missing occipital bone) and a fragmentary right maxilla preserving part of the nasal floor and two fragmentary molars. The combination of traits in the Aroeira 3 cranium augments the previously documented diversity in the European Middle Pleistocene fossil record.

  6. DA WHITE DWARFS OBSERVED IN THE LAMOST PILOT SURVEY

    SciTech Connect

    Zhang Yueyang; Deng Licai; Liu Chao; Carrell, Kenneth; Yang Fan; Gao Shuang; Xu Yan; Li Jing; Zhang Haotong; Zhao Yongheng; Luo Ali; Bai Zhongrui; Yuan Hailong; Lepine, Sebastien; Newberg, Heidi Jo; Carlin, Jeffrey L.; Jin Ge

    2013-08-01

    A total of {approx}640, 000 objects from the LAMOST pilot survey have been publicly released. In this work, we present a catalog of DA white dwarfs (DAWDs) from the entire pilot survey. We outline a new algorithm for the selection of white dwarfs (WDs) by fitting Sersic profiles to the Balmer H{beta}, H{gamma}, and H{delta} lines of the spectra, and calculating the equivalent width of the Ca II K line. Two thousand nine hundred sixty-four candidates are selected by constraining the fitting parameters and the equivalent width of the Ca II K line. All the spectra of candidates are visually inspected. We identify 230 DAWDs (59 of which are already included in the Villanova and SDSS WD catalogs), 20 of which are DAWDs with non-degenerate companions. In addition, 128 candidates are classified as DAWDs/subdwarfs, which means the classifications are ambiguous. The result is consistent with the expected DAWD number estimated based on the LEGUE target selection algorithm.

  7. Near-UV absorption in very cool DA white dwarfs

    SciTech Connect

    Saumon, D.; Holberg, J. B.; Kowalski, P. M. E-mail: holberg@argus.lpl.arizona.edu

    2014-07-20

    The atmospheres of very cool, hydrogen-rich white dwarfs (WDs) (T{sub eff} < 6000 K) are challenging to model because of the increased complexity of the equation of state, chemical equilibrium, and opacity sources in a low-temperature, weakly ionized dense gas. In particular, many models that assume relatively simple models for the broadening of atomic levels and mostly ideal gas physics overestimate the flux in the blue part of their spectra. A solution to this problem that has met with some success is that additional opacity at short wavelengths comes for the extreme broadening of the Lyman α line of atomic H by collisions primarily with H{sub 2}. For the purpose of validating this model more rigorously, we acquired Hubble Space Telescope STIS spectra of eight very cool WDs (five DA and three DC stars). Combined with their known parallaxes, BVRIJHK, and Spitzer IRAC photometry, we analyze their entire spectral energy distribution (from 0.24 to 9.3 μm) with a large grid of model atmospheres and synthetic spectra. We find that the red wing of the Lyman α line reproduces the rapidly decreasing near-UV flux of these very cool stars very well. We determine better constrained values of T{sub eff} and gravity as well as upper limits to the helium abundance in their atmospheres.

  8. Biogeochemical Indicators of Paleoenvironmental Changes in a Humid Region of Amazonia (Lagoa da Pata, Sao Gabriel da Cachoeira)

    NASA Astrophysics Data System (ADS)

    Cordeiro, R. C.; Turcq, B.; Mendes, L. G.; Azeredo, J. B.; Moura, M. S.; de Oliveira, A. R.; Monteiro, F.; Santelli, R. E.

    2004-12-01

    Analysis of four sediment core collected in Lagoa da Pata lake show significant changes in the environmental history of Amazonia during the last 40,000 years. The Lagoa da Pata Lake (0°17'S, 66°40'W) is located on the top of the Morro dos Seis Lagos, an isolated hill in the extensive forested plane surface of high Rio Negro basin. The climate in the region presents mean annual precipitation around 3000 mm without dry season. Three distinct sections are clearly identified in the LPT III, LPT IV and LPT V cores. They consist of upper and lower organic-rich layers, separated by a clastic layer which represents a short period of rapid accumulation around 18 ka BP. Analysis of the organic matter composition by d13C and d15N, total Carbon and Nitrogen, sedimentary chlorophyll concentration and organic matter petrography show three different phases related to organic matter deposition. Between, at least 46,000 and 31,000 14C yr B.P., the lake presented a high water level attested by high accumulation rates of organic carbon. Values of d13C (around -32%) associated to high C/N ratio demonstrate a lignocellulosic material contribution in this phase; the second phase between the 31000 yr 14C B.P. and 18000 14C B.P. is characterised by a lower productivity attested by low sedimentary chlorophyll and total organic carbon values. The d13C values increase with medium values around -28% that may correspond to an input of organic matter rich in C4 plants (grasses). The sedimentation rate decreases significantly. High charcoal fluxes were observed in this period. At around 18000 14C yr BP, there appears to have been a sudden input of clastic material. This is represented by a sandy facies, which exhibits lower carbon contents. From 18000 14C yr B.P. to the present time occurred an increase in lacustrine productivity marked by an increase in carbon and chlorophyll derivate accumulation rates. The C/N values dropped indicting an algae organic matter contribution. It probably

  9. Novel morpholine scaffolds as selective dopamine (DA) D3 receptor antagonists.

    PubMed

    Micheli, Fabrizio; Cremonesi, Susanna; Semeraro, Teresa; Tarsi, Luca; Tomelleri, Silvia; Cavanni, Paolo; Oliosi, Beatrice; Perdonà, Elisabetta; Sava, Anna; Zonzini, Laura; Feriani, Aldo; Braggio, Simone; Heidbreder, Christian

    2016-02-15

    A new series of morpholine derivatives has been identified as selective DA D3 receptor antagonists; their in vitro profile and pharmacokinetic data are provided. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. The influence of ancient Greek thought on fifteenth century anatomy: Galenic influence and Leonardo da Vinci.

    PubMed

    Tubbs, Richard Isaiah; Gonzales, Jocelyn; Iwanaga, Joe; Loukas, Marios; Oskouian, Rod J; Tubbs, R Shane

    2017-05-29

    Leonardo da Vinci (1452-1519) can be called one of the earliest contributors to the history of anatomy and, by extension, the study of medicine. He may have even overshadowed Andreas Vesalius (1514-1564), the so-called founder of human anatomy, if his works had been published within his lifetime. While some of the best illustrations of their time, with our modern knowledge of anatomy, it is clear that many of da Vinci's depictions of human anatomy are inaccurate. However, he also made significant discoveries in anatomy and remarkable predictions of facts he could not yet discover with the technology available to him. Additionally, da Vinci was largely influenced by Greek anatomists, as indicated from his ideas about anatomical structure. In this historical review, we describe da Vinci's history, influences, and discoveries in anatomical research and his depictions and errors with regards to the musculoskeletal system, cardiovascular system, nervous system, and other organs.

  11. [RESEARCH PROGRESS OF PERIPHERAL NERVE SURGERY ASSISTED BY Da Vinci ROBOTIC SYSTEM].

    PubMed

    Shen, Jie; Song, Diyu; Wang, Xiaoyu; Wang, Changjiang; Zhang, Shuming

    2016-02-01

    To summarize the research progress of peripheral nerve surgery assisted by Da Vinci robotic system. The recent domestic and international articles about peripheral nerve surgery assisted by Da Vinci robotic system were reviewed and summarized. Compared with conventional microsurgery, peripheral nerve surgery assisted by Da Vinci robotic system has distinctive advantages, such as elimination of physiological tremors and three-dimensional high-resolution vision. It is possible to perform robot assisted limb nerve surgery using either the traditional brachial plexus approach or the mini-invasive approach. The development of Da Vinci robotic system has revealed new perspectives in peripheral nerve surgery. But it has still been at the initial stage, more basic and clinical researches are still needed.

  12. Confirmatory factor analysis of the Questionnaire on the Health Staff's Perceptions Regarding Doutores da Alegria's Interventions.

    PubMed

    Masetti, Morgana; Caires, Susana; Brandão, Daniel; Vieira, Diana Aguiar

    2016-06-09

    A confirmatory analysis was performed to validate the Questionnaire on the Health Staff's Perceptions Regarding Doutores da Alegria's Intervention, a 40-item instrument designed to assess pediatric staff's perceptions regarding the effects of Doutores da Alegria, a Brazilian hospital clowning professional organization. Eight dimensions were evaluated: the permanence of Doutores da Alegria's interventions; Doutores da Alegria's intrapersonal and interpersonal effects on their relation to health staff; themselves; staff-children; and staff-family relationships; as well as their effect on staff's cultural development; children's relation to their own disease; and families' attitude regarding their child's condition. In all, 567 health professionals from 13 Brazilian hospitals participated. The instrument's good psychometric features are acknowledged.

  13. DaG 1066: A Newfound Anomalous Ureilite with Chondritic Inclusions

    NASA Astrophysics Data System (ADS)

    Moggi Cecchi, V.; Caporali, S.; Pratesi, G.

    2015-07-01

    General description, textural, and compositional features of the polymict ureilite DaG 1066, recovered in 1999 in Libya, are provided. The meteorite contains various inclusions, among which almost pure forsterite and enstatite-bearing chondrules.

  14. Thinking like Leonardo da Vinci and its implications for the modern doctor.

    PubMed

    Baum, Neil

    2013-01-01

    Most people when asked to name the most creative, innovative, and multidimensional people in history would agree that Leonardo da Vinci is either at the top or very close to the number one position on that list. Wouldn't it be nice to think like da Vinci? This article shares the seven unique principles of thinking that da Vinci used that enabled him to be the greatest painter, sculptor, architect, musician, mathematician, engineer, inventor, anatomist, geologist, cartographer, botanist, and writer of his (if not of all) time. This article will take you deep into the notebooks and codices of da Vinci, and suggest ways his ideas can be used by anyone in the healthcare profession to make them a better healthcare provider.

  15. Temperature and Gravity Dependence of Trace Element Abundances in Hot DA White Dwarfs (94-EUVE-094)

    NASA Technical Reports Server (NTRS)

    Finley, David S.

    1998-01-01

    EUV spectroscopy has shown that DA white dwarfs hotter than about 45,000 K may contain trace heavy elements, while those hotter than about 50,000 K almost always have significant abundances of trace heavy elements. One of our continuing challenges is to identify and determine the abundances of these trace constituents, and then to relate the observed abundance patterns to the present conditions and previous evolutionary histories of the hot DA white dwarfs.

  16. Early clinical experience with the da Vinci Xi Surgical System in general surgery.

    PubMed

    Hagen, Monika E; Jung, Minoa K; Ris, Frederic; Fakhro, Jassim; Buchs, Nicolas C; Buehler, Leo; Morel, Philippe

    2016-12-27

    The da Vinci Xi Surgical System (Intuitive Surgical Inc., Sunnyvale, CA, USA) has been released in 2014 to facilitate minimally invasive surgery. Novel features are targeted towards facilitating complex multi-quadrant procedures, but data is scarce so far. Perioperative data of patients who underwent robotic general surgery with the da Vinci Xi system within the first 6 month after installation were collected and analyzed. The gastric bypass procedures performed with the da Vinci Xi Surgical System were compared to an equal amount of the last procedures with the da Vinci Si Surgical System. Thirty-one foregut (28 Roux-en-Y gastric bypasses), 6 colorectal procedures and 1 revisional biliary procedure were performed. The mean operating room (OR) time was 221.8 (±69.0) minutes for gastric bypasses and 306.5 (±48.8) for colorectal procedures with mean docking time of 9.4 (±3.8) minutes. The gastric bypass procedure was transitioned from a hybrid to a fully robotic approach. In comparison to the last 28 gastric bypass procedures performed with the da Vinci Si Surgical System, the OR time was comparable (226.9 versus 230.6 min, p = 0.8094), but the docking time significantly longer with the da Vinci Xi Surgical System (8.5 versus 6.1 min, p = 0.0415). All colorectal procedures were performed with a single robotic docking. No intraoperative and two postoperative complications occurred. The da Vinci Xi might facilitate single-setups of totally robotic gastric bypass and colorectal surgeries. However, further comparable research is needed to clearly determine the significance of this latest version of the da Vinci Surgical System.

  17. 32 CFR Appendix F to Part 623 - Power of Attorney (DA Form 4881-4-R)

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 3 2010-07-01 2010-07-01 true Power of Attorney (DA Form 4881-4-R) F Appendix F to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. F Appendix F to Part 623—Power of Attorney (DA Form 4881-4-R...

  18. Degradation of the COL1 domain of type XIV collagen by 92-kDa gelatinase.

    PubMed

    Sires, U I; Dublet, B; Aubert-Foucher, E; van der Rest, M; Welgus, H G

    1995-01-20

    Type XIV collagen is a newly described member of the fibril-associated collagens with interrupted triple helices (FACITs). Expression of this collagen has been localized to various embryonic tissues, suggesting that it has a functional role in development. All FACITs thus far described (types IX, XII, XIV, and XVI) contain a highly homologous carboxyl-terminal triple helical domain designated COL1. We have studied the capacity of various matrix metalloproteinases (interstitial collagenase, stromelysin, matrilysin, and 92-kDa gelatinase) to degrade the COL1 domain of collagen XIV. We found that only 92-kDa gelatinase cleaves COL1. Furthermore, digestion of whole native collagen XIV by the 92-kDa gelatinase indicates that this enzyme specifically attacks the carboxyl-terminal triple helix-containing region of the molecule. COL1 is cleaved by 92-kDa gelatinase at 30 degrees C, a full 5-6 degrees C below the melting temperature (Tm) of this domain; native collagen XIV is also degraded at 30 degrees C. In comparison to interstitial collagenase degradation of its physiologic native type I collagen substrate, the 92-kDa enzyme cleaved COL1 (XIV) with comparable catalytic efficacy. Interestingly, following thermal denaturation of the COL1 fragment, its susceptibility to 92-kDa gelatinase increases, but only to a degree that leaves it several orders of magnitude less sensitive to degradation than denatured collagens I and III. These data indicate that native COL1 and collagen XIV are readily and specifically cleaved by 92-kDa gelatinase. They also suggest a role for 92-kDa gelatinase activity in the structural tissue remodeling of the developing embryo.

  19. PAN-DA and beyond: Data acquisition for the next generation experiments

    SciTech Connect

    Pordes, R.; Anderson, J.; Berg, D.; Berman, E.; Brown, D.; Dorries, T.; Mackinnon, B.; Meadows, J.; Moore, C.; Nicinski, T.; Oleynik, G.; Petravick, D.; Rechenmacher, R.; Sergey, G.; Slimmer, D.; Streets, J.; Vittone, M.; Votava, M.; Wilcer, N.; White, V.

    1991-06-01

    We report on the status of the PAN-DA data acquisition system presented at the last Real Time Conference. Since that time, PAN-DA has been successfully used in the fixed target program at Fermilab. We also report on the plans and strategies for development of a new data acquisition system for the next generation of fixed target experiments at Fermilab. 10 refs., 3 figs.

  20. An Organizational Behavior View of Department of the Army (DA) Comptrollership.

    DTIC Science & Technology

    1981-12-01

    111-8 Number and Percentage of Comptroller Respondents from the Different DA MACOMs ------- 100 111-9 Summary of Descriptive Statistics for the...subsamples included 17 comptrollers from different DA MACOMs such as USAREUR, Material Development and Readiness Command (DARCOM), Forces Command (FORSCOM...organizations are configured as depicted in Figure 11-4. While the responsibilities of the division, installation and MACOM comptrollers are in many ways

  1. Load evaluation of the da Vinci surgical system for transoral robotic surgery.

    PubMed

    Fujiwara, Kazunori; Fukuhara, Takahiro; Niimi, Koji; Sato, Takahiro; Kitano, Hiroya

    2015-12-01

    Transoral robotic surgery, performed with the da Vinci surgical system (da Vinci), is a surgical approach for benign and malignant lesions of the oral cavity and laryngopharynx. It provides several unique advantages, which include a 3-dimensional magnified view and ability to see and work around curves or angles. However, the current da Vinci surgical system does not provide haptic feedback. This is problematic because the potential risks specific to the transoral use of the da Vinci include tooth injury, mucosal laceration, ocular injury and mandibular fracture. To assess the potential for intraoperative injuries, we measured the load of the endoscope and the instrument of the da Vinci Si surgical system. We pressed the endoscope and instrument of the da Vinci Si against Load cell six times each and measured the dynamic load and the time-to-maximum load. We also struck the da Vinci Si endoscope and instrument against the Load cell six times each and measured the impact load. The maximum dynamic load was 7.27 ± 1.31 kg for the endoscope and 1.90 ± 0.72 for the instrument. The corresponding time-to-maximum loads were 1.72 ± 0.22 and 1.29 ± 0.34 s, but the impact loads were significantly lower than the dynamic load. It remains possible that a major load is exerted on adjacent structures by continuous contact with the endoscope and instrument of da Vinci Si. However, there is a minor delay in reaching the maximum load. Careful monitoring by an on-site assistant may, therefore, help prevent contiguous injury.

  2. Diagnostic epitope variability within Taenia solium 8 kDa antigen family: implications for cysticercosis immunodetection.

    PubMed

    Ferrer, Elizabeth; Sánchez, Jipsy; Milano, Adriana; Alvarez, Suhei; La Rosa, Rosamelia; Lares, María; González, Luís Miguel; Cortéz, María Milagros; Dávila, Iris; Harrison, Leslie J S; Parkhouse, R Michael E; Gárate, Teresa

    2012-01-01

    To study diagnostic epitopes within the Taenia solium 8 kDa antigen family, six overlapping synthetic peptides from an 8 kDa family member (Ts8B2) were synthesized and evaluated by ELISA and MABA with sera from patients with neurocysticercosis (NCC), from infected pigs and from rabbits immunized with recombinant Ts8B2 protein. The pre-immune rabbit sera and the Ts8B2 recombinant protein served as negative and positive controls, respectively. A similar analysis was done with the already described antigenic peptides from another member of the 8 kDa family, highly similar to Ts8B2, the CyDA antigen. Surprisingly, neither the Ts8B2 peptides nor the CyDA peptides were recognized by infected human and porcine sera. However, the entire Ts8B2 recombinant, as well as amino and carboxy-terminal halves were recognized by the positive serum samples. The observed lack of recognition of linear Ts8B2 peptides suggests that the principal serological response to the Ts8B2 family is focused on conformational epitopes in contrast to the previously observed antigenicity of the CyDA peptides. This differential antigenicity of 8 kDa family peptides could be related with parasite antigenic variability. The fact that rabbits experimentally immunized with Ts8B2 did make anti-peptide antibodies to peptides Ts8B2-6 and CyDA-6, located in the carboxy-terminal region demonstrated that the Ts8B2 peptides are not intrinsically non-immunogenic.

  3. The sandfly fauna (Diptera: Psychodidae: Phlebotominae) of the Parque Estadual da Serra da Tiririca, Rio de Janeiro, Brazil.

    PubMed

    Rodrigues, Andressa Alencastre Fuzari; Barbosa, Vanessa de Araújo; Andrade Filho, José Dilermando; Brazil, Reginaldo Peçanha

    2013-11-01

    Cutaneous leishmaniasis (CL) in the state of Rio de Janeiro is sporadic and can be characterised as a peridomestic transmission that occurs in modified natural environments. The aim of this work was to study the fauna and ecological characteristics of sandflies in an environmentally protected area (the State Park of Serra da Tiririca) within the remnants of the Atlantic Forest in the municipalities of Niterói and Maricá and their possible relationship with leishmaniasis. Captures were performed using light traps during the night once a month for one year in both sylvatic environments and areas surrounding homes near the park. A total of 1,037 sandflies were captured, belonging to nine genera and 12 species: Evandromyia tupynambai (34.1%), Migonemyia migonei (20.6%), Brumptomyia cunhai (13.8%), Micropygomyia schreiberi (9.7%), Psathyromyia lanei (6.5%), Brumptomyia nitzulescui (5.7%), Evandromyia edwardsi (5.4%), Nyssomyia intermedia (2.8%), Evandromyia cortelezzii (0.6%), Pintomyia bianchigalatiae (0.5%), Lutzomyia longipalpis (0.2%) and Sciopemyia microps (0.1%). Both Mg. migonei and Ny. intermedia may be acting as vectors of CL in this area.

  4. The sandfly fauna (Diptera: Psychodidae: Phlebotominae) of the Parque Estadual da Serra da Tiririca, Rio de Janeiro, Brazil

    PubMed Central

    Rodrigues, Andressa Alencastre Fuzari; Barbosa, Vanessa de Araújo; Andrade, José Dilermando; Brazil, Reginaldo Peçanha

    2013-01-01

    Cutaneous leishmaniasis (CL) in the state of Rio de Janeiro is sporadic and can be characterised as a peridomestic transmission that occurs in modified natural environments. The aim of this work was to study the fauna and ecological characteristics of sandflies in an environmentally protected area (the State Park of Serra da Tiririca) within the remnants of the Atlantic Forest in the municipalities of Niterói and Maricá and their possible relationship with leishmaniasis. Captures were performed using light traps during the night once a month for one year in both sylvatic environments and areas surrounding homes near the park. A total of 1,037 sandflies were captured, belonging to nine genera and 12 species: Evandromyia tupynambai (34.1%), Migonemyia migonei (20.6%), Brumptomyia cunhai (13.8%), Micropygomyia schreiberi (9.7%), Psathyromyia lanei (6.5%), Brumptomyia nitzulescui (5.7%), Evandromyia edwardsi (5.4%), Nyssomyia intermedia (2.8%), Evandromyia cortelezzii (0.6%), Pintomyia bianchigalatiae (0.5%), Lutzomyia longipalpis (0.2%) and Sciopemyia microps (0.1%). Both Mg. migonei and Ny. intermedia may be acting as vectors of CL in this area. PMID:24141956

  5. DA 5505: a novel topical formulation of terbinafine that enhances skin penetration and retention.

    PubMed

    Thapa, Raj Kumar; Han, Sang-Duk; Park, Hyoung Geun; Son, Miwon; Jun, Joon Ho; Kim, Jong Oh

    2015-01-01

    Topical fungal infections can become severe if left untreated. Efficient treatment modalities for topical fungal infections aid the penetration of antifungal agents deep into viable skin layers. Terbinafine is a fungicidal agent that inhibits ergosterol, an essential fungal component. The main objective of this study was to evaluate skin permeation and retention of a terbinafine-loaded solution containing chitosan as a film former. Comparative assessment of skin permeation and retention was performed using a prepared formulation (DA 5505) and marketed formulations of terbinafine in murine and porcine skin. To mimic fungal infection of skin, keratinized skin was induced in NC/Nga mice. In comparison with the marketed formulations, DA 5505 exhibited significantly better skin permeation. The flux, permeation coefficient, and enhancement ratio of terbinafine were remarkably increased by DA 5505 in comparison with the marketed formulations, and lag time was dramatically reduced. DA 5505 significantly increased cumulative terbinafine retention in viable skin layers in comparison with the marketed solution, suggesting enhanced efficacy. Furthermore, DA 5505 exhibited superior skin permeation in normal skin and keratinized skin. Thus, the DA 5505 formulation has the potential to effectively deliver terbinafine to superficial and deep cutaneous fungal infections.

  6. Purification and partial characterization of a 31-kDa cysteine endopeptidase from germinated barley.

    PubMed

    Zhang, N; Jones, B L

    1996-01-01

    Proteolytic enzymes hydrolyze cereal seed storage proteins into small peptides and amino acids, which are very important for seed germination and the malting process. A cysteine-class endopeptidase was purified from 4-d-germinated barley (Hordeum vulgare L. cv. Morex). Four purification steps were used, carboxymethyl cellulose cation-exchange chromatography, chromatofocusing, size-exclusion chromatography, and electroelution from a polyacrylamide gel. The endopeptidase was most active at pH 4.5. It's isoelectric point (pI) was 4.4, as determined by isoelectric focusing, and it's SDS-PAGE molecular size was 31 kDa. The enzyme specifically hydrolyzed peptide bonds when the S2 site contained relatively large hydrophobic amino acids. The N-terminal amino acid sequence residues (1-9) of the 31-kDa endopeptidase had high homology to those of the EP-A and EP-B cysteine proteinases reported previously. The 31-kDa endopeptidase had a hydrolytic specificity similar to that of the Morex green malt 30-kDa endopeptidase we characterized previously, and also reacted with the antibody raised against the purified 30-kDa proteinase, but the two had different mobilities on non-denaturing PAGE. The hydrolytic specificities of both 30- and 31-kDa endopeptidases are such that both would very quickly cleave hordein (barley storage) proteins to small glutamine- and proline-rich peptides that could be quickly degraded to amino acids by barley exopeptidases.

  7. The 32 kDa Enamelin Undergoes Conformational Transitions upon Calcium Binding

    PubMed Central

    Fan, Daming; Lakshminarayanan, Rajamani; Moradian-Oldak, Janet

    2008-01-01

    The 32 kDa hydrophilic and acidic enamelin, the most stable cleavage fragment of the enamel specific glycoprotein, is believed to play vital roles in controlling crystal nucleation or growth during enamel biomineralization. Circular dichroism and Fourier transform infrared spectra demonstrate that the secondary structure of the 32 kDa enamelin has a high content of α-helix (81.5%). Quantitative analysis on the circular dichroism data revealed that the 32 kDa enamelin undergoes conformational changes with a structural preference to β-sheet as a function of calcium ions. We suggest that the increase of β-sheet conformation upon presence of Ca2+ may allow preferable interaction of the 32 kDa enamelin with apatite crystal surfaces during enamel biomineralization. The calcium association constant of the 32 kDa enamelin calculated from the fitting curve of ellipticity at 222 nm is Ka = 1.55 (±0.13) × 103 M−1, indicating a relatively low affinity. Our current biophysical studies on the 32 kDa enamelin structure provide novel insights towards understanding the enamelin-mineral interaction and subsequently the functions of enamelin during enamel formation. PMID:18508280

  8. OpenDA-WFLOW framework for improving hydrologic predictions using distributed hydrologic models

    NASA Astrophysics Data System (ADS)

    Weerts, Albrecht; Schellekens, Jaap; Kockx, Arno; Hummel, Stef

    2017-04-01

    Data assimilation (DA) holds considerable potential for improving hydrologic predictions (Liu et al., 2012) and increase the potential for early warning and/or smart water management. However, advances in hydrologic DA research have not yet been adequately or timely implemented in operational forecast systems to improve the skill of forecasts for better informed real-world decision making. The objective of this work is to highlight the development of a generic linkage of the open source OpenDA package and the open source community hydrologic modeling framework Openstreams/WFLOW and its application in operational hydrological forecasting on various spatial scales. The coupling between OpenDA and Openstreams/wflow framework is based on the emerging standard Basic Model Interface (BMI) as advocated by CSDMS using cross-platform webservices (i.e. Apache Thrift) developed by Hut et al. (2016). The potential application of the OpenDA-WFLOW for operational hydrologic forecasting including its integration with Delft-FEWS (used by more than 40 operational forecast centers around the world (Werner et al., 2013)) is demonstrated by the presented case studies. We will also highlight the possibility to give real-time insight into the working of the DA methods applied for supporting the forecaster as mentioned as one of the burning issues by Liu et al., (2012).

  9. In vitro degradation of the 32kDa PS II reaction centre protein

    SciTech Connect

    Eckenswiller, L.C.; Greenberg, B.M. )

    1989-04-01

    The 32kDa thylakoid membrane protein is an integral component of the PS II reaction centre. The protein, although stable in the dark, undergoes light dependent turnover. Light from the UV, visible and far-red spectral regions induce 32kDa protein degradation. To better understand 32kDa protein metabolism, an in vitro degradation system is being developed. It consists of isolated thylakoid membranes than contain radiolabelled protein. The 32kDa protein is actively and specifically degraded when the thylakoid preparation is exposed to UV or visible radiation. The protein is stable in the dark. The herbicides (atrazine and DCMU) inhibit degradation in the in vitro system as they do in vivo. Additionally, several methods of isolating thylakoids are being compared to optimize the 32kDa protein degradation reaction. The preparations will be evaluated based on their ability to permit light dependent degradation of the 32kDa protein without affecting the other membrane components.

  10. DaPKC-dependent phosphorylation of Crumbs is required for epithelial cell polarity in Drosophila

    PubMed Central

    Sotillos, Sol; Díaz-Meco, María Teresa; Caminero, Eva; Moscat, Jorge; Campuzano, Sonsoles

    2004-01-01

    Both in Drosophila and vertebrate epithelial cells, the establishment of apicobasal polarity requires the apically localized, membrane-associated Par-3–Par-6–aPKC protein complex. In Drosophila, this complex colocalizes with the Crumbs–Stardust (Sdt)–Pals1-associated TJ protein (Patj) complex. Genetic and molecular analyses suggest a functional relationship between them. We show, by overexpression of a kinase-dead Drosophila atypical PKC (DaPKC), the requirement for the kinase activity of DaPKC to maintain the position of apical determinants and to restrict the localization of basolateral ones. We demonstrate a novel physical interaction between the apical complexes, via direct binding of DaPKC to both Crb and Patj, and identify Crumbs as a phosphorylation target of DaPKC. This phosphorylation of Crumbs is functionally significant. Thus, a nonphosphorylatable Crumbs protein behaves in vivo as a dominant negative. Moreover, the phenotypic effect of overexpressing wild-type Crumbs is suppressed by reducing DaPKC activity. These results provide a mechanistic framework for the functional interaction between the Par-3–Par-6–aPKC and Crumbs–Sdt–Patj complexes based in the posttranslational modification of Crb by DaPKC. PMID:15302858

  11. SAGES TAVAC safety and effectiveness analysis: da Vinci ® Surgical System (Intuitive Surgical, Sunnyvale, CA).

    PubMed

    Tsuda, Shawn; Oleynikov, Dmitry; Gould, Jon; Azagury, Dan; Sandler, Bryan; Hutter, Matthew; Ross, Sharona; Haas, Eric; Brody, Fred; Satava, Richard

    2015-10-01

    The da Vinci(®) Surgical System (Intuitive Surgical, Sunnyvale, CA, USA) is a computer-assisted (robotic) surgical system designed to enable and enhance minimally invasive surgery. The Food and Drug Administration (FDA) has cleared computer-assisted surgical systems for use by trained physicians in an operating room environment for laparoscopic surgical procedures in general, cardiac, colorectal, gynecologic, head and neck, thoracic and urologic surgical procedures. There are substantial numbers of peer-reviewed papers regarding the da Vinci(®) Surgical System, and a thoughtful assessment of evidence framed by clinical opinion is warranted. The SAGES da Vinci(®) TAVAC sub-committee performed a literature review of the da Vinci(®) Surgical System regarding gastrointestinal surgery. Conclusions by the sub-committee were vetted by the SAGES TAVAC Committee and SAGES Executive Board. Following revisions, the document was evaluated by the TAVAC Committee and Executive Board again for final approval. Several conclusions were drawn based on expert opinion organized by safety, efficacy, and cost for robotic foregut, bariatric, hepatobiliary/pancreatic, colorectal surgery, and single-incision cholecystectomy. Gastrointestinal surgery with the da Vinci(®) Surgical System is safe and comparable, but not superior to standard laparoscopic approaches. Although clinically acceptable, its use may be costly for select gastrointestinal procedures. Current data are limited to the da Vinci(®) Surgical System; further analyses are needed.

  12. Trichosporon asahii secretes a 30-kDa aspartic peptidase.

    PubMed

    Valle, Roberta S; Ramos, Lívia S; Reis, Vanessa J; Ziccardi, Mariangela; Dornelas-Ribeiro, Marcos; Sodré, Cátia L; Branquinha, Marta H; Santos, André L S

    2017-12-01

    Trichosporon asahii is a fungal opportunistic pathogen that causes superficial and deep-seated infections presenting high mortality. Very little is known about the virulence attributes produced by this fungus. Herein, aspartic peptidase production was identified in Brazilian clinical isolates of T. asahii by different methodologies. Initially, T. asahii strain 250 (from skin lesion) was inoculated in both liquid and solid culture media containing bovine serum albumin (BSA) as the sole nitrogenous source. A translucent halo around the fungal colony was observed from the 5th day of culture. The cell-free culture supernatant revealed that soluble BSA was hydrolyzed along the growth, generating low molecular mass polypeptides as observed by electrophoresis. Subsequently, the secretions from four clinical strains of T. asahii were analyzed by BSA-SDS-PAGE and a single proteolytic band of 30-kDa was detected under acidic pH at 37°C. The secreted aspartic peptidase of T. asahii efficiently cleaved the cathepsin D peptide substrate, but not the substrates with specificity to HIV-1 peptidase and rennin. The capability to cleave either cathepsin D substrate in a fluorogenic assay or BSA immobilized within a gel matrix varied according to the T. asahii isolate. T. asahii extracellular peptidase activity was strongly inhibited by pepstatin A and HIV peptidase inhibitors, classifying it as an aspartic-type peptidase. Human serum albumin, mucin, non-immune immunoglobulin G and gelatin induced, in different levels, the secretion of this aspartic peptidase. With these results, T. asahii must be included in the list of many human fungal opportunistic pathogens able to secrete an aspartic-type peptidase. Copyright © 2017 Elsevier GmbH. All rights reserved.

  13. Leonardo da Vinci and the first hemodynamic observations.

    PubMed

    Martins e Silva, J

    2008-02-01

    Leonardo da Vinci was a genius whose accomplishments and ideas come down to us today, five centuries later, with the freshness of innovation and the fascination of discovery. This brief review begins with a summary of Leonardo's life and a description of the most important works of art that he bequeathed us, and then concentrates on his last great challenge. There was a point at which Leonardo's passion for art gave way to the study of human anatomy, not only to improve his drawing but to go beyond what had been simply a representation of form to understand the underlying functioning. Among his many interests, we focus on his study of the heart and blood vessels, which he observed carefully in animals and human autopsies, and reproduced in drawings of great quality with annotations of astonishing acuteness. The experience that he had acquired from observing the flow of water in currents and around obstacles, and the conclusions that he drew concerning hydrodynamics, were central to his interpretation of the mechanisms of the heart and of blood flow, to which he devoted much of his time between 1508 and 1513. From these studies, immortalized in drawings of great clarity, come what are acknowledged to be the first hemodynamic records, in which Leonardo demonstrates the characteristics of blood flow in the aorta and great vessels and the importance of blood reflux and the formation of eddies in the sinus in aortic valve his assiduous and careful observations, and his subsequent deductions, Leonardo put forward detailed findings on hemodynamic questions that advanced technology has only recently enabled us to confirm.

  14. Case study of Berengario da Carpi and Lorenzo de' Medici.

    PubMed

    Lippi, D

    2017-09-01

    Jacopo Berengario da Carpi (c.1460-c.1530) made several important advances in anatomy, being universally considered the founder of 'animated anatomy' (anatomia animata). In addition to being a famous anatomist, Berengario was also a highly regarded surgeon. One of his famous clients was Lorenzo de' Medici, Duke of Urbino (1492-1519). In 1517, Lorenzo suffered a skull injury from an harquebus shot and Berengario was asked to come to his bedside. Lorenzo's case gave Berengario the opportunity to write his Tractatus de Fractura Calve sive Cranei, published in Bologna by Gerolamo Benedetti in 1518. Berengario addressed his treatise to Lorenzo himself. This illustrated monograph was the most original neurosurgical treatise at that time, as Berengario was able to cite both from contemporary information and from his own direct observation all possible kinds of skull fracture, including the relationship between the site of the lesions and the resulting neurological effects. At the end of the book, Berengario explained and illustrated the surgical equipment to be used in each case, depicting a drill previously unseen in a medical volume and providing the recipe for a human dressing, a kind of poultice made of mummified human remins, to be applied regularly to wounds. Lorenzo de' Medici died in 1519 and was buried in the Church of San Lorenzo in Florence. His corpse was exhumed in 1875 and 1947. The casts of his skull made on those occasions are now preserved in the museums of Florence University, and clearly show evidence of the wound. Read more about the stories behind this masterpiece in an essay online. © 2017 BJS Society Ltd Published by John Wiley & Sons Ltd.

  15. Sequence and expression variations suggest an adaptive role for the DA1-like gene family in the evolution of soybeans.

    PubMed

    Zhao, Man; Gu, Yongzhe; He, Lingli; Chen, Qingshan; He, Chaoying

    2015-05-15

    The DA1 gene family is plant-specific and Arabidopsis DA1 regulates seed and organ size, but the functions in soybeans are unknown. The cultivated soybean (Glycine max) is believed to be domesticated from the annual wild soybeans (Glycine soja). To evaluate whether DA1-like genes were involved in the evolution of soybeans, we compared variation at both sequence and expression levels of DA1-like genes from G. max (GmaDA1) and G. soja (GsoDA1). Sequence identities were extremely high between the orthologous pairs between soybeans, while the paralogous copies in a soybean species showed a relatively high divergence. Moreover, the expression variation of DA1-like paralogous genes in soybean was much greater than the orthologous gene pairs between the wild and cultivated soybeans during development and challenging abiotic stresses such as salinity. We further found that overexpressing GsoDA1 genes did not affect seed size. Nevertheless, overexpressing them reduced transgenic Arabidopsis seed germination sensitivity to salt stress. Moreover, most of these genes could improve salt tolerance of the transgenic Arabidopsis plants, corroborated by a detection of expression variation of several key genes in the salt-tolerance pathways. Our work suggested that expression diversification of DA1-like genes is functionally associated with adaptive radiation of soybeans, reinforcing that the plant-specific DA1 gene family might have contributed to the successful adaption to complex environments and radiation of the plants.

  16. Myelin management by the 18.5–kDa and 21.5–kDa classic myelin basic protein isoforms

    PubMed Central

    Harauz, George; Boggs, Joan M.

    2013-01-01

    The classic myelin basic protein (MBP) splice isoforms range in nominal molecular mass from 14 to 21.5 kDa, and arise from the gene in the oligodendrocyte lineage (Golli) in maturing oligodendrocytes. The 18.5-kDa isoform that predominates in adult myelin adheres the cytosolic surfaces of oligodendrocyte membranes together, and forms a two-dimensional molecular sieve restricting protein diffusion into compact myelin. However, this protein has additional roles including cytoskeletal assembly and membrane extension, binding to SH3-domains, participation in Fyn-mediated signaling pathways, sequestration of phosphoinositides, and maintenance of calcium homeostasis. Of the diverse post-translational modifications of this isoform, phosphorylation is the most dynamic, and modulates 18.5-kDa MBP’s protein-membrane and protein-protein interactions, indicative of a rich repertoire of functions. In developing and mature myelin, phosphorylation can result in microdomain or even nuclear targeting of the protein, supporting the conclusion that 18.5-kDa MBP has significant roles beyond membrane adhesion. The full-length, early-developmental 21.5-kDa splice isoform is predominantly karyophilic due to a non-traditional P-Y nuclear localization signal, with effects such as promotion of oligodendrocyte proliferation. We discuss in vitro and recent in vivo evidence for multifunctionality of these classic basic proteins of myelin, and argue for a systematic evaluation of the temporal and spatial distributions of these protein isoforms, and their modified variants, during oligodendrocyte differentiation. PMID:23398367

  17. Multiquadrant robotic colorectal surgery: the da Vinci Xi vs Si comparison.

    PubMed

    Protyniak, Bogdan; Jorden, Jeffrey; Farmer, Russell

    2017-03-08

    The newly introduced da Vinci Xi Surgical System hopes to address the shortcomings of its predecessor, specifically robotic arm restrictions and difficulty working in multiple quadrants. We compare the two robot platforms in multiquadrant surgery at a major colorectal referral center. Forty-four patients in the da Vinci Si group and 26 patients in the Xi group underwent sigmoidectomy or low anterior resection between 2014 and 2016. Patient demographics, operative variables, and postoperative outcomes were compared using descriptive statistics. Both groups were similar in age, sex, BMI, pelvic surgeries, and ASA class. Splenic flexure was mobilized in more (p = 0.045) da Vinci Xi cases compared to da Vinci Si both for sigmoidectomy (50 vs 15.4%) and low anterior resection (60 vs 29%). There was no significant difference in operative time (219.9 vs 224.7 min; p = 0.640), blood loss (170.0 vs 188.1 mL; p = 0.289), length of stay (5.7 vs 6 days; p = 0.851), or overall complications (26.9 vs 22.7%; p = 0.692) between the da Vinci Xi and Si groups, respectively. Single-dock multiquadrant robotic surgery, measured by splenic flexure mobilization with concomitant pelvic dissection, was more frequently performed using the da Vinci Xi platform with no increase in operative time, bleeding, or postoperative complications. The new platform provides surgeons an easier alternative to the da Vinci Si dual docking or combined robotic/laparoscopic multiquadrant surgery.

  18. Maize 27 kDa gamma-zein is a potential allergen for early weaned pigs.

    PubMed

    Krishnan, Hari B; Kerley, Monty S; Allee, Gary L; Jang, Sungchan; Kim, Won-Seok; Fu, Chunjiang J

    2010-06-23

    Soybean and maize are extensively used in animal feed, primarily in poultry, swine, and cattle diets. Soybean meal can affect pig performance in the first few weeks following weaning and elicit specific antibodies in weaned piglets. Though maize is a major component of pig feed, it is not known if any of the maize proteins can elicit immunological response in young pigs. In this study, we have identified a prominent 27 kDa protein from maize as an immunodominant protein in young pigs. This protein, like some known allergens, exhibited resistance to pepsin digestion in vitro. Several lines of evidence identify the immunodominant 27 kDa protein as a gamma-zein, a maize seed storage protein. First, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of different solubility classes of maize seed proteins revealed the presence of an abundant 27 kDa protein in the prolamin (zein) fraction. Antibodies raised against the purified maize 27 kDa gamma-zein also reacted against the same protein recognized by the young pig serum. Additionally, matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the peptides generated by trypsin digestion of the immunodominant 27 kDa protein showed significant homology to the maize 27 kDa gamma-zein. Since eliminating the allergenic protein will have a great impact on the nutritive value of the maize meal and expand its use in the livestock industry, it will be highly desirable to develop maize cultivars completely lacking the 27 kDa allergenic protein.

  19. Crystal structure of Bombyx mori lipoprotein 6: comparative structural analysis of the 30-kDa lipoprotein family.

    PubMed

    Pietrzyk, Agnieszka J; Bujacz, Anna; Łochynska, Malgorzata; Jaskolski, Mariusz; Bujacz, Grzegorz

    2014-01-01

    The 30-kDa lipoprotein (LP) family of mulberry silkworm comprises major hemolymph proteins specific to the fifth instar larvae. The family consists of 46 members, 24 of which are referred to as typical 30-kDa LPs. To date, two crystal structures of 30-kDa LPs from Bombyx mori have been described (Bmlp3 and Bmlp7). Here, we present the crystal structure of Bmlp6, another 30-kDa LP member. Bmlp6 is comprised of two domains characteristic of this family, the VHS-type N-terminal domain and β-trefoil C-terminal domain. The structures of the three 30-kDa LPs have been compared and a number of differences are noted, including loop conformation, the surface electrostatic potential, and the potential binding cavities. We discuss the observed structural differences in the light of the potential different roles of the particular 30-kDa LP members in silkworm physiology.

  20. Nucleus-localized 21.5-kDa myelin basic protein promotes oligodendrocyte proliferation and enhances neurite outgrowth in coculture, unlike the plasma membrane-associated 18.5-kDa isoform.

    PubMed

    Smith, Graham S T; Samborska, Bożena; Hawley, Steven P; Klaiman, Jordan M; Gillis, Todd E; Jones, Nina; Boggs, Joan M; Harauz, George

    2013-03-01

    The classic myelin basic protein (MBP) family of central nervous system (CNS) myelin arises from transcription start site 3 of the Golli (gene of oligodendrocyte lineage) complex and comprises splice isoforms ranging in nominal molecular mass from 14 kDa to (full-length) 21.5 kDa. We have determined here a number of distinct functional differences between the major 18.5-kDa and minor 21.5-kDa isoforms of classic MBP with respect to oligodendrocyte (OLG) proliferation. We have found that, in contrast to 18.5-kDa MBP, 21.5-kDa MBP increases proliferation of early developmental immortalized N19-OLGs by elevating the levels of phosphorylated ERK1/2 and Akt1 kinases and of ribosomal protein S6. Coculture of N2a neuronal cells with N19-OLGs transfected with the 21.5-kDa isoform (or conditioned medium from), but not the 18.5-kDa isoform, caused the N2a cells to have increased neurite outgrowth and process branching complexity. These roles were dependent on subcellular localization of 21.5-kDa MBP to the nucleus and on the exon II-encoded segment, suggesting that the nuclear localization of early minor isoforms of MBP may play a crucial role in regulating and/or initiating myelin and neuronal development in the mammalian CNS.

  1. Therapeutic Potential of Dioscorea Extract (DA-9801) in Comparison with Alpha Lipoic Acid on the Peripheral Nerves in Experimental Diabetes.

    PubMed

    Jin, Heung Yong; Kim, Sun Hee; Yu, Hea Min; Baek, Hong Sun; Park, Tae Sun

    2013-01-01

    DA-9801, a mixture of extracts from Dioscorea japonica Thunb. and Dioscorea nipponica Makino, was reported to have neurotrophic activity. Therefore, we investigated the therapeutic potential of DA-9801, in comparison with alpha lipoic acid (ALA), for peripheral nerves preservation in experimental diabetes. Experimental animals were divided into 4 groups, and each group was designated according to the type of treatment administered as follows: normal, DM, DM+DA-9801, and DM+ALA. After 16 weeks, response thresholds to tactile and thermal stimuli were higher in DM+DA-9801 group than in nontreated DM group. This degree of increase in DM+DA-9801 group indicates more therapeutic potency of DA-9801 than ALA. Western blot analysis showed more significant increase in NGF and decrease in TNF-α and IL-6 in DM+DA-9801 group than in DM or DM+ALA groups (P < 0.05). IENF density was reduced less significantly in the DM+DA-9801 group than in other DM groups (7.61 ± 0.32, 4.2 ± 0.26, and 6.5 ± 0.30 in DM+DA-9801, DM, and DM+ALA, resp., P < 0.05). Mean myelinated axonal area in the sciatic nerves was significantly greater in DM+DA-9801 group than in other DM groups (69.2 ± 5.76, 54.0 ± 6.32, and 63.1 ± 5.41 in DM+DA-9801, DM, and DM+ALA, resp., P < 0.05). Results of this study demonstrated potential therapeutic applications of DA-9801 for the treatment of diabetic peripheral neuropathy.

  2. 82-kDa choline acetyltransferase and SATB1 localize to β-amyloid induced matrix attachment regions

    PubMed Central

    Winick-Ng, Warren; Caetano, Fabiana A.; Winick-Ng, Jennifer; Morey, Trevor M.; Heit, Bryan; Rylett, R. Jane

    2016-01-01

    The M-transcript of human choline acetyltransferase (ChAT) produces an 82-kDa protein (82-kDa ChAT) that concentrates in nuclei of cholinergic neurons. We assessed the effects of acute exposure to oligomeric amyloid-β1–42 (Aβ1–42) on 82-kDa ChAT disposition in SH-SY5Y neural cells, finding that acute exposure to Aβ1–42 results in increased association of 82-kDa ChAT with chromatin and formation of 82-kDa ChAT aggregates in nuclei. When measured by chromatin immunoprecipitation with next-generation sequencing (ChIP-seq), we identified that Aβ1–42 -exposure increases 82-kDa ChAT association with gene promoters and introns. The Aβ1–42 -induced 82-kDa ChAT aggregates co-localize with special AT-rich binding protein 1 (SATB1), which anchors DNA to scaffolding/matrix attachment regions (S/MARs). SATB1 had a similar genomic association as 82-kDa ChAT, with both proteins associating with synapse and cell stress genes. After Aβ1–42 -exposure, both SATB1 and 82-kDa ChAT are enriched at the same S/MAR on the APP gene, with 82-kDa ChAT expression attenuating an increase in an isoform-specific APP mRNA transcript. Finally, 82-kDa ChAT and SATB1 have patterned genomic association at regions enriched with S/MAR binding motifs. These results demonstrate that 82-kDa ChAT and SATB1 play critical roles in the response of neural cells to acute Aβ -exposure. PMID:27052102

  3. An Introduction to the DA-T Gibbs Sampler for the Two-Parameter Logistic (2PL) Model and beyond

    ERIC Educational Resources Information Center

    Maris, Gunter; Bechger, Timo M.

    2005-01-01

    The DA-T Gibbs sampler is proposed by Maris and Maris (2002) as a Bayesian estimation method for a wide variety of "Item Response Theory (IRT) models". The present paper provides an expository account of the DA-T Gibbs sampler for the 2PL model. However, the scope is not limited to the 2PL model. It is demonstrated how the DA-T Gibbs…

  4. A Influência da Escola e da Mídia na Obtenção dos Conhecimentos Astronômicos

    NASA Astrophysics Data System (ADS)

    da Cunha, W. S.; Voelzke, M. R.; Amaral, L. H.

    2006-08-01

    O universo em que vivemos possui segredos jamais imaginados pelo homem, fruto da pequena proporção que ocupamos diante de um cosmos infinito. A astronomia funciona como um instrumento que sacia a sede humana em desvendar esses segredos, estudando as galáxias, quasares, pulsares além da existência de todos os elementos que preenchem o espaço cósmico, o vácuo, não mais vazio como se pensou em outros tempos. A mídia explora esse campo de forma mística e também científica. A difusão adequada e dentro dos padrões educacionais vigentes, de conceitos astronômicos, se faz necessária, devido sua presença nos conteúdos escolares, centros de ciência e na mídia, de modo geral. A reforma do Currículo Básico da Escola Pública da maioria dos Estados brasileiros introduziu conceitos astronômicos desde a pré-escola até o Ensino Médio. Num estudo realizado com 1.032 alunos do Ensino Médio de seis escolas estaduais, revelou-se através da aplicação de um questionário que abordava conheciment os básicos em astronomia, como estes foram adquiridos e qual era a infra-estrutura oferecida pela escola: para 66,2% dos entrevistados o professor já fez alguma apresentação a respeito de astronomia. O presente estudo revelou também que 44,5% dos alunos entrevistados adquiriram seus conhecimentos astronômicos na escola, 28,1% através da televisão e 27,4% afirmaram que seus conhecimentos foram obtidos através de outros tipos de mídia (internet, filmes e revistas). Embora o estudo revele claramente que há certo equilíbrio na forma em que os alunos obtiveram seus conhecimentos astronômicos, a mídia ainda explora e muitas vezes desperta o interesse pela astronomia com assuntos que não fazem parte de nosso cotidiano como, por exemplo, o homem na Lua, os projetos espaciais e a primeira viagem de um astronauta brasileiro no espaço. Porém, em certos casos, a mídia pode promover conhecimentos incompletos ou mesmo equivocados sobre os conceitos que envolvam

  5. Morphogenetic Studies of the Drosophila DA1 Ventral Olfactory Projection Neuron.

    PubMed

    Shen, Hung-Chang; Wei, Jia-Yi; Chu, Sao-Yu; Chung, Pei-Chi; Hsu, Tsai-Chi; Yu, Hung-Hsiang

    2016-01-01

    In the Drosophila olfactory system, odorant information is sensed by olfactory sensory neurons and relayed from the primary olfactory center, the antennal lobe (AL), to higher olfactory centers via olfactory projection neurons (PNs). A major portion of the AL is constituted with dendrites of four groups of PNs, anterodorsal PNs (adPNs), lateral PNs (lPNs), lateroventral PNs (lvPNs) and ventral PNs (vPNs). Previous studies have been focused on the development and function of adPNs and lPNs, while the investigation on those of lvPNs and vPNs received less attention. Here, we study the molecular and cellular mechanisms underlying the morphogenesis of a putative male-pheromone responding vPN, the DA1 vPN. Using an intersection strategy to remove background neurons labeled within a DA1 vPN-containing GAL4 line, we depicted morphological changes of the DA1 vPN that occurs at the pupal stage. We then conducted a pilot screen using RNA interference knock-down approach to identify cell surface molecules, including Down syndrome cell adhesion molecule 1 and Semaphorin-1a, that might play essential roles for the DA1 vPN morphogenesis. Taken together, by revealing molecular and cellular basis of the DA1 vPN morphogenesis, we should provide insights into future comprehension of how vPNs are assembled into the olfactory neural circuitry.

  6. Morphogenetic Studies of the Drosophila DA1 Ventral Olfactory Projection Neuron

    PubMed Central

    Yu, Hung-Hsiang

    2016-01-01

    In the Drosophila olfactory system, odorant information is sensed by olfactory sensory neurons and relayed from the primary olfactory center, the antennal lobe (AL), to higher olfactory centers via olfactory projection neurons (PNs). A major portion of the AL is constituted with dendrites of four groups of PNs, anterodorsal PNs (adPNs), lateral PNs (lPNs), lateroventral PNs (lvPNs) and ventral PNs (vPNs). Previous studies have been focused on the development and function of adPNs and lPNs, while the investigation on those of lvPNs and vPNs received less attention. Here, we study the molecular and cellular mechanisms underlying the morphogenesis of a putative male-pheromone responding vPN, the DA1 vPN. Using an intersection strategy to remove background neurons labeled within a DA1 vPN-containing GAL4 line, we depicted morphological changes of the DA1 vPN that occurs at the pupal stage. We then conducted a pilot screen using RNA interference knock-down approach to identify cell surface molecules, including Down syndrome cell adhesion molecule 1 and Semaphorin-1a, that might play essential roles for the DA1 vPN morphogenesis. Taken together, by revealing molecular and cellular basis of the DA1 vPN morphogenesis, we should provide insights into future comprehension of how vPNs are assembled into the olfactory neural circuitry. PMID:27163287

  7. Low strain, long life creep fatigue of AF2-1DA and INCO 718

    NASA Technical Reports Server (NTRS)

    Thakker, A. B.; Cowles, B. A.

    1983-01-01

    Two aircraft turbine disk alloys, GATORIZED AF2-DA and INCO 718 were evaluated for their low strain long life creep-fatigue behavior. Static (tensile and creep rupture) and cyclic properties of both alloys were characterized. The cntrolled strain LCF tests were conducted at 760 C (1400 F) and 649 C (1200 F) for AF2-1DA and INCO 718, respectively. Hold times were varied for tensile, compressive and tensile/compressive strain dwell (relaxation) tests. Stress (creep) hold behavior of AF2-1DA was also evaluated. Generally, INCO 718 exhibited more pronounced reduction in cyclic life due to hold than AF2-1DA. The percent reduction in life for both alloys for strain dwell tests was greater at low strain ranges (longer life regime). Changing hold time from 0 to 0.5, 2.0 and 15.0 min. resulted in corresponding reductions in life. The continuous cycle and cyclic/dwell initiation failure mechanism was predominantly transgranular for AF2-1DA and intergranular for INCO 718.

  8. Color constrains depth in da Vinci stereopsis for camouflage but not occlusion.

    PubMed

    Wardle, Susan G; Gillam, Barbara J

    2013-12-01

    Monocular regions that occur with binocular viewing of natural scenes can produce a strong perception of depth--"da Vinci stereopsis." They occur either when part of the background is occluded in one eye, or when a nearer object is camouflaged against a background surface in one eye's view. There has been some controversy over whether da Vinci depth is constrained by geometric or ecological factors. Here we show that the color of the monocular region constrains the depth perceived from camouflage, but not occlusion, as predicted by ecological considerations. Quantitative depth was found in both cases, but for camouflage only when the color of the monocular region matched the binocular background. Unlike previous reports, depth failed even when nonmatching colors satisfied conditions for perceptual transparency. We show that placing a colored line at the boundary between the binocular and monocular regions is sufficient to eliminate depth from camouflage. When both the background and the monocular region contained vertical contours that could be fused, some observers appeared to use fusion, and others da Vinci constraints, supporting the existence of a separate da Vinci mechanism. The results show that da Vinci stereopsis incorporates color constraints and is more complex than previously assumed.

  9. Dopamine DA1 receptors on vascular smooth muscle cells are regulated by glucocorticoid and sodium chloride.

    PubMed

    Yasunari, K; Kohno, M; Yokokawa, K; Horio, T; Takeda, T

    1994-09-01

    The modulation of dopamine DA1 receptors of cultured rat renal arterial smooth muscle cells by glucocorticoid and sodium chloride was studied. At a concentration of 10 nM, the synthetic glucocorticoid dexamethasone increased maximum receptor binding but had no effect on the dissociation constant. However, the maximum binding of [3H]Sch-23390 in cells treated with 100 mM sodium chloride did not change. However, the dissociation constant for DA1 receptor was increased by adding sodium chloride. The glucocorticoid effect on DA1 of arterial smooth muscle cells became apparent after hours of incubation in the presence of the steroid and was significantly inhibited by cycloheximide (10 micrograms/ml) or by the glucocorticoid receptor antagonist RU-38486, indicating that the effect required protein synthesis through glucocorticoid receptors. Treatment of cells with 1 microM dexamethasone for 24 h increased basal and DA1-stimulated adenylate cyclase activity. Basal adenylate cyclase was decreased by sodium chloride in a dose-dependent manner. These results suggest differential control of DA1 receptors on vascular smooth muscle cells by glucocorticoid or sodium chloride.

  10. [Discrimination of wood biological decay by NIR and partial least squares discriminant analysis (PLS-DA)].

    PubMed

    Yang, Zhong; Ren, Hai-Qing; Jiang, Ze-Hui

    2008-04-01

    Extensive research has demonstrated that near infrared spectroscopy (NIR) and partial least squares discriminant analysis (PLS-DA) can be used to rapidly discriminate or detect a wide variety of food, medicine and agricultural products. The use of NIR coupled with PLS-DA to detect wood biological decay was investigated in the present paper. The results showed that the correlation between the predicted category variable of calibration and validation and the measured category variable is significant with a correlation coefficient (r) over 0.94 and low SEC and SEP (< 0.17); the discriminant accuracy for the non-decay, white-rot and brown-rot decay samples are 100% (deviation < 0.5) by the PLS-DA model based on the test set of samples; the discriminant accuracy by PLS-DA model is better than that by SIMCA model. It's suggested that NIR spectroscopy coupled with PLS-DA could be used to rapidly detect wood biological decay.

  11. In vitro study on the interaction between the 32 kDa enamelin and amelogenin

    PubMed Central

    Fan, Daming; Du, Chang; Sun, Zhi; Lakshminarayanan, Rajamani; Moradian-Oldak, Janet

    2015-01-01

    Enamel extracelluar matrix components play vital roles in controlling crystal nucleation and growth during enamel formation. We investigated the interaction between the 32 kDa enamelin fragment and amelogenin using immunochemical and biophysical methods. Immunoprecipitation studies revealed that the 32 kDa enamelin and amelogenin eluted together from a Protein A column. Dynamic light scattering results showed that the 32 kDa enamelin had a profound effect on amelogenin assembly at pH 8.0, causing partial dissociation of the nanospheres, in a dose-dependent manner. The appearance of an isodichroic point and the shifting and intensity decrease of the ellipticity minima in the circular dichroism spectra of amelogenin following the addition of the 32 kDa enamelin were indicative of conformational changes in amelogenin and of a direct interaction between the two macromolecules. Our results collectively demonstrate that the 32 kDa enamelin has a direct interaction with amelogenin in vitro. Our current studies provide novel insights into understanding possible cooperation between enamelin and amelogenin in macromolecular self-assembly and in controlling enamel mineral formation. PMID:19263522

  12. PanDA: Exascale Federation of Resources for the ATLAS Experiment at the LHC

    NASA Astrophysics Data System (ADS)

    Barreiro Megino, Fernando; Caballero Bejar, Jose; De, Kaushik; Hover, John; Klimentov, Alexei; Maeno, Tadashi; Nilsson, Paul; Oleynik, Danila; Padolski, Siarhei; Panitkin, Sergey; Petrosyan, Artem; Wenaus, Torre

    2016-02-01

    After a scheduled maintenance and upgrade period, the world's largest and most powerful machine - the Large Hadron Collider(LHC) - is about to enter its second run at unprecedented energies. In order to exploit the scientific potential of the machine, the experiments at the LHC face computational challenges with enormous data volumes that need to be analysed by thousand of physics users and compared to simulated data. Given diverse funding constraints, the computational resources for the LHC have been deployed in a worldwide mesh of data centres, connected to each other through Grid technologies. The PanDA (Production and Distributed Analysis) system was developed in 2005 for the ATLAS experiment on top of this heterogeneous infrastructure to seamlessly integrate the computational resources and give the users the feeling of a unique system. Since its origins, PanDA has evolved together with upcoming computing paradigms in and outside HEP, such as changes in the networking model, Cloud Computing and HPC. It is currently running steadily up to 200 thousand simultaneous cores (limited by the available resources for ATLAS), up to two million aggregated jobs per day and processes over an exabyte of data per year. The success of PanDA in ATLAS is triggering the widespread adoption and testing by other experiments. In this contribution we will give an overview of the PanDA components and focus on the new features and upcoming challenges that are relevant to the next decade of distributed computing workload management using PanDA.

  13. Interferon gamma induces the myristoylation of a 48-kDa protein in macrophages.

    PubMed Central

    Aderem, A A; Marratta, D E; Cohn, Z A

    1988-01-01

    The lymphokine interferon gamma (IFN-gamma) induces the selective myristoylation of a macrophage protein with an apparent molecular mass of 48 kDa. The myristic acid-protein bond is resistant to treatment with hydroxylamine, suggesting that the fatty acid moiety is in an amide linkage. As little as 1 unit of IFN-gamma per ml induces the myristoylation of the 48-kDa protein, with half-maximal myristoylation being observed with 4 units/ml. The effect is observed within 1 hr after exposure to IFN-gamma and is maximal by 3-4 hr, after which it declines. IFN-alpha does not induce the myristoylation of the 48-kDa protein, and IFN-beta does so very poorly. Neither IFN-alpha nor IFN-beta has any effect on IFN-gamma-induced myristoylation of the 48-kDa protein. The 48-kDa protein is constitutively myristoylated in murine macrophages that have been activated in vivo by intraperitoneal injection of Corynebacterium parvum, suggesting that it may be an early intermediate in the activation of macrophages. Images PMID:3137568

  14. Influence of 120 kDa Pyruvate:Ferredoxin Oxidoreductase on Pathogenicity of Trichomonas vaginalis.

    PubMed

    Song, Hyun-Ouk

    2016-02-01

    Trichomonas vaginalis is a flagellate protozoan parasite and commonly infected the lower genital tract in women and men. Iron is a known nutrient for growth of various pathogens, and also reported to be involved in establishment of trichomoniasis. However, the exact mechanism was not clarified. In this study, the author investigated whether the 120 kDa protein of T. vaginalis may be involved in pathogenicity of trichomonads. Antibodies against 120 kDa protein of T. vaginalis, which was identified as pyruvate:ferredoxin oxidoreductase (PFOR) by peptide analysis of MALDI-TOF-MS, were prepared in rabbits. Pretreatment of T. vaginalis with anti-120 kDa Ab decreased the proliferation and adherence to vaginal epithelial cells (MS74) of T. vaginalis. Subcutaneous tissue abscess in anti-120 kDa Ab-treated T. vaginalis-injected mice was smaller in size than that of untreated T. vaginalis-infected mice. Collectively, the 120 kDa protein expressed by iron may be involved in proliferation, adhesion to host cells, and abscess formation, thereby may influence on the pathogenicity of T. vaginalis.

  15. A constitutively expressed 36 kDa exochitinase from Bacillus thuringiensis HD-1.

    PubMed

    Arora, Naresh; Ahmad, Tarannum; Rajagopal, R; Bhatnagar, Raj K

    2003-08-01

    A 36 kDa chitinase was purified by ion exchange and gel filtration chromatography from the culture supernatant of Bacillus thuringiensis HD-1. The chitinase production was independent of the presence of chitin in the growth medium and was produced even in the presence of glucose. The purified chitinase was active at acidic pH, had an optimal activity at pH 6.5, and showed maximum activity at 65 degrees C. Of the various substrates, the enzyme catalyzed the hydrolysis of the disaccharide 4-MU(GlnAc)(2) most efficiently and was therefore classified as an exochitinase. The sequence of the tryptic peptides showed extensive homology with Bacillus cereus 36 kDa exochitinase. The 1083 bp open reading frame encoding 36 kDa chitinase was amplified with primers based on the gene sequence of B. cereus 36 kDa exochitinase. The deduced amino-acid sequence showed that the protein contained an N-terminal signal peptide and consisted of a single catalytic domain. The two conserved signature sequences characteristic of family 18 chitinases were mapped at positions 105-109 and 138-145 of Chi36. The recombinant chitinase was expressed in a catalytically active form in Escherichia coli in the vector pQE-32. The expressed 36 kDa chitinase potentiated the insecticidal effect of the vegetative insecticidal protein (Vip) when used against neonate larvae of Spodoptera litura.

  16. Preclinical and Clinical Studies Demonstrate That the Proprietary Herbal Extract DA-5512 Effectively Stimulates Hair Growth and Promotes Hair Health

    PubMed Central

    Yu, Jae Young; Gupta, Biki; Park, Hyoung Geun; Son, Miwon; Jun, Joon-Ho; Yong, Chul Soon

    2017-01-01

    The proprietary DA-5512 formulation comprises six herbal extracts from traditional oriental plants historically associated with therapeutic and other applications related to hair. Here, we investigated the effects of DA-5512 on the proliferation of human dermal papilla cells (hDPCs) in vitro and on hair growth in C57BL/6 mice and conducted a clinical study to evaluate the efficacy and safety of DA-5512. DA-5512 significantly enhanced the viability of hDPCs in a dose-dependent manner (p < 0.05), and 100 ppm of DA-5512 and 1 μM minoxidil (MXD) significantly increased the number of Ki-67-positive cells, compared with the control group (p < 0.05). MXD (3%) and DA-5512 (1%, 5%) significantly stimulated hair growth and increased the number and length of hair follicles (HFs) versus the controls (each p < 0.05). The groups treated with DA-5512 exhibited hair growth comparable to that induced by MXD. In clinical study, we detected a statistically significant increase in the efficacy of DA-5512 after 16 weeks compared with the groups treated with placebo or 3% MXD (p < 0.05). In conclusion, DA-5512 might promote hair growth and enhance hair health and can therefore be considered an effective option for treating hair loss. PMID:28539964

  17. Preclinical and Clinical Studies Demonstrate That the Proprietary Herbal Extract DA-5512 Effectively Stimulates Hair Growth and Promotes Hair Health.

    PubMed

    Yu, Jae Young; Gupta, Biki; Park, Hyoung Geun; Son, Miwon; Jun, Joon-Ho; Yong, Chul Soon; Kim, Jeong Ah; Kim, Jong Oh

    2017-01-01

    The proprietary DA-5512 formulation comprises six herbal extracts from traditional oriental plants historically associated with therapeutic and other applications related to hair. Here, we investigated the effects of DA-5512 on the proliferation of human dermal papilla cells (hDPCs) in vitro and on hair growth in C57BL/6 mice and conducted a clinical study to evaluate the efficacy and safety of DA-5512. DA-5512 significantly enhanced the viability of hDPCs in a dose-dependent manner (p < 0.05), and 100 ppm of DA-5512 and 1 μM minoxidil (MXD) significantly increased the number of Ki-67-positive cells, compared with the control group (p < 0.05). MXD (3%) and DA-5512 (1%, 5%) significantly stimulated hair growth and increased the number and length of hair follicles (HFs) versus the controls (each p < 0.05). The groups treated with DA-5512 exhibited hair growth comparable to that induced by MXD. In clinical study, we detected a statistically significant increase in the efficacy of DA-5512 after 16 weeks compared with the groups treated with placebo or 3% MXD (p < 0.05). In conclusion, DA-5512 might promote hair growth and enhance hair health and can therefore be considered an effective option for treating hair loss.

  18. The rare DAT coding variant Val559 perturbs DA neuron function, changes behavior, and alters in vivo responses to psychostimulants.

    PubMed

    Mergy, Marc A; Gowrishankar, Raajaram; Gresch, Paul J; Gantz, Stephanie C; Williams, John; Davis, Gwynne L; Wheeler, C Austin; Stanwood, Gregg D; Hahn, Maureen K; Blakely, Randy D

    2014-11-04

    Despite the critical role of the presynaptic dopamine (DA) transporter (DAT, SLC6A3) in DA clearance and psychostimulant responses, evidence that DAT dysfunction supports risk for mental illness is indirect. Recently, we identified a rare, nonsynonymous Slc6a3 variant that produces the DAT substitution Ala559Val in two male siblings who share a diagnosis of attention-deficit hyperactivity disorder (ADHD), with other studies identifying the variant in subjects with bipolar disorder (BPD) and autism spectrum disorder (ASD). Previously, using transfected cell studies, we observed that although DAT Val559 displays normal total and surface DAT protein levels, and normal DA recognition and uptake, the variant transporter exhibits anomalous DA efflux (ADE) and lacks capacity for amphetamine (AMPH)-stimulated DA release. To pursue the significance of these findings in vivo, we engineered DAT Val559 knock-in mice, and here we demonstrate in this model the presence of elevated extracellular DA levels, altered somatodendritic and presynaptic D2 DA receptor (D2R) function, a blunted ability of DA terminals to support depolarization and AMPH-evoked DA release, and disruptions in basal and psychostimulant-evoked locomotor behavior. Together, our studies demonstrate an in vivo functional impact of the DAT Val559 variant, providing support for the ability of DAT dysfunction to impact risk for mental illness.

  19. Fluorescence spectroscopy coupled with PARAFAC and PLS DA for characterization and classification of honey.

    PubMed

    Lenhardt, Lea; Bro, Rasmus; Zeković, Ivana; Dramićanin, Tatjana; Dramićanin, Miroslav D

    2015-05-15

    Fluorescence spectroscopy coupled with parallel factor analysis (PARAFAC) and Partial least squares Discriminant Analysis (PLS DA) were used for characterization and classification of honey. Excitation emission spectra were obtained for 95 honey samples of different botanical origin (acacia, sunflower, linden, meadow, and fake honey) by recording emission from 270 to 640 nm with excitation in the range of 240-500 nm. The number of fluorophores present in honey, excitation and emission spectra of each fluorophore, and their relative concentration are determined using a six-component PARAFAC model. Emissions from phenolic compounds and Maillard reaction products exhibited the largest difference among classes of honey of different botanical origin. The PLS DA classification model, constructed from PARAFAC model scores, detected fake honey samples with 100% sensitivity and specificity. Honey samples were also classified using PLS DA with errors of 0.5% for linden, 10% for acacia, and about 20% for both sunflower and meadow mix.

  20. gLExec Integration with the ATLAS PanDA Workload Management System

    NASA Astrophysics Data System (ADS)

    Karavakis, E.; Barreiro, F.; Campana, S.; De, K.; Di Girolamo, A.; Litmaath, M.; Maeno, T.; Medrano, R.; Nilsson, P.; Wenaus, T.

    2015-12-01

    ATLAS user jobs are executed on Worker Nodes (WNs) by pilots sent to sites by pilot factories. This paradigm serves to allow a high job reliability and although it has clear advantages, such as making the working environment homogeneous, the approach presents security and traceability challenges. To address these challenges, gLExec can be used to let the payloads for each user be executed under a different UNIX user id that uniquely identifies the ATLAS user. This paper describes the recent improvements and evolution of the security model within the ATLAS PanDA system, including improvements in the PanDA pilot, in the PanDA server and their integration with MyProxy, a credential caching system that entitles a person or a service to act in the name of the issuer of the credential. Finally, it presents results from ATLAS user jobs running with gLExec and describes the deployment campaign within ATLAS.

  1. The Da Vinci European BioBank: A Metabolomics-Driven Infrastructure

    PubMed Central

    Carotenuto, Dario; Luchinat, Claudio; Marcon, Giordana; Rosato, Antonio; Turano, Paola

    2015-01-01

    We present here the organization of the recently-constituted da Vinci European BioBank (daVEB, https://www.davincieuropeanbiobank.org/it). The biobank was created as an infrastructure to support the activities of the Fiorgen Foundation (http://www.fiorgen.net/), a nonprofit organization that promotes research in the field of pharmacogenomics and personalized medicine. The way operating procedures concerning samples and data have been developed at daVEB largely stems from the strong metabolomics connotation of Fiorgen and from the involvement of the scientific collaborators of the foundation in international/European projects aimed to tackle the standardization of pre-analytical procedures and the promotion of data standards in metabolomics. PMID:25913579

  2. Evolution of the ATLAS PanDA Production and Distributed Analysis System

    NASA Astrophysics Data System (ADS)

    Maeno, T.; De, K.; Wenaus, T.; Nilsson, P.; Walker, R.; Stradling, A.; Fine, V.; Potekhin, M.; Panitkin, S.; Compostella, G.

    2012-12-01

    The PanDA (Production and Distributed Analysis) system has been developed to meet ATLAS production and analysis requirements for a data-driven workload management system capable of operating at LHC data processing scale. PanDA has performed well with high reliability and robustness during the two years of LHC data-taking, while being actively evolved to meet the rapidly changing requirements for analysis use cases. We will present an overview of system evolution including automatic rebrokerage and reattempt for analysis jobs, adaptation for the CernVM File System, support for the multi-cloud model through which Tier-2 sites act as members of multiple clouds, pledged resource management and preferential brokerage, and monitoring improvements. We will also describe results from the analysis of two years of PanDA usage statistics, current issues, and plans for the future.

  3. The coolest DA white dwarfs detected at soft X-ray wavelengths

    NASA Technical Reports Server (NTRS)

    Kidder, K. M.; Holberg, J. B.; Barstow, M. A.; Tweedy, R. W.; Wesemael, F.

    1992-01-01

    New soft X-ray/EUV photometric observations of the DA white dwarfs KPD 0631 + 1043 = WD 0631 + 107 and PG 1113 + 413 = WD 1113 + 413 are analyzed. Previously reported soft X-ray detections of three other DAs and the failure to detect a fourth DA in deep Exosat observations are investigated. New ground-based spectra are presented for all of the objects, with IUE Ly-alpha spectra for some. These data are used to constrain the effective temperatures and surface gravities. The improved estimates of these parameters are employed to refer a photospheric He abundance for the hotter objects and to elucidate an effective observational low-temperature threshold for the detection of pure hydrogen DA white dwarfs at soft X-ray wavelengths.

  4. The coolest DA white dwarfs detected at soft X-ray wavelengths

    NASA Technical Reports Server (NTRS)

    Kidder, K. M.; Holberg, J. B.; Barstow, M. A.; Tweedy, R. W.; Wesemael, F.

    1992-01-01

    New soft X-ray/EUV photometric observations of the DA white dwarfs KPD 0631 + 1043 = WD 0631 + 107 and PG 1113 + 413 = WD 1113 + 413 are analyzed. Previously reported soft X-ray detections of three other DAs and the failure to detect a fourth DA in deep Exosat observations are investigated. New ground-based spectra are presented for all of the objects, with IUE Ly-alpha spectra for some. These data are used to constrain the effective temperatures and surface gravities. The improved estimates of these parameters are employed to refer a photospheric He abundance for the hotter objects and to elucidate an effective observational low-temperature threshold for the detection of pure hydrogen DA white dwarfs at soft X-ray wavelengths.

  5. Goldstone Radar Images of Near-Earth Asteroid 2012 DA14

    NASA Astrophysics Data System (ADS)

    Benner, Lance A.; Brozovic, M.; Giorgini, J. D.; Jao, J. S.; Lee, C. G.; Busch, M. W.; Slade, M. A.

    2013-10-01

    2012 DA14 made an extremely close approach within 0.00023 AU (0.09 lunar distances) on February 15, 2013. This was the closest encounter known in advance by a near-Earth asteroid with an absolute magnitude brighter than 25. We took advantage of this opportunity to image 2012 DA14 at Goldstone on February 16, 18, and 19 as the asteroid was receding from Earth. The imaging started at 00:46 UTC, slightly more than five hours after the closest approach, when the asteroid was at a distance of 0.00088 AU (0.34 lunar distances). The echoes were extraordinarily strong and we obtained some of the highest signal-to-noise ratios for any near-Earth asteroid ever observed at Goldstone. The images achieved resolutions as fine as 3.75 m x 0.00625 Hz that spatially resolve 2012 DA14 into a few dozen pixels; however, due to the diminutive dimensions of the asteroid, even this extremely high resolution is insufficient to show detailed surface features. The images indicate that 2012 DA14 is an asymmetric, angular, and elongated object with major axes of roughly 40 x 20 meters. The sequence of images obtained on February 16 spans 7.8 hours, during which the orientation of the asteroid appeared to complete close to one full rotation, suggesting that the rotation period is somewhat longer than ~8 hours. Radar speckle tracking with the Very Large Array and elements of the Very Long Baseline array show that 2012 DA14 has a relatively low obliquity and constrain the amplitude of any non-principal axis wobble after the flyby to be < 30 degrees. 2012 DA14 is among the smallest near-Earth asteroids ever spatially resolved by radar and it highlights the substantial scientific potential of the new chirp imaging system at Goldstone.

  6. Uppermost mantle and crustal structure at Tristan da Cunha derived from ambient seismic noise

    NASA Astrophysics Data System (ADS)

    Ryberg, T.; Geissler, W. H.; Jokat, W.; Pandey, S.

    2017-08-01

    According to classical plume theory, the Tristan da Cunha hotspot, located ∼400 km east off the Mid-Atlantic Ridge, is thought to have played a major role in the rifting while creating an aseismic Walvis Ridge during and after the breakup of the South Atlantic margins. Volcanic activity on the Tristan da Cunha Island shows that the hotspot might still be there influencing the upper mantle and crustal structure. In this study we present ambient noise data from 24 broadband OBS around Tristan da Cunha and a seismic station on Nightingale Island, which provide first constraints on the crustal and uppermost mantle structure around the island. By combining ambient noise techniques, dispersion curve analysis of Rayleigh waves, 2D tomographic inversion of travel times and 3D depth inversion of dispersion data we derived a 3D VS velocity model around the archipelago of Tristan da Cunha. The model shows an isolated, vertically sharp bounded thickened and modified crust beneath the islands surrounded by thin oceanic crust (<5 km). The velocity anomaly shows a typical volcanic structure with a shallow high velocity body and a low velocity root reaching into the upper mantle, which we interpret as shallow solidified magmatic material and volcanic feeding system/magmatic underplating, respectively. The observed simple and localized volcanic structure, embedded in a rather homogeneous crust and upper mantle indicates only minor and very localized magmatic overprinting of the existing lithosphere by the Tristan da Cunha hotspot. The uppermost mantle S wave velocity beneath nearby seamounts and to the SW of the islands is also slow and could indicate a thermal influence from a deeper source, whereas the Tristan da Cunha Fracture Zone shows no signs of modification.

  7. An analysis of DA white dwarfs from the Hamburg Quasar Survey

    NASA Astrophysics Data System (ADS)

    Homeier, D.; Koester, D.; Hagen, H.-J.; Jordan, S.; Heber, U.; Engels, D.; Reimers, D.; Dreizler, S.

    1998-10-01

    Follow-up spectroscopy of several hundred hot stars detected by the Hamburg Quasar Survey (HQS) has been carried out between 1989 and 1996. We present the analysis of 80 DA white dwarfs using model atmospheres and theoretical cooling tracks to derive the atmospheric parameters T_eff and log g, masses and absolute magnitudes. The HQS turned out to be sensitive to the detection of hydrogen-rich white dwarfs in a wide temperature range, from 10 000 K upwards. Star counts within four HQS fields for magnitudes B<= 16fm 4 exceed those from the Palomar Green survey by about 50%. The more recent observation campaigns emphasized the detection of very hot degenerates, yielding a large fraction of DA stars with T_eff > 50 000 K compared to other surveys. The mean mass of our DA sample is M=0.61{M}_{\\odot}, with three massive DA stars exceeding 1{M}_{\\odot} and three DA stars with masses significantly below the assummed lower mass limit for single white dwarf evolution of 0.45{M}_{\\odot}. Among the cool DA stars, thirteen are potential ZZ Ceti candidates because their effective temperatures lie close to the instability strip. Based on observations collected at the German-Spanish Astronomical Center (DSAZ), Calar Alto, operated by the Max-Plank-Institut für Astronomie Heidelberg jointly with the Spanish National Commission for Astronomy; with the International Ultraviolet Explorer satellite (IUE) collected at Villafranca, Spain, and with the ROSAT X-ray telescope.

  8. Integration of PanDA workload management system with Titan supercomputer at OLCF

    NASA Astrophysics Data System (ADS)

    De, K.; Klimentov, A.; Oleynik, D.; Panitkin, S.; Petrosyan, A.; Schovancova, J.; Vaniachine, A.; Wenaus, T.

    2015-12-01

    The PanDA (Production and Distributed Analysis) workload management system (WMS) was developed to meet the scale and complexity of LHC distributed computing for the ATLAS experiment. While PanDA currently distributes jobs to more than 100,000 cores at well over 100 Grid sites, the future LHC data taking runs will require more resources than Grid computing can possibly provide. To alleviate these challenges, ATLAS is engaged in an ambitious program to expand the current computing model to include additional resources such as the opportunistic use of supercomputers. We will describe a project aimed at integration of PanDA WMS with Titan supercomputer at Oak Ridge Leadership Computing Facility (OLCF). The current approach utilizes a modified PanDA pilot framework for job submission to Titan's batch queues and local data management, with light-weight MPI wrappers to run single threaded workloads in parallel on Titan's multicore worker nodes. It also gives PanDA new capability to collect, in real time, information about unused worker nodes on Titan, which allows precise definition of the size and duration of jobs submitted to Titan according to available free resources. This capability significantly reduces PanDA job wait time while improving Titan's utilization efficiency. This implementation was tested with a variety of Monte-Carlo workloads on Titan and is being tested on several other supercomputing platforms. Notice: This manuscript has been authored, by employees of Brookhaven Science Associates, LLC under Contract No. DE-AC02-98CH10886 with the U.S. Department of Energy. The publisher by accepting the manuscript for publication acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes.

  9. Pharmacokinetics of chlorogenic acid and corydaline in DA-9701, a new botanical gastroprokinetic agent, in rats.

    PubMed

    Jung, Ji Won; Kim, Ju Myung; Jeong, Jin Seok; Son, Miwon; Lee, Hye Suk; Lee, Myung Gull; Kang, Hee Eun

    2014-07-01

    1.Few studies describing the pharmacokinetic properties of chlorogenic acid (CA) and corydaline (CRD) which are marker compounds of a new prokinetic botanical agent, DA-9701, have been reported. The aim of the present study is to evaluate the pharmacokinetic properties CA and CRD following intravenous and oral administration of pure CA (1-8 mg/kg) or CRD (1.1-4.5 mg/kg) and their equivalent dose of DA-9701 to rats. 2.  Dose-proportional AUC and dose-independent clearance (10.3-12.1 ml/min/kg) of CA were observed following its administration. Oral administration of CA as DA-9701 did not influence the oral pharmacokinetic parameters of CA. Incomplete absorption of CA, its decomposition in the gastrointestinal tract, and/or pre-systemic metabolism resulted in extremely low oral bioavailability (F) of CA (0.478-0.899%). 3.  CRD showed greater dose-normalized AUC in the higher dose group than that in lower dose group(s) after its administration due to saturation of its metabolism via decreased non-renal clearance (by 51.3%) and first-pass extraction. As a result, the F of CRD following 4.5 mg/kg oral CRD (21.1%) was considerably greater than those of the lower dose groups (9.10 and 13.8%). However, oral administration of CRD as DA-9701 showed linear pharmacokinetics as a result of increased AUC and F in lower-dose groups (by 182% and 78.5%, respectively) compared to those of pure CRD. The greater oral AUC of CRD for DA-9701 than for pure CRD could be due to decreased hepatic and/or GI first-pass extraction of CRD by other components in DA-9701.

  10. Recurring X-ray outbursts in the supernova impostor SN 2010da in NGC 300

    NASA Astrophysics Data System (ADS)

    Binder, B.; Williams, B. F.; Kong, A. K. H.; Gaetz, T. J.; Plucinsky, P. P.; Skillman, E. D.; Dolphin, A.

    2016-04-01

    We present new observations of the `supernova impostor' SN 2010da using the Chandra X-ray Observatory and the Hubble Space Telescope. During the initial 2010 outburst, the 0.3-10 keV luminosity was observed by Swift to be ˜5 × 1038 erg s-1 and faded by a factor of ˜25 in a four month period. Our two new Chandra observations show a factor of ˜10 increase in the 0.35-8 keV X-ray luminosity, from ˜4 × 1036 to 4 × 1037 erg s-1 in ˜6 months, and the X-ray spectrum is consistent in both observations with a power-law with a photon index of Γ ˜ 0. We find evidence of X-ray spectral state changes: when SN 2010da is in a high-luminosity state, the X-ray spectrum is harder (Γ ˜0) compared to the low-luminosity state (Γ ˜ 1.2 ± 0.8). Using our Hubble observations, we fit the colour-magnitude diagram of the coeval stellar population to estimate a time since formation of the SN 2010da progenitor system of ≲5 Myr. Our observations are consistent with SN 2010da being a high-mass X-ray binary (HMXB) composed of a neutron star and a luminous blue variable-like companion, although we cannot rule out the possibility that SN 2010da is an unusually X-ray bright massive star. The ≲5 Myr age is consistent with the theoretically predicted delay time between the formation of a massive binary and the onset of the HMXB phase. It is possible that the initial 2010 outburst marked the beginning of X-ray production in the system, making SN 2010da possibly the first massive progenitor binary ever observed to evolve into an HMXB.

  11. Identification of an abundant 56 kDa protein implicated in food allergy as granule-bound starch synthase.

    PubMed

    Krishnan, Hari B; Chen, Ming-Hsuan

    2013-06-05

    Rice, the staple food of south and east Asian counties, is considered to be hypoallergenic. However, several clinical studies have documented rice-induced allergy in sensitive patients. Rice proteins with molecular weights of 14-16, 26, 33, and 56 kDa have been identified as allergens. Recently, it was documented that the 56 kDa rice allergen was responsible for rice-induced anaphylaxis. The 14-16 kDa allergens have been identified as α-amylase inhibitors; the 26 kDa protein has been identified as α-globulin; and the 33 kDa protein has been identified as glyoxalase I. However, the identity of the 56 kDa rice allergen has not yet been determined. In this study, we demonstrate that serum from patients allergic to maize shows IgE binding to a 56 kDa protein that was present in both maize and rice but not in the oil seeds soybean and peanut. The 56 kDa IgE-binding protein was abundant in the rice endosperm. We have purified this protein from rice endosperm and demonstrated its reactivity to IgE antibodies from the serum of maize-allergic patients. The purified protein was subjected to matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry analysis, resulting in identification of this rice allergen as granule-bound starch synthase, a product of the Waxy gene. Immunoblot analysis using protein extracts from a waxy mutant of rice revealed the absence of the 56 kDa IgE-binding protein. Our results demonstrate that the 56 kDa rice allergen is granule-bound starch synthase and raise the possibility of using waxy mutants of rice as a potential source of the hypoallergenic diet for patients sensitized to the 56 kDa rice allergen.

  12. Spectroscopy of the DA white dwarfs - Automatic atmospheric parameterization and mass distribution

    NASA Technical Reports Server (NTRS)

    Mcmahan, Robert K.

    1989-01-01

    A method for the automatic calculation of the atmospheric parameters (Teff and log g) of hydrogen-rich degenerate stars from low-resolution spectra is described, and then applied to the spectra of 53 DA white dwarfs. A value for the width of the DA mass distribution of sigma M/solar-M not greater than +0.10 is obtained using the proposed approach. The data indicate that the distribution is asymmetrically skewed to low masses; however, there is also evidence of a high-mass non-Gaussian tail.

  13. Evolution of robots throughout history from Hephaestus to Da Vinci Robot.

    PubMed

    Iavazzo, Christos; Gkegke, Xanthi-Ekaterini D; Iavazzo, Paraskevi-Evangelia; Gkegkes, Ioannis D

    2014-01-01

    Da Vinci robot is increasingly used for operations adding the advantages of robots to the favor of medicine. This is a historical article with the aim to present the evolution of robots in the medical area from the time of ancient myths to Renaissance and finally to the current revolutionary applications. We endeavored to collect several elegant narratives on the topic. The use of imagination could help the reader to find similarities. A trip from the Greek myths of Hephaestus through Aristotle and Leonardo Da Vinci to the robots of Karel Capek and Isaac Asimov and finally the invention of the medical robots is presented.

  14. Spectroscopy of the DA white dwarfs - Automatic atmospheric parameterization and mass distribution

    NASA Technical Reports Server (NTRS)

    Mcmahan, Robert K.

    1989-01-01

    A method for the automatic calculation of the atmospheric parameters (Teff and log g) of hydrogen-rich degenerate stars from low-resolution spectra is described, and then applied to the spectra of 53 DA white dwarfs. A value for the width of the DA mass distribution of sigma M/solar-M not greater than +0.10 is obtained using the proposed approach. The data indicate that the distribution is asymmetrically skewed to low masses; however, there is also evidence of a high-mass non-Gaussian tail.

  15. Solubilization of insoluble inorganic phosphate by Burkholderia cepacia DA23 isolated from cultivated soil

    PubMed Central

    Song, Ok-Ryul; Lee, Seung-Jin; Lee, Yong-Seok; Lee, Sang-Cheol; Kim, Keun-Ki; Choi, Yong-Lark

    2008-01-01

    A mineral phosphate solubilizing bacterium, Burkholderia cepacia DA23 has been isolated from cultivated soils. Phosphate-solubilizing activities of the strain against three types of insoluble phosphate were quantitatively determined. When 3% of glucose concentration was used for carbon source, the strain had a marked mineral phosphate-solubilizing activity. Mineral phosphate solubilization was directly related to the pH drop by the strain. Analysis of the culture medium by high pressure liquid chromatography identified gluconic acid as the main organic acid released by Burkholderia cepacia DA23. Gluconic acid production was apparently the result of the glucose dehydrogenase activity and glucose dehydrogenase was affected by phosphate regulation. PMID:24031195

  16. The First Korean Experience of Telemanipulative Robot-Assisted Laparoscopic Cholecystectomy Using the da Vinci System

    PubMed Central

    Kang, Chang Moo; Chi, Hoon Sang; Hyeung, Woo Jin; Kim, Kyung Sik; Choi, Jin Sub; Kim, Byong Ro

    2007-01-01

    With the advancement of laparoscopic instruments and computer sciences, complex surgical procedures are expected to be safely performed by robot assisted telemanipulative laparoscopic surgery. The da Vinci system (Intuitive Surgical, Mountain View, CA, USA) became available at the many surgical fields. The wrist like movements of the instrument's tip, as well as 3-dimensional vision, could be expected to facilitate more complex laparoscopic procedure. Here, we present the first Korean experience of da Vinci robotic assisted laparoscopic cholecystectomy and discuss the introduction and perspectives of this robotic system. PMID:17594166

  17. Complete and readily reversible blocking of striatal DaTscan binding by methylphenidate.

    PubMed

    Cheng, Gang; Morley, James F

    2014-02-01

    An 80-year-old man with Parkinson disease underwent DaTscan imaging, which showed no detectable uptake in the basal ganglia but normal background uptake in other parts of the brain and in the chest and abdomen. It was later found that the patient was taking methylphenidate. The second study performed 3 weeks later after discontinuing methylphenidate for 48 hours showed good uptake in the striatum. Our results indicate that methylphenidate may completely block physiologic uptake of DaTscan in the striatum and that this effect can be effectively reversed after discontinuing the medication for a short period.

  18. Direct Localization of the 51 kDa and 24 kDa Subunits of Mitochondrial Complex I by Three-dimensional Difference Imaging

    PubMed Central

    Clason, Todd; Zickermann, Volker; Ruiz, Teresa; Brandt, Ulrich; Radermacher, Michael

    2009-01-01

    Complex I is the largest complex in the respiratory chain, and the least understood. We have determined the three-dimensional structure of complex I from Yarrowia lipolytica lacking the flavoprotein part of the N-module, which consists of the 51 kDa (NUBM) and the 24 kDa (NUHM) subunits. The reconstruction was determined by three-dimensional electron microscopy of single particles. A comparison to our earlier reconstruction of the complete Y. lipolytica complex I clearly assigns the two flavoprotein subunits to an outer lobe of the peripheral arm of complex I. Localizing the two subunits allowed us to fit the X-ray structure of the hydrophilic fragment of complex I from Thermus thermophilus. The fit that is most consistent with previous immuno-electron microscopic data predicts that the ubiquinone reducing catalytic center resides in the second peripheral lobe, while the 75 kDa subunit is placed near the previously seen connection between the peripheral arm and the membrane arm protrusions. PMID:17591445

  19. The Settings, Pros and Cons of the New Surgical Robot da Vinci Xi System for Transoral Robotic Surgery (TORS): A Comparison With the Popular da Vinci Si System.

    PubMed

    Kim, Da Hee; Kim, Hwan; Kwak, Sanghyun; Baek, Kwangha; Na, Gina; Kim, Ji Hoon; Kim, Se Heon

    2016-10-01

    The da Vinci system (da Vinci Surgical System; Intuitive Surgical Inc.) has rapidly developed in several years from the S system to the Si system and now the Xi System. To investigate the surgical feasibility and to provide workflow guidance for the newly released system, we used the new da Vinci Xi system for transoral robotic surgery (TORS) on a cadaveric specimen. Bilateral supraglottic partial laryngectomy, hypopharyngectomy, lateral oropharyngectomy, and base of the tongue resection were serially performed in search of the optimal procedures with the new system. The new surgical robotic system has been upgraded in all respects. The telescope and camera were incorporated into one system, with a digital end-mounted camera. Overhead boom rotation allows multiquadrant access without axis limitation, the arms are now thinner and longer with grabbing movements for easy adjustments. The patient clearance button dramatically reduces external collisions. The new surgical robotic system has been optimized for improved anatomic access, with better-equipped appurtenances. This cadaveric study of TORS offers guidance on the best protocol for surgical workflow with the new Xi system leading to improvements in the functional results of TORS.

  20. Análise da aplicação e dos resultados do modelo OPM3® para a área da saúde

    PubMed Central

    Augusto dos Santos, Luis; de Fátima Marin, Heimar

    2015-01-01

    Esta pesquisa procurou analisar se um modelo de questionário criado por uma comunidade internacional de gerenciamento de projetos e se é aplicavél a organizações voltadas a área de saúde. O modelo OPM3® (Organizational Project Management Maturity Model) foi criado para que organizações de qualquer área ou porte pudessem identificar a presença, ou ausência, de boas práticas de gerenciamento. O objetivo da aplicação desse modelo é avaliar sempre a organização e não o entrevistado. No presente artigo, são apresentados os resultados da aplicação desse modelo em uma organização que possuía produtos e serviços de tecnologia da informação aplicados à área de saúde. Este estudo verificou que o modelo é aplicável de forma rápida e que a organização analisada possuía um número expressivo de boas práticas. PMID:26924862