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Sample records for aleaciones al-6si-3cu-xmg t6

  1. Quench sensitivity of hot extruded 6061-T6 and 6069-T6 aluminum alloys

    SciTech Connect

    Bergsma, S C; Kassner, M E; Li, X; Rosen, R S

    2000-08-08

    The purpose of this study is to investigate the quench sensitivity of mechanical properties of hot extruded 6061 and 6069 aluminum alloys. The relationship between mechanical properties and quench delzty time at various temperatures between 200-500 C was determined. It was concluded that the 6069-T6 was somewhat more quench sensitive than 6061, which may be consistent with the composition difference.

  2. Stabilization of T-6 jet fuel by mixtures of antioxidants

    SciTech Connect

    Golubeva, I.A.; Gutnikova, L.P.; Popova, T.V.; Vishnyakova, T.P.

    1985-09-01

    Work has been reported on synergistic combinations of additives for the stabilization of jet fuels. This paper investigates combinations of the additive KF-1 with dilauryl thiodipropionate (DLTDP). Data on the effects of these additive combinations on the post-test optical density, acidity and peroxide number of T-6 fuel is presented. Attention is also given to combinations of antioxidants with organometallic compounds. The stabilization of T-6 fuel by combination of KF-1 with phenosansesters of 3,5-di-tert-butyl-4-hydroxyphenylpropionic acid and pentaerythritol, diethylene glycol, ethylene glycol, and thiodiethylene glycol is investigated.

  3. Performance Evaluation of the T6 Ion Engine

    NASA Technical Reports Server (NTRS)

    Snyder, John Steven; Goebel, Dan M.; Hofer, Richard R.; Polk, James E.; Wallace, Neil C.; Simpson, Huw

    2010-01-01

    The T6 ion engine is a 22-cm diameter, 4.5-kW Kaufman-type ion thruster produced by QinetiQ, Ltd., and is baselined for the European Space Agency BepiColombo mission to Mercury and is being qualified under ESA sponsorship for the extended range AlphaBus communications satellite platform. The heritage of the T6 includes the T5 ion thruster now successfully operating on the ESA GOCE spacecraft. As a part of the T6 development program, an engineering model thruster was subjected to a suite of performance tests and plume diagnostics at the Jet Propulsion Laboratory. The engine was mounted on a thrust stand and operated over its nominal throttle range of 2.5 to 4.5 kW. In addition to the typical electrical and flow measurements, an E x B mass analyzer, scanning Faraday probe, thrust vector probe, and several near-field probes were utilized. Thrust, beam divergence, double ion content, and thrust vector movement were all measured at four separate throttle points. The engine performance agreed well with published data on this thruster. At full power the T6 produced 143 mN of thrust at a specific impulse of 4120 seconds and an efficiency of 64%; optimization of the neutralizer for lower flow rates increased the specific impulse to 4300 seconds and the efficiency to nearly 66%. Measured beam divergence was less than, and double ion content was greater than, the ring-cusp-design NSTAR thruster that has flown on NASA missions. The measured thrust vector offset depended slightly on throttle level and was found to increase with time as the thruster approached thermal equilibrium.

  4. Quantitative image analysis of WE43-T6 cracking behavior

    NASA Astrophysics Data System (ADS)

    Ahmad, A.; Yahya, Z.

    2013-06-01

    Environment-assisted cracking of WE43 cast magnesium (4.2 wt.% Yt, 2.3 wt.% Nd, 0.7% Zr, 0.8% HRE) in the T6 peak-aged condition was induced in ambient air in notched specimens. The mechanism of fracture was studied using electron backscatter diffraction, serial sectioning and in situ observations of crack propagation. The intermetallic (rare earthed-enriched divorced intermetallic retained at grain boundaries and predominantly at triple points) material was found to play a significant role in initiating cracks which leads to failure of this material. Quantitative measurements were required for this project. The populations of the intermetallic and clusters of intermetallic particles were analyzed using image analysis of metallographic images. This is part of the work to generate a theoretical model of the effect of notch geometry on the static fatigue strength of this material.

  5. A visual assay to monitor T6SS-mediated bacterial competition.

    PubMed

    Hachani, Abderrahman; Lossi, Nadine S; Filloux, Alain

    2013-03-20

    Type VI secretion systems (T6SSs) are molecular nanomachines allowing Gram-negative bacteria to transport and inject proteins into a wide variety of target cells(1,2). The T6SS is composed of 13 core components and displays structural similarities with the tail-tube of bacteriophages(3). The phage uses a tube and a puncturing device to penetrate the cell envelope of target bacteria and inject DNA. It is proposed that the T6SS is an inverted bacteriophage device creating a specific path in the bacterial cell envelope to drive effectors and toxins to the surface. The process could be taken further and the T6SS device could perforate other cells with which the bacterium is in contact, thus injecting the effectors into these targets. The tail tube and puncturing device parts of the T6SS are made with Hcp and VgrG proteins, respectively(4,5). The versatility of the T6SS has been demonstrated through studies using various bacterial pathogens. The Vibrio cholerae T6SS can remodel the cytoskeleton of eukaryotic host cells by injecting an "evolved" VgrG carrying a C-terminal actin cross-linking domain(6,7). Another striking example was recently documented using Pseudomonas aeruginosa which is able to target and kill bacteria in a T6SS-dependent manner, therefore promoting the establishment of bacteria in specific microbial niches and competitive environment(8,9,10). In the latter case, three T6SS-secreted proteins, namely Tse1, Tse2 and Tse3 have been identified as the toxins injected in the target bacteria (Figure 1). The donor cell is protected from the deleterious effect of these effectors via an anti-toxin mechanism, mediated by the Tsi1, Tsi2 and Tsi3 immunity proteins(8,9,10). This antimicrobial activity can be monitored when T6SS-proficient bacteria are co-cultivated on solid surfaces in competition with other bacterial species or with T6SS-inactive bacteria of the same species(8,11,12,13). The data available emphasized a numerical approach to the bacterial

  6. Atomic Structure and Mutagenesis of T6SS Reveals Interlaced Array Essential to Function

    PubMed Central

    Lee, Bai-Yu; Horwitz, Marcus A.; Zhou, Z. Hong

    2015-01-01

    Summary Type VI secretion systems (T6SSs) are newly identified contractile nanomachines that translocate effector proteins across bacterial membranes. The Francisella pathogenicity island, required for bacterial phagosome escape, intracellular replication and virulence, was presumed to encode a T6SS-like apparatus. Here, we experimentally confirm the identity of this T6SS and, by cryo electron microscopy (cryoEM), show the structure of its post-contraction sheath at 3.7 Å resolution. We demonstrate the assembly of this T6SS by IglA/IglB and secretion of its putative effector proteins in response to environmental stimuli. The sheath has a quaternary structure with handedness opposite that of contracted sheath of T4 phage tail and is organized in an interlaced two-dimensional array by means of β sheet augmentation. By structure-based mutagenesis, we show that this interlacing is essential to secretion, phagosomal escape, and intracellular replication. Our atomic model of the T6SS will facilitate design of drugs targeting this highly prevalent secretion apparatus. PMID:25723168

  7. Differential Role of the T6SS in Acinetobacter baumannii Virulence

    PubMed Central

    Foucault-Grunenwald, Marie-Laure; Borges, Vitor; Charpentier, Xavier; Limansky, Adriana S.; Gomes, João Paulo; Viale, Alejandro M.; Salcedo, Suzana P.

    2015-01-01

    Gram-negative bacteria, such as Acinetobacter baumannii, are an increasing burden in hospitals worldwide with an alarming spread of multi-drug resistant (MDR) strains. Herein, we compared a type strain (ATCC17978), a non-clinical isolate (DSM30011) and MDR strains of A. baumannii implicated in hospital outbreaks (Ab242, Ab244 and Ab825), revealing distinct patterns of type VI secretion system (T6SS) functionality. The T6SS genomic locus is present and was actively transcribed in all of the above strains. However, only the A. baumannii DSM30011 strain was capable of killing Escherichia coli in a T6SS-dependent manner, unlike the clinical isolates, which failed to display an active T6SS in vitro. In addition, DSM30011 was able to outcompete ATCC17978 as well as Pseudomonas aeruginosa and Klebsiella pneumoniae, bacterial pathogens relevant in mixed nosocomial infections. Finally, we found that the T6SS of DSM30011 is required for host colonization of the model organism Galleria mellonella suggesting that this system could play an important role in A. baumannii virulence in a strain-specific manner. PMID:26401654

  8. High strength and high ductility behavior of 6061-T6 alloy after laser shock processing

    NASA Astrophysics Data System (ADS)

    Gencalp Irizalp, Simge; Saklakoglu, Nursen

    2016-02-01

    The plastic deformation behavior of 6061-T6 alloy which was subjected to severe plastic deformation (SPD) at high strain rates during laser shock processing (LSP) was researched. In LSP-treated materials, the near surface microstructural change was examined by TEM and fracture surfaces after tensile testing were examined by SEM. An increase in strength of metallic materials brings about the decrease in ductility. In this study, the results showed that LSP-treated 6061-T6 alloy exhibited both high strength and high ductility. TEM observation showed that stacking fault (SF) ribbon enlarged, deformation twins formed and twin boundary increased in LSP-treated 6061-T6 alloy. This observation was an indication of stacking fault energy (SFE) decrease. Work hardening capability was recovered after LSP impacts.

  9. Numerical investigation on friction welding of alumina / AA 6063 T6 joints

    NASA Astrophysics Data System (ADS)

    Hynes, N. Rajesh Jesudoss; Velu, P. Shenbaga; Tharmaraj, R.; Kumar, R.

    2016-05-01

    Friction welding is the most suitable candidate for joining the dissimilar materials such as AA 6063 T6 alloy/Alumina for critical situations in high temperature and high corrosion resistance applications. In the present work, the numerical simulation was carried out to understand the mechanism of joining of AA 6063 T6 alloy/ Alumina rod by friction welding process. The developed thermo mechanical model is a highly nonlinear due to the interaction between the temperature fields and time dependent of the material properties. The developed numerical tool could be used to predict the temperature distribution, stress, strain and deformation of the dissimilar joints.

  10. Parametric studies on tensile strength in joining AA6061- T6 and AA7075-T6 by gas metal arc welding process

    NASA Astrophysics Data System (ADS)

    Ishak, M.; Noordin, N. F. M.; Shah, L. H.

    2015-12-01

    Proper selection of the welding parameters can result in better joining. In this study, the effects of various welding parameters on tensile strength in joining dissimilar aluminum alloys AA6061-T6 and AA7075-T6 were investigated. 2 mm thick samples of both base metals were welded by semi-automatic gas metal arc welding (GMAW) using filler wire ER5356. The welding current, arc voltage and welding speed were chosen as variables parameters. The strength of each specimen after the welding operations were tested and the effects of these parameters on tensile strength were identified by using Taguchi method. The range of parameter for welding current were chosen from 100 to 115 A, arc voltage from 17 to 20 V and welding speed from 2 to 5 mm/s. L16 orthogonal array was used to obtained 16 runs of experiments. It was found that the highest tensile strength (194.34 MPa) was obtained with the combination of a welding current of 115 A, welding voltage of 18 V and welding speed of 4 mm/s. Through analysis of variance (ANOVA), the welding voltage was the most effected parameter on tensile strength with percentage of contribution at 41.30%.

  11. Cavitation pitting and erosion of Al 6061-T6 in mineral oil and water

    NASA Technical Reports Server (NTRS)

    Rao, B. C. S.; Buckley, D. H.

    1983-01-01

    The authors are currently carrying out a study of the cavitation erosion of different bearing metals and alloys in mineral oils were studied. The variations of weight loss, the pit diameter and depth due to cavitation erosion on Al 6061-T6 in mineral oil and water are presented.

  12. Involvement of Trichoderma asperellum strain T6 in regulating iron acquisition in plants.

    PubMed

    Zhao, Lei; Wang, Fei; Zhang, Yaqing; Zhang, Jiaojiao

    2014-07-01

    Iron (Fe) deficiency is a major plant nutritional disorder in many parts of the world, particularly in areas with saline soils. Among the numerous root-associated microbes that are beneficial for plant nutrient uptake, Trichoderma spp. are the most effective rhizosphere fungi for enhancing plant growth and plant resistance to biotic and abiotic stresses. To investigate the potential mechanisms of action of Trichoderma on insoluble Fe in the soil, which is difficult for plants to absorb and utilize, a high siderophore-producing strain of Trichoderma T6, was isolated from the rhizosphere of cucumber plants. The strain was identified as T. asperellum based on the morphological features and molecular phylogenetic analyses. Applying strain T6 to sterile soil could increase soil levels of Fe(2+) and siderophores, as well as increase Fe(2+) and Fe(3+)-chelate reductase (FCR) activity in cucumber tissues. Purified siderophore eluent (PSE) increased plant growth, thus confirming its role in plant growth promotion. Moreover, extracellular Fe(3+) reducing activity and three kinds of organic acids were detected in the culture filtrate of strain T6. These results indicate that strain T6 influences plant Fe absorption in several ways. Siderophore-based Fe chelation is effective in providing Fe to plants, organic acids, and Fe(3+) reducing enzymes may participate in the solubilization and reduction of insoluble Fe(3+) to Fe(2+).

  13. Biosynthesis of Threonylcarbamoyl Adenosine (t6A), a Universal tRNA Nucleoside*

    PubMed Central

    Deutsch, Christopher; El Yacoubi, Basma; de Crécy-Lagard, Valérie; Iwata-Reuyl, Dirk

    2012-01-01

    The anticodon stem-loop (ASL) of transfer RNAs (tRNAs) drives decoding by interacting directly with the mRNA through codon/anticodon pairing. Chemically complex nucleoside modifications found in the ASL at positions 34 or 37 are known to be required for accurate decoding. Although over 100 distinct modifications have been structurally characterized in tRNAs, only a few are universally conserved, among them threonylcarbamoyl adenosine (t6A), found at position 37 in the anticodon loop of a subset of tRNA. Structural studies predict an important role for t6A in translational fidelity, and in vivo work supports this prediction. Although pioneering work in the 1970s identified the fundamental substrates for t6A biosynthesis, the enzymes responsible for its biosynthesis have remained an enigma. We report here the discovery that in bacteria four proteins (YgjD, YrdC, YjeE, and YeaZ) are both necessary and sufficient for t6A biosynthesis in vitro. Notably, YrdC and YgjD are members of universally conserved families that were ranked among the top 10 proteins of unknown function in need of functional characterization, while YeaZ and YjeE are specific to bacteria. This latter observation, coupled with the essentiality of all four proteins in bacteria, establishes this pathway as a compelling new target for antimicrobial development. PMID:22378793

  14. The T6SSs of Pseudomonas aeruginosa Strain PAO1 and Their Effectors: Beyond Bacterial-Cell Targeting

    PubMed Central

    Sana, Thibault G.; Berni, Benjamin; Bleves, Sophie

    2016-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen responsible for many diseases such as chronic lung colonization in cystic fibrosis patients and acute infections in hospitals. The capacity of P. aeruginosa to be pathogenic toward several hosts is notably due to different secretion systems. Amongst them, P. aeruginosa encodes three Type Six Secretion Systems (T6SS), named H1- to H3-T6SS, that act against either prokaryotes and/or eukaryotic cells. They are independent from each other and inject diverse toxins that interact with different components in the host cell. Here we summarize the roles of these T6SSs in the PAO1 strain, as well as the toxins injected and their targets. While H1-T6SS is only involved in antiprokaryotic activity through at least seven different toxins, H2-T6SS and H3-T6SS are also able to target prokaryotic as well as eukaryotic cells. Moreover, recent studies proposed that H2- and H3-T6SS have a role in epithelial cells invasion by injecting at least three different toxins. The diversity of T6SS effectors is astounding and other effectors still remain to be discovered. In this review, we present a table with other putative P. aeruginosa strain PAO1 T6SS-dependent effectors. Altogether, the T6SSs of P. aeruginosa are important systems that help fight other bacteria for their ecological niche, and are important in the pathogenicity process. PMID:27376031

  15. The T6SSs of Pseudomonas aeruginosa Strain PAO1 and Their Effectors: Beyond Bacterial-Cell Targeting.

    PubMed

    Sana, Thibault G; Berni, Benjamin; Bleves, Sophie

    2016-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen responsible for many diseases such as chronic lung colonization in cystic fibrosis patients and acute infections in hospitals. The capacity of P. aeruginosa to be pathogenic toward several hosts is notably due to different secretion systems. Amongst them, P. aeruginosa encodes three Type Six Secretion Systems (T6SS), named H1- to H3-T6SS, that act against either prokaryotes and/or eukaryotic cells. They are independent from each other and inject diverse toxins that interact with different components in the host cell. Here we summarize the roles of these T6SSs in the PAO1 strain, as well as the toxins injected and their targets. While H1-T6SS is only involved in antiprokaryotic activity through at least seven different toxins, H2-T6SS and H3-T6SS are also able to target prokaryotic as well as eukaryotic cells. Moreover, recent studies proposed that H2- and H3-T6SS have a role in epithelial cells invasion by injecting at least three different toxins. The diversity of T6SS effectors is astounding and other effectors still remain to be discovered. In this review, we present a table with other putative P. aeruginosa strain PAO1 T6SS-dependent effectors. Altogether, the T6SSs of P. aeruginosa are important systems that help fight other bacteria for their ecological niche, and are important in the pathogenicity process. PMID:27376031

  16. Cleidocranial dysplasia associated with a t(6;18)(p12;q24) translocation

    SciTech Connect

    Narahara, Kouji; Tsuji, Kazushiro; Yokoyama, Yuji

    1995-03-13

    Cleidocranial displasia (CCD) is an autosomal-dominant, generalized skeletal disorder. The gene locus for this syndrome has not yet been mapped. The presence of rare chromosome rearrangements in Mendelian disorders has facilitated the localization or positional cloning of these disease genes. We report on another patient with autosomal-dominant CCD who had a balanced t(6;8) translocation. 3 refs., 2 figs.

  17. Purification and characterization of alpha-L-arabinofuranosidase from Bacillus stearothermophilus T-6.

    PubMed Central

    Gilead, S; Shoham, Y

    1995-01-01

    Bacillus stearothermophilus T-6 produced an alpha-L-arabinofuranosidase when grown in the presence of L-arabinose, sugar beet arabinan, or oat spelt xylan. At the end of a fermentation, about 40% of the activity was extracellular, and enzyme activity in the cell-free supernatant could reach 25 U/ml. The enzymatic activity in the supernatant was concentrated against polyethylene glycol 20000, and the enzyme was purified eightfold by anion-exchange and hydrophobic interaction chromatographies. The molecular weight of T-6 alpha-L-arabinofuranosidase was 256,000, and it consisted of four identical subunits as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration. The native enzyme had a pI of 6.5 and was most active at 70 degrees C and at pH 5.5 to 6.0. Its thermostability at pH 7.0 was characterized by half-lives of 53, 15, and 1 h at 60, 65, and 70 degrees C, respectively. Kinetic experiments at 60 degrees C with p-nitrophenyl alpha-L-arabinofuranoside as a substrate gave a Vmax, a Km, and an activation energy of 749 U/mg, 0.42 mM, and 16.6 kcal/mol, (ca. 69.5 kJ/mol), respectively. The enzyme had no apparent requirement for cofactors, and its activity was strongly inhibited by 1 mM Hg2+. T-6 alpha-L-arabinofuranosidase released L-arabinose from arabinan and had low activity on oat spelt xylan. The enzyme acted cooperatively with T-6 xylanase in hydrolyzing oat spelt xylan, and L-arabinose, xylose, and xylobiose were detected as the end reaction products.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7887599

  18. Functional performance of the T-6A Texan (JPATS) CFIS laser detonator

    NASA Astrophysics Data System (ADS)

    Blachowski, Thomas J.; Thom, Travis

    2010-08-01

    The Indian Head Division, Naval Surface Warfare Center (IHD NSWC) CAD Engineering Division is conducting a program to evaluate the laser and energetic components which comprise the Canopy Fracturing Initiation System (CFIS). This system is currently installed on the T-6A Texan II or JPATS (Joint Primary Aircraft Training System) aircraft. The T-6A Texan II is the first aircraft used by the military to train future pilots. The CFIS is an element of the pilot emergency escape system which weakens the canopy in the path of the ejection seat. The CFIS is comprised of three differing laser configurations (Internal, External, and Seat Motion) which generate a pulse that is distributed through a fiber optic energy transmission system. This pulse, in turn, initiates one of the system's explosive components, a detonator (specifically, the CCU-158/A Laser Initiated Detonator). This detonator transfers the signal to the remaining energetic components that, in turn, function to weaken their respective canopies. All of the CFIS laser types are flashlamp-pumped, neodymium glass lasers which are located at various positions in the aircraft cockpit area. This paper builds on the previous SPIE papers (2008 - Conference 7070 and 2009 - Conference 7434, respectively) and presents the initial functional test results for the CFIS Laser Detonator. These functional test results provide the technical support to justify the useful lifetime of this energetic component while being installed in the T-6A Texan II aircraft under operational conditions.

  19. Effects of different ingredients of zedoary on gene expression of HSC-T6 cells

    PubMed Central

    Jiang, Yuan; Li, Ze-Song; Jiang, Fu-Sheng; Deng, Xin; Yao, Cong-Shun; Nie, Guang

    2005-01-01

    AIM: To investigate the effects of four different ingredients of zedoary (Curcuma aromatica oil, Curcumol, β-elemence, and Curcumin) on the gene expressions of hepatic stellate cells (HSCs), and to explore the molecular mechanism of zedoary against hepatic fibrosis at gene network level. METHODS: We detected the mRNA sequences of 50 liver fibrosis-related genes in GenBank and designed oligonucleotide probes. We synthesized oligonucleotides with PE8909 DNA synthesizing instrument, and carried out oligonucleotide microarray with OGR-04 dropping instrument and aldehyded glass chip. Cultured HSC-T6 cells were treated with different concentrations of Colchicine, Curcuma aromatica oil, Curcumol, β-elemence, and Curcumin. According to the experiment of cell toxicity, we took the appropriate concentrations of medicines that resulted in over 50% of cell survival as experiment concentrations. We collected the cells at 1, 6, 12, and 24 h, and extracted total RNA with TRIzol reagent, then labeled cDNAs with Cy3-dUTP and Cy5-dUTP. These labeled cDNAs were hybridized to an oligonucleotide microarray which was washed several times and scanned by scanner GenePix 4000B. Different gene expressions of HSC-T6 cells were analyzed by ImaGene 4.2 software. RESULTS: After HSC-T6 cells were cultured in a medium containing 6.25 μg/mL Colchicine for 12 h, expression of TIMP-1 decreased 2.2-folds. After HSC-T6 cells were cultured in a medium containing 78.125 μg/mL of Curcuma aromatica oil for 24 h, the expression of TIMP-2 and IL-6 decreased 2.3- and 2.2-folds, respectively. Moreover, after HSC-T6 cells were cultured in a medium containing 1.5625 μg/mL of Curcumol for 12 h, the expression of TGFβ1 and P450a decreased 2.3- and 2.1-folds, respectively. CONCLUSION: Our results may show the possible molecular mechanism of Curcuma aromatica oil and Curcumol against hepatic fibrosis. PMID:16425384

  20. Global translational impacts of the loss of the tRNA modification t6A in yeast

    PubMed Central

    Thiaville, Patrick C.; Legendre, Rachel; Rojas-Benítez, Diego; Baudin-Baillieu, Agnès; Hatin, Isabelle; Chalancon, Guilhem; Glavic, Alvaro; Namy, Olivier; de Crécy-Lagard, Valérie

    2016-01-01

    The universal tRNA modification t6A is found at position 37 of nearly all tRNAs decoding ANN codons. The absence of t6A37 leads to severe growth defects in baker’s yeast, phenotypes similar to those caused by defects in mcm5s2U34 synthesis. Mutants in mcm5s2U34 can be suppressed by overexpression of tRNALysUUU, but we show t6A phenotypes could not be suppressed by expressing any individual ANN decoding tRNA, and t6A and mcm5s2U are not determinants for each other’s formation. Our results suggest that t6A deficiency, like mcm5s2U deficiency, leads to protein folding defects, and show that the absence of t6A led to stress sensitivities (heat, ethanol, salt) and sensitivity to TOR pathway inhibitors. Additionally, L-homoserine suppressed the slow growth phenotype seen in t6A-deficient strains, and proteins aggregates and Advanced Glycation End-products (AGEs) were increased in the mutants. The global consequences on translation caused by t6A absence were examined by ribosome profiling. Interestingly, the absence of t6A did not lead to global translation defects, but did increase translation initiation at upstream non-AUG codons and increased frame-shifting in specific genes. Analysis of codon occupancy rates suggests that one of the major roles of t6A is to homogenize the process of elongation by slowing the elongation rate at codons decoded by high abundance tRNAs and I34:C3 pairs while increasing the elongation rate of rare tRNAs and G34:U3 pairs. This work reveals that the consequences of t6A absence are complex and multilayered and has set the stage to elucidate the molecular basis of the observed phenotypes. PMID:26798630

  1. Supernova Simulations from the T-6 Group at Los Alamos National Laboratory (LANL)

    DOE Data Explorer

    Woosley, Stanford

    LANL's primary effort to numerically model supernova explosions is based in the Theoretical Astrophysics Group (T-6). Both thermonuclear supernovae and core- collapse supernovae are studied, with special emphasis placed on multi-dimensional simulations. Both types of supernova require a wide range of input physics, which is provided by research efforts throughout the lab. In particular this research benefits from other LANL efforts studying massive star evolution, equations of state and aspects of neutrino physics. [From http://laastro.lanl.gov/science/computation.html

  2. An evaluation of the T-6A Texan (JPATS) functional performance of the CFIS laser assemblies

    NASA Astrophysics Data System (ADS)

    Blachowski, Thomas J.; Eccard, George

    2008-08-01

    The Indian Head Division, Naval Surface Warfare Center (IHDIV, NSWC) CAD Engineering Division is conducting a program to evaluate the laser components which comprise the Canopy Fracturing Initiation System (CFIS) currently installed on the T-6A Texan or JPATS (Joint Primary Aircraft Training System) aircraft. The T-6A Texan is the first aircraft used by the military to train future pilots. The CFIS is an element of the pilot emergency escape system which weakens the canopy in the path of the ejection seat. The CFIS is comprised of three differing configurations (Internal, External, and Seat Motion) which generate a laser pulse that is distributed through a fiber optic energy transmission system. This pulse, in turn, initiates explosive components which weaken the respective canopies. All of the CFIS laser types are flashlamp-pumped, neodymium glass lasers which are located at various positions in the aircraft cockpit area. This paper presents the CAD Engineering Division effort to evaluate the functional performance of the three CCFIS laser signal generators after their being in fleet use for a period of time.

  3. Differentiation of A31T6 proadipocytes to adipocytes: A flow cytometric analysis

    SciTech Connect

    Smyth, M.J.; Wharton, W. )

    1992-03-01

    A flow cytometric assay has been developed which provides precise, quantitative information on the accumulation of cytoplasmic triglycerides in individual A31T6 proadipocytes as they differentiate into adipocytes. The opportunity to measure multiple optical parameter on a cell-by-cell basis has enabled us to monitor phenotypic aspects of differentiation with a greater level of sensitivity than was previously possible. Using the fluorescent hydrophobic probe, Nile red, they have found that as a cell proceeds along the differentiation pathway, the gold fluorescence signal from the cell increases, reflecting the accumulation of cytoplasmic lipid droplets. They have determined (1) the presence of an undifferentiated population of cells whose existence is not detected by conventional phase microscopy, (2) that insulin is no required to drive differentiation in this system, (3) that exposure to a combination of insulin and dexamethasone results in a lower accumulation of lipid in a cell than does exposure to either agent alone, and (4) that A31T6 cells show the same response to differentiation-promoting agents whether applied at the time of plating or at confluence.

  4. Purification and characterization of a thermostable xylanase from Bacillus stearothermophilus T-6.

    PubMed Central

    Khasin, A; Alchanati, I; Shoham, Y

    1993-01-01

    Bacillus stearothermophilus T-6 produces an extracellular xylanase that was shown to optimally bleach pulp at pH 9 and 65 degrees C. The enzyme was purified and concentrated in a single adsorption step onto a cation exchanger and is made of a single polypeptide with an apparent M(r) of 43,000 (determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Xylanase T-6 is an endoxylanase that completely degrades xylan to xylose and xylobiose. The pIs of the purified protein were 9 and 7 under native and denaturing conditions, respectively. The optimum activity was at pH 6.5; however, 60% of the activity was still retained at pH 10. At 65 degrees C and pH 7, the enzyme was stable for more than 10 h; at 65 degrees C and pH 9, the half-life of the enzyme was approximately 6 h. Kinetic experiments at 55 degrees C gave Vmax and Km values of 288 U/mg and 1.63 mg/ml, respectively. The enzyme had no apparent requirement for cofactors, and its activity was strongly inhibited by Zn2+, Cd2+, and Hg2+. Xylan completely protected the protein from inactivation by N-bromosuccinimide. The N-terminal sequence of the first 45 amino acids of the enzyme showed high homology with the N-terminal region of xylanase A from the alkalophilic Bacillus sp. strain C-125. Images PMID:8328796

  5. Salmonella Typhimurium utilizes a T6SS-mediated antibacterial weapon to establish in the host gut.

    PubMed

    Sana, Thibault G; Flaugnatti, Nicolas; Lugo, Kyler A; Lam, Lilian H; Jacobson, Amanda; Baylot, Virginie; Durand, Eric; Journet, Laure; Cascales, Eric; Monack, Denise M

    2016-08-23

    The mammalian gastrointestinal tract is colonized by a high-density polymicrobial community where bacteria compete for niches and resources. One key competition strategy includes cell contact-dependent mechanisms of interbacterial antagonism, such as the type VI secretion system (T6SS), a multiprotein needle-like apparatus that injects effector proteins into prokaryotic and/or eukaryotic target cells. However, the contribution of T6SS antibacterial activity during pathogen invasion of the gut has not been demonstrated. We report that successful establishment in the gut by the enteropathogenic bacterium Salmonella enterica serovar Typhimurium requires a T6SS encoded within Salmonella pathogenicity island-6 (SPI-6). In an in vitro setting, we demonstrate that bile salts increase SPI-6 antibacterial activity and that S Typhimurium kills commensal bacteria in a T6SS-dependent manner. Furthermore, we provide evidence that one of the two T6SS nanotube subunits, Hcp1, is required for killing Klebsiella oxytoca in vitro and that this activity is mediated by the specific interaction of Hcp1 with the antibacterial amidase Tae4. Finally, we show that K. oxytoca is killed in the host gut in an Hcp1-dependent manner and that the T6SS antibacterial activity is essential for Salmonella to establish infection within the host gut. Our findings provide an example of pathogen T6SS-dependent killing of commensal bacteria as a mechanism to successfully colonize the host gut. PMID:27503894

  6. [Efficiency of Trichoderma longibrachiatum T6 in the control of Meloidogyne incognita and its rhizosphere colonization in cucumber].

    PubMed

    Zhang, Shu-wu; Xu, Bing-liang; Xue, Ying-yu; Liang, Qiao-lan; Liu, Jia

    2016-01-01

    Efficiency of different concentrations of Trichoderma longibrachiatum T6 against Meloidogyne incognita and its rhizosphere colonization in cucumber were determined in greenhouse experiments. The results of rhizosphere colonization experiments showed that the number of colonies in cucumber soil and root increased significantly ten weeks after inoculation with the second stage juveniles of M. incognita and different concentrations of T. longibrachiatum T6, and there was significant difference in different concentrations of T. longibrachiatum T6, e.g., the maximum numbers of colonies in soil and root were 7.8 x 10⁷ and 6.3 x 10⁵ CFU · mL⁻¹ respectively after treated with the spore suspension of 1.5 x 10⁷ CFU · mL⁻¹. Greenhouse experiments results showed that different concentrations of T. longibrachiatum T6 had significant control effect on different life stages of M. incognita, and the control effect increased with the concentration of T. longibrachiatum T6. T. longibrachiatum T6 significantly increased plant height, root length, above-ground and root fresh mass o cucumber inoculated by M. incognita. T. longibrachiatum T6 could colonize in cucumber rhizosphere, had control effect on M. incognita, and significantly improved the growth of cucumber.

  7. Salmonella Typhimurium utilizes a T6SS-mediated antibacterial weapon to establish in the host gut

    PubMed Central

    Flaugnatti, Nicolas; Lugo, Kyler A.; Lam, Lilian H.; Jacobson, Amanda; Baylot, Virginie; Durand, Eric; Journet, Laure; Cascales, Eric; Monack, Denise M.

    2016-01-01

    The mammalian gastrointestinal tract is colonized by a high-density polymicrobial community where bacteria compete for niches and resources. One key competition strategy includes cell contact-dependent mechanisms of interbacterial antagonism, such as the type VI secretion system (T6SS), a multiprotein needle-like apparatus that injects effector proteins into prokaryotic and/or eukaryotic target cells. However, the contribution of T6SS antibacterial activity during pathogen invasion of the gut has not been demonstrated. We report that successful establishment in the gut by the enteropathogenic bacterium Salmonella enterica serovar Typhimurium requires a T6SS encoded within Salmonella pathogenicity island-6 (SPI-6). In an in vitro setting, we demonstrate that bile salts increase SPI-6 antibacterial activity and that S. Typhimurium kills commensal bacteria in a T6SS-dependent manner. Furthermore, we provide evidence that one of the two T6SS nanotube subunits, Hcp1, is required for killing Klebsiella oxytoca in vitro and that this activity is mediated by the specific interaction of Hcp1 with the antibacterial amidase Tae4. Finally, we show that K. oxytoca is killed in the host gut in an Hcp1-dependent manner and that the T6SS antibacterial activity is essential for Salmonella to establish infection within the host gut. Our findings provide an example of pathogen T6SS-dependent killing of commensal bacteria as a mechanism to successfully colonize the host gut. PMID:27503894

  8. [Efficiency of Trichoderma longibrachiatum T6 in the control of Meloidogyne incognita and its rhizosphere colonization in cucumber].

    PubMed

    Zhang, Shu-wu; Xu, Bing-liang; Xue, Ying-yu; Liang, Qiao-lan; Liu, Jia

    2016-01-01

    Efficiency of different concentrations of Trichoderma longibrachiatum T6 against Meloidogyne incognita and its rhizosphere colonization in cucumber were determined in greenhouse experiments. The results of rhizosphere colonization experiments showed that the number of colonies in cucumber soil and root increased significantly ten weeks after inoculation with the second stage juveniles of M. incognita and different concentrations of T. longibrachiatum T6, and there was significant difference in different concentrations of T. longibrachiatum T6, e.g., the maximum numbers of colonies in soil and root were 7.8 x 10⁷ and 6.3 x 10⁵ CFU · mL⁻¹ respectively after treated with the spore suspension of 1.5 x 10⁷ CFU · mL⁻¹. Greenhouse experiments results showed that different concentrations of T. longibrachiatum T6 had significant control effect on different life stages of M. incognita, and the control effect increased with the concentration of T. longibrachiatum T6. T. longibrachiatum T6 significantly increased plant height, root length, above-ground and root fresh mass o cucumber inoculated by M. incognita. T. longibrachiatum T6 could colonize in cucumber rhizosphere, had control effect on M. incognita, and significantly improved the growth of cucumber. PMID:27228616

  9. T6BP and NDP52 are myosin VI binding partners with potential roles in cytokine signalling and cell adhesion

    PubMed Central

    Morriswood, Brooke; Ryzhakov, Grigory; Puri, Claudia; Arden, Susan D.; Roberts, Rhys; Dendrou, Calliope; Kendrick-Jones, John; Buss, Folma

    2009-01-01

    Summary Myosin VI has been implicated in many cellular processes including endocytosis, secretion, membrane ruffling and cell motility. We carried out a yeast two-hybrid screen and identified TRAF6-binding protein (T6BP) and nuclear dot protein 52 (NDP52) as myosin VI binding partners. Myosin VI interaction with T6BP and NDP52 was confirmed in vitro and in vivo and the binding sites on each protein were accurately mapped. Immunofluorescence and electron microscopy showed that T6BP, NDP52 and myosin VI are present at the trans side of the Golgi complex, and on vesicles in the perinuclear region. Although the SKICH domain in T6BP and NDP52 does not mediate recruitment into membrane ruffles, loss of T6BP and NDP52 in RNAi knockdown cells results in reduced membrane ruffling activity and increased stress fibre and focal adhesion formation. Furthermore, we observed in these knockdown cells an upregulation of constitutive secretion of alkaline phosphatase, implying that both proteins act as negative regulators of secretory traffic at the Golgi complex. T6BP was also found to inhibit NF-κB activation, implicating it in the regulation of TRAF6-mediated cytokine signalling. Thus myosin VI-T6BP interactions may link membrane trafficking pathways with cell adhesion and cytokine-dependent cell signalling. PMID:17635994

  10. Prevention of non-ductile fracture in 6061-T6 aluminum nuclear pressure vessels

    SciTech Connect

    Yahr, G.T.

    1995-06-01

    The American Society of Mechanical Engineers (ASME) Boiler and Pressure Vessel Committee has approved rules for the use of 6061-T6 and 6061-T651 aluminum for the construction of Class 1 welded nuclear pressure vessels for temperatures not exceeding 149 C (300 F). Nuclear Code Case N-519 allows the use of this aluminum in the construction of low temperature research reactors such as the Advanced Neutron Source. The rules for protection against non-ductile fracture are discussed. The basis for a value of 25.3 MPa {radical}m (23 ksi {radical}in.) for the critical or reference stress intensity factor for use in the fracture analysis is presented. Requirements for consideration of the effects of neutron irradiation on the fracture toughness are discussed.

  11. Built-up edge investigation in vibration drilling of Al2024-T6.

    PubMed

    Barani, A; Amini, S; Paktinat, H; Fadaei Tehrani, A

    2014-07-01

    Adding ultrasonic vibrations to drilling process results in an advanced hybrid machining process, entitled "vibration drilling". This study presents the design and fabrication of a vibration drilling tool by which both rotary and vibrating motions are applied to drill simultaneously. High frequency and low amplitude vibrations were generated by an ultrasonic transducer with frequency of 19.65 kHz. Ultrasonic transducer was controlled by a MPI ultrasonic generator with 3 kW power. The drilling tool and workpiece material were HSS two-flute twist drill and Al2024-T6, respectively. The aim of this study was investigating on the effect of ultrasonic vibrations on built-up edge, surface quality, chip morphology and wear mechanisms of drill edges. Therefore, these factors were studied in both vibration and ordinary drilling. Based on the achieved results, vibration drilling offers less built-up edge and better surface quality compared to ordinary drilling.

  12. A Micro-Electrochemical Study of Friction Stir Welded Aluminum 6061-T6

    NASA Technical Reports Server (NTRS)

    Hintze, Paul E.; Calle, Luz M.

    2005-01-01

    The corrosion behavior of friction stir welded Aluminum alloy 606 1-T6 was studied using a micro-electrochemical cell. The micro-electrochemical cell has a measurement area of about 0.25 square mm which allows for measurement of corrosion properties at a very small scale. The corrosion and breakdown potentials were measured at many points inside and outside the weld along lines perpendicular to the weld. The breakdown potential is approximately equal inside and outside the weld; however, it is lower in the narrow border between the weld and base material. The results of electrochemical measurements were correlated to micro-structural analysis. The corrosion behavior of the friction stir welded samples was compared to tungsten inert gas (TIG) welded samples of the same material.

  13. Built-up edge investigation in vibration drilling of Al2024-T6.

    PubMed

    Barani, A; Amini, S; Paktinat, H; Fadaei Tehrani, A

    2014-07-01

    Adding ultrasonic vibrations to drilling process results in an advanced hybrid machining process, entitled "vibration drilling". This study presents the design and fabrication of a vibration drilling tool by which both rotary and vibrating motions are applied to drill simultaneously. High frequency and low amplitude vibrations were generated by an ultrasonic transducer with frequency of 19.65 kHz. Ultrasonic transducer was controlled by a MPI ultrasonic generator with 3 kW power. The drilling tool and workpiece material were HSS two-flute twist drill and Al2024-T6, respectively. The aim of this study was investigating on the effect of ultrasonic vibrations on built-up edge, surface quality, chip morphology and wear mechanisms of drill edges. Therefore, these factors were studied in both vibration and ordinary drilling. Based on the achieved results, vibration drilling offers less built-up edge and better surface quality compared to ordinary drilling. PMID:24582556

  14. Cavitation pitting and erosion of aluminum 6061-T6 in mineral oil water

    NASA Technical Reports Server (NTRS)

    Rao, B. C. S.; Buckley, D. H.

    1983-01-01

    Cavitation erosion studies of aluminum 6061-T6 in mineral oil and in ordinary tap water are presented. The maximum erosion rate (MDPR, or mean depth of penetration rate) in mineral oil was about four times that in water. The MDPR in mineral oil decreased continuously with time, but the MDPR in water remained approximately constant. The cavitation pits in mineral oil were of smaller diameter and depth than the pits in water. Treating the pits as spherical segments, we computed the radius r of the sphere. The logarithm of h/a, where h is the pit depth and 2a is the top width of the pit, was linear when plotted against the logarithm of 2r/h - 1.

  15. The corrosion protection of 6061-T6 aluminum by a polyurethane-sealed anodized coat

    NASA Technical Reports Server (NTRS)

    Danford, M. D.

    1990-01-01

    The corrosion protection of 6061-T6 anodized aluminum afforded by a newly patented polyurethane seal was studied using the ac impedance technique. Values of the average corrosion rates over a 27-day exposure period in 3.5 percent NaCl solutions at pH 5.2 and pH 9.5 compared very favorably for Lockheed-prepared polyurethane-sealed and dichromate-sealed coats of the same thickness. Average corrosion rates for both specimens over the first 7 days of exposure compared well with those for a hard anodized, dichromate-sealed coat, but rose well above those for the hard anodized coat over the entire 27-day period. This is attributed both to the greater thickness of the hard anodized coat, and possibly to its inherently better corrosion protective capability.

  16. Effect of turning parameters on A16351 T6 by using Design of Experiments

    NASA Astrophysics Data System (ADS)

    Annigeri, Ulhas K.; Charan, M.; Vishnu Sai, M.; Ram Charan, T.; Rahul Sai, L.

    2016-09-01

    Aluminium is a well known lightest engineering metals. It is the most productive element on the earth crust. It can be extracted from bauxite, kaolinite or nepheline. It can be used in making aeroplane bodies, automobile parts etc. It is mostly used in multiple applications in order to reduce the weight of the component and it has fancy corrosion resistance. In the present study Al 6351 T6 is selected and a plain turning operation is performed on it with the help of design of experiments. The surface roughness of the samples are tested by varying the cutting parameters such as speed, feed, and depth of cut. A mathematical model is developed and the parameter which affects the surface roughness is determined.

  17. Effects of select parameters on electron beam welding of AL6061-T6 alloy

    NASA Astrophysics Data System (ADS)

    Yost, Thomas E.

    Electron beam welding was used for joining Al6061-T6, precision machined, cylindrical sections. The welded assembly exhibited a minimum amount of distortion, but a better understanding of the effects of several key welding parameters on the structural integrity of the weld was required. The contents of this document describe the relative importance and interaction between welding speed, volume of filler, and beam pattern on the microstructural and mechanical properties of the welded joint. Understanding of the relationship between welding parameters and weld properties was accomplished by macrophotography and microstructural examination, microhardness testing, energy dispersive spectroscopy (EDX), and mechanical tensile testing of weld coupons. The results of this study will help quantify the robustness of the EBW process for this common aerospace material and joint geometry and will help determine the impacts of process deviations on weld fidelity in the production environment.

  18. Sperm FISH and chromatin integrity in spermatozoa from a t(6;10;11) carrier.

    PubMed

    Olszewska, Marta; Huleyuk, Nataliya; Fraczek, Monika; Zastavna, Danuta; Wiland, Ewa; Kurpisz, Maciej

    2014-05-01

    Complex chromosome rearrangements (CCRs) are structurally balanced or unbalanced aberrations involving more than two breakpoints on two or more chromosomes. CCRs can be a potential reason for genomic imbalance in gametes, which leads to a drastic reduction in fertility. In this study, the meiotic segregation pattern, aneuploidy of seven chromosomes uninvolved in the CCR and chromatin integrity were analysed in the ejaculated spermatozoa of a 46,XY,t(6;10;11)(q25.1;q24.3;q23.1)mat carrier with asthenozoospermia and a lack of conception. The frequency of genetically unbalanced spermatozoa was 78.8% with a prevalence of 4:2 segregants of 38.2%, while the prevalence of the adjacent 3:3 mode was 35.3%. Analysis of the aneuploidy of chromosomes 13, 15, 18, 21, 22, X and Y revealed an approximately fivefold increased level in comparison with that of the control group, indicating the presence of an interchromosomal effect. Sperm chromatin integrity status was evaluated using chromomycin A3 and aniline blue staining (deprotamination), acridine orange test and TUNEL assay (sperm DNA fragmentation). No differences were found when comparisons were made with a control group. We suggest that the accumulation of genetically unbalanced spermatozoa, significantly increased sperm aneuploidy level and decreased sperm motility (20%, progressive) were not responsible for the observed lack of reproductive success in the analysed infertile t(6;10;11) carrier. Interestingly, in the case described herein, a high level of sperm chromosomal imbalance appears not to be linked to sperm chromatin integrity status.

  19. Early state damage detection of aluminum 7075-T6 plate based on acoustic emission

    NASA Astrophysics Data System (ADS)

    Ozevin, Didem; Li, Zhong; Heidary, Zahra

    2011-04-01

    Aluminum alloy 7075-T6 is a commonly used material in aircraft industry. A crack usually initiates at the edge of a fastener hole, and it can affect the maintenance schedule and reduce the life of an aircraft structure significantly. The fatigue property of the material has been researched widely to develop methods and models for predicting fatigue crack growth under random loading. From the point of damage tolerance design, the inspection technique of a crack for an aircraft structure is very important because it can be used to determine the inspection period of the aircraft structure. The acoustic emission (AE) technique is a nondestructive testing (NDT) method that is able to monitor damage initiation and progression in real time. Understanding the early stage of AE signature due to the damage progression using small scale laboratory samples requires non-traditional data analysis approaches. In this study, 1mm thick Al-7075-T6 plates were tested under monotonic and fatigue loading. The initiation of damage progression using AE data was identified based on improved linear location algorithm and the result was verified using elasto-plastic finite element model. The improved location algorithm integrates dispersive characteristics of flexural waves and threshold independent approach to pick up the wave arrival time. In this paper, AE results in comparison with FE model under monotonic and fatigue loading will be presented. The comparison of traditional and improved location approaches will be shown. The approach for implementing the laboratory scale results in the large scale field testing will be discussed.

  20. Experiments and simulation for 6061-T6 aluminum alloy resistance spot welded lap joints

    NASA Astrophysics Data System (ADS)

    Florea, Radu Stefanel

    This comprehensive study is the first to quantify the fatigue performance, failure loads, and microstructure of resistance spot welding (RSW) in 6061-T6 aluminum (Al) alloy according to welding parameters and process sensitivity. The extensive experimental, theoretical and simulated analyses will provide a framework to optimize the welding of lightweight structures for more fuel-efficient automotive and military applications. The research was executed in four primary components. The first section involved using electron back scatter diffraction (EBSD) scanning, tensile testing, laser beam profilometry (LBP) measurements, and optical microscopy(OM) images to experimentally investigate failure loads and deformation of the Al-alloy resistance spot welded joints. Three welding conditions, as well as nugget and microstructure characteristics, were quantified according to predefined process parameters. Quasi-static tensile tests were used to characterize the failure loads in specimens based upon these same process parameters. Profilometer results showed that increasing the applied welding current deepened the weld imprints. The EBSD scans revealed the strong dependency between the grain sizes and orientation function on the process parameters. For the second section, the fatigue behavior of the RSW'ed joints was experimentally investigated. The process optimization included consideration of the forces, currents, and times for both the main weld and post-heating. Load control cyclic tests were conducted on single weld lap-shear joint coupons to characterize the fatigue behavior in spot welded specimens. Results demonstrate that welding parameters do indeed significantly affect the microstructure and fatigue performance for these welds. The third section comprised residual strains of resistance spot welded joints measured in three different directions, denoted as in-plane longitudinal, in-plane transversal, and normal, and captured on the fusion zone, heat affected zone

  1. The Glucuronic Acid Utilization Gene Cluster from Bacillus stearothermophilus T-6

    PubMed Central

    Shulami, Smadar; Gat, Orit; Sonenshein, Abraham L.; Shoham, Yuval

    1999-01-01

    A λ-EMBL3 genomic library of Bacillus stearothermophilus T-6 was screened for hemicellulolytic activities, and five independent clones exhibiting β-xylosidase activity were isolated. The clones overlap each other and together represent a 23.5-kb chromosomal segment. The segment contains a cluster of xylan utilization genes, which are organized in at least three transcriptional units. These include the gene for the extracellular xylanase, xylanase T-6; part of an operon coding for an intracellular xylanase and a β-xylosidase; and a putative 15.5-kb-long transcriptional unit, consisting of 12 genes involved in the utilization of α-d-glucuronic acid (GlcUA). The first four genes in the potential GlcUA operon (orf1, -2, -3, and -4) code for a putative sugar transport system with characteristic components of the binding-protein-dependent transport systems. The most likely natural substrate for this transport system is aldotetraouronic acid [2-O-α-(4-O-methyl-α-d-glucuronosyl)-xylotriose] (MeGlcUAXyl3). The following two genes code for an intracellular α-glucuronidase (aguA) and a β-xylosidase (xynB). Five more genes (kdgK, kdgA, uxaC, uxuA, and uxuB) encode proteins that are homologous to enzymes involved in galacturonate and glucuronate catabolism. The gene cluster also includes a potential regulatory gene, uxuR, the product of which resembles repressors of the GntR family. The apparent transcriptional start point of the cluster was determined by primer extension analysis and is located 349 bp from the initial ATG codon. The potential operator site is a perfect 12-bp inverted repeat located downstream from the promoter between nucleotides +170 and +181. Gel retardation assays indicated that UxuR binds specifically to this sequence and that this binding is efficiently prevented in vitro by MeGlcUAXyl3, the most likely molecular inducer. PMID:10368143

  2. Mechanical properties of dissimilar metal joints composed of DP 980 steel and AA 7075-T6

    SciTech Connect

    Squires, Lile; Lim, Yong Chae; Miles, Michael; Feng, Zhili

    2015-03-18

    In this study, a solid state joining process, called friction bit joining, was used to spot weld aluminium alloy 7075-T6 to dual phase 980 steel. Lap shear failure loads for specimens without adhesive averaged ~10kN, while cross-tension specimens averaged 2·8 kN. Addition of adhesive with a thickness up to 500 μm provided a gain of ~50% to lap shear failure loads, while a much thinner layer of adhesive increased cross-tension failure loads by 20%. Microstructures of the welds were martensitic, but the hardness of the joining bit portion was greater than that of the DP 980, owing to its higher alloy content. Softening in the heat affected zone of a welded joint appeared to be relatively small, though it was enough to cause nugget pullout failures in some lap shear tension specimens. Finally, other failures in lap shear tension were interfacial, while all of the failures in cross-tension were interfacial.

  3. Erosion of aluminum 6061-T6 under cavitation attack in mineral oil and water

    NASA Technical Reports Server (NTRS)

    Rao, B. C. S.; Buckley, D. H.

    1985-01-01

    Studies of the erosion of aluminum 6061-T6 under cavitation attack in distilled water, ordinary tap water and a viscous mineral oil are presented. The mean depth of penetration for the mineral oil was about 40 percent of that for water at the end of a 40 min test. The mean depth of penetration and its rate did not differ significantly for distilled and tap water. The mean depth of penetration rate for both distilled and tap water increased to a maximum and then decreased with test duration, while that for mineral oil had a maximum during the initial period. The ratio h/2a of the pit depth h to the pit diameter 2a varied from 0.04 to 0.13 in water and from 0.06 to 0.20 in mineral oil. Scanning electron microscopy indicates that the pits are initially formed over the grain boundaries and precipitates while the surface grains are deformed under cavitation attack.

  4. Microstructure and Mechanical Behavior of Al 7075-T6 Subjected to Shallow Cryogenic Treatment

    NASA Astrophysics Data System (ADS)

    Mohan, K.; Suresh, J. A.; Ramu, Palaniappan; Jayaganthan, R.

    2016-06-01

    The effect of shallow cryogenic treatment (SCT) on the microstructure and mechanical properties of Al7075-T6 is investigated in the present work. The alloy was subjected to shallow CT at -80 °C for 72 h. Mechanical tests such as Vickers hardness test, tensile, and fatigue tests were performed on both native and treated samples. It was observed that the mechanical properties such as hardness, yield strength, and ultimate tensile strength increased by about 30, 17, and 7%, respectively, for the treated sample. The treated alloy was characterized by using the techniques such as optical microscopy, electron back scattered diffraction (EBSD), energy-dispersive x-ray spectroscopy (EDS), and transmission electron microscopy (TEM) to observe the changes in the microstructural features. EBSD results show precipitation, better distribution of second-phase particles, and higher dislocation density in the treated alloy as compared to the untreated alloy. The treatment imparts improved hardness and strength to the alloy due to precipitation hardening and high dislocation density. Fracture morphologies of the treated and the native samples were characterized by using scanning electron microscopy and it was observed that the striations were denser in the treated sample justifying the higher fatigue strength.

  5. Vertical Compensation Friction Stir Welding of 6061-T6 Aluminum Alloy

    NASA Astrophysics Data System (ADS)

    Ji, Shude; Meng, Xiangchen; Xing, Jingwei; Ma, Lin; Gao, Shuangsheng

    2016-09-01

    Vertical compensation friction stir welding (VCFSW) was proposed in order to solve the adverse effect caused by a big gap at the interface between two welded workpieces. VCFSW was successfully applied to weld 6061-T6 aluminum alloy with the thickness of 4 mm, while 2024-T4 aluminum alloy was selected as a rational compensation material. The results show that VCFSW is difficult to get a sound joint when the width of strip is no less than 1.5 mm. Decreasing the welding speed is beneficial to break compensation strip into pieces and then get higher quality joint. When the width of strip is 1 mm, the tensile strength and elongation of joint at the welding speed of 50 mm/min and rotational velocity of 1,800 rpm reach the maximum values of 203 MPa and 5.2%, respectively. Moreover, the addition of 2024-T4 alloy plays a strengthening effect on weld zone (WZ) of VCFSW joint. The fracture surface morphology of joint consisting of amounts of dimples exhibits ductile fracture.

  6. 2MASS J20261584-2943124: AN UNRESOLVED L0.5 + T6 SPECTRAL BINARY

    SciTech Connect

    Gelino, Christopher R.; Burgasser, Adam J.

    2010-07-15

    We identify the L dwarf 2MASS J20261584-2943124 as an unresolved spectral binary, based on low-resolution, near-infrared spectroscopy from IRTF/SpeX. The data reveal a peculiar absorption feature at 1.6 {mu}m, previously noted in the spectra of other very low-mass spectral binaries, which likely arises from overlapping FeH and CH{sub 4} absorption bands in the blended light of an L dwarf/T dwarf pair. Spectral template matching analysis indicates component types of L0.5 and T6, with relative brightness {Delta}H = 4.2 {+-} 0.6. Laser guide star adaptive optics imaging observations with Keck/NIRC2 fail to resolve the source, indicating a maximum separation at the observing epoch of 0.''25, or a projected separation of 9 AU assuming a distance of 36 {+-} 5 pc. With an age that is likely to be relatively older ({approx}>5 Gyr) based on the system's large V{sub tan} and mass ratio arguments, the relative motion of the potentially 'massive' (0.06-0.08 M{sub sun}) components of 2MASS J2026-2943 may be detectable through radial velocity variations, like its earlier-type counterpart 2MASS J03202839-0446358 (M8+T5), providing dynamical mass measurements that span the hydrogen burning limit.

  7. The effects of pitting on fatigue crack nucleation in 7075-T6 aluminum alloy

    NASA Technical Reports Server (NTRS)

    Ma, LI; Hoeppner, David W.

    1994-01-01

    A high-strength aluminum alloy, 7075-T6, was studied to quantitatively evaluate chemical pitting effects of its corrosion fatigue life. The study focused on pit nucleation, pit growth, and fatigue crack nucleation. Pitting corrosion fatigue experiments were conducted in 3.5 percent NaCl aqueous solution under constant amplitude sinusoidal loading at two frequencies, 5 and 20 Hz. Smooth and unnotched specimens were used in this investigation. A video recording system was developed to allow in situ observation of the surface changes of the specimens during testing. The results indicated that pitting corrosion considerably reduces the fatigue strength by accelerating fatigue crack nucleation. A metallographic examination was conducted on the specimens to evaluate the nature of corrosion pits. First, the actual shapes of the corrosion pits were evaluated by cross-sectioning the pits. Secondly, the relation between corrosion pits and microstructure was also investigated. Finally, the possibility of another corrosion mechanism that might be involved in pitting was explored in this investigation. The fractography of the tested specimens showed that corner corrosion pits were responsible for fatigue crack nucleation in the material due to the associated stress concentration. The pits exhibited variance of morphology. Fatigue life for the experimental conditions appeared to be strongly dependent on pitting kinetics and the crack nucleation stage.

  8. Mechanical properties of dissimilar metal joints composed of DP 980 steel and AA 7075-T6

    DOE PAGES

    Squires, Lile; Lim, Yong Chae; Miles, Michael; Feng, Zhili

    2015-03-18

    In this study, a solid state joining process, called friction bit joining, was used to spot weld aluminium alloy 7075-T6 to dual phase 980 steel. Lap shear failure loads for specimens without adhesive averaged ~10kN, while cross-tension specimens averaged 2·8 kN. Addition of adhesive with a thickness up to 500 μm provided a gain of ~50% to lap shear failure loads, while a much thinner layer of adhesive increased cross-tension failure loads by 20%. Microstructures of the welds were martensitic, but the hardness of the joining bit portion was greater than that of the DP 980, owing to its highermore » alloy content. Softening in the heat affected zone of a welded joint appeared to be relatively small, though it was enough to cause nugget pullout failures in some lap shear tension specimens. Finally, other failures in lap shear tension were interfacial, while all of the failures in cross-tension were interfacial.« less

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

    NASA Astrophysics Data System (ADS)

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

    2016-02-01

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

  10. Structural Conservation, Variability, and Immunogenicity of the T6 Backbone Pilin of Serotype M6 Streptococcus pyogenes

    PubMed Central

    Moreland, Nicole J.; Loh, Jacelyn M.; Bell, Anita; Atatoa Carr, Polly; Proft, Thomas; Baker, Edward N.

    2014-01-01

    Group A streptococcus (GAS; Streptococcus pyogenes) is a Gram-positive human pathogen that causes a broad range of diseases ranging from acute pharyngitis to the poststreptococcal sequelae of acute rheumatic fever. GAS pili are highly diverse, long protein polymers that extend from the cell surface. They have multiple roles in infection and are promising candidates for vaccine development. This study describes the structure of the T6 backbone pilin (BP; Lancefield T-antigen) from the important M6 serotype. The structure reveals a modular arrangement of three tandem immunoglobulin-like domains, two with internal isopeptide bonds. The T6 pilin lysine, essential for polymerization, is located in a novel VAKS motif that is structurally homologous to the canonical YPKN pilin lysine in other three- and four-domain Gram-positive pilins. The T6 structure also highlights a conserved pilin core whose surface is decorated with highly variable loops and extensions. Comparison to other Gram-positive BPs shows that many of the largest variable extensions are found in conserved locations. Studies with sera from patients diagnosed with GAS-associated acute rheumatic fever showed that each of the three T6 domains, and the largest of the variable extensions (V8), are targeted by IgG during infection in vivo. Although the GAS BP show large variations in size and sequence, the modular nature of the pilus proteins revealed by the T6 structure may aid the future design of a pilus-based vaccine. PMID:24778112

  11. Structural insights into methanol-stable variants of lipase T6 from Geobacillus stearothermophilus.

    PubMed

    Dror, Adi; Kanteev, Margarita; Kagan, Irit; Gihaz, Shalev; Shahar, Anat; Fishman, Ayelet

    2015-11-01

    Enzymatic production of biodiesel by transesterification of triglycerides and alcohol, catalyzed by lipases, offers an environmentally friendly and efficient alternative to the chemically catalyzed process while using low-grade feedstocks. Methanol is utilized frequently as the alcohol in the reaction due to its reactivity and low cost. However, one of the major drawbacks of the enzymatic system is the presence of high methanol concentrations which leads to methanol-induced unfolding and inactivation of the biocatalyst. Therefore, a methanol-stable lipase is of great interest for the biodiesel industry. In this study, protein engineering was applied to substitute charged surface residues with hydrophobic ones to enhance the stability in methanol of a lipase from Geobacillus stearothermophilus T6. We identified a methanol-stable variant, R374W, and combined it with a variant found previously, H86Y/A269T. The triple mutant, H86Y/A269T/R374W, had a half-life value at 70 % methanol of 324 min which reflects an 87-fold enhanced stability compared to the wild type together with elevated thermostability in buffer and in 50 % methanol. This variant also exhibited an improved biodiesel yield from waste chicken oil compared to commercial Lipolase 100L® and Novozyme® CALB. Crystal structures of the wild type and the methanol-stable variants provided insights regarding structure-stability correlations. The most prominent features were the extensive formation of new hydrogen bonds between surface residues directly or mediated by structural water molecules and the stabilization of Zn and Ca binding sites. Mutation sites were also characterized by lower B-factor values calculated from the X-ray structures indicating improved rigidity.

  12. Recovery of Mechanical Properties of a 6061-T6 Aluminum Weld by Heat Treatment After Welding

    NASA Astrophysics Data System (ADS)

    Pérez, Javier Serrano; Ambriz, Ricardo Rafael; López, Francisco Fernando Curiel; Vigueras, David Jaramillo

    2016-07-01

    The dilution effects in welds of a 6061-T6 (Al-Si-Mg) alloy obtained by the modified indirect electric arc (MIEA), using an ER4043 filler metal (Al-Si), and postweld heat treatment (PWHT) were analyzed. The soft zone (55 to 70 HV0.1) formed by the microstructural transformation in the heat-affected zone (HAZ) was eliminated. The hardness measurements were presented on a traditional microhardness profile and mapping representation. A hardening effect of the fusion zone was observed; the hardness values were above 120 HV0.1 and tended to be uniform. This behavior could be attributed to the chemical composition of the filler metal, the Mg migration from the base to the weld metal, and the reversible process of the PWHT, which promotes precipitation hardening. Improvement for yield (260 MPa) and tensile strength (310 MPa) of the MIEA joints was observed; these values were similar to those obtained for the base metal. However, the presence of porosity in the fusion zone limits the ductility of the joints (4.3 pct). Even though the yield and tensile strengths of the base metal and welded joints were similar, the stress concentration due to porosity in the weld metal generated data dispersion in fatigue life. As a consequence, the high-cycle fatigue life decreases with respect to the base metal. In contrast, when the crack propagates under elastic conditions, the crack-tip singularity is affected by the porosity in the weld metal (stress liberator). This aspect, in conjunction with the hardening effect in joints subjected to PWHT, improves the fatigue crack growth rate when compared to the as-welded condition.

  13. Identification of modes of fracture in a 2618-T6 aluminum alloy using stereophotogrammetry

    SciTech Connect

    Salas Zamarripa, A.; Mata, M.P. Guerrero; Morales, M. Castillo; Beber-Solano, T.P.

    2011-12-15

    The identification and the development of a quantification technique of the modes of fracture in fatigue fracture surfaces of a 2618-T6 aluminum alloy were developed during this research. Fatigue tests at room and high temperature (230 Degree-Sign C) were carried out to be able to compare the microscopic fractographic features developed by this material under these testing conditions. The overall observations by scanning electron microscopy (SEM) of the fracture surfaces showed a mixture of transgranular and ductile intergranular fracture. The ductile intergranular fracture contribution appears to be more significant at room temperature than at 230 Degree-Sign C. A quantitative methodology was developed to identify and to measure the contribution of these microscopic fractographic features. The technique consisted of a combination of stereophotogrammetry and image analysis. Stereo-pairs were randomly taken along the crack paths and were then analyzed using the profile module of MeX software. The analysis involved the 3-D surface reconstruction, the trace of primary profile lines in both vertical and horizontal directions within the stereo-pair area, the measurements of the contribution of the modes of fracture in each profile, and finally, the calculation of the average contribution in each stereo-pair. The technique results confirmed a higher contribution of ductile intergranular fracture at room temperature than at 230 Degree-Sign C. Moreover, there was no indication of a direct relationship between this contribution and the strain amplitudes range applied during the fatigue testing. - Highlights: Black-Right-Pointing-Pointer Stereophotogrammetry and image analysis as a measuring tool of modes of fracture in fatigue fracture surfaces. Black-Right-Pointing-Pointer A mixture of ductile intergranular and transgranular fracture was identified at room temperature and 230 Degree-Sign C testing. Black-Right-Pointing-Pointer Development of a quantitative methodology to

  14. Structural insights into methanol-stable variants of lipase T6 from Geobacillus stearothermophilus.

    PubMed

    Dror, Adi; Kanteev, Margarita; Kagan, Irit; Gihaz, Shalev; Shahar, Anat; Fishman, Ayelet

    2015-11-01

    Enzymatic production of biodiesel by transesterification of triglycerides and alcohol, catalyzed by lipases, offers an environmentally friendly and efficient alternative to the chemically catalyzed process while using low-grade feedstocks. Methanol is utilized frequently as the alcohol in the reaction due to its reactivity and low cost. However, one of the major drawbacks of the enzymatic system is the presence of high methanol concentrations which leads to methanol-induced unfolding and inactivation of the biocatalyst. Therefore, a methanol-stable lipase is of great interest for the biodiesel industry. In this study, protein engineering was applied to substitute charged surface residues with hydrophobic ones to enhance the stability in methanol of a lipase from Geobacillus stearothermophilus T6. We identified a methanol-stable variant, R374W, and combined it with a variant found previously, H86Y/A269T. The triple mutant, H86Y/A269T/R374W, had a half-life value at 70 % methanol of 324 min which reflects an 87-fold enhanced stability compared to the wild type together with elevated thermostability in buffer and in 50 % methanol. This variant also exhibited an improved biodiesel yield from waste chicken oil compared to commercial Lipolase 100L® and Novozyme® CALB. Crystal structures of the wild type and the methanol-stable variants provided insights regarding structure-stability correlations. The most prominent features were the extensive formation of new hydrogen bonds between surface residues directly or mediated by structural water molecules and the stabilization of Zn and Ca binding sites. Mutation sites were also characterized by lower B-factor values calculated from the X-ray structures indicating improved rigidity. PMID:26026940

  15. Ratcheting fatigue behaviour of Al-7075 T6 alloy: Influence of stress parameters

    NASA Astrophysics Data System (ADS)

    Amarnath, Lala; Bhattacharjee, Antara; Dutta, K.

    2016-02-01

    The use of aluminium and aluminium based alloys are increasing rapidly on account of its high formability, good thermal and electrical conductivity, high strength and lightness. Aluminium alloys are extensively used in aerospace, automobile, marine and space research industries and are also put into structural applications where chances of fatigue damage cannot be ruled out. In the current work, it is intended to study the ratcheting fatigue behavior of 7075-T6 aluminium alloy at room temperature. This Al alloy is potentially used in aviation, marine and automotive components as well as in bicycle parts, rock mounting equipment and parts of ammunition where there is every chance of failure of the parts due to deformation caused by ratcheting. Ratcheting is the process of accruement of plastic stain produced when a component is subjected to asymmetric cyclic loading under the influence of low cycle fatigue. To accomplish the requirements of the projected research, stress-controlled cyclic loading experiments were done using a ±250 kN servo-hydraulic universal testing machine (Instron: 8800R). The effect of stress parameters such as mean stress and stress amplitude were investigated on the ratcheting behavior of the selected aluminium alloy. It was observed that, ratcheting strain increased with increase in the value of stress amplitude at any constant mean stress while a saturation in strain accumulation attained in the investigated material after around 10-20 cycles, under all test conditions. The analyses of hysteresis loop generated during cyclic loading indicate that the material exhibits cyclic hardening in the initial fifty cycles which gets softened in further loading up to about 70-80 cycles and finally attains a steady state. The increase in the ratcheting strain value with stress parameters happens owing to increased deformation domain during cycling. The cyclic hardening accompanied by softening is correlated with characteristic precipitation features of

  16. An Overview of the T6 Gridded Ion Propulsion System Pre-Development Activities for Alpha-Bus

    NASA Astrophysics Data System (ADS)

    Huddleson, J.; Brandon-Cox, J.; Wallace, N.; Palencia, J.

    2004-10-01

    This paper provides an overview of the Gridded Ion Propulsion System pre-development activities, partially funded by ESA, which QinetiQ and its partners are performing for the European AlphaBus programme. The system is based on the QinetiQ T6 Kaufman Ion Thruster, and includes the power processing units and flow control units that power and provide propellant to the thrusters. An overview of the T6 thruster technology and heritage is provided. This is followed by an overview of how the system hardware is being developed for this application, including how QinetiQ will maximise the benefit of common heritage with parallel T6 programmes, such as the Bepi-Colombo Technology Demonstration Activity.

  17. Diversity of the biosynthesis pathway for threonylcarbamoyladenosine (t6A), a universal modification of tRNA

    PubMed Central

    Thiaville, Patrick C; Iwata-Reuyl, Dirk; de Crécy-Lagard, Valérie

    2014-01-01

    The tRNA modification field has a rich literature covering biochemical analysis going back more than 40 years, but many of the corresponding genes were only identified in the last decade. In recent years, comparative genomic-driven analysis has allowed for the identification of the genes and subsequent characterization of the enzymes responsible for N6-threonylcarbamoyladenosine (t6A). This universal modification, located in the anticodon stem-loop at position 37 adjacent to the anticodon of tRNAs, is found in nearly all tRNAs that decode ANN codons. The t6A biosynthesis enzymes and synthesis pathways have now been identified, revealing both a core set of enzymes and kingdom-specific variations. This review focuses on the elucidation of the pathway, diversity of the synthesis genes, and proposes a new nomenclature for t6A synthesis enzymes. PMID:25629598

  18. Investigating the presence of predatory bacteria on algal bloom samples using a T6SS gene marker.

    NASA Astrophysics Data System (ADS)

    Hendricks, J.; Sison-Mangus, M.; Mehic, S.; McMahon, E.

    2015-12-01

    Predation is considered to be a major driving force in evolution and ecology, which has been observed affecting individual organisms, communities, and entire ecosystems. The type VI secretion system (T6SS) is an intermembranal protein complex identified in certain bacteria, which appears to have evolved strictly as a mechanism of predation. The effects of bacteria on phytoplankton physiology are still understudied, however, studies have shown that the interactions between bacteria that inhabit the phycosphere of phytoplankton can possibly result in coevolution of native host and microbiota. It is unclear if bacteria can prey upon other bacteria to gain advantages during periods of high phytoplankton density. Here, we investigate the predatory interactions between bacteria and analyze environmental samples for the presence of predatory bacterial genes in an effort to understand bacteria-bacteria and phytoplankton interactions during algal blooms. DNA were extracted from bacterial samples collected weekly from size-fractionated samples using 3.0 um and 0.2 um membrane filters at the Santa Cruz wharf. PCR amplification and gel visualization for the presence of T6SS gene was carried out on bloom and non-bloom samples. Moreover, we carried out a lab- based experiment to observe bacteria-bacteria interaction that may hint for the presence of predatory behavior between bacterial taxa. We observed what appeared to be a predatory biofilm formation between certain bacterial species. These bacteria, however, did not contain the T6SS genes. On the contrary the T6SS gene was discovered in some of the bloom samples gathered from the Santa Cruz wharf. It is still unclear if the predatory mechanisms facilitate the abundance of certain groups of bacteria that contain the T6SS genes during algal blooms, but our evidence suggest that bacterial predation through T6SS mechanism is present during bloom events.

  19. Regulation of nucleotide and pentose synthesis in resting and stimulated 3T6 fibroblasts.

    PubMed

    Buchanan, J M; Smith, M L; Smith, R J

    1982-01-01

    A two-step procedure has been used to follow the activation of one metabolic system involved in the return of cells to a proliferative state after resting in a Go state as a result of serum limitation. One feature of the resting state is a limited capacity to synthesize nucleotides. The limitation apparently is in the rate of synthesis of 5-phosphoribosylpyrophosphate from glucose and indirectly in the capacity of the resting cells to turn over the triphosphopyridine nucleotide pair, NADPH:NADP+. A reaction utilizing NADPH is apparently greatly diminished in resting cells and is substantially increased by only brief contact of cells with the hormonal elements in dialyzed calf serum. Insulin together with platelet-derived growth factor can substitute for calf serum. Aside from stimulating the turnover of the pyridine nucleotide coenzyme pair, serum also stimulates the utilization and reformation of ATP, principally from AMP. Among the NADPH-linked reactions that have been examined for their physiological significance in the initiation of growth stimulation are two steps in the conversion of glutamate to proline in the cytoplasm. Pyrroline 5-carboxylate, an intermediate in this metabolic pathway, has been shown to stimulate PRPP synthesis when added to cultures of resting 3T6 cells. Proline, the product of the reduction of this 5-membered heterocycle is also a stimulant of PRPP synthesis. In addition, dehydroascorbic acid is a potent stimulant of PRPP synthesis. As a working hypothesis, we are exploring the role of a series of reactions that form a pyrroline 5-carboxylate/proline cycle operating between the cytoplasm and mitochondria. The net result is the oxidation of NADPH by molecular oxygen to yield NADP+ and water. The NADP+ is then used in the hexose monophosphate pathway for the conversion of glucose to PRPP. We wish to determine whether dehydroascorbate is operating in this cycle as an oxidant of proline in the mitochondria or whether it participates in some

  20. Renal cell carcinomas with t(6;11) (p21;q12): presentation of two cases with computed tomography findings.

    PubMed

    Zhao, YiJun; Yao, Jin; Chen, Ni; Zeng, Hao; Zhang, Wei

    2015-06-01

    Renal cell carcinomas with t(6;11) (p21;q12) translocation are extremely rare and primarily affect children and young adults. To our knowledge, there has been no case report focusing on the imaging manifestations in the existing literature. Hence, we describe the computed tomography findings of two young adults.

  1. Estudio numerico y experimental del proceso de soldeo MIG sobre la aleacion 6063--T5 utilizando el metodo de Taguchi

    NASA Astrophysics Data System (ADS)

    Meseguer Valdenebro, Jose Luis

    improvement on mechanical properties in aluminum metal joint. Los procesos de soldadura por arco electrico representan unas de las tecnicas mas utilizadas en los procesos de fabricacion de componentes mecanicos en la industria moderna. Los procesos de soldeo por arco se han adaptado a las necesidades actuales, haciendose un modo de fabricacion flexible y versatil. Los resultados obtenidos numericamente en el proceso de soldadura son validados experimentalmente. Los principales metodos numericos mas empleados en la actualidad son tres, metodo por diferencias finitas, metodos por elementos finitos y metodo por volumenes finitos. El metodo numerico mas empleado para el modelado de uniones soldadas, es el metodo por elementos finitos, debido a que presenta una buena adaptacion a las condiciones geometricas y de contorno ademas de que existe una diversidad de programas comerciales que utilizan el metodo por elementos finitos como base de calculo. Este trabajo de investigacion presenta un estudio experimental de una union soldada mediante el proceso MIG de la aleacion de aluminio 6063-T5. El metodo numerico se valida experimentalmente aplicando el metodo de los elementos finitos con el programa de calculo ANSYS. Los resultados experimentales obtenidos son: las curvas de enfriamiento, el tiempo critico de enfriamiento t4/3, geometria del cordon, microdurezas obtenidas en la union soldada, zona afectada termicamente y metal base, dilucion del proceso, areas criticas intersecadas entre las curvas de enfriamiento y la curva TTP. Los resultados numericos son: las curvas del ciclo termico, que representan tanto el calentamiento hasta alcanzar la temperatura maxima y un posterior enfriamiento. Se calculan el tiempo critico de enfriamiento t4/3, el rendimiento termico y se representa la geometria del cordon obtenida experimentalmente. La zona afectada termicamente se obtiene diferenciando las zonas que se encuentran a diferentes temperaturas, las areas criticas intersecadas entre las

  2. Laser peening without coating on aluminum alloy Al-6061-T6 using low energy Nd:YAG laser

    NASA Astrophysics Data System (ADS)

    Sathyajith, S.; Kalainathan, S.; Swaroop, S.

    2013-02-01

    The present study investigates the effect of laser peening without coating on aluminum alloy Al-6061-T6 with a 300 mJ infrared laser. The surface topography, microstructure, surface topography, surface residual stress and micro-hardness of peened and unpeened surfaces were studied. The study shows that laser peening without coating can significantly improve surface compressive stress and micro-hardness with trivial increase in surface roughness. Microstructure evaluation confirmed there was no near surface solidification after LPwC.

  3. UV spectroscopic identification and thermodynamic analysis of protonated third strand deoxycytidine residues at neutrality in the triplex d(C(+)-T)6:[d(A-G)6.d(C-T)6]; evidence for a proton switch.

    PubMed Central

    Lavelle, L; Fresco, J R

    1995-01-01

    Near-UV difference spectral analysis of the triplex formed from d(C-T)6 and d(A-G)6.d(C-T)6 in neutral and acidic solution shows that the third strand dC residues are protonated at pH 7.0, far above their intrinsic pKa. Additional support for ion-dipole interactions between the third strand dC residues and the G.C target base pairs comes from reduced positive dependence of triplet stability on ionic strength below 0.9 M Na+, inverse dependence above 0.9 M Na+ and strong positive dependence on hydrogen ion concentration. Molecular modeling (AMBER) of C:G.C and C+:G.C base triplets with the third strand base bound in the Hoogsteen geometry shows that only the C+:G.C triplet is energetically feasible. van't Hoff analysis of the melting of the triplex and target duplex shows that between pH 5.0 and 8.5 in 0.15 M NaCl/0.005 M MgCl2 the enthalpy of melting (delta H degree obs) varies from 5.7 to 6.6 kcal.mol-1 for the duplex in a duplex mixture and from 7.3 to 9.7 kcal.mol-1 for third strand dissociation in the triplex mixture. We have extended the condensation-screening theory of Manning to pH-dependent third strand binding. In this development we explicitly include the H+ contribution to the electrostatic free energy and obtain [formula: see text]. The number of protons released in the dissociation of the third strand from the target duplex at pH 7.0, delta n2, is thereby calculated to be 5.5, in good agreement with approximately six third strand dc residues per mole of triplex. This work shows that when third strand binding requires protonated residues that would otherwise be neutral, triplex formation and dissociation are mediated by proton uptake and release, i.e., a proton switch. As a by-product of this study, we have found that at low pH the Watson-Crick duplex d(A-G)6.d(C-T)6 undergoes a transition to a parallel Hoogsteen duplex d(A-G)6.d(C(+)-T)6. PMID:7651830

  4. A Williams syndrome patient with a familial t(6;7) translocation and deletion of the elastin gene

    SciTech Connect

    Pober, B.R.; Gibson, L.H.; Yang-Feng, T.L.

    1994-09-01

    Discovery of a {open_quotes}balanced{close_quotes} reciprocal translocation [46,XX,t(6;7)(q11.2;q11.23)] on routine amniocentesis prompted clinical and cytogenetic study of additional family members. The same t(6;7) translocation was found in the clincally normal father and in a sibling with Williams syndrome (WS). WS had been diagnosed previously according to clinical criteria including distinctive facial features, supravalvar aortic stenosis requiring surgical repair, dental abnormalties and developmental delay. A clinically normal female was delivered and the translocation was confirmed with a cord blood specimen. Hemizygosity for the gene, elastin, (which has been mapped to the chromosome 7 translocation breakpoint, 7q11.23, in this family) appears to be a cause of WS. We therefore investigated whether the t(6;7) in the phenotypically normal father represented more than a simple reciprocal translocation. FISH using a chromosome 7 specific library revealed no differences between the cytogenetically identical, yet phenotypically distinct, father and son. Hybridization with a cosmid MR127D4 containing elastin sequence showed that the WS patient was missing one allele from the derivative chromosome 7 whereas both his mother and father had two copies of the elastin gene. This family indicates that the de novo loss of one copy of the elastin gene produces the recognizable phenotype of Williams syndrome. Molecular characterization (with additional probes) of the extent of this de novo deletion near the translocation breakpoint is in progress. This information will be valuable for defining the WS-critical region and will lead to a better understanding of the molecular basis for WS.

  5. Electrochemical investigation on the hydrogen permeation behavior of 7075-T6 Al alloy and its influence on stress corrosion cracking

    NASA Astrophysics Data System (ADS)

    Zheng, Chuan-bo; Yan, Bing-hao; Zhang, Ke; Yi, Guo

    2015-07-01

    The hydrogen permeation behavior and stress corrosion cracking (SCC) susceptibility of precharged 7075-T6 Al alloy were investigated in this paper. Devanthan-Stachurski (D-S) cell tests were used to measure the apparent hydrogen diffusivity and hydrogen permeation current density of specimens immersed in 3.5wt% NaCl solution. Electrochemical experiment results show that the SCC susceptibility is low during anodic polarization. Both corrosion pits and hydrogen-induced cracking are evident in scanning electron microscope images after the specimens have been charging for 24 h.

  6. An extensive allelic series of Drosophila kae1 mutants reveals diverse and tissue-specific requirements for t6A biogenesis.

    PubMed

    Lin, Ching-Jung; Smibert, Peter; Zhao, Xiaoyu; Hu, Jennifer F; Ramroop, Johnny; Kellner, Stefanie M; Benton, Matthew A; Govind, Shubha; Dedon, Peter C; Sternglanz, Rolf; Lai, Eric C

    2015-12-01

    N(6)-threonylcarbamoyl-adenosine (t6A) is one of the few RNA modifications that is universally present in life. This modification occurs at high frequency at position 37 of most tRNAs that decode ANN codons, and stabilizes cognate anticodon-codon interactions. Nearly all genetic studies of the t6A pathway have focused on single-celled organisms. In this study, we report the isolation of an extensive allelic series in the Drosophila ortholog of the core t6A biosynthesis factor Kae1. kae1 hemizygous larvae exhibit decreases in t6A that correlate with allele strength; however, we still detect substantial t6A-modified tRNAs even during the extended larval phase of null alleles. Nevertheless, complementation of Drosophila Kae1 and other t6A factors in corresponding yeast null mutants demonstrates that these metazoan genes execute t6A synthesis. Turning to the biological consequences of t6A loss, we characterize prominent kae1 melanotic masses and show that they are associated with lymph gland overgrowth and ectopic generation of lamellocytes. On the other hand, kae1 mutants exhibit other phenotypes that reflect insufficient tissue growth. Interestingly, whole-tissue and clonal analyses show that strongly mitotic tissues such as imaginal discs are exquisitely sensitive to loss of kae1, whereas nonproliferating tissues are less affected. Indeed, despite overt requirements of t6A for growth of many tissues, certain strong kae1 alleles achieve and sustain enlarged body size during their extended larval phase. Our studies highlight tissue-specific requirements of the t6A pathway in a metazoan context and provide insights into the diverse biological roles of this fundamental RNA modification during animal development and disease.

  7. Simulation of d(7Be,t)6Be with Fresco, and investigation of resonant states of 6Be

    NASA Astrophysics Data System (ADS)

    Pelham, Timothy

    2008-10-01

    An ongoing study is presented into the resonant states of 6Be via simulation of d(7Be,t)6Be with Fresco to investigate the results of ``Searching for resonances in the unbound 6Be nucleus'' a paper by K.Y. Chae [1]. In this paper the d(7Be,t)6Be reaction was studied to search for resonances in the 6Be nucleus that may be used to increase our knowledge of the 3He(3He,2p)4He reaction. A 100-MeV 7Be beam from the Holifield Radioactive Ion Beam Facility (HRIBF) was used to bombard CD2 targets, and tritons were detected by the Silicon Detector Array (SIDAR). It was concluded that a combination of reaction mechanisms are necessary to account for the observed triton energy spectrum. This will be further investigated by simulating the various reaction mechanisms with Fresco to try to reproduce and explain these results. Preliminary Results will be presented.

  8. An evaluation of the T-6A Texan (JPATS) initial functional performance of the CFIS laser assemblies

    NASA Astrophysics Data System (ADS)

    Blachowski, Thomas J.; Eccard, George; Thom, Travis

    2009-08-01

    The Indian Head Division, Naval Surface Warfare Center (IHDIV, NSWC) CAD Engineering Division is conducting a program to evaluate the laser components which comprise the Canopy Fracturing Initiation System (CFIS) currently installed on the T-6A Texan or JPATS (Joint Primary Aircraft Training System) aircraft. The T-6A Texan is the first aircraft used by the military to train future pilots. The CFIS is an element of the pilot emergency escape system which weakens the canopy in the path of the ejection seat. The CFIS is comprised of three differing configurations (Internal, External, and Seat Motion) which generate a laser pulse that is distributed through a fiber optic energy transmission system. This pulse, in turn, initiates explosive components which weaken the respective canopies. All of the CFIS laser types are flashlamp-pumped, neodymium glass lasers which are located at various positions in the aircraft cockpit area. This paper builds on the previous 2008 SPIE paper (Conference 7070) and presents further CAD Engineering Division test results and analysis which were utilized to evaluate the functional performance of the three CFIS laser signal generators after their being installed fleet applications over a period of time.

  9. A study of fatigue and fracture in 7075-T6 aluminum alloy in vacuum and air environments

    NASA Technical Reports Server (NTRS)

    Hudson, C. M.

    1973-01-01

    Axial load fatigue life, fatigue-crack propagation, and fracture toughness experiments were conducted on sheet specimens made of 7075-T6 aluminum alloy. These experiments were conducted at pressures ranging from atmospheric to 5 x 10 to the minus 8th torr. Analysis of the results from the fatigue life experiments indicated that for a given stress level, lower air pressures produced longer fatigue lives. At a pressure of 5 x 10 to the minus 8th torr fatigue lives were 15 or more times as long as at atmospheric pressure. Analysis of the results from the fatigue crack propagation experiments indicated that for small stress intensity factor ranges the fatigue crack propagation rates were up to twice as high at atmospheric pressure as in vacuum. The fracture toughness of 7075-T6 was unaffected by the vacuum environment. Fractographic examination showed that specimens tested in both vacuum and air developed fatigue striations. Considerably more striations developed on specimens tested at atmospheric pressure, however.

  10. Review of renal carcinoma with t(6;11)(p21;q12) with focus on clinical and pathobiological aspects.

    PubMed

    Kuroda, Naoto; Tanaka, Azusa; Sasaki, Naomi; Ishihara, Akira; Matsuura, Keiko; Moriyama, Masatsugu; Nagashima, Yoji; Inoue, Keiji; Petersson, Fredrik; Martignoni, Guido; Michal, Michal; Hes, Ondrej

    2013-06-01

    Recently, a new category of MiTF/TFE family translocation carcinomas of the kidney has been proposed. This category includes Xp11.2 renal cell carcinoma (RCC) and the t(6;11) RCC. These tumors share clinical, morphological, immunohistochemical and molecular genetic features. In this article, we review t(6;11) RCC. This tumor predominantly affects children and young adults. Macroscopically, the tumor generally forms a well circumscribed mass. Satellite nodules may be observed. Histologically, the tumor comprises large cells and small cells surrounded by basement membrane material. Immunohistochemically, tumor cells show nuclear immunolabeling for TFEB and usually express Cathepsin-K in the cytoplasm. Karyotyping detects the rearrangement between chromosome 6p21 and chromosome 11q12. Alpha-TFEB fusion can be detected by reverse transcriptase polymerase chain reaction (RT-PCR) or fluorescence in situ hybridization (FISH). Most cases affecting children and young adults seem to be indolent, but some adult cases have presented with metastasis or caused death. As previously reported cases remain limited to date, further examination in a large scale study will be needed in order to elucidate clinical behavior and molecular characteristics.

  11. Immunodeficiency in RFM/(T6xRFM)F1 mouse chimaeras with lethal host-versus-graft syndrome.

    PubMed Central

    Hard, R C; Montour, J L; Fuchs, B A

    1995-01-01

    Rather than central tolerance, the perinatal inoculation of related F1 hybrid spleen cells into inbred mice may result in host-versus-graft (HVG) reactions manifested as transient autoimmunity, or as a lethal immunodeficiency syndrome. RFM/(T6xRFM)F1 chimaeras with lethal disease die in 30 days with lymphosplenomegaly, immune complexes and impaired immune responses. The present studies used in vitro proliferation assays to show that the HVG reaction caused hyperplasia sufficient to account for the lymphosplenomegaly, while also causing severe impairment of splenic and nodal cell responses to concanavalin A (Con A) and to bacterial lipopolysaccharide (LPS). By 25 days, HVG mice could not distinguish between self and non-self as judged by mixed lymphocyte reactions (MLR) to RFM, (T6xRFM)F1 and third party A/J cells. There were no indications that host cells reactive to F1 donor cells had undergone clonal deletion, anergy or expansion. Flow cytometry revealed that donor T lymphocytes achieved stable engraftment, mostly in the nodes, despite the HVG reaction. Taken together with previous observations, these studies showed that HVG reactions in young parent F1/chimaeras can result in an immunodeficiency state which is characterized by an early appearing, profound and persistent impairment of both host and donor T and B cell functions. The results suggest that HVG reactions can contribute directly to immune deficits seen after clinical allogeneic bone marrow transplantation. PMID:7743676

  12. A molecular dynamics investigation into the size-dependent buckling behavior of a novel three-dimensional metallic carbon nanostructure (T6)

    NASA Astrophysics Data System (ADS)

    Ansari, R.; Ajori, S.; Hassani, R.

    2016-09-01

    The buckling behavior of a novel three-dimensional metallic carbon nanostructure known as T6 is investigated herein employing the molecular dynamics (MD) simulations. The models are prepared on the basis of two beam- and plate-like structures to study the effects of size and geometry on the critical buckling force and critical strain. It is observed that the range of critical force for the beam-like and plate-like T6 with different geometrical parameters is approximately identical. Moreover, it is demonstrated that the critical buckling force decreases and increases by increasing the length and the width of T6, respectively. Moreover, it is shown that critical strain of beam-like T6 decreases by increasing the length, whereas, in the case of plate-like T6, the critical strain only fluctuates around 2% by increasing the width. It is further found that the buckling parameters of T6 are not comparable with those of single-walled carbon nanotubes (SWCNTs) and graphene with a relatively similar dimension. The critical buckling force and critical strain of T6 are considerably smaller than those of SWCNT and larger than those of graphene.

  13. Application of Plant-Growth-Promoting Fungi Trichoderma longibrachiatum T6 Enhances Tolerance of Wheat to Salt Stress through Improvement of Antioxidative Defense System and Gene Expression

    PubMed Central

    Zhang, Shuwu; Gan, Yantai; Xu, Bingliang

    2016-01-01

    Soil salinity is a serious problem worldwide that reduces agricultural productivity. Trichoderma longibrachiatum T6 (T6) has been shown to promote wheat growth and induce plant resistance to parasitic nematodes, but whether the plant-growth-promoting fungi T6 can enhance plant tolerance to salt stress is unknown. Here, we determined the effect of plant-growth-promoting fungi T6 on wheat seedlings’ growth and development under salt stress, and investigated the role of T6 in inducing the resistance to NaCl stress at physiological, biochemical, and molecular levels. Wheat seedlings were inoculated with the strain of T6 and then compared with non-inoculated controls. Shoot height, root length, and shoot and root weights were measured on 15 days old wheat seedlings grown either under 150 mM NaCl or in a controlled setting without any NaCl. A number of colonies were re-isolated from the roots of wheat seedlings under salt stress. The relative water content in the leaves and roots, chlorophyll content, and root activity were significantly increased, and the accumulation of proline content in leaves was markedly accelerated with the plant growth parameters, but the content of leaf malondialdehyde under saline condition was significantly decreased. The antioxidant enzymes-superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in wheat seedlings were increased by 29, 39, and 19%, respectively, with the application of the strain of T6 under salt stress; the relative expression of SOD, POD, and CAT genes in these wheat seedlings were significantly up-regulated. Our results indicated that the strain of T6 ameliorated the adverse effects significantly, protecting the seedlings from salt stress during their growth period. The possible mechanisms by which T6 suppresses the negative effect of NaCl stress on wheat seedling growth may be due to the improvement of the antioxidative defense system and gene expression in the stressed wheat plants.

  14. Application of Plant-Growth-Promoting Fungi Trichoderma longibrachiatum T6 Enhances Tolerance of Wheat to Salt Stress through Improvement of Antioxidative Defense System and Gene Expression

    PubMed Central

    Zhang, Shuwu; Gan, Yantai; Xu, Bingliang

    2016-01-01

    Soil salinity is a serious problem worldwide that reduces agricultural productivity. Trichoderma longibrachiatum T6 (T6) has been shown to promote wheat growth and induce plant resistance to parasitic nematodes, but whether the plant-growth-promoting fungi T6 can enhance plant tolerance to salt stress is unknown. Here, we determined the effect of plant-growth-promoting fungi T6 on wheat seedlings’ growth and development under salt stress, and investigated the role of T6 in inducing the resistance to NaCl stress at physiological, biochemical, and molecular levels. Wheat seedlings were inoculated with the strain of T6 and then compared with non-inoculated controls. Shoot height, root length, and shoot and root weights were measured on 15 days old wheat seedlings grown either under 150 mM NaCl or in a controlled setting without any NaCl. A number of colonies were re-isolated from the roots of wheat seedlings under salt stress. The relative water content in the leaves and roots, chlorophyll content, and root activity were significantly increased, and the accumulation of proline content in leaves was markedly accelerated with the plant growth parameters, but the content of leaf malondialdehyde under saline condition was significantly decreased. The antioxidant enzymes-superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in wheat seedlings were increased by 29, 39, and 19%, respectively, with the application of the strain of T6 under salt stress; the relative expression of SOD, POD, and CAT genes in these wheat seedlings were significantly up-regulated. Our results indicated that the strain of T6 ameliorated the adverse effects significantly, protecting the seedlings from salt stress during their growth period. The possible mechanisms by which T6 suppresses the negative effect of NaCl stress on wheat seedling growth may be due to the improvement of the antioxidative defense system and gene expression in the stressed wheat plants. PMID:27695475

  15. Accidental finding of Hashimoto-like thyroiditis in male B.U.T. 6 turkeys at slaughter.

    PubMed

    Plesch, P; Schade, B; Breithaupt, A; Bellof, G; Kienzle, E

    2014-10-01

    In the context of a study on the tolerance of rapeseed meal in B.U.T. 6 turkeys, thyroid glands were histologically and immunohistochemically examined because of potential thyreostatic effects. In all groups including the controls with no rapeseed meal in their food, there was a high incidence of lymphocytic infiltration and thyroiditis (14% of thyroids with moderate to severe lymphocytic thyroiditis). Thirty per cent of mononuclear inflammatory cells were immunohistochemically identified as T cells. There were occasional accumulations of PAX-5 labelled cells, indicating germinal centre development. These lesions resemble Hashimoto's disease in humans. The effect on thyroid function is unknown. Mild hypothyreosis might enhance productivity but also explain dispositions towards diseases seen in context with thyroid dysfunction such as skin diseases (foot pad disease?) and cardiovascular problems. Further studies on thyroid function in these turkeys are needed.

  16. Fatigue and fracture behavior of an aluminum-lithium alloy 8090-T6 at ambient and cryogenic temperature

    NASA Astrophysics Data System (ADS)

    Xu, Y. B.; Wang, L.; Zhang, Y.; Wang, Z. G.; Hu, Q. Z.

    1991-03-01

    An investigation has been made of the fatigue and fracture behavior of an Al-Li-Cu-Mg-Zr 8090-T6 alloy at room (300 K) and liquid nitrogen (77 K) temperatures. The fatigue and fracture strengths, as well as ductility of the alloy, have been found to increase with decreasing temperature. The observations by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) reveal that the changes in fatigue and fracture behavior with temperature are considered to be associated with the change in the deformation and fracture modes. It has been found that the occurrence of the localized shear deformation bands in which the hard precipitates are sheared by moving dislocations is responsible for the reduction of fatigue and fracture strengths as well as ductility of the alloy at room temperature. However, the improvement of both strength and ductility of the alloy at liquid nitrogen temperature might be attributed to the deeper and larger delamination that occurred on the fracture surface.

  17. Corrosion behavior of the friction-stir-welded joints of 2A14-T6 aluminum alloy

    NASA Astrophysics Data System (ADS)

    Qin, Hai-long; Zhang, Hua; Sun, Da-tong; Zhuang, Qian-yu

    2015-06-01

    The corrosion behavior of friction-stir-welded 2A14-T6 aluminum alloy was investigated by immersion testing in immersion exfoliation corrosion (EXCO) solution. Electrochemical measurements (open circuit potential, potentiodynamic polarization curves, and electrochemical impedance spectroscopy), scanning electron microscopy, and energy dispersive spectroscopy were employed for analyzing the corrosion mechanism. The results show that, compared to the base material, the corrosion resistance of the friction-stir welds is greatly improved, and the weld nugget has the highest corrosion resistance. The pitting susceptibility originates from the edge of Al-Cu-Fe-Mn-Si phase particles as the cathode compared to the matrix due to their high self-corrosion potential. No corrosion activity is observed around the θ phase (Al2Cu) after 2 h of immersion in EXCO solution.

  18. Effect of Welding Speeds on Mechanical Properties of Level Compensation Friction Stir Welded 6061-T6 Aluminum Alloy

    NASA Astrophysics Data System (ADS)

    Wen, Quan; Yue, Yumei; Ji, Shude; Li, Zhengwei; Gao, Shuangsheng

    2016-04-01

    In order to eliminate the flash, arc corrugation and concave in weld zone, level compensation friction stir welding (LCFSW) was put forward and successfully applied to weld 6061-T6 aluminum alloy with varied welding speed at a constant tool rotational speed of 1,800 rpm in the present study. The glossy joint with equal thickness of base material can be attained, and the shoulder affected zone (SAZ) was obviously reduced. The results of transverse tensile test indicate that the tensile strength and elongation reach the maximum values of 248 MPa and 7.1% when the welding speed is 600 mm/min. The microhardness of weld nugget (WN) is lower than that of base material. The tensile fracture position locates at the heat affected zone (HAZ) of the advancing side (AS), where the microhardness is the minimum. The fracture surface morphology represents the typical ductile fracture.

  19. Laser shock processing to improve residual stresses with and without paint layer on 6061-T6 aluminum alloy

    NASA Astrophysics Data System (ADS)

    Gomez-Rosas, G.; Rubio-Gonzalez, C.; Ocaña, J. L.; Molpeceres, C.; Porro, J. A.; Morales, M.; Casillas, F. J.; Mora-Gonzalez, M.; Chi-Moreno, W.

    2007-03-01

    Laser shock processing (LSP) or laser shock peening has been proposed as a competitive alternative technology to classical treatments for improving fatigue and wear resistance of metals. This process induces a compressive residual stress field which increases fatigue crack initiation life and reduce fatigue crack growth rate. We present a configuration and results in the LSP concept for metal surface treatments in underwater laser irradiation at 1064 nm with and without a thin surface paint layer. A convergent lens is used to deliver 1, 2 J/cm2 with a 8 ns laser FWHM pulse produced by a 10 Hz, Q-switched Nd:YAG laser with a spot diameter of a 1,5 mm moving forward along the workpiece. A LSP configuration with experimental results using a pulse density of 5000 pulses/cm2 in 6061-T6 aluminum samples are presented.

  20. Effect of T6 heat treatment on the microstructural and mechanical properties of Al-Si-Cu-Mg alloys

    NASA Astrophysics Data System (ADS)

    Patel, Dhruv; Davda, Chintan; Solanki, P. S.; Keshvani, M. J.

    2016-05-01

    In this communication, it is aimed to optimize the conditions for T6 heat treatment of permanent die cast Al-Si-Cu-Mg alloys. Various solutionizing temperatures, aging treatments and soaking times were used to improve / modify the mechanical properties of presently studied alloys. Formation mechanism of the particles was understood by carrying out optical microscopy and energy dispersive X-ray (EDX) spectroscopy measurements. Spherical particles of alloys were studied for their microstructural properties using scanning electron microscopy (SEM). Microhardness test was performed to investigate their mechanical properties. Dependence of cluster formation and microhardness of the alloys on the adequate solutionizing temperature, aging treatment and soaking time has been discussed in detail.

  1. Crystallization and preliminary crystallographic analysis of a family 43 β-d-xylosidase from Geobacillus stearothermophilus T-6

    SciTech Connect

    Brüx, Christian; Niefind, Karsten; Ben-David, Alon; Leon, Maya; Shoham, Gil; Shoham, Yuval; Schomburg, Dietmar

    2005-12-01

    The crystallization and preliminary X-ray analysis of a β-d-xylosidase from G. stearothermophilus T-6, a family 43 glycoside hydrolase, is described. Native and catalytic inactive mutants of the enzymes were crystallized in two different space groups, orthorhombic P2{sub 1}2{sub 1}2 and tetragonal P4{sub 1}2{sub 1}2 (or the enantiomorphic space group P4{sub 3}2{sub 1}2), using a sensitive cryoprotocol. The latter crystal form diffracted X-rays to a resolution of 2.2 Å. β-d-Xylosidases (EC 3.2.1.37) are hemicellulases that cleave single xylose units from the nonreducing end of xylooligomers. In this study, the crystallization and preliminary X-ray analysis of a β-d-xylosidase from Geobacillus stearothermophilus T-6 (XynB3), a family 43 glycoside hydrolase, is described. XynB3 is a 535-amino-acid protein with a calculated molecular weight of 61 891 Da. Purified recombinant native and catalytic inactive mutant proteins were crystallized and cocrystallized with xylobiose in two different space groups, P2{sub 1}2{sub 1}2 (unit-cell parameters a = 98.32, b = 99.36, c = 258.64 Å) and P4{sub 1}2{sub 1}2 (or the enantiomorphic space group P4{sub 3}2{sub 1}2; unit-cell parameters a = b = 140.15, c = 233.11 Å), depending on the detergent. Transferring crystals to cryoconditions required a very careful protocol. Orthorhombic crystals diffract to 2.5 Å and tetragonal crystals to 2.2 Å.

  2. ATGL and CGI-58 are lipid droplet proteins of the hepatic stellate cell line HSC-T6.

    PubMed

    Eichmann, Thomas O; Grumet, Lukas; Taschler, Ulrike; Hartler, Jürgen; Heier, Christoph; Woblistin, Aaron; Pajed, Laura; Kollroser, Manfred; Rechberger, Gerald; Thallinger, Gerhard G; Zechner, Rudolf; Haemmerle, Günter; Zimmermann, Robert; Lass, Achim

    2015-10-01

    Lipid droplets (LDs) of hepatic stellate cells (HSCs) contain large amounts of vitamin A [in the form of retinyl esters (REs)] as well as other neutral lipids such as TGs. During times of insufficient vitamin A availability, RE stores are mobilized to ensure a constant supply to the body. To date, little is known about the enzymes responsible for the hydrolysis of neutral lipid esters, in particular of REs, in HSCs. In this study, we aimed to identify LD-associated neutral lipid hydrolases by a proteomic approach using the rat stellate cell line HSC-T6. First, we loaded cells with retinol and FAs to promote lipid synthesis and deposition within LDs. Then, LDs were isolated and lipid composition and the LD proteome were analyzed. Among other proteins, we found perilipin 2, adipose TG lipase (ATGL), and comparative gene identification-58 (CGI-58), known and established LD proteins. Bioinformatic search of the LD proteome for α/β-hydrolase fold-containing proteins revealed no yet uncharacterized neutral lipid hydrolases. In in vitro activity assays, we show that rat (r)ATGL, coactivated by rat (r)CGI-58, efficiently hydrolyzes TGs and REs. These findings suggest that rATGL and rCGI-58 are LD-resident proteins in HSCs and participate in the mobilization of both REs and TGs.

  3. ATGL and CGI-58 are lipid droplet proteins of the hepatic stellate cell line HSC-T6[S

    PubMed Central

    Eichmann, Thomas O.; Grumet, Lukas; Taschler, Ulrike; Hartler, Jürgen; Heier, Christoph; Woblistin, Aaron; Pajed, Laura; Kollroser, Manfred; Rechberger, Gerald; Thallinger, Gerhard G.; Zechner, Rudolf; Haemmerle, Günter; Zimmermann, Robert; Lass, Achim

    2015-01-01

    Lipid droplets (LDs) of hepatic stellate cells (HSCs) contain large amounts of vitamin A [in the form of retinyl esters (REs)] as well as other neutral lipids such as TGs. During times of insufficient vitamin A availability, RE stores are mobilized to ensure a constant supply to the body. To date, little is known about the enzymes responsible for the hydrolysis of neutral lipid esters, in particular of REs, in HSCs. In this study, we aimed to identify LD-associated neutral lipid hydrolases by a proteomic approach using the rat stellate cell line HSC-T6. First, we loaded cells with retinol and FAs to promote lipid synthesis and deposition within LDs. Then, LDs were isolated and lipid composition and the LD proteome were analyzed. Among other proteins, we found perilipin 2, adipose TG lipase (ATGL), and comparative gene identification-58 (CGI-58), known and established LD proteins. Bioinformatic search of the LD proteome for α/β-hydrolase fold-containing proteins revealed no yet uncharacterized neutral lipid hydrolases. In in vitro activity assays, we show that rat (r)ATGL, coactivated by rat (r)CGI-58, efficiently hydrolyzes TGs and REs. These findings suggest that rATGL and rCGI-58 are LD-resident proteins in HSCs and participate in the mobilization of both REs and TGs. PMID:26330055

  4. An investigation of the reduction in tensile strength and fatigue life of pre-corroded 7075-T6 aluminum alloy

    NASA Astrophysics Data System (ADS)

    Obert, B.; Ngo, K.; Hashemi, J.; Ekwaro-Osire, S.; Sivam, T. P.

    2000-08-01

    In aging aircraft, the synergetic interaction between corrosion and fatigue has been shown to reduce the life expectancy of aluminum alloys. The objective of this study was to quantify the effects of corrosion, in terms of mass loss per unit area, on the static strength and fatigue life of 7075-T6 aluminum alloy. This was an experimental study in which test specimens were corroded in a laboratory environment. The corrosion process was accelerated by use of a corrosion cell. Test specimens were cut from flat sheets of aluminum and covered with masking material to restrict corrosion to a confined area. After testing, the fatigue life, ultimate tensile strength (UTS), and hardness of the specimens were observed to drop significantly with small amounts of corrosion. After the initial decrease, the UTS was observed to decrease linearly with increasing corrosion levels. The fatigue life of the specimens decreased in an inverse exponential fashion as mass loss per unit area increased. The hardness values of the corroded surfaces were also observed to drop. The topology of the pits and the related subsurface damage produced areas of high stress concentration resulting in the immediate reduction of UTS and fatigue life of the specimens. Subsurface corrosion damage was responsible for the reduction in hardness.

  5. A study on ultra-precision machining technique for Al6061-T6 to fabricate space infrared optics

    NASA Astrophysics Data System (ADS)

    Ryu, Geun-man; Lee, Gil-jae; Hyun, Sang-won; Sung, Ha-yeong; Chung, Euisik; Kim, Geon-hee

    2014-08-01

    In this paper, analysis of variance on designed experiments with full factorial design was applied to determine the optimized machining parameters for ultra-precision fabrication of the secondary aspheric mirror, which is one of the key elements of the space cryogenic infrared optics. A single point diamond turning machine (SPDTM, Nanotech 4μpL Moore) was adopted to fabricate the material, AL6061-T6, and the three machining parameters of cutting speed, feed rate and depth of cut were selected. With several randomly assigned experimental conditions, surface roughness of each condition was measured by a non-contact optical profiler (NT2000; Vecco). As a result of analysis using Minitab, the optimum cutting condition was determined as following; cutting speed: 122 m/min, feed rate: 3 mm/min and depth of cut: 1 μm. Finally, a 120 mm diameter aspheric secondary mirror was attached to a particularly designed jig by using mixture of paraffin and wax and successfully fabricated under the optimum machining parameters. The profile of machined surface was measured by a high-accuracy 3-D profilometer(UA3P; Panasonic) and we obtained the geometrical errors of 30.6 nm(RMS) and 262.4 nm(PV), which satisfy the requirements of the space cryogenic infrared optics.

  6. Surface analytical study of the corrosion behaviour of chromate passivated Al 2014 A T-6 during salt fog exposure

    NASA Astrophysics Data System (ADS)

    Treacy, G. M.; Wilcox, G. D.

    2000-03-01

    Surface analysis, using X-ray photoelectron spectroscopy (XPS), Auger electron spectroscopy (AES), and laser induced mass analysis (LIMA), was carried out on chromate conversion coated aluminium alloy 2014 A T-6 prior to, and after, exposure to a neutral sodium chloride salt fog environment. From XPS, no chromium could be detected at these surfaces after a 48-h salt fog exposure. AES also indicated that the chromium content decreased after a 24-h salt fog exposure. However, from depth profiling, carried out using AES prior to and after a 24-h salt fog exposure, the chromium concentration was observed to increase within a few nanometres of the surface, and appeared unaffected by the exposure to the corrosive salt fog environment. During exposure, dark stains were observed to develop on these surfaces within 48 h. LIMA, carried out on these stained areas, indicated the presence of Fe, CN and Cu species, as well as aluminium hydroxide species, indicating that this initial coating failure may occur at copper-containing intermetallic sites on these surfaces. It would appear that the appearance of black stains on these surfaces is the first indication of coating failure, rather than the development of white corrosion product (WCP), as the appearance of such stains coincided with the loss of chromate species from the surface.

  7. Ultrasonic Fatigue Endurance of Aluminum Alloy AISI 6061-T6 on Pre-corroded and Non-corroded Specimens

    NASA Astrophysics Data System (ADS)

    Domínguez Almaraz, Gonzalo M.; Dueñas Aburto, Antonio; Correa Gómez, Erasmo

    2014-01-01

    Ultrasonic fatigue tests are carried out on aluminum alloy 6061-T6 in order to analyze the fatigue endurance behavior under artificial pre-corrosion attack by hydrochloric acid for the pH concentrations of 0.47 and 0.80. The pre-corrosion attack is used to simulate the long-time environmental effect and the corresponding decay of fatigue life in regard to non-corroded specimens. Experimental results show that ultrasonic fatigue endurance under these two degrees of pre-corrosion attack decreases dramatically. Furthermore, it is observed that crack initiation is frequently associated with one or several pre-corrosion pitting holes at the specimen surface. Pitting holes are assumed to be semi-hemispherical and the stress concentration factors are evaluated taking into account the size and proximity of two crack initiation pitting holes. The crack growth rates are obtained for the pre-corroded specimens and compared to the non-corroded specimen. Finally, conclusions are listed concerning ultrasonic fatigue endurance of testing specimens, together with the fracture surfaces, crack paths, and crack growth rates.

  8. ATGL and CGI-58 are lipid droplet proteins of the hepatic stellate cell line HSC-T6.

    PubMed

    Eichmann, Thomas O; Grumet, Lukas; Taschler, Ulrike; Hartler, Jürgen; Heier, Christoph; Woblistin, Aaron; Pajed, Laura; Kollroser, Manfred; Rechberger, Gerald; Thallinger, Gerhard G; Zechner, Rudolf; Haemmerle, Günter; Zimmermann, Robert; Lass, Achim

    2015-10-01

    Lipid droplets (LDs) of hepatic stellate cells (HSCs) contain large amounts of vitamin A [in the form of retinyl esters (REs)] as well as other neutral lipids such as TGs. During times of insufficient vitamin A availability, RE stores are mobilized to ensure a constant supply to the body. To date, little is known about the enzymes responsible for the hydrolysis of neutral lipid esters, in particular of REs, in HSCs. In this study, we aimed to identify LD-associated neutral lipid hydrolases by a proteomic approach using the rat stellate cell line HSC-T6. First, we loaded cells with retinol and FAs to promote lipid synthesis and deposition within LDs. Then, LDs were isolated and lipid composition and the LD proteome were analyzed. Among other proteins, we found perilipin 2, adipose TG lipase (ATGL), and comparative gene identification-58 (CGI-58), known and established LD proteins. Bioinformatic search of the LD proteome for α/β-hydrolase fold-containing proteins revealed no yet uncharacterized neutral lipid hydrolases. In in vitro activity assays, we show that rat (r)ATGL, coactivated by rat (r)CGI-58, efficiently hydrolyzes TGs and REs. These findings suggest that rATGL and rCGI-58 are LD-resident proteins in HSCs and participate in the mobilization of both REs and TGs. PMID:26330055

  9. Plastic Stress-strain Relations for 75S-T6 Aluminum Alloy Subjected to Biaxial Tensile Stresses

    NASA Technical Reports Server (NTRS)

    Marin, Joseph; Ulrich, B H; Hughes, W P

    1951-01-01

    In this investigation, the material tested was a 75S-T6 aluminum alloy and the stresses were essentially biaxial and tensile. The biaxial tensile stresses were produced in a specially designed testing machine by subjecting a thin-walled tubular specimen to axial tension and internal pressure. Plastic stress-strain relations for various biaxial stress conditions were obtained using a clip-type SR-4 strain gage. Three types of tests were made: Constant-stress-ratio tests, variable-stress-ratio tests, and special tests. The constant-stress-ratio test results gave control data and showed the influence of biaxial stresses on the yield, fracture, and ultimate strength of the material. By means of the variable-stress-ratio tests, it is possible to determine whether there is any significant difference between the flow and deformation type of theory. Finally, special tests were conducted to check specific assumptions made in the theories of plastic flow. The constant-stress-ratio tests show that the deformation theory based on the octahedral, effective; or significant stress-strain relations is in approximate agreement with the test results. The variable-stress-ratio tests show that both the deformation and flow theory are in equally good agreement with the test results.

  10. On Residual Stresses in Resistance Spot-Welded Aluminum Alloy 6061-T6: Experimental and Numerical Analysis

    NASA Astrophysics Data System (ADS)

    Afshari, D.; Sedighi, M.; Karimi, M. R.; Barsoum, Z.

    2013-12-01

    In this study, an electro-thermal-structural-coupled finite element (FE) model and x-ray diffraction residual stress measurements have been utilized to analyze distribution of residual stresses in an aluminum alloy 6061-T6 resistance spot-welded joint with 2-mm-thickness sheet. Increasing the aluminum sheet thickness to more than 1 mm leads to creating difficulty in spot-welding process and increases the complexity of the FE model. The electrical and thermal contact conductances, as mandatory factors are applied in contact areas of electrode-workpiece and workpiece-workpiece to resolve the complexity of the FE model. The physical and mechanical properties of the material are defined as thermal dependent to improve the accuracy of the model. Furthermore, the electrodes are removed after the holding cycle using the birth-and-death elements method. The results have a good agreement with experimental data obtained from x-ray diffraction residual stress measurements. However, the highest internal tensile residual stress occurs in the center of the nugget zone and decreases toward nugget edge; surface residual stress increases toward the edge of the welding zone and afterward, the area decreases slightly.

  11. Crystallization and preliminary crystallographic analysis of a family 43 β-d-xylosidase from Geobacillus stearothermophilus T-6

    PubMed Central

    Brüx, Christian; Niefind, Karsten; Ben-David, Alon; Leon, Maya; Shoham, Gil; Shoham, Yuval; Schomburg, Dietmar

    2005-01-01

    β-d-Xylosidases (EC 3.2.1.37) are hemicellulases that cleave single xylose units from the nonreducing end of xylooligomers. In this study, the crystallization and preliminary X-ray analysis of a β-d-xylosidase from Geobacillus stearothermophilus T-6 (XynB3), a family 43 glycoside hydrolase, is described. XynB3 is a 535-amino-acid protein with a calculated molecular weight of 61 891 Da. Purified recombinant native and catalytic inactive mutant proteins were crystallized and cocrystallized with xylobiose in two different space groups, P21212 (unit-cell parameters a = 98.32, b = 99.36, c = 258.64 Å) and P41212 (or the enantiomorphic space group P43212; unit-cell parameters a = b = 140.15, c = 233.11 Å), depending on the detergent. Transferring crystals to cryoconditions required a very careful protocol. Orthorhombic crystals diffract to 2.5 Å and tetragonal crystals to 2.2 Å. PMID:16511233

  12. t(6;11) renal cell carcinoma (RCC): expanded immunohistochemical profile emphasizing novel RCC markers and report of 10 new genetically confirmed cases.

    PubMed

    Smith, Nathaniel E; Illei, Peter B; Allaf, Mohamed; Gonzalez, Nilda; Morris, Kerry; Hicks, Jessica; Demarzo, Angelo; Reuter, Victor E; Amin, Mahul B; Epstein, Jonathan I; Netto, George J; Argani, Pedram

    2014-05-01

    Renal cell carcinomas (RCCs) harboring the t(6;11)(p21;q12) translocation were first described in 2001 and recently recognized by the 2013 International Society of Urological Pathology Vancouver Classification of Renal Neoplasia. Although these RCCs are known to label for melanocytic markers HMB45 and Melan A and the cysteine protease cathepsin K by immunohistochemistry (IHC), a comprehensive IHC profile has not been reported. We report 10 new t(6;11) RCCs, all confirmed by break-apart TFEB fluorescence in situ hybridization. A tissue microarray containing 6 of these cases and 7 other previously reported t(6;11) RCCs was constructed and immunolabeled for 21 different antigens. Additional whole sections of t(6;11) RCC were labeled with selected IHC markers. t(6;11) RCC labeled diffusely and consistently for cathepsin K and Melan A (13 of 13 cases) and almost always at least focally for HMB45 (12 of 13 cases). They labeled frequently for PAX8 (14 of 23 cases), CD117 (10 of 14 cases), and vimentin (9 of 13 cases). A majority of cases labeled at least focally for cytokeratin Cam5.2 (8 of 13 cases) and CD10 and RCC marker antigen (10 of 14 cases each). In contrast to a prior study's findings, only a minority of cases labeled for Ksp-cadherin (3 of 19 cases). The median H score (product of intensity score and percentage labeling) for phosphorylated S6, a marker of mTOR pathway activation, was 101, which is high relative to most other RCC subtypes. In summary, IHC labeling for PAX8, Cam5.2, CD10, and RCC marker antigen supports classification of the t(6;11) RCC as carcinomas despite frequent negativity for broad-spectrum cytokeratins and EMA. Labeling for PAX8 distinguishes the t(6;11) RCC from epithelioid angiomyolipoma, which otherwise shares a similar immunoprofile. CD117 labeling is more frequent in the t(6;11) RCC compared with the related Xp11 translocation RCC. Increased pS6 expression suggests a possible molecular target for the uncommon t(6;11) RCCs that

  13. Effect of caffeine on motility and vitality of sperm and in vitro fertilization of outbreed mouse in T6 and M16 media

    PubMed Central

    Nabavi, Narges; Todehdehghan, Fatemeh; Shiravi, Abdollhossein

    2013-01-01

    Background: Caffeine increases the CAMP production that stimulates spermatozoa movement. Caffeine is also used for induction of in vitro acrosome reaction in mammalian spermatozoa, an important step in achieving fertilization. Objective: The aim of this study was to assess the effect of caffeine on sperm's motility, vitality and laboratory fertilization rates in mouse in two T6 and M16 media. Materials and Methods: Epididymal mouse sperms were collected and treated by caffine in T6 and M16 media and their motility and vitality rates were evaluated. The pretreated sperms were added to oocytes in T6 and M16 media with and without caffeine and fertilization rates were recorded after 24 hours incubation. Results: Sperm's motility (81.7±1.67%) and vitality (88.7±1.33%) rates and percentage of fertilized oocytes (67.52±8.16%) in T6 medium plus caffeine compare to control group have increased and shown significant differences at p≤0.01. While the percentages of these parameters in M16 medium supplemented with caffeine were 68.3±6.01%, 78±6.11%, and 42.6±12.96 respectively and in comparison to control group (M16 without caffeine) have not shown significant differences. Conclusion: Addition of caffeine to T6 medium promotes the sperm's motility and vitality and enhances fertilization and early in vitro development of mouse embryos. This article extracted from M.Sc. thesis. (Narges Navabi) PMID:24639814

  14. Murine interferon system regulation: isolation and characterization of a mutant 3T6 cell engaged in the semiconstitutive synthesis of interferon.

    PubMed

    Jarvis, A P; Colby, C

    1978-06-01

    We describe the isolation and characterization of a virus-resistant mutant of murine 3T6 cells. The mutant, designated 3T6-VrB2, displays a high degree of resistance to infection by members of the toga-, rhabdo- and picornavirus classes. The level of this resistance to infection is similar to the parent 3T6 pretreated with approximately 100 lU/ml of interferon. Upon co-cultivation of 3T6-VrB2 cells with interferon-sensitive mouse cells, an antiviral state is induced in the latter cells as measured by a reduction of virus yield following infection. The nature of the induction is defined by a series of experiments using anti-mouse interferon antiserum. In the presence of this antiserum, the ability of the mutant to induce an antiviral state in interferon-sensitive mouse cells upon co-cultivation is eliminated. Additionally, growth of the mutant cells in the presence of this antiserum causes a reversal of the virus-resistant phenotype. Our results indicate that 3T6-VrB2 contains a mutation affecting the regulation of the murine interferon system such that the cell is engaged in the semiconstitutive synthesis of interferon. PMID:208779

  15. Barrier anodic coatings formed on 6061-T6 aluminum alloy in electrolytes containing different ethanol to water ratios

    SciTech Connect

    Panitz, J.K.G.; Sharp, D.J.; Martinez, F.E.; Merrill, R.M.; Ward, K.J.

    1988-12-01

    We have studied barrier anodic film formation on 6061-T6 aluminum alloy substrates as a function of electrolyte composition for five mixtures of ammonium tartrate dissolved in water and diluted with different amounts of ethanol. The effects of electrolyte temperatures within the range of 18/degree/C to 38/degree/C were explored. The results of this study indicate that the best dielectric coatings and the shortest processing times occur for the 100% water-ammonium tartrate electrolyte. The second best coatings and processing times occur in conjunction with the use of 98% ethanol, 2% water plus ammonium tartrate electrolyte. In general, visibly flawed coatings, scintillation events at cell voltages in excess of approximately 750-800 volts and/or abnormally long processing times occur in conjunction with the use of electrolyte mixtures containing 20%, 60%, and 90% water. We analysed samples of electrolyte as a function of usage, and evaluated the composition of the coatings using Fourier Transform Infrared Analysis to better understand the mechanisms which contribute to anodic coating growth that result in the observed variations in the dielectric properties. All of the coatings exhibited similar compositions except with regard to the amount of CO2 that was physisorbed in the coatings. The dielectrically inferior coatings that were typically produced by the electrolytes containing ethanol contain substantially more CO2 than the coatings grown in the 100% water-based electrolyte. These results strongly suggest that the ethanol in the electrolyte oxidizes and forms CO2 which is incorporated in the coatings and results in inferior dielectric properties. 8 refs., 7 figs.

  16. Artificial neural networks application for modeling of friction stir welding effects on mechanical properties of 7075-T6 aluminum alloy

    NASA Astrophysics Data System (ADS)

    Maleki, E.

    2015-12-01

    Friction stir welding (FSW) is a relatively new solid-state joining technique that is widely adopted in manufacturing and industry fields to join different metallic alloys that are hard to weld by conventional fusion welding. Friction stir welding is a very complex process comprising several highly coupled physical phenomena. The complex geometry of some kinds of joints makes it difficult to develop an overall governing equations system for theoretical behavior analyse of the friction stir welded joints. Weld quality is predominantly affected by welding effective parameters, and the experiments are often time consuming and costly. On the other hand, employing artificial intelligence (AI) systems such as artificial neural networks (ANNs) as an efficient approach to solve the science and engineering problems is considerable. In present study modeling of FSW effective parameters by ANNs is investigated. To train the networks, experimental test results on thirty AA-7075-T6 specimens are considered, and the networks are developed based on back propagation (BP) algorithm. ANNs testing are carried out using different experimental data that they are not used during networks training. In this paper, rotational speed of tool, welding speed, axial force, shoulder diameter, pin diameter and tool hardness are regarded as inputs of the ANNs. Yield strength, tensile strength, notch-tensile strength and hardness of welding zone are gathered as outputs of neural networks. According to the obtained results, predicted values for the hardness of welding zone, yield strength, tensile strength and notch-tensile strength have the least mean relative error (MRE), respectively. Comparison of the predicted and the experimental results confirms that the networks are adjusted carefully, and the ANN can be used for modeling of FSW effective parameters.

  17. Preliminary crystallographic analysis of Xyn52B2, a GH52 β-D-xylosidase from Geobacillus stearothermophilus T6.

    PubMed

    Dann, Roie; Lansky, Shifra; Lavid, Noa; Zehavi, Arie; Belakhov, Valery; Baasov, Timor; Dvir, Hay; Manjasetty, Babu; Belrhali, Hassan; Shoham, Yuval; Shoham, Gil

    2014-12-01

    Geobacillus stearothermophilus T6 is a thermophilic bacterium that possesses an extensive hemicellulolytic system, including over 40 specific genes that are dedicated to this purpose. For the utilization of xylan, the bacterium uses an extracellular xylanase which degrades xylan to decorated xylo-oligomers that are imported into the cell. These oligomers are hydrolyzed by side-chain-cleaving enzymes such as arabinofuranosidases, acetylesterases and a glucuronidase, and finally by an intracellular xylanase and a number of β-xylosidases. One of these β-xylosidases is Xyn52B2, a GH52 enzyme that has already proved to be useful for various glycosynthesis applications. In addition to its demonstrated glycosynthase properties, interest in the structural aspects of Xyn52B2 stems from its special glycoside hydrolase family, GH52, the structures and mechanisms of which are only starting to be resolved. Here, the cloning, overexpression, purification and crystallization of Xyn52B2 are reported. The most suitable crystal form that has been obtained belonged to the orthorhombic P212121 space group, with average unit-cell parameters a = 97.7, b = 119.1, c = 242.3 Å. Several X-ray diffraction data sets have been collected from flash-cooled crystals of this form, including the wild-type enzyme (3.70 Å resolution), the E335G catalytic mutant (2.95 Å resolution), a potential mercury derivative (2.15 Å resolution) and a selenomethionine derivative (3.90 Å resolution). These data are currently being used for detailed three-dimensional structure determination of the Xyn52B2 protein.

  18. Preliminary crystallographic analysis of Xyn52B2, a GH52 β-D-xylosidase from Geobacillus stearothermophilus T6.

    PubMed

    Dann, Roie; Lansky, Shifra; Lavid, Noa; Zehavi, Arie; Belakhov, Valery; Baasov, Timor; Dvir, Hay; Manjasetty, Babu; Belrhali, Hassan; Shoham, Yuval; Shoham, Gil

    2014-12-01

    Geobacillus stearothermophilus T6 is a thermophilic bacterium that possesses an extensive hemicellulolytic system, including over 40 specific genes that are dedicated to this purpose. For the utilization of xylan, the bacterium uses an extracellular xylanase which degrades xylan to decorated xylo-oligomers that are imported into the cell. These oligomers are hydrolyzed by side-chain-cleaving enzymes such as arabinofuranosidases, acetylesterases and a glucuronidase, and finally by an intracellular xylanase and a number of β-xylosidases. One of these β-xylosidases is Xyn52B2, a GH52 enzyme that has already proved to be useful for various glycosynthesis applications. In addition to its demonstrated glycosynthase properties, interest in the structural aspects of Xyn52B2 stems from its special glycoside hydrolase family, GH52, the structures and mechanisms of which are only starting to be resolved. Here, the cloning, overexpression, purification and crystallization of Xyn52B2 are reported. The most suitable crystal form that has been obtained belonged to the orthorhombic P212121 space group, with average unit-cell parameters a = 97.7, b = 119.1, c = 242.3 Å. Several X-ray diffraction data sets have been collected from flash-cooled crystals of this form, including the wild-type enzyme (3.70 Å resolution), the E335G catalytic mutant (2.95 Å resolution), a potential mercury derivative (2.15 Å resolution) and a selenomethionine derivative (3.90 Å resolution). These data are currently being used for detailed three-dimensional structure determination of the Xyn52B2 protein. PMID:25484225

  19. Ballistic limit of 6061 T6 aluminum and threat to surface coatings for use with orbiting space station space suit materials

    NASA Technical Reports Server (NTRS)

    Fish, D.

    1986-01-01

    In recent years orbiting satellites, spent components, collisions and explosions have populated the near earth orbit with debris potentially more hazardous than the average meteoroid debris. This new debris has an average density of aluminum (2.78 g/cc) and an average encounter velocity of 10 km/sec. The space station will require many hours of work in this environment and there is concern over hazard to the assembly personnel. A proposed hard suit design utilizes 6061-T6 Aluminum for most of its exposed area. The aluminum surface will be treated for thermal and radiation control. The basic thickness of this suit will be on the order of 1.78 mm (0.070 inches). The selection of 6061-T6 Aluminum for space suits for use on the space station would appear to be worthwhile. The relatively ductile behavior of 6061-T6 aluminum is better than a choice of a more brittle material.

  20. New plant growth-modifying properties of the Agrobacterium T-6b oncogene revealed by the use of a dexamethasone-inducible promoter.

    PubMed

    Grémillon, Louis; Helfer, Anne; Clément, Bernadette; Otten, Léon

    2004-01-01

    Agrobacterium 6b oncogenes induce tumours on Nicotiana glauca and enations and associated modifications in transgenic N. tabacum plants. 2x35S-AB-6b tobacco rootstocks produced a graft-transmissible factor that induced enations in wild-type scions; the nature of this enation factor remains to be identified. Here, we report on the properties of tobacco plants carrying a dexamethasone-inducible T-6b gene (dex-T-6b). Induction with dex led to complex growth modifications, many of which have not been reported previously. Modifications were only found in growing tissues; mature tissues remained unaffected. Growth could be either stimulated or inhibited. Dex induction of young plants led to morphogenetic gradients that included enations, tubular leaves and fragmented leaf primordia. Root elongation was increased or slowed down, while radial root growth was strongly enhanced. Additional cell divisions were found in the root pericycle and vasculature. Enation factor import from mature tissues did not have the same effects on growing tissues as local T-6b synthesis: normal scions grafted on induced dex-T-6b rootstocks formed enations, whereas local dex-T-6b induction at the shoot apex led to numerous dark-green spots on the abaxial side of the leaves. In leaf patch assays, the 23-kDa T-6b protein was found to move through leaves and to enter the vascular system. This and the fact that rootstocks of spontaneous tobacco enation mutants did not modify wild-type scions contrary to 6b plants indicate that the 6b protein might be the enation factor. PMID:14690506

  1. H-NS binding to evpB and evpC and repressing T6SS expression in fish pathogen Edwardsiella piscicida.

    PubMed

    Cui, Shilei; Xiao, Jingfan; Wang, Qiyao; Zhang, Yuanxing

    2016-09-01

    Edwardsiella piscicida is an important causative agent of hemorrhagic septicemia in fish and infects both cultured and wild fish species. Type VI secretion system (T6SS) was proved to play important roles in pathogenesis of E. piscicida. In this study, it was demonstrated that the expression of T6SS genes evpB and evpC was under control of the global regulator H-NS in E. piscicida and the transcriptional level of evpB and evpC was significantly down-regulated by H-NS. Compared to the wild type, the transcriptional levels of evpB and evpC were up-regulated in hns null mutant, while down-regulated in hns overexpression strain. The results of EMSA and DNase I footprinting revealed that H-NS protein directly bound to upstream region of evpC at multiple sites. A high-affinity motif with a 9-nucleotide sequence 5'-ATATAAAAT-3' was defined for H-NS preferential recognition based on the feature of the binding sites. These results indicated that H-NS acted cooperatively to form extended nucleoprotein filaments on target DNA. Site-directed mutagenesis of H-NS further showed that R86 played an essential role in T6SS gene binding. These findings highlighted the mechanisms underlying the complex regulation network of T6SS by H-NS in E. piscicida.

  2. Structure-specificity relationships in Abp, a GH27 β-L-arabinopyranosidase from Geobacillus stearothermophilus T6.

    PubMed

    Lansky, Shifra; Salama, Rachel; Solomon, Hodaya V; Feinberg, Hadar; Belrhali, Hassan; Shoham, Yuval; Shoham, Gil

    2014-11-01

    L-Arabinose sugar residues are relatively abundant in plants and are found mainly in arabinan polysaccharides and in other arabinose-containing polysaccharides such as arabinoxylans and pectic arabinogalactans. The majority of the arabinose units in plants are present in the furanose form and only a small fraction of them are present in the pyranose form. The L-arabinan-utilization system in Geobacillus stearothermophilus T6, a Gram-positive thermophilic soil bacterium, has recently been characterized, and one of the key enzymes was found to be an intracellular β-L-arabinopyranosidase (Abp). Abp, a GH27 enzyme, was shown to remove β-L-arabinopyranose residues from synthetic substrates and from the native substrates sugar beet arabinan and larch arabinogalactan. The Abp monomer is made up of 448 amino acids, and based on sequence homology it was suggested that Asp197 is the catalytic nucleophile and Asp255 is the catalytic acid/base. In the current study, the detailed three-dimensional structure of wild-type Abp (at 2.28 Å resolution) and its catalytic mutant Abp-D197A with (at 2.20 Å resolution) and without (at 2.30 Å resolution) a bound L-arabinose product are reported as determined by X-ray crystallography. These structures demonstrate that the three-dimensional structure of the Abp monomer correlates with the general fold observed for GH27 proteins, consisting of two main domains: an N-terminal TIM-barrel domain and a C-terminal all-β domain. The two catalytic residues are located in the TIM-barrel domain, such that their carboxylic functional groups are about 5.9 Å from each other, consistent with a retaining mechanism. An isoleucine residue (Ile67) located at a key position in the active site is shown to play a critical role in the substrate specificity of Abp, providing a structural basis for the high preference of the enzyme towards arabinopyranoside over galactopyranoside substrates. The crystal structure demonstrates that Abp is a tetramer

  3. Structure-specificity relationships in Abp, a GH27 β-L-arabinopyranosidase from Geobacillus stearothermophilus T6.

    PubMed

    Lansky, Shifra; Salama, Rachel; Solomon, Hodaya V; Feinberg, Hadar; Belrhali, Hassan; Shoham, Yuval; Shoham, Gil

    2014-11-01

    L-Arabinose sugar residues are relatively abundant in plants and are found mainly in arabinan polysaccharides and in other arabinose-containing polysaccharides such as arabinoxylans and pectic arabinogalactans. The majority of the arabinose units in plants are present in the furanose form and only a small fraction of them are present in the pyranose form. The L-arabinan-utilization system in Geobacillus stearothermophilus T6, a Gram-positive thermophilic soil bacterium, has recently been characterized, and one of the key enzymes was found to be an intracellular β-L-arabinopyranosidase (Abp). Abp, a GH27 enzyme, was shown to remove β-L-arabinopyranose residues from synthetic substrates and from the native substrates sugar beet arabinan and larch arabinogalactan. The Abp monomer is made up of 448 amino acids, and based on sequence homology it was suggested that Asp197 is the catalytic nucleophile and Asp255 is the catalytic acid/base. In the current study, the detailed three-dimensional structure of wild-type Abp (at 2.28 Å resolution) and its catalytic mutant Abp-D197A with (at 2.20 Å resolution) and without (at 2.30 Å resolution) a bound L-arabinose product are reported as determined by X-ray crystallography. These structures demonstrate that the three-dimensional structure of the Abp monomer correlates with the general fold observed for GH27 proteins, consisting of two main domains: an N-terminal TIM-barrel domain and a C-terminal all-β domain. The two catalytic residues are located in the TIM-barrel domain, such that their carboxylic functional groups are about 5.9 Å from each other, consistent with a retaining mechanism. An isoleucine residue (Ile67) located at a key position in the active site is shown to play a critical role in the substrate specificity of Abp, providing a structural basis for the high preference of the enzyme towards arabinopyranoside over galactopyranoside substrates. The crystal structure demonstrates that Abp is a tetramer

  4. Full Field Measurement of The Dynamic Response of AA6061-T6 Aluminum Alloy under High Strain Rate Compression and Torsion Loads

    NASA Astrophysics Data System (ADS)

    Odoh, Daniel Oghenekewhe Oluwatobi

    The dynamic response of AA6061-T6 aluminum alloy under high strain rate loading in compression and torsion loading conditions was studied using the split Hopkinson pressure bar, the Kolsky torsion bar, and the high speed digital image correlation system. AA6061-T6 alloy, the most widely used in the AA6000 series, is a multi-purpose Al-Mg-Si-Cu alloy containing about 0.4 % wt. of Cu and other alloying additives. The properties of AA6061-T6 aluminum alloy including medium to high strength, good fracture toughness, and high corrosion resistance make it to find application in high performance structures such as the automotive parts, panels, and armored carriers. In this work, the effect of strain rate during dynamic test on formation of adiabatic shear bands in AA6061-T6 alloy was investigated. A post deformation analysis of the tested specimen was performed in order to determine the damage evolution and strain localization along the narrow adiabatic shear bands within the specimen. The formation of an adiabatic shear band in the aluminum alloy tested was found to depend on the strain rate at which the test was conducted. Stress, strain, and strain rate data obtained from the elastic waves in the compression and torsion bar tests were also compared with those obtained using the high speed digital cameras. Results show good agreement between both measurement techniques with the 3D digital image correlation technique giving a slightly lower result. Scanning and electron microscopy results show that both deformed and transformed bands can be formed in AA6061-T6 alloy during dynamic loading. The type of adiabatic shear band formed depends on the strain rate at which test was performed.

  5. Direct-reading design charts for 75S-T6 aluminum-alloy flat compression panels having longitudinal extruded Z-section stiffeners

    NASA Technical Reports Server (NTRS)

    Hickman, William A; Dow, Norris F

    1951-01-01

    Direct-reading design charts are presented for 75S-T6 aluminum-alloy flat compression panels having longitudinal extruded Z-section stiffeners. These charts, which cover a wide range of proportions, make possible the direct determination of the stress and all panel dimensions required to carry a given intensity of loading with a given skin thickness and effective length of panel.

  6. Genomic analysis of the biocontrol strain Pseudomonas fluorescens Pf29Arp with evidence of T3SS and T6SS gene expression on plant roots.

    PubMed

    Marchi, Muriel; Boutin, Morgane; Gazengel, Kévin; Rispe, Claude; Gauthier, Jean-Pierre; Guillerm-Erckelboudt, Anne-Yvonne; Lebreton, Lionel; Barret, Matthieu; Daval, Stéphanie; Sarniguet, Alain

    2013-06-01

    Several bacterial strains of the Pseudomonas genus provide plant growth stimulation, plant protection against pests or bioremediation. Among these bacteria, P. fluorescens Pf29Arp reduces the severity of take-all, a disease caused by the pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) on wheat roots. In this study, we obtained a draft genome of Pf29Arp and subsequent comparative genomic analyses have revealed that this bacterial strain is closely related to strains of the 'P. brassicacearum-like' subgroup including P. brassicacearum ssp. brassicacearum NFM421 and P. fluorescens F113. Despite an overall chromosomal organization similar to these strains, a number of features including antibiotic synthesis gene clusters from secondary metabolism are not found in the Pf29Arp genome. But Pf29Arp possesses different protein secretion systems including type III (T3SS) and type VI (T6SS) secretion systems. Pf29Arp is the first Pseudomonas sp. strain described with four T6SS clusters (cluster I, II, III and IV). In addition, some protein-coding genes involved in the assembly of these secretion systems are basally expressed during Pf29Arp colonization of healthy wheat roots and display different expression patterns on necrotized roots caused by Ggt. These data suggest a role of T3SS and T6SS in the Pf29Arp adaptation to different root environments.

  7. Time exposure studies on stress corrosion cracking of aluminum 2014-T6, 2219-T87, 2014-T651, 7075-T651, and titanium 6Al-4V

    NASA Technical Reports Server (NTRS)

    Terrell, J.

    1973-01-01

    The effect of a constant applied stress in crack initiation of aluminum 2014-T6, 2219-T87, 2014-T651, 7075-T651 and titanium 6Al-4V has been investigated. Aluminum c-ring specimens (1-inch diameter) and u-band titanium samples were exposed continuously to a 3.5% NaCl solution (pH 7) and organic fluids of ethyl, methyl, and iso-propyl alcohol (reagent purity), and demineralized distilled water. Corrosive action was observed to begin during the first and second day of constant exposure as evidenced by accumulation of hydrogen bubbles on the surface of stressed aluminum samples. However, titanium stressed specimens showed no reactions to its environment. Results of this investigation seems to suggest that aluminum 2014-T6, aluminum 7075-T651 and aluminum 2014-T651 are susceptible to stress corrosion cracking in chloride solution (NaCl), while aluminum 2219-T87 seem to resist stress corrosion cracking in sodium chloride at three levels of stress (25%, 50%, and 75% Y.S.). In organic fluids of methyl, ethyl, and iso-propyl alcohol, 2014-T6 and 7075-T651 did not fail by SCC; but 2014-T651 was susceptible to SCC in methly alcohol, but resistant in ethyl alcohol, iso-propyl alcohol and demineralized distilled water.

  8. Galvanic coupling between D6AC steel, 6061-T6 aluminum, Inconel 718 and graphite-epoxy composite material: Corrosion occurrence and prevention

    NASA Technical Reports Server (NTRS)

    Danford, M. D.; Higgins, R. H.

    1983-01-01

    The effects of galvanic coupling between D6AC steel, 6061-T6 aluminum, Inconel 718, and graphite-epoxy composite material (G/E) in 3.5% NaCl were studied. Measurements of corrosion potentials, galvanic currents and corrosion rates of the bare metals using weight-loss methods served to establish the need for corrosion protection in cases where D6AC steel and 6061-T6 aluminum are galvanically coupled to G/E in salt water while Inconel 718 was shown to be compatible with G/E. Six tests were made to study corrosion protective methods for eliminating galvanic corrosion in the cases of D6AC steel and 6061-T6 aluminum coupled to G/E. These results indicate that, when the G/E is completely coated with paint or a paint/polyurethane resin combination, satisfactory protection of the D6AC steel is achieved with either a coat of zinc-rich primer or a primer/topcoat combination. Likewise, satisfactory corrosion protection of the aluminum is achieved by coating it with an epoxy coating system.

  9. Testing of the Qinetiq T6 Thruster in Support of the ESA BepiColombo Mercury Mission for the ESA BepiColombo Mission

    NASA Astrophysics Data System (ADS)

    Wallace, N.

    2004-10-01

    This paper presents the results of two test programmes. The first involved a series of simultaneous thruster firing tests performed at QinetiQ in the QinetiQ LEEP 2 (2nd Large European Electric Propulsion) vacuum test facility using 2 T6 thrusters operating between 50 and 230 mN at a specific impulse above 4500s. The test configuration allowed the simultaneous operation of 2 thrusters at a range of thrust levels and thruster spacings. The spacing between thrusters could be varied between 800 mm and 10 mm with the thrusters operating. One of the thrusters, designated the Primary, was suspended from a pendulum thrust balance and was aligned with a large diameter ion beam probe array. These diagnostics allowed the thrust to be directly measured and the ion beam plume to be characterised, allowing comparisons to be made between single and simultaneous operation at different thruster spacings. Testing was also performed with different combinations of neutralisers to determine the potential effects of dual thruster operation with a single neutraliser. The second series of tests involved the operation of a single T6 thruster at a thrust level of 208 mN at the elevated thermal conditions predicted for Mercury. Thermal and mechanical analyses of the T6 (including additional thrusters in close proximity) indicated the worst case thermal conditions occur when the thruster assembly was illuminated from the side, producing the most severe temperatures and thermal gradients across the thruster structure. A 500 hour test was performed in which thruster performance was assessed using electrical operating parameters, thrust balance and beam probe array measurements taken with the thruster operating at both nominal and simulated Mercury thermal conditions. The latter was produced using a metal collar equipped with electrical heaters, placed in close proximity to the thruster structure. The heater collar could be positioned and withdrawn without the need to vent the chamber and

  10. Translocation Renal Cell Carcinoma t(6;11)(p21;q12) and Sickle Cell Anemia: First Report and Review of the Literature.

    PubMed

    Chaste, Damien; Vian, Emmanuel; Verhoest, Gregory; Blanchet, Pascal

    2014-02-01

    Translocation renal cell carcinoma (RCC) is a family of rare tumors recently identified in the pediatric and young adult population. We report the first case of a young woman from French West Indies with sickle cell anemia who developed a translocation RCC t(6;11)(p21;q12). Usually people with the sickle cell condition are known to develop renal medullary carcinoma (RMC). To our knowledge, this is the first case described in the literature of a translocation RCC associated with sickle cell disease. Here we discuss the relation between translocation RCC, RMC, and sickle cell disease.

  11. Application of high-speed counter-current chromatography for isolation of triterpenes from Schisandra Chinensis (Turcz.) Baill and induction apoptosis mechanism of HSC-T6.

    PubMed

    Li, Bin; Meng, Xianjun; Zhu, Lijie; Jiao, Xinyao; Zhang, Jiachen

    2014-01-01

    Triterpenes have shown many beneficial activities in researches, but their separation and preparation usually require multiple methods. Following an initial cleaning-up step on the AB-8 macroporous resin, a preparative high-speed counter-current chromatography (HSCCC) with a two-phase solvent system comprising chloroform-n-butyl alcohol-methanol-water (10:1:7:4, v/v/v/v) was used to isolate and separate triterpenes from caculis of Schisandra Chinensis (Turcz.) baill. A total of 89 mg corosolic acid with purities of 98.5% were obtained from 400 mg crude extract in one-step elution and less than 4 h, and the structure identification was performed by UV, IR, MS, 1H NMR and 13C NMR. The inhibition on liver fibrosis activities of the triterpenes against HSC-T6 in vitro were studied by cell culture methods. The results showed that the corosolic acid have better inhibitory effects on HSC-T6 cells with the IC50 value of 5~25 μg/mL and the study also indicated that corosolic acid might be a potential Chinese medical component to inhibit liver fibrosis.

  12. RUNX1 truncation resulting from a cryptic and novel t(6;21)(q25;q22) chromosome translocation in acute myeloid leukemia: A case report

    PubMed Central

    Panagopoulos, Ioannis; Torkildsen, Synne; Gorunova, Ludmila; Ulvmoen, Aina; Tierens, Anne; Zeller, Bernward; Heim, Sverre

    2016-01-01

    Fluorescence in situ hybridization examination of a pediatric AML patient whose bone marrow cells carried trisomy 4 and FLT3-ITD mutation, demonstrated that part of the RUNX1 probe had unexpectedly moved to chromosome band 6q25 indicating a cryptic t(6;21)(q25;q22) translocation. RNA sequencing showed fusion of exon 7 of RUNX1 with an intergenic sequence of 6q25 close to the MIR1202 locus, something that was verified by RT-PCR together with Sanger sequencing. The RUNX1 fusion transcript encodes a truncated protein containing the Runt homology domain responsible for both heterodimerization with CBFB and DNA binding, but lacking the proline-, serine-, and threonine-rich (PST) region which is the transcription activation domain at the C terminal end. Which genetic event (+4, FLT3-ITD, t(6;21)-RUNX1 truncation or other, undetected acquired changes) was more pathogenetically important in the present case of AML, remains unknown. The case illustrates that submicroscopic chromosomal rearrangements may accompany visible numerical changes and perhaps should be actively looked for whenever a single trisomy is found. An active search for them may provide both pathogenetic and prognostic novel information. PMID:27667292

  13. A translocation t(6;14) in two cases of leiomyosarcoma: Molecular cytogenetic and array-based comparative genomic hybridization characterization.

    PubMed

    de Graaff, Marieke A; de Jong, Daniëlle; Briaire-de Bruijn, Inge H; Hogendoorn, Pancras C W; Bovée, Judith V M G; Szuhai, Károly

    2015-11-01

    Leiomyosarcomas are malignant mesenchymal tumors that recapitulate smooth muscle cell differentiation. Tumors are characterized by a genetic heterogeneity with complex karyotypes without a tumor-specific genetic aberration. Their pathobiology is still poorly understood and no specific targeted treatment is currently available for these aggressive tumors. For six leiomyosarcomas, cells were cultured and analyzed by combined binary ratio labeling fluorescence in situ hybridization (COBRA-FISH) karyotyping. A t(6;14) was identified in two cases. FISH breakpoint mapping of case L1339 reveals a breakpoint at chromosome 6p21.31 close to HMGA1, and a small deletion was observed on the distal side of the gene. A small homozygous deletion was also found in the breakpoint region of chromosome 14q24.1 involving ACTN1. The second case revealed a der(6)t(6;14)(p21.1;q21.3), with a duplication adjacent to the breakpoint at chromosome 6. Confirmatory FISH revealed a second leiomyosarcoma with an aberration at 14q24.1. Alterations at this locus were found in 5% (2 of 39) of the leiomyosarcomas in this study. The other identified breakpoints appeared to be non-recurrent, because they were not detected in other leiomyosarcomas, uterine leiomyomas, undifferentiated spindle cell sarcomas, or undifferentiated pleomorphic sarcomas.

  14. Quantum chemical study of the enzymatic repair of T(6-4)C/C(6-4)T UV-photolesions by DNA photolyases.

    PubMed

    Faraji, Shirin; Wirz, Lukas; Dreuw, Andreas

    2013-08-26

    Several strategies have evolved to repair one of the abundant UV radiation-induced damages caused to DNA, namely the mutagenic pyrimidine (6-4) pyrimidone photolesions. DNA (6-4)-photolyases are enzymes repairing these lesions by a photoinitiated electron transfer. An important aspect of a possible repair mechanism is its generality and transferability to different (6-4) lesions. Therefore, previously suggested mechanisms for the repair of the T(6-4)T lesion are here transferred to the T(6-4)C and C(6-4)T lesions and investigated theoretically using quantum chemical methods. Despite the different functional groups of the pyrimidine bases involved, a general valid molecular mechanism was identified, in which the initial step is an electron transfer coupled to a proton transfer from the protonated HIS365 to the N3(') nitrogen of the 3(') pyrimidine, followed by an intramolecular OH/NH2 transfer in one concerted step, which does not require an oxetane/azetidine or isolated water/ammonia intermediate.

  15. Time exposure studies on stress corrosion cracking of aluminum 2014-T6, aluminum 7075-T651, and titanium 6Al-4V

    NASA Technical Reports Server (NTRS)

    Terrell, J.

    1972-01-01

    The effect of a constant applied stress in crack initiation of aluminum 2014-T6, 7075-T651 and titanium 6A1-4V has been investigated. Aluminum c-ring specimens (1-inch diameter) and u-band titanium samples were exposed continuously to a 3.5% NaCl solution (pH 6) and organic fluids of ethyl, methyl, and iso-propyl alcohol (reagent purity). Corrosive action was observed to begin during the first and second day of constant exposure as evidenced by accumulation of hydrogen bubbles on the surface of stressed aluminum samples. However, a similar observation was not noted for titanium stressed specimens. Results of this investigation seems to suggest that aluminum 2014-T6, aluminum 7075-T651 are susceptible to stress corrosion cracking in chloride solution (NaCl); while they (both alloys) seem to resist stress corrosion cracking in methyl alcohol, ethyl alcohol, iso-propyl alcohol, and demineralized distilled water. Titanium 6A1-4V showed some evidence of susceptibility to SCC in methanol, while no such susceptibility was exhibited in ethanol, iso-propyl alcohol and demineralized distilled water.

  16. Radio Flaring from the T6 Dwarf WISEPC J112254.73+255021.5 with a Possible Ultra-short Periodicity

    NASA Astrophysics Data System (ADS)

    Route, Matthew; Wolszczan, Alexander

    2016-04-01

    We present new results from a continuing 5 GHz search for flaring radio emission from a sample of L and T brown dwarfs, conducted with the 305 m Arecibo radio telescope. In addition to the previously reported flaring from the T6.5 dwarf 2MASS J10475385+212423, we have detected and confirmed circularly polarized flares from another T6 dwarf, WISEPC J112254.73+255021.5. Although the flares are sporadic, they appear to occur at a stable period of 0.288 hr. Given the current constraints, periods equal to its second and third subharmonic cannot be ruled out. The stability of this period over the eight-month timespan of observations indicates that, if real, it likely reflects the star’s rapid rotation. If confirmed, any of the three inferred periodicities would be much shorter than the shortest, 1.41 hr, rotation period of a brown dwarf measured so far. This finding would place a new observational constraint on the angular momentum evolution and rotational stability of substellar objects. The detection of radio emission from the sixth ˜1000 K dwarf further demonstrates that the coolest brown dwarfs and, possibly, young giant planets, can be efficiently investigated using radio observations at centimeter wavelengths as a tool.

  17. Cryptolepine derivative-6h inhibits liver fibrosis in TGF-β1-induced HSC-T6 cells by targeting the Shh pathway.

    PubMed

    He, Ying-Hua; Li, Zeng; Ni, Ming-Ming; Zhang, Xing-Yan; Li, Ming-Fang; Meng, Xiao-Ming; Huang, Cheng; Li, Jun

    2016-09-01

    Liver fibrosis is a worldwide problem with a significant morbidity and mortality. Cryptolepis sanguinolenta (family Periplocaceae) is widely used in West African countries for the treatment of malaria, as well as for some other diseases. However, the role of C. sanguinolenta in hepatic fibrosis is still unknown. It has been reported that Methyl-CpG binding protein 2 (MeCP2) had a high expression in liver fibrosis and played a central role in its pathobiology. Interestingly, we found that a cryptolepine derivative (HZ-6h) could inhibit liver fibrosis by reducing MeCP2 expression, as evidenced by the dramatic downregulation of α-smooth muscle actin (α-SMA) and type I collagen alpha-1 (Col1α1) in protein levels in vitro. Meanwhile, we also found that HZ-6h could reduce the cell viability and promote apoptosis of hepatic stellate cells (HSCs) treated with transforming growth factor beta 1(TGF-β1). Then, we investigated the potential molecular mechanisms and found that HZ-6h blocked Shh signaling in HSC-T6 cells, resulting in the decreased protein expression of Patched-1 (PTCH-1), Sonic hedgehog (Shh), and glioma-associated oncogene homolog 1 (GLI1). In short, these results indicate that HZ-6h inhibits liver fibrosis by downregulating MeCP2 through the Shh pathway in TGF-β1-induced HSC-T6 cells. PMID:27295431

  18. Effects of Cyclic Loading on Mechanical Behavior of 24S-T4 and 75S-T6 Aluminum Alloys and SAE 4130 Steel

    NASA Technical Reports Server (NTRS)

    Macgregor, C W; Grossman, N

    1952-01-01

    An investigation was conducted to determine the effects of cyclic loading on the mechanical behavior of 24S-T4 and 75S-T6 aluminum alloys and SAE 4130 steel. Specimens of the three materials were subjected to various numbers of prior fatigue cycles both below and above the fatigue limits. Special slow-bend tests were employed to show the effects of prior cycles of fatigue stressing on the transition temperature to brittle fracture for SAE 4130 steel and on the energy-absorption capacity of the aluminum alloys. Micrographic studies were made to observe and measure crack formation and propagation and additional special tests were conducted to supplement the results of the slow-bend tests. These included Charpy impact tests, mirohardness surveys, tension tests, and fretting-corrosion studies.

  19. Study of mechanical joint strength of aluminum alloy 7075-T6 and dual phase steel 980 welded by friction bit joining and weld-bonding under corrosion medium

    SciTech Connect

    Lim, Yong Chae; Squires, Lile; Pan, Tsung-Yu; Miles, Michael; Song, Guang-Ling; Wang, Yanli; Feng, Zhili

    2014-12-30

    We have employed a unique solid-sate joining process, called friction bit joining (FBJ), to spot weld aluminum alloy (AA) 7075-T6 and dual phase (DP) 980 steel. Static joint strength was studied in the lap shear tension configuration. In addition, weld-bonding (adhesive + FBJ) joints were studied in order to evaluate the ability of adhesive to mitigate the impact of corrosion on joint properties. Accelerated laboratory cyclic corrosion tests were carried out for both FBJ only and weld-bonding joints. Furthermore, the FBJ only joints that emerged from corrosion testing had lap shear failure loads that were significantly lower than freshly prepared joints. However, weld-bonding specimens retained more than 80% of the lap shear failure load of the freshly prepared weld-bonding specimens. Moreover, examination of joint cross sections confirmed that the presence of adhesive in the weld-bonding joints mitigated the effect of the corrosion environment, compared to FBJ only joints.

  20. Influences of post weld heat treatment on tensile strength and microstructure characteristics of friction stir welded butt joints of AA2014-T6 aluminum alloy

    NASA Astrophysics Data System (ADS)

    Rajendran, C.; Srinivasan, K.; Balasubramanian, V.; Balaji, H.; Selvaraj, P.

    2016-08-01

    Friction stir welded (FSWed) joints of aluminum alloys exhibited a hardness drop in both the advancing side (AS) and retreating side (RS) of the thermo-mechanically affected zone (TMAZ) due to the thermal cycle involved in the FSW process. In this investigation, an attempt has been made to overcome this problem by post weld heat treatment (PWHT) methods. FSW butt (FSWB) joints of Al-Cu (AA2014-T6) alloy were PWHT by two methods such as simple artificial aging (AA) and solution treatment followed by artificial aging (STA). Of these two treatments, STA was found to be more beneficial than the simple aging treatment to improve the tensile properties of the FSW joints of AA2014 aluminum alloy.

  1. Investigation of Microstructure and Microhardness in Self-Reacting Friction Stir Welded AA2014-T6 and AA2219-T87

    NASA Technical Reports Server (NTRS)

    Horton, K. Renee; McGill, Preston; Barkey, Mark

    2011-01-01

    Friction stir welding (FSW) is a solid state welding process with potential advantages for aerospace and automotive industries dealing with light alloys. Self-reacting friction stir welding (SR-FSW) is one variation of the FSW process being developed at the National Aeronautics and Space Administration (NASA) for use in the fabrication of propellant tanks. This work reports on the microstructure and microhardness of SR-FSW between two dissimilar aluminum alloys. Specifically, the study examines the cross section of the weld joint formed between an AA2014-T6 plate on the advancing side and an AA2219-T87 plate on the retreating side. The microstructural analysis shows an irregularly displaced weld seam from the advancing side past the thermo-mechanical affected zone (TMAZ) into the weld nugget region. There are sharp variations in the microhardness across the weld. These variations are described in the paper and mechanisms for their formation are discussed.

  2. The young's modulus of 1018 steel and 67061-T6 aluminum measured from quasi-static to elastic precursor strain-rates

    SciTech Connect

    Rae, Philip J; Trujillo, Carl; Lovato, Manuel

    2009-01-01

    The assumption that Young's modulus is strain-rate invariant is tested for 6061-T6 aluminium alloy and 1018 steel over 10 decades of strain-rate. For the same billets of material, 3 quasi-static strain-rates are investigated with foil strain gauges at room temperature. The ultrasonic sound speeds are measured and used to calculate the moduli at approximately 10{sup 4} s{sup -1}. Finally, ID plate impact is used to generate an elastic pre-cursor in the alloys at a strain-rate of approximately 10{sup 6} s{sup -1} from which the longitudinal sound speed may be obtained. It is found that indeed the Young's modulus is strain-rate independent within the experimental accuracy.

  3. Effects of aging treatment and heat input on the microstructures and mechanical properties of TIG-welded 6061-T6 alloy joints

    NASA Astrophysics Data System (ADS)

    Peng, Dong; Shen, Jun; Tang, Qin; Wu, Cui-ping; Zhou, Yan-bing

    2013-03-01

    Aging treatment and various heat input conditions were adopted to investigate the microstructural evolution and mechanical properties of TIG welded 6061-T6 alloy joints by microstructural observations, microhardness tests, and tensile tests. With an increase in heat input, the width of the heat-affected zone (HAZ) increases and grains in the fusion zone (FZ) coarsen. Moreover, the hardness of the HAZ decreases, whereas that of the FZ decreases initially and then increases with an increase in heat input. Low heat input results in the low ultimate tensile strength of the welded joints due to the presence of partial penetrations and pores in the welded joints. After a simple artificial aging treatment at 175°C for 8 h, the microstructure of the welded joints changes slightly. The mechanical properties of the welded joints enhance significantly after the aging process as few precipitates distribute in the welded seam.

  4. Paternal origin of der(X)t(X;6) in a girl with trisomy 6p and unbalanced t(6;10) mosaicism in her mother.

    PubMed

    Petković, Iskra; Barisić, Ingeborg; Bastić, Mislav; Hećimović, Silva; Bago, Ruzica

    2003-07-15

    We present a case of trisomy for the whole short arm of chromosome 6 in a 3-year-old girl with moderate mental retardation, mild facial dysmorphism, short stature, failure to thrive, and no abnormalities of the visceral organs. Cytogenetic and fluorescence in situ hybridization (FISH) analysis revealed a 46, X, der(X)t(X;6)(q22; p11.1) karyotype. The derived X was late replicating with variable spreading of X chromosome inactivation onto the translocated 6p. A normal karyotype was observed in the father, while the mother presented 46,XX/46,XX, der(10)t(6;10)(p11;p11). The mother is a mosaic with unbalanced t(6;10) in 4.7% of cells. To the best of our knowledge, this unusual mosaicism has not yet been reported. In this family the short arm of chromosome 6 was involved in an unbalanced rearrangement with chromosome X in the proband and with chromosome 10 in the mother. In order to study the mechanism of the formation of t(X;6) in the girl we performed DNA polymorphism analysis. These investigations revealed that chromosomes X and 6 involved in the rearrangement are of paternal origin. Our patient exhibits only discrete facial features characteristic of partial trisomy 6p. We suggest that mild phenotypic expression be probably due to X chromosome inactivation spreading onto the translocated 6p. This report show that combined cytogenetic, FISH, and molecular analysis of chromosomal aberrations are necessary for the understanding of the mechanism of formation, parental origin, and genetic counseling. PMID:12833412

  5. Molecular confirmation of t(6;11)(p21;q12) renal cell carcinoma in archival paraffin-embedded material using a break-apart TFEB FISH assay expands its clinicopathologic spectrum.

    PubMed

    Argani, Pedram; Yonescu, Raluca; Morsberger, Laura; Morris, Kerry; Netto, George J; Smith, Nathan; Gonzalez, Nilda; Illei, Peter B; Ladanyi, Marc; Griffin, Constance A

    2012-10-01

    A subset of renal cell carcinomas (RCCs) is characterized by t(6;11)(p21;q12), which results in fusion of the untranslated Alpha (MALAT1) gene to the TFEB gene. Only 21 genetically confirmed cases of t(6;11) RCCs have been reported. This neoplasm typically demonstrates a distinctive biphasic morphology, comprising larger epithelioid cells and smaller cells clustered around basement membrane material; however, the full spectrum of its morphologic appearances is not known. The t(6;11) RCCs differ from most conventional RCCs in that they consistently express melanocytic immunohistochemical (IHC) markers such as HMB45 and Melan A and the cysteine protease cathepsin K but are often negative for epithelial markers such as cytokeratins. TFEB IHC has been proven to be useful to confirm the diagnosis of t(6;11) RCCs in archival material, because native TFEB is upregulated through promoter substitution by the gene fusion. However, IHC is highly fixation dependent and has been proven to be particularly difficult for TFEB. A validated fluorescence in situ hybridization (FISH) assay for molecular confirmation of the t(6;11) RCC in archival formalin-fixed, paraffin-embedded material has not been previously reported. We report herein the development of a break-apart TFEB FISH assay for the diagnosis of t(6;11)(p21;q12) RCCs. We validated the assay on 4 genetically confirmed cases and 76 relevant expected negative control cases and used the assay to report 8 new cases that expand the clinicopathologic spectrum of t(6;11) RCCs. An additional previously reported TFEB IHC-positive case was confirmed by TFEB FISH in 46-year-old archival material. In conclusion, TFEB FISH is a robust, clinically validated assay that can confirm the diagnosis of t(6;11) RCC in archival material and should allow a more comprehensive clinicopathologic delineation of this recently recognized neoplastic entity.

  6. Critical assessment of precracked specimen configuration and experimental test variables for stress corrosion testing of 7075-T6 aluminum alloy plate

    NASA Technical Reports Server (NTRS)

    Domack, M. S.

    1985-01-01

    A research program was conducted to critically assess the effects of precracked specimen configuration, stress intensity solutions, compliance relationships and other experimental test variables for stress corrosion testing of 7075-T6 aluminum alloy plate. Modified compact and double beam wedge-loaded specimens were tested and analyzed to determine the threshold stress intensity factor and stress corrosion crack growth rate. Stress intensity solutions and experimentally determined compliance relationships were developed and compared with other solutions available in the literature. Crack growth data suggests that more effective crack length measurement techniques are necessary to better characterize stress corrosion crack growth. Final load determined by specimen reloading and by compliance did not correlate well, and was considered a major source of interlaboratory variability. Test duration must be determined systematically, accounting for crack length measurement resolution, time for crack arrest, and experimental interferences. This work was conducted as part of a round robin program sponsored by ASTM committees G1.06 and E24.04 to develop a standard test method for stress corrosion testing using precracked specimens.

  7. Study of mechanical joint strength of aluminum alloy 7075-T6 and dual phase steel 980 welded by friction bit joining and weld-bonding under corrosion medium

    DOE PAGES

    Lim, Yong Chae; Squires, Lile; Pan, Tsung-Yu; Miles, Michael; Song, Guang-Ling; Wang, Yanli; Feng, Zhili

    2014-12-30

    We have employed a unique solid-sate joining process, called friction bit joining (FBJ), to spot weld aluminum alloy (AA) 7075-T6 and dual phase (DP) 980 steel. Static joint strength was studied in the lap shear tension configuration. In addition, weld-bonding (adhesive + FBJ) joints were studied in order to evaluate the ability of adhesive to mitigate the impact of corrosion on joint properties. Accelerated laboratory cyclic corrosion tests were carried out for both FBJ only and weld-bonding joints. Furthermore, the FBJ only joints that emerged from corrosion testing had lap shear failure loads that were significantly lower than freshly preparedmore » joints. However, weld-bonding specimens retained more than 80% of the lap shear failure load of the freshly prepared weld-bonding specimens. Moreover, examination of joint cross sections confirmed that the presence of adhesive in the weld-bonding joints mitigated the effect of the corrosion environment, compared to FBJ only joints.« less

  8. Microstructure and mechanical properties of twin-wire arc sprayed Ni-Al composite coatings on 6061-T6 aluminum alloy sheet

    NASA Astrophysics Data System (ADS)

    Wang, Ji-xiao; Liu, Jing-shun; Zhang, Lun-yong; Sun, Jian-fei; Wang, Zhi-ping

    2014-05-01

    We have systematically studied the microstructure and mechanical properties of Ni-5wt%Al and Ni-20wt%Al composite coatings fabricated on 6061-T6 aluminum alloy sheet by twin-wire arc spraying under different experimental conditions. The abrasive wear behavior and interface diffusion behavior of the composite coatings were evaluated by dry/wet rubber wheel abrasive wear tests and heat treatment, respectively. Experimental results indicate that the composite coatings exhibit features of adhesive wear. Besides, the Vickers microhardness of NiAl and Ni3Al intermetallic compounds is relatively larger than that of the substrate, which is beneficial for enhancing the wear resistance. With the increase of annealing temperature and time, the interface diffusion area between the Ni-Al coating and the substrate gradually expands with the formation of NiAl3 and Ni2Al3 phases, and is controlled by diffusion of aluminum atoms. The grain growth exponent n of diffusion kinetics of the Ni-Al coating, calculated via a high-temperature diffusion model at 400, 480, and 550°C, is between 0.28 and 0.38. This satisfies the cubic law, which is consistent with the general theoretical relationship of high-temperature diffusion.

  9. Effect of Welding Parameters on Microstructure, Thermal, and Mechanical Properties of Friction-Stir Welded Joints of AA7075-T6 Aluminum Alloy

    NASA Astrophysics Data System (ADS)

    Lotfi, Amir Hossein; Nourouzi, Salman

    2014-06-01

    A high-strength Al-Zn-Mg-Cu alloy AA7075-T6 was friction-stir welded with various process parameter combinations incorporating the design of the experiment to investigate the effect of welding parameters on the microstructure and mechanical properties. A three-factors, five-level central composition design (CCD) has been used to minimize the number of experimental conditions. The friction-stir welding parameters have significant influence on the heat input and temperature profile, which in turn regulates the microstructural and mechanical properties of the joints. The weld thermal cycles and transverse distribution of microhardness of the weld joints were measured, and the tensile properties were tested. The fracture surfaces of tensile specimens were observed by a scanning electron microscope (SEM), and the formation of friction-stir processing zone has been analyzed macroscopically. Also, an equation was derived to predict the final microhardness and tensile properties of the joints, and statistical tools are used to develop the relationships. The results show that the peak temperature during welding of all the joints was up to 713 K (440 °C), which indicates the key role of the tool shoulder diameter in deciding the maximum temperature. From this investigation, it was found that the joint fabricated at a rotational speed of 1050 rpm, welding speed of 100 mm/min, and shoulder diameter of 14 mm exhibited higher mechanical properties compared to the other fabricated joints.

  10. Influence of MC3T3-E1 preosteoblast culture on the corrosion of a T6-treated AZ91 alloy

    PubMed Central

    Brooks, Emily K.; Tobias, Menachem E.; Yang, Shuying; Bone, Lawrence B.; Ehrensberger, Mark T.

    2015-01-01

    This study investigated the corrosion of artificially aged T6 heat-treated Mg-9%Al-1%Zn (AZ91) for biomedical applications. Corrosion tests and surface analysis were completed both with and without a monolayer of mouse preosteoblast MC3T3-E1 cells cultured on the sample. Electrochemical impedance spectroscopy (EIS) and inductively coupled plasma mass spectroscopy (ICPMS) were used to explore the corrosion processes after either 3 or 21 days of AZ91 incubation in cell culture medium (CCM). The EIS showed both the inner layer resistance (Rin) and outer layer resistance (Rout) were lower for samples without cells cultured on the surface at 3 days (Rin = 2.64 e4 Ω/cm2, Rout = 140 Ω/cm2) compared to 21 days (Rin = 3.60 e4 Ω/cm2, Rout = 287 Ω/cm2) due to precipitation of magnesium and calcium phosphates over time. Samples with preosteoblasts cultured on the surface had a slower initial corrosion (3 day, Rin = 1.88 e5 Ω/cm2, Rout = 1060 Ω/cm2) which was observed to increase over time (21 day, Rin = 2.99 e4 Ω/cm2, Rout = 287 Ω/cm2). Changes in the corrosion processes were thought to be related to changes in the coverage provided by the cell layer. Our results reveal that the presence of cells and biological processes are able to significantly influence the corrosion rate of AZ91. PMID:25715925

  11. Effect of Porosity on the Fatigue Life of Cast AC4C-T6 Alloy for Lcd Glass Transfer Robot

    NASA Astrophysics Data System (ADS)

    Shim, Hee-Jin; Kim, Jung-Kyu

    Aluminium has good corrosion properties and a high strength to weight reduction which makes it favourable in many applications. The increased use of aluminium casting in the automotive industry does also imply that the need for design data for aluminium increases. Especially for castings, the influences of casting defects are always an issue. For this reason fatigue properties for as-cast sand and permanent mould specimens with different contents of porosity have been studied. The cast aluminium specimens of two different porosities were fatigue tested in cyclic axial test at R=-1. Prior to fatigue test specimens were examined by CT-scan and sorted into two quality groups depending on the porosity level. The aim of this work was to investigate the fatigue life for cast AC4C-T6 alloy with different amounts of inherent porosity. An additional aim was to predict the durability for cast components with defect constrained in a specified volume of components, by using a commercial program MSC. Fatigue.

  12. Design, fabrication and test of a hydrogen heat pipe. [extruding and grooving 6063-T6 aluminum tubes for cryogenic heat pipes

    NASA Technical Reports Server (NTRS)

    Alario, J.

    1979-01-01

    Re-entrant groove technology was extended to hydrogen heat pipes. Parametric analyses are presented which optimize the theoretical design while considering the limitations of state-of-the-art extrusion technology. The 6063-T6 aluminum extrusion is 14.6 mm OD with a wall thickness of 1.66 mm and contains 20 axial grooves which surround a central 9.3 mm diameter vapor core. Each axial groove is 0.775 mm diameter with a 0.33 mm opening. An excess vapor reservoir is provided at the evaporator to minimize the pressure containment hazard during ambient storage. Modifications to the basic re-entrant groove profile resulted in improved overall performance. While the maximum heat transport capacity decreased slightly to 103 w-m the static wicking height increased markedly to 4.5 cm. The heat pipe became operational between 20 and 30 K after a cooldown from 77 K without any difficulty. Steady state performance data taken over a 19 to 23 K temperature range indicated: (1) maximum heat transport capacity of 5.4 w-m; (2) static wicking height of 1.42 cm; and (3) overall heat pipe conductance of 1.7 watts/deg C.

  13. Severe mental retardation in six generations of a large South African family carrying a translocation t(6;10)(q27;q25.2).

    PubMed Central

    Brusnický, J; van Heerden, K M; de Jong, G; Cronjé, A S; Retief, A E

    1986-01-01

    Partial monosomy 10q25.2----qter, detected in a newborn baby with multiple congenital abnormalities, was found to be derived from a balanced maternal translocation t(6;10)(q27;q25.2). The pedigree of six generations of the family is presented. In an extensive cytogenetic study of this family, the chromosome complements of 57 subjects, potentially capable of carrying some form of this translocation, were analysed. A total of 14 male carriers (four obligatory) and 14 female carriers (three obligatory) of this translocation was found. Partial trisomy 10q25.2----qter, associated with severe mental retardation, occurred in nine cases, eight males and one female. Two of these eight males were detected prenatally and subsequently therapeutically aborted. The phenotypes of the family members with partial trisomy 10q25.2----qter are compared to each other and to those reported in publications. No further cases of partial monosomy 10q25.2----qter were encountered. A review of published reports of partial monosomy and partial trisomy 10qter is given. The apparent absence of infertility, the occurrence of many first trimester miscarriages, and the marked sex ratio are discussed. Images PMID:3783620

  14. Molecular genetic analysis of a de novo balanced translocation t(6;17)(p21.31;q11.2) associated with hypospadias and anorectal malformation.

    PubMed

    Mansouri, Mahmoud Reza; Carlsson, Birgit; Davey, Edward; Nordenskjöld, Agneta; Wester, Tomas; Annerén, Göran; Läckgren, Göran; Dahl, Niklas

    2006-03-01

    We report a young boy with penoscrotal hypospadias, anal atresia (AA) with a recto-urethral fistula, a hypoplastic kidney and a balanced translocation t(6;17)(p21.31;q11.2). Physical mapping of the breakpoints localized the chromosome 6 breakpoint within an intron of the gene lipoma HMGIC fusion partner-like 5 (LHFPL5) whereas the chromosome 17 breakpoint was mapped to the first intron of the 182-FIP gene encoding the Fragile X Mental Retardation Protein Interacting Protein. Sequence analysis across the breakpoints revealed an almost perfectly balanced translocation with a 2 bp deletion on the derivative chromosome 6 and a 7 bp duplication on the derivative chromosome 17. We identified a fusion transcript consisting of the first exon of 182-FIP and the last exon of LHFPL5 in patient-derived cells. Quantitative expression analysis of the genes flanking the breakpoints, revealed increased transcript levels for SFRS protein kinase 1 (SRPK1) and TAO kinase 1 (TAOK1) which suggests a positional effect due to the translocation. We hypothesize that the urogenital and anorectal malformations in the patient result from one or several mechanisms including disruption of the genes 182-FIP and LHFPL5, altered expression of the genes flanking the translocation breakpoints and, a gain of function mechanism mediated by the 182-FIP-LHFPL5 fusion transcript. PMID:16395596

  15. Coating formation by plasma electrolytic oxidation on ZC71/SiC/12p-T6 magnesium metal matrix composite

    NASA Astrophysics Data System (ADS)

    Arrabal, R.; Matykina, E.; Skeldon, P.; Thompson, G. E.

    2009-02-01

    Plasma electrolytic oxidation (PEO) of a ZC71/SiC/12p-T6 magnesium metal matrix composite (MMC) is investigated in relation to coating growth and corrosion behaviour. PEO treatment was undertaken at 350 mA cm -2 (rms) and 50 Hz with a square waveform in stirred 0.05 M Na 2SiO 3.5H 2O/0.1 M KOH electrolyte. The findings revealed thick, dense oxide coatings, with an average hardness of 3.4 GPa, formed at an average rate of ˜1 μm min -1 for treatment times up to 100 min and ˜0.2 μm min -1 for later times. The coatings are composed mainly of MgO and Mg 2SiO 4, with an increased silicon content in the outer regions, constituting <10% of the coating thickness. SiC particles are incorporated into the coating, with formation of a silicon-rich layer at the particle/coating interface due to exposure to high temperatures during coating formation. The distribution of the particles in the coating indicated growth of new oxide at the metal/coating interface. The corrosion rate of the MMC in 3.5% NaCl is reduced by approximately two orders of magnitude by the PEO treatment.

  16. Renal cell carcinomas with t(6;11)(p21;q12) presenting with tubulocystic renal cell carcinoma-like features.

    PubMed

    Rao, Qiu; Zhang, Xiu-Mei; Tu, Pin; Xia, Qiu-Yuan; Shen, Qin; Zhou, Xiao-Jun; Shi, Qun-Li

    2013-01-01

    In this study, we reported an additional genetically confirmed case of renal cell carcinomas (RCCs) with t(6;11)(p21;q12) showing an unusual histological pattern. Histologically, the tumor was entirely composed of small to intermediate sized tubules and cysts. The tubules and cysts were lined by a single layer of flat, hobnail, cuboidal to columnar epithelial cells. Most cells demonstrated abundant eosinophilic cytoplasm with regular, round or oval nuclei and some inconspicuous nucleoli. All these morphological features are suggestive of tubulocystic carcinoma of the kidney. However, the tumor demonstrated moderately (2+) or strongly (3+) positive staining for TFEB, Cathepsin K, Ksp-cadherin, and vimentin but negative for TFE3, CD10, HMB45, melan A, CKpan, and CK7. Using a recently developed TFEB split FISH assay, the presence of TFEB rearrangement was demonstrated. Our results support the clinical application of a TFEB break-apart FISH assay for diagnosis and confirmation of TFEB RCC and further expand the morphologic spectrum that may be present in these neoplasms, sometimes raising a challenging differential diagnosis with other renal tumors.

  17. Renal cell carcinomas with t(6;11)(p21;q12) presenting with tubulocystic renal cell carcinoma-like features.

    PubMed

    Rao, Qiu; Zhang, Xiu-Mei; Tu, Pin; Xia, Qiu-Yuan; Shen, Qin; Zhou, Xiao-Jun; Shi, Qun-Li

    2013-01-01

    In this study, we reported an additional genetically confirmed case of renal cell carcinomas (RCCs) with t(6;11)(p21;q12) showing an unusual histological pattern. Histologically, the tumor was entirely composed of small to intermediate sized tubules and cysts. The tubules and cysts were lined by a single layer of flat, hobnail, cuboidal to columnar epithelial cells. Most cells demonstrated abundant eosinophilic cytoplasm with regular, round or oval nuclei and some inconspicuous nucleoli. All these morphological features are suggestive of tubulocystic carcinoma of the kidney. However, the tumor demonstrated moderately (2+) or strongly (3+) positive staining for TFEB, Cathepsin K, Ksp-cadherin, and vimentin but negative for TFE3, CD10, HMB45, melan A, CKpan, and CK7. Using a recently developed TFEB split FISH assay, the presence of TFEB rearrangement was demonstrated. Our results support the clinical application of a TFEB break-apart FISH assay for diagnosis and confirmation of TFEB RCC and further expand the morphologic spectrum that may be present in these neoplasms, sometimes raising a challenging differential diagnosis with other renal tumors. PMID:23826432

  18. Influence of MC3T3-E1 preosteoblast culture on the corrosion of a T6-treated AZ91 alloy.

    PubMed

    Brooks, Emily K; Tobias, Menachem E; Yang, Shuying; Bone, Lawrence B; Ehrensberger, Mark T

    2016-02-01

    This study investigated the corrosion of artificially aged T6 heat-treated Mg-9%Al-1%Zn (AZ91) for biomedical applications. Corrosion tests and surface analysis were completed both with and without a monolayer of mouse preosteoblast MC3T3-E1 cells cultured on the sample. Electrochemical impedance spectroscopy (EIS) and inductively coupled plasma mass spectroscopy (ICPMS) were used to explore the corrosion processes after either 3 or 21 days of AZ91 incubation in cell culture medium (CCM). The EIS showed both the inner layer resistance (Rin ) and outer layer resistance (Rout ) were lower for samples without cells cultured on the surface at 3 days (Rin  = 2.64 e4 Ω/cm(2) , Rout  = 140 Ω/cm(2) ) compared to 21 days (Rin  = 3.60 e4 Ω/cm(2) , Rout  = 287 Ω/cm(2) ) due to precipitation of magnesium and calcium phosphates over time. Samples with preosteoblasts cultured on the surface had a slower initial corrosion (3 day, Rin  = 1.88 e5 Ω/cm(2) , Rout  = 1060 Ω/cm(2) ) which was observed to increase over time (21 day, Rin  = 2.99 e4 Ω/cm(2) , Rout  = 287 Ω/cm(2) ). Changes in the corrosion processes were thought to be related to changes in the coverage provided by the cell layer. Our results reveal that the presence of cells and biological processes are able to significantly influence the corrosion rate of AZ91.

  19. Protein engineering by random mutagenesis and structure-guided consensus of Geobacillus stearothermophilus Lipase T6 for enhanced stability in methanol.

    PubMed

    Dror, Adi; Shemesh, Einav; Dayan, Natali; Fishman, Ayelet

    2014-02-01

    The abilities of enzymes to catalyze reactions in nonnatural environments of organic solvents have opened new opportunities for enzyme-based industrial processes. However, the main drawback of such processes is that most enzymes have a limited stability in polar organic solvents. In this study, we employed protein engineering methods to generate a lipase for enhanced stability in methanol, which is important for biodiesel production. Two protein engineering approaches, random mutagenesis (error-prone PCR) and structure-guided consensus, were applied in parallel on an unexplored lipase gene from Geobacillus stearothermophilus T6. A high-throughput colorimetric screening assay was used to evaluate lipase activity after an incubation period in high methanol concentrations. Both protein engineering approaches were successful in producing variants with elevated half-life values in 70% methanol. The best variant of the random mutagenesis library, Q185L, exhibited 23-fold-improved stability, yet its methanolysis activity was decreased by one-half compared to the wild type. The best variant from the consensus library, H86Y/A269T, exhibited 66-fold-improved stability in methanol along with elevated thermostability (+4.3°C) and a 2-fold-higher fatty acid methyl ester yield from soybean oil. Based on in silico modeling, we suggest that the Q185L substitution facilitates a closed lid conformation that limits access for both the methanol and substrate excess into the active site. The enhanced stability of H86Y/A269T was a result of formation of new hydrogen bonds. These improved characteristics make this variant a potential biocatalyst for biodiesel production. PMID:24362426

  20. Study on the 8He ground state via 8He(p, d)7He and 8He(p, t)6He reaction at 82.3 MeV/nucleon

    NASA Astrophysics Data System (ADS)

    Lou, J. L.; Ye, Y. L.; Pang, D. Y.; Cao, Z. X.; Jiang, D. X.

    2013-03-01

    The differential cross sections for reactions 8He(p, d)7Heg.s and 8He(p, t)6He2+ were measured by the coincidence of d + 6 He and t + 4He in a wide angular range from 15 to 130 degree in the center-of-mass system. The cross sections of the 8He(p, t)6Heg.s reaction were extremely lower than that of 8He(p, t)6He2+ reaction, and was not obtained from the present data. The experimental results were compared to the preliminary theoretical calculations in the frame of adiabatic distorted-wave approximation with the code Fresco. The results show that the configuration of (1p3/2)2(1p1/2)2 may have some probability in the ground state wave function of 8He. The cluster structure of 8He = 5H + 3H could not be neglected.

  1. Microstructure and mechanical properties of 2024-T3 and 7075-T6 aluminum alloys and austenitic stainless steel 304 after being exposed to hydrogen peroxide

    NASA Astrophysics Data System (ADS)

    Sofyan, Nofrijon Bin Imam

    The effect of hydrogen peroxide used as a decontaminant agent on selected aircraft metallic materials has been investigated. The work is divided into three sections; bacterial attachment behavior onto an austenitic stainless steel 304 surface; effect of decontamination process on the microstructure and mechanical properties of aircraft metallic structural materials of two aluminum alloys, i.e. 2024-T3 and 7075-T6, and an austenitic stainless steel 304 as used in galley and lavatory surfaces; and copper dissolution rate into hydrogen peroxide. With respect to bacterial attachment, the results show that surface roughness plays a role in the attachment of bacteria onto metallic surfaces at certain extent. However, when the contact angle of the liquid on a surface increased to a certain degree, detachment of bacteria on that surface became more difficult. In its relation to the decontamination process, the results show that a corrosion site, especially on the austenitic stainless steel 304 weld and its surrounding HAZ area, needs more attention because it could become a source or a harborage of bio-contaminant agent after either incidental or intentional bio-contaminant delivery. On the effect of the decontamination process on the microstructure and mechanical properties of aircraft metallic structural materials, the results show that microstructural effects are both relatively small in magnitude and confined to a region immediately adjacent to the exposed surface. No systematic effect is found on the tensile properties of the three alloys under the conditions examined. The results of this investigation are promising with respect to the application of vapor phase hydrogen peroxide as a decontaminant agent to civilian aircraft, in that even under the most severe circumstances that could occur; only very limited damage was observed. The results from the dissolution of copper by concentrated liquid hydrogen peroxide showed that the rate of copper dissolution increased for

  2. Cloning, purification and preliminary crystallographic analysis of Ara127N, a GH127 β-L-arabinofuranosidase from Geobacillus stearothermophilus T6.

    PubMed

    Lansky, Shifra; Salama, Rachel; Dann, Roie; Shner, Izhak; Manjasetty, Babu A; Belrhali, Hassan; Shoham, Yuval; Shoham, Gil

    2014-08-01

    The L-arabinan utilization system of Geobacillus stearothermophilus T6 is composed of five transcriptional units that are clustered within a 38 kb DNA segment. One of the transcriptional units contains 11 genes, the last gene of which (araN) encodes a protein, Ara127N, that belongs to the newly established GH127 family. Ara127N shares 44% sequence identity with the recently characterized HypBA1 protein from Bifidobacterium longum and thus is likely to function similarly as a β-L-arabinofuranosidase. β-L-Arabinofuranosidases are enzymes that hydrolyze β-L-arabinofuranoside linkages, the less common form of such linkages, a unique enzymatic activity that has been identified only recently. The interest in the structure and mode of action of Ara127N therefore stems from its special catalytic activity as well as its membership of the new GH127 family, the structure and mechanism of which are only starting to be resolved. Ara127N has recently been cloned, overexpressed, purified and crystallized. Two suitable crystal forms have been obtained: one (CTP form) belongs to the monoclinic space group P21, with unit-cell parameters a = 104.0, b = 131.2, c = 107.6 Å, β = 112.0°, and the other (RB form) belongs to the orthorhombic space group P212121, with unit-cell parameters a = 65.5, b = 118.1, c = 175.0 Å. A complete X-ray diffraction data set has been collected to 2.3 Å resolution from flash-cooled crystals of the wild-type enzyme (RB form) at -173°C using synchrotron radiation. A selenomethionine derivative of Ara127N has also been prepared and crystallized for multi-wavelength anomalous diffraction (MAD) experiments. Crystals of selenomethionine Ara127N appeared to be isomorphous to those of the wild type (CTP form) and enabled the measurement of a three-wavelength MAD diffraction data set at the selenium absorption edge. These data are currently being used for detailed three-dimensional structure determination of the Ara127N protein. PMID:25084377

  3. Francisella tularensis IglG Belongs to a Novel Family of PAAR-Like T6SS Proteins and Harbors a Unique N-terminal Extension Required for Virulence

    PubMed Central

    Mosnier, Amandine; Hologne, Maggy; Martin, Amandine; Lindgren, Lena; Punginelli, Claire; Lays, Claire; Walker, Olivier; Charbit, Alain; Telouk, Philippe; Conlan, Wayne; Terradot, Laurent; Sjöstedt, Anders; Henry, Thomas

    2016-01-01

    The virulence of Francisella tularensis, the etiological agent of tularemia, relies on an atypical type VI secretion system (T6SS) encoded by a genomic island termed the Francisella Pathogenicity Island (FPI). While the importance of the FPI in F. tularensis virulence is clearly established, the precise role of most of the FPI-encoded proteins remains to be deciphered. In this study, using highly virulent F. tularensis strains and the closely related species F. novicida, IglG was characterized as a protein featuring a unique α-helical N-terminal extension and a domain of unknown function (DUF4280), present in more than 250 bacterial species. Three dimensional modeling of IglG and of the DUF4280 consensus protein sequence indicates that these proteins adopt a PAAR-like fold, suggesting they could cap the T6SS in a similar way as the recently described PAAR proteins. The newly identified PAAR-like motif is characterized by four conserved cysteine residues, also present in IglG, which may bind a metal atom. We demonstrate that IglG binds metal ions and that each individual cysteine is required for T6SS-dependent secretion of IglG and of the Hcp homologue, IglC and for the F. novicida intracellular life cycle. In contrast, the Francisella-specific N-terminal α-helical extension is not required for IglG secretion, but is critical for F. novicida virulence and for the interaction of IglG with another FPI-encoded protein, IglF. Altogether, our data suggest that IglG is a PAAR-like protein acting as a bi-modal protein that may connect the tip of the Francisella T6SS with a putative T6SS effector, IglF. PMID:27602570

  4. Francisella tularensis IglG Belongs to a Novel Family of PAAR-Like T6SS Proteins and Harbors a Unique N-terminal Extension Required for Virulence.

    PubMed

    Rigard, Mélanie; Bröms, Jeanette E; Mosnier, Amandine; Hologne, Maggy; Martin, Amandine; Lindgren, Lena; Punginelli, Claire; Lays, Claire; Walker, Olivier; Charbit, Alain; Telouk, Philippe; Conlan, Wayne; Terradot, Laurent; Sjöstedt, Anders; Henry, Thomas

    2016-09-01

    The virulence of Francisella tularensis, the etiological agent of tularemia, relies on an atypical type VI secretion system (T6SS) encoded by a genomic island termed the Francisella Pathogenicity Island (FPI). While the importance of the FPI in F. tularensis virulence is clearly established, the precise role of most of the FPI-encoded proteins remains to be deciphered. In this study, using highly virulent F. tularensis strains and the closely related species F. novicida, IglG was characterized as a protein featuring a unique α-helical N-terminal extension and a domain of unknown function (DUF4280), present in more than 250 bacterial species. Three dimensional modeling of IglG and of the DUF4280 consensus protein sequence indicates that these proteins adopt a PAAR-like fold, suggesting they could cap the T6SS in a similar way as the recently described PAAR proteins. The newly identified PAAR-like motif is characterized by four conserved cysteine residues, also present in IglG, which may bind a metal atom. We demonstrate that IglG binds metal ions and that each individual cysteine is required for T6SS-dependent secretion of IglG and of the Hcp homologue, IglC and for the F. novicida intracellular life cycle. In contrast, the Francisella-specific N-terminal α-helical extension is not required for IglG secretion, but is critical for F. novicida virulence and for the interaction of IglG with another FPI-encoded protein, IglF. Altogether, our data suggest that IglG is a PAAR-like protein acting as a bi-modal protein that may connect the tip of the Francisella T6SS with a putative T6SS effector, IglF. PMID:27602570

  5. An Experimental Investigation of the Effects of Vacuum Environment on the Fatigue Life, Fatigue-Crack-Growth Behavior, and Fracture Toughness of 7075-T6 Aluminum Alloy. Ph.D. Thesis - North Carolina State Univ.

    NASA Technical Reports Server (NTRS)

    Hudson, C. M.

    1972-01-01

    Axial load fatigue life, fatigue-crack propagation, and fracture toughness tests were conducted on 0.090-inch thick specimens made of 7075-T6 aluminum alloy. The fatigue life and fatigue-crack propagation experiments were conducted at a stress ratio of 0.02. Maximum stresses ranged from 33 to 60 ksi in the fatigue life experiments, and from 10 to 40 ksi in the fatigue-crack propagation experiments, and fatigue life experiments were conducted at gas pressures of 760, 0.5, 0.05, and 0.00000005 torr. Fatigue-crack-growth and fracture toughness experiments were conducted at gas pressures of 760 and 5 x 10 to the minus 8th power torr. Residual stress measurements were made on selected fatigue life specimens to determine the effect of such stresses on fatigue life. Analysis of the results from the fatigue life experiments indicated that fatigue life progressively increased as the gas pressure decreased. Analysis of the results from the fatigue-crack-growth experiments indicates that at low values of stress-intensity range, the fatigue crack growth rates were approximately twice as high in air as in vacuum. Fracture toughness data showed there was essentially no difference in the fracture toughness of 7075-T6 in vacuum and in air.

  6. Familial translocation t(6;20)(p21;p13) resulting in partial trisomy 6p and partial monosomy 20p: report of a new case and review of the literature.

    PubMed

    Berner, A L; Bağci, S; Wohlleber, E; Engels, E; Müller, A; Bartmann, P; Weber, R G; Reutter, H

    2012-01-01

    Carriers of completely balanced chromosomal translocations have all necessary genetic information. Nevertheless, because of the possibility of maldistribution during gametogenesis, they are at increased risk for infertility, miscarriage, stillbirth or having a child with congenital anomalies including mental retardation. As postnatal clinical reports are infrequent, prediction of clinical course for specific unbalanced karyotypes diagnosed during pregnancy remains difficult. Here, we report the 6th case of partial trisomy 6p and partial monosomy 20p due to an unbalanced adjacent-1 segregation of the rare familial translocation t(6;20)(p21;p13). We give a thorough clinical description of the present case, demonstrating broad phenotypic overlap with the 5 previously published cases reviewed here, providing important data on postnatal outcome.

  7. A 10-year wildlife survey of 15 species of Canadian carnivores identifies new hosts or geographic locations for Trichinella genotypes T2, T4, T5, and T6.

    PubMed

    Gajadhar, Alvin A; Forbes, Lorry B

    2010-02-26

    A survey of wild carnivores in Canada was conducted over a 10-year period to determine the prevalence and genotypes of Trichinella. Muscle samples collected from 1409 animals representing 15 hosts species were enzymatically digested to recover Trichinella larvae. Larvae were recovered from a total of 287 (20.4%) animals and PCR identified four genotypes of Trichinella. Trichinella nativa was found in 5 host species and was the most commonly found genotype. Trichinella T6 was present in 7 species of carnivores, and coyote and badger are new host records for this genotype. The recovery of T. pseudospiralis and T. murrelli from cougars is the first documentation of these species in Canada and in cougars. The cougar was also the only host species in which all four genotypes of Trichinella were identified. Black bears and walruses had the highest tissue levels of larvae in this study and are also the species most frequently associated with human trichinellosis in Canada. This work identifies additional host species and expanded geographic ranges for 4 genotypes of Trichinella in North America. Failure to demonstrate T. spiralis in wildlife and continued negative results from ongoing surveillance activities in swine provide additional evidence that T. spiralis is not present in Canada. PMID:19926223

  8. A 10-year wildlife survey of 15 species of Canadian carnivores identifies new hosts or geographic locations for Trichinella genotypes T2, T4, T5, and T6.

    PubMed

    Gajadhar, Alvin A; Forbes, Lorry B

    2010-02-26

    A survey of wild carnivores in Canada was conducted over a 10-year period to determine the prevalence and genotypes of Trichinella. Muscle samples collected from 1409 animals representing 15 hosts species were enzymatically digested to recover Trichinella larvae. Larvae were recovered from a total of 287 (20.4%) animals and PCR identified four genotypes of Trichinella. Trichinella nativa was found in 5 host species and was the most commonly found genotype. Trichinella T6 was present in 7 species of carnivores, and coyote and badger are new host records for this genotype. The recovery of T. pseudospiralis and T. murrelli from cougars is the first documentation of these species in Canada and in cougars. The cougar was also the only host species in which all four genotypes of Trichinella were identified. Black bears and walruses had the highest tissue levels of larvae in this study and are also the species most frequently associated with human trichinellosis in Canada. This work identifies additional host species and expanded geographic ranges for 4 genotypes of Trichinella in North America. Failure to demonstrate T. spiralis in wildlife and continued negative results from ongoing surveillance activities in swine provide additional evidence that T. spiralis is not present in Canada.

  9. A Comparative Analysis of the Impact of Tool Design to Fatigue Behavior of Single-Sided and Double-Sided Welded Butt Joints of EN AW 6082-T6 Alloy

    NASA Astrophysics Data System (ADS)

    Krasnowski, K.; Dymek, S.

    2013-12-01

    In this paper, the results of fatigue behavior on friction stir welded joints of aluminum alloy EN AW 6082-T6 are reported. In particular, the study presents the influence of the geometry of a welding tool on fatigue strengths and tensile strengths. The test joints were prepared as single side welded and double side welded by FSW. The welding was performed at various linear welding speeds (224, 560, and 900 mm/min) and one rotational speed of 710 rev./min using three different tool shapes. The results of macro examination and tensile test led to the selection of a single set of tool movement parameters at which the test joints for fatigue test were made. Samples were tested in two states of surface condition, i.e., "as-welded" and with mechanically removed marks left by rotating and moving tool during FSW process. Studies have shown that fatigue behavior of FSW joints depends on the tool shape used in the welding process as well as the surface condition of welded joints and manner of joint production.

  10. Effect of shot peening and grain refinement on the fatigue life and strength of commercially pure Al and two of its alloys: Al-2024-T3 and Al-7075-T6

    NASA Astrophysics Data System (ADS)

    Qandil, A.; Zaid, Adnan I. O.

    2016-08-01

    Aluminum and its alloys are widely used materials in automobile, aircraft and space craft industries due to their high strength- to- weight ratio and corrosion resistance beside their other useful properties. They are the second materials in use after steel alloys. Most of the failures in parts of aircrafts and space vehicles are mainly caused by fatigue and stress corrosion cracking. In this paper, the effect of shot peening on the fatigue life of commercially pure aluminumand two of its alloys namely:Al-2024 and Al-7075-T6 is presented and discussed. Furthermore, the effect of addition of vanadium to Al and Al grain refined by Ti and Ti+Bon Its fatigue life and strengthis also presented and discussed using scanning electron microscope, SEM. It was that shot peening and the addition of V toAl and Al onAl grain refined by Ti and Ti+B have resulted in enhancement of the fatigue life and strength. Ffinally, the effect of shot peening on the surface quality of the peened parts is also presented and discussed.

  11. Genomic and metabolic comparison with Dickeya dadantii 3937 reveals the emerging Dickeya solani potato pathogen to display distinctive metabolic activities and T5SS/T6SS-related toxin repertoire

    PubMed Central

    2014-01-01

    Background The pectinolytic enterobacteria of the Pectobacterium and Dickeya genera are causative agents of maceration-associated diseases affecting a wide variety of crops and ornamentals. For the past decade, the emergence of a novel species D. solani was observed in potato fields in Europe and the Mediterranean basin. The purpose of this study is to search by comparative genomics the genetic traits that could be distinctive to other Dickeya species and be involved in D. solani adaptation to the potato plant host. Results D. solani 3337 exhibits a 4.9 Mb circular genome that is characterized by a low content in mobile elements with the identification of only two full length insertion sequences. A genomic comparison with the deeply-annotated model D. dadantii 3937 strain was performed. While a large majority of Dickeya virulence genes are shared by both strains, a few hundreds genes of D. solani 3337, mostly regrouped in 25 genomic regions, are distinctive to D. dadantii 3937. These genomic regions are present in the other available draft genomes of D. solani strains and interestingly some of them were not found in the sequenced genomes of the other Dickeya species. These genomic regions regroup metabolic genes and are often accompanied by genes involved in transport systems. A metabolic analysis correlated some metabolic genes with distinctive functional traits of both D. solani 3337 and D. dadantii 3937. Three identified D. solani genomic regions also regroup NRPS/PKS encoding genes. In addition, D. solani encodes a distinctive arsenal of T5SS and T6SS-related toxin-antitoxin systems. These genes may contribute to bacteria-bacteria interactions and to the fitness of D. solani to the plant environment. Conclusions This study highlights the genomic specific traits of the emerging pathogen D. solani and will provide the basis for studying those that are involved in the successful adaptation of this emerging pathogen to the potato plant host. PMID:24735398

  12. Unfractionated spleen cells but not natural killer (NK) cells from RFM donors prevent the progression of host-versus-graft disease in murine RFM/(T6 x RFM)F1 chimeras.

    PubMed Central

    Hard, R C; Patel, M R; Keller, L M; Linna, T J

    1988-01-01

    Host-versus-graft (HVG) syndrome is the fatal allogenic disease which develops in susceptible strains of inbred mice following their perinatal inoculation with related F1 hybrid spleen cells. Deaths are caused by pathogenic immune complexes. It is thought that the antibody components of these complexes are produced by F1 donor B cells stimulated by the allogenic HVG reaction. To complement previous work that showed that lethal disease could be prevented if the HVG response was suppressed, the present studies tested whether or not it could also be prevented by augmenting HVG reactivity with the adoptive transfer of spleen cells syngenic with the host. The data show that unfractionated RFM spleen cells given on Days 13-14 prevented lethal disease in 86% of RFM/(T6 x RFM)F1 chimeras. Successful therapy was associated with the suppression of formation of nephropathic-immune complexes, and with the rejection of F1 donor cells or their gradual replacement by host cells. RFM spleen cells enriched for NK activity by a new improved method not only failed to prevent HVG disease but appeared to exacerbate it. This was also true of spleen cells that had been activated in vitro for 3 days with IL-2, a procedure that greatly enhanced their cytolytic activity against YAC-1 targets. It is suggested that therapy with NK cells failed, even after IL-2 activation, because they had no effect on proliferating and antibody-forming F1 donor cells that had engrafted in large numbers in the lymph nodes of the RFM hosts. PMID:3258280

  13. Mapping of the Pim-1 oncogene in mouse t-haplotypes and its use to define the relative map positions of the tcl loci t0(t6) and tw12 and the marker tf (tufted).

    PubMed

    Ark, B; Gummere, G; Bennett, D; Artzt, K

    1991-06-01

    Pim-1 is an oncogene activated in mouse T-cell lymphomas induced by Moloney and AKR mink cell focus (MCF) viruses. Pim-1 was previously mapped to chromosome 17 by somatic cell hybrids, and subsequently to the region between the hemoglobin alpha-chain pseudogene 4 (Hba-4ps) and the alpha-crystalline gene (Crya-1) by Southern blot analysis of DNA obtained from panels of recombinant inbred strains. We have now mapped Pim-1 more accurately in t-haplotypes by analysis of recombinant t-chromosomes. The recombinants were derived from Tts6tf/t12 parents backcrossed to + tf/ + tf, and scored for recombination between the loci of T and tf. For simplicity all t-complex lethal genes properly named tcl-tx are shortened to tx. The Pim-1 gene was localized 0.6 cM proximal to the tw12 lethal gene, thus placing the Pim-1 gene 5.2 cM distal to the H-2 region in t-haplotypes. Once mapped, the Pim-1 gene was used as a marker for further genetic analysis of t-haplotypes. tw12 is so close to tf that even with a large number of recombinants it was not possible to determine whether it is proximal or distal to tf. Southern blot analysis of DNA from T-tf recombinants with a separation of tw12 and tf indicated that tw12 is proximal to tf. The mapping of two allelic t-lethals, t0 and t6 with respect to tw12 and tf has also been a problem.(ABSTRACT TRUNCATED AT 250 WORDS)

  14. Bramson ORT Technical Institute. Report 92-T-6.

    ERIC Educational Resources Information Center

    Malan, Roland M.

    The New York State Comptroller's Office conducted an audit of Tuition Assistance Program (TAP) awards certified by Bramson ORT Technical Institute (BOTI) for the 1990-91 academic year, and performed a limited review of the Higher Education Services Corporation's (HESC's) records for the 1987-88 and 1989-90 academic years to detect errors or…

  15. Fatigue Strengths of Aircraft Materials: Axial-Load Fatigue Tests on Edge-Notched Sheet Specimens of 2024-T3 and 7075-T6 Aluminum Alloys and of SAE 4130 Steel with Notch Radii of 0.004 and 0.070 inch

    NASA Technical Reports Server (NTRS)

    Grover, H. J.; Hyler, W. S.; Jackson, L. R.

    1959-01-01

    The present report gives results of axial-load fatigue tests on notched specimens of three sheet materials: 2024-T3 and 7075-T6 aluminum alloys and normalized SAE 4130 steel. Two edge-notched specimens were designed and tested, each having a theoretical stress-concentration factor K(sub t) = 4.0. The radii of the notches were 0.004 and 0.070 inch. Tests of these specimens were run at two levels of nominal mean stress: 0 and 20,000 psi. Results of these studies extended information previously reported on tests of specimens with varying notch severity. They afford data on the variation of fatigue-strength reduction with notch radius and on the potential usefulness of Neuber's technical stress-concentration factor K(sub n).

  16. 7075-T6 and 2024-T351 Aluminum Alloy Fatigue Crack Growth Rate Data

    NASA Technical Reports Server (NTRS)

    Forth, Scott C.; Wright, Christopher W.; Johnston, William M., Jr.

    2005-01-01

    Experimental test procedures for the development of fatigue crack growth rate data has been standardized by the American Society for Testing and Materials. Over the past 30 years several gradual changes have been made to the standard without rigorous assessment of the affect these changes have on the precision or variability of the data generated. Therefore, the ASTM committee on fatigue crack growth has initiated an international round robin test program to assess the precision and variability of test results generated using the standard E647-00. Crack growth rate data presented in this report, in support of the ASTM roundrobin, shows excellent precision and repeatability.

  17. N = 4 supergravity for type IIB on T6/Bbb Z2 in presence of fluxes

    NASA Astrophysics Data System (ADS)

    Vaulà, Silvia

    2004-05-01

    We report on the construction of four-dimensional gauged supergravity models that can be interpreted as type IIB orientifold compactification in the presence of 3-form fluxes and D3-branes. We mainly address our attention to the symplectic embedding of the U-duality group of the theory and the consequent choice of the gauge group, whose four-dimensional Killing vectors are the remnants of the ten-dimensional fluxes. We briefly discuss the structure of the scalar potential arising from the gauging and the properties of the Killing vectors in order to preserve some amount of supersymmetry.

  18. Effects of Applied Load on 6061-T6 Aluminum Joined Employing a Novel Friction Bonding Process

    NASA Astrophysics Data System (ADS)

    Burkes, Douglas E.; Hallinan, Neil P.; Shropshire, Karen L.; Wells, Peter B.

    2008-12-01

    Friction bonding is under consideration for use in mass production of plate-type nuclear fuels for research reactors. This article discusses the effects of applied load (the most important process parameter for fabrication of these fuels) on temperature distribution, microstructure, and mechanical properties. Friction bonding experiments showed that tool geometry caused temperature gradients across the tool surface. Temperatures at the joint interface suggested the advancing side of the tool produced a majority of the frictional heat, while the retreating side of the tool mainly forged the plasticized material while bonding increased with applied load. The microstructure across the tool surface was also altered and, as a function of applied load, affected the mechanical properties. The 6061 aluminum alloy had mechanical properties close to a T4 temper after processing. Results documented in this article will aid in continual enhancement of friction bonding for nuclear fuel plate fabrication, and will hopefully contribute to continued advancement of friction stir welding (FSW) state of the art.

  19. Recreation Expenditures by U. S. Consumers, 1929-l979. Publication T-6-135.

    ERIC Educational Resources Information Center

    Kitchen, James W.; And Others

    This study explored trends in how Americans spent disposable personal income (DPI) on recreational activities from 1929 to 1979. Statistics were compiled from the Economic Report of the President and the United States Department of Commerce. Twelve classifications of recreation expenditures are presented: (1) books and maps; (2) magazines,…

  20. The effects of maintaining temperature in annealing heat treatment for an FSWed 6061-T6 Al alloy.

    PubMed

    Lee, Seung-Jun; Han, Min-Su; Kim, Seong-Jong

    2013-08-01

    The technological development of all kinds of lightweight transportation devices including vehicles, aircraft, ships, etc. has progressed markedly with the demand for energy saving and environmental protection. Aluminum alloy is in the spotlight as it is a suitable environmentally friendly material. However, deformation is a major problem during the welding process because aluminum alloy has a large thermal expansion coefficient. In addition, it is known that its corrosion resistance is excellent; nevertheless, in practice, considerable corrosion is generated and this is a major problem. To solve this problem, the friction stir welding (FSW) technology is applied extensively at various industrial fields as a new welding technique. This method involves a process in which materials are joined by frictional heat and physical force. Therefore, we evaluated improvements in mechanical properties and corrosion resistance through annealing heat treatment after FSW. The electrochemical experiment did not show a significant difference. However, the microstructure observation showed defectless, fine crystal particles, indicating excellent properties at 200-225°C.

  1. Prediction of low-cycle fatigue-life by acoustic emission—2: 7075-T6 aluminum alloy

    SciTech Connect

    Baram, J.; Rosen, M.

    1981-01-01

    Low-cycle fatigue tests were conducted by tension-compression until rupture, on a 2024-T3 aluminum alloy sheet. Initial crack sizes and orientations in the fatigue specimens were found to be randomly distributed. Acoustic emission was continuously monitored during the tests. Every few hundred cycles, the acoustic signal having the highest peak-amplitude, was recorded as an extremal event for the elapsed period. This high peak-amplitude is related to a fast crack propagation rate through a phenomenological relationship. The extremal peakamplitudes are shown by an ordered statistics treatment, to be extremally distributed. The statistical treatment enables the prediction of the number of cycles left until failure. Predictions performed a-posteriori based on results gained early in each fatigue test are in good agreement with actual fatigue lives. The amplitude distribution analysis of the acoustic signals emitted during cyclic stress appears to be a promising nondestructive method of predicting fatigue life.

  2. External And Internal Work Of A T-6 Paraplegic Propelling A Wheelchair And Arm Cranking A Cycle Ergometer: Case Study

    NASA Astrophysics Data System (ADS)

    Novak, Charles W.

    1982-02-01

    In this, the International Year of the Disabled, attention is directed among other areas toward rehabilitation and sports participation of wheelchair users. As an application of movement analysis in medicine and rehabilitation and as an application of sports research using biomechanics, this investigation was performed to compare the results of two methods of gathering data on the stress of wheelchair propelling at equivalent work loads and to account for differences in physiological responses with a mechanical analysis of wheelchair propelling. Physiological data collected were heart rate, systolic blood pressure, and rate-pressure product. A biomechanical cinematography analysis was used to determine external work in wheelchair propelling and to determine the extent to which modifications in segment actionsoccurred during increasing magnitude of work. A cycle ergometer was adjusted to replicate external work loads performed during wheelchair propelling. A t-test of equivalent external work loads indicated that heart rate was not different between the two exercise modes at the .05 level of significance. The t-test did indicate a significant difference in systolic blood pressure and rate-pressure product at the .05 level of significance. The biomechanical analysis of wheelchair propelling established that an increase in external work was accomplished by a decrease in the range of motion and an increase in the speed of movement. During cycle ergometry the range and speed of movement remained the same while resistance was increased. Results of the study established that while heart rate for equivalent external work loads was the same for wheelchair propelling and arm cranking cycle ergometry, systolic blood pressure and rate-pressure product were not the same. The suggestion was that some means of propelling a wheelchair other than that which is con-sidered "standard" might be considered which produces less stressful responses in wheelchair users.

  3. The effects of maintaining temperature in annealing heat treatment for an FSWed 6061-T6 Al alloy.

    PubMed

    Lee, Seung-Jun; Han, Min-Su; Kim, Seong-Jong

    2013-08-01

    The technological development of all kinds of lightweight transportation devices including vehicles, aircraft, ships, etc. has progressed markedly with the demand for energy saving and environmental protection. Aluminum alloy is in the spotlight as it is a suitable environmentally friendly material. However, deformation is a major problem during the welding process because aluminum alloy has a large thermal expansion coefficient. In addition, it is known that its corrosion resistance is excellent; nevertheless, in practice, considerable corrosion is generated and this is a major problem. To solve this problem, the friction stir welding (FSW) technology is applied extensively at various industrial fields as a new welding technique. This method involves a process in which materials are joined by frictional heat and physical force. Therefore, we evaluated improvements in mechanical properties and corrosion resistance through annealing heat treatment after FSW. The electrochemical experiment did not show a significant difference. However, the microstructure observation showed defectless, fine crystal particles, indicating excellent properties at 200-225°C. PMID:23920178

  4. A damage tolerance comparison of 7075-T6 aluminum alloy and IM7/977-2 carbon/epoxy

    NASA Technical Reports Server (NTRS)

    Nettles, Alan T.; Lance, David G.; Hodge, Andrew J.

    1991-01-01

    A comparison of low velocity impact damage between one of the strongest aluminum alloys, to a new, damage tolerant resin system as a matrix for high strength carbon fibers was examined in this study. The aluminum and composite materials were used as face sheets on a 0.13 g/cu cm aluminum honeycomb. Four levels of impact energy were used; 2.6 J, 5.3 J, 7.8 J and 9.9 J. The beams were compared for static strength and fatique life by use of the four-point bend flexure test. It was found that in the undamaged state the specific strength of the composite face sheets was about twice that of the aluminum face sheets. A sharp drop in strength was observed for the composite specimens impacted at the lowest (2.6J) energy level, but the overall specific strength was still higher than for the aluminum specimens. At all impact energy levels tested, the static specific strength of the composite face sheets were significantly higher than the aluminum face sheets. The fatigue life of the most severely damaged composite specimen was about 17 times greater than the undamaged aluminum specimens when cycled at 1 Hz between 20 percent and 85 percent of ultimate breaking load.

  5. Draft Genome Sequence of Pseudomonas azotifigens Strain DSM 17556T (6H33bT), a Nitrogen Fixer Strain Isolated from a Compost Pile

    PubMed Central

    Busquets, Antonio; Peña, Arantxa; Gomila, Margarita; Mulet, Magdalena; Mayol, Joan; García-Valdés, Elena; Bennasar, Antonio; Huntemann, Marcel; Han, James; Chen, I-Min; Mavromatis, Konstantinos; Markowitz, Victor; Palaniappan, Krishnaveni; Ivanova, Natalia; Schaumberg, Andrew; Pati, Amrita; Reddy, T. B. K.; Nordberg, Henrik; Woyke, Tanja; Klenk, Hans-Peter; Kyrpides, Nikos

    2013-01-01

    Pseudomonas azotifigens strain 6H33bT is a nitrogen fixer isolated from a hyperthermal compost pile in 2005 by Hatayama and collaborators. Here we report the draft genome, which has an estimated size of 5.0 Mb, exhibits an average G+C content of 66.73%, and is predicted to encode 4,536 protein-coding genes and 100 RNA genes. PMID:24179119

  6. 22q11.2 Deletion Syndrome due to a Translocation t(6;22) in a Patient Conceived via in vitro Fertilization

    PubMed Central

    Gollo Dantas, Anelisa; Bortolai, Adriana; Moysés-Oliveira, Mariana; Takeno Herrero, Sylvia; Azoubel Antunes, Adriana; Tavares Costa-Carvalho, Beatriz; Ayres Meloni, Vera; Melaragno, Maria Isabel

    2016-01-01

    We report on a patient conceived via in vitro fertilization (IVF) with a 22q11.2 deletion due to an unusual unbalanced translocation involving chromosomes 6 and 22 in a karyotype with 45 chromosomes. Cytogenomic studies showed that the patient has a 3.3-Mb deletion of chromosome 22q and a 0.4-Mb deletion of chromosome 6p, which resulted in haploinsufficiency of the genes responsible for the 22q11.2 deletion syndrome and also of the IRF4 gene, a member of the interferon regulatory factor family of transcription factors, which is expressed in the immune system cells. The rearrangement could be due to the manipulation of the embryo or as a sporadic event unrelated to IVF. Translocation involving chromosome 22 in a karyotype with 45 chromosomes is a rare event, with no previous reports involving chromosomes 6p and 22q. PMID:26997945

  7. Spin Forming of an Aluminum 2219-T6 Aft Bulkhead for the Orion Multi-Purpose Crew Vehicle: Phase II Supplemental Report

    NASA Technical Reports Server (NTRS)

    Piascik, Robert S.; Squire, Michael D.; Domack, Marcia S.; Hoffman, Eric K.

    2015-01-01

    The principal focus of this project was to assist the Orion Multi-Purpose Crew Vehicle (MPCV) Program in developing a spin forming fabrication process for manufacture of the aft bulkhead of the pressure vessel. The spin forming process will enable a single piece aluminum (Al) 2219 aft bulkhead which will eliminate the current multiple piece welded construction, simplify fabrication, and lead to an enhanced design that will reduce vehicle weight by eliminating welds. Phase I of this assessment explored spin forming the single-piece forward pressure vessel bulkhead from aluminum-lithium 2195.

  8. Experimental Investigation of Effects of Random Loading on the Fatigue Life of Notched Cantilever-Beam Specimens of 7075-T6 Aluminum Alloy

    NASA Technical Reports Server (NTRS)

    Fralich, Robert W.

    1959-01-01

    Results of random-loading fatigue tests on 125 notched cantilever-beam specimens and constant-amplitude fatigue tests on 46 similar specimens are presented in terms of the root-mean-square value of peak stresses. The results from the two sets of tests are compared on the basis of time to failure, where the results from the constant-amplitude tests are expressed in terms of an equivalent time to failure based on the natural period of vibration. Compared on this basis, failure at the lower stress levels occurred in a shorter time for the random loading than for the constant-amplitude loading, whereas at the higher stress levels failure occurred in the reverse order. A theoretical result for random loading is also presented and compared with the experimental results. The theoretical result shows good agreement with experiment for low values of stress but underestimates the time to failure at the higher stresses.

  9. Practical aspects involved in the design and set up of a 0.15 T, 6-coil resistive magnet, whole body NMR imaging facility.

    PubMed

    Thomas, S R; Ackerman, J L; Kereiakes, J G

    1984-01-01

    Many technical and logistical questions must be addressed when planning the installation of an NMR imaging system. These considerations become particularly significant when the facility is being established within an existing medical center complex. This paper presents a report on the practical aspects and experience obtained in siting a 6-coil 0.15 T resistive magnet system. The topics discussed include: floor loading; ferromagnetic environment; the effect of iron on the magnet field strength and homogeneity characteristics; shimming procedures; temperature stability requirements; rf shielding; and effects of the magnetic field on common medical instrumentation and magnetic media. It was found that the field shift as a function of the distance of a steel mass from the center of the magnet exhibited an (1/r)5.2 +/- 0.5 to (1/r) 4.2 +/- 0.3 dependence for axial and radial positions respectively which, as expected, is somewhat weaker than the (1/r)6 dependence expected by point dipole approximations. Field distortions caused by the presence of ferromagnetic material in radial positions may be essentially fully compensated with first order transverse shim coils (most conveniently, the x and y imaging gradient coils could be used). Axially distributed material requires, in addition to first order z-gradient correction, higher order axial shim compensation. The temperature stability of the magnet system over the scan period must be better than 0.2 degrees C to insure that temperature-induced field fluctuations are less than the intrinsic static inhomogeneity: and, ideally, below 0.01 degrees C to reduce these fluctuations to less than those caused by power supply instability.

  10. Cytotoxic and antiproliferative effects induced by a non terpenoid polar extract of A. indica seeds on 3T6 murine fibroblasts in culture.

    PubMed

    Di Ilio, Vincenzo; Pasquariello, Nicoletta; van der Esch, Andrew S; Cristofaro, Massimo; Scarsella, Gianfranco; Risuleo, Gianfranco

    2006-07-01

    Neem oil is a natural product obtained from the seeds of the tree Azadirachta indica. Its composition is very complex and the oil exhibits a number of biological activities. The most studied component is the terpenoid azadirachtin which is used for its insecticidal and putative antimicrobial properties. In this report we investigate the biological activity of partially purified components of the oil obtained from A. indica. We show that the semi-purified fractions have moderate to strong cytotoxicity. However, this is not attributable to azadirachtin but to other active compounds present in the mixture. Each fraction was further purified by appropriate extraction procedures and we observed a differential cytotoxicity in the various sub-fractions. This led us to investigate the mode of cell death. After treatment with the oil fractions we observed positivity to TUNEL staining and extensive internucleosomal DNA degradation both indicating apoptotic death. The anti-proliferative properties of the neem oil-derived compounds were also assayed by evaluation of the nuclear PCNA levels (Proliferating Cell Nuclear Antigen). PCNA is significantly reduced in cells treated with a specific fraction of neem oil. Finally, our results strongly suggest a possible involvement of the mitochondrial pathway in the apoptotic death. PMID:16652209

  11. Established Microbial Colonies Can Survive Type VI Secretion Assault

    PubMed Central

    Borenstein, David Bruce; Ringel, Peter; Basler, Marek; Wingreen, Ned S.

    2015-01-01

    Type VI secretion (T6S) is a cell-to-cell injection system that can be used as a microbial weapon. T6S kills vulnerable cells, and is present in close to 25% of sequenced Gram-negative bacteria. To examine the ecological role of T6S among bacteria, we competed self-immune T6S+ cells and T6S-sensitive cells in simulated range expansions. As killing takes place only at the interface between sensitive and T6S+ strains, while growth takes place everywhere, sufficiently large domains of sensitive cells can achieve net growth in the face of attack. Indeed T6S-sensitive cells can often outgrow their T6S+ competitors. We validated these findings through in vivo competition experiments between T6S+ Vibrio cholerae and T6S-sensitive Escherichia coli. We found that E. coli can survive and even dominate so long as they have an adequate opportunity to form microcolonies at the outset of the competition. Finally, in simulated competitions between two equivalent and mutually sensitive T6S+ strains, the more numerous strain has an advantage that increases with the T6S attack rate. We conclude that sufficiently large domains of T6S-sensitive individuals can survive attack and potentially outcompete self-immune T6S+ bacteria. PMID:26485125

  12. Only one of the two type VI secretion systems encoded in the Salmonella enterica serotype Dublin genome is involved in colonization of the avian and murine hosts

    PubMed Central

    2014-01-01

    The type VI secretion system (T6SS) is a virulence factor for many Gram-negative bacteria. Salmonella genus harbors five phylogenetically distinct T6SS loci encoded in Salmonella Pathogenicity Islands (SPIs) SPI-6, SPI-19, SPI-20, SPI-21 and SPI-22, which are differentially distributed among serotypes. The T6SSs encoded in SPI-6 and SPI-19 contribute to pathogenesis of serotypes Typhimurium and Gallinarum in mice and chickens, respectively. Salmonella Dublin is a pathogen restricted to cattle where it causes a systemic disease. Also, it can colonize other hosts such as chickens and mice, which can act as reservoirs of this serotype. Salmonella Dublin harbors the genes for both T6SSSPI-6 and T6SSSPI-19. This study has determined the contribution of T6SSSPI-6 and T6SSSPI-19 to host-colonization by Salmonella Dublin using avian and murine models of infection. Competitive index experiments showed that, a mutant strain lacking both T6SSs (∆T6SSSPI-6/∆T6SSSPI-19) presents a strong colonization defect in cecum of chickens, similar to the defect observed for the ∆T6SSSPI-6 mutant, suggesting that this serotype requires a functional T6SSSPI-6 for efficient colonization of the avian gastrointestinal tract. Colonization of mice was also defective, although to a lesser extent than in chickens. In contrast, the T6SSSPI-19 was not necessary for colonization of either chickens or mice. Transfer of T6SSSPI-6, but not T6SSSPI-19, restored the ability of the double mutant to colonize both animal hosts. Our data indicate that Salmonella Dublin requires only the T6SSSPI-6 for efficient colonization of mice and chickens, and that the T6SSSPI-6 and T6SSSPI-19 are not functionally redundant. PMID:24405577

  13. 49 CFR Appendix C to Part 180 - Eddy Current Examination With Visual Inspection for DOT 3AL Cylinders Manufactured of Aluminum...

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... for DOT 3AL Cylinders Manufactured of Aluminum Alloy 6351-T6 C Appendix C to Part 180 Transportation... Cylinders Manufactured of Aluminum Alloy 6351-T6 1. Examination Procedure. Each facility performing eddy... ring and probe for each DOT-3AL cylinder manufactured of aluminum alloy 6351-T6 to be inspected must...

  14. 49 CFR Appendix C to Part 180 - Eddy Current Examination With Visual Inspection for DOT 3AL Cylinders Manufactured of Aluminum...

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... for DOT 3AL Cylinders Manufactured of Aluminum Alloy 6351-T6 C Appendix C to Part 180 Transportation... Cylinders Manufactured of Aluminum Alloy 6351-T6 1. Examination Procedure. Each facility performing eddy... ring and probe for each DOT-3AL cylinder manufactured of aluminum alloy 6351-T6 to be inspected must...

  15. 49 CFR Appendix C to Part 180 - Eddy Current Examination With Visual Inspection for DOT 3AL Cylinders Manufactured of Aluminum...

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... for DOT 3AL Cylinders Manufactured of Aluminum Alloy 6351-T6 C Appendix C to Part 180 Transportation... Cylinders Manufactured of Aluminum Alloy 6351-T6 1. Examination Procedure. Each facility performing eddy... ring and probe for each DOT-3AL cylinder manufactured of aluminum alloy 6351-T6 to be inspected must...

  16. 49 CFR Appendix C to Part 180 - Eddy Current Examination With Visual Inspection for DOT 3AL Cylinders Manufactured of Aluminum...

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... for DOT 3AL Cylinders Manufactured of Aluminum Alloy 6351-T6 C Appendix C to Part 180 Transportation... Cylinders Manufactured of Aluminum Alloy 6351-T6 1. Examination Procedure. Each facility performing eddy... ring and probe for each DOT-3AL cylinder manufactured of aluminum alloy 6351-T6 to be inspected must...

  17. 49 CFR Appendix C to Part 180 - Eddy Current Examination With Visual Inspection for DOT 3AL Cylinders Manufactured of Aluminum...

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... for DOT 3AL Cylinders Manufactured of Aluminum Alloy 6351-T6 C Appendix C to Part 180 Transportation... Cylinders Manufactured of Aluminum Alloy 6351-T6 1. Examination Procedure. Each facility performing eddy... ring and probe for each DOT-3AL cylinder manufactured of aluminum alloy 6351-T6 to be inspected must...

  18. The Type VI Secretion System Encoded in SPI-6 Plays a Role in Gastrointestinal Colonization and Systemic Spread of Salmonella enterica serovar Typhimurium in the Chicken

    PubMed Central

    Pezoa, David; Yang, Hee-Jeong; Blondel, Carlos J.; Santiviago, Carlos A.; Andrews-Polymenis, Helene L.; Contreras, Inés

    2013-01-01

    The role of the Salmonella Pathogenicity Islands (SPIs) in pathogenesis of Salmonella enterica Typhimurium infection in the chicken is poorly studied, while many studies have been completed in murine models. The Type VI Secretion System (T6SS) is a recently described protein secretion system in Gram-negative bacteria. The genus Salmonella contains five phylogenetically distinct T6SS encoded in differentially distributed genomic islands. S. Typhimurium harbors a T6SS encoded in SPI-6 (T6SSSPI-6), which contributes to the ability of Salmonella to colonize mice. On the other hand, serotype Gallinarum harbors a T6SS encoded in SPI-19 (T6SSSPI-19) that is required for colonization of chicks. In this work, we investigated the role of T6SSSPI-6 in infection of chicks by S. Typhimurium. Oral infection of White Leghorn chicks showed that a ΔT6SSSPI-6 mutant had reduced colonization of the gut and internal organs, compared with the wild-type strain. Transfer of the intact T6SSSPI-6 gene cluster into the T6SS mutant restored bacterial colonization. In addition, our results showed that transfer of T6SSSPI-19 from S. Gallinarum to the ΔT6SSSPI-6 mutant of S. Typhimurium not only complemented the colonization defect but also resulted in a transient increase in the colonization of the cecum and ileum of chicks at days 1 and 3 post-infection. Our data indicates that T6SSSPI-6 contributes to chicken colonization and suggests that both T6SSSPI-6 and T6SSSPI-19 perform similar functions in vivo despite belonging to different phylogenetic families. PMID:23691117

  19. Marker for type VI secretion system effectors

    PubMed Central

    Salomon, Dor; Kinch, Lisa N.; Trudgian, David C.; Guo, Xiaofeng; Klimko, John A.; Grishin, Nick V.; Mirzaei, Hamid; Orth, Kim

    2014-01-01

    Bacteria use diverse mechanisms to kill, manipulate, and compete with other cells. The recently discovered type VI secretion system (T6SS) is widespread in bacterial pathogens and used to deliver virulence effector proteins into target cells. Using comparative proteomics, we identified two previously unidentified T6SS effectors that contained a conserved motif. Bioinformatic analyses revealed that this N-terminal motif, named MIX (marker for type six effectors), is found in numerous polymorphic bacterial proteins that are primarily located in the T6SS genome neighborhood. We demonstrate that several MIX-containing proteins are T6SS effectors and that they are not required for T6SS activity. Thus, we propose that MIX-containing proteins are T6SS effectors. Our findings allow for the identification of numerous uncharacterized T6SS effectors that will undoubtedly lead to the discovery of new biological mechanisms. PMID:24927539

  20. Only one of the two type VI secretion systems encoded in the Salmonella enterica serotype Dublin genome is involved in colonization of the avian and murine hosts.

    PubMed

    Pezoa, David; Blondel, Carlos J; Silva, Cecilia A; Yang, Hee-Jeong; Andrews-Polymenis, Helene; Santiviago, Carlos A; Contreras, Inés

    2014-01-09

    The type VI secretion system (T6SS) is a virulence factor for many Gram-negative bacteria. Salmonella genus harbors five phylogenetically distinct T6SS loci encoded in Salmonella Pathogenicity Islands (SPIs) SPI-6, SPI-19, SPI-20, SPI-21 and SPI-22, which are differentially distributed among serotypes. The T6SSs encoded in SPI-6 and SPI-19 contribute to pathogenesis of serotypes Typhimurium and Gallinarum in mice and chickens, respectively. Salmonella Dublin is a pathogen restricted to cattle where it causes a systemic disease. Also, it can colonize other hosts such as chickens and mice, which can act as reservoirs of this serotype. Salmonella Dublin harbors the genes for both T6SS(SPI-6) and T6SS(SPI-19). This study has determined the contribution of T6SS(SPI-6) and T6SS(SPI-19) to host-colonization by Salmonella Dublin using avian and murine models of infection. Competitive index experiments showed that, a mutant strain lacking both T6SSs (∆T6SS(SPI-6)/∆T6SS(SPI-19)) presents a strong colonization defect in cecum of chickens, similar to the defect observed for the ∆T6SS(SPI-6) mutant, suggesting that this serotype requires a functional T6SS(SPI-6) for efficient colonization of the avian gastrointestinal tract. Colonization of mice was also defective, although to a lesser extent than in chickens. In contrast, the T6SS(SPI-19) was not necessary for colonization of either chickens or mice. Transfer of T6SS(SPI-6), but not T6SS(SPI-19), restored the ability of the double mutant to colonize both animal hosts. Our data indicate that Salmonella Dublin requires only the T6SS(SPI-6) for efficient colonization of mice and chickens, and that the T6SS(SPI-6) and T6SS(SPI-19) are not functionally redundant.

  1. Altering trehalose-6-phosphate content in transgenic potato tubers affects tuber growth and alters responsiveness to hormones during sprouting.

    PubMed

    Debast, Stefan; Nunes-Nesi, Adriano; Hajirezaei, Mohammad R; Hofmann, Jörg; Sonnewald, Uwe; Fernie, Alisdair R; Börnke, Frederik

    2011-08-01

    Trehalose-6-phosphate (T6P) is a signaling metabolite that regulates carbon metabolism, developmental processes, and growth in plants. In Arabidopsis (Arabidopsis thaliana), T6P signaling is, at least in part, mediated through inhibition of the SNF1-related protein kinase SnRK1. To investigate the role of T6P signaling in a heterotrophic, starch-accumulating storage organ, transgenic potato (Solanum tuberosum) plants with altered T6P levels specifically in their tubers were generated. Transgenic lines with elevated T6P levels (B33-TPS, expressing Escherichia coli osmoregulatory trehalose synthesis A [OtsA], which encodes a T6P synthase) displayed reduced starch content, decreased ATP contents, and increased respiration rate diagnostic for high metabolic activity. On the other hand, lines with significantly reduced T6P (B33-TPP, expressing E. coli OtsB, which encodes a T6P phosphatase) showed accumulation of soluble carbohydrates, hexose phosphates, and ATP, no change in starch when calculated on a fresh weight basis, and a strongly reduced tuber yield. [¹⁴C]glucose feeding to transgenic tubers indicated that carbon partitioning between starch and soluble carbohydrates was not altered. Transcriptional profiling of B33-TPP tubers revealed that target genes of SnRK1 were strongly up-regulated and that T6P inhibited potato tuber SnRK1 activity in vitro. Among the SnRK1 target genes in B33-TPP tubers, those involved in the promotion of cell proliferation and growth were down-regulated, while an inhibitor of cell cycle progression was up-regulated. T6P-accumulating tubers were strongly delayed in sprouting, while those with reduced T6P sprouted earlier than the wild type. Early sprouting of B33-TPP tubers correlated with a reduced abscisic acid content. Collectively, our data indicate that T6P plays an important role for potato tuber growth.

  2. Deformation and Damage of Two Aluminum Alloys from Ballistic Impact

    NASA Astrophysics Data System (ADS)

    Anderson, Charles; Dannemann, Kathryn

    2001-06-01

    A series of impact experiments was conducted on 4.75-mm-thick aluminum plates to investigate the deformation and damage behavior of two aluminum alloys, 6061-T6 and 7075-T6. The projectile used was the Sierra 165 lead-filled bullet. Impact velocities were varied from approximately 260 m/s to 370 m/s, with the higher velocities just slightly below the ballistic limit velocities for the plates. The flow stress for 7075-T6 aluminum is approximately 50than it is for 6061-T6 aluminum; however, the fracture toughness of the 7075-T6 alloy is considerably lower than that for 6061-T6. A grid pattern was placed on the rear surface of the plates, and post-test analysis determined the in-plane back-surface strains. The 7075-T6 aluminum plates have less deformation than the 6061-T6 plates at the same impact velocity, but at some critical velocity, a through-thickness crack appears in the plate, ultimately leading to bullet perforation. In contrast, the 6061-T6 alloy continues to deform and fails by ductile tearing. These differences in damage/failure result in the two alloys having much closer ballistic limit velocities than expected based on the differences in strength.

  3. Bile Salts Modulate the Mucin-Activated Type VI Secretion System of Pandemic Vibrio cholerae

    PubMed Central

    Unterweger, Daniel; Diaz-Satizabal, Laura; Ogg, Stephen; Pukatzki, Stefan

    2015-01-01

    The causative agent of cholera, Vibrio cholerae, regulates its diverse virulence factors to thrive in the human small intestine and environmental reservoirs. Among this pathogen’s arsenal of virulence factors is the tightly regulated type VI secretion system (T6SS). This system acts as an inverted bacteriophage to inject toxins into competing bacteria and eukaryotic phagocytes. V. cholerae strains responsible for the current 7th pandemic activate their T6SS within the host. We established that T6SS-mediated competition occurs upon T6SS activation in the infant mouse, and that this system is functional under anaerobic conditions. When investigating the intestinal host factors mucins (a glycoprotein component of mucus) and bile for potential regulatory roles in controlling the T6SS, we discovered that once mucins activate the T6SS, bile acids can further modulate T6SS activity. Microbiota modify bile acids to inhibit T6SS-mediated killing of commensal bacteria. This interplay is a novel interaction between commensal bacteria, host factors, and the V. cholerae T6SS, showing an active host role in infection. PMID:26317760

  4. Bile Salts Modulate the Mucin-Activated Type VI Secretion System of Pandemic Vibrio cholerae.

    PubMed

    Bachmann, Verena; Kostiuk, Benjamin; Unterweger, Daniel; Diaz-Satizabal, Laura; Ogg, Stephen; Pukatzki, Stefan

    2015-01-01

    The causative agent of cholera, Vibrio cholerae, regulates its diverse virulence factors to thrive in the human small intestine and environmental reservoirs. Among this pathogen's arsenal of virulence factors is the tightly regulated type VI secretion system (T6SS). This system acts as an inverted bacteriophage to inject toxins into competing bacteria and eukaryotic phagocytes. V. cholerae strains responsible for the current 7th pandemic activate their T6SS within the host. We established that T6SS-mediated competition occurs upon T6SS activation in the infant mouse, and that this system is functional under anaerobic conditions. When investigating the intestinal host factors mucins (a glycoprotein component of mucus) and bile for potential regulatory roles in controlling the T6SS, we discovered that once mucins activate the T6SS, bile acids can further modulate T6SS activity. Microbiota modify bile acids to inhibit T6SS-mediated killing of commensal bacteria. This interplay is a novel interaction between commensal bacteria, host factors, and the V. cholerae T6SS, showing an active host role in infection.

  5. The type VI secretion system gene cluster of Salmonella typhimurium: required for full virulence in mice.

    PubMed

    Liu, Ji; Guo, Ji-Tao; Li, Yong-Guo; Johnston, Randal N; Liu, Gui-Rong; Liu, Shu-Lin

    2013-07-01

    Type VI secretion system (T6SS) has increasingly been believed to participate in the infection process for many bacterial pathogens, but its role in the virulence of Salmonella typhimurium remains unclear. To look into this, we deleted the T6SS cluster from the genome of S. typhimurium 14028s and analyzed the phenotype of the resulting T6SS knockout mutant (T6SSKO mutant) in vitro and in vivo. We found that the T6SSKO mutant exhibited reduced capability in colonizing the spleen and liver in an in vivo colonization competition model in BALB/c mice infected by the oral route. Additionally, infection via intraperitoneal administration also showed that the T6SSKO mutant was less capable of colonizing the mouse spleen and liver than the wild-type strain. We did not detect significant differences between the T6SSKO and wild-type strains in epithelial cell invasion tests. However, in the macrophage RAW264.7 cell line, the T6SSKO mutant survived and proliferated significantly more poorly than the wild-type strain. These findings indicate that T6SS gene cluster is required for full virulence of S. typhimurium 14028s in BALB/c mice, possibly due to its roles in bacterial survival and proliferation in macrophages.

  6. Genetic Dissection of the Type VI Secretion System in Acinetobacter and Identification of a Novel Peptidoglycan Hydrolase, TagX, Required for Its Biogenesis

    PubMed Central

    Weber, Brent S.; Hennon, Seth W.; Wright, Meredith S.; Scott, Nichollas E.; de Berardinis, Véronique; Foster, Leonard J.; Ayala, Juan A.; Adams, Mark D.

    2016-01-01

    ABSTRACT The type VI secretion system (T6SS) is a widespread secretory apparatus produced by Gram-negative bacteria that has emerged as a potent mediator of antibacterial activity during interbacterial interactions. Most Acinetobacter species produce a genetically conserved T6SS, although the expression and functionality of this system vary among different strains. Some pathogenic Acinetobacter baumannii strains activate this secretion system via the spontaneous loss of a plasmid carrying T6SS repressors. In this work, we compared the expression of T6SS-related genes via transcriptome sequencing and differential proteomics in cells with and without the plasmid. This approach, together with the mutational analysis of the T6SS clusters, led to the determination of the genetic components required to elaborate a functional T6SS in the nosocomial pathogen A. baumannii and the nonpathogen A. baylyi. By constructing a comprehensive combination of mutants with changes in the T6SS-associated vgrG genes, we delineated their relative contributions to T6SS function. We further determined the importance of two effectors, including an effector-immunity pair, for antibacterial activity. Our genetic analysis led to the identification of an essential membrane-associated structural component named TagX, which we have characterized as a peptidoglycan hydrolase possessing l,d-endopeptidase activity. TagX shows homology to known bacteriophage l,d-endopeptidases and is conserved in the T6SS clusters of several bacterial species. We propose that TagX is the first identified enzyme that fulfills the important role of enabling the transit of T6SS machinery across the peptidoglycan layer of the T6SS-producing bacterium. PMID:27729508

  7. The archetype Pseudomonas aeruginosa proteins TssB and TagJ form a novel subcomplex in the bacterial type VI secretion system.

    PubMed

    Lossi, Nadine S; Manoli, Eleni; Simpson, Pete; Jones, Cerith; Hui, Kailyn; Dajani, Rana; Coulthurst, Sarah J; Freemont, Paul; Filloux, Alain

    2012-10-01

    In Pseudomonas aeruginosa three type VI secretion systems (T6SSs) coexist, called H1- to H3-T6SSs. Several T6SS components are proposed to be part of a macromolecular complex resembling the bacteriophage tail. The T6SS protein HsiE1 (TagJ) is unique to the H1-T6SS and absent from the H2- and H3-T6SSs. We demonstrate that HsiE1 interacts with a predicted N-terminal α-helix in HsiB1 (TssB) thus forming a novel subcomplex of the T6SS. HsiB1 is homologous to the Vibrio cholerae VipA component, which contributes to the formation of a bacteriophage tail sheath-like structure. We show that the interaction between HsiE1 and HsiB1 is specific and does not occur between HsiE1 and HsiB2. Proteins of the TssB family encoded in T6SS clusters lacking a gene encoding a TagJ-like component are often devoid of the predicted N-terminal helical region, which suggests co-evolution. We observe that a synthetic peptide corresponding to the N-terminal 20 amino acids of HsiB1 interacts with purified HsiE1 protein. This interaction is a common feature to other bacterial T6SSs that display a TagJ homologue as shown here with Serratia marcescens. We further show that hsiE1 is a non-essential gene for the T6SS and suggest that HsiE1 may modulate incorporation of HsiB1 into the T6SS.

  8. Constitutive type VI secretion system expression gives Vibrio cholerae intra- and interspecific competitive advantages.

    PubMed

    Unterweger, Daniel; Kitaoka, Maya; Miyata, Sarah T; Bachmann, Verena; Brooks, Teresa M; Moloney, Jessica; Sosa, Oscar; Silva, David; Duran-Gonzalez, Jorge; Provenzano, Daniele; Pukatzki, Stefan

    2012-01-01

    The type VI secretion system (T6SS) mediates protein translocation across the cell membrane of Gram-negative bacteria, including Vibrio cholerae - the causative agent of cholera. All V. cholerae strains examined to date harbor gene clusters encoding a T6SS. Structural similarity and sequence homology between components of the T6SS and the T4 bacteriophage cell-puncturing device suggest that the T6SS functions as a contractile molecular syringe to inject effector molecules into prokaryotic and eukaryotic target cells. Regulation of the T6SS is critical. A subset of V. cholerae strains, including the clinical O37 serogroup strain V52, express T6SS constitutively. In contrast, pandemic strains impose tight control that can be genetically disrupted: mutations in the quorum sensing gene luxO and the newly described regulator gene tsrA lead to constitutive T6SS expression in the El Tor strain C6706. In this report, we examined environmental V. cholerae isolates from the Rio Grande with regard to T6SS regulation. Rough V. cholerae lacking O-antigen carried a nonsense mutation in the gene encoding the global T6SS regulator VasH and did not display virulent behavior towards Escherichia coli and other environmental bacteria. In contrast, smooth V. cholerae strains engaged constitutively in type VI-mediated secretion and displayed virulence towards prokaryotes (E. coli and other environmental bacteria) and a eukaryote (the social amoeba Dictyostelium discoideum). Furthermore, smooth V. cholerae strains were able to outcompete each other in a T6SS-dependent manner. The work presented here suggests that constitutive T6SS expression provides V. cholerae with an advantage in intraspecific and interspecific competition.

  9. The trehalose 6-phosphate/SnRK1 signaling pathway primes growth recovery following relief of sink limitation.

    PubMed

    Nunes, Cátia; O'Hara, Liam E; Primavesi, Lucia F; Delatte, Thierry L; Schluepmann, Henriette; Somsen, Govert W; Silva, Anabela B; Fevereiro, Pedro S; Wingler, Astrid; Paul, Matthew J

    2013-07-01

    Trehalose 6-P (T6P) is a sugar signal in plants that inhibits SNF1-related protein kinase, SnRK1, thereby altering gene expression and promoting growth processes. This provides a model for the regulation of growth by sugar. However, it is not known how this model operates under sink-limited conditions when tissue sugar content is uncoupled from growth. To test the physiological importance of this model, T6P, SnRK1 activities, sugars, gene expression, and growth were measured in Arabidopsis (Arabidopsis thaliana) seedlings after transfer to cold or zero nitrogen compared with sugar feeding under optimal conditions. Maximum in vitro activities of SnRK1 changed little, but T6P accumulated up to 55-fold, correlating with tissue Suc content in all treatments. SnRK1-induced and -repressed marker gene expression strongly related to T6P above and below a threshold of 0.3 to 0.5 nmol T6P g(-1) fresh weight close to the dissociation constant (4 µm) of the T6P/ SnRK1 complex. This occurred irrespective of the growth response to Suc. This implies that T6P is not a growth signal per se, but through SnRK1, T6P primes gene expression for growth in response to Suc accumulation under sink-limited conditions. To test this hypothesis, plants with genetically decreased T6P content and SnRK1 overexpression were transferred from cold to warm to analyze the role of T6P/SnRK1 in relief of growth restriction. Compared with the wild type, these plants were impaired in immediate growth recovery. It is concluded that the T6P/SnRK1 signaling pathway responds to Suc induced by sink restriction that enables growth recovery following relief of limitations such as low temperature.

  10. The Versatile Type VI Secretion System

    PubMed Central

    Alteri, Christopher J.; Mobley, Harry L.T.

    2016-01-01

    Summary Bacterial Type VI Secretion Systems (T6SS) function as contractile nanomachines to puncture target cells and deliver lethal effectors. In the ten years since the discovery of the T6SS, much has been learned about the structure and function of this versatile protein secretion apparatus. Most of the conserved protein components that comprise the T6SS apparatus itself have been identified and ascribed specific functions. In addition, numerous effector proteins that are translocated by the T6SS have been identified and characterized. These protein effectors usually represent toxic cargoes that are delivered by the attacker cell to a target cell. The field is beginning to better understand the lifestyle or physiology that dictates when bacteria normally express their T6SS. In this Chapter, we consider what is known about the structure and regulation of the T6SS, the numerous classes of antibacterial effector T6SS substrates, and how the action of the T6SS relates to a given lifestyle or behavior in certain bacteria. PMID:27227310

  11. Deformation and Damage of Two Aluminum Alloys from Ballistic Impact

    NASA Astrophysics Data System (ADS)

    Anderson, Charles E., Jr.; Dannemann, Kathryn A.

    2002-07-01

    A series of impact experiments were conducted on 4.76-mm-thick aluminum plates to investigate the deformation and damage behavior of two aluminum alloys, 6061-T6 and 7075-T6. The Sierra 165 lead-filled bullet was used to load the plates. Impact velocities were varied from approximately 260 m/s to 370 m/s. The flow stress for 7075-T6 aluminum is approximately twice that for 6061-T6 aluminum; however, the ballistic limit velocities differ by only 10%. The 7075-T6 aluminum plates exhibit less deformation than the 6061-T6 plates at the same impact velocity, but at some critical velocity, a through-thickness crack appears in the 7075-T6 plate, ultimately leading to plate perforation. In contrast, the 6061-T6 plates continue to deform and fail by ductile tearing. These differences in damage/failure result in the two alloys having much closer ballistic limit velocities than expected based on differences in strength.

  12. Type VI Secretion System Transports Zn2+ to Combat Multiple Stresses and Host Immunity

    PubMed Central

    Zhu, Wenhan; Gao, Fen; Wang, Yao; Zhang, Lei; Zhang, Weipeng; Wei, Gehong; Luo, Zhao-Qing; Shen, Xihui

    2015-01-01

    Type VI secretion systems (T6SSs) are widespread multi-component machineries that translocate effectors into either eukaryotic or prokaryotic cells, for virulence or for interbacterial competition. Herein, we report that the T6SS-4 from Yersinia pseudotuberculosis displays an unexpected function in the transportation of Zn2+ to combat diverse stresses and host immunity. Environmental insults such as oxidative stress induce the expression of T6SS-4 via OxyR, the transcriptional factor that also regulates many oxidative response genes. Zinc transportation is achieved by T6SS-4-mediated translocation of a novel Zn2+-binding protein substrate YezP (YPK_3549), which has the capacity to rescue the sensitivity to oxidative stress exhibited by T6SS-4 mutants when added to extracellular milieu. Disruption of the classic zinc transporter ZnuABC together with T6SS-4 or yezP results in mutants that almost completely lost virulence against mice, further highlighting the importance of T6SS-4 in resistance to host immunity. These results assigned an unconventional role to T6SSs, which will lay the foundation for studying novel mechanisms of metal ion uptake by bacteria and the role of this process in their resistance to host immunity and survival in harmful environments. PMID:26134274

  13. The corrosion protection of several aluminum alloys by chromic acid and sulfuric acid anodizing

    NASA Technical Reports Server (NTRS)

    Danford, M. D.

    1994-01-01

    The corrosion protection afforded 7075-T6, 7075-T3, 6061-T6, and 2024-T3 aluminum alloys by chromic acid and sulfuric acid anodizing was examined using electrochemical techniques. From these studies, it is concluded that sulfuric acid anodizing provides superior corrosion protection compared to chromic acid anodizing.

  14. Identification of possible virulence marker from Campylobacter jejuni isolates.

    PubMed

    Harrison, James W; Dung, Tran Thi Ngoc; Siddiqui, Fariha; Korbrisate, Sunee; Bukhari, Habib; Tra, My Phan Vu; Hoang, Nguyen Van Minh; Carrique-Mas, Juan; Bryant, Juliet; Campbell, James I; Studholme, David J; Wren, Brendan W; Baker, Stephen; Titball, Richard W; Champion, Olivia L

    2014-06-01

    A novel protein translocation system, the type-6 secretion system (T6SS), may play a role in virulence of Campylobacter jejuni. We investigated 181 C. jejuni isolates from humans, chickens, and environmental sources in Vietnam, Thailand, Pakistan, and the United Kingdom for T6SS. The marker was most prevalent in human and chicken isolates from Vietnam.

  15. Expression of a Yersinia pseudotuberculosis Type VI Secretion System Is Responsive to Envelope Stresses through the OmpR Transcriptional Activator

    PubMed Central

    Gueguen, Erwan; Durand, Eric; Zhang, Xiang Y.; d’Amalric, Quentin; Journet, Laure; Cascales, Eric

    2013-01-01

    The Type VI secretion system (T6SS) is a macromolecular complex widespread in Gram-negative bacteria. Although several T6SS are required for virulence towards host models, most are necessary to eliminate competitor bacteria. Other functions, such as resistance to amoeba predation, biofilm formation or adaptation to environmental conditions have also been reported. This multitude of functions is reflected by the large repertoire of regulatory mechanisms shown to control T6SS expression, production or activation. Here, we demonstrate that one T6SS gene cluster encoded within the Yersinia pseudotuberculosis genome, T6SS-4, is regulated by OmpR, the response regulator of the two-component system EnvZ-OmpR. We first identified OmpR in a transposon mutagenesis screen. OmpR does not control the expression of the four other Y. pseudotuberculosis T6SS gene clusters and of an isolated vgrG gene, and responds to osmotic stresses to bind to and activate the T6SS-4 promoter. Finally, we show that T6SS-4 promotes Y. pseudotuberculosis survival in high osmolarity conditions and resistance to deoxycholate. PMID:23840509

  16. 27 CFR 9.230 - Ballard Canyon.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Los Olivos map, to a marked, unnamed large structure located within a circular-shaped 920-foot contour... located within the 760-foot contour line, T6N/R31W; then (7) Proceed west-southwesterly in a straight line... intersection with the 400-foot contour line, T6N/R31W; then (10) Proceed southerly and then northwesterly...

  17. Independent Regulation of Type VI Secretion in Vibrio cholerae by TfoX and TfoY

    PubMed Central

    Metzger, Lisa C.; Stutzmann, Sandrine; Scrignari, Tiziana; Van der Henst, Charles; Matthey, Noémie; Blokesch, Melanie

    2016-01-01

    Summary Type VI secretion systems (T6SSs) are nanomachines used for interbacterial killing and intoxication of eukaryotes. Although Vibrio cholerae is a model organism for structural studies on T6SSs, the underlying regulatory network is less understood. A recent study showed that the T6SS is part of the natural competence regulon in V. cholerae and is activated by the regulator TfoX. Here, we identify the TfoX homolog TfoY as a second activator of the T6SS. Importantly, despite inducing the same T6SS core machinery, the overall regulons differ significantly for TfoX and TfoY. We show that TfoY does not contribute to competence induction. Instead, TfoY drives the production of T6SS-dependent and T6SS-independent toxins, together with an increased motility phenotype. Hence, we conclude that V. cholerae uses its sole T6SS in response to diverse cues and for distinctive outcomes: either to kill for the prey’s DNA, leading to horizontal gene transfer, or as part of a defensive escape reaction. PMID:27117415

  18. Acid-induced type VI secretion system is regulated by ExoR-ChvG/ChvI signaling cascade in Agrobacterium tumefaciens.

    PubMed

    Wu, Chih-Feng; Lin, Jer-Sheng; Shaw, Gwo-Chyuan; Lai, Erh-Min

    2012-09-01

    The type VI secretion system (T6SS) is a widespread, versatile protein secretion system in pathogenic Proteobacteria. Several T6SSs are tightly regulated by various regulatory systems at multiple levels. However, the signals and/or regulatory mechanisms of many T6SSs remain unexplored. Here, we report on an acid-induced regulatory mechanism activating T6SS in Agrobacterium tumefaciens, a plant pathogenic bacterium causing crown gall disease in a wide range of plants. We monitored the secretion of the T6SS hallmark protein hemolysin-coregulated protein (Hcp) from A. tumefaciens and found that acidity is a T6SS-inducible signal. Expression analysis of the T6SS gene cluster comprising the imp and hcp operons revealed that imp expression and Hcp secretion are barely detected in A. tumefaciens grown in neutral minimal medium but are highly induced with acidic medium. Loss- and gain-of-function analysis revealed that the A. tumefaciens T6SS is positively regulated by a chvG/chvI two-component system and negatively regulated by exoR. Further epistasis analysis revealed that exoR functions upstream of the chvG sensor kinase in regulating T6SS. ChvG protein levels are greatly increased in the exoR deletion mutant and the periplasmic form of overexpressed ExoR is rapidly degraded under acidic conditions. Importantly, ExoR represses ChvG by direct physical interaction, but disruption of the physical interaction allows ChvG to activate T6SS. The phospho-mimic but not wild-type ChvI response regulator can bind to the T6SS promoter region in vitro and activate T6SS with growth in neutral minimal medium. We present the first evidence of T6SS activation by an ExoR-ChvG/ChvI cascade and propose that acidity triggers ExoR degradation, thereby derepressing ChvG/ChvI to activate T6SS in A. tumefaciens. PMID:23028331

  19. Acid-Induced Type VI Secretion System Is Regulated by ExoR-ChvG/ChvI Signaling Cascade in Agrobacterium tumefaciens

    PubMed Central

    Shaw, Gwo-Chyuan; Lai, Erh-Min

    2012-01-01

    The type VI secretion system (T6SS) is a widespread, versatile protein secretion system in pathogenic Proteobacteria. Several T6SSs are tightly regulated by various regulatory systems at multiple levels. However, the signals and/or regulatory mechanisms of many T6SSs remain unexplored. Here, we report on an acid-induced regulatory mechanism activating T6SS in Agrobacterium tumefaciens, a plant pathogenic bacterium causing crown gall disease in a wide range of plants. We monitored the secretion of the T6SS hallmark protein hemolysin-coregulated protein (Hcp) from A. tumefaciens and found that acidity is a T6SS-inducible signal. Expression analysis of the T6SS gene cluster comprising the imp and hcp operons revealed that imp expression and Hcp secretion are barely detected in A. tumefaciens grown in neutral minimal medium but are highly induced with acidic medium. Loss- and gain-of-function analysis revealed that the A. tumefaciens T6SS is positively regulated by a chvG/chvI two-component system and negatively regulated by exoR. Further epistasis analysis revealed that exoR functions upstream of the chvG sensor kinase in regulating T6SS. ChvG protein levels are greatly increased in the exoR deletion mutant and the periplasmic form of overexpressed ExoR is rapidly degraded under acidic conditions. Importantly, ExoR represses ChvG by direct physical interaction, but disruption of the physical interaction allows ChvG to activate T6SS. The phospho-mimic but not wild-type ChvI response regulator can bind to the T6SS promoter region in vitro and activate T6SS with growth in neutral minimal medium. We present the first evidence of T6SS activation by an ExoR-ChvG/ChvI cascade and propose that acidity triggers ExoR degradation, thereby derepressing ChvG/ChvI to activate T6SS in A. tumefaciens. PMID:23028331

  20. Structure and Regulation of the Type VI Secretion System

    PubMed Central

    Silverman, Julie M.; Brunet, Yannick R.; Cascales, Eric; Mougous, Joseph D.

    2013-01-01

    The type VI secretion system (T6SS) is a complex and widespread gram-negative export pathway with the capacity to translocate protein effectors into a diversity of target cell types. Current structural models of the T6SS indicate that the apparatus is composed of at least two complexes, a dynamic bacteriophage-like structure and a cell envelope-spanning membrane-associated assembly. How these complexes interact to promote effector secretion and cell targeting remains a major question in the field. As a contact-dependent pathway with specific cellular targets, the T6SS is subject to tight regulation. Thus, the identification of regulatory elements that control T6S expression continues to shape our understanding of the environmental circumstances relevant to its function. This review discusses recent progress toward characterizing T6S structure and regulation. PMID:22746332

  1. The Type VI Secretion System Encoded in Salmonella Pathogenicity Island 19 Is Required for Salmonella enterica Serotype Gallinarum Survival within Infected Macrophages

    PubMed Central

    Blondel, Carlos J.; Jiménez, Juan C.; Leiva, Lorenzo E.; Álvarez, Sergio A.; Pinto, Bernardo I.; Contreras, Francisca; Pezoa, David; Santiviago, Carlos A.

    2013-01-01

    Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells. PMID:23357385

  2. The type VI secretion system encoded in Salmonella pathogenicity island 19 is required for Salmonella enterica serotype Gallinarum survival within infected macrophages.

    PubMed

    Blondel, Carlos J; Jiménez, Juan C; Leiva, Lorenzo E; Alvarez, Sergio A; Pinto, Bernardo I; Contreras, Francisca; Pezoa, David; Santiviago, Carlos A; Contreras, Inés

    2013-04-01

    Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells.

  3. The Type VI Secretion System Modulates Flagellar Gene Expression and Secretion in Citrobacter freundii and Contributes to Adhesion and Cytotoxicity to Host Cells.

    PubMed

    Liu, Liyun; Hao, Shuai; Lan, Ruiting; Wang, Guangxia; Xiao, Di; Sun, Hui; Xu, Jianguo

    2015-07-01

    The type VI secretion system (T6SS) as a virulence factor-releasing system contributes to virulence development of various pathogens and is often activated upon contact with target cells. Citrobacter freundii strain CF74 has a complete T6SS genomic island (GI) that contains clpV, hcp-2, and vgr T6SS genes. We constructed clpV, hcp-2, vgr, and T6SS GI deletion mutants in CF74 and analyzed their effects on the transcriptome overall and, specifically, on the flagellar system at the levels of transcription and translation. Deletion of the T6SS GI affected the transcription of 84 genes, with 15 and 69 genes exhibiting higher and lower levels of transcription, respectively. Members of the cell motility class of downregulated genes of the CF74ΔT6SS mutant were mainly flagellar genes, including effector proteins, chaperones, and regulators. Moreover, the production and secretion of FliC were also decreased in clpV, hcp-2, vgr, or T6SS GI deletion mutants in CF74 and were restored upon complementation. In swimming motility assays, the mutant strains were found to be less motile than the wild type, and motility was restored by complementation. The mutant strains were defective in adhesion to HEp-2 cells and were restored partially upon complementation. Further, the CF74ΔT6SS, CF74ΔclpV, and CF74Δhcp-2 mutants induced lower cytotoxicity to HEp-2 cells than the wild type. These results suggested that the T6SS GI in CF74 regulates the flagellar system, enhances motility, is involved in adherence to host cells, and induces cytotoxicity to host cells. Thus, the T6SS plays a wide-ranging role in C. freundii.

  4. Wheat Grain Development Is Characterized by Remarkable Trehalose 6-Phosphate Accumulation Pregrain Filling: Tissue Distribution and Relationship to SNF1-Related Protein Kinase1 Activity1[W][OA

    PubMed Central

    Martínez-Barajas, Eleazar; Delatte, Thierry; Schluepmann, Henriette; de Jong, Gerhardus J.; Somsen, Govert W.; Nunes, Cátia; Primavesi, Lucia F.; Coello, Patricia; Mitchell, Rowan A.C.; Paul, Matthew J.

    2011-01-01

    Trehalose 6-phosphate (T6P) is a sugar signal that regulates metabolism, growth, and development and inhibits the central regulatory SNF1-related protein kinase1 (SnRK1; AKIN10/AKIN11). To better understand the mechanism in wheat (Triticum aestivum) grain, we analyze T6P content and SnRK1 activities. T6P levels changed 178-fold 1 to 45 d after anthesis (DAA), correlating with sucrose content. T6P ranged from 78 nmol g−1 fresh weight (FW) pregrain filling, around 100-fold higher than previously reported in plants, to 0.4 nmol g−1 FW during the desiccation stage. In contrast, maximum SnRK1 activity changed only 3-fold but was inhibited strongly by T6P in vitro. To assess SnRK1 activity in vivo, homologs of SnRK1 marker genes in the wheat transcriptome were identified using Wheat Estimated Transcript Server. SnRK1-induced and -repressed marker genes were expressed differently pregrain filling compared to grain filling consistent with changes in T6P. To investigate this further maternal and filial tissues were compared pre- (7 DAA) and during grain filling (17 DAA). Strikingly, in vitro SnRK1 activity was similar in all tissues in contrast to large changes in tissue distribution of T6P. At 7 DAA T6P was 49 to 119 nmol g−1 FW in filial and maternal tissues sufficient to inhibit SnRK1; at 17 DAA T6P accumulation was almost exclusively endospermal (43 nmol g−1 FW) with 0.6 to 0.8 nmol T6P g−1 FW in embryo and pericarp. The data show a correlation between T6P and sucrose overall that belies a marked effect of tissue type and developmental stage on T6P content, consistent with tissue-specific regulation of SnRK1 by T6P in wheat grain. PMID:21402798

  5. Estudio del comportamiento tribologico y de las interacciones de superficie de nuevos nanofluidos ionicos

    NASA Astrophysics Data System (ADS)

    Espinosa Rodriguez, Tulia

    tribocorrosion processes. The formation of a coating layer on magnesium alloys from phosphonate imidazolium ionic liquids by immersion and by chronoamperometry has been described. The new coatings reduce the abrasive wear in the magnesium-aluminium alloy but they are not effective in the magnesium-zinc alloy, which prevent the formation of continuous coatings. Los liquidos ionicos son sales liquidas a temperatura ambiente o bajas temperaturas que presentan excelentes propiedades fisico-quimicas. En el presente trabajo se estudian como lubricantes en problemas tribologicos complejos como la lubricacion de metales contra si mismos, el desarrollo de lubricantes base agua y de nuevas superficies autolubricadas. Cuando no es posible reducir la friccion y desgaste mediante lubricacion, como en las aleaciones de magnesio, los liquidos ionicos se han estudiado como precursores de recubrimientos protectores. Se han determinado las interacciones superficiales y los procesos de corrosion sobre cobre y sobre acero con diferentes liquidos ionicos proticos y aproticos para desarrollar nuevos lubricantes y aditivos. En el contacto cobre/cobre, excepto el liquido ionico protico derivado del oleato, todos los liquidos ionicos estudiados presentan mejor comportamiento tribologico que el lubricante comercial Polialfaolefina 6. En el contacto acero/zafiro, los nuevos liquidos ionicos proticos son buenos lubricantes cuando se utilizan en estado puro, y, como aditivos en agua, generan peliculas adsorbidas sobre la superficie del metal reduciendo la friccion y el desgaste tras la evaporacion del agua. Para evitar el periodo de alta friccion inicial en presencia de agua, se han generado peliculas superficiales de liquido ionico sobre el acero en condiciones estaticas. El mejor comportamiento lubricante tanto en el contacto cobre/cobre como en el contacto acero/zafiro se obtiene para el liquido ionico protico derivado del anion adipato, con dos grupos carboxilicos. Las interacciones de los grupos

  6. A view to a kill: the bacterial type 6 secretion system

    PubMed Central

    Ho, Brian T.; Dong, Tao G.; Mekalanos, John J.

    2014-01-01

    The bacterial Type 6 Secretion System (T6SS) is an organelle that is structurally and mechanistically analogous to an intracellular membrane-attached contractile phage tail. Recent studies determined that a rapid conformational change in the structure of a sheath protein complex propels T6SS spike and tube components along with anti-bacterial and anti-eukaryotic effectors out of predatory T6SS+ cells and into prey cells. The contracted organelle is then recycled in an ATP-dependent process. T6SS is regulated at transcriptional and post-translational levels, the latter involving detection of membrane perturbation in some species. In addition to directly targeting eukaryotic cells, the T6SS can also target other bacteria co-infecting a mammalian host, highlighting the importance of the T6SS not only for bacterial survival in environmental ecosystems but also in the context of infection and disease. This review highlights these and other advances in our understanding of the structure, mechanical function, assembly, and regulation of the T6SS. PMID:24332978

  7. How do sugars regulate plant growth and development? New insight into the role of trehalose-6-phosphate.

    PubMed

    O'Hara, Liam E; Paul, Matthew J; Wingler, Astrid

    2013-03-01

    Plant growth and development are tightly controlled in response to environmental conditions that influence the availability of photosynthetic carbon in the form of sucrose. Trehalose-6-phosphate (T6P), the precursor of trehalose in the biosynthetic pathway, is an important signaling metabolite that is involved in the regulation of plant growth and development in response to carbon availability. In addition to the plant's own pathway for trehalose synthesis, formation of T6P or trehalose by pathogens can result in the reprogramming of plant metabolism and development. Developmental processes that are regulated by T6P range from embryo development to leaf senescence. Some of these processes are regulated in interaction with phytohormones, such as auxin. A key interacting factor of T6P signaling in response to the environment is the protein kinase sucrose non-fermenting related kinase-1 (SnRK1), whose catalytic activity is inhibited by T6P. SnRK1 is most likely involved in the adjustment of metabolism and growth in response to starvation. The transcription factor bZIP11 has recently been identified as a new player in the T6P/SnRK1 regulatory pathway. By inhibiting SnRK1, T6P promotes biosynthetic reactions. This regulation has important consequences for crop production, for example, in the developing wheat grain and during the growth of potato tubers.

  8. Identification of divergent type VI secretion effectors using a conserved chaperone domain

    PubMed Central

    Liang, Xiaoye; Moore, Richard; Wilton, Mike; Wong, Megan J. Q.; Lam, Linh; Dong, Tao G.

    2015-01-01

    The type VI secretion system (T6SS) is a lethal weapon used by many bacteria to kill eukaryotic predators or prokaryotic competitors. Killing by the T6SS results from repetitive delivery of toxic effectors. Despite their importance in dictating bacterial fitness, systematic prediction of T6SS effectors remains challenging due to high effector diversity and the absence of a conserved signature sequence. Here, we report a class of T6SS effector chaperone (TEC) proteins that are required for effector delivery through binding to VgrG and effector proteins. The TEC proteins share a highly conserved domain (DUF4123) and are genetically encoded upstream of their cognate effector genes. Using the conserved TEC domain sequence, we identified a large family of TEC genes coupled to putative T6SS effectors in Gram-negative bacteria. We validated this approach by verifying a predicted effector TseC in Aeromonas hydrophila. We show that TseC is a T6SS-secreted antibacterial effector and that the downstream gene tsiC encodes the cognate immunity protein. Further, we demonstrate that TseC secretion requires its cognate TEC protein and an associated VgrG protein. Distinct from previous effector-dependent bioinformatic analyses, our approach using the conserved TEC domain will facilitate the discovery and functional characterization of new T6SS effectors in Gram-negative bacteria. PMID:26150500

  9. Chimeric adaptor proteins translocate diverse type VI secretion system effectors in Vibrio cholerae.

    PubMed

    Unterweger, Daniel; Kostiuk, Benjamin; Ötjengerdes, Rina; Wilton, Ashley; Diaz-Satizabal, Laura; Pukatzki, Stefan

    2015-08-13

    Vibrio cholerae is a diverse species of Gram-negative bacteria, commonly found in the aquatic environment and the causative agent of the potentially deadly disease cholera. These bacteria employ a type VI secretion system (T6SS) when they encounter prokaryotic and eukaryotic competitors. This contractile puncturing device translocates a set of effector proteins into neighboring cells. Translocated effectors are toxic unless the targeted cell produces immunity proteins that bind and deactivate incoming effectors. Comparison of multiple V. cholerae strains indicates that effectors are encoded in T6SS effector modules on mobile genetic elements. We identified a diverse group of chimeric T6SS adaptor proteins required for the translocation of diverse effectors encoded in modules. An example for a T6SS effector that requires T6SS adaptor protein 1 (Tap-1) is TseL found in pandemic V. cholerae O1 serogroup strains and other clinical isolates. We propose a model in which Tap-1 is required for loading TseL onto the secretion apparatus. After T6SS-mediated TseL export is completed, Tap-1 is retained in the bacterial cell to load other T6SS machines.

  10. Structure/property relations of aluminum under varying rates and stress states

    SciTech Connect

    Tucker, Matthew T; Horstemeyer, Mark F; Whittington, Wilburn R; Solanki, Kiran N

    2010-11-19

    In this work we analyze the plasticity, damage, and fracture characteristics of three different processed aluminum alloys (rolled 5083-H13, cast A356-T6, and extruded 6061-T6) under varying stress states (tension, compression, and torsion) and strain rates (0.001/, 1/s., and 1000/s). The stress state difference had more of a flow stress effect than the applied strain rates for those given in this study (0.001/sec up to 1000/sec). The stress state and strain rate also had a profound effect on the damage evolution of each aluminum alloy. Tension and torsional straining gave much greater damage nucleation rates than compression. Although the damage of all three alloys was found to be void nucleation dominated, the A356-T6 and 5083-H131 aluminum alloys incurred void damage via micron scale particles where the 6061-T6 aluminum alloy incurred void damage from two scales, micron-scale particles and nanoscale precipitates. Having two length scales of particles that participated in the damage evolution made the 6061-T6 incur a strain rate sensitive damage rate that was different than the other two aluminum alloys. Under tension, as the strain rate increased, the 6061-T6 aluminum alloy's void nucleation rate decreased, but the A356-T6 and 5083-H131 aluminum alloys void nucleation rate increased.

  11. Tensile and compressive stress-strain behavior of heat treated boron-aluminum

    NASA Technical Reports Server (NTRS)

    Kennedy, J. M.; Tenney, D. R.; Herakovich, C. T.

    1978-01-01

    An experimental study was conducted to assess the effects of heat treatment and cyclic mechanical loading on the tensile and compressive stress-strain behavior of six boron-aluminum composites having different laminate orientations and being subjected to different heat treatments. The heat treatments were as-fabricated, T6, and T6N consisting of T6 treatment followed by cryogenic quench in liquid nitrogen prior to testing. All laminates were tested in monotonic and cyclic compression, while the tensile-test data are taken from the literature for comparison purposes. It is shown that the linear elastic range of the T6- and T6N-condition specimens is larger than that of the as-fabricated specimens, and that cyclic loading in tension or compression strain hardens the specimens and extends the linear elastic range. For laminates containing 0-deg plies, the stress-strain behavior upon unloading is found to be nonlinear, whereas the other laminates exhibit a linear behavior upon unloading. Specimens in the T6 and T6N conditions show higher strain hardening than the as-fabricated specimens.

  12. Type VI Secretion System Toxins Horizontally Shared between Marine Bacteria

    PubMed Central

    Salomon, Dor; Klimko, John A.; Trudgian, David C.; Kinch, Lisa N.; Grishin, Nick V.; Mirzaei, Hamid; Orth, Kim

    2015-01-01

    The type VI secretion system (T6SS) is a widespread protein secretion apparatus used by Gram-negative bacteria to deliver toxic effector proteins into adjacent bacterial or host cells. Here, we uncovered a role in interbacterial competition for the two T6SSs encoded by the marine pathogen Vibrio alginolyticus. Using comparative proteomics and genetics, we identified their effector repertoires. In addition to the previously described effector V12G01_02265, we identified three new effectors secreted by T6SS1, indicating that the T6SS1 secretes at least four antibacterial effectors, of which three are members of the MIX-effector class. We also showed that the T6SS2 secretes at least three antibacterial effectors. Our findings revealed that many MIX-effectors belonging to clan V are “orphan” effectors that neighbor mobile elements and are shared between marine bacteria via horizontal gene transfer. We demonstrated that a MIX V-effector from V. alginolyticus is a functional T6SS effector when ectopically expressed in another Vibrio species. We propose that mobile MIX V-effectors serve as an environmental reservoir of T6SS effectors that are shared and used to diversify antibacterial toxin repertoires in marine bacteria, resulting in enhanced competitive fitness. PMID:26305100

  13. An ERP Study of Good Production vis-à-vis Poor Perception of Tones in Cantonese: Implications for Top-Down Speech Processing

    PubMed Central

    Law, Sam-Po; Fung, Roxana; Kung, Carmen

    2013-01-01

    This study investigated a theoretically challenging dissociation between good production and poor perception of tones among neurologically unimpaired native speakers of Cantonese. The dissociation is referred to as the near-merger phenomenon in sociolinguistic studies of sound change. In a passive oddball paradigm, lexical and nonlexical syllables of the T1/T6 and T4/T6 contrasts were presented to elicit the mismatch negativity (MMN) and P3a from two groups of participants, those who could produce and distinguish all tones in the language (Control) and those who could produce all tones but specifically failed to distinguish between T4 and T6 in perception (Dissociation). The presence of MMN to T1/T6 and null response to T4/T6 of lexical syllables in the dissociation group confirmed the near-merger phenomenon. The observation that the control participants exhibited a statistically reliable MMN to lexical syllables of T1/T6, weaker responses to nonlexical syllables of T1/T6 and lexical syllables of T4/T6, and finally null response to nonlexical syllables of T4/T6, suggests the involvement of top-down processing in speech perception. Furthermore, the stronger P3a response of the control group, compared with the dissociation group in the same experimental conditions, may be taken to indicate higher cognitive capability in attention switching, auditory attention or memory in the control participants. This cognitive difference, together with our speculation that constant top-down predictions without complete bottom-up analysis of acoustic signals in speech recognition may reduce one’s sensitivity to small acoustic contrasts, account for the occurrence of dissociation in some individuals but not others. PMID:23342146

  14. An ERP study of good production vis-à-vis poor perception of tones in Cantonese: implications for top-down speech processing.

    PubMed

    Law, Sam-Po; Fung, Roxana; Kung, Carmen

    2013-01-01

    This study investigated a theoretically challenging dissociation between good production and poor perception of tones among neurologically unimpaired native speakers of Cantonese. The dissociation is referred to as the near-merger phenomenon in sociolinguistic studies of sound change. In a passive oddball paradigm, lexical and nonlexical syllables of the T1/T6 and T4/T6 contrasts were presented to elicit the mismatch negativity (MMN) and P3a from two groups of participants, those who could produce and distinguish all tones in the language (Control) and those who could produce all tones but specifically failed to distinguish between T4 and T6 in perception (Dissociation). The presence of MMN to T1/T6 and null response to T4/T6 of lexical syllables in the dissociation group confirmed the near-merger phenomenon. The observation that the control participants exhibited a statistically reliable MMN to lexical syllables of T1/T6, weaker responses to nonlexical syllables of T1/T6 and lexical syllables of T4/T6, and finally null response to nonlexical syllables of T4/T6, suggests the involvement of top-down processing in speech perception. Furthermore, the stronger P3a response of the control group, compared with the dissociation group in the same experimental conditions, may be taken to indicate higher cognitive capability in attention switching, auditory attention or memory in the control participants. This cognitive difference, together with our speculation that constant top-down predictions without complete bottom-up analysis of acoustic signals in speech recognition may reduce one's sensitivity to small acoustic contrasts, account for the occurrence of dissociation in some individuals but not others. PMID:23342146

  15. Oceanobacillus-like bacterium isolated from Vyhna travertine spring.

    PubMed

    Pristas, P; Cunderlikova, M; Judova, J

    2014-03-01

    During characterization of autochthonic Vyhna travertine source microflora, several bacterial strains were isolated and characterised. Isolate T6, a halotolerant, moderately alkaliphilic and thermophilic bacterial isolate, was further characterised based on physiological, microbiological and biochemical tests and phylogenetic 16S rRNA analysis. On the basis of the results obtained, the T6 isolate should be placed in the genus Oceanobacillus, and it is probably a prototype of a novel bacterial species. Characterization of the T6 isolate broadens our knowledge on variability of halophilic bacteria of Oceanobacillus genus and expands data on travertine-associated bacterial communities. PMID:24022266

  16. Human amniotic fluid for fertilization and culture of human embryos: results of clinical trials in human in vitro fertilization (IVF) programs.

    PubMed

    Gianaroli, L; Trounson, A; King, C; Ferraretti, A; Chiappazzo, L; Bafaro, G

    1989-08-01

    We report the outcome of clinical trials carried out in two IVF programs, comparing the use of human amniotic fluid (HAF) as a complete medium to Whittingham's T6 medium containing human serum (T6 + 10% HS) for egg incubation, insemination, embryo culture, and embryo transfer. There were no significant differences in the clinical trials between HAF used alone as a complete medium and T6 + 10% HS in fertilization rates of eggs, cleavage rates of embryos up to 48 hours in culture, pregnancy success rates after embryo replacement or the outcome of pregnancies. There was no advantage in using T6 + 10% HS for fertilization of eggs and HAF as a complete medium for embryo culture and transfer in any of the parameters examined. We conclude that HAF does not meet the complete requirements of human eggs and embryos in vitro and further developments of culture media are required to obtain embryo development equivalent to that in vivo.

  17. Crystal structure of YwpF from Staphylococcus aureus reveals its architecture comprised of a β-barrel core domain resembling type VI secretion system proteins and a two-helix pair.

    PubMed

    Lee, Sang Jae; Lee, Kyu-Yeon; Lee, Ki-Young; Kim, Dong-Gyun; Kim, Soon-Jong; Lee, Bong-Jin

    2015-04-01

    The ywpF gene (SAV2097) of the Staphylococcus aureus strain Mu50 encodes the YwpF protein, which may play a role in antibiotic resistance. Here, we report the first crystal structure of the YwpF superfamily from S. aureus at 2.5-Å resolution. The YwpF structure consists of two regions: an N-terminal core β-barrel domain that shows structural similarity to type VI secretion system (T6SS) proteins (e.g., Hcp1, Hcp3, and EvpC) and a C-terminal two-helix pair. Although the monomer structure of S. aureus YwpF resembles those of T6SS proteins, the dimer/tetramer model of S. aureus YwpF is distinct from the functionally important hexameric ring of T6SS proteins. We therefore suggest that the S. aureus YwpF may have a different function compared to T6SS proteins.

  18. Aim, Load, Fire: The Type VI Secretion System, a Bacterial Nanoweapon.

    PubMed

    Cianfanelli, Francesca R; Monlezun, Laura; Coulthurst, Sarah J

    2016-01-01

    Bacteria utilise specialised protein secretion systems to interact with host organisms, competitor bacteria, and the environment. The Type VI secretion system (T6SS) is a versatile weapon deployed by many bacterial species to target either host cells or rival bacteria. The widespread occurrence and significance of the T6SS is becoming increasingly appreciated, as is its intriguing mode of action. The T6SS delivers multiple, diverse effector proteins directly into target cells using a dynamic 'firing' mechanism related to the action of contractile bacteriophage tails. Here, we summarise the contribution of recent findings to our developing picture of how the T6SS assembles and fires, how it is loaded with different types of effectors, and how it can be aimed towards an incoming assault. PMID:26549582

  19. Stress-corrosion-induced property changes in aluminum alloys

    NASA Technical Reports Server (NTRS)

    Bankston, B. F.; Clotfelter, W. N.

    1968-01-01

    Measurements of electrical conductivity, ultrasonic surface wave attenuation, and internal friction loss were made on aluminum alloys 7079-T6, 2219-T31, and 2219-T81 as a function of the onset of stress corrosion.

  20. [Histiocytosis X. An oligosymptomatic form of Hand-Schüller-Christian disease with a new form and possibilities of diagnosis].

    PubMed

    Eberth-Willershausen, W; Steger, O; Barran, W; Burg, G; Ryckmanns, F; Kaudewitz, P

    1984-10-01

    A patient is reported who had a monosymptomatic, cutaneous form of histiocytosis X. The reactivity of cutaneous histiocytosis-X cells with monoclonal anti-T-6 antibody is discussed as a new diagnostic procedure.

  1. Source/sink interactions underpin crop yield: the case for trehalose 6-phosphate/SnRK1 in improvement of wheat.

    PubMed

    Lawlor, David W; Paul, Matthew J

    2014-01-01

    Considerable interest has been evoked by the analysis of the regulatory pathway in carbohydrate metabolism and cell growth involving the non-reducing disaccharide trehalose (TRE). TRE is at small concentrations in mesophytes such as Arabidopsis thaliana and Triticum aestivum, excluding a role in osmoregulation once suggested for it. Studies of TRE metabolism, and genetic modification of it, have shown a very wide and more important role of the pathway in regulation of many processes in development, growth, and photosynthesis. It has now been established that rather than TRE, it is trehalose 6-phosphate (T6P) which has such profound effects. T6P is the intermediary in TRE synthesis formed from glucose-6-phosphate and UDP-glucose, derived from sucrose, by the action of trehalose phosphate synthase. The concentration of T6P is determined both by the rate of synthesis, which depends on the sucrose concentration, and also by the rate of breakdown by trehalose-6-phosphate phosphatase which produces TRE. Changing T6P concentrations by genetically modifying the enzymes of synthesis and breakdown has altered photosynthesis, sugar metabolism, growth, and development which affect responses to, and recovery from, environmental factors. Many of the effects of T6P on metabolism and growth occur via the interaction of T6P with the SnRK1 protein kinase system. T6P inhibits the activity of SnRK1, which de-represses genes encoding proteins involved in anabolism. Consequently, a large concentration of sucrose increases T6P and thereby inhibits SnRK1, so stimulating growth of cells and their metabolic activity. The T6P/SnRK1 mechanism offers an important new view of how the distribution of assimilates to organs, such as developing grains in cereal plants, is achieved. This review briefly summarizes the factors determining, and limiting, yield of wheat (particularly mass/grain which is highly conserved) and considers how T6P/SnRK1 might function to determine grain yield and might be

  2. Evaluation of the roles played by Hcp and VgrG type 6 secretion system effectors in Aeromonas hydrophila SSU pathogenesis

    PubMed Central

    Sha, Jian; Rosenzweig, Jason A.; Kozlova, Elena V.; Wang, Shaofei; Erova, Tatiana E.; Kirtley, Michelle L.; van Lier, Christina J.

    2013-01-01

    Aeromonas hydrophila, a Gram-negative bacterium, is an emerging human pathogen equipped with both a type 3 and a type 6 secretion system (T6SS). In this study, we evaluated the roles played by paralogous T6SS effector proteins, hemolysin co-regulated proteins (Hcp-1 and -2) and valine glycine repeat G (VgrG-1, -2 and -3) protein family members in A. hydrophila SSU pathogenesis by generating various combinations of deletion mutants of the their genes. In addition to their predicted roles as structural components and effector proteins of the T6SS, our data clearly demonstrated that paralogues of Hcp and VgrG also influenced bacterial motility, protease production and biofilm formation. Surprisingly, there was limited to no observed functional redundancy among and/or between the aforementioned T6SS effector paralogues in multiple assays. Our data indicated that Hcp and VgrG paralogues located within the T6SS cluster were more involved in forming T6SS structures, while the primary roles of Hcp-1 and VgrG-1, located outside of the T6SS cluster, were as T6SS effectors. In terms of influence on bacterial physiology, Hcp-1, but not Hcp-2, influenced bacterial motility and protease production, and in its absence, increases in both of the aforementioned activities were observed. Likewise, VgrG-1 played a major role in regulating bacterial protease production, while VgrG-2 and VgrG-3 were critical in regulating bacterial motility and biofilm formation. In an intraperitoneal murine model of infection, all Hcp and VgrG paralogues were required for optimal bacterial virulence and dissemination to mouse peripheral organs. Importantly, the observed phenotypic alterations of the T6SS mutants could be fully complemented. Taking these results together, we have further established the roles played by the two known T6SS effectors of A. hydrophila by defining their contributions to T6SS function and virulence in both in vitro and in vivo models of infection. PMID:23519162

  3. Trehalose 6-phosphate regulates starch synthesis via posttranslational redox activation of ADP-glucose pyrophosphorylase.

    PubMed

    Kolbe, Anna; Tiessen, Axel; Schluepmann, Henriette; Paul, Matthew; Ulrich, Silke; Geigenberger, Peter

    2005-08-01

    Trehalose is the most widespread disaccharide in nature, occurring in bacteria, fungi, insects, and plants. Its precursor, trehalose 6-phosphate (T6P), is also indispensable for the regulation of sugar utilization and growth, but the sites of action are largely unresolved. Here we use genetic and biochemical approaches to investigate whether T6P acts to regulate starch synthesis in plastids of higher plants. Feeding of trehalose to Arabidopsis leaves led to stimulation of starch synthesis within 30 min, accompanied by activation of ADP-glucose pyrophosphorylase (AGPase) via posttranslational redox modification. The response resembled sucrose but not glucose feeding and depended on the expression of SNF1-related kinase. We also analyzed transgenic Arabidopsis plants with T6P levels increased by expression of T6P synthase or decreased by expression of T6P phosphatase (TPP) in the cytosol. Compared with wild type, leaves of T6P synthase-expressing plants had increased redox activation of AGPase and increased starch, whereas TPP-expressing plants showed the opposite. Moreover, TPP expression prevented the increase in AGPase activation in response to sucrose or trehalose feeding. Incubation of intact isolated chloroplasts with 100 muM T6P significantly and specifically increased reductive activation of AGPase within 15 min. Results provide evidence that T6P is synthesized in the cytosol and acts on plastidial metabolism by promoting thioredoxin-mediated redox transfer to AGPase in response to cytosolic sugar levels, thereby allowing starch synthesis to be regulated independently of light. The discovery informs about the evolution of plant metabolism and how chloroplasts of prokaryotic origin use an intermediate of the ancient trehalose pathway to report the metabolic status of the cytosol.

  4. Fatigue Performance of Fluidized Bed Heat Treated 319 Alloy Diesel Cylinder Heads

    NASA Astrophysics Data System (ADS)

    Chaudhury, Sujoy K.; Apelian, Diran; Meyer, Philippe; Massinon, Denis; Morichon, Julien

    2015-07-01

    Effects of various heat treatment tempers on fatigue performance of 319 alloy diesel cylinder heads were investigated. Castings were heat treated to T5, T6, and T7 tempers. Castings were solution heat treated and quenched using fluidized beds and aged using both conventional air convective furnace and fluidized bed for T6 and T7 tempers; while they were aged after casting for T5 temper using conventional furnace. Fatigue tests were performed at 373 K (100 °C) and stress ratio equal to -1. Results show that heat treatment has significant effect on the fatigue behavior of 319 alloy. The fatigue strength of T6 tempered 319 alloy is greater than T5 and T7 treatments. Weibull analysis shows that the Weibull modulus and characteristic fatigue life of castings treated (using conventional forced air circulation electrical resistance furnace) to T6 and T7 tempers are greater than T5 temper. This implies that castings treated to T6 and T7 tempers have greater reliability vis-à-vis T5 temper. Fractographic analyses reveal three distinct regions. These are: (I) crack initiation region from the surface, (II) crack propagation region, and (III) catastrophic or monotonic failure region. The relative size of the crack propagation region in T6 and T7 treated samples is greater than T5 treated samples. In general, the monotonic failure region shows typical dimple morphology, which implies significant plastic deformation prior to failure. Dimples on the fractured surface of T5 treated alloy are relatively more faceted than those treated to T6 and T7 tempers. This implies that the 319 alloy treated to T6 and T7 tempers underwent higher degree of plasticity prior to failure than that in the T5 condition.

  5. Quorum sensing and a global regulator TsrA control expression of type VI secretion and virulence in Vibrio cholerae.

    PubMed

    Zheng, Jun; Shin, Ok S; Cameron, D Ewen; Mekalanos, John J

    2010-12-01

    Vibrio cholerae is a human pathogen that causes the life-threatening diarrheal disease cholera. A type VI secretion system (T6SS) was recently shown to be required for full virulence in the O37 serogroup strain V52, which causes only sporadic human disease, but T6SS is not expressed in seventh pandemic O1 El Tor strains under standard laboratory conditions. In this study, we show that in the O1 El Tor strain C6706, T6SS is repressed by both quorum sensing and the uncharacterized protein VC0070 (TsrA). Disruption of TsrA and the quorum sensing regulator LuxO induces expression and secretion of the T6SS substrate Hcp, and this is dependent on the downstream regulator HapR, which directly binds to the promoter region of the T6SS genes hcp1 and hcp2 to induce expression. The activated T6SS in C6706 is functional and can translocate the effector protein VgrG-1 into macrophage cells, and T6SS activation leads to fecal diarrhea and intestinal inflammation in infant rabbits. Using an infant mouse infection model, we show that deletion of tsrA results in a 9.3-fold increase in intestinal colonization compared with wild type. TsrA functions as a global regulator to activate expression of hemagglutinin protease and repress cholera toxin and toxin coregulated pilus. Our findings provide significant insight into the molecular mechanism of T6SS and ToxT regulon gene regulation by quorum sensing and TsrA.

  6. Genetically distinct pathways guide effector export through the type VI secretion system

    PubMed Central

    Whitney, John C.; Beck, Christina M.; Goo, Young Ah; Russell, Alistair B.; Harding, Brittany; De Leon, Justin A.; Cunningham, David A.; Tran, Bao Q.; Low, David A.; Goodlett, David R.; Hayes, Christopher S.; Mougous, Joseph D.

    2014-01-01

    Summary Bacterial secretion systems often employ molecular chaperones to recognize and facilitate export of their substrates. Recent work demonstrated that a secreted component of the type VI secretion system (T6SS), hemolysin co-regulated protein (Hcp), binds directly to effectors, enhancing their stability in the bacterial cytoplasm. Herein, we describe a quantitative cellular proteomics screen for T6S substrates that exploits this chaperone-like quality of Hcp. Application of this approach to the Hcp secretion island I-encoded T6SS (H1-T6SS) of Pseudomonas aeruginosa led to the identification of a novel effector protein, termed Tse4 (type VI secretion exported 4), subsequently shown to act as a potent intra-specific H1-T6SS-delivered antibacterial toxin. Interestingly, our screen failed to identify two predicted H1-T6SS effectors, Tse5 and Tse6, which differ from Hcp-stabilized substrates by the presence of toxin-associated PAAR-repeat motifs and genetic linkage to members of the valine-glycine repeat protein G (vgrG) genes. Genetic studies further distinguished these two groups of effectors: Hcp-stabilized effectors were found to display redundancy in interbacterial competition with respect to the requirement for the two H1-T6SS-exported VgrG proteins, whereas Tse5 and Tse6 delivery strictly required a cognate VgrG. Together, we propose that interaction with either VgrG or Hcp defines distinct pathways for T6S effector export. PMID:24589350

  7. 40 CFR 81.313 - Idaho.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    .... Editorial Note: For Federal Register citations affecting § 81.313 see the List of CFR Sections Affected... Area: T.5S, R.34E Sections 25-36 T.5S, R.35E Section 31 T.6S, R.34E Sections 1-36 T.6S, R.35E Sections.../90 Nonattainment 11/15/90 Moderate Fort Hall Indian Reservation: T.5S, R.34E Sections 15-23 T.5S,...

  8. 27 CFR 9.162 - Sta. Rita Hills.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ..., elevation 1600 feet, in section 27, T.6N, R. 32W, on the Solvang, Calif., Quadrangle U.S.G.S. map. (2) Then proceed north and slightly west 2.3 miles to an unnamed hilltop elevation 1174 feet, Section 15, T.6N., R. 32W. (3) Proceed west and slightly north 1.85 miles to an unnamed hilltop elevation 899 feet...

  9. 27 CFR 9.162 - Sta. Rita Hills.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., elevation 1600 feet, in section 27, T.6N, R. 32W, on the Solvang, Calif., Quadrangle U.S.G.S. map. (2) Then proceed north and slightly west 2.3 miles to an unnamed hilltop elevation 1174 feet, Section 15, T.6N., R. 32W. (3) Proceed west and slightly north 1.85 miles to an unnamed hilltop elevation 899 feet...

  10. Gene Expression of Type VI Secretion System Associated with Environmental Survival in Acidovorax avenae subsp. avenae by Principle Component Analysis.

    PubMed

    Cui, Zhouqi; Jin, Guoqiang; Li, Bin; Kakar, Kaleem Ullah; Ojaghian, Mohammad Reza; Wang, Yangli; Xie, Guanlin; Sun, Guochang

    2015-09-11

    Valine glycine repeat G (VgrG) proteins are regarded as one of two effectors of Type VI secretion system (T6SS) which is a complex multi-component secretion system. In this study, potential biological roles of T6SS structural and VgrG genes in a rice bacterial pathogen, Acidovorax avenae subsp. avenae (Aaa) RS-1, were evaluated under seven stress conditions using principle component analysis of gene expression. The results showed that growth of the pathogen was reduced by H₂O₂ and paraquat-induced oxidative stress, high salt, low temperature, and vgrG mutation, compared to the control. However, pathogen growth was unaffected by co-culture with a rice rhizobacterium Burkholderia seminalis R456. In addition, expression of 14 T6SS structural and eight vgrG genes was significantly changed under seven conditions. Among different stress conditions, high salt, and low temperature showed a higher effect on the expression of T6SS gene compared with host infection and other environmental conditions. As a first report, this study revealed an association of T6SS gene expression of the pathogen with the host infection, gene mutation, and some common environmental stresses. The results of this research can increase understanding of the biological function of T6SS in this economically-important pathogen of rice.

  11. Chelation of Membrane-Bound Cations by Extracellular DNA Activates the Type VI Secretion System in Pseudomonas aeruginosa.

    PubMed

    Wilton, Mike; Wong, Megan J Q; Tang, Le; Liang, Xiaoye; Moore, Richard; Parkins, Michael D; Lewenza, Shawn; Dong, Tao G

    2016-08-01

    Pseudomonas aeruginosa employs its type VI secretion system (T6SS) as a highly effective and tightly regulated weapon to deliver toxic molecules to target cells. T6SS-secreted proteins of P. aeruginosa can be detected in the sputum of cystic fibrosis (CF) patients, who typically present a chronic and polymicrobial lung infection. However, the mechanism of T6SS activation in the CF lung is not fully understood. Here we demonstrate that extracellular DNA (eDNA), abundant within the CF airways, stimulates the dynamics of the H1-T6SS cluster apparatus in Pseudomonas aeruginosa PAO1. Addition of Mg(2+) or DNase with eDNA abolished such activation, while treatment with EDTA mimicked the eDNA effect, suggesting that the eDNA-mediated effect is due to chelation of outer membrane-bound cations. DNA-activated H1-T6SS enables P. aeruginosa to nonselectively attack neighboring species regardless of whether or not it was provoked. Because of the importance of the T6SS in interspecies interactions and the prevalence of eDNA in the environments that P. aeruginosa inhabits, our report reveals an important adaptation strategy that likely contributes to the competitive fitness of P. aeruginosa in polymicrobial communities.

  12. Identifying Beneficial Qualities of Trichoderma parareesei for Plants

    PubMed Central

    Rubio, M. Belén; Quijada, Narciso M.; Pérez, Esclaudys; Domínguez, Sara; Hermosa, Rosa

    2014-01-01

    Trichoderma parareesei and Trichoderma reesei (teleomorph Hypocrea jecorina) produce cellulases and xylanases of industrial interest. Here, the anamorphic strain T6 (formerly T. reesei) has been identified as T. parareesei, showing biocontrol potential against fungal and oomycete phytopathogens and enhanced hyphal growth in the presence of tomato exudates or plant cell wall polymers in in vitro assays. A Trichoderma microarray was used to examine the transcriptomic changes in T6 at 20 h of interaction with tomato plants. Out of a total 34,138 Trichoderma probe sets deposited on the microarray, 250 showed a significant change of at least 2-fold in expression in the presence of tomato plants, with most of them being downregulated. T. parareesei T6 exerted beneficial effects on tomato plants in terms of seedling lateral root development, and in adult plants it improved defense against Botrytis cinerea and growth promotion under salt stress. Time course expression patterns (0 to 6 days) observed for defense-related genes suggest that T6 was able to prime defense responses in the tomato plants against biotic and abiotic stresses. Such responses undulated, with a maximum upregulation of the jasmonic acid (JA)/ethylene (ET)-related LOX1 and EIN2 genes and the salt tolerance SOS1 gene at 24 h and that of the salicylic acid (SA)-related PR-1 gene at 48 h after T6 inoculation. Our study demonstrates that the T. parareesei T6-tomato interaction is beneficial to both partners. PMID:24413597

  13. Structure and specificity of the Type VI secretion system ClpV-TssC interaction in enteroaggregative Escherichia coli

    PubMed Central

    Douzi, Badreddine; Brunet, Yannick R.; Spinelli, Silvia; Lensi, Valentine; Legrand, Pierre; Blangy, Stéphanie; Kumar, Anant; Journet, Laure; Cascales, Eric; Cambillau, Christian

    2016-01-01

    The Type VI secretion system (T6SS) is a versatile machine that delivers toxins into either eukaryotic or bacterial cells. It thus represents a key player in bacterial pathogenesis and inter-bacterial competition. Schematically, the T6SS can be viewed as a contractile tail structure anchored to the cell envelope. The contraction of the tail sheath propels the inner tube loaded with effectors towards the target cell. The components of the contracted tail sheath are then recycled by the ClpV AAA+ ATPase for a new cycle of tail elongation. The T6SS is widespread in Gram-negative bacteria and most of their genomes carry several copies of T6SS gene clusters, which might be activated in different conditions. Here, we show that the ClpV ATPases encoded within the two T6SS gene clusters of enteroaggregative Escherichia coli are not interchangeable and specifically participate to the activity of their cognate T6SS. Here we show that this specificity is dictated by interaction between the ClpV N-terminal domains and the N-terminal helices of their cognate TssC1 proteins. We also present the crystal structure of the ClpV1 N-terminal domain, alone or in complex with the TssC1 N-terminal peptide, highlighting the commonalities and diversities in the recruitment of ClpV to contracted sheaths. PMID:27698444

  14. Separate inputs modulate phosphorylation-dependent and -independent type VI secretion activation

    PubMed Central

    Silverman, Julie M.; Austin, Laura S.; Hsu, FoSheng; Hicks, Kevin G.; Hood, Rachel D.; Mougous, Joseph D.

    2013-01-01

    Summary Productive intercellular delivery of cargo by secretory systems requires exquisite temporal and spatial choreography. Our laboratory has demonstrated that the hemolysin co-regulated secretion island I (HSI-I)-encoded type VI secretion system (H1-T6SS) of Pseudomonas aeruginosa transfers effector proteins to other bacterial cells. The activity of these effectors requires cell contact-dependent delivery by the secretion apparatus, and thus their export is highly repressed under planktonic growth conditions. Here we define regulatory pathways that orchestrate efficient secretion by this system. We identified a T6S-associated protein, TagF, as a posttranslational repressor of the H1-T6SS. Strains activated by TagF derepression or stimulation of a previously identified threonine phosphorylation pathway (TPP) share the property of secretory ATPase recruitment to the T6S apparatus, yet display different effector output levels and genetic requirements for their export. We also found that the pathways respond to distinct stimuli; we identified surface growth as a physiological cue that activates the H1-T6SS exclusively through the TPP. Coordination of posttranslational triggering with cell contact-promoting growth conditions provides a mechanism for the T6SS to avoid wasteful release of effectors. PMID:22017253

  15. Visualization of the Serratia Type VI Secretion System Reveals Unprovoked Attacks and Dynamic Assembly

    PubMed Central

    Gerc, Amy J.; Diepold, Andreas; Trunk, Katharina; Porter, Michael; Rickman, Colin; Armitage, Judith P.; Stanley-Wall, Nicola R.; Coulthurst, Sarah J.

    2015-01-01

    Summary The Type VI secretion system (T6SS) is a bacterial nanomachine that fires toxic proteins into target cells. Deployment of the T6SS represents an efficient and widespread means by which bacteria attack competitors or interact with host organisms and may be triggered by contact from an attacking neighbor cell as a defensive strategy. Here, we use the opportunist pathogen Serratia marcescens and functional fluorescent fusions of key components of the T6SS to observe different subassemblies of the machinery simultaneously and on multiple timescales in vivo. We report that the localization and dynamic behavior of each of the components examined is distinct, revealing a multi-stage and dynamic assembly process for the T6SS machinery. We also show that the T6SS can assemble and fire without needing a cell contact trigger, defining an aggressive strategy that broadens target range and suggesting that activation of the T6SS is tailored to survival in specific niches. PMID:26387948

  16. Type VI secretion system: secretion by a contractile nanomachine

    PubMed Central

    Basler, Marek

    2015-01-01

    The type VI secretion systems (T6SS) are present in about a quarter of all Gram-negative bacteria. Several key components of T6SS are evolutionarily related to components of contractile nanomachines such as phages and R-type pyocins. The T6SS assembly is initiated by formation of a membrane complex that binds a phage-like baseplate with a sharp spike, and this is followed by polymerization of a long rigid inner tube and an outer contractile sheath. Effectors are preloaded onto the spike or into the tube during the assembly by various mechanisms. Contraction of the sheath releases an unprecedented amount of energy, which is used to thrust the spike and tube with the associated effectors out of the effector cell and across membranes of both bacterial and eukaryotic target cells. Subunits of the contracted sheath are recycled by T6SS-specific unfoldase to allow for a new round of assembly. Live-cell imaging has shown that the assembly is highly dynamic and its subcellular localization is in certain bacteria regulated with a remarkable precision. Through the action of effectors, T6SS has mainly been shown to contribute to pathogenicity and competition between bacteria. This review summarizes the knowledge that has contributed to our current understanding of T6SS mode of action. PMID:26370934

  17. Identifying beneficial qualities of Trichoderma parareesei for plants.

    PubMed

    Rubio, M Belén; Quijada, Narciso M; Pérez, Esclaudys; Domínguez, Sara; Monte, Enrique; Hermosa, Rosa

    2014-03-01

    Trichoderma parareesei and Trichoderma reesei (teleomorph Hypocrea jecorina) produce cellulases and xylanases of industrial interest. Here, the anamorphic strain T6 (formerly T. reesei) has been identified as T. parareesei, showing biocontrol potential against fungal and oomycete phytopathogens and enhanced hyphal growth in the presence of tomato exudates or plant cell wall polymers in in vitro assays. A Trichoderma microarray was used to examine the transcriptomic changes in T6 at 20 h of interaction with tomato plants. Out of a total 34,138 Trichoderma probe sets deposited on the microarray, 250 showed a significant change of at least 2-fold in expression in the presence of tomato plants, with most of them being downregulated. T. parareesei T6 exerted beneficial effects on tomato plants in terms of seedling lateral root development, and in adult plants it improved defense against Botrytis cinerea and growth promotion under salt stress. Time course expression patterns (0 to 6 days) observed for defense-related genes suggest that T6 was able to prime defense responses in the tomato plants against biotic and abiotic stresses. Such responses undulated, with a maximum upregulation of the jasmonic acid (JA)/ethylene (ET)-related LOX1 and EIN2 genes and the salt tolerance SOS1 gene at 24 h and that of the salicylic acid (SA)-related PR-1 gene at 48 h after T6 inoculation. Our study demonstrates that the T. parareesei T6-tomato interaction is beneficial to both partners.

  18. A multidrug resistance plasmid contains the molecular switch for type VI secretion in Acinetobacter baumannii

    PubMed Central

    Weber, Brent S.; Ly, Pek Man; Irwin, Joshua N.; Pukatzki, Stefan; Feldman, Mario F.

    2015-01-01

    Infections with Acinetobacter baumannii, one of the most troublesome and least studied multidrug-resistant superbugs, are increasing at alarming rates. A. baumannii encodes a type VI secretion system (T6SS), an antibacterial apparatus of Gram-negative bacteria used to kill competitors. Expression of the T6SS varies among different strains of A. baumannii, for which the regulatory mechanisms are unknown. Here, we show that several multidrug-resistant strains of A. baumannii harbor a large, self-transmissible resistance plasmid that carries the negative regulators for T6SS. T6SS activity is silenced in plasmid-containing, antibiotic-resistant cells, while part of the population undergoes frequent plasmid loss and activation of the T6SS. This activation results in T6SS-mediated killing of competing bacteria but renders A. baumannii susceptible to antibiotics. Our data show that a plasmid that has evolved to harbor antibiotic resistance genes plays a role in the differentiation of cells specialized in the elimination of competing bacteria. PMID:26170289

  19. A multidrug resistance plasmid contains the molecular switch for type VI secretion in Acinetobacter baumannii.

    PubMed

    Weber, Brent S; Ly, Pek Man; Irwin, Joshua N; Pukatzki, Stefan; Feldman, Mario F

    2015-07-28

    Infections with Acinetobacter baumannii, one of the most troublesome and least studied multidrug-resistant superbugs, are increasing at alarming rates. A. baumannii encodes a type VI secretion system (T6SS), an antibacterial apparatus of Gram-negative bacteria used to kill competitors. Expression of the T6SS varies among different strains of A. baumannii, for which the regulatory mechanisms are unknown. Here, we show that several multidrug-resistant strains of A. baumannii harbor a large, self-transmissible resistance plasmid that carries the negative regulators for T6SS. T6SS activity is silenced in plasmid-containing, antibiotic-resistant cells, while part of the population undergoes frequent plasmid loss and activation of the T6SS. This activation results in T6SS-mediated killing of competing bacteria but renders A. baumannii susceptible to antibiotics. Our data show that a plasmid that has evolved to harbor antibiotic resistance genes plays a role in the differentiation of cells specialized in the elimination of competing bacteria.

  20. The type VI protein secretion system contributes to biofilm formation and seed-to-seedling transmission of Acidovorax citrulli on melon.

    PubMed

    Tian, Yanli; Zhao, Yuqiang; Wu, Xinrong; Liu, Fengquan; Hu, Baishi; Walcott, Ronald R

    2015-01-01

    The type VI protein secretion system (T6SS) is essential for the virulence of several Gram-negative bacteria. In this study, we identified a T6SS gene cluster in Acidovorax citrulli, a plant-pathogenic bacterium that causes bacterial fruit blotch (BFB) of cucurbits. One T6SS cluster, of approximately 25 kb in length and comprising 17 genes, was found in the A. citrulli AAC00-1 genome. Seventeen A. citrulli mutants were generated, each with a deletion of a single T6SS core gene. There were significant differences in BFB seed-to-seedling transmission between wild-type A. citrulli strain, xjl12, and ΔvasD, ΔimpK, ΔimpJ and ΔimpF mutants (71.71%, 9.83%, 8.41%, 7.15% and 5.99% BFB disease index, respectively). In addition, we observed that these four mutants were reduced in melon seed colonization and biofilm formation; however, they were not affected in virulence when infiltrated into melon seedling tissues. There were no significant differences in BFB seed-to-seedling transmission, melon tissue colonization and biofilm formation between xjl12 and the other 13 T6SS mutants. Overall, our results indicate that T6SS plays a role in seed-to-seedling transmission of BFB on melon.

  1. Anticytoproliferative effect of Vitamin C on rat hepatic stellate cell

    PubMed Central

    Su, Min; Chao, Guo; Liang, Minqing; Song, Jianhua; Wu, Ka

    2016-01-01

    This study was conducted to investigate the potential therapeutical benefit of Vitamin (VC), a potent antioxidant, on suppressing proliferation of immortalized rat liver stellate cell line (HSC-T6) in vitro, and to discuss the underlying mechanism. HSC-T6 was co-treated with different concentrations of VC (50, 100, 200 μmol/L) on designed time points. Then, cell viability was assessed by using MTT analysis, and the changes of cytomorphology was observed with apoptosis-specific TUNEL and immunohistochemical stains, as well as the intracellular target genes was determined by using RT-PCR, respectively. As the outcomes, VC-treated HSC-T6 showed significantly inhibited cell growth in a dose-dependent manner when compared to the vehicle control. Cytologically, VC increased TUNEL-labeled positive cells in cultured HSC-T6, which the cell count was greater than vehicle control. Meanwhile, VC-treated HSC-T6 showed elevated immunoreactive for TGF-β1-labeled cells. Moreover, VC contributed to down-regulated expressions of intracellular c-myc, cyclin D1, mTOR mRNAs in HSC-T6. Collectively, these preliminary findings have demonstrated that VC-mediated anti-proliferative effect on HSCs is involved in molecular mechanisms of promoting apoptosis and blocking endogenous collagenation. PMID:27398165

  2. Gene Expression of Type VI Secretion System Associated with Environmental Survival in Acidovorax avenae subsp. avenae by Principle Component Analysis

    PubMed Central

    Cui, Zhouqi; Jin, Guoqiang; Li, Bin; Kakar, Kaleem Ullah; Ojaghian, Mohammad Reza; Wang, Yangli; Xie, Guanlin; Sun, Guochang

    2015-01-01

    Valine glycine repeat G (VgrG) proteins are regarded as one of two effectors of Type VI secretion system (T6SS) which is a complex multi-component secretion system. In this study, potential biological roles of T6SS structural and VgrG genes in a rice bacterial pathogen, Acidovorax avenae subsp. avenae (Aaa) RS-1, were evaluated under seven stress conditions using principle component analysis of gene expression. The results showed that growth of the pathogen was reduced by H2O2 and paraquat-induced oxidative stress, high salt, low temperature, and vgrG mutation, compared to the control. However, pathogen growth was unaffected by co-culture with a rice rhizobacterium Burkholderia seminalis R456. In addition, expression of 14 T6SS structural and eight vgrG genes was significantly changed under seven conditions. Among different stress conditions, high salt, and low temperature showed a higher effect on the expression of T6SS gene compared with host infection and other environmental conditions. As a first report, this study revealed an association of T6SS gene expression of the pathogen with the host infection, gene mutation, and some common environmental stresses. The results of this research can increase understanding of the biological function of T6SS in this economically-important pathogen of rice. PMID:26378528

  3. Chelation of Membrane-Bound Cations by Extracellular DNA Activates the Type VI Secretion System in Pseudomonas aeruginosa.

    PubMed

    Wilton, Mike; Wong, Megan J Q; Tang, Le; Liang, Xiaoye; Moore, Richard; Parkins, Michael D; Lewenza, Shawn; Dong, Tao G

    2016-08-01

    Pseudomonas aeruginosa employs its type VI secretion system (T6SS) as a highly effective and tightly regulated weapon to deliver toxic molecules to target cells. T6SS-secreted proteins of P. aeruginosa can be detected in the sputum of cystic fibrosis (CF) patients, who typically present a chronic and polymicrobial lung infection. However, the mechanism of T6SS activation in the CF lung is not fully understood. Here we demonstrate that extracellular DNA (eDNA), abundant within the CF airways, stimulates the dynamics of the H1-T6SS cluster apparatus in Pseudomonas aeruginosa PAO1. Addition of Mg(2+) or DNase with eDNA abolished such activation, while treatment with EDTA mimicked the eDNA effect, suggesting that the eDNA-mediated effect is due to chelation of outer membrane-bound cations. DNA-activated H1-T6SS enables P. aeruginosa to nonselectively attack neighboring species regardless of whether or not it was provoked. Because of the importance of the T6SS in interspecies interactions and the prevalence of eDNA in the environments that P. aeruginosa inhabits, our report reveals an important adaptation strategy that likely contributes to the competitive fitness of P. aeruginosa in polymicrobial communities. PMID:27271742

  4. Purification, crystallization and preliminary X-ray crystallographic analysis of TssL from Vibrio cholerae

    PubMed Central

    Jeong, Jae-Hee; Chang, Jeong Ho; Kim, Yeon-Gil

    2014-01-01

    The type VI secretion system (T6SS) is a macromolecular complex that is conserved in Gram-negative bacteria. The T6SS secretes effector proteins into recipient cells in a contact-dependent manner in order to accomplish cooperative and competitive interactions with the cells. Although the composition and mechanism of the T6SS have been intensively investigated across many Gram-negative bacteria, to date structural information on T6SS components from the important pathogen Vibrio cholerae has been rare. Here, the cloning, purification, crystallization and preliminary X-ray crystallographic analysis of the cytoplasmic domain of TssL, an inner membrane protein of the T6SS, from V. cholerae are reported. Diffraction data were collected to 1.5 Å resolution using synchrotron radiation. The crystal belonged to the hexagonal space group P61, with unit-cell parameters a = 78.4, b = 78.4, c = 49.5 Å. The successful structural characterization of TssL from V. cholerae will contribute to understanding the role of the membrane-associated subunits of the T6SS in more detail. PMID:25195905

  5. A Type VI secretion system encoding locus is required for Bordetella bronchiseptica immunomodulation and persistence in vivo.

    PubMed

    Weyrich, Laura S; Rolin, Olivier Y; Muse, Sarah J; Park, Jihye; Spidale, Nicholas; Kennett, Mary J; Hester, Sara E; Chen, Chun; Dudley, Edward G; Harvill, Eric T

    2012-01-01

    Type VI Secretion Systems (T6SSs) have been identified in numerous gram-negative pathogens, but the lack of a natural host infection model has limited analysis of T6SS contributions to infection and pathogenesis. Here, we describe disruption of a gene within locus encoding a putative T6SS in Bordetella bronchiseptica strain RB50, a respiratory pathogen that circulates in a broad range of mammals, including humans, domestic animals, and mice. The 26 gene locus encoding the B. bronchiseptica T6SS contains apparent orthologs to all known core genes and possesses thirteen novel genes. By generating an in frame deletion of clpV, which encodes a putative ATPase required for some T6SS-dependent protein secretion, we observe that ClpV contributes to in vitro macrophage cytotoxicity while inducing several eukaryotic proteins associated with apoptosis. Additionally, ClpV is required for induction of IL-1β, IL-6, IL-17, and IL-10 production in J774 macrophages infected with RB50. During infections in wild type mice, we determined that ClpV contributes to altered cytokine production, increased pathology, delayed lower respiratory tract clearance, and long term nasal cavity persistence. Together, these results reveal a natural host infection system in which to interrogate T6SS contributions to immunomodulation and pathogenesis.

  6. The involvement of the anticodon adjacent modified nucleoside N-(9-(BETA-D-ribofuranosyl) purine-6-ylcarbamoyl)-threonine in the biological function of E. coli tRNAile.

    PubMed Central

    Miller, J P; Hussain, Z; Schweizer, M P

    1976-01-01

    tRNAile was isolated from E. coli Cp 79 (leu-, arg-, thr-, his-, thiamin-, RCrel) which had been grown on a sub-optimal concentration of thr and was found to contain an average of 50% less N-[9-(beta-D-ribofuranosyl)- purin-6-ylcarbamoyl]threonine, t6Ado, than tRNAile from cells grown on an optimum concentration of thr and containing a normal complement of t6Ado. The two tRNA's were identical in their ability to be aminoacylated, to accept the 3'-terminal dinucleotide, and to form an ile-tRNAile-Tu-GTP complex. In contrast, the t6Ado-deficient-tRNA was significantly less efficient in binding to ribosomes compared to the normal tRNA. This difference was seen in the binding of deacylated tRNA and in the nonenzymatic and enzymatic binding of ile-tRNA, all in response to poly AUC. The t6Ado-deficient ile-tRNA demonstrated no binding at Mg2+ concentrations less than or equal to 10 mM, while the normal ile-tRNA bound at low Mg2+ concentrations. Tetracycline had the same effect on the normal as on the t6Ado-deficient ile-tRNA binding. As a control, the binding of phe-tRNA (which does not contain t6Ado) from normal and thr-starved cells in response to poly U was identical. It was concluded that t6Ado is required for proper codon-anticodon interaction. PMID:781621

  7. Burkholderia mallei and Burkholderia pseudomallei Cluster 1 Type VI Secretion System Gene Expression Is Negatively Regulated by Iron and Zinc

    PubMed Central

    Burtnick, Mary N.; Brett, Paul J.

    2013-01-01

    Burkholderia mallei is a facultative intracellular pathogen that causes glanders in humans and animals. Previous studies have demonstrated that the cluster 1 type VI secretion system (T6SS-1) expressed by this organism is essential for virulence in hamsters and is positively regulated by the VirAG two-component system. Recently, we have shown that T6SS-1 gene expression is up-regulated following internalization of this pathogen into phagocytic cells and that this system promotes multinucleated giant cell formation in infected tissue culture monolayers. In the present study, we further investigated the complex regulation of this important virulence factor. To assess T6SS-1 expression, B. mallei strains were cultured in various media conditions and Hcp1 production was analyzed by Western immunoblotting. Transcript levels of several VirAG-regulated genes (bimA, tssA, hcp1 and tssM) were also determined using quantitative real time PCR. Consistent with previous observations, T6SS-1 was not expressed during growth of B. mallei in rich media. Curiously, growth of the organism in minimal media (M9G) or minimal media plus casamino acids (M9CG) facilitated robust expression of T6SS-1 genes whereas growth in minimal media plus tryptone (M9TG) did not. Investigation of this phenomenon confirmed a regulatory role for VirAG in this process. Additionally, T6SS-1 gene expression was significantly down-regulated by the addition of iron and zinc to M9CG. Other genes under the control of VirAG did not appear to be as tightly regulated by these divalent metals. Similar results were observed for B. pseudomallei, but not for B. thailandensis. Collectively, our findings indicate that in addition to being positively regulated by VirAG, B. mallei and B. pseudomallei T6SS-1 gene expression is negatively regulated by iron and zinc. PMID:24146925

  8. Higher brain centers for social tasks in worker ants, Camponotus japonicus.

    PubMed

    Nishikawa, Michiko; Watanabe, Hidehiro; Yokohari, Fumio

    2012-05-01

    Ants, eusocial insects, have highly elaborate chemical communication systems using a wide variety of pheromones. In the carpenter ant, Camponotus japonicus, workers and queens have the female-specific basiconic sensilla on antennae. The antennal lobe, the primary processing center, in female carpenter ants contains about 480 glomeruli, which are divided into seven groups (T1–T7 glomeruli) based on sensory afferent tracts. The axons of sensory neurons in basiconic sensilla are thought to project to female-specific T6 glomeruli. Therefore, these sensilla and glomeruli are thought to relate to female-specific social tasks in the ants. By using dye filling into local neurons (LNs) and projection neurons (PNs) in the antennal lobe, we neuroanatomically revealed the existence of an isolated processing system for signals probably relating to social tasks in the worker ant. In the antennal lobe, two categories of glomeruli, T6 glomeruli and non-T6 glomeruli, are clearly segregated by LNs. Furthermore, axon terminals of uniglomerular PNs from the respective categories of glomeruli (T6 uni-PNs and non-T6 uni-PNs) are also segregated in the secondary olfactory centers, the calyces of the mushroom body and the lateral horn: T6 uni-PNs terminate in the outer layers of the basal ring and lip of mushroom body calyces and in the posterior region of the lateral horn, whereas non-T6 uni-PNs terminate in the middle and inner layers of the basal ring and lip and in the anterior region of the lateral horn. These findings suggest that information probably relating to social tasks might be isolated from other olfactory information and processed in a separate subsystem.

  9. Hybridization is limited between two lineages of freeze-resistant Trichinella during coinfection in a mouse model.

    PubMed

    Hecht, Luke B B; Thompson, Peter C; Lavin, Elizabeth S; Zarlenga, Dante S; Rosenthal, Benjamin M

    2016-03-01

    Hybridization between two closely related but distinct genetic lineages may lead to homogenization of the two lineages with potentially novel phenotypes, or selective pressure to avoid hybridization if the two lineages are truly distinct. Trichinella nativa and Trichinella T6 are zoonotic nematode parasites which can be distinguished genetically despite occasional hybridization. Here, using an experimental murine model, we attempt to determine whether there are barriers to hybridization when sizeable numbers of each lineage are allowed to coinfect a host. Two mice were independently infected with equal numbers of T. nativa and T6. The offspring of these coinfections were genotyped at two microsatellite loci and one mitochondrial locus capable of distinguishing T. nativa from T6 genotypes. Among larvae in the F1 generation, offspring of every possible mating were encountered. Most larvae (63.6%) derived from T. nativa×T. nativa matings, while 21.1% of offspring were the product of T6×T6 matings, and only 15.3% were hybrid offspring of T. nativa×T6 crosses, differing markedly from null expectations. In this experimental model, T. nativa and Trichinella T6 were able to mate, but ratios of offspring indicated pre- or post-zygotic barriers to hybridization that may include assortative mating, genetic incompatibilities, and/or differences in the fitness of offspring. These barriers would limit gene flow between these two lineages in a natural setting, serving as a barrier to their homogenization and promoting their persistence as distinct and separate entities. PMID:26721624

  10. Higher brain centers for social tasks in worker ants, Camponotus japonicus.

    PubMed

    Nishikawa, Michiko; Watanabe, Hidehiro; Yokohari, Fumio

    2012-05-01

    Ants, eusocial insects, have highly elaborate chemical communication systems using a wide variety of pheromones. In the carpenter ant, Camponotus japonicus, workers and queens have the female-specific basiconic sensilla on antennae. The antennal lobe, the primary processing center, in female carpenter ants contains about 480 glomeruli, which are divided into seven groups (T1–T7 glomeruli) based on sensory afferent tracts. The axons of sensory neurons in basiconic sensilla are thought to project to female-specific T6 glomeruli. Therefore, these sensilla and glomeruli are thought to relate to female-specific social tasks in the ants. By using dye filling into local neurons (LNs) and projection neurons (PNs) in the antennal lobe, we neuroanatomically revealed the existence of an isolated processing system for signals probably relating to social tasks in the worker ant. In the antennal lobe, two categories of glomeruli, T6 glomeruli and non-T6 glomeruli, are clearly segregated by LNs. Furthermore, axon terminals of uniglomerular PNs from the respective categories of glomeruli (T6 uni-PNs and non-T6 uni-PNs) are also segregated in the secondary olfactory centers, the calyces of the mushroom body and the lateral horn: T6 uni-PNs terminate in the outer layers of the basal ring and lip of mushroom body calyces and in the posterior region of the lateral horn, whereas non-T6 uni-PNs terminate in the middle and inner layers of the basal ring and lip and in the anterior region of the lateral horn. These findings suggest that information probably relating to social tasks might be isolated from other olfactory information and processed in a separate subsystem. PMID:22102363

  11. The icmF3 locus is involved in multiple adaptation- and virulence-related characteristics in Pseudomonas aeruginosa PAO1

    PubMed Central

    Lin, Jinshui; Cheng, Juanli; Chen, Keqi; Guo, Chenghao; Zhang, Weipeng; Yang, Xu; Ding, Wei; Ma, Li; Wang, Yao; Shen, Xihui

    2015-01-01

    The type VI secretion system (T6SS) is widely distributed in Gram-negative bacteria. Three separate T6SSs called H1-, H2-, and H3-T6SS have been discovered in Pseudomonas aeruginosa PAO1. Recent studies suggest that, in contrast to the H1-T6SS that targets prokaryotic cells, H2- and H3-T6SS are involved in interactions with both prokaryotic and eukaryotic cells. However, the detailed functions of T6SS components are still uncharacterized. The intracellular multiplication factor (IcmF) protein is conserved in type VI secretion systems (T6SS) of all different bacterial pathogens. Bioinformatic analysis revealed that IcmF3 in P. aeruginosa PAO1 is different from other IcmF homologs and may represent a new branch of these proteins with distinct functions. Herein, we have investigated the function of IcmF3 in this strain. We have shown that deletion of the icmF3 gene in P. aeruginosa PAO1 is associated with pleiotropic phenotypes. The icmF3 mutant has variant colony morphology and an hypergrowth phenotype in iron-limiting medium. Surprisingly, this mutant is also defective for the production of pyoverdine, as well as defects in swimming motility and virulence in a C. elegans worm model. The icmF3 mutant exhibits higher conjugation frequency than the wild type and increased biofilm formation on abiotic surfaces. Additionally, expression of two phenazine biosynthetic loci is increased in the icmF3 mutant, leading to the overproduction of pyocyanin. Finally, the mutant exhibits decreased susceptibility to aminoglycosides such as tobramycin and gentamicin. And the detected phenotypes can be restored completely or partially by trans complementation of wild type icmF3 gene. The pleiotropic effects observed upon icmF3 deletion demonstrate that icmF3 plays critical roles in both pathogenesis and environmental adaptation in P. aeruginosa PAO1. PMID:26484316

  12. The icmF3 locus is involved in multiple adaptation- and virulence-related characteristics in Pseudomonas aeruginosa PAO1.

    PubMed

    Lin, Jinshui; Cheng, Juanli; Chen, Keqi; Guo, Chenghao; Zhang, Weipeng; Yang, Xu; Ding, Wei; Ma, Li; Wang, Yao; Shen, Xihui

    2015-01-01

    The type VI secretion system (T6SS) is widely distributed in Gram-negative bacteria. Three separate T6SSs called H1-, H2-, and H3-T6SS have been discovered in Pseudomonas aeruginosa PAO1. Recent studies suggest that, in contrast to the H1-T6SS that targets prokaryotic cells, H2- and H3-T6SS are involved in interactions with both prokaryotic and eukaryotic cells. However, the detailed functions of T6SS components are still uncharacterized. The intracellular multiplication factor (IcmF) protein is conserved in type VI secretion systems (T6SS) of all different bacterial pathogens. Bioinformatic analysis revealed that IcmF3 in P. aeruginosa PAO1 is different from other IcmF homologs and may represent a new branch of these proteins with distinct functions. Herein, we have investigated the function of IcmF3 in this strain. We have shown that deletion of the icmF3 gene in P. aeruginosa PAO1 is associated with pleiotropic phenotypes. The icmF3 mutant has variant colony morphology and an hypergrowth phenotype in iron-limiting medium. Surprisingly, this mutant is also defective for the production of pyoverdine, as well as defects in swimming motility and virulence in a C. elegans worm model. The icmF3 mutant exhibits higher conjugation frequency than the wild type and increased biofilm formation on abiotic surfaces. Additionally, expression of two phenazine biosynthetic loci is increased in the icmF3 mutant, leading to the overproduction of pyocyanin. Finally, the mutant exhibits decreased susceptibility to aminoglycosides such as tobramycin and gentamicin. And the detected phenotypes can be restored completely or partially by trans complementation of wild type icmF3 gene. The pleiotropic effects observed upon icmF3 deletion demonstrate that icmF3 plays critical roles in both pathogenesis and environmental adaptation in P. aeruginosa PAO1. PMID:26484316

  13. Heat Treatment of AZ91D Mg-Al-Zn Alloy: Microstructural Evolution and Dynamic Response

    NASA Astrophysics Data System (ADS)

    Luong, Dung D.; Shunmugasamy, Vasanth Chakravarthy; Cox, James; Gupta, Nikhil; Rohatgi, Pradeep K.

    2013-11-01

    Magnesium alloys are attracting great interest from the automotive industry because of the potential for weight reduction. An AZ91D cast alloy was studied in the current work to understand the effect of heat treatment on the microstructure and dynamic compressive properties. The selected heat treatments include solution treatment (T4) and solution treatment followed by aging (T6). The as-cast alloy microstructure consists of intermetallic β-phase (Mg17Al12) precipitates surrounded by α + β lamellar eutectic in α-Mg solid solution. The AZ91D-T4 specimens showed small β-phase precipitates along the grain boundaries and regions of eutectic mixture. The T6 heat treatment causes the β-phase platelets in the α + β eutectic to grow and develop into β-precipitates. The difference in the phase morphology reflects into the mechanical properties. The Vickers hardness of the T6 heat-treated specimens was 3.6% higher than the as-cast alloy. The compressive yield strengths of T4 and T6 treated specimens were 1.3% and 43.1% higher than those of as-cast specimens. The high strain rate compression testing resulted in increase in the strength with strain rate for the T4 and T6 specimens. A maximum increase of 42% was observed in T6 specimen tested at a strain rate of 4,000/s in comparison to the quasi-static compression. Under high strain rate compression testing, the T6 heat-treated specimens showed failure of the β-precipitates resulting in increased energy absorption in comparison to the quasi-static compression.

  14. VgrG and PAAR Proteins Define Distinct Versions of a Functional Type VI Secretion System

    PubMed Central

    Cianfanelli, Francesca R.; Alcoforado Diniz, Juliana; Guo, Manman; De Cesare, Virginia; Trost, Matthias; Coulthurst, Sarah J.

    2016-01-01

    The Type VI secretion system (T6SS) is widespread among bacterial pathogens and acts as an effective weapon against competitor bacteria and eukaryotic hosts by delivering toxic effector proteins directly into target cells. The T6SS utilises a bacteriophage-like contractile machinery to expel a puncturing device based on a tube of Hcp topped with a VgrG spike, which can be extended by a final tip from a PAAR domain-containing protein. Effector proteins are believed to be delivered by specifically associating with particular Hcp, VgrG or PAAR proteins, either covalently (‘specialised’) or non-covalently (‘cargo’ effectors). Here we used the T6SS of the opportunistic pathogen Serratia marcescens, together with integratecd genetic, proteomic and biochemical approaches, to elucidate the role of specific VgrG and PAAR homologues in T6SS function and effector specificity, revealing new aspects and unexpected subtleties in effector delivery by the T6SS. We identified effectors, both cargo and specialised, absolutely dependent on a particular VgrG for delivery to target cells, and discovered that other cargo effectors can show a preference for a particular VgrG. The presence of at least one PAAR protein was found to be essential for T6SS function, consistent with designation as a ‘core’ T6SS component. We showed that specific VgrG-PAAR combinations are required to assemble a functional T6SS and that the three distinct VgrG-PAAR assemblies in S. marcescens exhibit distinct effector specificity and efficiency. Unexpectedly, we discovered that two different PAAR-containing Rhs proteins can functionally pair with the same VgrG protein. Showing that accessory EagR proteins are involved in these interactions, native VgrG-Rhs-EagR complexes were isolated and specific interactions between EagR and cognate Rhs proteins identified. This study defines an essential yet flexible role for PAAR proteins in the T6SS and highlights the existence of distinct versions of the

  15. A type VI secretion system is involved in Pseudomonas fluorescens bacterial competition.

    PubMed

    Decoin, Victorien; Barbey, Corinne; Bergeau, Dorian; Latour, Xavier; Feuilloley, Marc G J; Orange, Nicole; Merieau, Annabelle

    2014-01-01

    Protein secretion systems are crucial mediators of bacterial interactions with other organisms. Among them, the type VI secretion system (T6SS) is widespread in Gram-negative bacteria and appears to inject toxins into competitor bacteria and/or eukaryotic cells. Major human pathogens, such as Vibrio cholerae, Burkholderia and Pseudomonas aeruginosa, express T6SSs. Bacteria prevent self-intoxication by their own T6SS toxins by producing immunity proteins, which interact with the cognate toxins. We describe here an environmental P. fluorescens strain, MFE01, displaying an uncommon oversecretion of Hcp (hemolysin-coregulated protein) and VgrG (valine-glycine repeat protein G) into the culture medium. These proteins are characteristic components of a functional T6SS. The aim of this study was to attribute a role to this energy-consuming overexpression of the T6SS. The genome of MFE01 contains at least two hcp genes (hcp1 and hcp2), suggesting that there may be two putative T6SS clusters. Phenotypic studies have shown that MFE01 is avirulent against various eukaryotic cell models (amebas, plant or animal cell models), but has antibacterial activity against a wide range of competitor bacteria, including rhizobacteria and clinical bacteria. Depending on the prey cell, mutagenesis of the hcp2 gene in MFE01 abolishes or reduces this antibacterial killing activity. Moreover, the introduction of T6SS immunity proteins from S. marcescens, which is not killed by MFE01, protects E. coli against MFE01 killing. These findings suggest that the protein encoded by hcp2 is involved in the killing activity of MFE01 mediated by effectors of the T6SS targeting the peptidoglycan of Gram-negative bacteria. Our results indicate that MFE01 can protect potato tubers against Pectobacterium atrosepticum, which causes tuber soft rot. Pseudomonas fluorescens is often described as a major PGPR (plant growth-promoting rhizobacterium), and our results suggest that there may be a connection between

  16. A Pseudomonas aeruginosa type VI secretion phospholipase D effector targets both prokaryotic and eukaryotic cells.

    PubMed

    Jiang, Feng; Waterfield, Nicholas R; Yang, Jian; Yang, Guowei; Jin, Qi

    2014-05-14

    Widely found in animal and plant-associated proteobacteria, type VI secretion systems (T6SSs) are potentially capable of facilitating diverse interactions with eukaryotes and/or other bacteria. Pseudomonas aeruginosa encodes three distinct T6SS haemolysin coregulated protein (Hcp) secretion islands (H1, H2, and H3-T6SS), each involved in different aspects of the bacterium's interaction with other organisms. Here we describe the characterization of a P. aeruginosa H3-T6SS-dependent phospholipase D effector, PldB, and its three tightly linked cognate immunity proteins. PldB targets the periplasm of prokaryotic cells and exerts an antibacterial activity. Surprisingly, PldB also facilitates intracellular invasion of host eukaryotic cells by activation of the PI3K/Akt pathway, revealing it to be a trans-kingdom effector. Our findings imply a potentially widespread T6SS-mediated mechanism, which deploys a single phospholipase effector to influence both prokaryotic cells and eukaryotic hosts.

  17. Sequence dependent electrophoretic mobilities and melting temperatures for A-T containing oligodeoxyribonucleotides.

    PubMed Central

    Wilson, W D; Zuo, E T; Jones, R L; Zon, G L; Baumstark, B R

    1987-01-01

    The electrophoretic mobilities and thermal melting properties of self complementary A-T containing dodecamer oligodeoxyribonucleotides have been investigated as a function of solution conditions. The oligomers contained tracts of nonalternating A-T base pairs of 2 (d(A2T2)3), 3 (d(A3T3)2), and 6 (d(A6T6] as well as the fully alternating (d(A-T)6) sequence. The melting temperature increased with the length of the nonalternating sequence and was approximately 12 degrees C higher in the d(A6T6) sequence than in the alternating oligomer. Under denaturing conditions all oligomers had the same electrophoretic mobility on acrylamide gels. Under conditions which favor duplex formation, the oligomers exhibited significant sequence dependent mobility differences. The mobilities of two oligomers, d(A-T)6 and d(A6-T6), were approximately equal and were less than those of the other oligonucleotides. The greatest mobility was observed for d(A2T2)3. These results are best explained by a model which requires bending at a junction of two or more continuous A or T bases with another sequence. Images PMID:3822802

  18. TssB is essential for virulence and required for type VI secretion system in Ralstonia solanacearum.

    PubMed

    Zhang, Liqing; Xu, Jingsheng; Xu, Jin; Zhang, Hao; He, Liyuan; Feng, Jie

    2014-09-01

    The type VI secretion system (T6SS) is recently discovered machinery in Gram-negative bacteria for translocation of proteins and also is required for full virulence. TssB is a highly conserved protein among the T6SSs, and indispensable for composition and function of T6S. The plant pathogenic bacterium Ralstonia solanacearum also harbours T6SS gene clusters, and a homologue of TssB, hereafter designated as TssBRS, but up to date its characterization and function remain unclear. In this study, we showed that TssBRS of R. solanacearum was required for secretion of Hcp, the haemolysin coregulated protein and a hallmark of T6S pathway. Deletion of tssBRS in R. solanacearum GMI1000 strain resulted in defect of biofilm formation, and the expression of the flagella operon is decreased, leading to decreased motility. More importantly, tssBRS mutant strain had significantly attenuated its virulence on tomato plants. TssB is essential for virulence and required for type VI secretion system in R. solanacearum.

  19. Quorum sensing control of Type VI secretion factors restricts the proliferation of quorum-sensing mutants

    PubMed Central

    Majerczyk, Charlotte; Schneider, Emily; Greenberg, E Peter

    2016-01-01

    Burkholderia thailandensis uses acyl-homoserine lactone-mediated quorum sensing systems to regulate hundreds of genes. Here we show that cell-cell contact-dependent type VI secretion (T6S) toxin-immunity systems are among those activated by quorum sensing in B. thailandensis. We also demonstrate that T6S is required to constrain proliferation of quorum sensing mutants in colony cocultures of a BtaR1 quorum-sensing signal receptor mutant and its parent. However, the BtaR1 mutant is not constrained by and outcompetes its parent in broth coculture, presumably because no cell contact occurs and there is a metabolic cost associated with quorum sensing gene activation. The increased fitness of the wild type over the BtaR1 mutant during agar surface growth is dependent on an intact T6SS-1 apparatus. Thus, quorum sensing activates B. thailandensis T6SS-1 growth inhibition and this control serves to police and constrain quorum-sensing mutants. This work defines a novel role for T6SSs in intraspecies mutant control. DOI: http://dx.doi.org/10.7554/eLife.14712.001 PMID:27183270

  20. TssA forms a gp6-like ring attached to the type VI secretion sheath.

    PubMed

    Planamente, Sara; Salih, Osman; Manoli, Eleni; Albesa-Jové, David; Freemont, Paul S; Filloux, Alain

    2016-08-01

    The type VI secretion system (T6SS) is a supra-molecular bacterial complex that resembles phage tails. It is a killing machine which fires toxins into target cells upon contraction of its TssBC sheath. Here, we show that TssA1 is a T6SS component forming dodecameric ring structures whose dimensions match those of the TssBC sheath and which can accommodate the inner Hcp tube. The TssA1 ring complex binds the T6SS sheath and impacts its behaviour in vivo In the phage, the first disc of the gp18 sheath sits on a baseplate wherein gp6 is a dodecameric ring. We found remarkable sequence and structural similarities between TssA1 and gp6 C-termini, and propose that TssA1 could be a baseplate component of the T6SS Furthermore, we identified similarities between TssK1 and gp8, the former interacting with TssA1 while the latter is found in the outer radius of the gp6 ring. These observations, combined with similarities between TssF and gp6N-terminus or TssG and gp53, lead us to propose a comparative model between the phage baseplate and the T6SS. PMID:27288401

  1. Fatigue Properties of Cast Magnesium Wheels

    NASA Astrophysics Data System (ADS)

    Li, Zhenming; Luo, Alan A.; Wang, Qigui; Peng, Liming; Zhang, Peng

    2016-08-01

    This paper investigates the fatigue properties and deformation behavior of a newly developed Mg-2.96Nd-0.21Zn-0.39Zr magnesium alloy wheel in both as-cast and T6 conditions. Compared with the as-cast alloy, the T6-treated alloy shows a significant increase in fatigue strength and cyclic stress amplitude. This is believed to be attributed to the change of defect type from porosity to oxides and the increased matrix strength in the T6 (peak-aged) condition. For the as-cast alloy wheel, fatigue failure mainly originated from the cast defects including porosity, oxide film, and inclusion at or near the sample surface. In the T6-treated alloy, however, oxides and inclusions or slip bands initiate the fatigue cracks. Solution treatment appears to reduce or eliminate the shrinkage porosity because of grain growth and dissolution of as-cast eutectic phases in the grain boundaries. The cyclic stress amplitude of the as-cast alloy increases with increasing the number of cycles, while the T6-treated alloy shows cyclic softening after the stress reaches a maximum value. The Coffin-Manson law and Basquin equation can be used to evaluate the life of low cycle fatigue. The developed long crack model and multi-scale fatigue (MSF) models can be used to predict high-cycle fatigue life of the Mg-2.96Nd-0.21Zn-0.39Zr alloys with or without casting defects.

  2. Type VI Secretion System Substrates Are Transferred and Reused among Sister Cells.

    PubMed

    Vettiger, Andrea; Basler, Marek

    2016-09-22

    Bacterial type VI secretion system (T6SS) is a nanomachine that works similarly to a speargun. Rapid contraction of a sling (sheath) drives a long shaft (Hcp) with a sharp tip and associated effectors through the target cell membrane. We show that the amount and composition of the tip regulates initiation of full-length sheath assembly and low amount of available Hcp decreases sheath length. Importantly, we show that both tip and Hcp are exchanged by T6SS among by-standing cells within minutes of initial cell-cell contact. The translocated proteins are reused for new T6SS assemblies suggesting that tip and Hcp reach the cytosol of target cells. The efficiency of protein translocation depends on precise aiming of T6SS at the target cells. This interbacterial protein complementation can support T6SS activity in sister cells with blocked protein synthesis and also allows cooperation between strains to increase their potential to kill competition. VIDEO ABSTRACT.

  3. Quorum sensing control of Type VI secretion factors restricts the proliferation of quorum-sensing mutants.

    PubMed

    Majerczyk, Charlotte; Schneider, Emily; Greenberg, E Peter

    2016-05-16

    Burkholderia thailandensis uses acyl-homoserine lactone-mediated quorum sensing systems to regulate hundreds of genes. Here we show that cell-cell contact-dependent type VI secretion (T6S) toxin-immunity systems are among those activated by quorum sensing in B. thailandensis. We also demonstrate that T6S is required to constrain proliferation of quorum sensing mutants in colony cocultures of a BtaR1 quorum-sensing signal receptor mutant and its parent. However, the BtaR1 mutant is not constrained by and outcompetes its parent in broth coculture, presumably because no cell contact occurs and there is a metabolic cost associated with quorum sensing gene activation. The increased fitness of the wild type over the BtaR1 mutant during agar surface growth is dependent on an intact T6SS-1 apparatus. Thus, quorum sensing activates B. thailandensis T6SS-1 growth inhibition and this control serves to police and constrain quorum-sensing mutants. This work defines a novel role for T6SSs in intraspecies mutant control.

  4. Prevalence of Type VI Secretion System in Spanish Campylobacter jejuni Isolates.

    PubMed

    Ugarte-Ruiz, M; Stabler, R A; Domínguez, L; Porrero, M C; Wren, B W; Dorrell, N; Gundogdu, O

    2015-11-01

    Infections from Campylobacter jejuni pose a serious public health problem and are now considered the leading cause of foodborne bacterial gastroenteritis throughout the world. Sequencing of C. jejuni genomes has previously allowed a number of loci to be identified, which encode virulence factors that aid survival and pathogenicity. Recently, a Type VI secretion system (T6SS) consisting of 13 conserved genes was described in C. jejuni strains and recognised to promote pathogenicity and adaptation to the environment. In this study, we determined the presence of this T6SS in 63 Spanish C. jejuni isolates from the food chain and urban effluents using whole-genome sequencing. Our findings demonstrated that nine (14%) strains harboured the 13 ORFs found in prototype strain C. jejuni 108. Further studies will be necessary to determine the prevalence and importance of T6SS-positive C. jejuni strains.

  5. Type VI secretion effectors: poisons with a purpose

    PubMed Central

    Russell, Alistair B.; Peterson, S. Brook; Mougous, Joseph D.

    2014-01-01

    The type VI secretion system (T6SS) mediates interactions between a diverse range of Gram-negative bacterial species. Recent studies have led to a drastic increase in the number of characterized T6SS effector proteins and produced a more complete and nuanced view of the adaptive significance of the system. While the system is most often implicated in antagonism, in this review we consider the case for its involvement in both antagonistic and non-antagonistic behaviors. Clarifying the roles that T6S plays in microbial communities will contribute to broader efforts to understand the importance of microbial interactions in maintaining human and environmental health, and will inform efforts to manipulate these interactions for therapeutic or environmental benefit. PMID:24384601

  6. Type VI secretion apparatus and phage tail-associated protein complexes share a common evolutionary origin

    SciTech Connect

    Leiman, Petr G.; Basler, Marek; Ramagopal, Udupi A.; Bonanno, Jeffrey B.; Sauder, J. Michael; Pukatzki, Stefan; Burley, Stephen K.; Almo, Steven C.; Mekalanos, John J.

    2009-04-22

    Protein secretion is a common property of pathogenic microbes. Gram-negative bacterial pathogens use at least 6 distinct extracellular protein secretion systems to export proteins through their multilayered cell envelope and in some cases into host cells. Among the most widespread is the newly recognized Type VI secretion system (T6SS) which is composed of 15--20 proteins whose biochemical functions are not well understood. Using crystallographic, biochemical, and bioinformatic analyses, we identified 3 T6SS components, which are homologous to bacteriophage tail proteins. These include the tail tube protein; the membrane-penetrating needle, situated at the distal end of the tube; and another protein associated with the needle and tube. We propose that T6SS is a multicomponent structure whose extracellular part resembles both structurally and functionally a bacteriophage tail, an efficient machine that translocates proteins and DNA across lipid membranes into cells.

  7. An ABC transporter and an outer membrane lipoprotein participate in posttranslational activation of type VI secretion in Pseudomonas aeruginosa.

    PubMed

    Casabona, Maria G; Silverman, Julie M; Sall, Khady M; Boyer, Frédéric; Couté, Yohann; Poirel, Jessica; Grunwald, Didier; Mougous, Joseph D; Elsen, Sylvie; Attree, Ina

    2013-02-01

    Pseudomonas aeruginosa is capable of injecting protein toxins into other bacterial cells through one of its three type VI secretion systems (T6SSs). The activity of this T6SS is tightly regulated on the posttranslational level by phosphorylation-dependent and -independent pathways. The phosphorylation-dependent pathway consists of a Threonine kinase/phosphatase pair (PpkA/PppA) that acts on a forkhead domain-containing protein, Fha1, and a periplasmic protein, TagR, that positively regulates PpkA. In the present work, we biochemically and functionally characterize three additional proteins of the phosphorylation-dependent regulatory cascade that controls T6S activation: TagT, TagS and TagQ. We show that similar to TagR, these proteins act upstream of the PpkA/PppA checkpoint and influence phosphorylation of Fha1 and, apparatus assembly and effector export. Localization studies demonstrate that TagQ is an outer membrane lipoprotein and TagR is associated with the outer membrane. Consistent with their homology to lipoprotein outer membrane localization (Lol) components, TagT and TagS form a stable inner membrane complex with ATPase activity. However, we find that outer membrane association of T6SS lipoproteins TagQ and TssJ1, and TagR, is unaltered in a ΔtagTS background. Notably, we found that TagQ is indispensible for anchoring of TagR to the outer membrane fraction. As T6S-dependent fitness of P. aeruginosa requires TagT, S, R and Q, we conclude that these proteins likely participate in a trans-membrane signalling pathway that promotes H1-T6SS activity under optimal environmental conditions. PMID:22765374

  8. Contribution of the type VI secretion system encoded in SPI-19 to chicken colonization by Salmonella enterica serotypes Gallinarum and Enteritidis.

    PubMed

    Blondel, Carlos J; Yang, Hee-Jeong; Castro, Benjamín; Chiang, Sebastián; Toro, Cecilia S; Zaldívar, Mercedes; Contreras, Inés; Andrews-Polymenis, Helene L; Santiviago, Carlos A

    2010-07-22

    Salmonella Gallinarum is a pathogen with a host range specific to poultry, while Salmonella Enteritidis is a broad host range pathogen that colonizes poultry sub-clinically but is a leading cause of gastrointestinal salmonellosis in humans and many other species. Despite recent advances in our understanding of the complex interplay between Salmonella and their hosts, the molecular basis of host range restriction and unique pathobiology of Gallinarum remain largely unknown. Type VI Secretion System (T6SS) represents a new paradigm of protein secretion that is critical for the pathogenesis of many gram-negative bacteria. We recently identified a putative T6SS in the Salmonella Pathogenicity Island 19 (SPI-19) of Gallinarum. In Enteritidis, SPI-19 is a degenerate element that has lost most of the T6SS functions encoded in the island. In this work, we studied the contribution of SPI-19 to the colonization of Salmonella Gallinarum strain 287/91 in chickens. Non-polar deletion mutants of SPI-19 and the clpV gene, an essential T6SS component, colonized the ileum, ceca, liver and spleen of White Leghorn chicks poorly compared to the wild-type strain after oral inoculation. Return of SPI-19 to the DeltaSPI-19 mutant, using VEX-Capture, complemented this colonization defect. In contrast, transfer of SPI-19 from Gallinarum to Enteritidis resulted in transient increase in the colonization of the ileum, liver and spleen at day 1 post-infection, but at days 3 and 5 post-infection a strong colonization defect of the gut and internal organs of the experimentally infected chickens was observed. Our data indicate that SPI-19 and the T6SS encoded in this region contribute to the colonization of the gastrointestinal tract and internal organs of chickens by Salmonella Gallinarum and suggest that degradation of SPI-19 T6SS in Salmonella Enteritidis conferred an advantage in colonization of the avian host.

  9. Contribution of the Type VI Secretion System Encoded in SPI-19 to Chicken Colonization by Salmonella enterica Serotypes Gallinarum and Enteritidis

    PubMed Central

    Blondel, Carlos J.; Yang, Hee-Jeong; Castro, Benjamín; Chiang, Sebastián; Toro, Cecilia S.; Zaldívar, Mercedes; Contreras, Inés; Andrews-Polymenis, Helene L.; Santiviago, Carlos A.

    2010-01-01

    Salmonella Gallinarum is a pathogen with a host range specific to poultry, while Salmonella Enteritidis is a broad host range pathogen that colonizes poultry sub-clinically but is a leading cause of gastrointestinal salmonellosis in humans and many other species. Despite recent advances in our understanding of the complex interplay between Salmonella and their hosts, the molecular basis of host range restriction and unique pathobiology of Gallinarum remain largely unknown. Type VI Secretion System (T6SS) represents a new paradigm of protein secretion that is critical for the pathogenesis of many Gram-negative bacteria. We recently identified a putative T6SS in the Salmonella Pathogenicity Island 19 (SPI-19) of Gallinarum. In Enteritidis, SPI-19 is a degenerate element that has lost most of the T6SS functions encoded in the island. In this work, we studied the contribution of SPI-19 to the colonization of Salmonella Gallinarum strain 287/91 in chickens. Non-polar deletion mutants of SPI-19 and the clpV gene, an essential T6SS component, colonized the ileum, ceca, liver and spleen of White Leghorn chicks poorly compared to the wild-type strain after oral inoculation. Return of SPI-19 to the ΔSPI-19 mutant, using VEX-Capture, complemented this colonization defect. In contrast, transfer of SPI-19 from Gallinarum to Enteritidis resulted in transient increase in the colonization of the ileum, liver and spleen at day 1 post-infection, but at days 3 and 5 post-infection a strong colonization defect of the gut and internal organs of the experimentally infected chickens was observed. Our data indicate that SPI-19 and the T6SS encoded in this region contribute to the colonization of the gastrointestinal tract and internal organs of chickens by Salmonella Gallinarum and suggest that degradation of SPI-19 T6SS in Salmonella Enteritidis conferred an advantage in colonization of the avian host. PMID:20661437

  10. The Type VI Secretion TssEFGK-VgrG Phage-Like Baseplate Is Recruited to the TssJLM Membrane Complex via Multiple Contacts and Serves As Assembly Platform for Tail Tube/Sheath Polymerization

    PubMed Central

    Boyer, Frédéric; Douzi, Badreddine; Cascales, Eric

    2015-01-01

    The Type VI secretion system (T6SS) is a widespread weapon dedicated to the delivery of toxin proteins into eukaryotic and prokaryotic cells. The 13 T6SS subunits assemble a cytoplasmic contractile structure anchored to the cell envelope by a membrane-spanning complex. This structure is evolutionarily, structurally and functionally related to the tail of contractile bacteriophages. In bacteriophages, the tail assembles onto a protein complex, referred to as the baseplate, that not only serves as a platform during assembly of the tube and sheath, but also triggers the contraction of the sheath. Although progress has been made in understanding T6SS assembly and function, the composition of the T6SS baseplate remains mostly unknown. Here, we report that six T6SS proteins–TssA, TssE, TssF, TssG, TssK and VgrG–are required for proper assembly of the T6SS tail tube, and a complex between VgrG, TssE,-F and-G could be isolated. In addition, we demonstrate that TssF and TssG share limited sequence homologies with known phage components, and we report the interaction network between these subunits and other baseplate and tail components. In agreement with the baseplate being the assembly platform for the tail, fluorescence microscopy analyses of functional GFP-TssF and TssK-GFP fusion proteins show that these proteins assemble stable and static clusters on which the sheath polymerizes. Finally, we show that recruitment of the baseplate to the apparatus requires initial positioning of the membrane complex and contacts between TssG and the inner membrane TssM protein. PMID:26460929

  11. Structure of the Trehalose-6-phosphate Phosphatase from Brugia malayi Reveals Key Design Principles for Anthelmintic Drugs

    PubMed Central

    Farelli, Jeremiah D.; Galvin, Brendan D.; Li, Zhiru; Liu, Chunliang; Aono, Miyuki; Garland, Megan; Hallett, Olivia E.; Causey, Thomas B.; Ali-Reynolds, Alana; Saltzberg, Daniel J.; Carlow, Clotilde K. S.; Dunaway-Mariano, Debra; Allen, Karen N.

    2014-01-01

    Parasitic nematodes are responsible for devastating illnesses that plague many of the world's poorest populations indigenous to the tropical areas of developing nations. Among these diseases is lymphatic filariasis, a major cause of permanent and long-term disability. Proteins essential to nematodes that do not have mammalian counterparts represent targets for therapeutic inhibitor discovery. One promising target is trehalose-6-phosphate phosphatase (T6PP) from Brugia malayi. In the model nematode Caenorhabditis elegans, T6PP is essential for survival due to the toxic effect(s) of the accumulation of trehalose 6-phosphate. T6PP has also been shown to be essential in Mycobacterium tuberculosis. We determined the X-ray crystal structure of T6PP from B. malayi. The protein structure revealed a stabilizing N-terminal MIT-like domain and a catalytic C-terminal C2B-type HAD phosphatase fold. Structure-guided mutagenesis, combined with kinetic analyses using a designed competitive inhibitor, trehalose 6-sulfate, identified five residues important for binding and catalysis. This structure-function analysis along with computational mapping provided the basis for the proposed model of the T6PP-trehalose 6-phosphate complex. The model indicates a substrate-binding mode wherein shape complementarity and van der Waals interactions drive recognition. The mode of binding is in sharp contrast to the homolog sucrose-6-phosphate phosphatase where extensive hydrogen-bond interactions are made to the substrate. Together these results suggest that high-affinity inhibitors will be bi-dentate, taking advantage of substrate-like binding to the phosphoryl-binding pocket while simultaneously utilizing non-native binding to the trehalose pocket. The conservation of the key residues that enforce the shape of the substrate pocket in T6PP enzymes suggest that development of broad-range anthelmintic and antibacterial therapeutics employing this platform may be possible. PMID:24992307

  12. Identification of an Inhibitory Alcohol Binding Site in GABAA ρ1 Receptors.

    PubMed

    Borghese, Cecilia M; Ruiz, Carlos I; Lee, Ui S; Cullins, Madeline A; Bertaccini, Edward J; Trudell, James R; Harris, R Adron

    2016-01-20

    Alcohols inhibit γ-aminobutyric acid type A ρ1 receptor function. After introducing mutations in several positions of the second transmembrane helix in ρ1, we studied the effects of ethanol and hexanol on GABA responses using two-electrode voltage clamp electrophysiology in Xenopus laevis oocytes. The 6' mutations produced the following effects on ethanol and hexanol responses: small increase or no change (T6'M), increased inhibition (T6'V), and small potentiation (T6'Y and T6'F). The 5' mutations produced mainly increases in hexanol inhibition. Other mutations produced small (3' and 9') or no changes (2' and L277 in the first transmembrane domain) in alcohol effects. These results suggest an inhibitory alcohol binding site near the 6' position. Homology models of ρ1 receptors based on the X-ray structure of GluCl showed that the 2', 5', 6', and 9' residues were easily accessible from the ion pore, with 5' and 6' residues from neighboring subunits facing each other; L3' and L277 also faced the neighboring subunit. We tested ethanol through octanol on single and double mutated ρ1 receptors [ρ1(I15'S), ρ1(T6'Y), and ρ1(T6'Y,I15'S)] to further characterize the inhibitory alcohol pocket in the wild-type ρ1 receptor. The pocket can only bind relatively short-chain alcohols and is eliminated by introducing Y in the 6' position. Replacing the bulky 15' residue with a smaller side chain introduced a potentiating binding site, more sensitive to long-chain than to short-chain alcohols. In conclusion, the net alcohol effect on the ρ1 receptor is determined by the sum of its actions on inhibitory and potentiating sites.

  13. Understanding the sequential activation of Type III and Type VI Secretion Systems in Salmonella typhimurium using Boolean modeling

    PubMed Central

    2013-01-01

    Background Three pathogenicity islands, viz. SPI-1 (Salmonella pathogenicity island 1), SPI-2 (Salmonella pathogenicity island 2) and T6SS (Type VI Secretion System), present in the genome of Salmonella typhimurium have been implicated in the virulence of the pathogen. While the regulation of SPI-1 and SPI-2 (both encoding components of the Type III Secretion System - T3SS) are well understood, T6SS regulation is comparatively less studied. Interestingly, inter-connections among the regulatory elements of these three virulence determinants have also been suggested to be essential for successful infection. However, till date, an integrated view of gene regulation involving the regulators of these three secretion systems and their cross-talk is not available. Results In the current study, relevant regulatory information available from literature have been integrated into a single Boolean network, which portrays the dynamics of T3SS (SPI-1 and SPI-2) and T6SS mediated virulence. Some additional regulatory interactions involving a two-component system response regulator YfhA have also been predicted and included in the Boolean network. These predictions are aimed at deciphering the effects of osmolarity on T6SS regulation, an aspect that has been suggested in earlier studies, but the mechanism of which was hitherto unknown. Simulation of the regulatory network was able to recreate in silico the experimentally observed sequential activation of SPI-1, SPI-2 and T6SS. Conclusions The present study integrates relevant gene regulatory data (from literature and our prediction) into a single network, representing the cross-communication between T3SS (SPI-1 and SPI-2) and T6SS. This holistic view of regulatory interactions is expected to improve the current understanding of pathogenesis of S. typhimurium. PMID:24079299

  14. Validation of a six second cycle test for the determination of peak power output.

    PubMed

    Herbert, Peter; Sculthorpe, Nick; Baker, Julien S; Grace, Fergal M

    2015-01-01

    The present study examined the agreement between peak power output during a standard Wingate anaerobic test (WAnT) and a six second 'all-out' test on a Wattbike Pro. Nine males (40.7 ± 19.4 yrs, 1.76 ± 0.03 cm, 82.11 ± 8.9 kg) underwent three testing protocols on separate days. The protocols consisted 30 second WAnT (WAnT30), a modified WAnT over 6 seconds (WAnT6) and a 6 second peak power test (PPT6). PPT6 was correlated with WAnT30 (r = 0.9; p < 0.001) with a mean bias of 105 W. PPT6 correlated with WAnT6 (r = 0.95; p < 0.001) with a mean bias of 74 W. WAnT6 correlated with WAnT30 (r = 0.99; p < 0.001) with a mean bias of 31 W. There was no difference in time to peak power between any trial. PPT6 resulted in significantly greater power outputs than in WAnT30 and WAnT6 (p < 0.001). We conclude that PPT6 and WAnT6 are valid measures of peak power output compared with WAnT30. This identifies that PPT6 and WAnT6 as short duration 'all-out' tests that have practical applications for researchers and coaches who wish to assess peak power output without the fatiguing effects associated with a standard WAnT.

  15. Structure of the trehalose-6-phosphate phosphatase from Brugia malayi reveals key design principles for anthelmintic drugs.

    PubMed

    Farelli, Jeremiah D; Galvin, Brendan D; Li, Zhiru; Liu, Chunliang; Aono, Miyuki; Garland, Megan; Hallett, Olivia E; Causey, Thomas B; Ali-Reynolds, Alana; Saltzberg, Daniel J; Carlow, Clotilde K S; Dunaway-Mariano, Debra; Allen, Karen N

    2014-07-01

    Parasitic nematodes are responsible for devastating illnesses that plague many of the world's poorest populations indigenous to the tropical areas of developing nations. Among these diseases is lymphatic filariasis, a major cause of permanent and long-term disability. Proteins essential to nematodes that do not have mammalian counterparts represent targets for therapeutic inhibitor discovery. One promising target is trehalose-6-phosphate phosphatase (T6PP) from Brugia malayi. In the model nematode Caenorhabditis elegans, T6PP is essential for survival due to the toxic effect(s) of the accumulation of trehalose 6-phosphate. T6PP has also been shown to be essential in Mycobacterium tuberculosis. We determined the X-ray crystal structure of T6PP from B. malayi. The protein structure revealed a stabilizing N-terminal MIT-like domain and a catalytic C-terminal C2B-type HAD phosphatase fold. Structure-guided mutagenesis, combined with kinetic analyses using a designed competitive inhibitor, trehalose 6-sulfate, identified five residues important for binding and catalysis. This structure-function analysis along with computational mapping provided the basis for the proposed model of the T6PP-trehalose 6-phosphate complex. The model indicates a substrate-binding mode wherein shape complementarity and van der Waals interactions drive recognition. The mode of binding is in sharp contrast to the homolog sucrose-6-phosphate phosphatase where extensive hydrogen-bond interactions are made to the substrate. Together these results suggest that high-affinity inhibitors will be bi-dentate, taking advantage of substrate-like binding to the phosphoryl-binding pocket while simultaneously utilizing non-native binding to the trehalose pocket. The conservation of the key residues that enforce the shape of the substrate pocket in T6PP enzymes suggest that development of broad-range anthelmintic and antibacterial therapeutics employing this platform may be possible.

  16. Methanogenic activity inhibition by increasing the linear alkylbenzene sulfonate (LAS) concentration.

    PubMed

    Souza, Luiza F C; Florencio, Lourdinha; Gavazza, Savia; Kato, Mario T

    2016-07-01

    The effect of the initial concentration of linear alkylbenzene sulfonate (LAS) on specific methanogenic activity (SMA) was investigated in this work. Six anaerobic flasks reactors with 1 L of total volume were inoculated with anaerobic sludge (2 g VSS L(-1)). The reactors were assayed for 42 days, and fed with volatile fatty acids, nutrients, and LAS. The initial LAS concentrations were 0, 10, 30, 50, 75, and 100 mg L(-1) for the treatment flasks T1 (control), T2, T3, T4, T5, and T6, respectively. When compared with T1, T2 exhibited a 30% reduction in maximum SMA and total methane production (TMP). In treatment T3 through T6, the reductions were 44-97% (T3-T6) for SMA, and 30-90% (T3-T6) for TMP. Total LAS removal increased following the increase in the initial LAS concentration (from 36% at T1 to 76% at T6), primarily due to the high degree of sludge adsorption. LAS biodegradation also occurred (32% in all treatments), although this was most likely associated with the formation of non-methane intermediates. Greater removal by adsorption was observed in long-chain homologues, when compared to short-chain homologues (C13 > C10), whereas the opposite occurred for biodegradation (C10 > C13). The C13 homologue was adsorbed to a great extent (in mass) in T4, T5 and T6, and may also have inhibited methane formation in these treatments. PMID:27088975

  17. TRPM7 channel regulates PDGF-BB-induced proliferation of hepatic stellate cells via PI3K and ERK pathways

    SciTech Connect

    Fang, Ling Zhan, Shuxiang; Huang, Cheng; Cheng, Xi; Lv, Xiongwen; Si, Hongfang; Li, Jun

    2013-11-01

    TRPM7, a non-selective cation channel of the TRP channel superfamily, is implicated in diverse physiological and pathological processes including cell proliferation. Recently, TRPM7 has been reported in hepatic stellate cells (HSCs). Here, we investigated the contribution role of TRPM7 in activated HSC-T6 cell (a rat hepatic stellate cell line) proliferation. TRPM7 mRNA and protein were measured by RT-PCR and Western blot in rat model of liver fibrosis in vivo and PDGF-BB-activated HSC-T6 cells in vitro. Both mRNA and protein of TRPM7 were dramatically increased in CCl{sub 4}-treated rat livers. Stimulation of HSC-T6 cells with PDGF-BB resulted in a time-dependent increase of TRPM7 mRNA and protein. However, PDGF-BB-induced HSC-T6 cell proliferation was inhibited by non-specific TRPM7 blocker 2-aminoethoxydiphenyl borate (2-APB) or synthetic siRNA targeting TRPM7, and this was accompanied by downregulation of cell cycle proteins, cyclin D1, PCNA and CDK4. Blockade of TRPM7 channels also attenuated PDGF-BB induced expression of myofibroblast markers as measured by the induction of α-SMA and Col1α1. Furthermore, the phosphorylation of ERK and AKT, associated with cell proliferation, decreased in TRPM7 deficient HSC-T6 cells. These observations suggested that TRPM7 channels contribute to perpetuated fibroblast activation and proliferation of PDGF-BB induced HSC-T6 cells via the activation of ERK and PI3K pathways. Therefore, TRPM7 may constitute a useful target for the treatment of liver fibrosis. - Highlights: • Upregulation of TRPM7 mRNA and protein in the fibrotic livers from CCl{sub 4}-treated rats. • Increasing expression of TRPM7 mRNA and protein during HSC activation. • Blockade of TRPM7 inhibited the PDGF-BB induced proliferation of HSC-T6 cells. • Blockade of TRPM7 decreased α-SMA and Col1α1 expressions in activated HSC-T6 cells. • TRPM7 up-regulation contributes to the activation of ERK and AKT pathways.

  18. Microstates of a neutral black hole in M theory.

    PubMed

    Emparan, Roberto; Horowitz, Gary T

    2006-10-01

    We consider vacuum solutions in M theory of the form of a five-dimensional Kaluza-Klein black hole cross T6. In a certain limit, these include the five-dimensional neutral rotating black hole (cross T6). From a type-IIA standpoint, these solutions carry D0 and D6 charges. We show that there is a simple D-brane description which precisely reproduces the Hawking-Bekenstein entropy in the extremal limit, even though supersymmetry is completely broken. PMID:17155239

  19. Effect of copper content on corrosion behavior and chromate conversion coating protection of 7xxx series aluminum alloys

    NASA Astrophysics Data System (ADS)

    Meng, Qingjiang

    The addition of Cu in Al-Zn-Mg alloys increases the mechanical strength and resistance to stress corrosion cracking of 7xxx series aluminum alloys (AA7xxx). The peak aged T6 temper provides the maximum mechanical strength by precipitation hardening. However, the presence of noble Cu makes AA7xxx-T6 more susceptible to localized corrosion, such as pitting, crevice and intergranular corrosion (IGC). In order to protect AA7xxx-T6 from localized corrosion, protective chromate conversion coatings (CCCs) must be used. Cu has been reported to affect the CCC protection performance. The exact roles of Cu content in corrosion behavior and CCC protection of AA7xxx-T6 are the focus of this study. Polarization and Electrochemical Impedance Spectroscopy (EIS) approaches were used in combination with materials characterization techniques, such as Focused Ion Beam (FIB), SEM, TEM, High Resolution TEM (HRTEM), Scanning TEM (STEM), and X-ray Photoelectron Spectrometry (XPS). Electrochemical tests on AA7xxx-T6 with various Cu content in deaerated chloride solution found that all alloys except for essentially Cu-free AA7004-T6 had two breakdown potentials, which increased logarithmically with increasing Cu content. Transient dissolution of the fine hardening precipitates and the surrounding solid solution in a thin surface layer was found in the Cu-containing alloys polarized at potentials between the two breakdown potentials. Stable dissolution associated with combined IGC and selective grain attack was found above the second breakdown potential. EIS tests revealed that the overall influence of Cu on the corrosion behavior was detrimental due to Cu enrichment in aerated chloride solution. TEM and STEM analysis revealed that CCC was heterogeneous on the heterogeneous microstructure of AA7075-T6. The coatings formed on coarse intermetallic particles were much thinner than CCC formed on the matrix. It was found that the CCC formed on the matrix mainly consisted of a CrIIIOOH backbone

  20. TGF-β1-elevated TRPM7 channel regulates collagen expression in hepatic stellate cells via TGF-β1/Smad pathway

    SciTech Connect

    Fang, Ling; Huang, Cheng; Meng, Xiaoming; Wu, Baoming; Ma, Taotao; Liu, Xuejiao; Zhu, Qian; Zhan, Shuxiang; Li, Jun

    2014-10-15

    Transdifferentiation of hepatic stellate cells (HSCs) into myofibroblasts plays a critical role in the development of liver fibrosis, since myofibroblasts are the key cells responsible for excessive deposition of ECM proteins. Transient receptor potential melastatin 7 (TRPM7), a non-selective cation channel with protein serine/threonine kinase activity, has been demonstrated to function in the proliferation of activated HSCs. Here, we investigated the functional role of TRPM7 in collagen deposition in activated HSC-T6 cells (a rat hepatic stellate cell line). TRPM7 mRNA and protein were measured by Real-time PCR and Western blot in TGF-β1-activated HSC-T6 cells in vitro. Results demonstrated that TRPM7 protein was dramatically increased in fibrotic human livers. Stimulation of HSC-T6 cells with TGF-β1 increased TRPM7 mRNA and protein level in a time-dependent manner. Nevertheless, TGF-β1-elicited upregulation of TRPM7 in HSC-T6 cells was abrogated by SB431542 (TGF-β1 receptor blocker) or SIS3 (inhibitor of Smad3 phosphorylation). Additionally, blockade of TRPM7 channels with non-specific TRPM7 blocker 2-APB or synthetic siRNA targeting TRPM7 attenuated TGF-β1-induced expression of myofibroblast markers, as measured by the induction of α-SMA and Col1α1. Silencing TRPM7 also increased the ratio of MMPs/TIMPs by increasing MMP-13 expression and decreasing TIMP-1 and TIMP-2 levels. Strikingly, phosphorylation of p-Smad2 and p-Smad3, associated with collagen production, was decreased in TRPM7 deficient HSC-T6 cells. These observations suggested that TGF-β1 elevates TRPM7 expression in HSCs via Smad3-dependant mechanisms, which in turn contributes Smad protein phosphorylation, and subsequently increases fibrous collagen expression. Therefore, TRPM7 may constitute a useful target for the treatment of liver fibrosis. - Highlights: • Upregulation of TRPM7 protein in human fibrotic livers • Upregulation of TRPM7 by TGF-β1 elicited Smad signaling in HSC-T6 cells

  1. An in-situ study in SEM of delamination in several graphite/epoxy composite material systems

    NASA Technical Reports Server (NTRS)

    Bradley, W. L.

    1986-01-01

    A three point bend fixture compatible with the current loading stage for the Scanning Electron Microscope was designed and fabricated. End-notched flexure tests were run on several materials. Work to date was on AS4/3502, T6T145/F155, and T6T145/F185. Fracture toughness was measured. Fracture of neat resin specimens was begun. The decrease in delaminatin fracture toughness, compared to neat resin toughness, due to rigid fiber filler and interlaminar failure is addressed. An experimental program was designed to try to determine the proper interpretatin for apparent microcracking in neat resin specimens.

  2. Intraspecies Competition in Serratia marcescens Is Mediated by Type VI-Secreted Rhs Effectors and a Conserved Effector-Associated Accessory Protein

    PubMed Central

    Alcoforado Diniz, Juliana

    2015-01-01

    ABSTRACT The type VI secretion system (T6SS) is widespread in Gram-negative bacteria and can deliver toxic effector proteins into eukaryotic cells or competitor bacteria. Antibacterial T6SSs are increasingly recognized as key mediators of interbacterial competition and may contribute to the outcome of many polymicrobial infections. Multiple antibacterial effectors can be delivered by these systems, with diverse activities against target cells and distinct modes of secretion. Polymorphic toxins containing Rhs repeat domains represent a recently identified and as-yet poorly characterized class of T6SS-dependent effectors. Previous work had revealed that the potent antibacterial T6SS of the opportunistic pathogen Serratia marcescens promotes intraspecies as well as interspecies competition (S. L. Murdoch, K. Trunk, G. English, M. J. Fritsch, E. Pourkarimi, and S. J. Coulthurst, J Bacteriol 193:6057–6069, 2011, http://dx.doi.org/10.1128/JB.05671-11). In this study, two new Rhs family antibacterial effectors delivered by this T6SS have been identified. One of these was shown to act as a DNase toxin, while the other contains a novel, cytoplasmic-acting toxin domain. Importantly, using S. marcescens, it has been demonstrated for the first time that Rhs proteins, rather than other T6SS-secreted effectors, can be the primary determinant of intraspecies competition. Furthermore, a new family of accessory proteins associated with T6SS effectors has been identified, exemplified by S. marcescens EagR1, which is specifically required for deployment of its associated Rhs effector. Together, these findings provide new insight into how bacteria can use the T6SS to deploy Rhs-family effectors and mediate different types of interbacterial interactions. IMPORTANCE Infectious diseases caused by bacterial pathogens represent a continuing threat to health and economic prosperity. To counter this threat, we must understand how such organisms survive and prosper. The type VI secretion

  3. The optimized mechanical properties of the new aluminum alloy AA 6069

    SciTech Connect

    Bergsma, S.C.; Kassner, M.E.; Li, X.; Delos-Reyes, M.A.; Hayes, T.A.

    1996-02-01

    AA 6069, a new aluminum alloy, has been developed for application in hot and cold extrusion and forging. It contains {approximately}2 Mg + Si, {approximately}1% Cu, 0.2% Cr, and 0.1% V. Nominal T6 properties of the ingot without hot or cold deformation are 415 MPa (60 ksi) ultimate tensile strength (UTS), 380 MPa (55 ksi) yield strength, and 12% elongation. Properties after hot and cold extrusion in the T6 condition rate from 380 to 490 MPa (55 to 71 ksi) UTS, 345 to 450 MPa (50 to 65 ksi) yield strength, and 10 to 22% elongation. This alloy also has favorable fatigue and corrosion-fatigue properties due to a combination of composition, high solidification rate, controlled homogenization, thermal and mechanical processing, and T6 practice. Current development applications include cold-impact air-bag components, high-pressure cylinders, and automotive suspension and drive-train parts. Unlike alloys 2024-T3 and 7129-T6, of comparable strength, diluted 6069 is scrap compatible with many other 5xxx and 6xxx alloys.

  4. The optimized mechanical properties of the new aluminum alloy AA 6069

    NASA Astrophysics Data System (ADS)

    Bergsma, S. C.; Kassner, M. E.; Li, X.; Delos-Reyes, U. A.; Hayes, T. A.

    1996-02-01

    AA 6069, a new aluminum alloy, has been developed for application in hot and cold extrusion and forging. It contains ~2Mg + Si, ~1% Cu, 0.2% Cr, and 0.1% V. Nominal T6 properties of the ingot without hot or cold deformation are 415 MPa (60 ksi) ultimate tensile strength (UTS), 380 MPa (55 ksi) yield strength, and 12 % elongation. Properties after hot and cold extrusion in the T6 condition range from 380 to 490 MPa (55 to 71 ksi) UTS, 345 to 450 MPa (50 to 65 ksi) yield strength, and 10 to 22 % elongation. This alloy also has favorable fatigue and corrosion- fatigue properties due to a combination of composition, high solidification rate, controlled homogenization, thermal and mechanical processing, and T6 practice. Current developmental applications include cold- impact air- bag components, high- pressure cylinders, and automotive suspension and drive- train parts. Unlike alloys 2024- T3 and 7129- T6, of comparable strength, diluted 6069 is scrap compatible with many other 5xxx and 6xxx alloys.

  5. Plumbagin Ameliorates CCl4-Induced Hepatic Fibrosis in Rats via the Epidermal Growth Factor Receptor Signaling Pathway

    PubMed Central

    Chen, Si; Chen, Yi; Chen, Bi; Cai, Yi-jing; Zou, Zhuo-lin; Wang, Jin-guo; Lin, Zhuo; Wang, Xiao-dong; Fu, Li-yun; Hu, Yao-ren; Chen, Yong-ping; Chen, Da-zhi

    2015-01-01

    Epidermal growth factor (EGF) and its signaling molecules, EGFreceptor (EGFR) and signal transducer and activator of transcription factor 3 (STAT3), have been considered to play a role in liver fibrosis and cirrhosis. Plumbagin (PL) is an extracted component from the plant and has been used to treat different kinds of cancer. However, its role in regulation of EGFR and STAT3 during liver fibrosis has not been investigated. In this study, the effects of PL on the regulation of EGFR and STAT3 were investigated in carbon tetrachloride (CCl4) induced liver fibrosis and hepatic stellate cells (HSC-T6). PL significantly attenuated liver injury and fibrosis in CCl4 treated rats. At concentrations of 2 to 6 μM, PL did not induce significant cytotoxicity of HSC-T6 cells. Moreover, PL reduced phosphorylation of EGFR and STAT3 in both fibrotic liver and heparin-binding EGF-like growth factor (HB-EGF) treated HSC-T6 cells. Furthermore, PL reduced the expression of α-SMA, EGFR, and STAT3 in both fibrotic liver and HB-EGF treated HSC-T6 cells. In conclusion, plumbagin could ameliorate the development of hepatic fibrosis through its downregulation of EGFR and STAT3 in the liver, especially in hepatic stellate cells. PMID:26550019

  6. 27 CFR 9.195 - Alta Mesa.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... County, California, and is entirely within the Lodi viticultural area (27 CFR 9.107). The Alta Mesa... its intersection with Dillard Road, section 6, T6N, R7E (Elk Grove Quadrangle); then (5) Proceed northeast 2.6 miles on Dillard Road to its intersection with Lee Shorthorn Road, T7N, R7E...

  7. 27 CFR 9.195 - Alta Mesa.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... County, California, and is entirely within the Lodi viticultural area (27 CFR 9.107). The Alta Mesa... its intersection with Dillard Road, section 6, T6N, R7E (Elk Grove Quadrangle); then (5) Proceed northeast 2.6 miles on Dillard Road to its intersection with Lee Shorthorn Road, T7N, R7E...

  8. 27 CFR 9.195 - Alta Mesa.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... County, California, and is entirely within the Lodi viticultural area (27 CFR 9.107). The Alta Mesa... its intersection with Dillard Road, section 6, T6N, R7E (Elk Grove Quadrangle); then (5) Proceed northeast 2.6 miles on Dillard Road to its intersection with Lee Shorthorn Road, T7N, R7E...

  9. 27 CFR 9.195 - Alta Mesa.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... County, California, and is entirely within the Lodi viticultural area (27 CFR 9.107). The Alta Mesa... its intersection with Dillard Road, section 6, T6N, R7E (Elk Grove Quadrangle); then (5) Proceed northeast 2.6 miles on Dillard Road to its intersection with Lee Shorthorn Road, T7N, R7E...

  10. 27 CFR 9.195 - Alta Mesa.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... County, California, and is entirely within the Lodi viticultural area (27 CFR 9.107). The Alta Mesa... its intersection with Dillard Road, section 6, T6N, R7E (Elk Grove Quadrangle); then (5) Proceed northeast 2.6 miles on Dillard Road to its intersection with Lee Shorthorn Road, T7N, R7E...

  11. 27 CFR 9.206 - Shawnee Hills.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...) Approved maps. The United States Geological Survey (USGS) 1:250,000-scale topographic map used to determine... between the Ohio and Mississippi Rivers, and largely within the Shawnee National Forest. The boundary of... intersection with the surveyed boundary line between Township 6 South (T6S) and Township 7 South (T7S); then...

  12. 27 CFR 9.206 - Shawnee Hills.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) Approved maps. The United States Geological Survey (USGS) 1:250,000-scale topographic map used to determine... between the Ohio and Mississippi Rivers, and largely within the Shawnee National Forest. The boundary of... intersection with the surveyed boundary line between Township 6 South (T6S) and Township 7 South (T7S); then...

  13. 27 CFR 9.206 - Shawnee Hills.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) Approved maps. The United States Geological Survey (USGS) 1:250,000-scale topographic map used to determine... between the Ohio and Mississippi Rivers, and largely within the Shawnee National Forest. The boundary of... intersection with the surveyed boundary line between Township 6 South (T6S) and Township 7 South (T7S); then...

  14. 27 CFR 9.206 - Shawnee Hills.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...) Approved maps. The United States Geological Survey (USGS) 1:250,000-scale topographic map used to determine... between the Ohio and Mississippi Rivers, and largely within the Shawnee National Forest. The boundary of... intersection with the surveyed boundary line between Township 6 South (T6S) and Township 7 South (T7S); then...

  15. Galvanic Corrosion In (Graphite/Epoxy)/Alloy Couples

    NASA Technical Reports Server (NTRS)

    Danford, Merlin D.; Higgins, Ralph H.

    1988-01-01

    Effects of galvanic coupling between graphite/epoxy composite material, G/E, and D6AC steel, 6061-T6 aluminum, and Inconel(R) 718 nickel alloy in salt water described in report. Introductory section summarizes previous corrosion studies of G/E with other alloys. Details of sample preparation presented along with photographs of samples before and after immersion.

  16. Dioxygenases catalyze O-demethylation and O,O-demethylenation with widespread roles in benzylisoquinoline alkaloid metabolism in opium poppy.

    PubMed

    Farrow, Scott C; Facchini, Peter J

    2013-10-01

    In opium poppy, the antepenultimate and final steps in morphine biosynthesis are catalyzed by the 2-oxoglutarate/Fe(II)-dependent dioxygenases, thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM). Further investigation into the biochemical functions of CODM and T6ODM revealed extensive and unexpected roles for such enzymes in the metabolism of protopine, benzo[c]phenanthridine, and rhoeadine alkaloids. When assayed with a wide range of benzylisoquinoline alkaloids, CODM, T6ODM, and the functionally unassigned paralog DIOX2, renamed protopine O-dealkylase, showed novel and efficient dealkylation activities, including regio- and substrate-specific O-demethylation and O,O-demethylenation. Enzymes catalyzing O,O-demethylenation, which cleave a methylenedioxy bridge leaving two hydroxyl groups, have previously not been reported in plants. Similar cleavage of methylenedioxy bridges on substituted amphetamines is catalyzed by heme-dependent cytochromes P450 in mammals. Preferred substrates for O,O-demethylenation by CODM and protopine O-dealkylase were protopine alkaloids that serve as intermediates in the biosynthesis of benzo[c]phenanthridine and rhoeadine derivatives. Virus-induced gene silencing used to suppress the abundance of CODM and/or T6ODM transcripts indicated a direct physiological role for these enzymes in the metabolism of protopine alkaloids, and they revealed their indirect involvement in the formation of the antimicrobial benzo[c]phenanthridine sanguinarine and certain rhoeadine alkaloids in opium poppy.

  17. Diversity of Clinical and Environmental Isolates of Vibrio cholerae in Natural Transformation and Contact-Dependent Bacterial Killing Indicative of Type VI Secretion System Activity.

    PubMed

    Bernardy, Eryn E; Turnsek, Maryann A; Wilson, Sarah K; Tarr, Cheryl L; Hammer, Brian K

    2016-05-01

    The bacterial pathogen Vibrio cholerae can occupy both the human gut and aquatic reservoirs, where it may colonize chitinous surfaces that induce the expression of factors for three phenotypes: chitin utilization, DNA uptake by natural transformation, and contact-dependent bacterial killing via a type VI secretion system (T6SS). In this study, we surveyed a diverse set of 53 isolates from different geographic locales collected over the past century from human clinical and environmental specimens for each phenotype outlined above. The set included pandemic isolates of serogroup O1, as well as several serogroup O139 and non-O1/non-O139 strains. We found that while chitin utilization was common, only 22.6% of the isolates tested were proficient at chitin-induced natural transformation, suggesting that transformation is expendable. Constitutive contact-dependent killing of Escherichia coli prey, which is indicative of a functional T6SS, was rare among clinical isolates (only 4 of 29) but common among environmental isolates (22 of 24). These results bolster the pathoadaptive model in which tight regulation of T6SS-mediated bacterial killing is beneficial in a human host, whereas constitutive killing by environmental isolates may give a competitive advantage in natural settings. Future sequence analysis of this set of diverse isolates may identify previously unknown regulators and structural components for both natural transformation and T6SS. PMID:26944842

  18. Roles of Hcp family proteins in the pathogenesis of the porcine extraintestinal pathogenic Escherichia coli type VI secretion system

    PubMed Central

    Peng, Ying; Wang, Xiangru; Shou, Jin; Zong, Bingbing; Zhang, Yanyan; Tan, Jia; Chen, Jing; Hu, Linlin; Zhu, Yongwei; Chen, Huanchun; Tan, Chen

    2016-01-01

    Hcp (hemolysin-coregulated protein) is considered a vital component of the functional T6SS (Type VI Secretion System), which is a newly discovered secretion system. Our laboratory has previously sequenced the whole genome of porcine extraintestinal pathogenic E. coli (ExPEC) strain PCN033, and identified an integrated T6SS encoding three different hcp family genes. In this study, we first identified a functional T6SS in porcine ExPEC strain PCN033, and demonstrated that the Hcp family proteins were involved in bacterial competition and the interactions with other cells. Interestingly, the three Hcp proteins had different functions. Hcp2 functioned predominantly in bacterial competition; all three proteins were involved in the colonization of mice; and Hcp1 and Hcp3 were predominantly contributed to bacterial-eukaryotic cell interactions. We showed an active T6SS in porcine ExPEC strain PCN033, and the Hcp family proteins had different functions in their interaction with other bacteria or host cells. PMID:27229766

  19. Black holes as D3-branes on Calabi-Yau threefolds

    NASA Astrophysics Data System (ADS)

    Bertolini, Matteo; Frè, Pietro; Iengo, Roberto; Scrucca, Claudio A.

    1998-07-01

    We show how an extremal Reissner-Nordström black hole can be obtained by wrapping a dyonic D3-brane on a Calabi-Yau manifold. In the orbifold limit T6/Z3, we explicitly show the correspondence between the solution of the supergravity equations of motion and the D-brane boundary state description of such a black hole.

  20. 50 CFR Appendix I to Part 37 - Legal Description of the Coastal Plain, Arctic National Wildlife Refuge, Alaska

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... mean high tide of the Beaufort Lagoon, located in section 28, T. 6 N., R. 40 E., Umiat Meridian; Thence... Meridian at the line of extreme low tide; Thence northwesterly, along the northerly boundary of the Arctic National Wildlife Refuge at the line of extreme low tide on the seaward side of all offshore bars,...

  1. 50 CFR Appendix I to Part 37 - Legal Description of the Coastal Plain, Arctic National Wildlife Refuge, Alaska

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... mean high tide of the Beaufort Lagoon, located in section 28, T. 6 N., R. 40 E., Umiat Meridian; Thence... Meridian at the line of extreme low tide; Thence northwesterly, along the northerly boundary of the Arctic National Wildlife Refuge at the line of extreme low tide on the seaward side of all offshore bars,...

  2. 50 CFR Appendix I to Part 37 - Legal Description of the Coastal Plain, Arctic National Wildlife Refuge, Alaska

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... mean high tide of the Beaufort Lagoon, located in section 28, T. 6 N., R. 40 E., Umiat Meridian; Thence... Meridian at the line of extreme low tide; Thence northwesterly, along the northerly boundary of the Arctic National Wildlife Refuge at the line of extreme low tide on the seaward side of all offshore bars,...

  3. 14 CFR 91.1507 - Fuel tank system inspection program.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...) Illyushin Aviation IL 96T (6) Bristol Aircraft Britannia 305 (7) Handley Page Herald Type 300 (8) Avions... or § 25.1529 and part 25, Appendix H, of this chapter, in effect on June 6, 2001 (including those..., or under § 25.1529 in effect on June 6, 2001, the operator must include in the inspection program...

  4. Ex Vivo Smooth Muscle Pharmacological Effects of a Novel Bradykinin-Related Peptide, and Its Analogue, from Chinese Large Odorous Frog, Odorrana livida Skin Secretions

    PubMed Central

    Xiang, Jie; Wang, Hui; Ma, Chengbang; Zhou, Mei; Wu, Yuxin; Wang, Lei; Guo, Shaodong; Chen, Tianbao; Shaw, Chris

    2016-01-01

    Bradykinin-related peptides (BRPs) are one of the most extensively studied frog secretions-derived peptide families identified from many amphibian species. The diverse primary structures of BRPs have been proven essential for providing valuable information in understanding basic mechanisms associated with drug modification. Here, we isolated, identified and characterized a dodeca-BRP (RAP-L1, T6-BK), with primary structure RAPLPPGFTPFR, from the skin secretions of Chinese large odorous frogs, Odorrana livida. This novel peptide exhibited a dose-dependent contractile property on rat bladder and rat ileum, and increased the contraction frequency on rat uterus ex vivo smooth muscle preparations; it also showed vasorelaxant activity on rat tail artery smooth muscle. In addition, the analogue RAP-L1, T6, L8-BK completely abolished these effects on selected rat smooth muscle tissues, whilst it showed inhibition effect on bradykinin-induced rat tail artery relaxation. By using canonical antagonist for bradykinin B1 or B2 type receptors, we found that RAP-L1, T6-BK -induced relaxation of the arterial smooth muscle was very likely to be modulated by B2 receptors. The analogue RAP-L1, T6, L8-BK further enhanced the bradykinin inhibitory activity only under the condition of co-administration with HOE140 on rat tail artery, suggesting a synergistic inhibition mechanism by which targeting B2 type receptors. PMID:27690099

  5. The Pseudomonas aeruginosa Type VI Secretion PGAP1-like Effector Induces Host Autophagy by Activating Endoplasmic Reticulum Stress.

    PubMed

    Jiang, Feng; Wang, Xia; Wang, Bei; Chen, Lihong; Zhao, Zhendong; Waterfield, Nicholas R; Yang, Guowei; Jin, Qi

    2016-08-01

    Pseudomonas aeruginosa is an opportunistic pathogen that regularly causes nosocomial infections in hospitalized patients. The type VI secretion system (T6SS) is responsible for the secretion of numerous virulence effector proteins that can both interfere with competing microbes and manipulate host cells. Here, we report a detailed investigation of a P. aeruginosa H2-T6SS-dependent phospholipase effector, TplE, which acts as a trans-kingdom toxin. Delivery of TplE to the periplasmic space of rival bacteria leads to growth inhibition. Importantly, TplE, also contains a eukaryotic PGAP1-like domain, which targets the host ER apparatus, ultimately leading to disruption of the ER. TplE activity leads to the activation of the unfolded protein response (UPR) through the IRE1α-XBP1 pathway, enhancing autophagic flux. These findings indicate that this T6SS-delivered phospholipase effector is active against both prokaryotic and eukaryotic cellular targets, highlighting the T6SS as a versatile weapon in the Pseudomonas arsenal. PMID:27477276

  6. School Size, Cost and Quality

    ERIC Educational Resources Information Center

    Ratsoy, Eugene W.; Bumbarger, Chester S.

    1976-01-01

    Despite the trend toward consolidation of schools, many small schools continue to exist. The challenge is to identify and implement the changes that will improve these schools. (Available from Canadian Administrator Business Manager, Department of Educational Administration, The University of Alberta, Edmonton, Alberta, Canada T6G 2G5; $0.50…

  7. Educational Vouchers

    ERIC Educational Resources Information Center

    Pickard, Brent W.; Richards, Donald M.

    1976-01-01

    Summarizes the rationale for educational voucher systems, outlines model voucher systems, discusses implications of voucher systems, and suggests a generalized plan for applying voucher systems in continuing education. (Available from the Department of Educational Administration, The University of Alberta, Edmonton, Alberta, Canada, T6G 2G5;…

  8. Professional Role Orientation of Women Administrators and Women Teachers

    ERIC Educational Resources Information Center

    Nixon, Mary; Gue, L. R.

    1975-01-01

    The professional role orientation of women teachers and women administrators is compared with a limited number of variables. (Available from the Department of Educational Administration, The University of Alberta, Edmonton, Canada T6G 2G5; $4.00 annually.) (MLF)

  9. Administering the Open-Area Elementary School

    ERIC Educational Resources Information Center

    Small, J. M.

    1975-01-01

    Examines the outcome of a study of the operation of open area schools in the Separate School System of Edmonton. (Available from C. A. Business Manager, Department of Educational Administration, The University of Alberta, Edmonton, Alberta, Canada T6G 2G5; $0.50, single copy.) (Author/IRT)

  10. Indian Government and Indian Education.

    ERIC Educational Resources Information Center

    Starblanket, Noel V.

    1981-01-01

    Accountability for Indian education must be shared among the chiefs and their councils, the Indian leaders at all levels, parents and students. This may be accomplished by Indian control of Indian education. Available from: Department of Educational Foundations, 5-109 Education North, University of Alberta, Edmonton, AB, Canada, T6G 2G5. (ERB)

  11. The Role of the Native Woman in a Native Society.

    ERIC Educational Resources Information Center

    Swampy, Grace Marie

    1982-01-01

    Eighteen Plains Cree-speaking women, aged 15-93, were interviewed to determine the persistence of traditional beliefs as they affect the role of women. "The major implication is that the culture is alive." Available: Department of Educational Foundations, 5-109 Education North, University of Alberta, Edmonton, Alberta, Canada, T6G 2G5. (LC)

  12. Militancy and Accommodativeness in Teachers' Negotiations: Two Ontario Surveys

    ERIC Educational Resources Information Center

    Fris, J.

    1976-01-01

    Reports findings of two surveys of Ontario elementary and secondary teachers that measured teachers' attitudes regarding collective bargaining tactics and classified teachers' responses according to their militancy or accomodativeness. Available from Department of Educational Administration, The University of Alberta, Edmonton, Alberta, Canada T6G…

  13. Teachers' Aides: Tasks and Concerns

    ERIC Educational Resources Information Center

    Balderson, James H.; Nixon, Mary

    1976-01-01

    Addresses three questions: (1) What tasks do aides perform? (2) Does training make a difference in the type of tasks aides perform? (3) What are the concerns of aides? (Available from the Department of Educational Administration, The University of Alberta, Edmonton, Alberta, Canada T6G 2G5; $0.50, single copy.) (Author/IRT)

  14. Fractographic analysis of the low energy fracture of an aluminum alloy

    NASA Technical Reports Server (NTRS)

    Tanaka, J.; Pampillo, C. A.; Low, J. R., Jr.

    1972-01-01

    A study of the fracture process in a high strength aluminum alloy, 2014T6, was undertaken to identify the void nucleating particles in this material, to determine their composition, and to suggest means by which they might be eliminated without loss of strength.

  15. Dioxygenases Catalyze O-Demethylation and O,O-Demethylenation with Widespread Roles in Benzylisoquinoline Alkaloid Metabolism in Opium Poppy*

    PubMed Central

    Farrow, Scott C.; Facchini, Peter J.

    2013-01-01

    In opium poppy, the antepenultimate and final steps in morphine biosynthesis are catalyzed by the 2-oxoglutarate/Fe(II)-dependent dioxygenases, thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM). Further investigation into the biochemical functions of CODM and T6ODM revealed extensive and unexpected roles for such enzymes in the metabolism of protopine, benzo[c]phenanthridine, and rhoeadine alkaloids. When assayed with a wide range of benzylisoquinoline alkaloids, CODM, T6ODM, and the functionally unassigned paralog DIOX2, renamed protopine O-dealkylase, showed novel and efficient dealkylation activities, including regio- and substrate-specific O-demethylation and O,O-demethylenation. Enzymes catalyzing O,O-demethylenation, which cleave a methylenedioxy bridge leaving two hydroxyl groups, have previously not been reported in plants. Similar cleavage of methylenedioxy bridges on substituted amphetamines is catalyzed by heme-dependent cytochromes P450 in mammals. Preferred substrates for O,O-demethylenation by CODM and protopine O-dealkylase were protopine alkaloids that serve as intermediates in the biosynthesis of benzo[c]phenanthridine and rhoeadine derivatives. Virus-induced gene silencing used to suppress the abundance of CODM and/or T6ODM transcripts indicated a direct physiological role for these enzymes in the metabolism of protopine alkaloids, and they revealed their indirect involvement in the formation of the antimicrobial benzo[c]phenanthridine sanguinarine and certain rhoeadine alkaloids in opium poppy. PMID:23928311

  16. Evaluating of NASA-Langley Research Center explosion seam welding

    NASA Technical Reports Server (NTRS)

    Otto, H. E.; Wittman, R.

    1977-01-01

    An explosion bonding technique to meet current fabrication requirements was demonstrated. A test program was conducted on explosion bonded joints, compared to fusion joints in 6061-T6 aluminum. The comparison was made in required fixtures, non-destructive testing, static strength and fatigue strength.

  17. Synergistic derepression of gibberellin signaling by removing RGA and GAI function in Arabidopsis thaliana.

    PubMed

    Dill, A; Sun, T

    2001-10-01

    RGA and GAI are negative regulators of the gibberellin (GA) signal transduction pathway in Arabidopsis thaliana. These genes may have partially redundant functions because they are highly homologous, and plants containing single null mutations at these loci are phenotypically similar to wild type. Previously, rga loss-of-function mutations were shown to partially suppress defects of the GA-deficient ga1-3 mutant. Phenotypes rescued include abaxial trichome initiation, rosette radius, flowering time, stem elongation, and apical dominance. Here we present work showing that the rga-24 and gai-t6 null mutations have a synergistic effect on plant growth. Although gai-t6 alone has little effect, when combined with rga-24, they completely rescued the above defects of ga1-3 to wild-type or GA-overdose phenotype. However, seed germination and flower development defects were not restored. Additionally, rga-24 and rga-24/gai-t6 but not gai-t6 alone caused increased feedback inhibition of expression of a GA biosynthetic gene in both the ga1-3 and wild-type backgrounds. These results demonstrate that RGA and GAI have partially redundant functions in maintaining the repressive state of the GA-signaling pathway, but RGA plays a more dominant role than GAI. Removing both RGA and GAI function allows for complete derepression of many aspects of GA signaling.

  18. Hcp2, a Secreted Protein of the Phytopathogen Pseudomonas syringae pv. Tomato DC3000, Is Required for Fitness for Competition against Bacteria and Yeasts

    PubMed Central

    Mosorin, Hanna; Dorati, Federico; Wu, Ru-Fen; Roine, Elina; Taira, Suvi; Nissinen, Riitta; Mattinen, Laura; Jackson, Robert; Pirhonen, Minna

    2012-01-01

    When analyzing the secretome of the plant pathogen Pseudomonas syringae pv. tomato DC3000, we identified hemolysin-coregulated protein (Hcp) as one of the secreted proteins. Hcp is assumed to be an extracellular component of the type VI secretion system (T6SS). Two copies of hcp genes are present in the P. syringae pv. tomato DC3000 genome, hcp1 (PSPTO_2539) and hcp2 (PSPTO_5435). We studied the expression patterns of the hcp genes and tested the fitness of hcp knockout mutants in host plant colonization and in intermicrobial competition. We found that the hcp2 gene is expressed most actively at the stationary growth phase and that the Hcp2 protein is secreted via the T6SS and appears in the culture medium as covalently linked dimers. Expression of hcp2 is not induced in planta and does not contribute to virulence in or colonization of tomato or Arabidopsis plants. Instead, hcp2 is required for survival in competition with enterobacteria and yeasts, and its function is associated with the suppression of the growth of these competitors. This is the first report on bacterial T6SS-associated genes functioning in competition with yeast. Our results suggest that the T6SS of P. syringae may play an important role in bacterial fitness, allowing this plant pathogen to survive under conditions where it has to compete with other microorganisms for resources. PMID:22753062

  19. Diversity of Clinical and Environmental Isolates of Vibrio cholerae in Natural Transformation and Contact-Dependent Bacterial Killing Indicative of Type VI Secretion System Activity

    PubMed Central

    Bernardy, Eryn E.; Turnsek, Maryann A.; Wilson, Sarah K.; Tarr, Cheryl L.

    2016-01-01

    The bacterial pathogen Vibrio cholerae can occupy both the human gut and aquatic reservoirs, where it may colonize chitinous surfaces that induce the expression of factors for three phenotypes: chitin utilization, DNA uptake by natural transformation, and contact-dependent bacterial killing via a type VI secretion system (T6SS). In this study, we surveyed a diverse set of 53 isolates from different geographic locales collected over the past century from human clinical and environmental specimens for each phenotype outlined above. The set included pandemic isolates of serogroup O1, as well as several serogroup O139 and non-O1/non-O139 strains. We found that while chitin utilization was common, only 22.6% of the isolates tested were proficient at chitin-induced natural transformation, suggesting that transformation is expendable. Constitutive contact-dependent killing of Escherichia coli prey, which is indicative of a functional T6SS, was rare among clinical isolates (only 4 of 29) but common among environmental isolates (22 of 24). These results bolster the pathoadaptive model in which tight regulation of T6SS-mediated bacterial killing is beneficial in a human host, whereas constitutive killing by environmental isolates may give a competitive advantage in natural settings. Future sequence analysis of this set of diverse isolates may identify previously unknown regulators and structural components for both natural transformation and T6SS. PMID:26944842

  20. Co-culture of mouse embryos with oviduct and uterine cells prepared from mice at different days of pseudopregnancy.

    PubMed

    Sakkas, D; Trounson, A O

    1990-09-01

    Oviduct and uterine cell cultures were prepared from mice at different days of pseudopregnancy and their effects on the development of 1- and 8-cell mouse embryos in co-culture were examined. One-cell mouse embryos in co-culture with oviduct cells from 20 h to 120 h after hCG had a mean (+/- s.e.) cell number of 70.1 +/- 3.6, significantly (P less than 0.001) higher compared with those cultured in Whittingham's T6 medium supplemented with 5% fetal calf serum (T6 + 5% FCS) (30.4 +/- 1.6). Transfer of embryos, at 96 h after hCG, to synchronous pseudopregnant recipients showed that more embryos in oviduct co-culture formed fetuses than those cultured in T6 + 5% FCS. Co-culture of 1-cell embryos with uterine cells did not confer an advantage in cell numbers over T6 + 5% FCS. However, more 8-cell embryos formed blastocyst outgrowths after 100 h in co-culture with uterine cells prepared from mice at Day 3 of pseudopregnancy than with uterine cultures prepared from mice at Day 1 of pseudopregnancy or oviduct cells. In addition, there was further improvement when the Day 3 uterine co-cultures were supplemented with 1 or 10 ng progesterone/ml. These results highlight the importance of the oviduct and uterine cells during the different stages of preimplantation embryo development.

  1. A highly sensitive and selective immunoassay for the detection of tetrabromobisphenol A in soil and sediment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant. A sensitive and selective enzyme-linked immunosorbent assay (ELISA) for the detection of TBBPA was developed. Six haptens (T1-T6) mimicking different structural elements of TBBPA were synthesized and coupled to keyhole...

  2. [Posttraumatic syringomyelia developing soon after spinal cord injury. A case report].

    PubMed

    Yoshimura, Kazuhiro; Sasaki, Manabu; Morris, Shane; Abekura, Makoto; Yoshimine, Toshiki

    2008-10-01

    We report a case of posttraumatic syringomyelia (PTS) that developed 10 months after spinal cord injury (SCI), A 46-year-old man was involved in a motorcycle accident, in which he received a severe spinal cord injury due to a burst fracture at the T6 level. The patient underwent posterior fixation without decompression at another hospital, and was transferred to our hospital for rehabilitation. Ten months after the SCI, he complained of back and neck pain caused by bending his head backward. MRI showed syringomyelia shaped like a cone extending from the T6 to the C6 level, enlarged by cerebrospinal fluid (CSF) flow toward the syringomyelia at the T6 level. Pain was relieved by syringosubarachnoid shunt implantation and the syrinx disappeared after the operation. PTS emerging 10 months after spinal cord injury is relatively rare. From the radiological and operative findings, PTS was enlarged by the CSF flow, which was hindered at the T6 level by compression of the vertebral body to the spinal cord. In cases of SCI in patients who undergo posterior fixation with insufficient decompression, close attention to PTS is required in the postoperative follow-up. PMID:18975566

  3. Type Six Secretion System of Bordetella bronchiseptica and Adaptive Immune Components Limit Intracellular Survival During Infection.

    PubMed

    Bendor, Liron; Weyrich, Laura S; Linz, Bodo; Rolin, Olivier Y; Taylor, Dawn L; Goodfield, Laura L; Smallridge, William E; Kennett, Mary J; Harvill, Eric T

    2015-01-01

    The Type Six Secretion System (T6SS) is required for Bordetella bronchiseptica cytotoxicity, cytokine modulation, infection, and persistence. However, one-third of recently sequenced Bordetella bronchiseptica strains of the predominantly human-associated Complex IV have lost their T6SS through gene deletion or degradation. Since most human B. bronchiseptica infections occur in immunocompromised patients, we determine here whether loss of Type Six Secretion is beneficial to B. bronchiseptica during infection of immunocompromised mice. Infection of mice lacking adaptive immunity (Rag1-/- mice) with a T6SS-deficient mutant results in a hypervirulent phenotype that is characterized by high numbers of intracellular bacteria in systemic organs. In contrast, wild-type B. bronchiseptica kill their eukaryotic cellular hosts via a T6SS-dependent mechanism that prevents survival in systemic organs. High numbers of intracellular bacteria recovered from immunodeficient mice but only low numbers from wild-type mice demonstrates that B. bronchiseptica survival in an intracellular niche is limited by B and T cell responses. Understanding the nature of intracellular survival during infection, and its effects on the generation and function of the host immune response, are important to contain and control the spread of Bordetella-caused disease. PMID:26485303

  4. Mutation in the S-ribosylhomocysteinase (luxS) gene involved in quorum sensing affects biofilm formation and virulence in a clinical isolate of Aeromonas hydrophila

    EPA Science Inventory

    A diarrheal isolate SSU of Aeromonas hydrophila produces a cytotoxic enterotoxin (Act) with cytotoxic, enterotoxic, and hemolytic activities. Our laboratory has characterized from the above Aeromonas strain, in addition to Act, the type 3- and T6-secretion systems and their effec...

  5. Tests Of Polyurethane And Dichromate Coats On Aluminum

    NASA Technical Reports Server (NTRS)

    Danford, M. D.

    1992-01-01

    Report describes experiments to determine relative effectiveness of new polyurethane and more-conventional dichromate coat in helping to retard corrosion of anodized 6061-T6 aluminum. Concludes by suggesting greater protection against corrosion achieved by combining polyurethane-sealing method with hard-anodizing method and by increasing thickness of coat.

  6. Biotechnology Process Engineering Center at MIT Home

    Science.gov Websites

    ... o PICTo &3;&24;&2;d &17;&2; &1;, &1;, &1; &4;MSWD &30; &1; , &18;&7;&15;Times New Roman &3;&7; 2 . &4; (&1;Z&1;w&1;< &6; &1; - &8; &22; Q )&28;&1;h &6; - &8; &22; )&25;&1;t &6; &2; - &8; &1; &22; ...

  7. Type VI Secretion: not just for pathogenesis anymore

    PubMed Central

    Jani, Andrea J.; Cotter, Peggy A.

    2010-01-01

    Summary Type VI Secretion Systems (T6SS) have been studied primarily in the context of pathogenic bacterial-host interactions. Recent data suggest, however, that these versatile secretion systems may also function to promote commensal or mutualistic relationships between bacteria and eukaryotes, or to mediate cooperative or competitive interactions between bacteria. PMID:20638635

  8. A new fauna from the Colorado group of southern Montana

    USGS Publications Warehouse

    Reeside, John B.

    1925-01-01

    This paper describes a small but interesting fauna collected in 1921 by W. T. Thorn, Jr., Gail F. Moulton, T. W. Stanton, and K. C. Heald in the Crow Indian Reservation in southern Montana. The locality is in sec. 36, T. 6 S., R. 32 E., Big Horn County, and is 2 miles east of the Soap Creek oil field.

  9. The feasibility of producing aluminum-lithium structures for cryogenic tankage applications by laser beam welding

    NASA Technical Reports Server (NTRS)

    Martukanitz, R. P.; Lysher, K. G.

    1993-01-01

    Aluminum-lithium alloys exhibit high strength, high elastic modulus, and low density as well as excellent cryogenic mechanical properties making them ideal material candidates for cryogenic tanks. NASA has proposed the use of 'built-up' structure for panels fabricated into cryogenic tanks replacing current conventional machining. Superplastically formed stiffeners would be joined to sheet (tank skin) that had been roll formed to the radius of the tank in order to produce panels. Aluminum-lithium alloys of interest for producing the built-up structure include alloy 2095-T6 stiffeners to 2095-T8 sheet and alloy 8090-T6 stiffeners to 2090-T83 sheet. Laser welding, with comparable joint properties, offers the following advantages over conventional welding: higher production rates, minimal degradation within the heat affected zones, and full process automation. This study established process parameters for laser beam welding, mechanical property determinations, metallographic characterization, and fabrication of prototype panels. Tensile tests representing partial penetration of the skin alloys provided joint efficiencies between 65 and 77 percent, depending upon alloy and degree of penetration. Results of tension shear tests of lap welds indicated that the combination of 2095-T6 to 2090-T8 exhibited significantly higher weld shear strength at the interface in comparison to welds of 8090-T6 to 2090-T83. The increased shear strength associated with 2095 is believed to be due to the alloy's ability to precipitation strengthening (naturally age) after welding.

  10. 33. Photographic copy of original construction drawing, dated May 22, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    33. Photographic copy of original construction drawing, dated May 22, 1951 (from paper copy at Engineering Flight, Ellsworth Air Force Base, SD). Readiness hangar structural: details of trusses T2 to T6. - Ellsworth Air Force Base, Readiness Hangar, Kenny Road, southeast corner of interstction with G Avenue, Blackhawk, Meade County, SD

  11. The effects of plasticity in adhesive fracture

    NASA Technical Reports Server (NTRS)

    Chang, M. D.; Devries, K. L.; Williams, M. L.

    1973-01-01

    An energy-balance analysis is presented for adhesive failure in end loaded cantilever beams. The analysis includes the effects of input work, stored strain energy, dissipated plastic energy, and specific adhesive surface energy. Experimental results obtained with 6061-T6 aluminum are presented as evidence for the validity of the approach.-

  12. 46 CFR 160.035-6 - Construction of aluminum oar-, hand-, and motor-propelled lifeboats.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT... gage equals 0.1620. (b) Materials. (1) Plating for shell, air tanks, etc., shall be as shown in Table... sternpost, and the propeller shaft stern tube to the sternpost. When using 6061-T6 aluminum, the welded...

  13. 46 CFR 160.035-6 - Construction of aluminum oar-, hand-, and motor-propelled lifeboats.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT... gage equals 0.1620. (b) Materials. (1) Plating for shell, air tanks, etc., shall be as shown in Table... sternpost, and the propeller shaft stern tube to the sternpost. When using 6061-T6 aluminum, the welded...

  14. 78 FR 68992 - Visas: Documentation of Nonimmigrants-Visa Classification; T Visa Class

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-18

    ... of a T-1 principal alien whom the Secretary of Homeland Security, in consultation with the law... retaliation as a result of the alien's escape from trafficking or cooperation with law enforcement to accompany or follow to join the principal alien. This rule amends 22 CFR 41.12 to include the T-6...

  15. Silencing trehalose-6-phosphate synthase incapacitates adult mosquitoes by interfering with the biosynthetic pathway for flight fuel

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trehalose is a disaccharide comprised of two glucose molecules. It is the main blood sugar of insects and is essential for flight. Trehalose is synthesized by two enzymes: trehalose-6-phosphate synthase (T6PS) converts glucose-6-phosphate to trehalose-6-phosphate, and trehalose-6-phosphate phosphata...

  16. Influence of hot isostatic pressing on the structure and properties of an innovative low-alloy high-strength aluminum cast alloy based on the Al-Zn-Mg-Cu-Ni-Fe system

    NASA Astrophysics Data System (ADS)

    Akopyan, T. K.; Padalko, A. G.; Belov, N. A.

    2015-11-01

    Hot isostatic pressing (HIP) is applied for treatment of castings of innovative low-ally high-strength aluminum alloy, nikalin ATs6N0.5Zh based on the Al-Zn-Mg-Cu-Ni-Fe system. The influence of HIP on the structure and properties of castings is studied by means of three regimes of barometric treatment with different temperatures of isometric holding: t 1 = 505 ± 2°C, p 1 = 100 MPa, τ1 = 3 h (HIP1); t 2 = 525 ± 2°C, p 2 = 100 MPa, τ2 = 3 h (HIP2); and t 3 = 545 ± 2°C, p 3 = 100 MPa, τ3 = 3 h (HIP3). It is established that high-temperature HIP leads to actually complete elimination of porosity and additional improvement of the morphology of second phases. Improved structure after HIP provides improvement properties, especially of plasticity. In particular, after heat treatment according of regime HIP2 + T4 (T4 is natural aging), the alloy plasticity is improved by about two times in comparison with the initial state (from ~6 to 12%). While applying regime HIP3 + T6 (T6 is artificial aging for reaching the maximum strength), the plasticity has improved by more than three times in comparison with the initial state, as after treatment according to regimes HIP1 + T6 and HIP2 + T6 (from ~1.2 to ~5.0%), which are characterized by a lower HIP temperature.

  17. Type VI Secretion System-Associated Gene Clusters Contribute to Pathogenesis of Salmonella enterica Serovar Typhimurium

    PubMed Central

    Mulder, David T.; Cooper, Colin A.

    2012-01-01

    The enteropathogen Salmonella enterica serovar Typhimurium employs a suite of tightly regulated virulence factors within the intracellular compartment of phagocytic host cells resulting in systemic dissemination in mice. A type VI secretion system (T6SS) within Salmonella pathogenicity island 6 (SPI-6) has been implicated in this process; however, the regulatory inputs and the roles of noncore genes in this system are not well understood. Here we describe four clusters of noncore T6SS genes in SPI-6 based on a comparative relationship with the T6SS-3 of Burkholderia mallei and report that the disruption of these genes results in defects in intracellular replication and systemic dissemination in mice. In addition, we show that the expression of the SPI-6-encoded Hcp and VgrG orthologs is enhanced during late stages of macrophage infection. We identify six regions that are transcriptionally active during cell infections and that have regulatory contributions from the regulators of virulence SsrB, PhoP, and SlyA. We show that levels of protein expression are very weak under in vitro conditions and that expression is not enhanced upon the deletion of ssrB, phoP, slyA, qseC, ompR, or hfq, suggesting an unknown activating factor. These data suggest that the SPI-6 T6SS has been integrated into the Salmonella Typhimurium virulence network and customized for host-pathogen interactions through the action of noncore genes. PMID:22493086

  18. Inhibition of Type VI Secretion by an Anti-TssM Llama Nanobody

    PubMed Central

    Spinelli, Silvia; Desmyter, Aline; Le, Thi Thu Hang; Kellenberger, Christine; Douzi, Badreddine; Durand, Eric; Roussel, Alain; Cascales, Eric; Cambillau, Christian

    2015-01-01

    The type VI secretion system (T6SS) is a secretion pathway widespread in Gram-negative bacteria that targets toxins in both prokaryotic and eukaryotic cells. Although most T6SSs identified so far are involved in inter-bacterial competition, a few are directly required for full virulence of pathogens. The T6SS comprises 13 core proteins that assemble a large complex structurally and functionally similar to a phage contractile tail structure anchored to the cell envelope by a trans-membrane spanning stator. The central part of this stator, TssM, is a 1129-amino-acid protein anchored in the inner membrane that binds to the TssJ outer membrane lipoprotein. In this study, we have raised camelid antibodies against the purified TssM periplasmic domain. We report the crystal structure of two specific nanobodies that bind to TssM in the nanomolar range. Interestingly, the most potent nanobody, nb25, competes with the TssJ lipoprotein for TssM binding in vitro suggesting that TssJ and the nb25 CDR3 loop share the same TssM binding site or causes a steric hindrance preventing TssM-TssJ complex formation. Indeed, periplasmic production of the nanobodies displacing the TssM-TssJ interaction inhibits the T6SS function in vivo. This study illustrates the power of nanobodies to specifically target and inhibit bacterial secretion systems. PMID:25811612

  19. Localized pruritus-notalgia paresthetica.

    PubMed

    Massey, E W; Pleet, A B

    1979-08-01

    Notalgia paresthetica, possibly an isolated sensory neuropathy involving the posterior primary rami of thoracic nerves T2 through T6, and appearing as pruritus of the back, is apt to be encountered by both dermatologists and neurologists. Two cases illustrate this disorder. PMID:464629

  20. Notalgia paresthetica owing to compression neuropathy: case presentation including electrodiagnostic studies.

    PubMed

    Streib, E W; Sun, S F

    1981-01-01

    A patient developed numbness over the upper part of his back, following prolonged bed rest in a supine position. Sweat test, clinical and electromyographic examination indicated neuropathies of the primary dorsal rami of the spinal nerves from T2 to T6. The literature is reviewed and anatomical considerations discussed. PMID:7202444

  1. 27 CFR 9.120 - Sierra Foothills.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... the county line meets Dry Creek in the northwestern corner of T. 5 N., R. 9 E.; (9) Then in a south... County line meets the western border of the boundary of the Sierra National Forest in T. 6S, R. 20 E... borders of the Sierra and Stanislaus National Forests in Mariposa County (see “San Jose” map); (16)...

  2. 27 CFR 9.120 - Sierra Foothills.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... the county line meets Dry Creek in the northwestern corner of T. 5 N., R. 9 E.; (9) Then in a south... County line meets the western border of the boundary of the Sierra National Forest in T. 6S, R. 20 E... borders of the Sierra and Stanislaus National Forests in Mariposa County (see “San Jose” map); (16)...

  3. The Fungus Trichoderma Regulates Submerged Conidiation Using the Steroid Pregnenolone.

    PubMed

    Lin, Hui; Travisano, Michael; Kazlauskas, Romas J

    2016-09-16

    In previous work, we evolved a population of Trichoderma citrinoviride in liquid cultures to speed up its asexual development cycle. The evolved population, called T-6, formed conidia 3 times sooner and in >1000-fold greater numbers. Here, we identify the steroid pregnenolone as a molecular signal for this different behavior. Media in which the ancestral T. citrinoviride population was grown (called ancestral spent media) contained a submerged conidiation inhibitor. Growing the evolved population T-6 in ancestral spent media eliminated the abundant formation of conidia. This inhibition depended on the amount and age of the ancestral spent medium and the time that the ancestral spent medium was added to the T-6 culture. Fractionation of the ancestral spent medium identified a hydrophobic inhibiting compound with a molecular weight less than 2000 g/mol. A combination of GC-MS, ELISA, and reaction with cholesterol oxidase identified it as pregnenolone. The addition of pregnenolone to cultures of T-6 inhibited submerged conidiation by inhibiting formation of conidiophores, while 10 other analogous steroids did not. Pregnenolone also inhibited submerged conidiation of Fusarium graminearum PH-1, a plant pathogen that causes head blight in wheat and barley. This identification of steroids as signal molecules in fungi creates opportunities to disrupt this signaling to control fungal behavior. PMID:27413801

  4. Transcriptional frameshifting rescues Citrobacter rodentium type VI secretion by the production of two length variants from the prematurely interrupted tssM gene.

    PubMed

    Gueguen, Erwan; Wills, Norma M; Atkins, John F; Cascales, Eric

    2014-12-01

    The Type VI secretion system (T6SS) mediates toxin delivery into both eukaryotic and prokaryotic cells. It is composed of a cytoplasmic structure resembling the tail of contractile bacteriophages anchored to the cell envelope through a membrane complex composed of the TssL and TssM inner membrane proteins and of the TssJ outer membrane lipoprotein. The C-terminal domain of TssM is required for its interaction with TssJ, and for the function of the T6SS. In Citrobacter rodentium, the tssM1 gene does not encode the C-terminal domain. However, the stop codon is preceded by a run of 11 consecutive adenosines. In this study, we demonstrate that this poly-A tract is a transcriptional slippery site that induces the incorporation of additional adenosines, leading to frameshifting, and hence the production of two TssM1 variants, including a full-length canonical protein. We show that both forms of TssM1, and the ratio between these two forms, are required for the function of the T6SS in C. rodentium. Finally, we demonstrate that the tssM gene associated with the Yersinia pseudotuberculosis T6SS-3 gene cluster is also subjected to transcriptional frameshifting.

  5. Regulation of Type VI Secretion Gene Clusters by σ54 and Cognate Enhancer Binding Proteins▿†

    PubMed Central

    Bernard, Christophe S.; Brunet, Yannick R.; Gavioli, Marthe; Lloubès, Roland; Cascales, Eric

    2011-01-01

    Type VI secretion systems (T6SS) are bacteriophage-derived macromolecular machines responsible for the release of at least two proteins in the milieu, which are thought to form an extracellular appendage. Although several T6SS have been shown to be involved in the virulence of animal and plant pathogens, clusters encoding these machines are found in the genomes of most species of Gram-negative bacteria, including soil, marine, and environmental isolates. T6SS have been associated with several phenotypes, ranging from virulence to biofilm formation or stress sensing. Their various environmental niches and large diversity of functions are correlated with their broad variety of regulatory mechanisms. Using a bioinformatic approach, we identified several clusters, including those of Vibrio cholerae, Aeromonas hydrophila, Pectobacterium atrosepticum, Pseudomonas aeruginosa, Pseudomonas syringae pv. tomato, and a Marinomonas sp., which possess typical −24/−12 sequences, recognized by the alternate sigma factor sigma 54 (σ54 or σN). σ54, which directs the RNA polymerase to these promoters, requires the action of a bacterial enhancer binding protein (bEBP), which binds to cis-acting upstream activating sequences. Putative bEBPs are encoded within the T6SS gene clusters possessing σ54 boxes. Using in vitro binding experiments and in vivo reporter fusion assays, we showed that the expression of these clusters is dependent on both σ54 and bEBPs. PMID:21378190

  6. Type Six Secretion System of Bordetella bronchiseptica and Adaptive Immune Components Limit Intracellular Survival During Infection.

    PubMed

    Bendor, Liron; Weyrich, Laura S; Linz, Bodo; Rolin, Olivier Y; Taylor, Dawn L; Goodfield, Laura L; Smallridge, William E; Kennett, Mary J; Harvill, Eric T

    2015-01-01

    The Type Six Secretion System (T6SS) is required for Bordetella bronchiseptica cytotoxicity, cytokine modulation, infection, and persistence. However, one-third of recently sequenced Bordetella bronchiseptica strains of the predominantly human-associated Complex IV have lost their T6SS through gene deletion or degradation. Since most human B. bronchiseptica infections occur in immunocompromised patients, we determine here whether loss of Type Six Secretion is beneficial to B. bronchiseptica during infection of immunocompromised mice. Infection of mice lacking adaptive immunity (Rag1-/- mice) with a T6SS-deficient mutant results in a hypervirulent phenotype that is characterized by high numbers of intracellular bacteria in systemic organs. In contrast, wild-type B. bronchiseptica kill their eukaryotic cellular hosts via a T6SS-dependent mechanism that prevents survival in systemic organs. High numbers of intracellular bacteria recovered from immunodeficient mice but only low numbers from wild-type mice demonstrates that B. bronchiseptica survival in an intracellular niche is limited by B and T cell responses. Understanding the nature of intracellular survival during infection, and its effects on the generation and function of the host immune response, are important to contain and control the spread of Bordetella-caused disease.

  7. Depressive Symptoms and Associated Psychosocial Factors among Adolescent Survivors 30 Months after 2008 Wenchuan Earthquake: A Follow-Up Study

    PubMed Central

    Shi, Xuliang; Yu, Nancy X.; Zhou, Ya; Geng, Fulei; Fan, Fang

    2016-01-01

    Purpose: This study longitudinally investigated the changes of depressive symptoms among adolescent survivors over 2 years and a half after the 2008 Wenchuan earthquake in China, as well as the predictive effects of demographic characteristics, earthquake exposure, negative life events, social support, and dispositional resilience on the risk of depressive symptoms at two time points after the earthquake. Methods: Participants were 1573 adolescent survivors (720 males and 853 females, mean age at initial survey = 15.00 ± 1.26 years), whose depressive symptoms were assessed at 6 months (T6m) and 30 months (T30m) post-earthquake. Data on demographics, earthquake exposure, and dispositional resilience were collected at T6m. Negative life events and social support were measured at T6m and 24 months (T24m) post-earthquake. Results: The prevalence rates of probable depression, 27.5 at T6m and 27.2% at T30m, maintained relatively stable over time. Female gender was related with higher risk of depressive symptoms at both T6m and T30m, while being only child could only predict higher risk of depressive symptoms at T30m. Negative life events and social support at T6m, as well as earthquake exposure, were concurrently associated with increased risk of depressive symptoms at T6m, but not associated with the risk of depressive symptoms at T30m, while negative life events and social support at T24m could predict depressive symptoms at T30m, all of which suggested that these variables may have strong but short-term effect on adolescents’ depressive symptoms post-earthquake. Besides, dispositional resilience was evidenced as a relatively stable negative predictor for depressive symptoms. Conclusions: These findings could inform mental health professionals regarding how to screen adolescent survivors at high risk for depression, so as to provide them with timely and appropriate mental health services based on the identified risk and protective factors for depressive symptoms

  8. Effects of black pepper (piper nigrum), turmeric powder (curcuma longa) and coriander seeds (coriandrum sativum) and their combinations as feed additives on growth performance, carcass traits, some blood parameters and humoral immune response of broiler chickens.

    PubMed

    Abou-Elkhair, R; Ahmed, H A; Selim, S

    2014-06-01

    Different herbs and spices have been used as feed additives for various purposes in poultry production. This study was conducted to assess the effect of feed supplemented with black pepper (Piper nigrum), turmeric powder (Curcuma longa), coriander seeds (Coriandrum sativum) and their combinations on the performance of broilers. A total of 210 (Cobb) one-d-old chicks were divided into seven groups of 30 birds each. The treatments were: a control group received no supplement, 0.5% black pepper (T1), 0.5% turmeric powder (T2), 2% coriander seeds (T3), a mixture of 0.5% black pepper and 0.5% turmeric powder (T4), a mixture of 0.5% black pepper and 2% coriander seed (T5), and a mixture of 0.5% black pepper, 0.5% turmeric powder and 2% coriander seeds (T6). Higher significant values of body weight gain during the whole period of 5 weeks (p<0.001) were observed in broilers on T1, T3, T5, and T6 compared to control. Dietary supplements with T1, T2, T3, and T6 improved the cumulative G:F of broilers during the whole period of 5 weeks (p<0.001) compared with control. The dressing percentage and edible giblets were not influenced by dietary supplements, while higher values of relative weight of the liver (p<0.05) were obtained in T5 and T6 compared to control. The addition of feed supplements in T5 and T6 significantly increased serum total protein and decreased serum glucose, triglycerides and alkaline phosphatase concentrations compared with the control group (p<0.05). Broilers on T6 showed significant decrease in the serum glutamate pyruvate transaminase concentration (p<0.05) compared to control. The broilers having T5 and T6 supplemented feed had relatively greater antibody titre (p<0.001) at 35 d of age than control. It is concluded that dietary supplements with black pepper or coriander seeds or their combinations enhanced the performance and health status of broiler chickens.

  9. Changes in blood testosterone concentrations after surgical and chemical sterilization of male free-roaming dogs in southern Chile.

    PubMed

    Vanderstichel, R; Forzán, M J; Pérez, G E; Serpell, J A; Garde, E

    2015-04-01

    There is a growing interest in chemical sterilization as an alternative to surgical castration in large-scale sterilization campaigns to control canine populations. An important step toward understanding the short-term and long-term effects of chemical sterilants is to determine their impact on blood testosterone concentrations, particularly as these could influence dog behavior after treatment. A field trial was conducted with 118 free-roaming male dogs in the Chilean Patagonia, where 36 dogs were chemically sterilized using EsterilSol, 39 dogs were surgically castrated, and 43 dogs remained intact as controls. Blood testosterone levels were determined at four time periods: on enrollment 6 months before treatment (t-6m), at the time of treatment (t0, within one hour after surgical castration or chemical sterilization and during a concurrent 2-week period for the control group), four (t+4m), and six (t+6m) months after treatment. Intrinsic and temporal factors were evaluated; age was significantly associated with testosterone, where dogs 2- to 4-year-old had the highest testosterone concentrations (P = 0.036), whereas body weight and body condition scores were not associated with testosterone; testosterone concentration was not influenced by time of day, month, or season. After treatment (t+4m and t+6m), all of the surgically castrated dogs had testosterone concentrations below 1.0 ng/mL. On the basis of this cut point (<1 ng/mL), testosterone remained unchanged in 66% of the chemically sterilized dogs at both t+4m and t+6m; it remained low for 22% of dogs at both t+4m and t+6m; it was unchanged at t+4m but low at t+6m in 9% of dogs; and, it was low at t+4m but reverted back to unchanged at t+6m in one dog (3%). Incidentally, testosterone in chemically sterilized dogs increased dramatically within 1 hour of treatment (t0), more than doubling (131%) the concentration of control dogs at the time of treatment (t0), likely because of severe necrosis of

  10. Effects of Black Pepper (Piper Nigrum), Turmeric Powder (Curcuma Longa) and Coriander Seeds (Coriandrum Sativum) and Their Combinations as Feed Additives on Growth Performance, Carcass Traits, Some Blood Parameters and Humoral Immune Response of Broiler Chickens

    PubMed Central

    Abou-Elkhair, R.; Ahmed, H. A.; Selim, S.

    2014-01-01

    Different herbs and spices have been used as feed additives for various purposes in poultry production. This study was conducted to assess the effect of feed supplemented with black pepper (Piper nigrum), turmeric powder (Curcuma longa), coriander seeds (Coriandrum sativum) and their combinations on the performance of broilers. A total of 210 (Cobb) one-d-old chicks were divided into seven groups of 30 birds each. The treatments were: a control group received no supplement, 0.5% black pepper (T1), 0.5% turmeric powder (T2), 2% coriander seeds (T3), a mixture of 0.5% black pepper and 0.5% turmeric powder (T4), a mixture of 0.5% black pepper and 2% coriander seed (T5), and a mixture of 0.5% black pepper, 0.5% turmeric powder and 2% coriander seeds (T6). Higher significant values of body weight gain during the whole period of 5 weeks (p<0.001) were observed in broilers on T1, T3, T5, and T6 compared to control. Dietary supplements with T1, T2, T3, and T6 improved the cumulative G:F of broilers during the whole period of 5 weeks (p<0.001) compared with control. The dressing percentage and edible giblets were not influenced by dietary supplements, while higher values of relative weight of the liver (p<0.05) were obtained in T5 and T6 compared to control. The addition of feed supplements in T5 and T6 significantly increased serum total protein and decreased serum glucose, triglycerides and alkaline phosphatase concentrations compared with the control group (p<0.05). Broilers on T6 showed significant decrease in the serum glutamate pyruvate transaminase concentration (p<0.05) compared to control. The broilers having T5 and T6 supplemented feed had relatively greater antibody titre (p<0.001) at 35 d of age than control. It is concluded that dietary supplements with black pepper or coriander seeds or their combinations enhanced the performance and health status of broiler chickens. PMID:25050023

  11. Fine tuning of trehalose biosynthesis and hydrolysis as novel tools for the generation of abiotic stress tolerant plants

    PubMed Central

    Delorge, Ines; Janiak, Michal; Carpentier, Sebastien; Van Dijck, Patrick

    2014-01-01

    The impact of abiotic stress on plant growth and development has been and still is a major research topic. An important pathway that has been linked to abiotic stress tolerance is the trehalose biosynthetic pathway. Recent findings showed that trehalose metabolism is also important for normal plant growth and development. The intermediate compound – trehalose-6-phosphate (T6P) – is now confirmed to act as a sensor for available sucrose, hereby directly influencing the type of response to the changing environmental conditions. This is possible because T6P and/or trehalose or their biosynthetic enzymes are part of complex interaction networks with other crucial hormone and sugar-induced signaling pathways, which may function at different developmental stages. Because of its effect on plant growth and development, modification of trehalose biosynthesis, either at the level of T6P synthesis, T6P hydrolysis, or trehalose hydrolysis, has been utilized to try to improve crop yield and biomass. It was shown that alteration of the amounts of either T6P and/or trehalose did result in increased stress tolerance, but also resulted in many unexpected phenotypic alterations. A main challenge is to characterize the part of the signaling pathway resulting in improved stress tolerance, without affecting the pathways resulting in the unwanted phenotypes. One such specific pathway where modification of trehalose metabolism improved stress tolerance, without any side effects, was recently obtained by overexpression of trehalase, which results in a more sensitive reaction of the stomatal guard cells and closing of the stomata under drought stress conditions. We have used the data that have been obtained from different studies to generate the optimal plant that can be constructed based on modifications of trehalose metabolism. PMID:24782885

  12. Impact Welding of Aluminum to Copper and Stainless Steel by Vaporizing Foil Actuator: Effect of Heat Treatment Cycles on Mechanical Properties and Microstructure

    NASA Astrophysics Data System (ADS)

    Vivek, Anupam; Hansen, Steven; Benzing, Jake; He, Mei; Daehn, Glenn

    2015-10-01

    This work studies the mechanical property effect of microstructure on impact welds of aluminum alloy AA6061 with both copper alloy Cu 110 and stainless steel AISI 304. AA6061-T6 and T4 temper aluminum sheets of 1 mm thickness were launched toward copper and stainless steel targets using the vaporizing foil actuator technique. Flyer plate velocities, measured via photonic Doppler velocimetry, were observed to be approximately 800 m/s. The welded aluminum-copper samples were subjected to instrumented peel testing, microhardness testing, energy-dispersive X-ray spectroscopy, and scanning electron microscopy. The welded joints exhibited cracks through their continuous intermetallic layers. The cracks were impeded upon encountering a ductile metallic wave. The welds created with T6 temper flyer sheets were found to have smaller intermetallic-free and wavy interface regions as compared to those created with T4 temper flyer sheets. Peel strength tests of the two weld combinations resulted in failure along the interface in the case of the T6 flyer welds, while the failure generally occurred in the parent aluminum for the T4 temper flyer welds. Half of the T4 flyer welds were subjected to aging for 18 h at 433 K (160 °C) to convert the aluminum sheet to the T6 condition. Although the aged flyer material did not attain the hardness of the as-received T6 material, it was found to be significantly stronger than the T4 material. These welds retained their strength after the aging process, and diffusion across the interface was minimal. The welded aluminum-stainless steel samples were analyzed on a more basic level than aluminum-copper samples, but were found to exhibit similar results.

  13. A longitudinal study of streptococcus pneumoniae carriage in healthy children in the 13-valent pneumococcal conjugate vaccine era

    PubMed Central

    Mameli, Chiara; Fabiano, Valentina; Daprai, Laura; Bedogni, Giorgio; Faccini, Marino; Garlaschi, Maria Laura; Penagini, Francesca; Dilillo, Dario; Torresani, Erminio; Gramegna, Maria; Zuccotti, Gian Vincenzo

    2015-01-01

    Few epidemiological data are available after the introduction of the 13-valent pneumococcal vaccine (PCV13) in 2010. We performed repeat nasopharyngeal swabs and evaluated the serotype distribution of Streptococcus pneumoniae (SP) and its association with PCV13 vaccine status in healthy Italian children aged 3–59 months. SP serotypes were assessed by the Quellung reaction. 618 children appropriately (28%) or incompletely (72%) vaccinated for age with PCV13 were available at baseline (T0). 515 were re-evaluated at 6 months from baseline (T6) and 436 at 12 months from baseline (T12). The percentage of appropriately vaccinated subjects at T0, T6 and T12 was 28%, 67% and 92%, respectively. Random effects logistic regression models with robust 95% confidence intervals was used to estimate the time-related changes in SP and PCV13 carriage and marginal probabilities were obtained from such models. The age-corrected probability of SP carriage was 0.31 (95% CI 0.22 - 0.41) at T0, 0.32 (0.24 - 0.40) at T6 and 0.28 (0.20 - 0.35) at T12. The probability of PCV13 serotypes carriage was 0.025 (0.001 - 0.050) at T0, 0.018 (0.001 - 0.039) at T6 and 0.010 (0.001 - 0.023) at T12. A decrease in PCV13 serotypes and a shift in non-PCV13 serotypes colonization was observed. In particular, the 15A serotype accounted for 4%, 8% and 23% of SP isolates at T0, T6 and T12, respectively. In conclusion, the benefits of the PCV13 vaccination on SP carriage increase with increasing coverage rates. The shift of SP isolates toward non-PCV13 serotypes needs to be studied further. PMID:25751237

  14. Coevolution of the ATPase ClpV, the sheath proteins TssB and TssC, and the accessory protein TagJ/HsiE1 distinguishes type VI secretion classes.

    PubMed

    Förster, Andreas; Planamente, Sara; Manoli, Eleni; Lossi, Nadine S; Freemont, Paul S; Filloux, Alain

    2014-11-21

    The type VI secretion system (T6SS) is a bacterial nanomachine for the transport of effector molecules into prokaryotic and eukaryotic cells. It involves the assembly of a tubular structure composed of TssB and TssC that is similar to the tail sheath of bacteriophages. The sheath contracts to provide the energy needed for effector delivery. The AAA(+) ATPase ClpV disassembles the contracted sheath, which resets the systems for reassembly of an extended sheath that is ready to fire again. This mechanism is crucial for T6SS function. In Vibrio cholerae, ClpV binds the N terminus of TssC within a hydrophobic groove. In this study, we resolved the crystal structure of the N-terminal domain of Pseudomonas aeruginosa ClpV1 and observed structural alterations in the hydrophobic groove. The modification in the ClpV1 groove is matched by a change in the N terminus of TssC, suggesting the existence of distinct T6SS classes. An accessory T6SS component, TagJ/HsiE, exists predominantly in one of the classes. Using bacterial two-hybrid approaches, we showed that the P. aeruginosa homolog HsiE1 interacts strongly with ClpV1. We then resolved the crystal structure of HsiE1 in complex with the N terminus of HsiB1, a TssB homolog and component of the contractile sheath. Phylogenetic analysis confirmed that these differences distinguish T6SS classes that resulted from a functional co-evolution between TssB, TssC, TagJ/HsiE, and ClpV. The interaction of TagJ/HsiE with the sheath as well as with ClpV suggests an alternative mode of disassembly in which HsiE recruits the ATPase to the sheath. PMID:25305017

  15. Genomic and Functional Analysis of the Type VI Secretion System in Acinetobacter

    PubMed Central

    Weber, Brent S.; Miyata, Sarah T.; Iwashkiw, Jeremy A.; Mortensen, Brittany L.; Skaar, Eric P.; Pukatzki, Stefan; Feldman, Mario F.

    2013-01-01

    The genus Acinetobacter is comprised of a diverse group of species, several of which have raised interest due to potential applications in bioremediation and agricultural purposes. In this work, we show that many species within the genus Acinetobacter possess the genetic requirements to assemble a functional type VI secretion system (T6SS). This secretion system is widespread among Gram negative bacteria, and can be used for toxicity against other bacteria and eukaryotic cells. The most studied species within this genus is A. baumannii, an emerging nosocomial pathogen that has become a significant threat to healthcare systems worldwide. The ability of A. baumannii to develop multidrug resistance has severely reduced treatment options, and strains resistant to most clinically useful antibiotics are frequently being isolated. Despite the widespread dissemination of A. baumannii, little is known about the virulence factors this bacterium utilizes to cause infection. We determined that the T6SS is conserved and syntenic among A. baumannii strains, although expression and secretion of the hallmark protein Hcp varies between strains, and is dependent on TssM, a known structural protein required for T6SS function. Unlike other bacteria, A. baumannii ATCC 17978 does not appear to use its T6SS to kill Escherichia coli or other Acinetobacter species. Deletion of tssM does not affect virulence in several infection models, including mice, and did not alter biofilm formation. These results suggest that the T6SS fulfils an important but as-yet-unidentified role in the various lifestyles of the Acinetobacter spp. PMID:23365692

  16. Sex-specific antennal sensory system in the ant Camponotus japonicus: glomerular organizations of antennal lobes.

    PubMed

    Nakanishi, Aki; Nishino, Hiroshi; Watanabe, Hidehiro; Yokohari, Fumio; Nishikawa, Michiko

    2010-06-15

    Ants have well-developed chemosensory systems for social lives. The goal of our study is to understand the functional organization of the ant chemosensory system based on caste- and sex-specific differences. Here we describe the common and sex-specific glomerular organizations in the primary olfactory center, the antennal lobe of the carpenter ant Camponotus japonicus. Differential labeling of the two antennal nerves revealed distinct glomerular clusters innervated by seven sensory tracts (T1-T7 from ventral to dorsal) in the antennal lobe. T7 innervated 10 glomeruli, nine of which received thick axon terminals almost exclusively from the ventral antennal nerve. Coelocapitular (hygro-/thermoreceptive), coeloconic (thermoreceptive), and ampullaceal (CO2-receptive) sensilla, closely appositioned in the flagellum, housed one or three large sensory neurons supplying thick axons exclusively to the ventral antennal nerve. These axons, therefore, were thought to project into T7 glomeruli in all three castes. Workers and virgin females had about 140 T6 glomeruli, whereas males completely lacked these glomeruli. Female-specific basiconic sensilla (cuticular hydrocarbon-receptive) contained over 130 sensory neurons and were completely lacking in males' antennae. These sensory neurons may project into T6 glomeruli in the antennal lobe of workers and virgin females. Serotonin-immunopositive neurons innervated T1-T5 and T7 glomeruli but not T6 glomeruli in workers and virgin females. Because males had no equivalents to T6 glomeruli, serotonin-immunopositive neurons appeared to innervate all glomeruli in the male's antennal lobe. T6 glomeruli in workers and virgin females are therefore female-specific and may have functions related to female-specific tasks in the colony rather than sexual behaviors.

  17. Sex-specific antennal sensory system in the ant Camponotus japonicus: glomerular organizations of antennal lobes.

    PubMed

    Nakanishi, Aki; Nishino, Hiroshi; Watanabe, Hidehiro; Yokohari, Fumio; Nishikawa, Michiko

    2010-06-15

    Ants have well-developed chemosensory systems for social lives. The goal of our study is to understand the functional organization of the ant chemosensory system based on caste- and sex-specific differences. Here we describe the common and sex-specific glomerular organizations in the primary olfactory center, the antennal lobe of the carpenter ant Camponotus japonicus. Differential labeling of the two antennal nerves revealed distinct glomerular clusters innervated by seven sensory tracts (T1-T7 from ventral to dorsal) in the antennal lobe. T7 innervated 10 glomeruli, nine of which received thick axon terminals almost exclusively from the ventral antennal nerve. Coelocapitular (hygro-/thermoreceptive), coeloconic (thermoreceptive), and ampullaceal (CO2-receptive) sensilla, closely appositioned in the flagellum, housed one or three large sensory neurons supplying thick axons exclusively to the ventral antennal nerve. These axons, therefore, were thought to project into T7 glomeruli in all three castes. Workers and virgin females had about 140 T6 glomeruli, whereas males completely lacked these glomeruli. Female-specific basiconic sensilla (cuticular hydrocarbon-receptive) contained over 130 sensory neurons and were completely lacking in males' antennae. These sensory neurons may project into T6 glomeruli in the antennal lobe of workers and virgin females. Serotonin-immunopositive neurons innervated T1-T5 and T7 glomeruli but not T6 glomeruli in workers and virgin females. Because males had no equivalents to T6 glomeruli, serotonin-immunopositive neurons appeared to innervate all glomeruli in the male's antennal lobe. T6 glomeruli in workers and virgin females are therefore female-specific and may have functions related to female-specific tasks in the colony rather than sexual behaviors. PMID:20437523

  18. T346Hunter: a novel web-based tool for the prediction of type III, type IV and type VI secretion systems in bacterial genomes.

    PubMed

    Martínez-García, Pedro Manuel; Ramos, Cayo; Rodríguez-Palenzuela, Pablo

    2015-01-01

    T346Hunter (Type Three, Four and Six secretion system Hunter) is a web-based tool for the identification and localisation of type III, type IV and type VI secretion systems (T3SS, T4SS and T6SS, respectively) clusters in bacterial genomes. Non-flagellar T3SS (NF-T3SS) and T6SS are complex molecular machines that deliver effector proteins from bacterial cells into the environment or into other eukaryotic or prokaryotic cells, with significant implications for pathogenesis of the strains encoding them. Meanwhile, T4SS is a more functionally diverse system, which is involved in not only effector translocation but also conjugation and DNA uptake/release. Development of control strategies against bacterial-mediated diseases requires genomic identification of the virulence arsenal of pathogenic bacteria, with T3SS, T4SS and T6SS being major determinants in this regard. Therefore, computational methods for systematic identification of these specialised machines are of particular interest. With the aim of facilitating this task, T346Hunter provides a user-friendly web-based tool for the prediction of T3SS, T4SS and T6SS clusters in newly sequenced bacterial genomes. After inspection of the available scientific literature, we constructed a database of hidden Markov model (HMM) protein profiles and sequences representing the various components of T3SS, T4SS and T6SS. T346Hunter performs searches of such a database against user-supplied bacterial sequences and localises enriched regions in any of these three types of secretion systems. Moreover, through the T346Hunter server, users can visualise the predicted clusters obtained for approximately 1700 bacterial chromosomes and plasmids. T346Hunter offers great help to researchers in advancing their understanding of the biological mechanisms in which these sophisticated molecular machines are involved. T346Hunter is freely available at http://bacterial-virulence-factors.cbgp.upm.es/T346Hunter.

  19. The trehalose pathway in maize: conservation and gene regulation in response to the diurnal cycle and extended darkness

    PubMed Central

    Henry, Clémence; Bledsoe, Samuel W.; Siekman, Allison; Kollman, Alec; Waters, Brian M.; Feil, Regina; Stitt, Mark; Lagrimini, L. Mark

    2014-01-01

    Energy resources in plants are managed in continuously changing environments, such as changes occurring during the day/night cycle. Shading is an environmental disruption that decreases photosynthesis, compromises energy status, and impacts on crop productivity. The trehalose pathway plays a central but not well-defined role in maintaining energy balance. Here, we characterized the maize trehalose pathway genes and deciphered the impacts of the diurnal cycle and disruption of the day/night cycle on trehalose pathway gene expression and sugar metabolism. The maize genome encodes 14 trehalose-6-phosphate synthase (TPS) genes, 11 trehalose-6-phosphate phosphatase (TPP) genes, and one trehalase gene. Transcript abundance of most of these genes was impacted by the day/night cycle and extended dark stress, as were sucrose, hexose sugars, starch, and trehalose-6-phosphate (T6P) levels. After extended darkness, T6P levels inversely followed class II TPS and sucrose non-fermenting-related protein kinase 1 (SnRK1) target gene expression. Most significantly, T6P no longer tracked sucrose levels after extended darkness. These results showed: (i) conservation of the trehalose pathway in maize; (ii) that sucrose, hexose, starch, T6P, and TPS/TPP transcripts respond to the diurnal cycle; and(iii) that extended darkness disrupts the correlation between T6P and sucrose/hexose pools and affects SnRK1 target gene expression. A model for the role of the trehalose pathway in sensing of sucrose and energy status in maize seedlings is proposed. PMID:25271261

  20. Factors that influence healing of chronic venous leg ulcers: a retrospective cohort*

    PubMed Central

    Scotton, Marilia Formentini; Miot, Hélio Amante; Abbade, Luciana Patricia Fernandes

    2014-01-01

    BACKGROUND Venous ulcers have a significant impact on patient quality of life, and constitute a worldwide public health problem. Treatment is complex, with high failure rates. OBJECTIVES To identify clinical and therapeutic factors that influence healing of venous ulcers. METHODS Retrospective cohort study of patients with venous ulcers. Ulcer area was measured at the first visit (T0) and after 6 months (T6) and 1 year (T12). A reduction in ulcer area of 50% or more at T6 and T12 was the outcome of interest, weighted by clinical, demographic and treatment aspects. RESULTS Ninety-four patients were included (137 ulcers). A reduction in ulcer area of 50% or more was seen in 40.1% of patients (95%CI 31.9 to 48.4%) at T6 and 49.6% (95%CI 41.2 to 58.1%) at T12. Complete healing occurred in 16.8% (95%CI 10.5 to 23.1%) at T6 and 27% (95%CI 19.5 to 39.5%) at T12. The lowest ulcer area reductions at T6 were associated with longstanding ulcer (RR=0.95; 95%CI 0.91 to 0.98), poor adherence to compression therapy (RR=4.04; 95%CI 1.31 to 12.41), and infection episodes (RR= 0.42; 95%CI 0.23 to 0.76). At T12, lower reductions were associated with longstanding ulcer (RR=0.95; 95%CI 0.92 to 0.98), longer topical antibiotic use (RR=0.93; 95%CI 0.87 to 0.99), and systemic antibiotic use (RR=0.63; 95%CI 0.40 to 0.99). CONCLUSIONS Longstanding ulcer, infection, poor adherence to compression therapy, and longer topical and systemic antibiotic use were independently correlated with worse healing rates. PMID:24937814

  1. Trehalose-6-phosphate and SNF1-related protein kinase 1 are involved in the first-fruit inhibition of cucumber.

    PubMed

    Zhang, ZhiPing; Deng, Yukun; Song, Xingxing; Miao, Minmin

    2015-04-01

    In cucumber (Cucumis sativus L.), the preexisting fruits inhibit the growth of subsequent fruits. To study the mechanism underlying this phenomenon, we examined the sink activity, the level of free sugars, and the activity of SNF1-related protein kinase 1 (SnRK1) in the peduncles of two types of fruits. In the two-fruit cucumber plants, the growth rate and sink activity [evaluated by alkaline alpha-galactosidase (CsAGA) activity in the peduncle] of the first fruit were greater than those of the second fruit. The (14)C-labeling experiment revealed that assimilates produced by the leaves closer to the second fruit tended to move to the first fruit. Sucrose and trehalose-6-phosphate (T6P) levels in the peduncle of the first fruit were higher than those in the peduncle of the second fruit. The SnRK1 activity was lower in the peduncle of the first fruit than in that of the second fruit at 0-8 days after anthesis. The growth rate and sink activity of the second fruit were enhanced after the removal of the first fruit or after treatment with 6-benzyl aminopurine, as determined by comparison with an increase in the sucrose and T6P levels and a decrease in the SnRK1 activity in its peduncle. The SnRK1 activity was inhibited by T6P in an in vitro kinase assay, and the mRNA level of CsAGA1 in cucumber calli was up-regulated by exogenous trehalose treatment, confirming that the SnRK1 activity and CsAGA1 expression can be regulated by T6P levels. Our results suggest that the T6P- and SnRK1-mediated signaling functions are involved in the regulation of first-fruit inhibition in cucumber plants.

  2. New approach for the strategic control of gastrointestinal nematodes in grazed beef cattle during the growing phase in central Brazil.

    PubMed

    Heckler, R P; Borges, D G L; Vieira, M C; Conde, M H; Green, M; Amorim, M L; Echeverria, J T; Oliveira, T L; Moro, E; Van Onselen, V J; Borges, F A

    2016-05-15

    We evaluated the effect of different treatment protocols against gastrointestinal nematodes in Nelore beef cattle during the growing phase in the municipality of Terenos, MS, in central Brazil from May 2013 to April 2014 and from May 2014 to April 2015. Ninety-six Nelore calves were kept on Brachiaria brizantha grass during each trial period and were distributed into six experimental groups (replicate paddocks for each group) based on live weight and the number of eggs per gram of feces (EPG): T1 (control)-treated in May, July and September with a saline solution; T2-treated in May and November with 700 μg/kg doramectin; T3-treated in May (doramectin), July (4.7 mg/kg levamisole phosphate) and September (doramectin); T4-treated in May (doramectin), July (200 μg/kg moxidectin) and September (doramectin); T5-treated in May (doramectin), August (levamisole phosphate) and November (doramectin) and T6-treated in May (doramectin), August (moxidectin) and November (doramectin). The calves were weighed and feces were collected (for faecal culture and EPG counts) from calves every 28 days, concomitantly with the collection of forage samples. The efficacies of doramectin, moxidectin and levamisole were low, at 69.2, 65.9 and 69.4% in the first and 13.8, 92.6, and 76.5% in the second experimental periods, respectively, but only the untreated animals lost weight during the dry season. Final weight gains did not differ significantly (p>0.05) among the animals in T2 (120.8 kg), T3 (131.4 kg), T4 (131.2 kg) and T5 (134.4 kg). T6 was the only group with a significantly higher final weight gain (140.9 kg) compared to the protocol with two annual dosages (T2). The weight gain was 31.9% higher in T6 than in the untreated animals (T1). None of the protocols affected the number of larvae on the pasture. Body weight was significantly and negatively (r=-0.65) correlated with EPG counts, which were significantly lower in June (T2, T3, T4 and T6), August (T3), September (T5 and T6

  3. Preliminary Investigation of Zircaloy-4 as a Research Reactor Cladding Material

    SciTech Connect

    Brian K Castle

    2012-05-01

    As part of a scoping study for the ATR fuel conversion project, an initial comparison of the material properties of Zircaloy-4 and Aluminum-6061 (T6 and O-temper) is performed to provide a preliminary evaluation of Zircaloy-4 for possible inclusion as a candidate cladding material for ATR fuel elements. The current fuel design for the ATR uses Aluminum 6061 (T6 and O temper) as a cladding and structural material in the fuel element and to date, no fuel failures have been reported. Based on this successful and longstanding operating history, Zircaloy-4 properties will be evaluated against the material properties for aluminum-6061. The preliminary investigation will focus on a comparison of density, oxidation rates, water chemistry requirements, mechanical properties, thermal properties, and neutronic properties.

  4. Geometries, electronic states, and spectroscopic properties of nitrogen-doped fullerene fragment C10N2(II) and its ions.

    PubMed

    Zhu, Xiaolei

    2008-03-01

    The DFT(B3LYP)/6-31G(d)//CCSD(T)/6-31G(d) method is used to investigate the low-lying electronic states of C(10)N(2)(II) and its ions. Mulliken populations, leading configurations, bond orders, and compositions of molecular orbitals are employed to explore the nature of bonding in the electronic states of C(10)N(2)(II) and its ions. Electron affinity, ionization energy, binding energy of C(10)N(2)(II), and anion photoelectron spectra of C(10)N(2)(II)(-) are also estimated at the CCSD(T)/6-31G(d) level. On the other hand, the similarities and differences between C(10)N(2)(I) and C(10)N(2)(II) are compared and discussed.

  5. Wind speeds on extrasolar worlds

    NASA Astrophysics Data System (ADS)

    Allers, Katelyn; Biller, Beth; Vos, Johanna; Williams, Peter; Berger, Edo

    2016-08-01

    We propose for photometric monitoring observations of 2MASS J10475385+2124234 and WISE J112254.73+255021.5 using Spitzer/IRAC. 2MASS J1047+21 and WISE J1122+25 are late spectral type (T6.5 and T6) radio emitters and have measured radio periods of 1.77 hrs and 1.30 hrs, respectively. Our proposed observations will not only characterize the variability of the two coolest known radio emitters but also provide a unique opportunity to measure the first wind speeds for brown dwarfs. Spitzer is currently the only facility capable of the photometric stability, continuous observations and 4.5 micron sensitivity necessary for the success of our program.

  6. Evaluation of Johnson-Cook model constants for aluminum based particulate metal matrix composites

    NASA Astrophysics Data System (ADS)

    Hilfi, H.; Brar, N. S.

    1996-05-01

    High strain rate and high temperature response of three types of aluminum based particulate metal matrix ceramic composites is investigated by performing split Hopkinson pressure bar (SHPB) experiments. The composites are: NGP-2014 (15% SiC), NGT-6061 (15% SiC), and NGU-6061 (15% Al2O3), in which all the reinforcement materials are percentage by volume. Johnson-Cook constitutive model constants are evaluated from the high strain rate/high temperature data and implemented in a two dimensional finite element computer code (EPIC-2D) to simulate the penetration of an ogive nose tungsten projectile (23 grams) at a velocity 1.17 km/sec into the base 6061-T6 aluminum alloy and the composite NGU-6061. The simulated penetrations in the composite and in 6061-T6 aluminum agree with in 2%, in both materials, with the measured values.

  7. The Vibrio cholerae type VI secretion system employs diverse effector modules for intraspecific competition.

    PubMed

    Unterweger, Daniel; Miyata, Sarah T; Bachmann, Verena; Brooks, Teresa M; Mullins, Travis; Kostiuk, Benjamin; Provenzano, Daniele; Pukatzki, Stefan

    2014-04-01

    Vibrio cholerae is a Gram-negative bacterial pathogen that consists of over 200 serogroups with differing pathogenic potential. Only strains that express the virulence factors cholera toxin (CT) and toxin-coregulated pilus (TCP) are capable of pandemic spread of cholera diarrhoea. Regardless, all V. cholerae strains sequenced to date harbour genes for the type VI secretion system (T6SS) that translocates effectors into neighbouring eukaryotic and prokaryotic cells. Here we report that the effectors encoded within these conserved gene clusters differ widely among V. cholerae strains, and that immunity proteins encoded immediately downstream from the effector genes protect their host from neighbouring bacteria producing corresponding effectors. As a consequence, strains with matching effector-immunity gene sets can coexist, while strains with different sets compete against each other. Thus, the V. cholerae T6SS contributes to the competitive behaviour of this species.

  8. Discrete Abelian gauge symmetries and axions

    NASA Astrophysics Data System (ADS)

    Honecker, Gabriele; Staessens, Wieland

    2015-07-01

    We combine two popular extensions of beyond the Standard Model physics within the framework of intersecting D6-brane models: discrete ℤn symmetries and Peccei-Quinn axions. The underlying natural connection between both extensions is formed by the presence of massive U(1) gauge symmetries in D-brane model building. Global intersecting D6-brane models on toroidal orbifolds of the type T6/ℤ2N and T6/ℤ2 × ℤ2M with discrete torsion offer excellent playgrounds for realizing these extensions. A generation-dependent ℤ2 symmetry is identified in a global Pati-Salam model, while global left-right symmetric models give rise to supersymmetric realizations of the DFSZ axion model. In one class of the latter models, the axion as well as Standard Model particles carry a non-trivial ℤ3 charge.

  9. Dystrophic thoracic spine dislocation associated with type-1 neurofibromatosis: Case report and rationale for treatment

    PubMed Central

    Meneses-Quintero, David; Alvarado-Gómez, Fernando; Alcalá-Cerra, Gabriel

    2015-01-01

    The authors report a rare case of spontaneous dystrophic thoracic spine dislocation in a 14-year-old boy with neurofibromatosis type 1 (NF-1). Anteroposterior and lateral standing radiographs showed a dysplastic kyphoscoliotic deformity, with the thoracic kyphosis and scoliosis measuring 75° and 69°, respectively. Three-dimensional reconstruction after computed tomography demonstrated spondyloptosis at T5-T6 with overlapping of T5 over T6 and T7. The patient underwent circumferential fusion with anterior fibular strut grafting mechanically secured between the inferior and superior endplates of T5 and T7 followed by an instrumented posterior fusion from T2 to L1 and thoracoplasty. There was satisfactory resolution of the deformity with stabilization at the last follow-up evaluation. PMID:25972714

  10. Dystrophic thoracic spine dislocation associated with type-1 neurofibromatosis: Case report and rationale for treatment.

    PubMed

    Meneses-Quintero, David; Alvarado-Gómez, Fernando; Alcalá-Cerra, Gabriel

    2015-01-01

    The authors report a rare case of spontaneous dystrophic thoracic spine dislocation in a 14-year-old boy with neurofibromatosis type 1 (NF-1). Anteroposterior and lateral standing radiographs showed a dysplastic kyphoscoliotic deformity, with the thoracic kyphosis and scoliosis measuring 75° and 69°, respectively. Three-dimensional reconstruction after computed tomography demonstrated spondyloptosis at T5-T6 with overlapping of T5 over T6 and T7. The patient underwent circumferential fusion with anterior fibular strut grafting mechanically secured between the inferior and superior endplates of T5 and T7 followed by an instrumented posterior fusion from T2 to L1 and thoracoplasty. There was satisfactory resolution of the deformity with stabilization at the last follow-up evaluation.

  11. Trends of prevalent cancer incidences in the Aral-Syr Darya ecological area of Kazakhstan.

    PubMed

    Igissinov, Nurbek; Igissinov, Saginbek; Moore, Malcolm A; Shaidarov, Mazhit; Tereshkevich, Dmitriy; Bilyalova, Zarina; Igissinova, Gulnur; Nuralina, Indira; Kozhakhmetov, Saken

    2011-01-01

    The aim of this research was to examine the incidence of major forms of cancer in the Aral-Syr Darya ecological area of Kazakhstan. The present retrospective study of 11 years (1999-2009) was therefore conducted using descriptive and analytical methods. Incidence rates (crude and standardized) of 11 leading cancer sites were calculated and trends determined. The result of analysis demonstrated the most common neoplams in the study region to be esophageal cancer, carcinoma of lung, stomach cancer, and breast cancer. Trends in incidence of cancers under study were different, the most marked reduction in cancer of esophagus is established (T=-6.1%) and revealed the high increase in breast cancer (T=+6.7%). In the dynamics the trend of malignant disease in general tended to decrease (T=-0.5%).

  12. Interactions of satellite-speed helium atoms with satellite surfaces. 2: Energy distributions of reflected helium atoms

    NASA Technical Reports Server (NTRS)

    Liu, S. M.; Knuth, E. L.

    1976-01-01

    Energy transfer in collisions of satellite-speed (7,000 m/sec) helium atoms with a cleaned 6061-T6 satellite-type aluminum surface was investigated using the molecular-beam technique. The amount of energy transferred was determined from the measured energy of the molecular-beam and the measured spatial and energy distributions of the reflected atoms. Spatial distributions of helium atoms scattered from a 6061-T6 aluminum surface were measured. The scattering pattern exhibits a prominent backscattering, probably due to the gross surface roughness and/or the relative lattice softness of the aluminum surface. Energy distributions of reflected helium atoms from the same surface were measured for six different incidence angles. For each incidence angle, distributions were measured at approximately sixty scattering positions. At a given scattering position, the energy spectra of the reflected helium atoms and the background gas were obtained using the retarding-field energy analyzer.

  13. Osteopathic manipulative treatment in the management of biliary dyskinesia.

    PubMed

    Heineman, Katherine

    2014-02-01

    Biliary dyskinesia is a functional gastrointestinal disorder of the gallbladder and sphincter of Oddi. Diagnosis is made on the basis of symptoms of biliary colic in the absence of cholelithiasis and gallbladder inflammation. Palpatory findings of tissue texture changes at midthoracic levels (T6-T9) may correspond to visceral dysfunction related to the biliary system. Osteopathic manipulative treatment (OMT) of the T6-T9 segments can remove the feedback related to the somatic component, thereby affecting nociceptive facilitation at the spinal level and allowing the body to restore autonomic balance. Few reports in the current literature provide examples of treatment for patients with biliary dyskinesia using OMT. The author describes the case of a 51-year-old woman who presented with symptoms consistent with biliary dyskinesia. Her biliary colic completely resolved after OMT. Osteopathic evaluation and OMT should be considered a safe and effective option for conservative management of biliary dyskinesia.

  14. Generation of reactive oxygen species by lethal attacks from competing microbes

    PubMed Central

    Dong, Tao G.; Dong, Shiqi; Catalano, Christy; Moore, Richard; Liang, Xiaoye; Mekalanos, John J.

    2015-01-01

    Whether antibiotics induce the production of reactive oxygen species (ROS) that contribute to cell death is an important yet controversial topic. Here, we report that lethal attacks from bacterial and viral species also result in ROS production in target cells. Using soxS as an ROS reporter, we found soxS was highly induced in Escherichia coli exposed to various forms of attacks mediated by the type VI secretion system (T6SS), P1vir phage, and polymyxin B. Using a fluorescence ROS probe, we found enhanced ROS levels correlate with induced soxS in E. coli expressing a toxic T6SS antibacterial effector and in E. coli treated with P1vir phage or polymyxin B. We conclude that both contact-dependent and contact-independent interactions with aggressive competing bacterial species and viruses can induce production of ROS in E. coli target cells. PMID:25646446

  15. Late Occurrence of Cervicothoracic Ossification of Posterior Longitudinal Ligaments in a Surgically Treated Thoracic OPLL Patient

    PubMed Central

    Hyun, Seung-Jae; Hong, Seung-Chyul

    2010-01-01

    Ossification of the posterior longitudinal ligament (OPLL) in the thoracic spine is rare, even in the Far East. A 45-year-old female presented with a 4-month history of progressive motor weakness in the lower extremities, numbness below the midthoracic area, and spastic gait disturbance. Neuroradiological examinations revealed massive OPLLs at the T4-T6 levels with severe anterior compression of the spinal cord. Anterior decompressive corpectomies with bone grafts were performed from T4 to T6 using a trans-thoracic approach. After surgery, the patient made an uneventful recovery. However, eleven years after surgery, the patient developed recurrent lower extremity weakness and spastic gait disturbance. De novo OPLLs at the C6-T2 levels were responsible for the severe spinal cord compression on this occasion. After second surgery, paralysis in both legs was resolved. We present a rare case of late cervicothoracic OPLL in a patient surgically treated for thoracic OPLL. PMID:20157380

  16. Metal induced conformational changes in human insulin: crystal structures of Sr2+, Ni2+ and Cu2+ complexes of human insulin.

    PubMed

    Krishna, Nagampalli Raghavendra Sashi; Pattabhi, Vasantha; Rajan, S S

    2011-05-01

    Crystal structures of Sr(2+), Ni(2+) and Cu(2+) of human insulin complexes have been determined. The structures of Sr(2+) and Ni(2+) complexes are similar to Zn(2+) insulin and are in T6 conformation. (All the six monomers in the insulin hexamer are in Tensed conformation (T), which means the first eight residues of B-chain are in an extended conformation). Cu(2+) complex, though it assumes T6 conformation, has more structural differences due to lowering of crystal symmetry and space group shift from H3 (Hexagonal crystal system) to P3 (Trigonal crystal system) and a doubling of the c axis. 2Ni(2+) human insulin when compared to 4Ni(2+) Arg insulin suggests that terminal modifications may be responsible for additional metal binding. All the three metals have been shown to have a role in diabetes and hence may be therapeutically useful. PMID:21171943

  17. Axial-Load Fatigue Properties of 24S-T and 75S-T Aluminum Alloy as Determined in Several Laboratories

    NASA Technical Reports Server (NTRS)

    Grover, H J; Hyler, W S; Kuhn, Paul; Landers, Charles B; Howell, F M

    1954-01-01

    In the initial phase of a NACA program on fatigue research, axial-load tests on 24S-T3 and 75S-T6 aluminum-alloy sheet have been made at the Battelle Memorial Institute and at the Langley Aeronautical Laboratory of the National Advisory Committee for Aeronautics. The test specimens were polished and unnotched. The manufacturer of the material, the Aluminum Company of America, has made axial-load tests on 24S-T4 and 75S-T6 rod material. The test techniques used at the three laboratories are described in detail; the test results are presented and are compared with each other and with results obtained on unpolished sheet by the National Bureau of Standards. (author)

  18. Molecular analyses of the candidate tumor suppressor gene, PLAGL1, in benign and malignant salivary gland tumors.

    PubMed

    Enlund, Fredrik; Persson, Fredrik; Stenman, Göran

    2004-12-01

    Deletions affecting the long arm of chromosome 6 are a characteristic feature of all major subtypes of malignant salivary gland tumors. Moreover, a subgroup of adenoid cystic carcinomas have t(6;9)(q23-25;p21-24) translocations with breakpoints located within the commonly deleted region. Here we have examined the possible involvement of the candidate tumor suppressor gene, PLAGL1, in these deletions and translocations. Northern blot and fluorescence in situ hybridization (FISH) analyses of a series of 27 salivary gland tumors revealed no significant changes in the gene expression or rearrangements of PLAGL1. FISH analysis also demonstrated that the 6q translocation breakpoint in adenoid cystic carcinomas with t(6;9) is proximal to the PLAGL1 locus. Collectively, these results indicate that PLAGL1 is not likely to be the major target gene of the 6q rearrangements in salivary gland tumors.

  19. Intestinal microecology associated with fluoride resistance capability of the silkworm (Bombyx mori L.).

    PubMed

    Li, Guan-Nan; Xia, Xue-Juan; Tang, Wen-Chao; Zhu, Yong

    2016-08-01

    The silkworm (Bombyx mori L.) is an ideal model of Lepidoptera. However, the diversity and function of the intestinal microbiota in the gut of silkworm remain largely unknown. Changes in the intestinal microecology in fluoride-resistant strain T6 and fluoride-susceptible strain 734 of the silkworm in response to fluoride exposure were investigated. T6 and 734 were treated with 200 mg/kg fluoride (designated as T6-T and 734-T groups) and deionized water (designated as T6-C and 734-C groups). Culture-dependent approach revealed that the numbers of intestinal bacteria in the 734-T group significantly decreased compared with that in the 734-C group (4.8 ± 0.6 × 10(7) CFU/mL vs. 7.5 ± 0.7 × 10(7) CFU/mL; P < 0.05). Analyses of the intestinal content pH showed that the pH decreased in the 734-T group only. Additionally, SCFA concentrations significantly decreased in both treatment groups compared with the control groups. High-throughput sequencing indicated that the intestinal microbiota in the 734-T group was significantly more diverse than those in the other groups. The bacterial community was composed of two dominant groups (Firmicutes and Proteobacteria). Principal component analyses revealed a significant difference in the composition of the intestinal microbiota in the 734-T group compared with those in the other groups. Thaumarchaeota and Euryarchaeota were more abundant in the 734-T group, but they were less abundant in the other groups. This study enhances our understanding about the diversity and function of silkworm intestinal microbiota in response to fluoride exposure among silkworm strains with diverse resistance.

  20. Global geographic distribution of Trichinella species and genotypes.

    PubMed

    Feidas, Haralambos; Kouam, Marc K; Kantzoura, Vaia; Theodoropoulos, Georgios

    2014-08-01

    Maximum entropy ecological niche modeling was utilized to describe the global geographic distribution of Trichinella species and genotypes and to assess their invasive risk in new areas other than the ones currently known. Also, space-time scan statistic was utilized to identify global spatiotemporal clusters of infection. A database containing 3209 records for 12 species and genotypes identified at the International Trichinella Reference Center (ITRC) as well as climate, elevation, and land cover data extracted from various databases were used. Ecological niche modeling implemented in the Maxent program indicated new potential ranges for T. spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4), T. murrelli (T5), T6, T. papuae (T10), and T. zimbabwensis (T11). The area under the curve values for the test data of the models ranged from 0.901 to 0.998, indicating that the models were very good to excellent. The most important bioclimatic factor in modeling the ranges for T. spiralis (T1), T. nativa (T2), T. britovi (T3), T6, and T. zimbabwensis (T11) was temperature, for T. pseudospiralis (T4) and T. papuae (T10) was precipitation, and for T. murrelli (T5) was land cover. T. spiralis (T1), T. britovi (T3), and T. pseudospiralis (T4) had the same primary land cover which was "Grass Crops". The primary land covers were "Conifer Boreal Forest" for T. nativa (T2), "Cool Fields and Woods" for T. murrelli (T5), "Upland Tundra" for T6, "Tropical Rainforest" for T. papuae (T10), and "Crops and Town" for T. zimbabwensis (T11). The scan statistic analyses revealed the presence of significant spatiotemporal clusters (p<0.05) for T. spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4), T. murrelli (T5), T6, and T. nelsoni (T7). No significant clusters were found for T. papuae (T10) and T. zimbabwensis (T11).

  1. Study on antifibrotic effects of curcumin in rat hepatic stellate cells.

    PubMed

    Lin, Yun-Lian; Lin, Chia-Yu; Chi, Chin-Wen; Huang, Yi-Tsau

    2009-07-01

    Suppression of activation or fibrogenesis and induction of apoptosis, in hepatic stellate cells (HSCs) have been proposed as therapeutic strategies against liver fibrosis. Curcumin, an active compound isolated from yellow curry pigment of turmeric (Curcuma longa Linn), has been demonstrated to be an effective anti-inflammatory and antioxidant compound. In this study, we investigated the in vitro antifibrogenic effects of curcumin on HSCs at the concentration range of (1-40 microM). A cell line of rat HSCs (HSC-T6) was stimulated with transforming growth factor-beta1 (TGF-beta1). The inhibitory effects of curcumin (1.25 approximately 10 microM) on fibrosis-related markers including alpha-smooth muscle actin (alpha-SMA) and collagen were assessed. In addition, the induction effects of curcumin (20 approximately 40 microM) on apoptosis in HSC-T6 cells were also assessed by Hoechst and propidium iodide stains. Curcumin (1.25 approximately 10 microM) concentration-dependently suppressed TGF-beta1-induced alpha-SMA expression and collagen deposition in HSC-T6 cells, without cytotoxicity. Whereas, higher concentrations of curcumin (20 approximately 40 microM) induced cell apoptosis and cytochrome c release in HSC-T6 cells. Our results suggest that curcumin exerted antifibrotic effects, possibly through two different mechanisms depending on its concentrations. At lower concentrations (1.25 approximately 10 microM), curcumin exerted antifibrogenic effects, whereas at higher concentrations (20 approximately 40 microM), curcumin exerted induction of apoptosis in HSCs. PMID:19152370

  2. Indole-3-carbinol enhances the resolution of rat liver fibrosis and stimulates hepatic stellate cell apoptosis by blocking the inhibitor of κB kinase α/inhibitor of κB-α/nuclear factor-κB pathway.

    PubMed

    Ping, Jie; Gao, Ai-mei; Qin, Hai-quan; Wei, Xiao-ning; Bai, Jing; Liu, Lian; Li, Xiao-hai; Li, Rui-wen; Ao, Ying; Wang, Hui

    2011-11-01

    Hepatic stellate cells (HSC) play a pivotal role in liver fibrosis, and the clearance of activated HSC by apoptosis is associated with the resolution of liver fibrosis. The development of strategies that promote this process in a selective way is therefore important. We evaluated the effects of indole-3-carbinol (I3C), a nutritional component derived from vegetables from the Brassica family, on liver fibrosis and HSC apoptosis. The in vivo therapeutic effects of I3C were monitored in three rat models of liver fibrosis induced by porcine serum, bile duct ligation, or multiple hepatotoxic factors, and its proapoptotic effect and molecular mechanism were studied in vitro in HSC-T6, a rat HSC line. The results showed that I3C treatment significantly reduced the number of activated HSC in the livers of rats with liver fibrosis. In histopathology, I3C reduced hepatocyte degeneration and necrosis, accelerated collagen degradation, and promoted the reversal of liver fibrosis. I3C prescribed to HSC-T6 resulted in morphologic alterations typical of apoptosis and DNA cleavage to a nucleosomal ladder. Moreover, I3C significantly increased the HSC-T6 apoptosis rate and the expression ratio of Bax to Bcl-2. High-throughput protein array analysis indicated that the tumor necrosis factor-α/nuclear factor-κB (NF-κB) signal pathway participated in I3C-induced HSC-T6 apoptosis. Western blot and electrophoretic mobility-shift assay confirmed that I3C inhibited the phosphorylation of inhibitor of κB kinase α and inhibitor of κB-α and NF-κB DNA binding activity. In conclusion, I3C could promote the reverse process of liver fibrosis in vivo and induce apoptosis of activated HSC in vitro, which indicates the use of I3C as a potential therapeutic agent in liver fibrosis treatment.

  3. Phase I trial of sunitinib and temsirolimus in metastatic renal cell carcinoma

    PubMed Central

    Campbell, Matthew T.; Millikan, Randall E.; Altinmakas, Emre; Xiao, Lianchun; Wen, Sin Jen; Siefker-Radtke, Arlene O.; Aparicio, Ana; Corn, Paul G.; Tannir, Nizar M.

    2015-01-01

    Purpose Preclinical data suggest that anti-VEGF agents plus mTOR inhibitors yield synergistic antitumor effects. A phase I trial using a 3+3 dose escalation design of sunitinib (S) plus temsirolimus (T) was stopped after the first dose pair led to 2 of 3 patients experiencing dose-limiting toxicity (DLT). Patients and Methods To explore multiple potential dosing pairs of S and T a 2 stage outcome-adaptive Bayesian dose-finding method was designed. The primary objective was to find the maximum tolerated doses (MTD) of S and T in patients with advanced RCC. A 3-wk treatment cycle consisted of daily S, 2 wks on, 1 wk off, plus weekly T. Results Twenty patients received study drugs; median number of prior therapies 1. Number of patients (S and T doses in mg): 2 (S12.5, T6), 1 (S25, T12.5), 1 (S12.5, T8), 8 (S12.5 alt 25, T9), 2 (S25, T6), 2 (S25 alt 37.5, T6), 2 (S37.5, T6), 2 (S37.5, T8). Six patients required dose reduction, 3 due to grade 3 stomatitis, 2 due to grade 3 thrombocytopenia; mean number of cycles 6.6 ± 5.3, mean time on study 159±120 days. One patient experienced DLT in cycle 1 and was non-evaluable, 1 had a partial response, 16 had stable disease, and 2 had progressive disease (PD) as best response. There were 21 grade 3/4 adverse events but no treatment-related deaths. Conclusions The MTD of S and T were not determined because of premature trial closure. S 37.5 mg/d, 2 wks on, 1 wk off, plus T 8–10 mg weekly are close to MTD. PMID:25465491

  4. [Effects of nitrogen management on yield, quality, nitrogen accumulation and its transportation of watermelon in gravel-mulched field].

    PubMed

    Ma, Zhong-ming; Du, Shao-ping; Xue, Liang

    2015-11-01

    The effects of nitrogen management on yield, quality, nitrogen and dry matter accumulation and transportation of watermelon in sand field were studied based on a field experiment. The results showed that too low or too high basal nitrogen fertilzation was unfavorable to seedling growth of watermelon in sand field, and no nitrogen application at vine extension or fruiting stages limited the formation of 'source' or 'sink'. At the same nitrogen rate, compared with the traditional T1 treatment (30% basal N fertilizer + 70% N fertilizer in vine extension), the nitrogen and dry matter accumulation of vegetative organs of T4 treatment (30% basal N fertilizer + 30% N fertilizer in vine extension + 40% N fertilizer in fruiting) and T6 treatment (100% basal N fertilizer + NAM) were reduced significantly, but the nitrogen and dry matter accumulation of fruit were increased significantly in the flushing period. The nitrogen transportation ratio and nitrogen contribution ratio of T4 were 33.6% and 12.0%, respectively. Compared to T1, the nitrogen harvest index, nitrogen fertilizer partial factor productivity and nitrogen fertilizer recovery efficiency of T4 and T6 treatments increased by 14.1% and 12.7%, 11.6% and 12.5%, 5.3% and 8.7%, respectively, and yield of watermelon increased by 11.6% and 12.5%, the soluble sugar, effective acid, the ratio of sugar and acid, Vc content increased by 16.5% and 11.7%, 4.5% and 2.8%, 19.4% and 13.4%, 35.6% and 19.0%, respectively. Therefore, T4 and T6 treatments were the optimal nitrogen fertilizer management mode which could not only achieve high yield and quality but also obtain high nitrogen fertilizer use efficiency in sand field. T6 treatment was the best nitrogen fertilizer management mode considering reduction of fertilizing labor intensity and extending service time of gravel-mulched field.

  5. Dynamic properties of the upper thoracic spine-pectoral girdle (UTS-PG) system and corresponding kinematics in PMHS sled tests.

    PubMed

    Stammen, Jason A; Herriott, Rodney; Kang, Yun-Seok; Dupaix, Rebecca; Bolte, John

    2012-10-01

    Anthropomorphic test devices (ATDs) should accurately depict head kinematics in crash tests, and thoracic spine properties have been demonstrated to affect those kinematics. To investigate the relationships between thoracic spine system dynamics and upper thoracic kinematics in crash-level scenarios, three adult post-mortem human subjects (PMHS) were tested in both Isolated Segment Manipulation (ISM) and sled configurations. In frontal sled tests, the T6-T8 vertebrae of the PMHS were coupled through a novel fixation technique to a rigid seat to directly measure thoracic spine loading. Mid-thoracic spine and belt loads along with head, spine, and pectoral girdle (PG) displacements were measured in 12 sled tests conducted with the three PMHS (3-pt lap-shoulder belted/unbelted at velocities from 3.8 - 7.0 m/s applied directly through T6-T8). The sled pulse, ISM- derived characteristic properties of that PMHS, and externally applied forces due to head-neck inertia and shoulder belt constraint were used to predict kinematic time histories of the T1-T6 spine segment. The experimental impulse applied to the upper thorax was normalized to be consistent with a T6 force/sled acceleration sinusoidal profile, and the result was an improvement in the prediction of T3 X-axis displacements with ISM properties. Differences between experimental and model-predicted displacement-time history increases were quantified with respect to speed. These discrepancies were attributed to the lack of rotational inertia of the head-neck late in the event as well as restricted kyphosis and viscoelasticity of spine constitutive structures through costovertebral interactions and mid-spine fixation. The results indicate that system dynamic properties from sub-injurious ISM testing could be useful for characterizing forward trajectories of the upper thoracic spine in higher energy crash simulations, leading to improved biofidelity for both ATDs and finite element models.

  6. 40 CFR 415.82 - Effluent limitations guidelines representing the degree of effluent reduction attainable by the...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... technology currently available (BPT). Except as provided in 40 CFR 125.30 through 125.32, any existing point... SOURCE CATEGORY Hydrofluoric Acid Production Subcategory § 415.82 Effluent limitations guidelines... product TSS 11.0 5.3 Fluoride (T) 6.1 2.9 Nickel (T) k0.036 0.011 Zinc (T) 0.12 0.036 pH (1) (1) 1...

  7. A Disordered Region in the EvpP Protein from the Type VI Secretion System of Edwardsiella tarda is Essential for EvpC Binding

    PubMed Central

    Hu, Wentao; Anand, Ganesh; Sivaraman, J.; Leung, Ka Yin; Mok, Yu-Keung

    2014-01-01

    The type VI secretion system (T6SS) of pathogenic bacteria plays important roles in both virulence and inter-bacterial competitions. The effectors of T6SS are presumed to be transported either by attaching to the tip protein or by interacting with HcpI (haemolysin corregulated protein 1). In Edwardsiella tarda PPD130/91, the T6SS secreted protein EvpP (E. tarda virulent protein P) is found to be essential for virulence and directly interacts with EvpC (Hcp-like), suggesting that it could be a potential effector. Using limited protease digestion, nuclear magnetic resonance heteronuclear Nuclear Overhauser Effects, and hydrogen-deuterium exchange mass spectrometry, we confirmed that the dimeric EvpP (40 kDa) contains a substantial proportion (40%) of disordered regions but still maintains an ordered and folded core domain. We show that an N-terminal, 10-kDa, protease-resistant fragment in EvpP connects to a shorter, 4-kDa protease-resistant fragment through a highly flexible region, which is followed by another disordered region at the C-terminus. Within this C-terminal disordered region, residues Pro143 to Ile168 are essential for its interaction with EvpC. Unlike the highly unfolded T3SS effector, which has a lower molecular weight and is maintained in an unfolded conformation with a dedicated chaperone, the T6SS effector seems to be relatively larger, folded but partially disordered and uses HcpI as a chaperone. PMID:25401506

  8. Neferine inhibits cultured hepatic stellate cell activation and facilitates apoptosis: A possible molecular mechanism.

    PubMed

    Ding, Hui; Shi, Jinghong; Wang, Ying; Guo, Jia; Zhao, Juhui; Dong, Lei

    2011-01-10

    Neferine is a major alkaloid component of "Lian Zi Xin", embryos of the seeds of Nelumbo nucifera Gaertner, Nymphaeaceae. Previous studies have shown that neferine has an inhibitory effect on pulmonary fibrosis through its anti-inflammatory and anti-oxidative activities and inhibition of cytokines and NF-κB. However, it is unknown whether neferine also has an inhibitory effect on liver fibrosis through inhibition of TGF-β1 and collagen I and facilitation of apoptosis of hepatic stellate cells. This study examined the effects of neferine on cultured hepatic stellate (HSC-T6) cells and explored its possible action mechanisms by means of MTT assay, enzyme-linked immunosorbent assay, flow-cytometric annexin V-PI assay and Hoechst 33258 staining, as well as real-time PCR and western blotting. The results showed that neferine administration (2, 4, 6, 8 and 10μmol/l) significantly decreased the TGF-β1 and collagen I produced in HSC-T6 cells, and increased the HSC-T6 cell apoptosis in a dose-dependent manner. Neferine treatment for 48h at concentrations of 6 and 10μmol/l significantly increased Bax and caspase 3 mRNAs and proteins, and reduced Bcl2 and alpha-smooth muscle actin (α-SMA) mRNAs and proteins. Our data indicate that neferine efficiently inhibits cultured HSC-T6 cell activation and induces apoptosis by increasing Bax and caspase 3 expression via the mitochondrial pathway.

  9. Effect of workpiece edge angle on radius produced by vibratory finishing

    SciTech Connect

    Gillespie, L.K.

    1981-06-01

    An investigation was conducted to determine the effect of the included edge angle upon the radius produced by vibratory finishing of small precision parts. Workpiece materials studied included phosphor bronze, 6061-T6 aluminum, and 303Se stainless steel. Edges having included angles smaller than 90/sup 0/ require finishing cycles up to 20 times longer than those having angles greater than 90/sup 0/. The effect of the edge angle can be easily calculated.

  10. Trehalose-6-phosphate and SnRK1 kinases in plant development and signaling: the emerging picture

    PubMed Central

    Tsai, Allen Y.-L.; Gazzarrini, Sonia

    2014-01-01

    Carbohydrates, or sugars, regulate various aspects of plant growth through modulation of cell division and expansion. Besides playing essential roles as sources of energy for growth and as structural components of cells, carbohydrates also regulate the timing of expression of developmental programs. The disaccharide trehalose is used as an energy source, as a storage and transport molecule for glucose, and as a stress-responsive compound important for cellular protection during stress in all kingdoms. Trehalose, however, is found in very low amounts in most plants, pointing to a signaling over metabolic role for this non-reducing disaccharide. In the last decade, trehalose-6-phosphate (T6P), an intermediate in trehalose metabolism, has been shown to regulate embryonic and vegetative development, flowering time, meristem determinacy, and cell fate specification in plants. T6P acts as a global regulator of metabolism and transcription promoting plant growth and triggering developmental phase transitions in response to sugar availability. Among the T6P targets are members of the Sucrose-non-fermenting1-related kinase1 (SnRK1) family, which are sensors of energy availability and inhibit plant growth and development during metabolic stress to maintain energy homeostasis. In this review, we will discuss the opposite roles of the sugar metabolite T6P and the SnRK1 kinases in the regulation of developmental phase transitions in response to carbohydrate levels. We will focus on how these two global regulators of metabolic processes integrate environmental cues and interact with hormonal signaling pathways to modulate plant development. PMID:24744765

  11. Location of Si vacancies and [Ti(OSi)4] and [Ti(OSi)3OH] sites in the MFI framework: a large cluster and full ab initio study.

    PubMed

    Yuan, Shuping; Si, Hongzong; Fu, Aiping; Chu, Tianshu; Tian, Fenghui; Duan, Yun-Bo; Wang, Jianguo

    2011-02-10

    Titanium silicalite-1 (TS-1) is an important catalyst for selective oxidation reactions. However, the nature and structure of the active sites and the mechanistic details of the catalytic reactions over TS-1 have not been well-understood, leaving a continuous debate on the genesis of active sites on the TS-1 surface in the literature. In this work, the location of Si vacancies and [Ti(OSi)(4)] and [Ti(OSi)(3)OH] sites in the MFI (Framework Type Code of ZSM-5 (Zeolite Socony Mobile-Five)) framework has been studied using a full ab initio method with 40T clusters with a Si:Ti molar ratio of 39:1. It was shown that the former four energetically favorable sites for Si vacancies are T6, T12, T4, and T8 and for Ti centers of [Ti(OSi)(4)] are T10, T4, T8 and T11, being partially the same sites. Whether by replacing Si vacancies or substituting the fully coordinated Si sites, the most preferential site for Ti is T10, which indicates that the insertion mechanism does not affect the favorable sites of Ti in the MFI lattice. For the defective [Ti(OSi)(3)OH] sites, it was found that the Si vacancy at T6 with a Ti at its neighboring T9 site (T6-def-T9-Ti pair) is the most energetically favorable one, followed by a T6-def-T5-Ti pair with a small energy gap. These findings are significant to elucidate the nature of the active sites and the mechanism of reactions catalyzed by TS-1 and to design the TS-1 catalyst.

  12. 40 CFR 81.313 - Idaho.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... CFR Sections Affected which appears in the Finding Aids section of the printed volume and at www.fdsys...: (Pocatello) State Lands-Portneuf Valley Area: T.5S, R.34E Sections 25-36 T.5S, R.35E Section 31 T.6S, R.34E..., part of: (Pocatello) 11/15/90 Nonattainment 11/15/90 Moderate Fort Hall Indian Reservation: T.5S,...

  13. 40 CFR 81.313 - Idaho.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    .... Editorial Note: For Federal Register citations affecting § 81.313 see the List of CFR Sections Affected...: (Pocatello) State Lands-Portneuf Valley Area: T.5S, R.34E Sections 25-36 T.5S, R.35E Section 31 T.6S, R.34E..., part of: (Pocatello) 11/15/90 Nonattainment 11/15/90 Moderate Fort Hall Indian Reservation: T.5S,...

  14. 40 CFR 81.313 - Idaho.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Register citations affecting § 81.313 see the List of CFR Sections Affected, which appears in the Finding...: (Pocatello) State Lands-Portneuf Valley Area: T.5S, R.34E Sections 25-36 T.5S, R.35E Section 31 T.6S, R.34E..., part of: (Pocatello) 11/15/90 Nonattainment 11/15/90 Moderate Fort Hall Indian Reservation: T.5S,...

  15. 40 CFR 81.313 - Idaho.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... CFR Sections Affected which appears in the Finding Aids section of the printed volume and on GPO...: (Pocatello) State Lands-Portneuf Valley Area: T.5S, R.34E Sections 25-36 T.5S, R.35E Section 31 T.6S, R.34E..., part of: (Pocatello) 11/15/90 Nonattainment 11/15/90 Moderate Fort Hall Indian Reservation: T.5S,...

  16. Electron Positron Proton Spectrometer for use at Laboratory for Laser Energetics

    SciTech Connect

    Ayers, S L

    2010-04-07

    The Electron Positron Proton Spectrometer (EPPS) is mounted in a TIM (Ten-Inch Manipulator) system on the Omega-60 or Omega-EP laser facilities at the University of Rochester, Laboratory for Laser Energetics (LLE), when in use, see Fig. 1. The Spectrometer assembly, shown in Fig. 2, is constructed of a steel box containing magnets, surrounded by Lead 6% Antimony shielding with SS threaded insert, sitting on an Aluminum 6061-T6 plate.

  17. LDEF (Prelaunch)

    NASA Technical Reports Server (NTRS)

    1983-01-01

    LDEF (Prelaunch) The LDEF structure is shown mounted on the LDEF Assembly and Transport System in Building 1250 at the Langley Research Center (LaRC), Hampton Virginia, during pre-ship wiring checks. The structure is a welded / bolted assembly fab- ricated from aluminum alloy 6061-T6 extrusions, forgings, tubing and plate materials and assembled with stainless steel fasteners. The eight (8) internal tubular diagonals between the center ring structure and the two end frames provide torsional stiffness to the structure.

  18. Novel matrine derivative MD-1 attenuates hepatic fibrosis by inhibiting EGFR activation of hepatic stellate cells.

    PubMed

    Feng, Yi; Ying, Hai-Yan; Qu, Ying; Cai, Xiao-Bo; Xu, Ming-Yi; Lu, Lun-Gen

    2016-09-01

    Matrine (MT), the effective component of Sophora flavescens Ait, has been shown to have anti-inflammation, immune-suppressive, anti-tumor, and anti-hepatic fibrosis activities. However, the pharmacological effects of MT still need to be strengthened due to its relatively low efficacy and short half-life. In the present study, we report a more effective thio derivative of MT, MD-1, and its inhibitory effects on the activation of hepatic stellate cells (HSCs) in both cell culture and animal models. Cytological experiments showed that MD-1 can inhibit the proliferation of HSC-T6 cells with a half-maximal inhibitory concentration (IC50) of 62 μmol/L. In addition, MD-1 more strongly inhibits the migration of HSC-T6 cells compared to MT and can more effectively induce G0/G1 arrest and apoptosis. Investigating the biological mechanisms underlying anti-hepatic fibrosis in the presence of MD-1, we found that MD-1 can bind the epidermal growth factor receptor (EGFR) on the surface of HSC-T6 cells, which can further inhibit the phosphorylation of EGFR and its downstream protein kinase B (Akt), resulting in decreased expression of cyclin D1 and eventual inhibition of the activation of HSC-T6 cells. Furthermore, in rats with dimethylnitrosamine (DMN)-induced hepatic fibrosis, MD-1 slowed the development and progression of hepatic fibrosis, protecting hepatic parenchymal cells and improving hepatic functions. Therefore, MD-1 is a potential drug for anti-hepatic fibrosis.

  19. Neferine inhibits cultured hepatic stellate cell activation and facilitates apoptosis: A possible molecular mechanism.

    PubMed

    Ding, Hui; Shi, Jinghong; Wang, Ying; Guo, Jia; Zhao, Juhui; Dong, Lei

    2011-01-10

    Neferine is a major alkaloid component of "Lian Zi Xin", embryos of the seeds of Nelumbo nucifera Gaertner, Nymphaeaceae. Previous studies have shown that neferine has an inhibitory effect on pulmonary fibrosis through its anti-inflammatory and anti-oxidative activities and inhibition of cytokines and NF-κB. However, it is unknown whether neferine also has an inhibitory effect on liver fibrosis through inhibition of TGF-β1 and collagen I and facilitation of apoptosis of hepatic stellate cells. This study examined the effects of neferine on cultured hepatic stellate (HSC-T6) cells and explored its possible action mechanisms by means of MTT assay, enzyme-linked immunosorbent assay, flow-cytometric annexin V-PI assay and Hoechst 33258 staining, as well as real-time PCR and western blotting. The results showed that neferine administration (2, 4, 6, 8 and 10μmol/l) significantly decreased the TGF-β1 and collagen I produced in HSC-T6 cells, and increased the HSC-T6 cell apoptosis in a dose-dependent manner. Neferine treatment for 48h at concentrations of 6 and 10μmol/l significantly increased Bax and caspase 3 mRNAs and proteins, and reduced Bcl2 and alpha-smooth muscle actin (α-SMA) mRNAs and proteins. Our data indicate that neferine efficiently inhibits cultured HSC-T6 cell activation and induces apoptosis by increasing Bax and caspase 3 expression via the mitochondrial pathway. PMID:20969858

  20. QCD equation of state from the lattice

    SciTech Connect

    Borsanyi, Sz.; Jakovac, A.; Ratti, C.; Szabo, K. K.; Endro''di, G.; Katz, S. D.; Fodor, Z.; Krieg, S.

    2011-05-23

    We calculate the QCD equation of state with 2+1 staggered lattice flavors and a physical pion mass. We present precision data on the trace anomaly and pressure based on simulations at N{sub t} = 6,8 and 10. These results are confirmed by N{sub t} = 12 simulations at three temperatures. Detailed results can be found in [arXiv:1007.2580v2].

  1. 27 CFR 9.85 - Willow Creek.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... where the 1,000-foot contour line intersects Kirkham Creek (directly north of section 19, T.7 N./R.5E.), beginning in a southerly direction, the boundary line the 1,000-foot contour line to; (1) The point of intersection between the 1,000-foot contour line and the north section line of section 27, T.6N./R.5E.;...

  2. 27 CFR 9.85 - Willow Creek.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... where the 1,000-foot contour line intersects Kirkham Creek (directly north of section 19, T.7 N./R.5E.), beginning in a southerly direction, the boundary line the 1,000-foot contour line to; (1) The point of intersection between the 1,000-foot contour line and the north section line of section 27, T.6N./R.5E.;...

  3. 27 CFR 9.91 - Walla Walla Valley.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... point is on the Walla Walla quadrangle map, in T8N/37E, at the point where the 2,000 foot contour line..., Washington (on maps measured in metric units, this elevation is between the 600 and 650 meter contour lines... moving onto the Pendleton U.S.G.S. map, where it meets the 450 m contour line in T6N/R32E, near...

  4. 27 CFR 9.85 - Willow Creek.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... where the 1,000-foot contour line intersects Kirkham Creek (directly north of section 19, T.7 N./R.5E.), beginning in a southerly direction, the boundary line the 1,000-foot contour line to; (1) The point of intersection between the 1,000-foot contour line and the north section line of section 27, T.6N./R.5E.;...

  5. 27 CFR 9.91 - Walla Walla Valley.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... point is on the Walla Walla quadrangle map, in T8N/37E, at the point where the 2,000 foot contour line..., Washington (on maps measured in metric units, this elevation is between the 600 and 650 meter contour lines... moving onto the Pendleton U.S.G.S. map, where it meets the 450 m contour line in T6N/R32E, near...

  6. 27 CFR 9.91 - Walla Walla Valley.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... point is on the Walla Walla quadrangle map, in T8N/37E, at the point where the 2,000 foot contour line..., Washington (on maps measured in metric units, this elevation is between the 600 and 650 meter contour lines... moving onto the Pendleton U.S.G.S. map, where it meets the 450 m contour line in T6N/R32E, near...

  7. 27 CFR 9.91 - Walla Walla Valley.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... point is on the Walla Walla quadrangle map, in T8N/37E, at the point where the 2,000 foot contour line..., Washington (on maps measured in metric units, this elevation is between the 600 and 650 meter contour lines... moving onto the Pendleton U.S.G.S. map, where it meets the 450 m contour line in T6N/R32E, near...

  8. 27 CFR 9.85 - Willow Creek.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... where the 1,000-foot contour line intersects Kirkham Creek (directly north of section 19, T.7 N./R.5E.), beginning in a southerly direction, the boundary line the 1,000-foot contour line to; (1) The point of intersection between the 1,000-foot contour line and the north section line of section 27, T.6N./R.5E.;...

  9. 27 CFR 9.85 - Willow Creek.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... where the 1,000-foot contour line intersects Kirkham Creek (directly north of section 19, T.7 N./R.5E.), beginning in a southerly direction, the boundary line the 1,000-foot contour line to; (1) The point of intersection between the 1,000-foot contour line and the north section line of section 27, T.6N./R.5E.;...

  10. 27 CFR 9.91 - Walla Walla Valley.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... point is on the Walla Walla quadrangle map, in T8N/37E, at the point where the 2,000 foot contour line..., Washington (on maps measured in metric units, this elevation is between the 600 and 650 meter contour lines... moving onto the Pendleton U.S.G.S. map, where it meets the 450 m contour line in T6N/R32E, near...

  11. Effects of Post-Weld Heat Treatment on the Mechanical Properties of Similar- and Dissimilar-Alloy Friction Stir Welded Blanks

    SciTech Connect

    Zadpoor, Amir Abbas; Sinke, Jos

    2011-01-17

    Friction stir welding is a solid state joining process with relatively low welding temperatures. Nevertheless, the mechanical properties of friction stir welded blanks are degraded after welding. Indeed, both strength and ductility of the welds are decreased after welding. Often, the resulting friction stir welded blanks need to be formed to their final structural shape. Therefore, the formability of friction stir welded blanks is of primary importance in the manufacturing of structural parts. This paper studies how the mechanical properties and particularly formability of friction stir welded blanks can be improved by applying a post weld heat treatment. Two aluminum alloys from 2000 and 7000 series, namely 2024-T3 and 7075-T6, are selected for the study. The sheet thickness of both materials is 2,0 mm. The selected alloys are welded in three configurations: 2024-T3 and 2024-T3, 7075-T6 and 7075-T6, and 2024-T3 and 7075-T6. The resulting welds are naturally aged for a few months. Three sets of standard dog bone shape tensile test specimens are then machined from the welds. The first set of the specimens is tested without any heat treatment. The second set of the specimens is solution heat treated and quenched before testing. The third set of the specimens is solution heat treated, quenched, and naturally aged for a week before testing. The mechanical properties of the three different sets of specimens are compared with each other. It is shown that careful selection of post weld heat-treatment can greatly improve the formability of friction stir welded blanks.

  12. The PP-motif in luminal loop 2 of ZnT transporters plays a pivotal role in TNAP activation.

    PubMed

    Fujimoto, Shigeyuki; Tsuji, Tokuji; Fujiwara, Takashi; Takeda, Taka-Aki; Merriman, Chengfeng; Fukunaka, Ayako; Nishito, Yukina; Fu, Dax; Hoch, Eitan; Sekler, Israel; Fukue, Kazuhisa; Miyamae, Yusaku; Masuda, Seiji; Nagao, Masaya; Kambe, Taiho

    2016-09-01

    Secretory and membrane-bound zinc-requiring enzymes are thought to be activated by binding zinc in the early secretory pathway. One such enzyme, tissue-non-specific alkaline phosphatase (TNAP), is activated through a two-step mechanism, via protein stabilization and subsequent enzyme activation through metalation, by ZnT5-ZnT6 heterodimers or ZnT7 homodimers. However, little is known about the molecular basis underlying the activation process. In the present study, we found that the di-proline motif (PP-motif) in luminal loop 2 of ZnT5 and ZnT7 is important for TNAP activation. TNAP activity was significantly reduced in cells lacking ZnT5-ZnT6 heterodimers and ZnT7 homodimers [triple knockout (TKO) cells]. The decreased TNAP activity was restored by expressing hZnT5 with hZnT6 or hZnT7, but significantly less so (almost 90% less) by expressing mutants thereof in which the PP-motif was mutated to alanine (PP-AA). In TKO cells, overexpressed hTNAP was not completely activated, and it was converted less efficiently into the holo form by expressing a PP-AA mutant of hZnT5 with hZnT6, whose defects were not restored by zinc supplementation. The zinc transport activity of hZnT7 was not significantly impaired by the PP-AA mutation, indicating that the PP-motif is involved in the TNAP maturation process, although it does not control zinc transport activity. The PP-motif is highly conserved in ZnT5 and ZnT7 orthologues, and its importance for TNAP activation is conserved in the Caenorhabditis elegans hZnT5 orthologue CDF5. These results provide novel molecular insights into the TNAP activation process in the early secretory pathway. PMID:27303047

  13. An invasive and low virulent Edwardsiella tarda esrB mutant promising as live attenuated vaccine in aquaculture.

    PubMed

    Yang, Weizheng; Wang, Lixia; Zhang, Lingzhi; Qu, Jiangbo; Wang, Qiyao; Zhang, Yuanxing

    2015-02-01

    Edwardsiella tarda is a leading fish pathogen haunting worldwide aquaculture industry. In E. tarda, two-component system EsrA-EsrB positively regulates type III and VI secretion systems (T3SS and T6SS) and negatively regulates hemolysin EthA, which has been demonstrated to be essential for the invasion processes in fish. In order to develop a live attenuated vaccine (LAV) with high invasiveness to be practically and economically used as immersion-administered vaccine in aquaculture, here, we generated a random mutation library of esrB sequences by error-prone PCR and introduced them into the E. tarda esrB deletion mutant. The mutant YWZ47 with significantly increased hemolytic activity and low T3SS and T6SS secretion was screened. Phenotypes including extracellular protein profiles, invasion in macrophages, lethality toward fish, and infection kinetics were investigated in the wild-type strain EIB202 and the mutants ΔesrB, ΔT3SS, ΔT6SS, ΔT3SS/ΔT6SS, and YWZ47. Compared to the documented LAV strain ΔesrB, YWZ47 showed higher invasive capability and low in vivo virulence toward fish. Significantly higher relative percent survival (RPS) could be generated in turbot (Scophthalmus maximus) against the challenge of the wild-type EIB202 when inoculated through immersion route, and the RPS was comparable with that of ΔesrB through intraperitoneal (i.p.) injection inoculation. Two mutated points, K167M and H197L, were found by sequence analysis of EsrBYWZ47 variant. These structural modifications underpin the variations in the regulatory functions of the mutant and wild-type EsrB. This study promoted understanding of virulence regulation by EsrB in E. tarda and presented a promising candidate of invasive attenuated vaccine used in aquaculture industries. PMID:25431010

  14. Histone H3 Phosphorylation in GBM: a New Rational to Guide the Use of Kinase Inhibitors in anti-GBM Therapy

    PubMed Central

    Pacaud, Romain; Cheray, Mathilde; Nadaradjane, Arulraj; Vallette, François M.; Cartron, Pierre-François

    2015-01-01

    Histones post-translational modifications (PTMs) are crucial components of diverse processes that modulate chromatin. Among the histones PTMs, the histones phosphorylation appears such crucial since it plays a significant role into DNA repair structure, transcription and chromatin compaction during cell division and apoptosis. However, little is known about the prognostic value of the histone phosphorylation in human cancer. This point could be considerate such as an important gap in anti-cancer therapy since the use of adequate kinase inhibitors could remedy to the aberrant histone phosphorylation associated with a poor prognosis factor. To remedy at this situation, we analyzed the phosphorylation level of histone H3 at the residues T3, T6, S10, S28, Y41 and T45 in a collection of 42 glioblastoma multiformes (GBM). Our data indicated that the high level of pH3T6, pH3S10 and pH3Y41 are signatures associated with a poor prognosis of overall survival (OS) of GBM treated with the "temozolomide and irradiation standard" treatment of GBM (named TMZ+Irad treatment). Our data also showed that these signatures are correlated with the high activity of kinases already described as writers of the pH3T6, pH3S10 and pH3Y41 i.e. the PKC, Aurora-B and JAK2, respectively. Finally, our analysis revealed that the use of Enzastaurin, AZD1152, and AZD1480 abrogated the high level of pH3T6, pH3S10 and pH3Y41 while increasing the sensitivity to the “temozolomide and irradiation”-induced cell death. To conclude, it appears that this work provides biomarkers for patient stratification for a therapy including kinase inhibitors. PMID:25553095

  15. Influence of forage harvesting regimes on dynamics of biological dinitrogen fixation of a tropical woody legume.

    PubMed

    Nygren, Pekka; Cruz, Pablo; Domenach, Anne Marie; Vaillant, Victor; Sierra, Jorge

    2000-01-01

    Effects of three forage harvesting regimes-total removal of foliage and branches once (T-12) or twice a year (T-6) and 50% removal every 2 months (P-2)-on growth and biological dinitrogen fixation of Gliricidia sepium (Jacq.) Walp were studied under subhumid tropical conditions in Guadeloupe, French Antilles. Gliricidia sepium was grown in association with the perennial C(4) grass Dichantium aristatum (Poir) C.E. Hubbard in a two-storied fodder production system. The medium-term effects of pruning on N(2) fixation were assessed by the (15)N natural abundance method. Gmelina arborea Roxb. was used as the non-fixing reference. The trees in the T-12 regime followed the natural phenological cycle, and flowering and podfilling at the beginning of the dry season reduced both foliage and nodule biomass. The T-6 regime impeded flowering, and only a few flowers, on older branches, were produced in the P-2 regime. In trees in the T-12, T-6, and P-2 regimes, fixed N comprised 54-87, 54-92, and 60-87%, respectively, of the total N in aboveground biomass, depending on sampling date. Total annual accumulation of N in harvestable aboveground biomass was highest in trees in the T-6 regime at 313 kg ha(-1), of which 204 kg ha(-1) of N was fixed from the atmosphere. In all treatments, about 70% of the N exported per year from the plot in the fodder harvest came from N(2) fixation. Thus, N(2) fixation makes an important contribution to the N economy of the G. sepium-D. aristatum forage production system, and greatly reduces the need for fertilizer application. PMID:12651525

  16. Corrosion Behaviour of A380 Aluminiun Alloy by Semi-Solid Rheocasting

    SciTech Connect

    Forn, A.; Ruperez, E.; Baile, M. T.; Campillo, M.; Menargues, S.; Espinosa, I.

    2007-04-07

    A comparative study was performed on the corrosion behavior of a component of A380 aluminium alloy obtained by Semi-Solid Rheocasting (SSR). The effect of heat treatments T5 and T6 on corrosion resistance was compared with components without heat treatment by SSR processes. Corrosion studies were performed using an acetic acid salt spray test, impedance measurements and polarization curves using a 3,5%Na Cl test solution. The corrosion progress is described by micrographic analysis.

  17. Defective Initiation of Glycosaminoglycan Synthesis due to B3GALT6 Mutations Causes a Pleiotropic Ehlers-Danlos-Syndrome-like Connective Tissue Disorder

    PubMed Central

    Malfait, Fransiska; Kariminejad, Ariana; Van Damme, Tim; Gauche, Caroline; Syx, Delfien; Merhi-Soussi, Faten; Gulberti, Sandrine; Symoens, Sofie; Vanhauwaert, Suzanne; Willaert, Andy; Bozorgmehr, Bita; Kariminejad, Mohamad Hasan; Ebrahimiadib, Nazanin; Hausser, Ingrid; Huysseune, Ann; Fournel-Gigleux, Sylvie; De Paepe, Anne

    2013-01-01

    Proteoglycans are important components of cell plasma membranes and extracellular matrices of connective tissues. They consist of glycosaminoglycan chains attached to a core protein via a tetrasaccharide linkage, whereby the addition of the third residue is catalyzed by galactosyltransferase II (β3GalT6), encoded by B3GALT6. Homozygosity mapping and candidate gene sequence analysis in three independent families, presenting a severe autosomal-recessive connective tissue disorder characterized by skin fragility, delayed wound healing, joint hyperlaxity and contractures, muscle hypotonia, intellectual disability, and a spondyloepimetaphyseal dysplasia with bone fragility and severe kyphoscoliosis, identified biallelic B3GALT6 mutations, including homozygous missense mutations in family 1 (c.619G>C [p.Asp207His]) and family 3 (c.649G>A [p.Gly217Ser]) and compound heterozygous mutations in family 2 (c.323_344del [p.Ala108Glyfs∗163], c.619G>C [p.Asp207His]). The phenotype overlaps with several recessive Ehlers-Danlos variants and spondyloepimetaphyseal dysplasia with joint hyperlaxity. Affected individuals’ fibroblasts exhibited a large decrease in ability to prime glycosaminoglycan synthesis together with impaired glycanation of the small chondroitin/dermatan sulfate proteoglycan decorin, confirming β3GalT6 loss of function. Dermal electron microcopy disclosed abnormalities in collagen fibril organization, in line with the important regulatory role of decorin in this process. A strong reduction in heparan sulfate level was also observed, indicating that β3GalT6 deficiency alters synthesis of both main types of glycosaminoglycans. In vitro wound healing assay revealed a significant delay in fibroblasts from two index individuals, pointing to a role for glycosaminoglycan defect in impaired wound repair in vivo. Our study emphasizes a crucial role for β3GalT6 in multiple major developmental and pathophysiological processes. PMID:23664118

  18. [Effects of nitrogen management on yield, quality, nitrogen accumulation and its transportation of watermelon in gravel-mulched field].

    PubMed

    Ma, Zhong-ming; Du, Shao-ping; Xue, Liang

    2015-11-01

    The effects of nitrogen management on yield, quality, nitrogen and dry matter accumulation and transportation of watermelon in sand field were studied based on a field experiment. The results showed that too low or too high basal nitrogen fertilzation was unfavorable to seedling growth of watermelon in sand field, and no nitrogen application at vine extension or fruiting stages limited the formation of 'source' or 'sink'. At the same nitrogen rate, compared with the traditional T1 treatment (30% basal N fertilizer + 70% N fertilizer in vine extension), the nitrogen and dry matter accumulation of vegetative organs of T4 treatment (30% basal N fertilizer + 30% N fertilizer in vine extension + 40% N fertilizer in fruiting) and T6 treatment (100% basal N fertilizer + NAM) were reduced significantly, but the nitrogen and dry matter accumulation of fruit were increased significantly in the flushing period. The nitrogen transportation ratio and nitrogen contribution ratio of T4 were 33.6% and 12.0%, respectively. Compared to T1, the nitrogen harvest index, nitrogen fertilizer partial factor productivity and nitrogen fertilizer recovery efficiency of T4 and T6 treatments increased by 14.1% and 12.7%, 11.6% and 12.5%, 5.3% and 8.7%, respectively, and yield of watermelon increased by 11.6% and 12.5%, the soluble sugar, effective acid, the ratio of sugar and acid, Vc content increased by 16.5% and 11.7%, 4.5% and 2.8%, 19.4% and 13.4%, 35.6% and 19.0%, respectively. Therefore, T4 and T6 treatments were the optimal nitrogen fertilizer management mode which could not only achieve high yield and quality but also obtain high nitrogen fertilizer use efficiency in sand field. T6 treatment was the best nitrogen fertilizer management mode considering reduction of fertilizing labor intensity and extending service time of gravel-mulched field. PMID:26915190

  19. Vibrio cholerae Response Regulator VxrB Controls Colonization and Regulates the Type VI Secretion System.

    PubMed

    Cheng, Andrew T; Ottemann, Karen M; Yildiz, Fitnat H

    2015-05-01

    Two-component signal transduction systems (TCS) are used by bacteria to sense and respond to their environment. TCS are typically composed of a sensor histidine kinase (HK) and a response regulator (RR). The Vibrio cholerae genome encodes 52 RR, but the role of these RRs in V. cholerae pathogenesis is largely unknown. To identify RRs that control V. cholerae colonization, in-frame deletions of each RR were generated and the resulting mutants analyzed using an infant mouse intestine colonization assay. We found that 12 of the 52 RR were involved in intestinal colonization. Mutants lacking one previously uncharacterized RR, VCA0566 (renamed VxrB), displayed a significant colonization defect. Further experiments showed that VxrB phosphorylation state on the predicted conserved aspartate contributes to intestine colonization. The VxrB regulon was determined using whole genome expression analysis. It consists of several genes, including those genes that create the type VI secretion system (T6SS). We determined that VxrB is required for T6SS expression using several in vitro assays and bacterial killing assays, and furthermore that the T6SS is required for intestinal colonization. vxrB is encoded in a four gene operon and the other vxr operon members also modulate intestinal colonization. Lastly, though ΔvxrB exhibited a defect in single-strain intestinal colonization, the ΔvxrB strain did not show any in vitro growth defect. Overall, our work revealed that a small set of RRs is required for intestinal colonization and one of these regulators, VxrB affects colonization at least in part through its regulation of T6SS genes.

  20. A training course on food hygiene for butchers: measuring its effectiveness through microbiological analysis and the use of an inspection checklist.

    PubMed

    Vaz, Maria Luiza Santomauro; Novo, Neil Ferreira; Sigulem, Dirce Maria; Morais, Tania Beninga

    2005-11-01

    The effectiveness of food hygiene training for a group of retail butchers was evaluated with the aim of verifying whether the butchers modified their behavior in the light of knowledge gained and whether their acquired knowledge or behavior change was sustained over a period of time. Microbiological analysis (enumeration of mesophilic and coliform bacteria and Escherichia coli) of a raw semiprocessed product (stuffed rolled beef) was conducted, and an inspection checklist was issued before the training course (T0). Initial results were later compared with results obtained 1 month (T1) and 6 months (T6) after the training. The checklist comprised 89 items classified into five categories: A, approved suppliers and product reception; B, storage conditions and temperature control; C, flow process, food handling procedures, and conditions of the window display unit; D, facility design and proper cleaning and sanitizing of equipment, utensils, and work surfaces; and E, pest control system, water supply control, and garbage disposal. The inspection results were recorded as "yes" or "no" for each item. Compliance with food safety procedures was recorded as the percentage of "yes" answers. The bacterial counts were significantly higher at T0. At T6, there was no significant increase in bacterial counts. There was a significant improvement in food safety practices at T1 and T6 compared with T0 for all categories. When comparing T0 and T1, the largest increases in the compliance scores were seen within categories C and D. No significant decrease in scores for compliance with food safety practices was observed at T6. Supervision and refresher activities may be necessary to maintain behavioral changes for a longer period of time.

  1. Screening for Inhibition of Vibrio cholerae VipA-VipB Interaction Identifies Small-Molecule Compounds Active against Type VI Secretion

    PubMed Central

    Sun, Kun; Bröms, Jeanette; Lavander, Moa; Gurram, Bharat Kumar; Enquist, Per-Anders; Andersson, C. David; Elofsson, Mikael

    2014-01-01

    The type VI secretion system (T6SS) is the most prevalent bacterial secretion system and an important virulence mechanism utilized by Gram-negative bacteria, either to target eukaryotic cells or to combat other microbes. The components show much variability, but some appear essential for the function, and two homologues, denoted VipA and VipB in Vibrio cholerae, have been identified in all T6SSs described so far. Secretion is dependent on binding of an α-helical region of VipA to VipB, and in the absence of this binding, both components are degraded within minutes and secretion is ceased. The aim of the study was to investigate if this interaction could be blocked, and we hypothesized that such inhibition would lead to abrogation of T6S. A library of 9,600 small-molecule compounds was screened for their ability to block the binding of VipA-VipB in a bacterial two-hybrid system (B2H). After excluding compounds that showed cytotoxicity toward eukaryotic cells, that inhibited growth of Vibrio, or that inhibited an unrelated B2H interaction, 34 compounds were further investigated for effects on the T6SS-dependent secretion of hemolysin-coregulated protein (Hcp) or of phospholipase A1 activity. Two compounds, KS100 and KS200, showed intermediate or strong effects in both assays. Analogues were obtained, and compounds with potent inhibitory effects in the assays and desirable physicochemical properties as predicted by in silico analysis were identified. Since the compounds specifically target a virulence mechanism without affecting bacterial replication, they have the potential to mitigate the virulence with minimal risk for development of resistance. PMID:24798289

  2. Dye penetrant indications caused by superficial surface defects in 2014 aluminum alloy welds.

    NASA Technical Reports Server (NTRS)

    Hocker, R. G.; Wilson, K. R.

    1971-01-01

    Demonstration that dye penetrant indications on the heat-affected zone of 2014-T6 aluminum GMA weldments are frequently caused by superficial surface conditions and are less than 0.007 in. deep. The following methods are suggested for minimization of these surface defects: stabilization of the arc, application of dc ?GTA' welding procedures, reduction of the caustic etch time, and use of fine grain materials.

  3. Structural model for gamma-aminobutyric acid receptor noncompetitive antagonist binding: widely diverse structures fit the same site.

    PubMed

    Chen, Ligong; Durkin, Kathleen A; Casida, John E

    2006-03-28

    Several major insecticides, including alpha-endosulfan, lindane, and fipronil, and the botanical picrotoxinin are noncompetitive antagonists (NCAs) for the GABA receptor. We showed earlier that human beta(3) homopentameric GABA(A) receptor recognizes all of the important GABAergic insecticides and reproduces the high insecticide sensitivity and structure-activity relationships of the native insect receptor. Despite large structural diversity, the NCAs are proposed to fit a single binding site in the chloride channel lumen lined by five transmembrane 2 segments. This hypothesis is examined with the beta(3) homopentamer by mutagenesis, pore structure studies, NCA binding, and molecular modeling. The 15 amino acids in the cytoplasmic half of the pore were mutated to cysteine, serine, or other residue for 22 mutants overall. Localization of A-1'C, A2'C, T6'C, and L9'C (index numbers for the transmembrane 2 region) in the channel lumen was established by disulfide cross-linking. Binding of two NCA radioligands [(3)H]1-(4-ethynylphenyl)-4-n-propyl-2,6,7-trioxabicyclo[2.2.2]octane and [(3)H] 3,3-bis-trifluoromethyl-bicyclo[2,2,1]heptane-2,2-dicarbonitrile was dramatically reduced with 8 of the 15 mutated positions, focusing attention on A2', T6', and L9' as proposed binding sites, consistent with earlier mutagenesis studies. The cytoplasmic half of the beta3 homopentamer pore was modeled as an alpha-helix. The six NCAs listed above plus t-butylbicyclophosphorothionate fit the 2' to 9' pore region forming hydrogen bonds with the T6' hydroxyl and hydrophobic interactions with A2', T6', and L9' alkyl substituents, thereby blocking the channel. Thus, widely diverse NCA structures fit the same GABA receptor beta subunit site with important implications for insecticide cross-resistance and selective toxicity between insects and mammals.

  4. Temperature field acquisition during gas metal arc welding using thermocouples, thermography and fibre Bragg grating sensors

    NASA Astrophysics Data System (ADS)

    Moreira, P. M. G. P.; Frazão, O.; Tavares, S. M. O.; de Figueiredo, M. A. V.; Restivo, M. T.; Santos, J. L.; de Castro, P. M. S. T.

    2007-03-01

    The paper presents the application of temperature acquisition systems integrating thermocouples, a thermographic camera and fibre Bragg grating (FBG) sensors in gas metal arc welding (GMAW) process, MIG (metal inert gas) welding type. Efficient procedures to use FBG sensors and thermocouples were developed. The paper presents and compares measurements made in welded plates of aluminium alloy 6082-T6. Tests were performed in both plate surfaces and good agreement between the three techniques was found.

  5. Initial Subdivision of Genesis Early Science Polished Aluminum Collector

    NASA Technical Reports Server (NTRS)

    Allton, J. H.; Stansbery, E. K.; McNamara, K. M.; Meshik, A.; See, T. H.; Bastien, R.

    2005-01-01

    A large surface, about 245 square centimeters, of highly polished aluminum 6061 T6 alloy was attached to the science canister thermal panel for the purpose of collecting solar wind noble gases. The analysis of this collector will be part of the Genesis Early Science results. The pre-launch configuration of the collector is shown. The collector sustained some damage during the recovery impact in Utah, September 8, 2004.

  6. Paraplegia after thoracotomy for division and suture Patent Ductus Arteriosus (PDA).

    PubMed

    Sayasathid, Jarun; Somboonna, Naraporn; Numchaisiri, Chun

    2006-12-01

    A Thai women, aged 22 years old, came to hospital with Patent Ductus Arteriosis (PDA). Left thoracotomy, with division and suturing PDA, was performed. The second day after operation, she developed paraplegia below umbilical level. The CT-scan detected an extradural hematoma in the spinal cavity from T3-T6. To remove the blood clot, the T spine laminectomy was performed. 6 months after the laminectomy, the patient was able to perform her regular exercise. PMID:17214069

  7. Cyclic Deformation Behavior of a Rare-Earth Containing Extruded Magnesium Alloy: Effect of Heat Treatment

    NASA Astrophysics Data System (ADS)

    Mirza, F. A.; Chen, D. L.; Li, D. J.; Zeng, X. Q.

    2015-03-01

    The present study was aimed at evaluating strain-controlled cyclic deformation behavior of a rare-earth (RE) element containing Mg-10Gd-3Y-0.5Zr (GW103K) alloy in different states (as-extruded, peak-aged (T5), and solution-treated and peak-aged (T6)). The addition of RE elements led to an effective grain refinement and weak texture in the as-extruded alloy. While heat treatment resulted in a grain growth modestly in the T5 state and significantly in the T6 state, a high density of nano-sized and bamboo-leaf/plate-shaped β' (Mg7(Gd,Y)) precipitates was observed to distribute uniformly in the α-Mg matrix. The yield strength and ultimate tensile strength, as well as the maximum and minimum peak stresses during cyclic deformation in the T5 and T6 states were significantly higher than those in the as-extruded state. Unlike RE-free extruded Mg alloys, symmetrical hysteresis loops in tension and compression and cyclic stabilization were present in the GW103K alloy in different states. The fatigue life of this alloy in the three conditions, which could be well described by the Coffin-Manson law and Basquin's equation, was equivalent within the experimental scatter and was longer than that of RE-free extruded Mg alloys. This was predominantly attributed to the presence of the relatively weak texture and the suppression of twinning activities stemming from the fine grain sizes and especially RE-containing β' precipitates. Fatigue crack was observed to initiate from the specimen surface in all the three alloy states and the initiation site contained some cleavage-like facets after T6 heat treatment. Crack propagation was characterized mainly by the characteristic fatigue striations.

  8. Drug membrane transporters and CYP3A4 are affected by hypericin, hyperforin or aristoforin in colon adenocarcinoma cells.

    PubMed

    Šemeláková, M; Jendželovský, R; Fedoročko, P

    2016-07-01

    Our previous results have shown that the combination of hypericin-mediated photodynamic therapy (HY-PDT) at sub-optimal dose with hyperforin (HP) (compounds of Hypericum sp.), or its stable derivative aristoforin (AR) stimulates generation of reactive oxygen species (ROS) leading to antitumour activity. This enhanced oxidative stress evoked the need for an explanation for HY accumulation in colon cancer cells pretreated with HP or AR. Generally, the therapeutic efficacy of chemotherapeutics is limited by drug resistance related to the overexpression of drug efflux transporters in tumour cells. Therefore, the impact of non-activated hypericin (HY), HY-PDT, HP and AR on cell membrane transporter systems (Multidrug resistance-associated protein 1-MRP1/ABCC1, Multidrug resistance-associated protein 2-MRP2/ABCC2, Breast cancer resistance protein - BCRP/ABCG2, P-glycoprotein-P-gp/ABCC1) and cytochrome P450 3A4 (CYP3A4) was evaluated. The different effects of the three compounds on their expression, protein level and activity was determined under specific PDT light (T0+, T6+) or dark conditions (T0- T6-). We found that HP or AR treatment affected the protein levels of MRP2 and P-gp, whereas HP decreased MRP2 and P-gp expression mostly in the T0+ and T6+ conditions, while AR decreased MRP2 in T0- and T6+. Moreover, HY-PDT treatment induced the expression of MRP1. Our data demonstrate that HP or AR treatment in light or dark PDT conditions had an inhibitory effect on the activity of individual membrane transport proteins and significantly decreased CYP3A4 activity in HT-29 cells. We found that HP or AR significantly affected intracellular accumulation of HY in HT-29 colon adenocarcinoma cells. These results suggest that HY, HP and AR might affect the efficiency of anti-cancer drugs, through interaction with membrane transporters and CYP3A4. PMID:27261575

  9. An invasive and low virulent Edwardsiella tarda esrB mutant promising as live attenuated vaccine in aquaculture.

    PubMed

    Yang, Weizheng; Wang, Lixia; Zhang, Lingzhi; Qu, Jiangbo; Wang, Qiyao; Zhang, Yuanxing

    2015-02-01

    Edwardsiella tarda is a leading fish pathogen haunting worldwide aquaculture industry. In E. tarda, two-component system EsrA-EsrB positively regulates type III and VI secretion systems (T3SS and T6SS) and negatively regulates hemolysin EthA, which has been demonstrated to be essential for the invasion processes in fish. In order to develop a live attenuated vaccine (LAV) with high invasiveness to be practically and economically used as immersion-administered vaccine in aquaculture, here, we generated a random mutation library of esrB sequences by error-prone PCR and introduced them into the E. tarda esrB deletion mutant. The mutant YWZ47 with significantly increased hemolytic activity and low T3SS and T6SS secretion was screened. Phenotypes including extracellular protein profiles, invasion in macrophages, lethality toward fish, and infection kinetics were investigated in the wild-type strain EIB202 and the mutants ΔesrB, ΔT3SS, ΔT6SS, ΔT3SS/ΔT6SS, and YWZ47. Compared to the documented LAV strain ΔesrB, YWZ47 showed higher invasive capability and low in vivo virulence toward fish. Significantly higher relative percent survival (RPS) could be generated in turbot (Scophthalmus maximus) against the challenge of the wild-type EIB202 when inoculated through immersion route, and the RPS was comparable with that of ΔesrB through intraperitoneal (i.p.) injection inoculation. Two mutated points, K167M and H197L, were found by sequence analysis of EsrBYWZ47 variant. These structural modifications underpin the variations in the regulatory functions of the mutant and wild-type EsrB. This study promoted understanding of virulence regulation by EsrB in E. tarda and presented a promising candidate of invasive attenuated vaccine used in aquaculture industries.

  10. Evaluation of Chang's culture medium for mouse in vitro fertilization and embryonic development.

    PubMed

    Ariff, B; Ng, S C; Mok, H; Lim, M N; Wong, P C; Shan, R

    1988-04-01

    Chang's medium [with and without human serum (HS)] was compared with T6 medium [with and without bovine serum albumin (BSA)] for in vitro fertilization (IVF) and development of two-cell mouse embryos to the blastocyst stage. Chang's medium without any supplementation gave significantly better fertilization rates (83.3%) than Chang's with 10% HS (76.4%) or T6 and BSA (76.6%) (P less than 0.01). In a separate experiment 87.7% of the two-cell mouse embryos developed to the blastocyst stage in Chang's medium, compared to 90.6% for T6 with BSA and 93.6% without BSA, respectively (P greater than 0.01). In Chang's medium supplemented with 10% HS, 76.6% of the embryos developed to the blastocyst stage and 17.2% stopped development after the morula stage. After 72 hr in vitro hatched trophoblast and inner-cell-mass cells from 26.5 and 30.8% of the embryos grown in Chang's medium (with and without HS) attached to the plastic culture dishes and grew to form a mixed monolayer of epithelioid and fibroblastic cells. Chang's medium can thus be successfully used for IVF and growth of mammalian embryos. Further, inner cell mass and trophoblast cell lines could be established for various reproductive studies using this medium.

  11. Comparative Genomics of Multiple Strains of Pseudomonas cannabina pv. alisalensis, a Potential Model Pathogen of Both Monocots and Dicots

    PubMed Central

    Sarris, Panagiotis F.; Trantas, Emmanouil A.; Baltrus, David A.; Bull, Carolee T.; Wechter, William Patrick; Yan, Shuangchun; Ververidis, Filippos; Almeida, Nalvo F.; Jones, Corbin D.; Dangl, Jeffery L.; Panopoulos, Nickolas J.; Vinatzer, Boris A.; Goumas, Dimitrios E.

    2013-01-01

    Comparative genomics of closely related pathogens that differ in host range can provide insights into mechanisms of host-pathogen interactions and host adaptation. Furthermore, sequencing of multiple strains with the same host range reveals information concerning pathogen diversity and the molecular basis of virulence. Here we present a comparative analysis of draft genome sequences for four strains of Pseudomonas cannabina pathovar alisalensis (Pcal), which is pathogenic on a range of monocotyledonous and dicotyledonous plants. These draft genome sequences provide a foundation for understanding host range evolution across the monocot-dicot divide. Like other phytopathogenic pseudomonads, Pcal strains harboured a hrp/hrc gene cluster that codes for a type III secretion system. Phylogenetic analysis based on the hrp/hrc cluster genes/proteins, suggests localized recombination and functional divergence within the hrp/hrc cluster. Despite significant conservation of overall genetic content across Pcal genomes, comparison of type III effector repertoires reinforced previous molecular data suggesting the existence of two distinct lineages within this pathovar. Furthermore, all Pcal strains analyzed harbored two distinct genomic islands predicted to code for type VI secretion systems (T6SSs). While one of these systems was orthologous to known P. syringae T6SSs, the other more closely resembled a T6SS found within P. aeruginosa. In summary, our study provides a foundation to unravel Pcal adaptation to both monocot and dicot hosts and provides genetic insights into the mechanisms underlying pathogenicity. PMID:23555661

  12. Preliminary fracture analysis of the core pressure boundary tube for the Advanced Neutron Source Research Reactor

    SciTech Connect

    Schulz, K.C.; Yahr, G.T.

    1995-08-01

    The outer core pressure boundary tube (CPBT) of the Advanced neutron Source (ANS) reactor being designed at Oak Ridge National Laboratory is currently specified as being composed of 6061-T6 aluminum. ASME Boiler and Pressure Vessel Code fracture analysis rules for nuclear components are based on the use of ferritic steels; the expressions, tables, charts and equations were all developed from tests and analyses conducted for ferritic steels. Because of the nature of the Code, design with thin aluminum requires analytical approaches that do not directly follow the Code. The intent of this report is to present a methodology comparable to the ASME Code for ensuring the prevention of nonductile fracture of the CPBT in the ANS reactor. 6061-T6 aluminum is known to be a relatively brittle material; the linear elastic fracture mechanics (LEFM) approach is utilized to determine allowable flaw sizes for the CPBT. A J-analysis following the procedure developed by the Electric Power Research Institute was conducted as a check; the results matched those for the LEFM analysis for the cases analyzed. Since 6061-T6 is known to embrittle when irradiated, the reduction in K{sub Q} due to irradiation is considered in the analysis. In anticipation of probable requirements regarding maximum allowable flaw size, a survey of nondestructive inspection capabilities is also presented. A discussion of probabilistic fracture mechanics approaches, principally Monte Carlo techniques, is included in this report as an introduction to what quantifying the probability of nonductile failure of the CPBT may entail.

  13. Morphologies of fibroblast cells cultured on surfaces of PHB films implanted by hydroxyl ions.

    PubMed

    Hou, T; Zhang, J Z; Kong, L J; Zhang, X E; Hu, P; Zhang, D M; Li, N

    2006-01-01

    Polyhydroxybutyrate (PHB) films were implanted with 40 keV hydroxyl ions with fluences ranging from 1 x 10(12) to 1 x 10(15) ions/cm2, respectively. The as-implanted PHB films were characterized by scanning electron microscopy (SEM), electron spectroscopy for chemical analysis (ESCA) and water contact angle measurements. The surface structures and properties of the as-implanted PHB films were closely related with hydroxyl ion fluence. They were further investigated by inoculating 3T6 fibroblasts cells on their surfaces. Morphologies of the 3T6 fibroblast cells cultured on surfaces of the as-implanted PHB films were observed by SEM. Characterization of the cultural 3T6 cells was analyzed qualitatively. The preliminary experimental results reveal that the bioactivity of the PHB films modified by hydroxyl ion implantation was improved at different levels, and the fluence of 1 x 10(13) ions/cm2 is optimal for PHB film. PMID:16909942

  14. Trehalose-6-phosphate synthesis controls yeast gluconeogenesis downstream and independent of SNF1.

    PubMed

    Deroover, Sofie; Ghillebert, Ruben; Broeckx, Tom; Winderickx, Joris; Rolland, Filip

    2016-06-01

    Trehalose-6-P (T6P), an intermediate of trehalose biosynthesis, was identified as an important regulator of yeast sugar metabolism and signaling. tps1Δ mutants, deficient in T6P synthesis (TPS), are unable to grow on rapidly fermentable medium with uncontrolled influx in glycolysis, depletion of ATP and accumulation of sugar phosphates. However, the exact molecular mechanisms involved are not fully understood. We show that SNF1 deletion restores the tps1Δ growth defect on glucose, suggesting that lack of TPS hampers inactivation of SNF1 or SNF1-regulated processes. In addition to alternative, non-fermentable carbon metabolism, SNF1 controls two major processes: respiration and gluconeogenesis. The tps1Δ defect appears to be specifically associated with deficient inhibition of gluconeogenesis, indicating more downstream effects. Consistently, Snf1 dephosphorylation and inactivation on glucose medium are not affected, as confirmed with an in vivo Snf1 activity reporter. Detailed analysis shows that gluconeogenic Pck1 and Fbp1 expression, protein levels and activity are not repressed upon glucose addition to tps1Δ cells, suggesting a link between the metabolic defect and persistent gluconeogenesis. While SNF1 is essential for induction of gluconeogenesis, T6P/TPS is required for inactivation of gluconeogenesis in the presence of glucose, downstream and independent of SNF1 activity and the Cat8 and Sip4 transcription factors.

  15. Crystal Structure of Hcp from Acinetobacter baumannii: A Component of the Type VI Secretion System

    PubMed Central

    Ruiz, Federico M.; Santillana, Elena; Spínola-Amilibia, Mercedes; Torreira, Eva; Culebras, Esther; Romero, Antonio

    2015-01-01

    The type VI secretion system (T6SS) is a bacterial macromolecular machine widely distributed in Gram-negative bacteria, which transports effector proteins into eukaryotic host cells or other bacteria. Membrane complexes and a central tubular structure, which resembles the tail of contractile bacteriophages, compose the T6SS. One of the proteins forming this tube is the hemolysin co-regulated protein (Hcp), which acts as virulence factor, as transporter of effectors and as a chaperone. In this study, we present the structure of Hcp from Acinetobacter baumannii, together with functional and oligomerization studies. The structure of this protein exhibits a tight β barrel formed by two β sheets and flanked at one side by a short α-helix. Six Hcp molecules associate to form a donut-shaped hexamer, as observed in both the crystal structure and solution. These results emphasize the importance of this oligomerization state in this family of proteins, despite the low similarity of sequence among them. The structure presented in this study is the first one for a protein forming part of a functional T6SS from A. baumannii. These results will help us to understand the mechanism and function of this secretion system in this opportunistic nosocomial pathogen. PMID:26079269

  16. The translocation (6;9), associated with a specific subtype of acute myeloid leukemia, results in the fusion of two genes, dek and can, and the expression of a chimeric, leukemia-specific dek-can mRNA.

    PubMed Central

    von Lindern, M; Fornerod, M; van Baal, S; Jaegle, M; de Wit, T; Buijs, A; Grosveld, G

    1992-01-01

    The translocation (6;9) is associated with a specific subtype of acute myeloid leukemia (AML). Previously, it was found that breakpoints on chromosome 9 are clustered in one of the introns of a large gene named Cain (can). cDNA probes derived from the 3' part of can detect an aberrant, leukemia-specific 5.5-kb transcript in bone marrow cells from t(6;9) AML patients. cDNA cloning of this mRNA revealed that it is a fusion of sequences encoded on chromosome 6 and 3' can. A novel gene on chromosome 6 which was named dek was isolated. In dek the t(6;9) breakpoints also occur in one intron. As a result the dek-can fusion gene, present in t(6;9) AML, encodes an invariable dek-can transcript. Sequence analysis of the dek-can cDNA showed that dek and can are merged without disruption of the original open reading frames and therefore the fusion mRNA encodes a chimeric DEK-CAN protein of 165 kDa. The predicted DEK and CAN proteins have molecular masses of 43 and 220 kDa, respectively. Sequence comparison with the EMBL data base failed to show consistent homology with any known protein sequences. Images PMID:1549122

  17. T(lys), a newly identified Sulfolobus spindle-shaped virus 1 transcript expressed in the lysogenic state, encodes a DNA-binding protein interacting at the promoters of the early genes.

    PubMed

    Fusco, Salvatore; She, Qunxin; Bartolucci, Simonetta; Contursi, Patrizia

    2013-05-01

    While studying the gene expression of the Sulfolobus spindle-shaped virus 1 (SSV1) in Sulfolobus solfataricus lysogenic cells, a novel viral transcript (T(lys)) was identified. Transcriptional analysis revealed that T(lys) is expressed only in the absence of UV irradiation and is downregulated during the growth of the lysogenic host. The correponding gene f55 lies between two transcriptional units (T6 and T(ind)) that are upregulated upon UV irradiation. The open reading frame f55 encodes a 6.3-kDa protein which shows sequence identity with negative regulators that fold into the ribbon-helix-helix DNA-binding motif. DNA-binding assays demonstrated that the recombinant F55, purified from Escherichia coli, is indeed a putative transcription factor able to recognize site specifically target sequences in the promoters of the early induced T5, T6, and T(ind) transcripts, as well as of its own promoter. Binding sites of F55 are included within a tandem-repeated sequence overlapping the transcription start sites and/or the B recognition element of the pertinent genes. The strongest binding was observed with the promoters of T5 and T6, and an apparent cooperativity in binding was observed with the T(ind) promoter. Taking together the transcriptional analysis data and the biochemical evidences, we surmise that the protein F55 is involved in the regulation of the lysogenic state of SSV1. PMID:23514883

  18. Hormone therapy with tamoxifen reduces plasma levels of NT-B-type natriuretic peptide but does not change ventricular ejection fraction after chemotherapy in women with breast cancer.

    PubMed

    Silva, F B; Romero, W G; Carvalho, A L R; Borgo, M V; Amorim, M H C; Gouvea, S A; Abreu, G R

    2015-02-01

    The objective of this study was to evaluate the effect of tamoxifen on the plasma concentration of NT-pro-B-type natriuretic peptide (NT-proBNP) in women undergoing chemotherapy for breast cancer and to correlate changes in NT-proBNP with the left ventricular ejection fraction (LVEF). Over a period of 12 months, we followed 60 women with a diagnosis of breast cancer. The patients were separated into a group that received only chemotherapy (n=23), a group that received chemotherapy + tamoxifen (n=21), and a group that received only tamoxifen (n=16). Plasma levels of NT-proBNP were assessed at 0 (T0), 6 (T6), and 12 (T12) months of treatment, and echocardiography data were assessed at T0 and T12. Plasma NT-proBNP levels were increased in the chemotherapy-only group at T6 and T12, whereas elevated NT-proBNP levels were only found at T6 in the chemotherapy + tamoxifen group. At T12, the chemotherapy + tamoxifen group exhibited a significant reduction in the peptide to levels similar to the group that received tamoxifen alone. The chemotherapy-only group exhibited a significant decrease in LVEF at T12, whereas the chemotherapy + tamoxifen and tamoxifen-only groups maintained levels similar to those at the beginning of treatment. Treatment with tamoxifen for 6 months after chemotherapy significantly reduced the plasma levels of NT-proBNP and did not change LVEF in women with breast cancer.

  19. Identification of protein secretion systems in bacterial genomes

    PubMed Central

    Abby, Sophie S.; Cury, Jean; Guglielmini, Julien; Néron, Bertrand; Touchon, Marie; Rocha, Eduardo P. C.

    2016-01-01

    Bacteria with two cell membranes (diderms) have evolved complex systems for protein secretion. These systems were extensively studied in some model bacteria, but the characterisation of their diversity has lagged behind due to lack of standard annotation tools. We built online and standalone computational tools to accurately predict protein secretion systems and related appendages in bacteria with LPS-containing outer membranes. They consist of models describing the systems’ components and genetic organization to be used with MacSyFinder to search for T1SS-T6SS, T9SS, flagella, Type IV pili and Tad pili. We identified ~10,000 candidate systems in bacterial genomes, where T1SS and T5SS were by far the most abundant and widespread. All these data are made available in a public database. The recently described T6SSiii and T9SS were restricted to Bacteroidetes, and T6SSii to Francisella. The T2SS, T3SS, and T4SS were frequently encoded in single-copy in one locus, whereas most T1SS were encoded in two loci. The secretion systems of diderm Firmicutes were similar to those found in other diderms. Novel systems may remain to be discovered, since some clades of environmental bacteria lacked all known protein secretion systems. Our models can be fully customized, which should facilitate the identification of novel systems. PMID:26979785

  20. Whole genome sequencing provides insights into the genetic determinants of invasiveness in Salmonella Dublin.

    PubMed

    Mohammed, M; Cormican, M

    2016-08-01

    Salmonella enterica subsp. enterica serovar Dublin (S. Dublin) is one of the non-typhoidal Salmonella (NTS); however, a relatively high proportion of human infections are associated with invasive disease. We applied whole genome sequencing to representative invasive and non-invasive clinical isolates of S. Dublin to determine the genomic variations among them and to investigate the underlying genetic determinants associated with invasiveness in S. Dublin. Although no particular genomic variation was found to differentiate in invasive and non-invasive isolates four virulence factors were detected within the genome of all isolates including two different type VI secretion systems (T6SS) encoded on two Salmonella pathogenicity islands (SPI), including SPI-6 (T6SSSPI-6) and SPI-19 (T6SSSPI-19), an intact lambdoid prophage (Gifsy-2-like prophage) that contributes significantly to the virulence and pathogenesis of Salmonella serotypes in addition to a virulence plasmid. These four virulence factors may all contribute to the potential of S. Dublin to cause invasive disease in humans.

  1. Genetic Architecture of Abdominal Pigmentation in Drosophila melanogaster.

    PubMed

    Dembeck, Lauren M; Huang, Wen; Magwire, Michael M; Lawrence, Faye; Lyman, Richard F; Mackay, Trudy F C

    2015-05-01

    Pigmentation varies within and between species and is often adaptive. The amount of pigmentation on the abdomen of Drosophila melanogaster is a relatively simple morphological trait, which serves as a model for mapping the genetic basis of variation in complex phenotypes. Here, we assessed natural variation in female abdominal pigmentation in 175 sequenced inbred lines of the Drosophila melanogaster Genetic Reference Panel, derived from the Raleigh, NC population. We quantified the proportion of melanization on the two most posterior abdominal segments, tergites 5 and 6 (T5, T6). We found significant genetic variation in the proportion of melanization and high broad-sense heritabilities for each tergite. Genome-wide association studies identified over 150 DNA variants associated with the proportion of melanization on T5 (84), T6 (34), and the difference between T5 and T6 (35). Several of the top variants associated with variation in pigmentation are in tan, ebony, and bric-a-brac1, genes known to affect D. melanogaster abdominal pigmentation. Mutational analyses and targeted RNAi-knockdown showed that 17 out of 28 (61%) novel candidate genes implicated by the genome-wide association study affected abdominal pigmentation. Several of these genes are involved in developmental and regulatory pathways, chitin production, cuticle structure, and vesicle formation and transport. These findings show that genetic variation may affect multiple steps in pathways involved in tergite development and melanization. Variation in these novel candidates may serve as targets for adaptive evolution and sexual selection in D. melanogaster.

  2. Genetic Architecture of Abdominal Pigmentation in Drosophila melanogaster

    PubMed Central

    Dembeck, Lauren M.; Huang, Wen; Magwire, Michael M.; Lawrence, Faye; Lyman, Richard F.; Mackay, Trudy F. C.

    2015-01-01

    Pigmentation varies within and between species and is often adaptive. The amount of pigmentation on the abdomen of Drosophila melanogaster is a relatively simple morphological trait, which serves as a model for mapping the genetic basis of variation in complex phenotypes. Here, we assessed natural variation in female abdominal pigmentation in 175 sequenced inbred lines of the Drosophila melanogaster Genetic Reference Panel, derived from the Raleigh, NC population. We quantified the proportion of melanization on the two most posterior abdominal segments, tergites 5 and 6 (T5, T6). We found significant genetic variation in the proportion of melanization and high broad-sense heritabilities for each tergite. Genome-wide association studies identified over 150 DNA variants associated with the proportion of melanization on T5 (84), T6 (34), and the difference between T5 and T6 (35). Several of the top variants associated with variation in pigmentation are in tan, ebony, and bric-a-brac1, genes known to affect D. melanogaster abdominal pigmentation. Mutational analyses and targeted RNAi-knockdown showed that 17 out of 28 (61%) novel candidate genes implicated by the genome-wide association study affected abdominal pigmentation. Several of these genes are involved in developmental and regulatory pathways, chitin production, cuticle structure, and vesicle formation and transport. These findings show that genetic variation may affect multiple steps in pathways involved in tergite development and melanization. Variation in these novel candidates may serve as targets for adaptive evolution and sexual selection in D. melanogaster. PMID:25933381

  3. Tlys, a Newly Identified Sulfolobus Spindle-Shaped Virus 1 Transcript Expressed in the Lysogenic State, Encodes a DNA-Binding Protein Interacting at the Promoters of the Early Genes

    PubMed Central

    Fusco, Salvatore; She, Qunxin; Bartolucci, Simonetta

    2013-01-01

    While studying the gene expression of the Sulfolobus spindle-shaped virus 1 (SSV1) in Sulfolobus solfataricus lysogenic cells, a novel viral transcript (Tlys) was identified. Transcriptional analysis revealed that Tlys is expressed only in the absence of UV irradiation and is downregulated during the growth of the lysogenic host. The correponding gene f55 lies between two transcriptional units (T6 and Tind) that are upregulated upon UV irradiation. The open reading frame f55 encodes a 6.3-kDa protein which shows sequence identity with negative regulators that fold into the ribbon-helix-helix DNA-binding motif. DNA-binding assays demonstrated that the recombinant F55, purified from Escherichia coli, is indeed a putative transcription factor able to recognize site specifically target sequences in the promoters of the early induced T5, T6, and Tind transcripts, as well as of its own promoter. Binding sites of F55 are included within a tandem-repeated sequence overlapping the transcription start sites and/or the B recognition element of the pertinent genes. The strongest binding was observed with the promoters of T5 and T6, and an apparent cooperativity in binding was observed with the Tind promoter. Taking together the transcriptional analysis data and the biochemical evidences, we surmise that the protein F55 is involved in the regulation of the lysogenic state of SSV1. PMID:23514883

  4. Crystal Structure and Self-Interaction of the Type VI Secretion Tail-Tube Protein from Enteroaggregative Escherichia coli

    PubMed Central

    Douzi, Badreddine; Spinelli, Silvia; Blangy, Stéphanie; Roussel, Alain; Durand, Eric; Brunet, Yannick R.; Cascales, Eric; Cambillau, Christian

    2014-01-01

    The type VI secretion system (T6SS) is a widespread machine used by bacteria to control their environment and kill or disable bacterial species or eukaryotes through toxin injection. The T6SS comprises a central tube formed of stacked hexamers of hemolysin co-regulated proteins (Hcp) and terminated by a trimeric valine-glycine repeat protein G (VgrG) component, the cell puncturing device. A contractile tail sheath, formed by the TssB and TssC proteins, surrounds this tube. This syringe-like machine has been compared to an inverted phage, as both Hcp and VgrG share structural homology with tail components of Caudovirales. Here we solved the crystal structure of a tryptophan-substituted double mutant of Hcp1 from enteroaggregative Escherichia coli and compared it to the structures of other Hcps. Interestingly, we observed that the purified Hcp native protein is unable to form tubes in vitro. To better understand the rationale for observation, we measured the affinity of Hcp1 hexamers with themselves by surface plasmon resonance. The intra-hexamer interaction is weak, with a KD value of 7.2 µM. However, by engineering double cysteine mutants at defined positions, tubes of Hcp1 gathering up to 15 stacked hexamers formed in oxidative conditions. These results, together with those available in the literature regarding TssB and TssC, suggest that assembly of the T6SS tube differs significantly from that of Sipho- or Myoviridae. PMID:24551044

  5. Comparative Genome Analyses of Serratia marcescens FS14 Reveals Its High Antagonistic Potential

    PubMed Central

    Li, Pengpeng; Kwok, Amy H. Y.; Jiang, Jingwei; Ran, Tingting; Xu, Dongqing; Wang, Weiwu; Leung, Frederick C.

    2015-01-01

    S. marcescens FS14 was isolated from an Atractylodes macrocephala Koidz plant that was infected by Fusarium oxysporum and showed symptoms of root rot. With the completion of the genome sequence of FS14, the first comprehensive comparative-genomic analysis of the Serratia genus was performed. Pan-genome and COG analyses showed that the majority of the conserved core genes are involved in basic cellular functions, while genomic factors such as prophages contribute considerably to genome diversity. Additionally, a Type I restriction-modification system, a Type III secretion system and tellurium resistance genes are found in only some Serratia species. Comparative analysis further identified that S. marcescens FS14 possesses multiple mechanisms for antagonism against other microorganisms, including the production of prodigiosin, bacteriocins, and multi-antibiotic resistant determinants as well as chitinases. The presence of two evolutionarily distinct Type VI secretion systems (T6SSs) in FS14 may provide further competitive advantages for FS14 against other microbes. To our knowledge, this is the first report of comparative analysis on T6SSs in the genus, which identifies four types of T6SSs in Serratia spp.. Competition bioassays of FS14 against the vital plant pathogenic bacterium Ralstonia solanacearum and fungi Fusarium oxysporum and Sclerotinia sclerotiorum were performed to support our genomic analyses, in which FS14 demonstrated high antagonistic activities against both bacterial and fungal phytopathogens. PMID:25856195

  6. Comparative genomics of multiple strains of Pseudomonas cannabina pv. alisalensis, a potential model pathogen of both monocots and dicots.

    PubMed

    Sarris, Panagiotis F; Trantas, Emmanouil A; Baltrus, David A; Bull, Carolee T; Wechter, William Patrick; Yan, Shuangchun; Ververidis, Filippos; Almeida, Nalvo F; Jones, Corbin D; Dangl, Jeffery L; Panopoulos, Nickolas J; Vinatzer, Boris A; Goumas, Dimitrios E

    2013-01-01

    Comparative genomics of closely related pathogens that differ in host range can provide insights into mechanisms of host-pathogen interactions and host adaptation. Furthermore, sequencing of multiple strains with the same host range reveals information concerning pathogen diversity and the molecular basis of virulence. Here we present a comparative analysis of draft genome sequences for four strains of Pseudomonas cannabina pathovar alisalensis (Pcal), which is pathogenic on a range of monocotyledonous and dicotyledonous plants. These draft genome sequences provide a foundation for understanding host range evolution across the monocot-dicot divide. Like other phytopathogenic pseudomonads, Pcal strains harboured a hrp/hrc gene cluster that codes for a type III secretion system. Phylogenetic analysis based on the hrp/hrc cluster genes/proteins, suggests localized recombination and functional divergence within the hrp/hrc cluster. Despite significant conservation of overall genetic content across Pcal genomes, comparison of type III effector repertoires reinforced previous molecular data suggesting the existence of two distinct lineages within this pathovar. Furthermore, all Pcal strains analyzed harbored two distinct genomic islands predicted to code for type VI secretion systems (T6SSs). While one of these systems was orthologous to known P. syringae T6SSs, the other more closely resembled a T6SS found within P. aeruginosa. In summary, our study provides a foundation to unravel Pcal adaptation to both monocot and dicot hosts and provides genetic insights into the mechanisms underlying pathogenicity.

  7. Withdrawal of inhaled corticosteroids can be safe in COPD patients at low risk of exacerbation: a real-life study on the appropriateness of treatment in moderate COPD patients (OPTIMO)

    PubMed Central

    2014-01-01

    Background It has been suggested that withdrawal of inhaled corticosteroids (ICS) in COPD patients on maintenance treatment results in deterioration of symptoms, lung function and exacerbations. The aim of this real-life, prospective, multicentric study was to investigate whether withdrawal of ICS in COPD patients at low risk of exacerbation is linked to a deterioration in lung function and symptoms and to a higher frequency of exacerbations. Methods 914 COPD patients, on maintenance therapy with bronchodilators and ICS, FEV1>50% predicted, and <2 exacerbations/year were recruited. Upon decision of the primary physicians, 59% of patients continued their ICS treatment whereas in 41% of patients ICS were withdrawn and regular therapy was continued with long-acting bronchodilators mostly (91% of patients). FEV1, CAT (COPD Assessment Test), and occurrence of exacerbations were measured at the beginning (T0) and at the end (T6) of the 6 months observational period. Results 816 patients (89.3%) concluded the study. FEV1, CAT and exacerbations history were similar in the two groups (ICS and no ICS) at T0 and at T6. We did not observe any deterioration of lung function symptoms, and exacerbation rate between the two groups at T0 and T6. Conclusions We conclude that the withdrawal of ICS, in COPD patients at low risk of exacerbation, can be safe provided that patients are left on maintenance treatment with long-acting bronchodilators. PMID:25005873

  8. Involvement of type VI secretion system in secretion of iron chelator pyoverdine in Pseudomonas taiwanensis

    PubMed Central

    Chen, Wen-Jen; Kuo, Tzu-Yen; Hsieh, Feng-Chia; Chen, Pi-Yu; Wang, Chang-Sheng; Shih, Yu-Ling; Lai, Ying-Mi; Liu, Je-Ruei; Yang, Yu-Liang; Shih, Ming-Che

    2016-01-01

    Rice bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most destructive rice diseases worldwide. Therefore, in addition to breeding disease-resistant rice cultivars, it is desirable to develop effective biocontrol agents against Xoo. Here, we report that a soil bacterium Pseudomonas taiwanensis displayed strong antagonistic activity against Xoo. Using matrix-assisted laser desorption/ionization imaging mass spectrometry, we identified an iron chelator, pyoverdine, secreted by P. taiwanensis that could inhibit the growth of Xoo. Through Tn5 mutagenesis of P. taiwanensis, we showed that mutations in genes that encode components of the type VI secretion system (T6SS) as well as biosynthesis and maturation of pyoverdine resulted in reduced toxicity against Xoo. Our results indicated that T6SS is involved in the secretion of endogenous pyoverdine. Mutations in T6SS component genes affected the secretion of mature pyoverdine from the periplasmic space into the extracellular medium after pyoverdine precursor is transferred to the periplasm by the inner membrane transporter PvdE. In addition, we also showed that other export systems, i.e., the PvdRT-OpmQ and MexAB-OprM efflux systems (for which there have been previous suggestions of involvement) and the type II secretion system (T2SS), are not involved in pyoverdine secretion. PMID:27605490

  9. Disruption in Candida albicans of the TPS2 gene encoding trehalose-6-phosphate phosphatase affects cell integrity and decreases infectivity.

    PubMed

    Zaragoza, Oscar; de Virgilio, Claudio; Pontón, José; Gancedo, Carlos

    2002-05-01

    The gene CaTPS2 encoding trehalose-6-phosphate (T6P) phosphatase from Candida albicans has been cloned and disrupted in this organism. The Catps2/Catps2 mutant did not accumulate trehalose but accumulated high levels of T6P. Disruption of the two copies of the CaTPS2 gene did not abolish growth even at 42 degrees C, but decreased the growth rate. In the stationary phase, the Catps2/Catps2 mutant aggregated, more than 50% of its cells became permeable to propidium iodide and a large amount of protein was found in the culture medium. Aggregation occurred only at pH values higher than 7 and was avoided by osmoprotectants; it was never observed during the exponential phase of growth. The mutant formed colonies with a smooth border on Spider medium. Mice inoculated with 1.5 x 10(6) c.f.u. of wild-type cells died after 8 days, while 80% of those inoculated with the same number of c.f.u. of the Catps2/Catps2 mutant survived for at least 1 month. Reintroduction of the wild-type CaTPS2 gene in the Catps2/Catps2 mutant abolished the phenotypes described. It is hypothesized that the accumulation of T6P interferes with the assembly of a normal cell wall.

  10. Ultrasonic Impact Treatment to Improve Stress Corrosion Cracking Resistance of Welded Joints of Aluminum Alloy

    NASA Astrophysics Data System (ADS)

    Yu, J.; Gou, G.; Zhang, L.; Zhang, W.; Chen, H.; Yang, Y. P.

    2016-07-01

    Stress corrosion cracking is one of the major issues for welded joints of 6005A-T6 aluminum alloy in high-speed trains. High residual stress in the welded joints under corrosion results in stress corrosion cracking. Ultrasonic impact treatment was used to control the residual stress of the welded joints of 6005A-T6 aluminum alloy. Experimental tests show that ultrasonic impact treatment can induce compressive longitudinal and transverse residual stress in the welded joint, harden the surface, and increase the tensile strength of welded joints. Salt-fog corrosion tests were conducted for both an as-welded sample and an ultrasonic impact-treated sample. The surface of the treated sample had far fewer corrosion pits than that of the untreated sample. The treated sample has higher strength and lower tensile residual stress than the untreated sample during corrosion. Therefore, ultrasonic impact treatment is an effective technique to improve the stress corrosion cracking resistance of the welded joints of 6005A-T6 aluminum alloy.

  11. Chromatin condensation and recruitment of PHD finger proteins to histone H3K4me3 are mutually exclusive.

    PubMed

    Gatchalian, Jovylyn; Gallardo, Carmen Mora; Shinsky, Stephen A; Ospina, Ruben Rosas; Liendo, Andrea Mansilla; Krajewski, Krzysztof; Klein, Brianna J; Andrews, Forest H; Strahl, Brian D; M van Wely, Karel H; Kutateladze, Tatiana G

    2016-07-27

    Histone post-translational modifications, and specific combinations they create, mediate a wide range of nuclear events. However, the mechanistic bases for recognition of these combinations have not been elucidated. Here, we characterize crosstalk between H3T3 and H3T6 phosphorylation, occurring in mitosis, and H3K4me3, a mark associated with active transcription. We detail the molecular mechanisms by which H3T3ph/K4me3/T6ph switches mediate activities of H3K4me3-binding proteins, including those containing plant homeodomain (PHD) and double Tudor reader domains. Our results derived from nuclear magnetic resonance chemical shift perturbation analysis, orthogonal binding assays and cell fluorescence microscopy studies reveal a strong anti-correlation between histone H3T3/T6 phosphorylation and retention of PHD finger proteins in chromatin during mitosis. Together, our findings uncover the mechanistic rules of chromatin engagement for H3K4me3-specific readers during cell division. PMID:27016734

  12. Trehalose-6-phosphate synthesis controls yeast gluconeogenesis downstream and independent of SNF1.

    PubMed

    Deroover, Sofie; Ghillebert, Ruben; Broeckx, Tom; Winderickx, Joris; Rolland, Filip

    2016-06-01

    Trehalose-6-P (T6P), an intermediate of trehalose biosynthesis, was identified as an important regulator of yeast sugar metabolism and signaling. tps1Δ mutants, deficient in T6P synthesis (TPS), are unable to grow on rapidly fermentable medium with uncontrolled influx in glycolysis, depletion of ATP and accumulation of sugar phosphates. However, the exact molecular mechanisms involved are not fully understood. We show that SNF1 deletion restores the tps1Δ growth defect on glucose, suggesting that lack of TPS hampers inactivation of SNF1 or SNF1-regulated processes. In addition to alternative, non-fermentable carbon metabolism, SNF1 controls two major processes: respiration and gluconeogenesis. The tps1Δ defect appears to be specifically associated with deficient inhibition of gluconeogenesis, indicating more downstream effects. Consistently, Snf1 dephosphorylation and inactivation on glucose medium are not affected, as confirmed with an in vivo Snf1 activity reporter. Detailed analysis shows that gluconeogenic Pck1 and Fbp1 expression, protein levels and activity are not repressed upon glucose addition to tps1Δ cells, suggesting a link between the metabolic defect and persistent gluconeogenesis. While SNF1 is essential for induction of gluconeogenesis, T6P/TPS is required for inactivation of gluconeogenesis in the presence of glucose, downstream and independent of SNF1 activity and the Cat8 and Sip4 transcription factors. PMID:27189362

  13. Protein profiles and immunoreactivities of Acanthamoeba morphological groups and genotypes.

    PubMed

    Pumidonming, Wilawan; Koehsler, Martina; Leitsch, David; Walochnik, Julia

    2014-11-01

    Acanthamoeba is a free-living protozoan found in a wide variety of habitats. A classification of Acanthamoeba into currently eighteen genotypes (T1-T18) has been established, however, data on differences between genotypes on the protein level are scarce. The aim of this study was to compare protein and immunoreactivity profiles of Acanthamoeba genotypes. Thirteen strains, both clinical and non-clinical, from genotypes T4, T5, T6, T7, T9, T11 and T12, representing three morphological groups, were investigated for their protein profiles and IgG, IgM and IgA immunoreactivities. It was shown that protein and immunoreactivity profiles of Acanthamoeba genotypes T4, T5, T6, T7, T9, T11 and T12 are clearly distinct from each other, but the banding patterns correlate to the morphological groups. Normal human sera revealed anti-Acanthamoeba antibodies against isolates of all investigated genotypes, interestingly, however only very weak IgM and virtually no IgA immunoreactivity with T7 and T9, both representing morphological group I. The strongest IgG, IgM and IgA immunoreactivities were observed for genotypes T4, T5 and T6. Differences of both, protein and immunological patterns, between cytopathic and non-cytopathic strains, particularly within genotype T4, were not at the level of banding patterns, but rather in expression levels.

  14. New insights into virulence mechanisms of rice pathogen Acidovorax avenae subsp. avenae strain RS-1 following exposure to ß-lactam antibiotics

    PubMed Central

    Li, Bin; Ge, Mengyu; Zhang, Yang; Wang, Li; Ibrahim, Muhammad; Wang, Yanli; Sun, Guochang; Chen, Gongyou

    2016-01-01

    Recent research has shown that pathogen virulence can be altered by exposure to antibiotics, even when the growth rate is unaffected. Investigating this phenomenon provides new insights into understanding the virulence mechanisms of bacterial pathogens. This study investigates the phenotypic and transcriptomic responses of the rice pathogenic bacterium Acidovorax avenae subsp. avenae (Aaa) strain RS-1 to ß-lactam antibiotics especially Ampicillin (Amp). Our results indicate that exposure to Amp does not influence bacterial growth and biofilm formation, but alters the virulence, colonization capacity, composition of extracellular polymeric substances and secretion of Type VI secretion system (T6SS) effector Hcp. This attenuation in virulence is linked to unique or differential expression of known virulence-associated genes based on genome-wide transcriptomic analysis. The reliability of expression data generated by RNA-Seq was verified with quantitative real-time PCR of 21 selected T6SS genes, where significant down-regulation in expression of hcp gene, corresponding to the reduction in secretion of Hcp, was observed under exposure to Amp. Hcp is highlighted as a potential target for Amp, with similar changes observed in virulence-associated phenotypes between exposure to Amp and mutation of hcp gene. In addition, Hcp secretion is reduced in knockout mutants of 4 differentially expressed T6SS genes. PMID:26915352

  15. Prevalence of Trichinella spp. in black bears, grizzly bears, and wolves in the Dehcho Region, Northwest Territories, Canada, including the first report of T. nativa in a grizzly bear from Canada.

    PubMed

    Larter, Nicholas C; Forbes, Lorry B; Elkin, Brett T; Allaire, Danny G

    2011-07-01

    Samples of muscle from 120 black bears (Ursus americanus), 11 grizzly bears (Ursus arctos), and 27 wolves (Canis lupus) collected in the Dehcho Region of the Northwest Territories from 2001 to 2010 were examined for the presence of Trichinella spp. larvae using a pepsin-HCl digestion assay. Trichinella spp. larvae were found in eight of 11 (73%) grizzly bears, 14 of 27 (52%) wolves, and seven of 120 (5.8%) black bears. The average age of positive grizzly bears, black bears, and wolves was 13.5, 9.9, and approximately 4 yr, respectively. Larvae from 11 wolves, six black bears, and seven grizzly bears were genotyped. Six wolves were infected with T. nativa and five with Trichinella T6, four black bears were infected with T. nativa and two with Trichinella T6, and all seven grizzly bears were infected with Trichinella T6 and one of them had a coinfection with T. nativa. This is the first report of T. nativa in a grizzly bear from Canada. Bears have been linked to trichinellosis outbreaks in humans in Canada, and black bears are a subsistence food source for residents of the Dehcho region. In order to assess food safety risk it is important to monitor the prevalence of Trichinella spp. in both species of bear and their cohabiting mammalian food sources.

  16. Performance Verification of Impact Machines for Testing Plastics

    PubMed Central

    Siewert, T. A.; Vigliotti, D. P.; Dirling, L. B.; McCowan, C. N.

    1999-01-01

    Valid comparison of impact test energies reported by various organizations and over time depends on consistent performance of impact test machines. This paper investigates the influence of various specimen and test parameters on impact energies in the 1 J to 2 J range for both Charpy V-notch and Izod procedures, leading toward the identification of a suitable material for use in a program to verify machine performance. We investigated the influences on the absorbed energy of machine design, test material, specimen cross sectional area, and machine energy range. For comparison to published round robin data on common plastics, this study used some common metallic alloys, including those used in the international verification program for metals impact machines and in informal calibration programs of tensile machines. The alloys that were evaluated include AISI type 4340 steel, and five aluminum alloys: 2014-T6, 2024-T351, 2219-T87, 6061-T6, and 7075-T6. We found that certain metallic alloys have coefficients of variation comparable to those of the best plastics that are reported in the literature. Also, we found that the differences in absorbed energy between two designs of machines are smaller than the differences that can be attributed to the specimens alone.

  17. Prevalence of Trichinella spp. in black bears, grizzly bears, and wolves in the Dehcho Region, Northwest Territories, Canada, including the first report of T. nativa in a grizzly bear from Canada.

    PubMed

    Larter, Nicholas C; Forbes, Lorry B; Elkin, Brett T; Allaire, Danny G

    2011-07-01

    Samples of muscle from 120 black bears (Ursus americanus), 11 grizzly bears (Ursus arctos), and 27 wolves (Canis lupus) collected in the Dehcho Region of the Northwest Territories from 2001 to 2010 were examined for the presence of Trichinella spp. larvae using a pepsin-HCl digestion assay. Trichinella spp. larvae were found in eight of 11 (73%) grizzly bears, 14 of 27 (52%) wolves, and seven of 120 (5.8%) black bears. The average age of positive grizzly bears, black bears, and wolves was 13.5, 9.9, and approximately 4 yr, respectively. Larvae from 11 wolves, six black bears, and seven grizzly bears were genotyped. Six wolves were infected with T. nativa and five with Trichinella T6, four black bears were infected with T. nativa and two with Trichinella T6, and all seven grizzly bears were infected with Trichinella T6 and one of them had a coinfection with T. nativa. This is the first report of T. nativa in a grizzly bear from Canada. Bears have been linked to trichinellosis outbreaks in humans in Canada, and black bears are a subsistence food source for residents of the Dehcho region. In order to assess food safety risk it is important to monitor the prevalence of Trichinella spp. in both species of bear and their cohabiting mammalian food sources. PMID:21719845

  18. Comparative genome analyses of Serratia marcescens FS14 reveals its high antagonistic potential.

    PubMed

    Li, Pengpeng; Kwok, Amy H Y; Jiang, Jingwei; Ran, Tingting; Xu, Dongqing; Wang, Weiwu; Leung, Frederick C

    2015-01-01

    S. marcescens FS14 was isolated from an Atractylodes macrocephala Koidz plant that was infected by Fusarium oxysporum and showed symptoms of root rot. With the completion of the genome sequence of FS14, the first comprehensive comparative-genomic analysis of the Serratia genus was performed. Pan-genome and COG analyses showed that the majority of the conserved core genes are involved in basic cellular functions, while genomic factors such as prophages contribute considerably to genome diversity. Additionally, a Type I restriction-modification system, a Type III secretion system and tellurium resistance genes are found in only some Serratia species. Comparative analysis further identified that S. marcescens FS14 possesses multiple mechanisms for antagonism against other microorganisms, including the production of prodigiosin, bacteriocins, and multi-antibiotic resistant determinants as well as chitinases. The presence of two evolutionarily distinct Type VI secretion systems (T6SSs) in FS14 may provide further competitive advantages for FS14 against other microbes. To our knowledge, this is the first report of comparative analysis on T6SSs in the genus, which identifies four types of T6SSs in Serratia spp.. Competition bioassays of FS14 against the vital plant pathogenic bacterium Ralstonia solanacearum and fungi Fusarium oxysporum and Sclerotinia sclerotiorum were performed to support our genomic analyses, in which FS14 demonstrated high antagonistic activities against both bacterial and fungal phytopathogens.

  19. Type VI secretion system sheaths as nanoparticles for antigen display

    PubMed Central

    Del Tordello, Elena; Danilchanka, Olga; McCluskey, Andrew J.; Mekalanos, John J.

    2016-01-01

    The bacterial type 6 secretion system (T6SS) is a dynamic apparatus that translocates proteins between cells by a mechanism analogous to phage tail contraction. T6SS sheaths are cytoplasmic tubular structures composed of stable VipA-VipB (named for ClpV-interacting protein A and B) heterodimers. Here, the structure of the VipA/B sheath was exploited to generate immunogenic multivalent particles for vaccine delivery. Sheaths composed of VipB and VipA fused to an antigen of interest were purified from Vibrio cholerae or Escherichia coli and used for immunization. Sheaths displaying heterologous antigens generated better immune responses against the antigen and different IgG subclasses compared with soluble antigen alone. Moreover, antigen-specific antibodies raised against sheaths presenting Neisseria meningitidis factor H binding protein (fHbp) antigen were functional in a serum bactericidal assay. Our results demonstrate that multivalent nanoparticles based on the T6SS sheath represent a versatile scaffold for vaccine applications. PMID:26929342

  20. Precipitate microstructures and resulting properties of Al-Zn-Mg metal inert gas-weld heat-affected zones

    NASA Astrophysics Data System (ADS)

    Nicolas, M.; Deschamps, A.

    2004-05-01

    Using the combination of small-angle X-ray scattering (SAXS) and transmission electron microscopy (TEM), the precipitate microstructure is quantitatively investigated in the heat-affected zones (HAZs) of Al-Zn-Mg metal inert gas (MIG)-welds, and the resulting mechanical properties are determined by hardness measurements. Three initial states prior to welding (T4, T6, and T7) are investigated, and the subsequent microstructure evolution during natural aging and postwelding heat treatments (PWHTs) is assessed. The critical part of the HAZ is shown to be the transition region where partial dissolution of the initially present precipitates occurs. In this transition zone, precipitate coarsening is shown to occur for the T6 and T7 initial states, contrarily to the T4 material. After PWHT, the T6 and T7 materials experience a weak region related to this coarsening behavior, whereas the T4 material HAZ is able to recover a homogeneous microstructure after a suitably chosen PWHT. Simple model ramp heat treatments are shown to describe the main phenomena involved in the HAZ. Finally, a precipitation hardening model is successfully applied to the microstructural data to describe the hardness profiles in the various HAZs.

  1. Involvement of type VI secretion system in secretion of iron chelator pyoverdine in Pseudomonas taiwanensis.

    PubMed

    Chen, Wen-Jen; Kuo, Tzu-Yen; Hsieh, Feng-Chia; Chen, Pi-Yu; Wang, Chang-Sheng; Shih, Yu-Ling; Lai, Ying-Mi; Liu, Je-Ruei; Yang, Yu-Liang; Shih, Ming-Che

    2016-01-01

    Rice bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most destructive rice diseases worldwide. Therefore, in addition to breeding disease-resistant rice cultivars, it is desirable to develop effective biocontrol agents against Xoo. Here, we report that a soil bacterium Pseudomonas taiwanensis displayed strong antagonistic activity against Xoo. Using matrix-assisted laser desorption/ionization imaging mass spectrometry, we identified an iron chelator, pyoverdine, secreted by P. taiwanensis that could inhibit the growth of Xoo. Through Tn5 mutagenesis of P. taiwanensis, we showed that mutations in genes that encode components of the type VI secretion system (T6SS) as well as biosynthesis and maturation of pyoverdine resulted in reduced toxicity against Xoo. Our results indicated that T6SS is involved in the secretion of endogenous pyoverdine. Mutations in T6SS component genes affected the secretion of mature pyoverdine from the periplasmic space into the extracellular medium after pyoverdine precursor is transferred to the periplasm by the inner membrane transporter PvdE. In addition, we also showed that other export systems, i.e., the PvdRT-OpmQ and MexAB-OprM efflux systems (for which there have been previous suggestions of involvement) and the type II secretion system (T2SS), are not involved in pyoverdine secretion. PMID:27605490

  2. T346Hunter: A Novel Web-Based Tool for the Prediction of Type III, Type IV and Type VI Secretion Systems in Bacterial Genomes

    PubMed Central

    Martínez-García, Pedro Manuel; Ramos, Cayo; Rodríguez-Palenzuela, Pablo

    2015-01-01

    T346Hunter (Type Three, Four and Six secretion system Hunter) is a web-based tool for the identification and localisation of type III, type IV and type VI secretion systems (T3SS, T4SS and T6SS, respectively) clusters in bacterial genomes. Non-flagellar T3SS (NF-T3SS) and T6SS are complex molecular machines that deliver effector proteins from bacterial cells into the environment or into other eukaryotic or prokaryotic cells, with significant implications for pathogenesis of the strains encoding them. Meanwhile, T4SS is a more functionally diverse system, which is involved in not only effector translocation but also conjugation and DNA uptake/release. Development of control strategies against bacterial-mediated diseases requires genomic identification of the virulence arsenal of pathogenic bacteria, with T3SS, T4SS and T6SS being major determinants in this regard. Therefore, computational methods for systematic identification of these specialised machines are of particular interest. With the aim of facilitating this task, T346Hunter provides a user-friendly web-based tool for the prediction of T3SS, T4SS and T6SS clusters in newly sequenced bacterial genomes. After inspection of the available scientific literature, we constructed a database of hidden Markov model (HMM) protein profiles and sequences representing the various components of T3SS, T4SS and T6SS. T346Hunter performs searches of such a database against user-supplied bacterial sequences and localises enriched regions in any of these three types of secretion systems. Moreover, through the T346Hunter server, users can visualise the predicted clusters obtained for approximately 1700 bacterial chromosomes and plasmids. T346Hunter offers great help to researchers in advancing their understanding of the biological mechanisms in which these sophisticated molecular machines are involved. T346Hunter is freely available at http://bacterial-virulence-factors.cbgp.upm.es/T346Hunter. PMID:25867189

  3. Structures and stability of medium silicon clusters. II. Ab initio molecular orbital calculations of Si12-Si20

    NASA Astrophysics Data System (ADS)

    Zhu, X. L.; Zeng, X. C.; Lei, Y. A.; Pan, B.

    2004-05-01

    Ab initio all-electron molecular-orbital calculations are carried out to study the structures and relative stability of low-energy silicon clusters (Sin,n=12-20). Selected geometric isomers include those predicted by Ho et al. [Nature (London) 392, 582 (1998)] based on an unbiased search with tight-binding/genetic algorithm, as well as those found by Rata et al. [Phys. Rev. Lett. 85, 546 (2000)] based on density-functional tight-binding/single-parent evolution algorithm. These geometric isomers are optimized at the Møller-Plesset (MP2) MP2/6-31G(d) level. The single-point energy at the coupled-cluster single and double substitutions (including triple excitations) [CCSD(T)] CCSD(T)/6-31G(d) level for several low-lying isomers are further computed. Harmonic vibrational frequency analysis at the MP2/6-31G(d) level of theory is also undertaken to assure that the optimized geometries are stable. For Si12-Si17 and Si19 the isomer with the lowest-energy at the CCSD(T)/6-31G(d) level is the same as that predicted by Ho et al., whereas for Si18 and Si20, the same as predicted by Rata et al. However, for Si14 and Si15, the vibrational frequency analysis indicates that the isomer with the lowest CCSD(T)/6-31G(d) single-point energy gives rise to imaginary frequencies. Small structural perturbation onto the Si14 and Si15 isomers can remove the imaginary frequencies and results in new isomers with slightly lower MP2/6-31G(d) energy; however the new isomers have a higher single-point energy at the CCSD(T)/6-31G(d) level. For most Sin (n=12-18,20) the low-lying isomers are prolate in shape, whereas for Si19 a spherical-like isomer is slightly lower in energy at the CCSD(T)/6-31G(d) level than low-lying prolate isomers.

  4. Light weight and high strength materials made of recycled steel and aluminum

    NASA Astrophysics Data System (ADS)

    Nounezi, Thomas

    Recycling has proven not only to address today's economical, environmental and social issues, but also to be imperative for the sustainability of human technology. The current thesis has investigated the feasibility of a new philosophy for Recycling (Alloying-Recycling) using steel 1020 and aluminum 6061T6. The study was limited to the metallurgical aspects only and has highlighted the potential of recycled alloys made of recycled aluminum and steel to exhibit substantially increased wear resistance and strength-to-weight ratio as compared to initial primary materials. Three alloy-mixtures are considered: TN3 (5wt% 1020 +95wt% 6061T6); TN5 (0.7wt% 1020 + 99.3wt% 6061T6); and TN4 (10wt% 6061T6 + 90wt% 1020). A Tucker induction power supply system (3kW; 135-400 kHz) is used to melt the alloy mixtures for casting in graphite crucibles. Heat treatment of the cast samples is done using a radiation box furnace. Microscopy, Vickers hardness and pin-on-disc abrasive wear tests are performed. Casting destroyed the initial microstructures of the alloys leading to a hardness reduction in the as-cast and solution heat-treated aluminum rich samples to 60 Hv from 140 Hv. Ageing slightly increased the hardness of the cast samples and provided a wear resistance two times higher than that of the initial 6061T6 material. On the steel rich side, the hardness of the as-cast TN4 was 480 Hv, which is more than twice as high as the initial hardness of steel 1020 of 202 Hv; this hints to strong internal and residual stress, probably martensite formation during fast cooling following casting. Solution heat treatment lowered the hardness to the original value of steel 1020, but provided about ten (10) times higher wear resistance; this suggests higher ductility and toughness of normalised TN4 as compared to 1020. In addition, TN4 exhibits about 25% weight reduction as compared to 1020. The actual recycling process and the effect of non-metallic impurities shall be investigated in future

  5. Changes in exercise performance and hormonal concentrations over a big ten soccer season in starters and nonstarters.

    PubMed

    Kraemer, William J; French, Duncan N; Paxton, Nigel J; Häkkinen, Keijo; Volek, Jeff S; Sebastianelli, Wayne J; Putukian, Margot; Newton, Robert U; Rubin, Martyn R; Gómez, Ana L; Vescovi, Jason D; Ratamess, Nicholas A; Fleck, Steven J; Lynch, J Michael; Knuttgen, Howard G

    2004-02-01

    As a consequence of the physiological demands experienced during a competitive soccer season, the antagonistic relationship between anabolic and catabolic processes can affect performance. Twenty-five male collegiate soccer players were studied throughout a season (11 weeks) to investigate the effects of long-term training and competition. Subjects were grouped as starters (S; n = 11) and nonstarters (NS; n = 14). Measures of physical performance, body composition, and hormonal concentrations (testosterone [T] and cortisol [C]) were assessed preseason (T1) and 5 times throughout the season (T2-T6). Starters and NS participated in 83.06% and 16.95% of total game time, respectively. Nonstarters had a significant increase (+1.6%) in body fat at T6 compared to T1. Isokinetic strength of the knee extensors (1.05 rad.sec(-1)) significantly decreased in both S (-12%) and NS (-10%; p < or = 0.05) at T6. Significant decrements in sprint speed (+4.3%) and vertical jump (-13.8%) were found at T5 in S only. Though within normal ranges (10.4-41.6 nmol.L(-1)), concentrations of T at T1 were low for both groups, but increased significantly by T6. Concentrations of C were elevated in both groups, with concentrations at the high end of the normal range (normal range 138-635 nmol.L(-1)) at T1 and T4 in NS and T4 in S, with both groups remaining elevated at T6. Data indicate that players entering the season with low circulating concentrations of T and elevated levels of C can experience reductions in performance during a season, with performance decrements exacerbated in starters over nonstarters. Soccer players should therefore have a planned program of conditioning that does not result in an acute overtraining phenomenon prior to preseason (e.g., young players trying to get in shape quickly in the 6 to 8 weeks in the summer prior to reporting for preseason camp). The detrimental effects of inappropriate training do not appear to be unloaded during the season and catabolic

  6. miR-181b Promotes hepatic stellate cells proliferation by targeting p27 and is elevated in the serum of cirrhosis patients

    SciTech Connect

    Wang, Baocan; Li, Wenxi; Guo, Kun; Xiao, Yongtao; Wang, Yuqin; Fan, Jiangao

    2012-04-27

    Highlights: Black-Right-Pointing-Pointer miR-181a and miR-181b, especially, miR-181b could be induced by transforming growth factor-beta 1 (TGF-{beta}1) in hepatic stellate cells. Black-Right-Pointing-Pointer miR-181b could promote HSC-T6 cell proliferation by directly targeting the negative cell regulator-p27 in HSC-T6 cell. Black-Right-Pointing-Pointer miR-181b was identified as potential serum diagnostic marker for liver cirrhosis patients. -- Abstract: MicroRNAs, as a kind of negative gene regulators, were demonstrated to be involved in many types of diseases. In this study, we found that transforming growth factor-beta 1 could induce the expression of miR-181a and miR-181b, and miR-181b increased in the much higher folds than miR-181a. Because of the important role of transforming growth factor-beta 1 in HSC activation and liver cirrhosis, we investigate the effect of miR-181a and miR-181b on HSC proliferation. The results showed that miR-181b could promote HSC-T6 cell proliferation by regulating cell cycle. Further study showed p27, the cell cycle regulator, was the direct target of miR-181b in HSC-T6 cell. But miR-181a had no effects on HSC-T6 cell proliferation and cell cycle, and did not target p27. Interestingly, miR-181b is elevated significantly in serum of liver cirrhosis cases comparing to that of normal persons, whereas miR-181a expression was in the similar level with that of normal persons. These results suggested that miR-181b could be induced by TGF-{beta}1 and promote the growth of HSCs by directly targeting p27. The elevation of miR-181b in serum suggested that it may be potential diagnostic biomarkers for cirrhosis. As for miR-181a, it may work in TGF-{beta}1 pathway by a currently unknown mechanism.

  7. Chromium-free conversion coatings based on inorganic salts (Zr/Ti/Mn/Mo) for aluminum alloys used in aircraft applications

    NASA Astrophysics Data System (ADS)

    Santa Coloma, P.; Izagirre, U.; Belaustegi, Y.; Jorcin, J. B.; Cano, F. J.; Lapeña, N.

    2015-08-01

    Novel chromium-free conversion coatings based on Zr/Ti/Mn/Mo compounds were developed at a pilot scale to improve the corrosion resistance of the AA2024-T3 and AA7075-T6 aluminum alloys for aircraft applications. The influence of the presence of Zr and Ti in the Zr/Ti/Mn/Mo conversion bath's formulation on the corrosion resistance of the coated alloys was investigated. The corrosion resistance provided by the conversion coatings was evaluated by salt spray exposure and potentiodynamic sweeps. Optical and scanning electron microscopy coupled with energy dispersive spectroscopy (SEM/EDS) and atomic force microscopy (AFM) operating in the Kelvin Probe mode (SKPFM) were used to provide microstructural information of the coated samples that achieved the best results in the corrosion tests. The salt spray test evidenced the higher corrosion resistance of the coated samples compared to the bare surfaces for both alloys. The potentiodynamic tests showed that the corrosion current density decreased for coated AA7075-T6 and AA2024-T3 alloys, which indicated an obvious improvement of the corrosion resistance with all the processes for both alloys. Although the corrosion resistance of the coated samples appeared to be higher for the alloy AA7075-T6 than for the alloy AA2024-T3, both alloys achieved the best corrosion protection with the coatings deposited from conversion bath formulations containing no titanium salts. The microscopy analysis on the coated AA7075-T6 samples revealed that a local deposition of Zr compounds and, possibly, an oxidation process occurred in the vicinity of the alloy's intermetallic particles. The amount of the Zr deposits at these locations increased with coating's formulations without Ti, which provided the best corrosion resistance. The Cr-free conversion coatings developed in this study for the AA7075-T6 and AA2024-T3 alloys do not meet yet the strict requirements of the aircraft industry. However, they significantly improved the corrosion

  8. Correlations between lactate and ventilatory thresholds and the maximal lactate steady state in elite cyclists.

    PubMed

    Van Schuylenbergh, R; Vanden Eynde, B; Hespel, P

    2004-08-01

    We investigated the validity of different lactate and ventilatory threshold methods, to estimate heart rate and power output corresponding with the maximal lactate steady-state (MLSS) in elite cyclists. Elite cyclists (n = 21; 21 +/- 0.4 y; VO2peak, 5.4 +/- 0.2 l x min (-1)) performed either one (n = 10) or two (n = 11) maximal graded exercise tests, as well as two to three 30-min constant-load tests to determine MLSS, on their personal race bicycle which was mounted on an ergometer. Initial workload for the graded tests was 100 Watt and was increased by either 5 % of body mass (in Watt) with every 30 s (T30 s), or 60 % of body mass (in Watt) with every 6 min (T6min). MLSS was defined as the highest constant workload during which lactate increased no more than 1 mmol x l (-1) from min 10 to 30. In T30 s and T6 min the 4 mmol (TH-La4), the Conconi (TH-Con) and dmax (TH-Dm) lactate threshold were determined. The dmax lactate threshold was defined as the point that yields the maximal distance from the lactate curve to the line formed by the lowest and highest lactate values of the curve. In T30 s also ventilatory (TH-Ve) and Vslope (TH-Vs) thresholds were calculated. Time to exhaustion was 36 +/- 1 min for T30 s versus 39 +/- 1 min for T6 min. None of the threshold measures in T30 s, except TH-Vs (r2 = 0.77 for heart rate) correlated with either MLSS heart rate or power output. During T6 min, power output at TH-Dm was closely correlated with MLSS power (r2=0.72). Low correlations were found between MLSS heart rate and heart rate measured at TH-Dm (r2=0.46) and TH-La4 (r2=0.25), respectively, during T6 min. It is concluded that it is not possible to precisely predict heart rate or power output corresponding with MLSS in elite cyclists, from a single graded exercise test causing exhaustion within 35-40 min. The validity of MLSS predicted from an incremental test must be verified by a 30-min constant-load test.

  9. [Effects of mulching on soil moisture in a dryland winter wheat field, Northwest China].

    PubMed

    Fan, Ying-Dan; Chai, Shou-Xi; Cheng, Hong-Bo; Chen, Yu-Zhang; Yang, Chang-Gang; Huang, Cai-Xia; Chang, Lei; Pang, Lei

    2013-11-01

    This paper studied the effects of different mulching modes on the soil moisture in a semi-arid rainfed area of Loess Plateau, Northwest China. Seven treatments were installed, i. e., mulching plastic film in summer (T1), mulching plastic film in autumn (T2), mulching 5 cm long wheat straw in summer (T3), mulching whole wheat straw in summer (T4), mulching plastic film in summer plus wheat straw (T5), mulching used plastic film after harvest (T6), and un-mulching (CK). In T6, the soil moisture in different layers at different crop growth stages was all higher than that in CK. In the other five mulching treatments, the soil moisture in 0-90 cm layer before flowering stage was obviously higher, but that in 0-90 cm layer after flowering stage and in 90-200 cm layer during the whole growth season was lower than that of CK. The soil moisture in 0-200 cm layer in T6 during the whole growth period was significantly higher than that in CK, with a difference of 0.9%, but the soil moisture in 0-200 cm layer in other mulching treatments was lower. As compared with plastic film mulching, straw mulching increased the soil moisture in 0-200 cm layer. The soil moisture under mulching with used plastic film after harvest was higher than that under mulching with new plastic film. As compared to CK, the grain yield of winter wheat with plastic film mulching was increased by 20.3%-29.0%, and that With straw mulching was increased by 5.0%-16.7%. There was a significant positive correlation between the crop productivity and the soil water consumption during the growth period (r = 0.77*).

  10. On the shaping factors of the secondary microseismic wavefield

    NASA Astrophysics Data System (ADS)

    Gualtieri, L.; Stutzmann, E.; Capdeville, Y.; Farra, V.; Mangeney, A.; Morelli, A.

    2015-09-01

    Seismic noise in the period band 3-10 s is known as secondary microseism, and it is generated at the ocean surface by the interaction of ocean gravity waves coming from nearly opposite directions. In this paper, we investigate the seismic content of the wavefield generated by a source at the ocean surface and three of the major wavefield shaping factors using the 2-D spectral element method: the ocean-continent boundary, the source site effect, and the thickness of seafloor sediments. The seismic wavefield recorded on the vertical component seismograms below the seafloor is mainly composed of the fundamental mode and the first overtone of Rayleigh waves. A mode conversion from the first overtone to the fundamental mode of Rayleigh waves occurs at the ocean-continent boundary. The presence of a continental shelf at the ocean-continent boundary produces a negligible effect on land-recorded seismograms, whereas the source site effect, i.e., the source location with respect to the local ocean depth and sediment thickness, plays the major role. A source in shallow water mostly enhances the fundamental mode of Rayleigh waves, whereas a source in deep water mainly enhances the first overtone of Rayleigh waves. Land-recorded long-period signals (T > 6 s) are mostly due to deep water sources, whereas land-recorded short-period signals (T < 6 s) are due to sources in relatively shallow water, located close to the shelf break. Seafloor sediments around the source region trap seismic waves reducing the amplitude of land-recorded signals, especially at long periods (T > 6 s).

  11. Lymphoid and other tissue-specific phenotypes of polyomavirus enhancer recombinants: positive and negative combinational effects on enhancer specificity and activity.

    PubMed Central

    Campbell, B A; Villarreal, L P

    1986-01-01

    Heterologous enhancer recombinants and deletions of the polyomavirus (Py) noncoding region were constructed and analyzed for tissue specificity of DNA replication and transcription in a number of lymphoid and other cell lines. The simian virus 40 72-base-pair repeat, mouse immunoglobulin heavy-chain enhancer, and Moloney murine leukemia virus enhancer were inserted into the PvuII-D locus (nucleotides 5128 through 5265) of Py. The ability of these recombinants and the parental PvuII-D deletion mutant to replicate in permissive 3T6 cells and MOP-6 cells as well as in nonpermissive mouse B lymphoid, T lymphoid, mastocyte, and embryonal carcinoma cells was determined. Wild-type Py DNA was not permissive for replication in most lymphoid cell lines, except one hybridoma line. Simply deleting the Py PvuII-D region, however, gave Py an expanded host range, allowing high-level replication in some T lymphoid and mastocytoma cell lines, indicating that this element can be a tissue-specific negative as well as positive element. Substitution of the murine leukemia virus enhancer for Py PvuII-D yielded a Py genome which retained the ability to replicate in 3T6 cells but also replicated well in B lymphoid cells. Substitution with the immunoglobulin heavy-chain enhancer allowed replication in B lymphoid cells but interfered with replication in 3T6 cells and mastocytomas. Surprisingly, substitution with the simian virus 40 72-base-pair enhancer repeat gave a recombinant which would not replicate in any cell line tried, including MOP-6 cells, even though other recombinants with this enhancer would replicate. Thus, we observed both cooperation and interference in these combinations between enhancer components and the Py genome and that these combined activities were cell specific. These results are presented as evidence that there may be a positional dependence, or syntax, for the recognition of genetic elements controlling Py tissue specificity. Images PMID:3023917

  12. Colistin-resistant, lipopolysaccharide-deficient Acinetobacter baumannii responds to lipopolysaccharide loss through increased expression of genes involved in the synthesis and transport of lipoproteins, phospholipids, and poly-β-1,6-N-acetylglucosamine.

    PubMed

    Henry, Rebekah; Vithanage, Nuwan; Harrison, Paul; Seemann, Torsten; Coutts, Scott; Moffatt, Jennifer H; Nation, Roger L; Li, Jian; Harper, Marina; Adler, Ben; Boyce, John D

    2012-01-01

    We recently demonstrated that colistin resistance in Acinetobacter baumannii can result from mutational inactivation of genes essential for lipid A biosynthesis (Moffatt JH, et al., Antimicrob. Agents Chemother. 54:4971-4977). Consequently, strains harboring these mutations are unable to produce the major Gram-negative bacterial surface component, lipopolysaccharide (LPS). To understand how A. baumannii compensates for the lack of LPS, we compared the transcriptional profile of the A. baumannii type strain ATCC 19606 to that of an isogenic, LPS-deficient, lpxA mutant strain. The analysis of the expression profiles indicated that the LPS-deficient strain showed increased expression of many genes involved in cell envelope and membrane biogenesis. In particular, upregulated genes included those involved in the Lol lipoprotein transport system and the Mla-retrograde phospholipid transport system. In addition, genes involved in the synthesis and transport of poly-β-1,6-N-acetylglucosamine (PNAG) also were upregulated, and a corresponding increase in PNAG production was observed. The LPS-deficient strain also exhibited the reduced expression of genes predicted to encode the fimbrial subunit FimA and a type VI secretion system (T6SS). The reduced expression of genes involved in T6SS correlated with the detection of the T6SS-effector protein AssC in culture supernatants of the A. baumannii wild-type strain but not in the LPS-deficient strain. Taken together, these data show that, in response to total LPS loss, A. baumannii alters the expression of critical transport and biosynthesis systems associated with modulating the composition and structure of the bacterial surface. PMID:22024825

  13. An eighteen-month follow-up study on the effects of Intravitreal Dexamethasone Implant in diabetic macular edema refractory to anti-VEGF therapy

    PubMed Central

    Pacella, Fernanda; Romano, Maria Rosaria; Turchetti, Paolo; Tarquini, Giovanna; Carnovale, Anna; Mollicone, Antonella; Mastromatteo, Alessandra; Pacella, Elena

    2016-01-01

    AIM To evaluate the long-term efficacy and safety of dexamethasone implants in subjects affected by diabetic macular edema (DME) resistant to anti-vascular endothelial growth factor (VEGF) therapy. METHODS Thirty-two DME patients were enrolled. A 700 microgram slow release Intravitreal Dexamethasone Implant (Ozurdex®) was placed in the vitreous cavity. All patients were followed for 18mo. Best-corrected visual acuity (BCVA) measured with Early Treatment Diabetic Retinopathy Study (ETDRS) and central macular thickness (CMT) exams were carried out at baseline (T0) and after 1 (T1), 3 (T3), 4 (T4), 6 (T6), 9 (T9), 12 (T12), 15 (T15), and 18mo (T18) post injection. RESULTS Repeated measures ANOVA showed an effect of treatment on ETDRS (P<0.0001). Post hoc analyses revealed that ETDRS values were significantly increased at T1, T3, T4, T9, and T15 (P<0.001) as compared to baseline value (T0). At T6, T12, and T18, ETDRS values were still statistically higher than baseline (P<0.001 vs T0). However, at these time points, we observed a trend to return to baseline conditions. ANOVA also showed an effect of treatment (P<0.0001). CMT decreased significantly at T1, T3, T4, T9, and T15 (P<0.001). At T6 (P<0.01), T12 and T18 (P<0.001) CMT was also significantly lower than T0 although a trend to return to the baseline conditions was also observed. CONCLUSION Our findings demonstrate that Intravitreal Dexamethasone Implant is a good option to improve BCVA and CMT in DME patients resistant to anti-VEGF therapy. Our data also show that the use of drugs administered directly into the vitreous allows achieving appropriate and long-lasting concentration at the site of disease without systemic side effects. PMID:27803859

  14. Increased thermal and osmotic stress resistance in Listeria monocytogenes 568 grown in the presence of trehalose due to inactivation of the phosphotrehalase-encoding gene treA.

    PubMed

    Ells, Timothy C; Truelstrup Hansen, Lisbeth

    2011-10-01

    The food-borne pathogen Listeria monocytogenes is a problem for food processors and consumers alike, as the organism is resistant to harsh environmental conditions and inimical barriers implemented to prevent the survival and/or growth of harmful bacteria. One mechanism by which listeriae mediate survival is through the accumulation of compatible solutes, such as proline, betaine and carnitine. In other bacteria, including Escherichia coli, the synthesis and accumulation of another compatible solute, trehalose, are known to aid in the survival of stressed cells. The objective of this research was to investigate trehalose metabolism in L. monocytogenes, where the sugar is thought to be transferred across the cytoplasmic membrane via a specific phosphoenolpyruvate phosphotransferase system and phosphorylation to trehalose-6-phosphate (T6P). The latter is subsequently broken down into glucose and glucose-6-phosphate by α,α-(1,1) phosphotrehalase, the putative product of the treA gene. Here we report on an isogenic treA mutant of L. monocytogenes 568 (568:ΔTreA) which, relative to the wild-type strain, displays increased tolerances to multiple stressors, including heat, high osmolarity, and desiccation. This is the first study to examine the putative trehalose operon in L. monocytogenes, and we demonstrate that lmo1254 (treA) in L. monocytogenes 568 indeed encodes a phosphotrehalase required for the hydrolysis of T6P. Disruption of the treA gene results in the accumulation of T6P which is subsequently dephosphorylated to trehalose in the cytosol, thereby contributing to the stress hardiness observed in the treA mutant. This study highlights the importance of compatible solutes for microbial survival in adverse environments.

  15. The Crystal Structure and Small-Angle X-Ray Analysis of CsdL/TcdA Reveal a New tRNA Binding Motif in the MoeB/E1 Superfamily

    PubMed Central

    López-Estepa, Miguel; Ardá, Ana; Savko, Martin; Round, Adam; Shepard, William E.; Bruix, Marta; Coll, Miquel; Fernández, Francisco J.; Jiménez-Barbero, Jesús; Vega, M. Cristina

    2015-01-01

    Cyclic N6-threonylcarbamoyladenosine (‘cyclic t6A’, ct6A) is a non-thiolated hypermodification found in transfer RNAs (tRNAs) in bacteria, protists, fungi and plants. In bacteria and yeast cells ct6A has been shown to enhance translation fidelity and efficiency of ANN codons by improving the faithful discrimination of aminoacylated tRNAs by the ribosome. To further the understanding of ct6A biology we have determined the high-resolution crystal structures of CsdL/TcdA in complex with AMP and ATP, an E1-like activating enzyme from Escherichia coli, which catalyzes the ATP-dependent dehydration of t6A to form ct6A. CsdL/TcdA is a dimer whose structural integrity and dimer interface depend critically on strongly bound K+ and Na+ cations. By using biochemical assays and small-angle X-ray scattering we show that CsdL/TcdA can associate with tRNA with a 1:1 stoichiometry and with the proper position and orientation for the cyclization of t6A. Furthermore, we show by nuclear magnetic resonance that CsdL/TcdA engages in transient interactions with CsdA and CsdE, which, in the latter case, involve catalytically important residues. These short-lived interactions may underpin the precise channeling of sulfur atoms from cysteine to CsdL/TcdA as previously characterized. In summary, the combination of structural, biophysical and biochemical methods applied to CsdL/TcdA has afforded a more thorough understanding of how the structure of this E1-like enzyme has been fine tuned to accomplish ct6A synthesis on tRNAs while providing support for the notion that CsdA and CsdE are able to functionally interact with CsdL/TcdA. PMID:25897750

  16. Reconstitution and characterization of eukaryotic N6-threonylcarbamoylation of tRNA using a minimal enzyme system

    PubMed Central

    Wan, Leo C. K.; Mao, Daniel Y. L.; Neculai, Dante; Strecker, Jonathan; Chiovitti, David; Kurinov, Igor; Poda, Gennadiy; Thevakumaran, Neroshan; Yuan, Fang; Szilard, Rachel K.; Lissina, Elena; Nislow, Corey; Caudy, Amy A.; Durocher, Daniel; Sicheri, Frank

    2013-01-01

    The universally conserved Kae1/Qri7/YgjD and Sua5/YrdC protein families have been implicated in growth, telomere homeostasis, transcription and the N6-threonylcarbamoylation (t6A) of tRNA, an essential modification required for translational fidelity by the ribosome. In bacteria, YgjD orthologues operate in concert with the bacterial-specific proteins YeaZ and YjeE, whereas in archaeal and eukaryotic systems, Kae1 operates as part of a larger macromolecular assembly called KEOPS with Bud32, Cgi121, Gon7 and Pcc1 subunits. Qri7 orthologues function in the mitochondria and may represent the most primitive member of the Kae1/Qri7/YgjD protein family. In accordance with previous findings, we confirm that Qri7 complements Kae1 function and uncover that Qri7 complements the function of all KEOPS subunits in growth, t6A biosynthesis and, to a partial degree, telomere maintenance. These observations suggest that Kae1 provides a core essential function that other subunits within KEOPS have evolved to support. Consistent with this inference, Qri7 alone is sufficient for t6A biosynthesis with Sua5 in vitro. In addition, the 2.9 Å crystal structure of Qri7 reveals a simple homodimer arrangement that is supplanted by the heterodimerization of YgjD with YeaZ in bacteria and heterodimerization of Kae1 with Pcc1 in KEOPS. The partial complementation of telomere maintenance by Qri7 hints that KEOPS has evolved novel functions in higher organisms. PMID:23620299

  17. Temperature effects on the deformation and fracture of Al-Li-Cu-In alloys

    NASA Technical Reports Server (NTRS)

    Wagner, John A.; Gangloff, Richard P.

    1991-01-01

    The crack initiation and growth fracture resistance of Al-Cu-Li and Al-Cu-Li-In alloys were characterized and optimized for cryogenic tank applications. Presently, the effects of stress state and temperature is being determined on the fracture toughness and fracture mechanisms of commercially available Vintage 3 2090-T81 and experimental 2090+In-T6. Precracked J-integral specimens of both alloys were tested at ambient and cryogenic temperatures in the plane stress and plane strain conditions. Considering ambient temperature, results showed that 2090-T81 exhibited the highest toughness in both plane strain and plane stress conditions. For the plane strain condition, reasonable crack initiation and growth toughness of 1090-T81 are associated with a significant amount of delamination and transgranular fracture. Plane stress toughnesses were higher and fracture was characterized by shear cracking with minimal delaminations. In comparisons, the fracture behavior of 2090+In-T6 is significantly degraded by subgrain boundary precipitation. Toughness is low and characterized by intersubgranular fracture with no delamination in the plane stress or plane strain conditions. Intersubgranular cracking is a low energy event which presumably occurs prior to the onset of slip band cracking. Copious grain boundary precipitation is atypical of commercially available 2090. At cryogenic temperatures, both alloys exhibit increased yield strength, toughness, and amount of delamination and shear cracking. The change in fracture mode of 2090+In-T6 from intersubgranular cracking at ambient temperature to a combination of intersubgranular cracking, shear cracking, and delamination at cryogenic temperature is the subject of further investigation.

  18. Small and middle T antigens contribute to lytic and abortive polyomavirus infection

    SciTech Connect

    Tuerler, H.; Salomon, C.

    1985-02-01

    Using three different polyomavirus hr-t mutants and two polyomavirus mlT mutants, the authors studied induction of S-phase by mutants and wild-type virus in quiescent mouse kidney cells, mouse 3T6 cells, and FR 3T3 cells. At different times after infection, they measured the proportion of T-antigen-positive cells, the incorporation of (/sup 3/H)thymidine, the proportion of DNA-synthesizing cells, and the increase in total DNA, RNA, and protein content of the cultures. In permissive mouse cells, they also determined the amount of viral DNA and the proportion of viral capsid-producing cells. In polyomavirus hr-t mutant-infected cultures, the onset of host DNA replication was delayed by several hours, and a smaller proportion of T-antigen-positive cells entered S-phase than in wild-type-infected cultures. Of the two polyomavirus mlT mutants studied, dl-23 behaved similarly to wild-type virus in many, but not all, parameters tested. The poorly replicating but well-transforming mutant dl-8 was able to induce S-phase, and (in permissive cells) progeny virus production, in only about one-third of the T-antigen-positive cells. From the experiments, the authors concluded that mutations affecting small and middle T-antigen cause a reduction in the proportion of cells responding to virus infection and a prolongation of the early phase, i.e., the period before cells center S-phase. In hr-t mutant-infected mouse 3T6 cells, production of viral DNA was <10% of that in wild-type-infected cultures; low hr-t progeny production in 3T6 cells was therefore largely due to poor viral DNA replication.

  19. Two-component PhoB-PhoR regulatory system and ferric uptake regulator sense phosphate and iron to control virulence genes in type III and VI secretion systems of Edwardsiella tarda.

    PubMed

    Chakraborty, Smarajit; Sivaraman, J; Leung, Ka Yin; Mok, Yu-Keung

    2011-11-11

    Inorganic phosphate (P(i)) and iron are essential nutrients that are depleted by vertebrates as a protective mechanism against bacterial infection. This depletion, however, is sensed by some pathogens as a signal to turn on the expression of virulence genes. Here, we show that the PhoB-PhoR two-component system senses changes in P(i) concentration, whereas the ferric uptake regulator (Fur) senses changes in iron concentration in Edwardsiella tarda PPD130/91 to regulate the expression of type III and VI secretion systems (T3SS and T6SS) through an E. tarda secretion regulator, EsrC. In sensing low P(i) concentration, PhoB-PhoR autoregulates and activates the phosphate-specific transport operon, pstSCAB-phoU, by binding directly to the Pho box in the promoters of phoB and pstS. PhoB also binds with EsrC simultaneously on the promoter of an E. tarda virulence protein, evpA, to regulate directly the transcription of genes from T6SS. In addition, PhoB requires and interacts with PhoU to activate esrC and suppress fur indirectly through unidentified regulators. Fur, on the other hand, senses high iron concentration and binds directly to the Fur box in the promoter of evpP to inhibit EsrC binding to the same region. In addition, Fur suppresses transcription of phoB, pstSCAB-phoU, and esrC indirectly via unidentified regulators, suggesting negative cross-talk with the Pho regulon. Physical interactions exist between Fur and PhoU and between Fur and EsrC. Our findings suggest that T3SS and T6SS may carry out distinct roles in the pathogenicity of E. tarda by responding to different environmental factors.

  20. MLL-AF6 fusion oncogene sequesters AF6 into the nucleus to trigger RAS activation in myeloid leukemia.

    PubMed

    Manara, Elena; Baron, Emma; Tregnago, Claudia; Aveic, Sanja; Bisio, Valeria; Bresolin, Silvia; Masetti, Riccardo; Locatelli, Franco; Basso, Giuseppe; Pigazzi, Martina

    2014-07-10

    A rare location, t(6;11)(q27;q23) (MLL-AF6), is associated with poor outcome in childhood acute myeloid leukemia (AML). The described mechanism by which MLL-AF6, through constitutive self-association and in cooperation with DOT-1L, activates aberrant gene expression does not explain the biological differences existing between t(6;11)-rearranged and other MLL-positive patients nor their different clinical outcome. Here, we show that AF6 is expressed in the cytoplasm of healthy bone marrow cells and controls rat sarcoma viral oncogene (RAS)-guanosine triphosphate (GTP) levels. By contrast, in MLL-AF6-rearranged cells, AF6 is found localized in the nucleus, leading to aberrant activation of RAS and of its downstream targets. Silencing MLL-AF6, we restored AF6 localization in the cytoplasm, thus mediating significant reduction of RAS-GTP levels and of cell clonogenic potential. The rescue of RAS-GTP levels after MLL-AF6 and AF6 co-silencing confirmed that MLL-AF6 oncoprotein potentiates the activity of the RAS pathway through retention of AF6 within the nucleus. Exposure of MLL-AF6-rearranged AML blasts to tipifarnib, a RAS inhibitor, leads to cell autophagy and apoptosis, thus supporting RAS targeting as a novel potential therapeutic strategy in patients carrying t(6;11). Altogether, these data point to a novel role of the MLL-AF6 chimera and show that its gene partner, AF6, is crucial in AML development.

  1. Reconstitution and characterization of eukaryotic N6-threonylcarbamoylation of tRNA using a minimal enzyme system.

    PubMed

    Wan, Leo C K; Mao, Daniel Y L; Neculai, Dante; Strecker, Jonathan; Chiovitti, David; Kurinov, Igor; Poda, Gennadiy; Thevakumaran, Neroshan; Yuan, Fang; Szilard, Rachel K; Lissina, Elena; Nislow, Corey; Caudy, Amy A; Durocher, Daniel; Sicheri, Frank

    2013-07-01

    The universally conserved Kae1/Qri7/YgjD and Sua5/YrdC protein families have been implicated in growth, telomere homeostasis, transcription and the N6-threonylcarbamoylation (t(6)A) of tRNA, an essential modification required for translational fidelity by the ribosome. In bacteria, YgjD orthologues operate in concert with the bacterial-specific proteins YeaZ and YjeE, whereas in archaeal and eukaryotic systems, Kae1 operates as part of a larger macromolecular assembly called KEOPS with Bud32, Cgi121, Gon7 and Pcc1 subunits. Qri7 orthologues function in the mitochondria and may represent the most primitive member of the Kae1/Qri7/YgjD protein family. In accordance with previous findings, we confirm that Qri7 complements Kae1 function and uncover that Qri7 complements the function of all KEOPS subunits in growth, t(6)A biosynthesis and, to a partial degree, telomere maintenance. These observations suggest that Kae1 provides a core essential function that other subunits within KEOPS have evolved to support. Consistent with this inference, Qri7 alone is sufficient for t(6)A biosynthesis with Sua5 in vitro. In addition, the 2.9 Å crystal structure of Qri7 reveals a simple homodimer arrangement that is supplanted by the heterodimerization of YgjD with YeaZ in bacteria and heterodimerization of Kae1 with Pcc1 in KEOPS. The partial complementation of telomere maintenance by Qri7 hints that KEOPS has evolved novel functions in higher organisms.

  2. Influence of Aging Treatments on Alterations of Microstructural Features and Stress Corrosion Cracking Behavior of an Al-Zn-Mg Alloy

    NASA Astrophysics Data System (ADS)

    Rout, Prasanta Kumar; Ghosh, M. M.; Ghosh, K. S.

    2015-07-01

    7xxx series Al-Zn-Mg-(Cu) alloys have higher strength in their peak-aged (T6) states compared with other age-hardenable aluminum alloys; however, the maximum strength peak-aged state is more susceptible to stress corrosion cracking (SCC) which leads to catastrophic failure. The over-aged (T7) temper with 10-15% lower strength has higher resistance to SCC requiring oversized structural aerospace component applications. The medium-strength AA7017 Al-Zn-Mg weldable alloy without Cu is also prone to SCC under certain environmental conditions. In the present investigation, the SCC behaviors of an AA7017 Al-Zn-Mg alloys of different tempers have been assessed. Specific aging schedules have been adapted to an AA7017 alloy to produce various tempers, e.g., under-, peak-(T6), over-(T7), and highly over-aged tempers. Artificial aging behavior of the AA7017 alloy has been characterized by hardness, electrical conductivity measurements, x-ray diffraction, differential scanning calorimetry, and electrochemical studies. Slow strain rate test technique was used to assess the SCC behaviors of the AA7017 alloys of under-, T6, T7, and highly over-aged tempers in 3.5 wt.% NaCl solution at free corrosion potential (FCP) and at applied anodic potential, as well. Results revealed that the AA7017 alloy tempers are not susceptible to SCC in 3.5 wt.% NaCl solution at FCP, but severely damaging to SCC at applied anodic potentials. Microstructural features, showing a non-recrystallized grain structure and the presence of discrete, widely spaced, not-interconnected η precipitates at the grain boundaries, are the contributive factors by virtue of which the alloy tempers at FCP did not exhibit SCC. However, the applied anodic potential resulted in rapid metal dissolution from the grain boundary region and led to SCC. The local anodic dissolution (LAD) is believed to be the associated SCC mechanism.

  3. Zinc transporter gene expression and glycemic control in post-menopausal women with Type 2 diabetes mellitus.

    PubMed

    Foster, Meika; Chu, Anna; Petocz, Peter; Samman, Samir

    2014-10-01

    Type 2 diabetes mellitus (DM) is associated often with underlying zinc deficiency and nutritional supplements such as zinc may be of therapeutic benefit in the disease. In a randomized, double-blind, placebo-controlled, 12-week trial in postmenopausal women (n=48) with Type 2 DM we investigated the effects of supplementation with zinc (40mg/d) and flaxseed oil (FSO; 2g/d) on the gene expression of zinc transporters (ZnT1, ZnT5, ZnT6, ZnT7, ZnT8, Zip1, Zip3, Zip7, and Zip10) and metallothionein (MT-1A, and MT-2A), and markers of glycemic control (glucose, insulin, glycosylated hemoglobin [HbA1c]). The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. No significant effects of zinc or FSO supplementation were observed on glycemic marker concentrations, HOMA-IR or fold change over 12 weeks in zinc transporter and metallothionein gene expression. In multivariate analysis, the change over 12 weeks in serum glucose concentrations (P=0.001) and HOMA-IR (P=0.001) predicted the fold change in Zip10. In secondary analysis, marginal statistical significance was observed with the change in both serum glucose concentrations (P=0.003) and HOMA-IR (P=0.007) being predictive of the fold change in ZnT6. ZnT8 mRNA expression was variable; HbA1c levels were higher (P=0.006) in participants who exhibited ZnT8 expression compared to those who did not. The significant predictive relationships between Zip10, ZnT6, serum glucose and HOMA-IR are preliminary, as is the relationship between HbA1c and ZnT8; nevertheless the observations support an association between Type 2 DM and zinc homeostasis that requires further exploration.

  4. Anxious or Depressed and Still Happy?

    PubMed Central

    Spinhoven, Philip; Elzinga, Bernet M.; Giltay, Erik; Penninx, Brenda W. J. H.

    2015-01-01

    This study aimed to examine cross-sectionally to what extent persons with higher symptom levels or a current or past emotional disorder report to be less happy than controls and to assess prospectively whether time-lagged measurements of extraversion and neuroticism predict future happiness independent of time-lagged measurements of emotional disorders or symptom severity. A sample of 2142 adults aged 18–65, consisting of healthy controls and persons with current or past emotional disorder according to DSM-IV criteria completed self-ratings for happiness and emotional well-being and symptom severity. Lagged measurements of personality, symptom severity and presence of anxiety and depressive disorder at T0 (year 0), T2 (year 2) and T4 (year 4) were used to predict happiness and emotional well-being at T6 (year 6) controlling for demographics. In particular persons with more depressive symptoms, major depressive disorder, social anxiety disorder and comorbid emotional disorders reported lower levels of happiness and emotional well-being. Depression symptom severity and to a lesser extent depressive disorder predicted future happiness and emotional well-being at T6. Extraversion and to a lesser extent neuroticism also consistently forecasted future happiness and emotional well-being independent of concurrent lagged measurements of emotional disorders and symptoms. A study limitation is that we only measured happiness and emotional well-being at T6 and our measures were confined to hedonistic well-being and did not include psychological and social well-being. In sum, consistent with the two continua model of emotional well-being and mental illness, a ‘happy’ personality characterized by high extraversion and to a lesser extent low neuroticism forecasts future happiness and emotional well-being independent of concurrently measured emotional disorders or symptom severity levels. Boosting positive emotionality may be an important treatment goal for persons

  5. The crystal structure and small-angle X-ray analysis of CsdL/TcdA reveal a new tRNA binding motif in the MoeB/E1 superfamily.

    PubMed

    López-Estepa, Miguel; Ardá, Ana; Savko, Martin; Round, Adam; Shepard, William E; Bruix, Marta; Coll, Miquel; Fernández, Francisco J; Jiménez-Barbero, Jesús; Vega, M Cristina

    2015-01-01

    Cyclic N6-threonylcarbamoyladenosine ('cyclic t6A', ct(6)A) is a non-thiolated hypermodification found in transfer RNAs (tRNAs) in bacteria, protists, fungi and plants. In bacteria and yeast cells ct(6)A has been shown to enhance translation fidelity and efficiency of ANN codons by improving the faithful discrimination of aminoacylated tRNAs by the ribosome. To further the understanding of ct(6)A biology we have determined the high-resolution crystal structures of CsdL/TcdA in complex with AMP and ATP, an E1-like activating enzyme from Escherichia coli, which catalyzes the ATP-dependent dehydration of t6A to form ct(6)A. CsdL/TcdA is a dimer whose structural integrity and dimer interface depend critically on strongly bound K+ and Na+ cations. By using biochemical assays and small-angle X-ray scattering we show that CsdL/TcdA can associate with tRNA with a 1:1 stoichiometry and with the proper position and orientation for the cyclization of t6A. Furthermore, we show by nuclear magnetic resonance that CsdL/TcdA engages in transient interactions with CsdA and CsdE, which, in the latter case, involve catalytically important residues. These short-lived interactions may underpin the precise channeling of sulfur atoms from cysteine to CsdL/TcdA as previously characterized. In summary, the combination of structural, biophysical and biochemical methods applied to CsdL/TcdA has afforded a more thorough understanding of how the structure of this E1-like enzyme has been fine tuned to accomplish ct(6)A synthesis on tRNAs while providing support for the notion that CsdA and CsdE are able to functionally interact with CsdL/TcdA.

  6. Characterization and analysis of the Burkholderia pseudomallei BsaN virulence regulon

    PubMed Central

    2014-01-01

    Background Burkholderia pseudomallei is a facultative intracellular pathogen and the causative agent of melioidosis. A conserved type III secretion system (T3SS3) and type VI secretion system (T6SS1) are critical for intracellular survival and growth. The T3SS3 and T6SS1 genes are coordinately and hierarchically regulated by a TetR-type regulator, BspR. A central transcriptional regulator of the BspR regulatory cascade, BsaN, activates a subset of T3SS3 and T6SS1 loci. Results To elucidate the scope of the BsaN regulon, we used RNAseq analysis to compare the transcriptomes of wild-type B. pseudomallei KHW and a bsaN deletion mutant. The 60 genes positively-regulated by BsaN include those that we had previously identified in addition to a polyketide biosynthesis locus and genes involved in amino acid biosynthesis. BsaN was also found to repress the transcription of 51 genes including flagellar motility loci and those encoding components of the T3SS3 apparatus. Using a promoter-lacZ fusion assay in E. coli, we show that BsaN together with the chaperone BicA directly control the expression of the T3SS3 translocon, effector and associated regulatory genes that are organized into at least five operons (BPSS1516-BPSS1552). Using a mutagenesis approach, a consensus regulatory motif in the promoter regions of BsaN-regulated genes was shown to be essential for transcriptional activation. Conclusions BsaN/BicA functions as a central regulator of key virulence clusters in B. pseudomallei within a more extensive network of genetic regulation. We propose that BsaN/BicA controls a gene expression program that facilitates the adaption and intracellular survival of the pathogen within eukaryotic hosts. PMID:25085508

  7. Photonic heat conduction in Josephson-coupled Bardeen-Cooper-Schrieffer superconductors

    NASA Astrophysics Data System (ADS)

    Bosisio, R.; Solinas, P.; Braggio, A.; Giazotto, F.

    2016-04-01

    We investigate the photon-mediated heat flow between two Josephson-coupled Bardeen-Cooper-Schrieffer (BCS) superconductors. We demonstrate that in standard low temperature experiments involving temperature-biased superconducting quantum interference devices (SQUIDs), this radiative contribution is negligible if compared to the direct galvanic one, but it largely exceeds the heat exchanged between electrons and the lattice phonons. The corresponding thermal conductance is found to be several orders of magnitude smaller, for real experiments setup parameters, than the universal quantum of thermal conductance, κ0(T ) =π kB2T /6 ℏ .

  8. A novel Robertsonian translocation event leads to transfer of a stem rust resistance gene (Sr52) effective against race Ug99 from Dasypyrum villosum into bread wheat.

    PubMed

    Qi, L L; Pumphrey, M O; Friebe, Bernd; Zhang, P; Qian, C; Bowden, R L; Rouse, M N; Jin, Y; Gill, B S

    2011-06-01

    Stem rust (Puccinia graminis f. sp. tritici Eriks. & E. Henn.) (the causal agent of wheat stem rust) race Ug99 (also designated TTKSK) and its derivatives have defeated several important stem rust resistance genes widely used in wheat (Triticum aestivum L.) production, rendering much of the worldwide wheat acreage susceptible. In order to identify new resistance sources, a large collection of wheat relatives and genetic stocks maintained at the Wheat Genetic and Genomic Resources Center was screened. The results revealed that most accessions of the diploid relative Dasypyrum villosum (L.) Candargy were highly resistant. The screening of a set of wheat-D. villosum chromosome addition lines revealed that the wheat-D. villosum disomic addition line DA6V#3 was moderately resistant to race Ug99. The objective of the present study was to produce and characterize compensating wheat-D. villosum whole arm Robertsonian translocations (RobTs) involving chromosomes 6D of wheat and 6V#3 of D. villosum through the mechanism of centric breakage-fusion. Seven 6V#3-specific EST-STS markers were developed for screening F(2) progeny derived from plants double-monosomic for chromosomes 6D and 6V#3. Surprisingly, although 6D was the target chromosome, all recovered RobTs involved chromosome 6A implying a novel mechanism for the origin of RobTs. Homozygous translocations (T6AS·6V#3L and T6AL·6V#3S) with good plant vigor and full fertility were selected from F(3) families. A stem rust resistance gene was mapped to the long arm 6V#3L in T6AS·6V#3L and was designated as Sr52. Sr52 is temperature-sensitive and is most effective at 16°C, partially effective at 24°C, and ineffective at 28°C. The T6AS·6V#3L stock is a new source of resistance to Ug99, is cytogenetically stable, and may be useful in wheat improvement.

  9. Theoretical study of the OH addition to the endocyclic and exocyclic double bonds of the d-limonene

    NASA Astrophysics Data System (ADS)

    Ramírez-Ramírez, Víctor M.; Nebot-Gil, Ignacio

    2005-06-01

    The initial step of the d-limonene + OH gas-phase reaction mechanism was investigated by means of ab initio calculations. We have considered eight different possibilities for the OH addition, corresponding to the two C-C double bonds, the two C atoms of each double bond, and the syn or anti orientation, with respect to the isopropenyl group (endocyclic attack) or the ring cycle (exocyclic attack). Activation energies calculated at the QCISD(T)/6-31G(d)//UMP2/6-31G(d) level, show that there are preferred orientations for the OH addition under atmospheric conditions of temperature and pressure.

  10. Engineering of the Stellarator of Costa Rica: SCR-1

    NASA Astrophysics Data System (ADS)

    Mora, J.; Vargas, V. I.; Otarola, C.; Piedra, C.; Jimenez, W.; Esquivel, L.; Esquivel, R.; Sanchez, K.; Gonzalez, J.; Asenjo, J.; Fonseca, L.

    2015-03-01

    This Paper aims at briefly describing the challenge of the design and construction of the Stellarator of Costa Rica 1 (SCR-1) [1]. The SCR-1 is a small modular Stellarator for magnetic confinement of plasma (Ro=0.238 m, =0.059 m, Ro/a>4.0, expected plasma volume ≈ 0.016 m3, 10 mm thickness 6061-T6 aluminum vacuum vessel) developed by the Plasma Laboratory for Fusion Energy and Applications of the Instituto Tecnológico de Costa Rica (ITCR).

  11. Detection of Photosynthetic Performance of Stipa bungeana Seedlings under Climatic Change using Chlorophyll Fluorescence Imaging

    PubMed Central

    Song, Xiliang; Zhou, Guangsheng; Xu, Zhenzhu; Lv, Xiaomin; Wang, Yuhui

    2016-01-01

    In this study, the impact of future climate change on photosynthetic efficiency as well as energy partitioning in the Stipa bungeana was investigated by using chlorophyll fluorescence imaging (CFI) technique. Two thermal regimes (room temperature, T0: 23.0/17.0°C; High temperature, T6: 29.0/23.0°C) and three water conditions (Control, W0; Water deficit, W−30; excess precipitation, W+30) were set up in artificial control chambers. The results showed that excess precipitation had no significant effect on chlorophyll fluorescence parameters, while water deficit decreased the maximal quantum yield of photosystem II (PSII) photochemistry for the dark-adapted state (Fv/Fm) by 16.7%, with no large change in maximal quantum yield of PSII photochemistry for the light-adapted state (FV′/FM′) and coefficient of the photochemical quenching (qP) at T0 condition. Under T6 condition, high temperature offset the negative effect of water deficit on Fv/Fm and enhanced the positive effect of excess precipitation on Fv/Fm, Fv′/Fm′, and qP, the values of which all increased. This indicates that the temperature higher by 6°C will be beneficial to the photosynthetic performance of S. bungeana. Spatial changes of photosynthetic performance were monitored in three areas of interest (AOIs) located on the bottom, middle and upper position of leaf. Chlorophyll fluorescence images (Fv/Fm, actual quantum yield of PSII photochemistry for the light-adapted state (ΦPSII), quantum yield of non-regulated energy dissipation for the light-adapted state (ΦNO) at T0 condition, and ΦPSII at T6 condition) showed a large spatial variation, with greater value of ΦNO and lower values of Fv/Fm and ΦPSII in the upper position of leaves. Moreover, there was a closer relationship between ΦPSII and ΦNO, suggesting that the energy dissipation by non-regulated quenching mechanisms played a dominant role in the yield of PSII photochemistry. It was also found that, among all measured fluorescence

  12. Accelerated universes from type IIA compactifications

    NASA Astrophysics Data System (ADS)

    Blåbäck, Johan; Danielsson, Ulf; Dibitetto, Giuseppe

    2014-03-01

    We study slow-roll accelerating cosmologies arising from geometric compactifications of type IIA string theory on T6/(Bbb Z2 × Bbb Z2). With the aid of a genetic algorithm, we are able to find quasi-de Sitter backgrounds with both slow-roll parameters of order 0.1. Furthermore, we study their evolution by numerically solving the corresponding time-dependent equations of motion, and we show that they actually display a few e-folds of accelerated expansion. Finally, we comment on their perturbative reliability.

  13. Recurrent chondromyxoid fibroma of the thoracic spine 30 years after primary excision: case report and review of the literature.

    PubMed

    Kikuchi, F; Dorfman, H D; Kane, P B

    2001-10-01

    We report a case of late recurrence of chondromyxoid fibroma (CMF) arising in a thoracic vertebra in an 11-year-old male. This was treated by curettage, and 30 years later, the patient noticed shoulder pain and leg weakness. A recurrent mass appeared at the same site in the spinous process of T6. The histologic features of the recurrent tumor were similar to those of the primary lesion. A total of 38 cases of CMF of the vertebra have been reported. Only 3 of 38 previously reported vertebral CMF recurred. Tumors recurred 2 years after operation in 2 cases, and 7 years after operation in 1 case. PMID:12574851

  14. Sorafenib Tosylate and Chemotherapy in Treating Older Patients With Acute Myeloid Leukemia

    ClinicalTrials.gov

    2016-07-05

    Acute Myeloid Leukemia (Megakaryoblastic) With t(1;22)(p13;q13); RBM15-MKL1; Acute Myeloid Leukemia With a Variant RARA Translocation; Acute Myeloid Leukemia With Inv(3)(q21q26.2) or t(3;3)(q21;q26.2); RPN1-EVI1; Acute Myeloid Leukemia With t(6;9)(p23;q34); DEK-NUP214; Acute Myeloid Leukemia With t(9;11)(p22;q23); MLLT3-MLL; Acute Myeloid Leukemia With Variant MLL Translocations; Untreated Adult Acute Myeloid Leukemia

  15. Fatigue and fracture behavior of aluminum-lithium alloys at ambient and cryogenic temperatures

    SciTech Connect

    Xu, Y.B.; Zhang, Y.; Wang, Z.G.; Hu, Z.Q.

    1995-07-15

    The fatigue strength of Al-Li-Cu-Mg-Zr alloys (T6) increases as the test temperature decreases, especially in the long life regime. The fatigue strength is much improved at both room and low temperatures when more zirconium is added. However, the alloy with higher Zr exhibits less-temperatures dependence of the fatigue strength, particularly in the short life regime. Change in fatigue behavior between room and low temperatures are associated with changes is the fracture mode which can be affected significantly by the addition of zirconium.

  16. Performance characterization of fiber Bragg grating thermal response in space vacuum thermal environment

    NASA Astrophysics Data System (ADS)

    Jiang, Junfeng; Song, Luyao; Liu, Tiegen; Zhang, Jingchuan; Liu, Kun; Wang, Shuang; Yin, Jinde; Zhao, Peng; Xie, Jihui; Wu, Fan; Zhang, Xuezhi

    2013-12-01

    We investigated the fiber Bragg grating (FBG) thermal response in space vacuum thermal environment. The FBGs were packaged with 6061-T6 aluminum. The liquid nitrogen immersion experiment results show that its wavelength shift standard deviation is 0.76 pm for 217 h. The combination effect of vacuum and cryogenic temperature was studied by thermal cycling process in space environment simulator. The FBG sensors show accuracy better than 2% full scale, and the hysteresis errors are below 1%. It proves that these metal packaged FBG sensors can survive and meet the requirement of space measurement.

  17. Investigation of Weibull statistics in fracture analysis of cast aluminum

    NASA Technical Reports Server (NTRS)

    Holland, Frederic A., Jr.; Zaretsky, Erwin V.

    1989-01-01

    The fracture strengths of two large batches of A357-T6 cast aluminum coupon specimens were compared by using two-parameter Weibull analysis. The minimum number of these specimens necessary to find the fracture strength of the material was determined. The applicability of three-parameter Weibull analysis was also investigated. A design methodology based on the combination of elementary stress analysis and Weibull statistical analysis is advanced and applied to the design of a spherical pressure vessel shell. The results from this design methodology are compared with results from the applicable ASME pressure vessel code.

  18. Repair of the (6-4) photoproduct by DNA photolyase requires two photons.

    PubMed

    Yamamoto, Junpei; Martin, Ryan; Iwai, Shigenori; Plaza, Pascal; Brettel, Klaus

    2013-07-15

    It takes two (photons) to tango: Single-turnover flash experiments showed that the flavoenzyme (6-4) photolyase uses a successive two-photon mechanism to repair the UV-induced T(6-4)T lesion in DNA (see picture). The intermediate (X) formed by the first photoreaction is likely to be the oxetane-bridged dimer T(ox)T. The enzyme could stabilize the normally short-lived T(ox)T, allowing repair to be completed by the second photoreaction. PMID:23761226

  19. The Corrosion Protection of Metals by Ion Vapor Deposited Aluminum

    NASA Technical Reports Server (NTRS)

    Danford, M. D.

    1993-01-01

    A study of the corrosion protection of substrate metals by ion vapor deposited aluminum (IVD Al) coats has been carried out. Corrosion protection by both anodized and unanodized IVD Al coats has been investigated. Base metals included in the study were 2219-T87 Al, 7075-T6 Al, Titanium-6 Al-4 Vanadium (Ti-6Al-4V), 4130 steel, D6AC steel, and 4340 steel. Results reveal that the anodized IVD Al coats provide excellent corrosion protection, but good protection is also achieved by IVD Al coats that have not been anodized.

  20. Electron beam weld development on a Filter Pack Assembly. Final report

    SciTech Connect

    Dereskiewicz, J.P.

    1994-06-01

    A continuous electron beam welding procedure was developed to replace the manual gas tungsten arc welding procedure on the Filter Pack Assembly. A statistical study was used to evaluate the feasibility of electron beam welding 6061-T6 aluminum covers to A356 cast weldments throughout the joint tolerance range specified on product drawings. Peak temperature exposures were not high enough to degrade the heat sensitive electrical components inside the cast weldment. Actual weldments with alodine coating on the weld joint area were successfully cleaned using a nonmetallic fiberglass brush cleaning method.