Adhesion of epoxy primer to hydrotalcite conversion coated AA2024

NASA Astrophysics Data System (ADS)

Leggat, Robert Benton, III

Hydrotalcite-based (HT) conversion coatings are being developed as an environmentally benign alternative to chromate conversion coatings (CCC). Accelerated exposure tests were conducted on epoxy primed, HT-modified AA2024 to gauge service performance. HT-based conversion coatings did not perform as well as the CCC when used with an epoxy primer. The current HT chemistries are optimized for stand-alone corrosion protection, however additional research into the primer/HT interactions is necessary before they can be implemented within a coating scheme. The relative contribution of mechanical and physico-chemical interactions in controlling adhesion has been investigated in this study. Practical adhesion tests were used to assess the dry and wet bond strength of epoxy primer on HT coatings using the pull-off tensile strength (POTS) as the figure of merit. The practical adhesion of HT coated samples generally fell between that observed for the CCC and bare AA2024. Laboratory testing was done to assess the physical and chemical properties of HT coatings. Contact angle measurements were performed using powders representative of different HT chemistries to evaluate the dispersive and acid-base character of the surface. The wet POTS correlated with the electrodynamic (dipole + dispersive) parameter of the surface tension. The HT surfaces were found to be predominantly basic. Given the basicity of epoxy, these results indicate that increasing the acidic character of HT coatings may increase the adhesion performance. This was supported by electrokinetic measurements in which the dry POTS was found to increase with decreasing conversion coating iso-electric point. The correlations with the dry and wet state adhesion are interpreted as indicating that dry state adhesion is optimized by minimizing unfavorable polar interactions between the basic epoxy and HT interfaces. Wet state adhesion, where polar interactions are disrupted, is dictated by non-polar bonding. FTIR

Effect of wear parameters on dry sliding behavior of Fly Ash/SiC particles reinforced AA 2024 hybrid composites

NASA Astrophysics Data System (ADS)

Bhaskar Kurapati, Vijaya; Kommineni, Ravindra

2017-09-01

In the present work AA 2024 alloy reinforced with mixtures of SiC and Fly Ash (FA) particles of 70 µm (5, 10 and 15 wt. %) are fabricated using the stir casting method. Both reinforcements are added in equal weight proportions. The wear test specimens are prepared from both the alloy and composite castings in the dimensions of Ф 4 mm and 30 mm lengths by the wire cut EDM process. The dry sliding wear properties of the prepared composites at room temperature are estimated by pin-on-disc wear testing equipment. The wear characteristics of the composites are studied by conducting the dry sliding wear test over loads of 0.5 Kgf, 1.0 Kgf, 1.5 Kgf, a track diameter of 60 mm and sliding times of 15 min, 30 min, 45min. The experimental results shows that the wear decreases with an increase in the weight percentage of FA and SiC particles in the matrix. Additionally wear increases with an increase in load and sliding time. Further, it is found that the wear resistance of the AA2024-Hybrid composites is higher than that of the AA2024 matrix.

Sol-gel hybrid films based on organosilane and montmorillonite for corrosion inhibition of AA2024.

PubMed

Dalmoro, V; dos Santos, J H Z; Armelin, E; Alemán, C; Azambuja, D S

2014-07-15

The present work reports the production of films on AA2024-T3 composed of vinyltrimethoxysilane (VTMS)/tetraethylorthosilicate (TEOS) with incorporation of montmorillonite (sodium montmorillonite and montmorillonite modified with quaternary ammonium salt, abbreviated Na and 30B, respectively), generated by the sol-gel process. According to FT-IR analyses the incorporation of montmorillonite does not affect silica network. Electrochemical characterization was performed by electrochemical impedance spectroscopy measurement in 0.05 mol L(-1) NaCl solution. Results indicate that montmorillonite incorporation improves the corrosion protection compared to the non-modified system. Scanning electron microscopy micrographs reveal that high concentrations of montmorillonite provide agglomerations on the metallic surface, which is in detriment of the anticorrosive performance. The VTMS/TEOS/30B films with the lowest concentration (22 mg L(-1)) of embedded clay provide the highest corrosion protection. Copyright © 2014 Elsevier Inc. All rights reserved.

A computational study of low-head direct chill slab casting of aluminum alloy AA2024

NASA Astrophysics Data System (ADS)

Hasan, Mainul; Begum, Latifa

2016-04-01

The steady state casting of an industrial-sized AA2024 slab has been modeled for a vertical low-head direct chill caster. The previously verified 3-D CFD code is used to investigate the solidification phenomena of the said long-range alloy by varying the pouring temperature, casting speed and the metal-mold contact heat transfer coefficient from 654 to 702 °C, 60-180 mm/min, and 1.0-4.0 kW/(m2 K), respectively. The important predicted results are presented and thoroughly discussed.

Organosilane self-assembled layers (SAMs) and hybrid silicate magnesium-rich primers for the corrosion protection of aluminum alloy 2024 T3

NASA Astrophysics Data System (ADS)

Wang, Duhua

Although current chromate coatings function very well in corrosion protection for aircraft alloys, such as aluminum alloy 2024 T3, the U.S. Environmental Protection Agency is planning to totally ban the use of chromates as coating materials in the next decade or so because of their extremely toxic effect. For this purpose, both self-assembled layers and silicate magnesium-rich primers were tested to provide the corrosion protection for aluminum alloy. The long-term goal of this research is to develop a coating system to replace the current chromate coating for aircraft corrosion protection. Aluminum alloy 2024 T3 substrates were modified with self-assembled monolayer or multilayer thin films from different alkylsilane compounds. Mono-functional silanes, such as octadecyltrichlorosilane (C18SiCl3), can form a mixed hydrophobic monolayer or multilayer thin film on the aluminum oxide surface to provide a barrier to water and other electrolytes, so the corrosion resistance of the SAMs modified surface was increased significantly. On the other hand, the bi-functional silane self-assembly could attach the aluminum surface through the silicon headgroup while using its functional tailgroup to chemically bond the polymer coating, thus improving the adhesion between the aluminum substrate and coating substantially, and seems to contribute more to corrosion protection of aluminum substrate. Organosilanes were also combined with tetraethyl orthosilicate (TEOS) in propel ratios to form a sol-gel binder to make silicate magnesium-rich primers. Analogue to the inorganic zinc-rich coatings, the silicate magnesium-rich primers also showed excellent adhesion and solvent resistance. The sacrificial magnesium pigments and the chemically inert silicate binder both contribute to the anti-corrosion properties. Future studies will be focused on the formula optimization for better toughness, chemical resistance and anticorrosion performance.

Auger electron spectroscopy and Rutherford backscattering studies of copper in 2024-T3 aluminum following electrochemical anodization in phosphoric acid

NASA Astrophysics Data System (ADS)

Solomon, J. S.

1981-05-01

The effects of the electrochemical anodization of dioxidized 2024-T3 aluminum on copper were characterized by Auger electron spectroscopy and Rutherford backscattering. Anodization was performed in phosphoric acid at constant potential. Data is presented which shows that constant potential anodization of 2024-T3 is more efficient than aluminum in terms of oxide growth rates for short anodization times. However the maximum anodic oxide thickness achievable on the alloy is less than the pure metal. Copper is shown to be enriched at the oxide metal interface because of its diffusion from the bulk during anodization. The presence of copper at the oxide-metal interface is shown to affect oxide morphology.

A Bottom-Up Optimization Approach for Friction Stir Welding Parameters of Dissimilar AA2024-T351 and AA7075-T651 Alloys

NASA Astrophysics Data System (ADS)

Anil Kumar, K. S.; Murigendrappa, S. M.; Kumar, Hemantha

2017-07-01

In the present study, optimum friction stir weld parameters such as plunge depth, tool rotation speed and traverse speed for butt weld of dissimilar aluminum alloy plates, typically 2024-T351 and 7075-T651, are investigated using a bottom-up approach. In the approach, optimum FSW parameters are achieved by varying any one parameter for every trial while remaining parameters are kept constant. The specimens are extracted from the friction stir-welded plates for studying the tensile, hardness and microstructure properties. Optimum friction stir weld individual parameters are selected based on the highest ultimate tensile strength of the friction stir-welded butt joint specimens produced by varying in each case one parameter and keeping the other two constant. The microstructure samples were investigated for presence of defects, grain refinement at the weld nugget (WN), bonding between the two materials and interface of WN, TMAZ (thermomechanically affected zone) of both advancing and retreating sides of the dissimilar joints using optical microscopy and scanning electron microscopy analyses. In the experimental investigations, the optimum FSW parameters such as plunge depth, 6.2 mm, rotation speed, 650 rpm and traverse speed of 150 mm/min result in ultimate tensile strength, 435 MPa, yield strength, 290 MPa, weld joint efficiency, 92% and maximum elongation, 13%. The microstructure of optimized sample in the WN region revealed alternate lamellae material flow pattern with better metallurgical properties, defect free and very fine equiaxed grain size of about 3-5 µm.

Experimental and Numerical Study of Needle Peening Effects in Aluminium Alloy 2024-T3 Sheets

NASA Astrophysics Data System (ADS)

Mendez Romero, Julio Alberto

Peening of metallic components is an effective treatment used in aerospace and automotive applications to improve fatigue properties or to blend and repair localized damage. This process is typically carried out using metallic airborne media, called shot. However, different processes make use of different media, such is the case of hard, pneumatically powered needles of needle peening equipment. In order to obtain a better understanding of the effects of needle peening in the same context as shot peening, this research work had as an objective to study in detail the behavior of the needle peening equipment in order to characterize the process, design an experimental campaign to measure the effects of needle peening on AA2024-T3 and to develop and validate a Finite Element (FE) model capable of replicating the results of needle peening. The needle peening equipment prototype, called SPIKERRTM, was developed by Shockform Aeronautique Inc. The equipment was characterized by utilizing high-speed camera recording in order to study its behaviour by varying the operating pressure. The obtained collection of images was ran through a newly developed digital image algorithm, so as to quantify the needles' velocity and frequency. The impact velocity and impact frequency were determined for different equipment operating parameters. It was concluded that both the average impact velocity and the impact frequency increase as the pressure becomes larger. Behaviour anomalies among the different needles, such as frequency and velocity variations, were brought to light; these conclusions could be of interest to the manufacturer. Ideally, all of the needles should behave as similarly as possible so as to produce a more uniform process. The response to needle peening of AA2024-T3 in 1.6 mm thick sheet form was studied by needle peening test specimens with dimensions of an Almen strips using the SPIKERRTM. AA2024-T3 was selected since it was extensively studied at Ecole Polytechnique de

Corrosion behavior of ultrafine-grained AA2024 aluminum alloy produced by cryorolling

NASA Astrophysics Data System (ADS)

Laxman Mani Kanta, P.; Srivastava, V. C.; Venkateswarlu, K.; Paswan, Sharma; Mahato, B.; Das, Goutam; Sivaprasad, K.; Krishna, K. Gopala

2017-11-01

The objectives of this study were to produce ultrafine-grained (UFG) AA2024 aluminum alloy by cryorolling followed by aging and to evaluate its corrosion behavior. Solutionized samples were cryorolled to 85% reduction in thickness. Subsequent aging resulted in a UFG structure with finer precipitates of Al2CuMg in the cryorolled alloy. The (1) solutionized and (2) solutionized and cryorolled samples were uniformly aged at 160°C/24 h and were designated as CGPA and CRPA, respectively; these samples were subsequently subjected to corrosion studies. Potentiodynamic polarization studies in 3.5wt% NaCl solution indicated an increase in corrosion potential and a decrease in corrosion current density for CRPA compared to CGPA. In the case of CRPA, electrochemical impedance spectroscopic studies indicated the presence of two complex passive oxide layers with a higher charge transfer resistance and lower mass loss during intergranular corrosion tests. The improved corrosion resistance of CRPA was mainly attributed to its UFG structure, uniform distribution of fine precipitates, and absence of coarse grain-boundary precipitation and associated precipitate-free zones as compared with the CGPA alloy.

Quasi-Static Tensile Stress-Strain Curves. 1, 2024-T3510 Aluminum Alloy

DTIC Science & Technology

1976-02-01

herein were conducted as part of the Core Materials Program of the Solid Mechanics Branch of the Terminal Ballistics Laboratory. The objective of this...describing the results of the Core Materials Program, covers quasi-static terVsile tests of 2024-T3510 aluminum E’lloy. The results include Young’s...11.31 4 580.6 9.94 TABLE II MATERIAL PROPERTIES OF 2024-T3510 ALUMINUM ALLOYa Results of Results of Results of Tensileb Compres ion Sonic Testing

Corrosion of aluminum alloy 2024 by microorganisms isolated from aircraft fuel tanks.

PubMed

McNamara, Christopher J; Perry, Thomas D; Leard, Ryan; Bearce, Ktisten; Dante, James; Mitchell, Ralph

2005-01-01

Microorganisms frequently contaminate jet fuel and cause corrosion of fuel tank metals. In the past, jet fuel contaminants included a diverse group of bacteria and fungi. The most common contaminant was the fungus Hormoconis resinae. However, the jet fuel community has been altered by changes in the composition of the fuel and is now dominated by bacterial contaminants. The purpose of this research was to determine the composition of the microbial community found in fuel tanks containing jet propellant-8 (JP-8) and to determine the potential of this community to cause corrosion of aluminum alloy 2024 (AA2024). Isolates cultured from fuel tanks containing JP-8 were closely related to the genus Bacillus and the fungi Aureobasidium and Penicillium. Biocidal activity of the fuel system icing inhibitor diethylene glycol monomethyl ether is the most likely cause of the prevalence of endospore forming bacteria. Electrochemical impedance spectroscopy and metallographic analysis of AA2024 exposed to the fuel tank environment indicated that the isolates caused corrosion of AA2024. Despite the limited taxonomic diversity of microorganisms recovered from jet fuel, the community has the potential to corrode fuel tanks.

Proof test and fatigue crack growth modeling on 2024-T3 aluminum alloy

NASA Technical Reports Server (NTRS)

Newman, J. C., Jr.; Poe, C. C., Jr.; Dawicke, D. S.

1990-01-01

Pressure proof testing of aircraft fuselage structures has been suggested as a means of screening critical crack sizes and of extending their useful life. The objective of this paper is to study the proof-test concept and to model the crack-growth process on a ductile material. Simulated proof and operational fatigue life tests have been conducted on cracked panels made of 2024-T3 aluminum alloy sheet material. A fatigue crack-closure model was modified to simulate the proof test and operational fatigue cycling. Using crack-growth rate and resistance-curve data, the model was able to predict crack growth during and after the proof load. These tests and analyses indicate that the proof test increases fatigue life; but the beneficial life, after a 1.33 or 1.5 proof, was less than a few hundred cycles.

Influence of Crack-Tip Configurations on the Fracture Response of 0.04-Inch Thick 2024-T3 Aluminum Alloy Sheet

NASA Technical Reports Server (NTRS)

Johnston, William M.; Newman, James C. (Technical Monitor)

2002-01-01

A series of fracture tests were conducted on Middle-crack tension M(T) and compact tension C(T) specimens to determine the effects of specimen type, specimen width, notch tip sharpness and buckling on the fracture behavior of cracked thin sheet (0.04 inch thick) 2024-T3 aluminum alloy material. A series of M(T) specimens were tested with three notch tip configurations: (1) a fatigue pre-cracked notch, (2) a 0.010-inch-diameter wire electrical discharge machined (EDM) notch, and (3) a EDM notch sharpened with a razor blade. The test procedures are discussed and the experimental results for failure stress, load vs. crack extension and the material stress-strain response are reported.

Short fatigue crack behavior in notched 2024-T3 aluminum specimens

NASA Technical Reports Server (NTRS)

Lee, J. J.; Sharpe, W. N., Jr.

1986-01-01

Single-edge, semi-circular notched specimens of Al 2024-T3, 2.3 mm thick, were cyclicly loaded at R-ratios of 0.5, 0.0, -1.0, and -2.0. The notch roots were periodically inspected using a replica technique which duplicates the bore surface. The replicas were examined under an optical microscope to determine the initiation of very short cracks and to monitor the growth of short cracks ranging in length from a few tens of microns to the specimen thickness. In addition to short crack growth measurements, the crack opening displacement (COD) was measured for surface cracks as short as 0.035 mm and for through-thickness cracks using the Interferometric Strain/Displacement Gage (ISDG), a laser-based optical technique. The growth rates of short cracks were faster than the long crack growth rates for R-ratios of -1.0 and -2.0. No significant difference between short and long crack growth rates was observed for R = 0.0. Short cracks had slower growth rates than long cracks for R = 0.5. The crack opening stresses measured for short cracks were smaller than those predicted for large cracks, with little difference appearing for positive R-ratios and large differences noted for negative R-ratios.

On-line Eclipse Resources from the U.S. Naval Observatory: Planning Ahead for April 2024

NASA Astrophysics Data System (ADS)

Fredericks, Amy C.; Bartlett, J. L.; Bell, S.; Stapleton, J. C.

2014-01-01

On 8 April 2024, “…night from mid-day…” (Archilochus, 648 BCE) will appear to fortunate observers along a narrow band, approximately 115 mi (185 km) wide, that crosses fifteen states from Texas to Maine. In response to growing interest in the two total solar eclipses that will sweep the continental United States in the next 11 years, the U.S. Naval Observatory has developed an on-line resource center with direct links to 2024-specific services: the 2024 April 8 Total Solar Eclipse page (http://aa.usno.navy.mil/data/docs/Eclipse2024.php). The Solar Eclipse Computer (http://aa.usno.navy.mil/data/docs/SolarEclipses.php) calculates tables of local circumstances for events visible throughout the world. A similar service is available for lunar eclipses, Lunar Eclipse Computer (http://aa.usno.navy.mil/data/docs/LunarEclipse.php). The USNO Eclipse Portal (http://astro.ukho.gov.uk/eclbin/query_usno.cgi) provides diagrams and animations showing the global circumstances for events visible throughout the world and local circumstances for events visible at selected locations. The Web site, which includes both solar and lunar eclipses, is a joint effort with Her Majesty’s Nautical Almanac Office. The Eclipses of the Sun and Moon page (http://aa.usno.navy.mil/data/docs/UpcomingEclipses.php) links to electronic copies of the visibility maps from The Astronomical Almanac. The Eclipse Reference List (http://aa.usno.navy.mil/faq/docs/eclipse_ref.php) is a representative survey of the available literature for those interested in delving into these phenomena, either technically or historically. As exciting as the 2024 total solar eclipse, another spectacular event will precede it; a total solar eclipse will cross a different swath of the continent on August 21, 2017. The U.S. Naval Observatory has a resource center for that event as well (http://aa.usno.navy.mil/data/docs/Eclipse2017.php) . If your plans for 2024 are not yet made, visit the 2024 April 8 Total Solar Eclipse

Periodic Overload and Transport Spectrum Fatigue Crack Growth Tests of Ti62222STA and Al2024T3 Sheet

NASA Technical Reports Server (NTRS)

Phillips, Edward P.

1999-01-01

Variable amplitude loading crack growth tests have been conducted to provide data that can be used to evaluate crack growth prediction codes. Tests with periodic overloads or overloads followed by underloads were conducted on titanium alloy Ti-6Al-2Sn-2Zr-2Mo-2Cr solution treated and aged (Ti62222STA) material at room temperature and at 350 F. Spectrum fatigue crack growth tests were conducted on two materials (Ti62222STA and aluminum alloy 2024-T3) using two transport lower-wing test spectra at two temperatures (room temperature and 350 F (Ti only)). Test lives (growth from an initial crack half-length of 0.15 in. to failure) were recorded in all tests and the crack length against cycles (or flights) data were recorded in many of the tests. The following observations were made regarding the test results: (1) in tests of the Ti62222STA material, the tests at 350 F had longer lives than those at room temperature, (2) in tests to the MiniTwist spectrum, the Al2024T3 material showed much greater crack growth retardations due to the highest stresses in the spectrum than did the Ti62222STA material, and (3) comparisons of material crack growth performances on an "equal weight" basis were spectrum dependent.

Fatigue crack growth in 2024-T3 aluminum under tensile and transverse shear stresses

NASA Technical Reports Server (NTRS)

Viz, Mark J.; Zehnder, Alan T.

1994-01-01

The influence of transverse shear stresses on the fatigue crack growth rate in thin 2024-T3 aluminum alloy sheets is investigated experimentally. The tests are performed on double-edge cracked sheets in cyclic tensile and torsional loading. This loading generates crack tip stress intensity factors in the same ratio as the values computed for a crack lying along a lap joint in a pressurized aircraft fuselage. The relevant fracture mechanics of cracks in thin plates along with the details of the geometrically nonlinear finite element analyses used for the test specimen calibration are developed and discussed. Preliminary fatigue crack growth data correlated using the fully coupled stress intensity factor calibration are presented and compared with fatigue crack growth data from pure delta K(sub I)fatigue tests.

Overload and Underload Effects on the Fatigue Crack Growth Behavior of the 2024-T3 Aluminum Alloy

NASA Technical Reports Server (NTRS)

Dawicke, David S.

1997-01-01

Fatigue crack growth tests were conducted on 0.09 inch thick, 3.0 inch wide middle-crack tension specimens cut from sheets of 2024-T3 aluminum alloy. The tests were conducted using a load sequence that consisted of a single block of 2,500 cycles of constant amplitude loading followed by an overload/underload combination. The largest fatigue crack growth life occurred for the tests with the overload stress equal to 2 times the constant amplitude stress and the underload stress equal to the constant amplitude minimum stress. For the tests with compressive underloads, the fatigue crack growth life decreased with increasing compressive underload stress.

Molecular Determinants for Substrate Interactions with the Glycine Transporter GlyT2.

PubMed

Carland, Jane E; Thomas, Michael; Mostyn, Shannon N; Subramanian, Nandhitha; O'Mara, Megan L; Ryan, Renae M; Vandenberg, Robert J

2018-03-21

Transporters in the SLC6 family play key roles in regulating neurotransmission and are the targets for a wide range of therapeutics. Important insights into the transport mechanisms and the specificity of drug interactions of SLC6 transporters have been obtained from the crystal structures of a bacterial homologue of the family, LeuT Aa , and more recently the Drosophila dopamine transporter and the human serotonin transporter. However, there is disputed evidence that the bacterial leucine transporter, LeuT Aa , contains two substrate binding sites that work cooperatively in the mechanism of transport, with the binding of a second substrate being required for the release of the substrate from the primary site. An alternate proposal is that there may be low affinity binding sites that serve to direct the flow of substrates to the primary site. We have used a combination of molecular dynamics simulations of substrate interactions with a homology model of GlyT2, together with radiolabeled amino acid uptake assays and electrophysiological analysis of wild-type and mutant transporters, to provide evidence that substrate selectivity of GlyT2 is determined entirely by the primary substrate binding site and, furthermore, if a secondary site exists then it is a low affinity nonselective amino acid binding site.

Effect of the Cold-Sprayed Aluminum Coating-Substrate Interface Morphology on Bond Strength for Aircraft Repair Application

NASA Astrophysics Data System (ADS)

Blochet, Quentin; Delloro, Francesco; N'Guyen, Franck; Jeulin, Dominique; Borit, François; Jeandin, Michel

2017-04-01

This article is dealing with the effects of surface preparation of the substrate on aluminum cold-sprayed coating bond strength. Different sets of AA2024-T3 specimens have been coated with pure Al 1050 feedstock powder, using a conventional cold spray coating technique. The sets were grit-blasted (GB) before coating. The study focuses on substrate surface topography evolution before coating and coating-substrate interface morphology after coating. To study coating adhesion by LASAT® technique for each set, specimens with and without preceding GB treatment were tested in load-controlled conditions. Then, several techniques were used to evaluate the effects of substrate surface treatment on the final coating mechanical properties. Irregularities induced by the GB treatment modify significantly the interface morphology. Results showed that particle anchoring was improved dramatically by the presence of craters. The substrate surface was characterized by numerous anchors. Numerical simulation results exhibited the increasing deformation of particle onto the grit-blasted surface. In addition, results showed a strong relationship between the coating-substrate bond strength on the deposited material and surface preparation.

Microstructure and corrosion resistance of a fluorosilane modified silane-graphene film on 2024 aluminum alloy

NASA Astrophysics Data System (ADS)

Dun, Yuchao; Zhao, Xuhui; Tang, Yuming; Dino, Sahib; Zuo, Yu

2018-04-01

Heptadecafluorodecyl trimethoxysilane (FAS-17) was incorporated into γ-(2,3-epoxypropoxy) propyltrimethoxysilane/graphene (GPTMS/rGO) by adding pre-hydrolyzed FAS-17 solution in GPTMS solution, and a hybrid silane-graphene film (FG/rGO) was prepared on 2024 aluminum alloy surface. The FG/rGO film showed better thermal shock resistance, good adhesion force and high micro-hardness, compared with GPTMS/rGO film. In neutral 3.5 wt% NaCl solution, the corrosion current density for 2024 AA sample with FG/rGO film was 3.40 × 10-3 μA/cm2, which is about one fifth of that for the sample with GPTMS/rGO film. In acidic and alkaline NaCl solutions, the FG/rGO film also showed obviously better corrosion resistance than GPTMS/rGO film. EIS results confirm that the FG/rGO film showed longer performance than GPTMS/rGO film for 2024 AA in NaCl solution. The hydrophobic FAS-17 increased water contact angle of the film surface from 68° to 113°, and changed the stacking structure of graphene in the film. The higher crosslink degree and less interfaces promoted the barrier property of FG/rGO film against aggressive ions and prolonged the performance time in NaCl solution.

Mechanical Properties of Shock Treated Aluminium Alloy Al 2024-T4

NASA Astrophysics Data System (ADS)

Joshi, K. D.; Mukhopadhyay, A. K.; Dey, A.; Rav, Amit S.; Biswas, S.; Gupta, Satish C.

2012-07-01

Plate impact experiment has been carried out on Al 2024-T4 alloy using single stage gas gun. The dynamic yield strength and spall strength of Al 2024-T4 sample has been determined to be 0.35 GPa and 1.43 GPa, respectively, from free surface velocity history measured using VISAR. The sample recovered after unloading from peak shock pressure of 4.4 GPa along with an unshocked sample is analyzed for mechanical properties using nano-indentation and scanning electron microscopy (SEM). The nano-indentation measurements reveal that the hardness and Young's modulus for unshocked sample remains unchanged as a function of load (equivalently depth), however, the same for shocked sample decreases monotonically with increase of load up to ~40 mN and on further increase of load it remains unchanged, suggesting the (i) increase in hardness of shock loaded sample; (ii) the increase in hardness is limited to certain depth, which in our case is 845.12 ± 43.16 nm.

Evaluation of dip and spray coating techniques in corrosion inhibition of AA2024 alloy using a silicon/zirconium sol-gel film as coating

NASA Astrophysics Data System (ADS)

Garcia, R. B. R.; Silva, F. S.; Kawachi, E. Y.

2017-02-01

For corrosion protection of aluminum alloy AA2024 -T3 a silicon/zirconium films were obtained via sol-gel process, prepared from tetraethoxysilane and zirconium acetate, in acid medium with a 5 wt% of nonionic surfactant in order to replace the pre-treatment based on chromium conversion coatings. A homogeneous film was obtained and deposited, at low viscosity condition of the sol (˜10cP), by dip and spray coating techniques. The films morphology was evaluated by Scanning Electron Microscopy (SEM), and to know more about the used deposition methodology, the deposited mass and the film thickness were measured. The corrosion protection efficiency of deposited films was evaluated by potentiodynamic polarization. The film deposition by both dip and spray coatings were effective for the deposition of a homogeneous film layer, and the results showed the thickness is directly related with the deposited mass, and the film deposited by spray technique presented the lower value. Potentiodynamic polarization indicated that the film deposited by spray coating apparently has a better inert ceramic film due the polarization resistance increased around 57% against 27 and 14% of dip coating samples (4 and 1 layer, respectively).

Electrochemical Properties of Organosilane Self Assembled Monolayers on Aluminum 2024

NASA Technical Reports Server (NTRS)

Hintze, Paul E.; Calle, Luz Marina

2004-01-01

Self assembled monolayers are commonly used to modify surfaces. Within the last 15 years, self assembled monolayers have been investigated as a way to protect from corrosion[1,2] or biofouling.[3] In this study, self assembled monolayers of decitriethoxysilane (C10H21Si(OC2H5)3) and octadecyltriethoxysilane (C18H37Si(OC2H5)3) were formed on aluminum 2024-T3. The modified surfaces and bare Al 2024 were characterized by dynamic water contact angle measurements, x-ray photoelectron spectroscopy (XIPS) and infrared spectroscopy. Electrochemical impedance spectroscopy (EIS) in 0.5 M NaCl was used to characterize the monolayers and evaluate their corrosion protection properties. The advancing water contact angle and infrared measurements show that the mono layers form a surface where the hydrocarbon chains are packed and oriented away from the surface, consistent with what is found in similar systems. The contact angle hysteresis measured in these systems is relatively large, perhaps indicating that the hydrocarbon chains are not as well packed as monolayers formed on other substrates. The results of the EIS measurements were modeled using a Randle's circuit modified by changing the capacitor to a constant phase element. The constant phase element values were found to characterize the monolayer. The capacitance of the monolayer modified surface starts lower than the bare Al 2024, but approaches values similar to the bare Al 2024 within 24 hours as the monolayer is degraded. The n values found for bare Al 2024 quickly approach the value of a true capacitor and are greater than 0.9 within hours after the start of exposure. For the monolayer modified structure, n can stay lower than 0.9 for a longer period of time. In fact, n for the monolayer modified surfaces is different from the bare surface even after the capacitance values have converged. This indicates that the deviation from ideal capacitance is the most sensitive indicator of the presence of the monolayer.

Laser shocking of 2024 and 7075 aluminum alloys

NASA Technical Reports Server (NTRS)

Clauer, A. H.; Fairand, B. P.; Slater, J. E.

1977-01-01

The effect of laser generated stress waves on the microstructure, hardness, strength and stress corrosion resistance of 2024 and 7075 aluminum alloys was investigated. Pulsed CO2 and neodymium-glass lasers were used to determine the effect of wavelength and pulse duration on pressure generation and material property changes. No changes in material properties were observed with CO2 laser. The strength and hardness of 2024-T351 and the strength of 7075-T73 aluminum alloys were substantially improved by the stress wave environments generated with the neodymium-glass laser. The mechanical properties of 2024-T851 and 7075-T651 were unchanged by the laser treatment. The correlation of the laser shock data with published results of flyer plate experiments demonstrated that a threshold pressure needed to be exceeded before strengthening and hardening could occur. Peak pressures generated by the pulsed laser source were less than 7.0 GPa which was below the threshold pressure required to change the mechanical properties of 2024-T851 and 7075-T651. Corrosion studies indicated that laser shocking increased the resistance to local attack in 2024-T351 and 7075-T651.

Stress Ratio Effects on Crack Opening Loads and Crack Growth Rates in Aluminum Alloy 2024

NASA Technical Reports Server (NTRS)

Riddell, William T.; Piascik, Robert S.

1998-01-01

The effects of stress ratio (R) and crack opening behavior on fatigue crack growth rates (da/dN) for aluminum alloy (AA) 2024-T3 were investigated using constant-delta K testing, closure measurements, and fractography. Fatigue crack growth rates were obtained for a range of delta K and stress ratios. Results show that constant delta K fatigue crack growth for R ranging from near 0 to 1 is divided into three regions. In Region 1, at low R, da/dN increases with increasing R. In Region 2, at intermediate R, fatigue crack growth rates are relatively independent of R. In Region 3, at high R, further increases in da/dN are observed with increasing R.

Practicing for 2023 and 2024: What the AAS Solar Eclipse Task Force Learned from the "Great American Eclipse" of 2017

NASA Astrophysics Data System (ADS)

Fienberg, R. T.; Speck, A. K.; Habbal, S. R.

2017-12-01

More than three years ahead of the "Great American Eclipse" of August 2017, the American Astronomical Society formed the AAS Solar Eclipse Task Force to function as a think tank, coordinating body, and communication gateway to the vast resources available about the 2017 eclipse and solar eclipses more generally. The task force included professional and amateur astronomers, formal and informal educators, and science journalists; many had experienced total solar eclipses before, and others would experience their first totality in August 2017. The AAS task force secured funding from the AAS Council, the National Science Foundation, and NASA. These resources were used mainly for three purposes: (1) to build a website that contains basic information about solar eclipses, safe viewing practices, and eclipse imaging and video, along with resources for educators and the media and a searchable map of eclipse-related events and activities, with links to other authoritative websites with more detailed information; (2) to solicit, receive, evaluate, and fund proposals for mini-grants to support eclipse-related education and public outreach to underrepresented groups both inside and outside the path of totality; and (3) to organize a series of multidisciplinary workshops across the country to prepare communities for the eclipse and to facilitate collaborations between astronomers, meteorologists, school administrators, and transporation and emergency-management professionals. Most importantly, the AAS Solar Eclipse Task Force focused on developing and disseminating appropriate eclipse safety information. The AAS and NASA jointly developed safety messaging that won the endorsement of the American Academies of Opthalmology and Optometry. In the weeks immediately preceding the eclipse, it became clear that the marketplace was being flooded by counterfeit eclipse glasses and solar viewers, leading to a last minute change in our communication strategy. In this talk, we'll review the

Testing of stiffening ribs formed by incremental forming in thin-walled aircraft structures made of 2024-T3 ALCLAD aluminium alloy

NASA Astrophysics Data System (ADS)

Kubit, Andrzej; Wydrzynski, Dawid; Bucior, Magdalena; Krasowski, Bogdan

2018-05-01

This paper presents the results of experimental tests on the fabrication of longitudinal stiffening ribs in 2024-T3 ALCLAD aluminum alloy sheet, which is widely used in the aircraft structures. The problem presented in this paper concerns the concept of rib-stiffening of the structure of aircraft skin. The ribs are intended to stiffen integral thin-walled structure. Different shapes and different parameters of the forming process were studied. The rib-stiffened samples of various depths of the ribs were tested experimentally in the buckling test.

Dynamic Response and Microstructure Evolution of AA2219-T4 and AA2219-T6 Aluminum Alloys

NASA Astrophysics Data System (ADS)

Olasumboye, A.; Owolabi, G.; Odeshi, A.; Zeytinci, A.; Yilmaz, N.

2018-02-01

In this study, the dynamic deformation behavior of AA2219 aluminum alloy was investigated in two different temper conditions: T4 and T6, with a view to determining the effect of heat treatment on the microstructure and flow behavior of the material under high strain rates. Split Hopkinson pressure bar experiment was used in determining the dynamic response of the alloy while a digital image correlation system was employed in visualizing and tracking the surface deformation of the specimens. Optical microscopy and scanning electron microscopy were used to assess the microstructure of the material after following standard metallographic specimen preparation techniques. The results obtained showed heterogeneous deformation of the alloy in the two temper conditions. It was observed that the dynamic mechanical behavior of each sample preparation was dependent on its strength properties due to aging type, which in turn controls the metamorphosis of the strengthening precipitates and the initial microstructure. At the maximum strain rate of 3500 s-1, transformed bands leading to crack nucleation was observed in the AA2219-T4 aluminum alloy while AA2219-T6 had fractured at the same strain rate. The modes of crack formation and growth in the two alloys were found to be similar: nucleation, growth and coalescence of voids. However, shear band bifurcation phenomenon was observed only in the AA2219-T6 alloy.

Dynamic Response and Microstructure Evolution of AA2219-T4 and AA2219-T6 Aluminum Alloys

NASA Astrophysics Data System (ADS)

Olasumboye, A.; Owolabi, G.; Odeshi, A.; Zeytinci, A.; Yilmaz, N.

2018-06-01

In this study, the dynamic deformation behavior of AA2219 aluminum alloy was investigated in two different temper conditions: T4 and T6, with a view to determining the effect of heat treatment on the microstructure and flow behavior of the material under high strain rates. Split Hopkinson pressure bar experiment was used in determining the dynamic response of the alloy while a digital image correlation system was employed in visualizing and tracking the surface deformation of the specimens. Optical microscopy and scanning electron microscopy were used to assess the microstructure of the material after following standard metallographic specimen preparation techniques. The results obtained showed heterogeneous deformation of the alloy in the two temper conditions. It was observed that the dynamic mechanical behavior of each sample preparation was dependent on its strength properties due to aging type, which in turn controls the metamorphosis of the strengthening precipitates and the initial microstructure. At the maximum strain rate of 3500 s-1, transformed bands leading to crack nucleation was observed in the AA2219-T4 aluminum alloy while AA2219-T6 had fractured at the same strain rate. The modes of crack formation and growth in the two alloys were found to be similar: nucleation, growth and coalescence of voids. However, shear band bifurcation phenomenon was observed only in the AA2219-T6 alloy.

Effect of Multipath Laser Shock Processing on Microhardness, Surface Roughness, and Wear Resistance of 2024-T3 Al Alloy

PubMed Central

Kadhim, Abdulhadi; Salim, Evan T.; Fayadh, Saeed M.; Al-Amiery, Ahmed A.; Kadhum, Abdul Amir H.; Mohamad, Abu Bakar

2014-01-01

Laser shock processing (LSP) is an innovative surface treatment technique with high peak power, short pulse, and cold hardening for strengthening metal materials. LSP is based on the application of a high intensity pulsed laser beam (I > 1 GW/cm2; t < 50 ns) at the interface between the metallic target and the surrounding medium (a transparent confining material, normally water) forcing a sudden vaporization of the metallic surface into a high temperature and density plasma that immediately develops inducing a shock wave propagating into the material. The shock wave induces plastic deformation and a residual stress distribution in the target material. In this paper we study the increase of microhardness and surface roughness with the increase of laser pulse energy in 2024-T3 Al alloy. The influence of the thickness of the confining layer (water) on microhardness and surface roughness is also studied. In addition, the effect of LSP treatment with best conditions on wear behaviors of the alloy was investigated. PMID:24737973

Effect of multipath laser shock processing on microhardness, surface roughness, and wear resistance of 2024-T3 Al alloy.

PubMed

Kadhim, Abdulhadi; Salim, Evan T; Fayadh, Saeed M; Al-Amiery, Ahmed A; Kadhum, Abdul Amir H; Mohamad, Abu Bakar

2014-01-01

Laser shock processing (LSP) is an innovative surface treatment technique with high peak power, short pulse, and cold hardening for strengthening metal materials. LSP is based on the application of a high intensity pulsed laser beam (I > 1 GW/cm(2); t < 50 ns) at the interface between the metallic target and the surrounding medium (a transparent confining material, normally water) forcing a sudden vaporization of the metallic surface into a high temperature and density plasma that immediately develops inducing a shock wave propagating into the material. The shock wave induces plastic deformation and a residual stress distribution in the target material. In this paper we study the increase of microhardness and surface roughness with the increase of laser pulse energy in 2024-T3 Al alloy. The influence of the thickness of the confining layer (water) on microhardness and surface roughness is also studied. In addition, the effect of LSP treatment with best conditions on wear behaviors of the alloy was investigated.

Fracture mechanics data for 2024-T861 and 2124-T851 aluminum

NASA Technical Reports Server (NTRS)

Pionke, L. J.; Linback, R. K.

1974-01-01

The fracture toughness and fatigue flaw growth characteristics of 2024-T861 and 2124-T851 aluminum were evaluated under plane stress conditions. Center cracked tension specimens were employed to evaluate these properties under a number of different test conditions which included variations in specimen thickness, specimen orientation, test environment, and initial flaw size. The effect of buckling was also investigated for all tests of thin gage specimens, and the effect of frequency and stress ratio was evaluated for the cyclic tests. Fracture toughness test results were analyzed and presented in terms of fracture resistance curves; fatigue flaw growth data was analyzed using empirical rate models. The results of the study indicate that both fracture toughness and resistance to fatigue crack growth improve with increasing temperature and decreasing thickness. The presence of buckling during testing of thin gage panels was found to degrade the resistance to fatigue flaw growth only at elevated temperatures.

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

The influence of surface friction on the AA2024 microstructure

NASA Astrophysics Data System (ADS)

Eliseev, A. A.; Kolubaev, E. A.; Fortuna, S. V.

2017-12-01

This work is devoted to the study of the effect of sliding at velocities close to those achieved during friction stir welding or friction drilling on the microstructural evolution of 2024 aluminum alloy. The distribution of both solid solution grains and intermetallic precipitates is analyzed. No layers of recrystallized grains depleted by precipitates, which is a common finding in FSW or friction drilling, are found below the worn surface independently of the sliding velocity. A small precipitate content and size changes alone are observed.

19 CFR 202.4 - Public hearing.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 19 Customs Duties 3 2010-04-01 2010-04-01 false Public hearing. 202.4 Section 202.4 Customs Duties UNITED STATES INTERNATIONAL TRADE COMMISSION NONADJUDICATIVE INVESTIGATIONS INVESTIGATIONS OF COSTS OF PRODUCTION § 202.4 Public hearing. A public hearing will be held in connection with each full investigation...

19 CFR 202.4 - Public hearing.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 19 Customs Duties 3 2012-04-01 2012-04-01 false Public hearing. 202.4 Section 202.4 Customs Duties UNITED STATES INTERNATIONAL TRADE COMMISSION NONADJUDICATIVE INVESTIGATIONS INVESTIGATIONS OF COSTS OF PRODUCTION § 202.4 Public hearing. A public hearing will be held in connection with each full investigation...

19 CFR 202.4 - Public hearing.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 19 Customs Duties 3 2014-04-01 2014-04-01 false Public hearing. 202.4 Section 202.4 Customs Duties UNITED STATES INTERNATIONAL TRADE COMMISSION NONADJUDICATIVE INVESTIGATIONS INVESTIGATIONS OF COSTS OF PRODUCTION § 202.4 Public hearing. A public hearing will be held in connection with each full investigation...

19 CFR 202.4 - Public hearing.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 19 Customs Duties 3 2013-04-01 2013-04-01 false Public hearing. 202.4 Section 202.4 Customs Duties UNITED STATES INTERNATIONAL TRADE COMMISSION NONADJUDICATIVE INVESTIGATIONS INVESTIGATIONS OF COSTS OF PRODUCTION § 202.4 Public hearing. A public hearing will be held in connection with each full investigation...

19 CFR 202.4 - Public hearing.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 19 Customs Duties 3 2011-04-01 2011-04-01 false Public hearing. 202.4 Section 202.4 Customs Duties UNITED STATES INTERNATIONAL TRADE COMMISSION NONADJUDICATIVE INVESTIGATIONS INVESTIGATIONS OF COSTS OF PRODUCTION § 202.4 Public hearing. A public hearing will be held in connection with each full investigation...

Review and study of physics driven pitting corrosion modeling in 2024-T3 aluminum alloys

NASA Astrophysics Data System (ADS)

Yu, Lingyu; Jata, Kumar V.

2015-04-01

Material degradation due to corrosion and corrosion fatigue has been recognized to significantly affect the airworthiness of civilian and military aircraft, especially for the current fleet of airplanes that have served beyond their initial design life. The ability to predict the corrosion damage development in aircraft components and structures, therefore, is of great importance in managing timely maintenance for the aging aircraft vehicles and in assisting the design of new ones. The assessment of aircraft corrosion and its influence on fatigue life relies on appropriate quantitative models that can evaluate the initiation of the corrosion as well as the accumulation during the period of operation. Beyond the aircraft regime, corrosion has also affected the maintenance, safety and reliability of other systems such as nuclear power systems, steam and gas turbines, marine structures and so on. In the work presented in this paper, we reviewed and studied several physics based pitting corrosion models that have been reported in the literature. The classic work of particle induced pitting corrosion by Wei and Harlow is reviewed in detail. Two types of modeling, a power law based simplified model and a microstructure based model, are compared for 2024-T3 alloy. Data from literatures are used as model inputs. The paper ends with conclusions and recommendations for future work.

Reinforcement with alumina particles at the interface region of AA6101-T6 and AA1350 alloys during friction stir welding

NASA Astrophysics Data System (ADS)

Ashok Kumar, R.; Thansekhar, M. R.

2018-04-01

This paper deals the combinational effect of friction stir welding and friction stir processing on dissimilar AA6101-T6 and AA1350 aluminium alloys. For that, alumina particles are reinforced at interface region of AA6101-T6 and AA1350 aluminium alloys. Friction Stir Welding and Friction Stir Processing are done simultaneously for various sizes of groove. To analyze the welding quality and surface modifications, mechanical, wear and microstructural tests are carried out. Among these, smallest groove of 0.5 mm width and 1 mm depth reveals highest tensile and bending strengths and largest groove of 2 mm width and 3 mm depth gives maximum hardness and wear resistance. Taguchi technique shows that groove width is most influencing parameter. Developed second order models with interaction predict the responses with minimum error.

Multiscale approach to micro/macro fatigue crack growth in 2024-T3 aluminum panel

NASA Astrophysics Data System (ADS)

Sih, G. C.

2014-01-01

When two contacting solid surfaces are tightly closed and invisible to the naked eye, the discontinuity is said to be microscopic regardless of whether its length is short or long. By this definition, it is not sufficient to distinguish the difference between a micro- and macro-crack by using the length parameter. Microcracks in high strength metal alloys have been known to be several centimeters or longer. Considered in this work is a dual scale fatigue crack growth model where the main crack can be micro or macro but there prevails an inherent microscopic tip region that is damaged depending on the irregularities of the microstructure. This region is referred to as the "micro-tip" and can be simulated by a sharp wedge with different angles in addition to mixed boundary conditions. The combination is sufficient to model microscopic entities in the form of voids, inclusions, precipitations, interfaces, in addition to subgrain imperfections, or cluster of dislocations. This is accomplished by using the method of "singularity representation" such that closed form asymptotic solutions can be obtained for the development of fatigue crack growth rate relations with three parameters. They include: (1) the crack surface tightness σ* represented by σ o/ σ ∞ = 0.3-0.5 for short cracks in region I, and 0.1-0.2 for long cracks in region II, (2) the micro/macro material properties reflected by the shear modulus ratio µ* (=µmicro/µmacro varying between 2 and 5) and (3) the most sensitive parameter d* being the micro-tip characteristic length d* (= d/ d o) whose magnitude decreases in the direction of region I→II. The existing fatigue crack growth data for 2024-T3 and 7075-T6 aluminum sheets are used to reinterpret the two-parameter d a/d N= C(Δ K) n relation where Δ K has now been re-derived for a microcrack with surfaces tightly in contact. The contact force will depend on the mean stress σm or mean stress ratio R as the primary parameter and on the stress

Identification of novel substrates of Shigella T3SA through analysis of its virulence plasmid-encoded secretome

PubMed Central

Pinaud, Laurie; Ferrari, Mariana L.; Friedman, Robin; Jehmlich, Nico; von Bergen, Martin; Phalipon, Armelle; Sansonetti, Philippe J.

2017-01-01

Many human Gram-negative bacterial pathogens express a Type Three Secretion Apparatus (T3SA), including among the most notorious Shigella spp., Salmonella enterica, Yersinia enterocolitica and enteropathogenic Escherichia coli (EPEC). These bacteria express on their surface multiple copies of the T3SA that mediate the delivery into host cells of specific protein substrates critical to pathogenesis. Shigella spp. are Gram-negative bacterial pathogens responsible for human bacillary dysentery. The effector function of several Shigella T3SA substrates has largely been studied but their potential cellular targets are far from having been comprehensively delineated. In addition, it is likely that some T3SA substrates have escaped scrutiny as yet. Indeed, sequencing of the virulence plasmid of Shigella flexneri has revealed numerous open reading frames with unknown functions that could encode additional T3SA substrates. Taking advantage of label-free mass spectrometry detection of proteins secreted by a constitutively secreting strain of S. flexneri, we identified five novel substrates of the T3SA. We further confirmed their secretion through the T3SA and translocation into host cells using β-lactamase assays. The coding sequences of two of these novel T3SA substrates (Orf13 and Orf131a) have a guanine-cytosine content comparable to those of T3SA components and effectors. The three other T3SA substrates identified (Orf48, Orf86 and Orf176) have significant homology with antitoxin moieties of type II Toxin-Antitoxin systems usually implicated in the maintenance of low copy plasmids. While Orf13 and Orf131a might constitute new virulence effectors contributing to S. flexneri pathogenicity, potential roles for the translocation into host cells of antitoxins or antitoxin-like proteins during Shigella infection are discussed. PMID:29073283

Characterization of lap joints laser beam welding of thin AA 2024 sheets with Yb:YAG disk-laser

NASA Astrophysics Data System (ADS)

Caiazzo, Fabrizia; Alfieri, Vittorio; Cardaropoli, Francesco; Sergi, Vincenzo

2012-06-01

Lap joints obtained by overlapping two plates are widely diffused in aerospace industry. Nevertheless, because of natural aging, adhesively bonded and riveted aircraft lap joints may be affected by cracks from rivets, voids or corrosion. Friction stir welding has been proposed as a valid alternative, although large heat affected zones are produced both in the top and the bottom plate due to the pin diameter. Interest has therefore been shown in studying laser lap welding as the laser beam has been proved to be competitive since it allows to concentrate the thermal input and increases productivity and quality. Some challenges arise as a consequence of aluminum low absorptance and high thermal conductivity; furthermore, issues are due to metallurgical challenges such as both micro and macro porosity formation and softening in the fused zone. Welding of AA 2024 thin sheets in a lap joint configuration is discussed in this paper: tests are carried out using a recently developed Trumpf TruDisk 2002 Yb:YAG disk-laser with high beam quality which allows to produce beads with low plates distortion and better penetration. The influence of the processing parameters is discussed considering the fused zone extent and the bead shape. The porosity content as well as the morphological features of the beads have been examined.

Dynamic Multivariate Accelerated Corrosion Test Protocol

DTIC Science & Technology

2014-10-01

atmospheric, accelerated, AA2024-T3, AA6061-T6, AA7075-T3, 1010 steel, AgCl, rare earth conversion coat, magnesium rich primer, polyurethane , Eyring, Monte...morphology and elemental analysis by scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) and electrochemical determinations of...in the FT-IR analysis; degradation of the components of the high performance polyurethane coatings exposed in the UV/ozone chamber were more

42 CFR 423.2024 - Objections to the issues.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 3 2010-10-01 2010-10-01 false Objections to the issues. 423.2024 Section 423.2024 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES..., and Judicial Review § 423.2024 Objections to the issues. (a) If an enrollee objects to the issues...

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. Copyright © 2014 Elsevier B.V. All rights reserved.

Four-point-bend fatigue of AA 2026 aluminum alloys

NASA Astrophysics Data System (ADS)

Li, J. X.; Zhai, T.; Garratt, M. D.; Bray, G. H.

2005-09-01

High-cycle fatigue tests were carried out on a newly developed high-strength AA 2026 Al alloy, which was in the form of extrusion bars with square and rectangular cross sections, using a self-aligning four-point-bend rig at room temperature, 15 Hz, and R = 0.1, in lab air. The fatigue strength of the square and rectangular bars was measured to be 85 and 90 pct of their yield strength, respectively, more than twice that of the predecessor to the 2026 alloy (the AA 2024 Al alloy). Fatigue cracks were found to be always initiated at large Θ' (Al7Cu2(Fe,Mn)) particles and to propagate predominantly in a crystallographic mode in the AA 2026 alloy. The fatigue fractographies of the square and rectangular extrusion bars were found to be markedly different, due to their different grain structures (fibril and layered, respectively). Fracture steps on the crack face were found in both of these extrusion bars. Since the 2026 alloy was purer in terms of Fe and Si content, it contained much less coarse particles than in a 2024 alloy. This partially accounted for the superior fatigue strength of the 2026 alloy.

Uniform Corrosion and General Dissolution of Aluminum Alloys 2024-T3, 6061-T6, and 7075-T6

NASA Astrophysics Data System (ADS)

Huang, I.-Wen

Uniform corrosion and general dissolution of aluminum alloys was not as well-studied in the past, although it was known for causing significant amount of weight loss. This work comprises four chapters to understand uniform corrosion of aluminum alloys 2024-T3, 6061-T6, and 7075-T6. A preliminary weight loss experiment was performed for distinguishing corrosion induced weight loss attributed to uniform corrosion and pitting corrosion. The result suggested that uniform corrosion generated a greater mass loss than pitting corrosion. First, to understand uniform corrosion mechanism and kinetics in different environments, a series of static immersion tests in NaCl solutions were performed to provide quantitative measurement of uniform corrosion. Thereafter, uniform corrosion development as a function of temperature, pH, Cl-, and time was investigated to understand the influence of environmental factors. Faster uniform corrosion rate has been found at lower temperature (20 and 40°C) than at higher temperature (60 and 80°C) due to accelerated corrosion product formation at high temperatures inhibiting corrosion reactions. Electrochemical tests including along with scanning electron microscopy (SEM) were utilized to study the temperature effect. Second, in order to further understand the uniform corrosion influence on pit growth kinetics, a long term exposures for 180 days in both immersion and ASTM-B117 test were performed. Uniform corrosion induced surface recession was found to have limited impact on pit geometry regardless of exposure methods. It was also found that the competition for limited cathodic current from uniform corrosion the primary rate limiting factor for pit growth. Very large pits were found after uniform corrosion growth reached a plateau due to corrosion product coverage. Also, optical microscopy and focused ion beam (FIB) imaging has provided more insights of distinctive pitting geometry and subsurface damages found from immersion samples and B117

Effect of CrO3 Sealing Time on Anodized A12024-T3

NASA Astrophysics Data System (ADS)

Korda, Akhmad A.; Hidayat, R. Z.

2016-08-01

The effect of CrO3 sealing time on anodized aluminum alloy has been investigated. A1 2024-T3 were used as substrate. Anodizing was carried out using chromic acid. CrO3 sealing was conducted in CrO3 solution for 30, 60, 90, 120 and 150 minutes. As comparison, other specimens were also prepared as anodized and boiled water sealing. Thickness of the coating was observed by optical microscope. Anodized and sealing layer was analyzed by X- ray diffraction. The hardness of as anodized, boiled water sealing and CrO3 sealing were compared. The highest hardness is achieved by CrO3 sealed specimen and followed by boiled water sealing and as anodized specimens. The longer the processes of CrO3 sealing the higher layer thickness and therefore the higher hardness of the oxide layer. The best resistance to electrolyte penetration is achieved by the CrO3 sealed specimen followed by boiled water sealed and as anodized specimens. The higher thickness of oxide layer, the higher the resistance against electrolyte penetration.

Tensile Properties of Friction Stir Welded Joints of AA 2024-T6 Alloy at Different Welding Speeds

NASA Astrophysics Data System (ADS)

Avula, Dhananjayulu; Devuri, Venkateswarlu; Cheepu, Muralimohan; Dwivedi, Dheerendra Kumar

2018-03-01

The influence of welding speed on the friction stir welded joint properties of hardness, tensile properties, defects and microstructure characterization are studied in the present study. The friction stir welding was conducted on AA2014-T6 heat treated alloy with 5 mm thickness plate in butt joint configuration. The welding speed was varied from 8 mm/min to 120 mm/min at the fixed travel speed and load conditions. It is observed that the welding speeds at higher rate with wide range can be possible to weld this alloy at higher rates of tool revolution suggesting that the inherent capability of friction stir welding technique for aluminum 2014 alloys. The strength of the joints gradually increases with enhancing of welding speed. The micro structural observations exhibited the formation of equiaxed grains in the stir zone and slightly in the thermo-mechanically affected zone. In addition, the size of the grains decreases with increase in welding speed owing to the presence of low heat input. Hence the hardness of the joints slightly increased in the stir zones over the other zones of the weld nugget. The joint strength initially increases with the welding speed and starts to decreases after reaching to the maximum value. The relationship between the welding conditions and friction stir welded joint properties has been discussed.

Rationally evolving tRNAPyl for efficient incorporation of noncanonical amino acids

PubMed Central

Fan, Chenguang; Xiong, Hai; Reynolds, Noah M.; Söll, Dieter

2015-01-01

Genetic encoding of noncanonical amino acids (ncAAs) into proteins is a powerful approach to study protein functions. Pyrrolysyl-tRNA synthetase (PylRS), a polyspecific aminoacyl-tRNA synthetase in wide use, has facilitated incorporation of a large number of different ncAAs into proteins to date. To make this process more efficient, we rationally evolved tRNAPyl to create tRNAPyl-opt with six nucleotide changes. This improved tRNA was tested as substrate for wild-type PylRS as well as three characterized PylRS variants (Nϵ-acetyllysyl-tRNA synthetase [AcKRS], 3-iodo-phenylalanyl-tRNA synthetase [IFRS], a broad specific PylRS variant [PylRS-AA]) to incorporate ncAAs at UAG codons in super-folder green fluorescence protein (sfGFP). tRNAPyl-opt facilitated a 5-fold increase in AcK incorporation into two positions of sfGFP simultaneously. In addition, AcK incorporation into two target proteins (Escherichia coli malate dehydrogenase and human histone H3) caused homogenous acetylation at multiple lysine residues in high yield. Using tRNAPyl-opt with PylRS and various PylRS variants facilitated efficient incorporation of six other ncAAs into sfGFP. Kinetic analyses revealed that the mutations in tRNAPyl-opt had no significant effect on the catalytic efficiency and substrate binding of PylRS enzymes. Thus tRNAPyl-opt should be an excellent replacement of wild-type tRNAPyl for future ncAA incorporation by PylRS enzymes. PMID:26250114

Coarsening of AA6013-T6 Precipitates During Sheet Warm Forming Applications

NASA Astrophysics Data System (ADS)

Di Ciano, M.; DiCecco, S.; Esmaeili, S.; Wells, M. A.; Worswick, M. J.

2018-03-01

The use of warm forming for AA6xxx-T6 sheet is of interest to improve its formability; however, the effect warm forming may have on the coarsening of precipitates and the mechanical strength of these sheets has not been well studied. In this research, the coarsening behavior of AA6013-T6 precipitates has been explored, in the temperature range of 200-300 °C, and time of 30 s up to 50 h. Additionally, the effect of warm deformation on coarsening behavior was explored using: (1) simulated warm forming tests in a Gleeble thermo-mechanical simulator and (2) bi-axial warm deformation tests. Using a strong obstacle model to describe the yield strength (YS) evolution of the AA6013-T6 material, and a Lifshitz, Slyozov, and Wagner (LSW) particle coarsening law to describe the change in precipitate size with time, the coarsening kinetics were modeled for this alloy. The coarsening kinetics in the range of 220-300 °C followed a trend similar to that previously found for AA6111 for the 180-220 °C range. There was strong evidence that coarsening kinetics were not altered due to warm deformation above 220 °C. For warm forming between 200 and 220 °C, the YS of the AA6013-T6 material increased slightly, which could be attributed to strain hardening during warm deformation. Finally, a non-isothermal coarsening model was used to assess the potential reduction in the YS of AA6013-T6 for practical processing conditions related to auto-body manufacturing. The model calculations showed that 90% of the original AA6013-T6 YS could be maintained, for warm forming temperatures up to 280 °C, if the heating schedule used to get the part to the warm forming temperature was limited to 1 min.

Interfacial and Mechanical Behavior of AA5456 Filling Friction-Stir-Welded Lap Joints Using Similar and Dissimilar Pins

NASA Astrophysics Data System (ADS)

Behmand, Saleh Alaei; Mirsalehi, Seyyed Ehsan; Omidvar, Hamid; Safarkhanian, Mohammad Ali

2016-10-01

In this article, filling friction stir welding (FFSW) of the remaining exit holes of AA5456 alloy friction-stir-welded lap joints was studied. For this purpose, the influences of different rotating speeds, holding times, and pin materials, AA5456 and AA2024, on the metallurgical structure and joint strength were investigated. The observations showed that defect-free lap joints are successfully obtainable by this method using similar and dissimilar consumable pins. The results indicated that the higher rotating speed and holding time adversely affect the weld performance. The best result was achieved for 30 seconds holding time, 500 rpm rotating speed, and AA2024 consumable pin. In this condition, a lap shear strength of 10 pct higher than that of the nonfilled joint, equivalent to about 94 pct of the original defect-free FSW joint, was obtained, whereas the GTAW filled joint showed only approximately 87 pct of the continuous FSW joint strength.

48 CFR 2024.202 - Policy.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 48 Federal Acquisition Regulations System 6 2014-10-01 2014-10-01 false Policy. 2024.202 Section 2024.202 Federal Acquisition Regulations System NUCLEAR REGULATORY COMMISSION SOCIOECONOMIC PROGRAMS PROTECTION OF PRIVACY AND FREEDOM OF INFORMATION Freedom of Information Act 2024.202 Policy. The provisions...

48 CFR 2024.202 - Policy.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 48 Federal Acquisition Regulations System 6 2012-10-01 2012-10-01 false Policy. 2024.202 Section 2024.202 Federal Acquisition Regulations System NUCLEAR REGULATORY COMMISSION SOCIOECONOMIC PROGRAMS PROTECTION OF PRIVACY AND FREEDOM OF INFORMATION Freedom of Information Act 2024.202 Policy. The provisions...

48 CFR 2024.202 - Policy.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 48 Federal Acquisition Regulations System 6 2010-10-01 2010-10-01 true Policy. 2024.202 Section 2024.202 Federal Acquisition Regulations System NUCLEAR REGULATORY COMMISSION SOCIOECONOMIC PROGRAMS PROTECTION OF PRIVACY AND FREEDOM OF INFORMATION Freedom of Information Act 2024.202 Policy. The provisions...

20 CFR 202.4 - Control.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 20 Employees' Benefits 1 2011-04-01 2011-04-01 false Control. 202.4 Section 202.4 Employees' Benefits RAILROAD RETIREMENT BOARD REGULATIONS UNDER THE RAILROAD RETIREMENT ACT EMPLOYERS UNDER THE ACT § 202.4 Control. A company or person is controlled by one or more carriers, whenever there exists in one...

20 CFR 202.4 - Control.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 20 Employees' Benefits 1 2013-04-01 2012-04-01 true Control. 202.4 Section 202.4 Employees' Benefits RAILROAD RETIREMENT BOARD REGULATIONS UNDER THE RAILROAD RETIREMENT ACT EMPLOYERS UNDER THE ACT § 202.4 Control. A company or person is controlled by one or more carriers, whenever there exists in one...

20 CFR 202.4 - Control.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 20 Employees' Benefits 1 2012-04-01 2012-04-01 false Control. 202.4 Section 202.4 Employees' Benefits RAILROAD RETIREMENT BOARD REGULATIONS UNDER THE RAILROAD RETIREMENT ACT EMPLOYERS UNDER THE ACT § 202.4 Control. A company or person is controlled by one or more carriers, whenever there exists in one...

20 CFR 202.4 - Control.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false Control. 202.4 Section 202.4 Employees' Benefits RAILROAD RETIREMENT BOARD REGULATIONS UNDER THE RAILROAD RETIREMENT ACT EMPLOYERS UNDER THE ACT § 202.4 Control. A company or person is controlled by one or more carriers, whenever there exists in one...

20 CFR 202.4 - Control.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 20 Employees' Benefits 1 2014-04-01 2012-04-01 true Control. 202.4 Section 202.4 Employees' Benefits RAILROAD RETIREMENT BOARD REGULATIONS UNDER THE RAILROAD RETIREMENT ACT EMPLOYERS UNDER THE ACT § 202.4 Control. A company or person is controlled by one or more carriers, whenever there exists in one...

Experimental study on joining of AA6063 and AISI 1040 steel

NASA Astrophysics Data System (ADS)

Hynes, N. Rajesh Jesudoss; Raja, S.

2018-05-01

Feasibility of joining of dissimilar metals with different physical, chemical and thermal properties such as AA6063 alloy and AISI 1040 steel is worthwhile study, since it has tremendous applications in all most of all engineering domains. The mechanism of bonding is studied using scanning electron microscopy. Impact strength of AA2024/AISI joints, axial shortening distance, micro hardness distribution and joint strength are determined. Micro hardness profile shows increased hardness value at the joint interface, due to grain refinement.

Activity of vegetative insecticidal proteins Vip3Aa58 and Vip3Aa59 of Bacillus thuringiensis against lepidopteran pests.

PubMed

Baranek, Jakub; Kaznowski, Adam; Konecka, Edyta; Naimov, Samir

2015-09-01

Vegetative insecticidal proteins (Vips) secreted by some isolates of Bacillus thuringiensis show activity against insects and are regarded as insecticides against pests. A number of B. thuringiensis strains harbouring vip3A genes were isolated from different sources and identified by using a PCR based approach. The isolates with the highest insecticidal activity were indicated in screening tests, and their vip genes were cloned and sequenced. The analysis revealed two polymorphic Vip protein forms, which were classified as Vip3Aa58 and Vip3Aa59. After expression of the vip genes, the proteins were isolated and characterized. The activity of both toxins was estimated against economically important lepidopteran pests of woodlands (Dendrolimus pini), orchards (Cydia pomonella) and field crops (Spodoptera exigua). Vip3Aa58 and Vip3Aa59 were highly toxic and their potency surpassed those of many Cry proteins used in commercial bioinsecticides. Vip3Aa59 revealed similar larvicidal activity as Vip3Aa58 against S. exigua and C. pomonella. Despite 98% similarity of amino acid sequences of both proteins, Vip3Aa59 was significantly more active against D. pini. Additionally the effect of proteolytic activation of Vip58Aa and Vip3Aa59 on toxicity of D. pini and S. exigua was studied. Both Vip3Aa proteins did not show any activity against Tenebrio molitor (Coleoptera) larvae. The results suggest that the Vip3Aa58 and Vip3Aa59 toxins might be useful for controlling populations of insect pests of crops and forests. Copyright © 2015 Elsevier Inc. All rights reserved.

Evaluating stress corrosion cracking behaviour of high strength AA7075-T651 aluminium alloy

NASA Astrophysics Data System (ADS)

Prabhuraj, P.; Rajakumar, S.; Lakshminarayanan, A. K.; Balasubramanian, V.

2017-12-01

The objective of the present study is to determine the threshold stress level of stress corrosion cracking (SCC) in AA7075-T651 aluminium alloy by suitable experimentation. The test was carried out using a circumferential notch specimen in a horizontal-type constant load SCC setup in a 3.5 wt.% NaCl solution. The time to failure by SCC was determined at various loading conditions. The threshold stress of AA7075-T651 alloy was found to be 242 MPa in a 3.5 wt.% NaCl solution. The various regions of the fractured surface specimen such as machined notch, SCC region and final overload fracture area were examined using scanning electron microscopy (SEM) in order to identify the SCC mechanism.

A Novel Tenebrio molitor Cadherin Is a Functional Receptor for Bacillus thuringiensis Cry3Aa Toxin*

PubMed Central

Fabrick, Jeff; Oppert, Cris; Lorenzen, Marcé D.; Morris, Kaley; Oppert, Brenda; Jurat-Fuentes, Juan Luis

2009-01-01

Cry toxins produced by the bacterium Bacillus thuringiensis are effective biological insecticides. Cadherin-like proteins have been reported as functional Cry1A toxin receptors in Lepidoptera. Here we present data that demonstrate that a coleopteran cadherin is a functional Cry3Aa toxin receptor. The Cry3Aa receptor cadherin was cloned from Tenebrio molitor larval midgut mRNA, and the predicted protein, TmCad1, has domain structure and a putative toxin binding region similar to those in lepidopteran cadherin B. thuringiensis receptors. A peptide containing the putative toxin binding region from TmCad1 bound specifically to Cry3Aa and promoted the formation of Cry3Aa toxin oligomers, proposed to be mediators of toxicity in lepidopterans. Injection of TmCad1-specific double-stranded RNA into T. molitor larvae resulted in knockdown of the TmCad1 transcript and conferred resistance to Cry3Aa toxicity. These data demonstrate the functional role of TmCad1 as a Cry3Aa receptor in T. molitor and reveal similarities between the mode of action of Cry toxins in Lepidoptera and Coleoptera. PMID:19416969

Rationally evolving tRNAPyl for efficient incorporation of noncanonical amino acids.

PubMed

Fan, Chenguang; Xiong, Hai; Reynolds, Noah M; Söll, Dieter

2015-12-15

Genetic encoding of noncanonical amino acids (ncAAs) into proteins is a powerful approach to study protein functions. Pyrrolysyl-tRNA synthetase (PylRS), a polyspecific aminoacyl-tRNA synthetase in wide use, has facilitated incorporation of a large number of different ncAAs into proteins to date. To make this process more efficient, we rationally evolved tRNA(Pyl) to create tRNA(Pyl-opt) with six nucleotide changes. This improved tRNA was tested as substrate for wild-type PylRS as well as three characterized PylRS variants (N(ϵ)-acetyllysyl-tRNA synthetase [AcKRS], 3-iodo-phenylalanyl-tRNA synthetase [IFRS], a broad specific PylRS variant [PylRS-AA]) to incorporate ncAAs at UAG codons in super-folder green fluorescence protein (sfGFP). tRNA(Pyl-opt) facilitated a 5-fold increase in AcK incorporation into two positions of sfGFP simultaneously. In addition, AcK incorporation into two target proteins (Escherichia coli malate dehydrogenase and human histone H3) caused homogenous acetylation at multiple lysine residues in high yield. Using tRNA(Pyl-opt) with PylRS and various PylRS variants facilitated efficient incorporation of six other ncAAs into sfGFP. Kinetic analyses revealed that the mutations in tRNA(Pyl-opt) had no significant effect on the catalytic efficiency and substrate binding of PylRS enzymes. Thus tRNA(Pyl-opt) should be an excellent replacement of wild-type tRNA(Pyl) for future ncAA incorporation by PylRS enzymes. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

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.

Effect of Segregation of Secondary Phase Particles and "S" Line on Tensile Fracture Behavior of Friction Stir-Welded 2024Al-T351 Joints

NASA Astrophysics Data System (ADS)

Zhang, Z.; Xiao, B. L.; Ma, Z. Y.

2013-09-01

A 5-mm-thick 2024Al-T351 plate was friction stir welded (FSWed) at welding speeds of 100, 200, and 400 mm min-1 with a constant rotation rate of 800 rpm, and the microstructure and tensile fracture behavior of the joints were investigated in detail. FSW resulted in the redistribution of secondary phase particles along the recrystallized grain boundaries at the nugget zone (NZ), forming linear segregation bands consisting of secondary phase particles. The segregation bands, mainly present in the shoulder-driven zone, were believed to result from periodic material flow, with the average band spacing on the longitudinal and horizontal cross sections equal to the tool advancement per revolution. At a low welding speed of 100 mm min-1, in spite of the highest density of segregation bands, the FSWed 2024Al-T351 joint fractured along the low hardness zone (LHZ) of the heat-affected zone because of large hardness gap between NZ and LHZ. Increasing the welding speed to 200 and 400 mm min-1 reduced both the hardness gap between NZ and LHZ and the density of segregation bands. In this case, the segregation bands played a role, resulting in unusual fracture of the joints along the segregation bands. The "S" line originated from the oxide film on the initial butting surfaces and did not affect the fracture behavior of the FSWed 2024Al-T351 joints.

Acoustic emission monitoring of tensile testing of corroded and un-corroded clad aluminum 2024-T3 and characterization of effects of corrosion on AE source events and material tensile properties

NASA Astrophysics Data System (ADS)

Okafor, A. Chukwujekwu; Natarajan, Shridhar

2014-02-01

Corrosion damage affects structural integrity and deteriorates material properties of aluminum alloys in aircraft structures. Acoustic Emission (AE) is an effective nondestructive evaluation (NDE) technique for monitoring such damages and predicting failure in large structures of an aircraft. For successful interpretation of data from AE monitoring, sources of AE and factors affecting it need to be identified. This paper presents results of AE monitoring of tensile testing of corroded and un-corroded clad Aluminum 2024-T3 test specimens, and characterization of the effects of strain-rate and corrosion damage on material tensile properties and AE source events. Effect of corrosion was studied by inducing corrosion in the test specimens by accelerated corrosion testing in a Q-Fog accelerated corrosion chamber for 12 weeks. Eight (8) masked dog-bone shaped specimens were placed in the accelerated corrosion chamber at the beginning of the test. Two (2) dog-bone shaped specimens were removed from the corrosion chamber after exposure time of 3, 6, 9, and 12 weeks respectively, and subjected to tension testing till specimen failure along with AE monitoring, as well as two (2) reference samples not exposed to corrosion. Material tensile properties (yield strength, ultimate tensile strength, toughness, and elongation) obtained from tension test and AE parameters obtained from AE monitoring were analyzed and characterized. AE parameters increase with increase in exposure period of the specimens in the corrosive environment. Aluminum 2024-T3 is an acoustically silent material during tensile deformation without any damage. Acoustic emission events increase with increase of corrosion damage and with increase in strain rate above a certain value. Thus AE is suitable for structural health monitoring of corrosion damage. Ultimate tensile strength, toughness and elongation values decrease with increase of exposure period in corrosion chamber.

19 CFR 10.2024 - Indirect materials.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 19 Customs Duties 1 2014-04-01 2014-04-01 false Indirect materials. 10.2024 Section 10.2024... Agreement Rules of Origin § 10.2024 Indirect materials. An indirect material, as defined in § 10.2013(i), will be considered to be an originating material without regard to where it is produced. Example...

Is the presence of AA amyloidosis associated with impaired coronary flow reserve?

PubMed

Bulut, Mustafa; Keles, Nursen; Caliskan, Zuhal; Kostek, Osman; Aksu, Feyza; Ozdil, Kamil; Akcakoyun, Mustafa; Demircioglu, Kenan; Yilmaz, Yusuf; Kanbay, Mehmet; Caliskan, Mustafa

2016-08-01

Systemic amyloid A protein (AA) amyloidosis may occur as a complication of many chronic inflammatory disorders. Patients receiving inadequate anti-inflammatory and immunosuppressive therapies have an increased risk of developing systemic AA amyloidosis. Inflammation plays a role in all stages and the thrombotic complications of atherosclerosis. In the absence of epicardial coronary stenosis, coronary flow reserve (CFR) reflects coronary microvascular dysfunction. In the present study, we hypothesized that amyloid advanced subclinical inflammation in chronic inflammatory diseases (CID) patients may further affect coronary microcirculation. Thirty-two patients with biopsy-diagnosed renal AA, 73 patients with non-amyloid CID, and a group of healthy volunteers were included in the study. The measurements of coronary flow velocity were performed by a single investigator with expertise in transthoracic Doppler harmonic echocardiography (TTDE). The AA amyloidosis subgroup had significantly lower CFR values than other non-amyloid CID patients and the control individuals (1.8 (1.5-2.1) vs. 2.1 (2.0-2.4) and 3.0 (2.8-3.2), p < 0.001). Multivariate logistic regression analysis indicated that the presence of AA amyloidosis and elevated hs - CRP independently predict impairment of the CFR (p < 0.05). The presence of AA amyloidosis is related to decreased CFR values and the presence of AA amyloidosis and elevated hs - CRP independently predict impairment of the CFR. Therefore, patients with AA amyloidosis may have an increased risk of developing coronary artery diseases. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

The cytochrome P450 2AA gene cluster in zebrafish (Danio rerio): Expression of CYP2AA1 and CYP2AA2 and response to phenobarbital-type inducers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kubota, Akira; Bainy, Afonso C.D.; Departamento de Bioquímica, CCB, Universidade Federal de Santa Catarina, Florianopolis, SC 88040-900

2013-10-01

The cytochrome P450 (CYP) 2 gene family is the largest and most diverse CYP gene family in vertebrates. In zebrafish, we have identified 10 genes in a new subfamily, CYP2AA, which does not show orthology to any human or other mammalian CYP genes. Here we report evolutionary and structural relationships of the 10 CYP2AA genes and expression of the first two genes, CYP2AA1 and CYP2AA2. Parsimony reconstruction of the tandem duplication pattern for the CYP2AA cluster suggests that CYP2AA1, CYP2AA2 and CYP2AA3 likely arose in the earlier duplication events and thus are most diverged in function from the other CYP2AAs.more » On the other hand, CYP2AA8 and CYP2AA9 are genes that arose in the latest duplication event, implying functional similarity between these two CYPs. A molecular model of CYP2AA1 showing the sequence conservation across the CYP2AA cluster reveals that the regions with the highest variability within the cluster map onto CYP2AA1 near the substrate access channels, suggesting differing substrate specificities. Zebrafish CYP2AA1 transcript was expressed predominantly in the intestine, while CYP2AA2 was most highly expressed in the kidney, suggesting differing roles in physiology. In the liver CYP2AA2 expression but not that of CYP2AA1, was increased by 1,4-bis [2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP) and, to a lesser extent, by phenobarbital (PB). In contrast, pregnenolone 16α-carbonitrile (PCN) increased CYP2AA1 expression, but not CYP2AA2 in the liver. The results identify a CYP2 subfamily in zebrafish that includes genes apparently induced by PB-type chemicals and PXR agonists, the first concrete in vivo evidence for a PB-type response in fish. - Highlights: • A tandemly duplicated cluster of ten CYP2AA genes was described in zebrafish. • Parsimony and duplication analyses suggest pathways to CYP2AA diversity. • Homology models reveal amino acid positions possibly related to functional diversity. • The CYP2AA locus does not share

Transcriptome profiling of the intoxication response of Tenebrio molitor larvae to Bacillus thuringiensis Cry3Aa protoxin.

PubMed

Oppert, Brenda; Dowd, Scot E; Bouffard, Pascal; Li, Lewyn; Conesa, Ana; Lorenzen, Marcé D; Toutges, Michelle; Marshall, Jeremy; Huestis, Diana L; Fabrick, Jeff; Oppert, Cris; Jurat-Fuentes, Juan Luis

2012-01-01

Bacillus thuringiensis (Bt) crystal (Cry) proteins are effective against a select number of insect pests, but improvements are needed to increase efficacy and decrease time to mortality for coleopteran pests. To gain insight into the Bt intoxication process in Coleoptera, we performed RNA-Seq on cDNA generated from the guts of Tenebrio molitor larvae that consumed either a control diet or a diet containing Cry3Aa protoxin. Approximately 134,090 and 124,287 sequence reads from the control and Cry3Aa-treated groups were assembled into 1,318 and 1,140 contigs, respectively. Enrichment analyses indicated that functions associated with mitochondrial respiration, signalling, maintenance of cell structure, membrane integrity, protein recycling/synthesis, and glycosyl hydrolases were significantly increased in Cry3Aa-treated larvae, whereas functions associated with many metabolic processes were reduced, especially glycolysis, tricarboxylic acid cycle, and fatty acid synthesis. Microarray analysis was used to evaluate temporal changes in gene expression after 6, 12 or 24 h of Cry3Aa exposure. Overall, microarray analysis indicated that transcripts related to allergens, chitin-binding proteins, glycosyl hydrolases, and tubulins were induced, and those related to immunity and metabolism were repressed in Cry3Aa-intoxicated larvae. The 24 h microarray data validated most of the RNA-Seq data. Of the three intoxication intervals, larvae demonstrated more differential expression of transcripts after 12 h exposure to Cry3Aa. Gene expression examined by three different methods in control vs. Cry3Aa-treated larvae at the 24 h time point indicated that transcripts encoding proteins with chitin-binding domain 3 were the most differentially expressed in Cry3Aa-intoxicated larvae. Overall, the data suggest that T. molitor larvae mount a complex response to Cry3Aa during the initial 24 h of intoxication. Data from this study represent the largest genetic sequence dataset for T. molitor

Transcriptome Profiling of the Intoxication Response of Tenebrio molitor Larvae to Bacillus thuringiensis Cry3Aa Protoxin

PubMed Central

Oppert, Brenda; Dowd, Scot E.; Bouffard, Pascal; Li, Lewyn; Conesa, Ana; Lorenzen, Marcé D.; Toutges, Michelle; Marshall, Jeremy; Huestis, Diana L.; Fabrick, Jeff; Oppert, Cris; Jurat-Fuentes, Juan Luis

2012-01-01

Bacillus thuringiensis (Bt) crystal (Cry) proteins are effective against a select number of insect pests, but improvements are needed to increase efficacy and decrease time to mortality for coleopteran pests. To gain insight into the Bt intoxication process in Coleoptera, we performed RNA-Seq on cDNA generated from the guts of Tenebrio molitor larvae that consumed either a control diet or a diet containing Cry3Aa protoxin. Approximately 134,090 and 124,287 sequence reads from the control and Cry3Aa-treated groups were assembled into 1,318 and 1,140 contigs, respectively. Enrichment analyses indicated that functions associated with mitochondrial respiration, signalling, maintenance of cell structure, membrane integrity, protein recycling/synthesis, and glycosyl hydrolases were significantly increased in Cry3Aa-treated larvae, whereas functions associated with many metabolic processes were reduced, especially glycolysis, tricarboxylic acid cycle, and fatty acid synthesis. Microarray analysis was used to evaluate temporal changes in gene expression after 6, 12 or 24 h of Cry3Aa exposure. Overall, microarray analysis indicated that transcripts related to allergens, chitin-binding proteins, glycosyl hydrolases, and tubulins were induced, and those related to immunity and metabolism were repressed in Cry3Aa-intoxicated larvae. The 24 h microarray data validated most of the RNA-Seq data. Of the three intoxication intervals, larvae demonstrated more differential expression of transcripts after 12 h exposure to Cry3Aa. Gene expression examined by three different methods in control vs. Cry3Aa-treated larvae at the 24 h time point indicated that transcripts encoding proteins with chitin-binding domain 3 were the most differentially expressed in Cry3Aa-intoxicated larvae. Overall, the data suggest that T. molitor larvae mount a complex response to Cry3Aa during the initial 24 h of intoxication. Data from this study represent the largest genetic sequence dataset for T. molitor

Co-expression of AaPMT and AaTRI effectively enhances the yields of tropane alkaloids in Anisodus acutangulus hairy roots

PubMed Central

2011-01-01

Background Tropane alkaloids (TA) including anisodamine, anisodine, hyoscyamine and scopolamine are a group of important anticholinergic drugs with rapidly increasing market demand, so it is significant to improve TA production by biotechnological approaches. Putrescine N-methyltransferase (PMT) was considered as the first rate-limiting upstream enzyme while tropinone reductase I (TRI) was an important branch-controlling enzyme involved in TA biosynthesis. However, there is no report on simultaneous introduction of PMT and TRI genes into any TA-producing plant including Anisodus acutangulus (A. acutangulus), which is a Solanaceous perennial plant that is endemic to China and is an attractive resource plant for production of TA. Results In this study, 21 AaPMT and AaTRI double gene transformed lines (PT lines), 9 AaPMT single gene transformed lines (P lines) and 5 AaTRI single gene transformed lines (T lines) were generated. RT-PCR and real-time fluorescence quantitative analysis results revealed that total AaPMT (AaPMT T) and total AaTRI (AaTRI T) gene transcripts in transgenic PT, P and T lines showed higher expression levels than native AaPMT (AaPMT E) and AaTRI (AaTRI E) gene transcripts. As compared to the control and single gene transformed lines (P or T lines), PT transgenic hairy root lines produced significantly higher levels of TA. The highest yield of TA was detected as 8.104 mg/g dw in line PT18, which was 8.66, 4.04, and 3.11-times higher than those of the control (0.935 mg/g dw), P3 (highest in P lines, 2.004 mg/g dw) and T12 (highest in T lines, 2.604 mg/g dw), respectively. All the tested samples were found to possess strong radical scavenging capacity, which were similar to control. Conclusion In the present study, the co-expression of AaPMT and AaTRI genes in A. acutangulus hairy roots significantly improved the yields of TA and showed higher antioxidant activity than control because of higher total TA content, which is the first report on

Membrane-bound human orphan cytochrome P450 2U1: Sequence singularities, construction of a full 3D model, and substrate docking.

PubMed

Ducassou, Lionel; Dhers, Laura; Jonasson, Gabriella; Pietrancosta, Nicolas; Boucher, Jean-Luc; Mansuy, Daniel; André, François

2017-09-01

Human cytochrome P450 2U1 (CYP2U1) is an orphan CYP that exhibits several distinctive characteristics among the 57 human CYPs with a highly conserved sequence in almost all living organisms. We compared its protein sequence with those of the 57 human CYPs and constructed a 3D structure of a full-length CYP2U1 model bound to a POPC membrane. We also performed docking experiments of arachidonic acid (AA) and N-arachidonoylserotonin (AS) in this model. The protein sequence of CYP2U1 displayed two unique characteristics when compared to those of the human CYPs, the presence of a longer N-terminal region upstream of the putative trans-membrane helix (TMH) containing 8 proline residues, and of an insert of about 20 amino acids containing 5 arginine residues between helices A' and A. Its N-terminal part upstream of TMH involved an additional short terminal helix, in a manner similar to what was reported in the crystal structure of Saccharomyces cerevisiae CYP51. Our model also showed a specific interaction between the charged residues of insert AA' and phosphate groups of lipid polar heads, suggesting a possible role of this insert in substrate recruitment. Docking of AA and AS in this model showed these substrates in channel 2ac, with the terminal alkyl chain of AA or the indole ring of AS close to the heme, in agreement with the reported CYP2U1-catalyzed AA and AS hydroxylation regioselectivities. This model should be useful to find new endogenous or exogenous CYP2U1 substrates and to interpret the regioselectivity of their hydroxylation. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Crack-tip-opening angle measurements and crack tunneling under stable tearing in thin sheet 2024-T3 aluminum alloy

NASA Technical Reports Server (NTRS)

Dawicke, D. S.; Sutton, M. A.

1993-01-01

The stable tearing behavior of thin sheets 2024-T3 aluminum alloy was studied for middle crack tension specimens having initial cracks that were: flat cracks (low fatigue stress) and 45 degrees through-thickness slant cracks (high fatigue stress). The critical crack-tip-opening angle (CTOA) values during stable tearing were measured by two independent methods, optical microscopy and digital image correlation. Results from the two methods agreed well. The CTOA measurements and observations of the fracture surfaces showed that the initial stable tearing behavior of low and high fatigue stress tests is significantly different. The cracks in the low fatigue stress tests underwent a transition from flat-to-slant crack growth, during which the CTOA values were high and significant crack tunneling occurred. After crack growth equal to about the thickness, CTOA reached a constant value of 6 deg and after crack growth equal to about twice the thickness, crack tunneling stabilized. The initial high CTOA values, in the low fatigue crack tests, coincided with large three-dimensional crack front shape changes due to a variation in the through-thickness crack tip constraint. The cracks in the high fatigue stress tests reach the same constant CTOA value after crack growth equal to about the thickness, but produced only a slightly higher CTOA value during initial crack growth. For crack growth on the 45 degree slant, the crack front and local field variables are still highly three-dimensional. However, the constant CTOA values and stable crack front shape may allow the process to be approximated with two-dimensional models.

Identification of Bacillus thuringiensis Cry3Aa toxin domain II loop 1 as the binding site of Tenebrio molitor cadherin repeat CR12.

PubMed

Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Amaro, Itzel; Ortíz, Ernesto; Becerril, Baltazar; Ibarra, Jorge E; Bravo, Alejandra; Soberón, Mario

2015-04-01

Bacillus thuringiensis Cry toxins exert their toxic effect by specific recognition of larval midgut proteins leading to oligomerization of the toxin, membrane insertion and pore formation. The exposed domain II loop regions of Cry toxins have been shown to be involved in receptor binding. Insect cadherins have shown to be functionally involved in toxin binding facilitating toxin oligomerization. Here, we isolated a VHH (VHHA5) antibody by phage display that binds Cry3Aa loop 1 and competed with the binding of Cry3Aa to Tenebrio molitor brush border membranes. VHHA5 also competed with the binding of Cry3Aa to a cadherin fragment (CR12) that was previously shown to be involved in binding and toxicity of Cry3Aa, indicating that Cry3Aa binds CR12 through domain II loop 1. Moreover, we show that a loop 1 mutant, previously characterized to have increased toxicity to T. molitor, displayed a correlative enhanced binding affinity to T. molitor CR12 and to VHHA5. These results show that Cry3Aa domain II loop 1 is a binding site of CR12 T. molitor cadherin. Copyright © 2015 Elsevier Ltd. All rights reserved.

Characterization, Modeling, and Accelerating Emulation of Aircraft Coating Exposure and Degradation

DTIC Science & Technology

2009-09-30

nucleation sites for conjugated polymer electrodeposition on AA 2024-T3. In particular, the role of secondary phase heterogeneities in the nucleation ...work is mainly contained in the MS Thesis of T. Chen, and was presented publicly on October 14, 2010 at the NACE Eastern Area Conference in

Active corrosion protection of AA2024 by sol-gel coatings with corrosion inhibitors =

NASA Astrophysics Data System (ADS)

Yasakau, Kiryl

A industria aeronautica utiliza ligas de aluminio de alta resistencia para o fabrico dos elementos estruturais dos avioes. As ligas usadas possuem excelentes propriedades mecanicas mas apresentam simultaneamente uma grande tendencia para a corrosao. Por esta razao essas ligas necessitam de proteccao anticorrosiva eficaz para poderem ser utilizadas com seguranca. Ate a data, os sistemas anticorrosivos mais eficazes para ligas de aluminio contem cromio hexavalente na sua composicao, sejam pre-tratamentos, camadas de conversao ou pigmentos anticorrosivos. O reconhecimento dos efeitos carcinogenicos do cromio hexavalente levou ao aparecimento de legislacao banindo o uso desta forma de cromio pela industria. Esta decisao trouxe a necessidade de encontrar alternativas ambientalmente inocuas mas igualmente eficazes. O principal objectivo do presente trabalho e o desenvolvimento de pretratamentos anticorrosivos activos para a liga de aluminio 2024, baseados em revestimentos hibridos produzidos pelo metodo sol-gel. Estes revestimentos deverao possuir boa aderencia ao substrato metalico, boas propriedades barreira e capacidade anticorrosiva activa. A proteccao activa pode ser alcancada atraves da incorporacao de inibidores anticorrosivos no pretratamento. O objectivo foi atingido atraves de uma sucessao de etapas. Primeiro investigou-se em detalhe a corrosao localizada (por picada) da liga de aluminio 2024. Os resultados obtidos permitiram uma melhor compreensao da susceptibilidade desta liga a processos de corrosao localizada. Estudaram-se tambem varios possiveis inibidores de corrosao usando tecnicas electroquimicas e microestruturais. Numa segunda etapa desenvolveram-se revestimentos anticorrosivos hibridos organico-inorganico baseados no metodo sol-gel. Compostos derivados de titania e zirconia foram combinados com siloxanos organofuncionais a fim de obter-se boa aderencia entre o revestimento e o substrato metalico assim como boas propriedades barreira. Testes

Investigation of Three Analytical Hypothesis for Determining Material Creep Behavior under Varied Loads, with an Application to 2024-T3 Aluminum-Alloy Sheet in Tension at 400 F

NASA Technical Reports Server (NTRS)

Berkovits, Avraham

1961-01-01

Three existing hypotheses are formulated mathematically to estimate tensile creep strain under varied loads and constant temperature from creep data obtained under constant load and constant temperature. hypotheses investigated include the time-hardening, strain-hardening, and life-fraction rules. Predicted creep behavior is compared with data obtained from tensile creep tests of 2024-T3 aluminum-alloy sheet at 400 F under cyclic-load conditions. creep strain under varied loads is presented on the basis of an equivalent stress, derived from the life-fraction rule, which reduces the varied-load case to a constant-load problem. Creep strain in the region of interest for structural design and rupture times, determined from the hypotheses investigated, are in fair agreement with data in most cases, although calculated values of creep strain are generally greater than the experimental values because creep recovery is neglected in the calculations.

How EF-Tu can contribute to efficient proofreading of aa-tRNA by the ribosome

PubMed Central

Noel, Jeffrey K.; Whitford, Paul C.

2016-01-01

It has long been recognized that the thermodynamics of mRNA–tRNA base pairing is insufficient to explain the high fidelity and efficiency of aminoacyl-tRNA (aa-tRNA) selection by the ribosome. To rationalize this apparent inconsistency, Hopfield proposed that the ribosome may improve accuracy by utilizing a multi-step kinetic proofreading mechanism. While biochemical, structural and single-molecule studies have provided a detailed characterization of aa-tRNA selection, there is a limited understanding of how the physical–chemical properties of the ribosome enable proofreading. To this end, we probe the role of EF-Tu during aa-tRNA accommodation (the proofreading step) through the use of energy landscape principles, molecular dynamics simulations and kinetic models. We find that the steric composition of EF-Tu can reduce the free-energy barrier associated with the first step of accommodation: elbow accommodation. We interpret this effect within an extended kinetic model of accommodation and show how EF-Tu can contribute to efficient and accurate proofreading. PMID:27796304

Semisolid Microstructural Evolution during Partial Remelting of a Bulk Alloy Prepared by Cold Pressing of the Ti-Al-2024Al Powder Mixture

PubMed Central

Qin, Yahong; Chen, Tijun; Wang, Yingjun; Zhang, Xuezheng; Li, Pubo

2016-01-01

A new method, powder thixoforming, has been proposed to fabricate an in situ Al3Tip/2024Al composite. During partial remelting, the microstructural evolution of the bulk alloy prepared by cold pressing of the Ti, Al, 2024Al powder mixture was investigated, and the formation mechanism of the Al3Ti particles produced by the reaction between the Ti powder and the Al alloy melt is also discussed in detail. The results indicate that the microstructural evolution of the 2024 alloy matrix can be divided into three stages: a rapid coarsening of the powder grains; a formation of primary α-Al particles surrounded with a continuous liquid film; and a slight coarsening of the primary α-Al particles. Simultaneously, a reaction layer of Al3Ti can be formed on the Ti powder surface when the bulk is heated for 10 min at 640 °C The thickness (X) of the reaction layer increases with the time according to the parabolic law of X=−0.43t2+4.21t+0.17. The stress generated in the reaction layer due to the volume dilatation can be calculated by using the equation σAl3Ti=−EAl3Ti6(1−υAl3Ti)t2Al3TitTi(1R−1R0). Comparing the obtained data with the results of the drip experiment, the reaction rate for the Ti powder and Al powder mixture is greater than that for the Ti plate and Al alloy mixture, respectively.

12 CFR 202.4 - General rules.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 12 Banks and Banking 2 2010-01-01 2010-01-01 false General rules. 202.4 Section 202.4 Banks and Banking FEDERAL RESERVE SYSTEM BOARD OF GOVERNORS OF THE FEDERAL RESERVE SYSTEM EQUAL CREDIT OPPORTUNITY... languages other than English, provided they are available in English upon request. [Reg. B, 68 FR 13161, Mar...

12 CFR 202.4 - General rules.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 12 Banks and Banking 2 2011-01-01 2011-01-01 false General rules. 202.4 Section 202.4 Banks and Banking FEDERAL RESERVE SYSTEM BOARD OF GOVERNORS OF THE FEDERAL RESERVE SYSTEM EQUAL CREDIT OPPORTUNITY... by §§ 202.5 and 202.13, in a form the applicant may retain. (2) Disclosures in electronic form. The...

Methionine Regulates mTORC1 via the T1R1/T1R3-PLCβ-Ca2+-ERK1/2 Signal Transduction Process in C2C12 Cells.

PubMed

Zhou, Yuanfei; Ren, Jiao; Song, Tongxing; Peng, Jian; Wei, Hongkui

2016-10-11

The mammalian target of rapamycin complex 1 (mTORC1) integrates amino acid (AA) availability to support protein synthesis and cell growth. Taste receptor type 1 member (T1R) is a G protein-coupled receptor that functions as a direct sensor of extracellular AA availability to regulate mTORC1 through Ca 2+ stimulation and extracellular signal-regulated kinases 1 and 2 (ERK1/2) activation. However, the roles of specific AAs in T1R1/T1R3-regulated mTORC1 are poorly defined. In this study, T1R1 and T1R3 subunits were expressed in C2C12 myotubes, and l-AA sensing was accomplished by T1R1/T1R3 to activate mTORC1. In response to l-AAs, such as serine (Ser), arginine (Arg), threonine (Thr), alanine (Ala), methionine (Met), glutamine (Gln), and glycine (Gly), Met induced mTORC1 activation and promoted protein synthesis. Met also regulated mTORC1 via T1R1/T1R3-PLCβ-Ca 2+ -ERK1/2 signal transduction. Results revealed a new role for Met-regulated mTORC1 via an AA receptor. Further studies should be performed to determine the role of T1R1/T1R3 in mediating extracellular AA to regulate mTOR signaling and to reveal its mechanism.

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%.

Akt substrate TBC1D1 regulates GLUT1 expression through the mTOR pathway in 3T3-L1 adipocytes

PubMed Central

Zhou, Qiong L.; Jiang, Zhen Y.; Holik, John; Chawla, Anil; Hagan, G. Nana; Leszyk, John; Czech, Michael P.

2010-01-01

Multiple studies have suggested that the protein kinase Akt/PKB (protein kinase B) is required for insulin-stimulated glucose transport in skeletal muscle and adipose cells. In an attempt to understand links between Akt activation and glucose transport regulation, we applied mass spectrometry-based proteomics and bioinformatics approaches to identify potential Akt substrates containing the phospho-Akt substrate motif RXRXXpS/T. The present study describes the identification of the Rab GAP (GTPase-activating protein)-domain containing protein TBC1D1 [TBC (Tre-2/Bub2/Cdc16) domain family, member 1], which is closely related to TBC1D4 [TBC domain family, member 4, also denoted AS160 (Akt substrate of 160 kDa)], as an Akt substrate that is phosphorylated at Thr590. RNAi (RNA interference)-me-diated silencing of TBC1D1 elevated basal deoxyglucose uptake by approx. 61% in 3T3-L1 mouse embryo adipocytes, while the suppression of TBC1D4 and RapGAP220 under the same conditions had little effect on basal and insulin-stimulated deoxy-glucose uptake. Silencing of TBC1D1 strongly increased expression of the GLUT1 glucose transporter but not GLUT4 in cultured adipocytes, whereas the decrease in TBC1D4 had no effect. Remarkably, loss of TBC1D1 in 3T3-L1 adipocytes activated the mTOR (mammalian target of rapamycin)-p70 S6 protein kinase pathway, and the increase in GLUT1 expression in the cells treated with TBC1D1 siRNA (small interfering RNA) was blocked by the mTOR inhibitor rapamycin. Furthermore, overexpression of the mutant TBC1D1-T590A, lacking the putative Akt/PKB phosphorylation site, inhibited insulin stimulation of p70 S6 kinase phosphorylation at Thr389, a phosphorylation induced by mTOR. Taken together, our data suggest that TBC1D1 may be involved in controlling GLUT1 glucose transporter expression through the mTOR-p70 S6 kinase pathway. PMID:18215134

Improvement on the Fatigue Performance of 2024-T4 Alloy by Synergistic Coating Technology

PubMed Central

Wang, Xi-Shu; Guo, Xing-Wu; Li, Xu-Dong; Ge, Dong-Yun

2014-01-01

In this paper, rotating bending fatigue tests of 2024-T4 Al alloy with different oxide coatings were carried out. Compared to the uncoated and previously reported oxide coatings of aluminum alloys, the fatigue strength is able to be enhanced by using a novel oxide coating with sealing pore technology. These results indicate that the better the coating surface quality is, the more excellent the fatigue performance under rotating bending fatigue loading is. The improvement on the fatigue performance is mainly because the fatigue crack initiation and the early stage of fatigue crack growth at the coating layer can be delayed after PEO coating with pore sealing. Therefore, it is a so-called synergistic coating technology for various uses, including welding thermal cracks and filling micro-pores. The effects of different oxide coatings on surface hardness, compressive residual stress, morphology and fatigue fracture morphology are discussed. A critical compressive residual stress of about 95–100 MPa is proposed. PMID:28788634

Galvanic Couple Current and Potential Distribution between a Mg Electrode and 2024-T351 under Droplets Analyzed by Microelectrode Arrays

DTIC Science & Technology

2015-11-04

cathodically to – 1.5 V vs. SCE in aearated NaCl solution dissolves at a rate as high as 4 mA/cm2. However, the same alloy was shown to avoid significant...MgRP coating could be utilized in place of the Mg electrode. It is well known that the wettability of the surface of aluminum and its oxides is high ...aluminum alloys , such as precipitation age hardened 2024-T351, and has performed well in field studies.1–13 The MgRP is designed to be applied to an

Multi-Objective Optimization of Friction Stir Welding Process Parameters of AA6061-T6 and AA7075-T6 Using a Biogeography Based Optimization Algorithm

PubMed Central

Tamjidy, Mehran; Baharudin, B. T. Hang Tuah; Paslar, Shahla; Matori, Khamirul Amin; Sulaiman, Shamsuddin; Fadaeifard, Firouz

2017-01-01

The development of Friction Stir Welding (FSW) has provided an alternative approach for producing high-quality welds, in a fast and reliable manner. This study focuses on the mechanical properties of the dissimilar friction stir welding of AA6061-T6 and AA7075-T6 aluminum alloys. The FSW process parameters such as tool rotational speed, tool traverse speed, tilt angle, and tool offset influence the mechanical properties of the friction stir welded joints significantly. A mathematical regression model is developed to determine the empirical relationship between the FSW process parameters and mechanical properties, and the results are validated. In order to obtain the optimal values of process parameters that simultaneously optimize the ultimate tensile strength, elongation, and minimum hardness in the heat affected zone (HAZ), a metaheuristic, multi objective algorithm based on biogeography based optimization is proposed. The Pareto optimal frontiers for triple and dual objective functions are obtained and the best optimal solution is selected through using two different decision making techniques, technique for order of preference by similarity to ideal solution (TOPSIS) and Shannon’s entropy. PMID:28772893

Multi-Objective Optimization of Friction Stir Welding Process Parameters of AA6061-T6 and AA7075-T6 Using a Biogeography Based Optimization Algorithm.

PubMed

Tamjidy, Mehran; Baharudin, B T Hang Tuah; Paslar, Shahla; Matori, Khamirul Amin; Sulaiman, Shamsuddin; Fadaeifard, Firouz

2017-05-15

The development of Friction Stir Welding (FSW) has provided an alternative approach for producing high-quality welds, in a fast and reliable manner. This study focuses on the mechanical properties of the dissimilar friction stir welding of AA6061-T6 and AA7075-T6 aluminum alloys. The FSW process parameters such as tool rotational speed, tool traverse speed, tilt angle, and tool offset influence the mechanical properties of the friction stir welded joints significantly. A mathematical regression model is developed to determine the empirical relationship between the FSW process parameters and mechanical properties, and the results are validated. In order to obtain the optimal values of process parameters that simultaneously optimize the ultimate tensile strength, elongation, and minimum hardness in the heat affected zone (HAZ), a metaheuristic, multi objective algorithm based on biogeography based optimization is proposed. The Pareto optimal frontiers for triple and dual objective functions are obtained and the best optimal solution is selected through using two different decision making techniques, technique for order of preference by similarity to ideal solution (TOPSIS) and Shannon's entropy.

Microstructure Evolution during Dissimilar Friction Stir Welding of AA7003-T4 and AA6060-T4.

PubMed

Dong, Jialiang; Zhang, Datong; Zhang, Weiwen; Zhang, Wen; Qiu, Cheng

2018-02-27

In this work, the dissimilar joint of AA7003-T4 and 6060-T4 alloy has been produced by friction stir welding (FSW). The microstructure was examined by optical microscope (OM), electron back scattered diffraction (EBSD), transmission electron microscopy (TEM), and the mechanical properties of the joint were investigated. It is demonstrated that sound dissimilar joint can be produced through FSW. In the nugget; precipitations dissolve into the matrix and η' reprecipitate subsequently; and the elongated aluminum grains are replaced by fine and equiaxed grains due to dynamic recrystallization (DRX). In the heat affected zone (HAZ), coarse β' and η precipitates are formed and the aluminum grains are coarser as compared to the base materials. In the thermo-mechanical affected zone (TMAZ), equiaxed and elongated grains coexist due to incomplete DRX. The ultimate tensile strength of the dissimilar joint is 159.2 MPa and its elongation is 10.4%. The weak area exists in the HAZ of 6060 alloy, which is placed in the retreating side during FSW. The correlations between the microstucture and mechanical properties of the dissimilar joint are discussed.

Finite Element Simulation of Temperature and Strain Distribution during Friction Stir Welding of AA2024 Aluminum Alloy

NASA Astrophysics Data System (ADS)

Jain, Rahul; Pal, Surjya Kanta; Singh, Shiv Brat

2017-02-01

Friction Stir Welding (FSW) is a solid state joining process and is handy for welding aluminum alloys. Finite Element Method (FEM) is an important tool to predict state variables of the process but numerical simulation of FSW is highly complex due to non-linear contact interactions between tool and work piece and interdependency of displacement and temperature. In the present work, a three dimensional coupled thermo-mechanical method based on Lagrangian implicit method is proposed to study the thermal history, strain distribution and thermo-mechanical process in butt welding of Aluminum alloy 2024 using DEFORM-3D software. Workpiece is defined as rigid-visco plastic material and sticking condition between tool and work piece is defined. Adaptive re-meshing is used to tackle high mesh distortion. Effect of tool rotational and welding speed on plastic strain is studied and insight is given on asymmetric nature of FSW process. Temperature distribution on the workpiece and tool is predicted and maximum temperature is found in workpiece top surface.

Structures of FolT in substrate-bound and substrate-released conformations reveal a gating mechanism for ECF transporters

NASA Astrophysics Data System (ADS)

Zhao, Qin; Wang, Chengcheng; Wang, Chengyuan; Guo, Hui; Bao, Zhihao; Zhang, Minhua; Zhang, Peng

2015-07-01

Energy-coupling factor (ECF) transporters are a new family of ABC transporters that consist of four subunits, two cytoplasmic ATPases EcfA and EcfA' and two transmembrane proteins namely EcfS for substrate-specific binding and EcfT for energy coupling. Here, we report the 3.2-Å resolution crystal structure of the EcfS protein of a folate ECF transporter from Enterococcus faecalis-EfFolT, a close homologue of FolT from Lactobacillus brevis-LbFolT. Structural and biochemical analyses reveal the residues constituting the folate-binding pocket and determining the substrate-binding specificity. Structural comparison of the folate-bound EfFolT with the folate-free LbFolT contained in the holotransporter complex discloses significant conformational change at the L1 loop, and reveals a gating mechanism of ECF transporters in which the L1 loop of EcfS acts as a gate in the substrate binding and release.

31 CFR 202.4 - Agreement of deposit.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 31 Money and Finance: Treasury 2 2010-07-01 2010-07-01 false Agreement of deposit. 202.4 Section 202.4 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE DEPOSITARIES AND FINANCIAL AGENTS OF THE FEDERAL...

31 CFR 202.4 - Agreement of deposit.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 31 Money and Finance:Treasury 2 2012-07-01 2012-07-01 false Agreement of deposit. 202.4 Section 202.4 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY FINANCIAL MANAGEMENT SERVICE DEPOSITARIES AND FINANCIAL AGENTS OF THE FEDERAL...

Methods for Kinetic and Thermodynamic Analysis of Aminoacyl-tRNA Synthetases

PubMed Central

Francklyn, Christopher S.; First, Eric A.; Perona, John J.; Hou, Ya-Ming

2008-01-01

The accuracy of protein synthesis relies on the ability of aminoacyl-tRNA synthetases (aaRSs) to discriminate among true and near cognate substrates. To date, analysis of aaRSs function, including identification of residues of aaRS participating in amino acid and tRNA discrimination, has largely relied on the steady state kinetic pyrophosphate exchange and aminoacylation assays. Pre-steady state kinetic studies investigating a more limited set of aaRS systems have also been undertaken to assess the energetic contributions of individual enzyme-substrate interactions, particularly in the adenylation half reaction. More recently, a renewed interest in the use of rapid kinetics approaches for aaRSs has led to their application to several new aaRS systems, resulting in the identification of mechanistic differences that distinguish the two structurally distinct aaRS classes. Here, we review the techniques for thermodynamic and kinetic analysis of aaRS function. Following a brief survey of methods for the preparation of materials and for steady state kinetic analysis, this review will describe pre-steady state kinetic methods employing rapid quench and stopped-flow fluorescence for analysis of the activation and aminoacyl transfer reactions. Application of these methods to any aaRS system allows the investigator to derive detailed kinetic mechanisms for the activation and aminoacyl transfer reactions, permitting issues of substrate specificity, stereochemical mechanism, and inhibitor interaction to be addressed in a rigorous and quantitative fashion. PMID:18241792

Microstructure Evolution during Dissimilar Friction Stir Welding of AA7003-T4 and AA6060-T4

PubMed Central

Dong, Jialiang; Zhang, Datong; Zhang, Weiwen; Zhang, Wen; Qiu, Cheng

2018-01-01

In this work, the dissimilar joint of AA7003-T4 and 6060-T4 alloy has been produced by friction stir welding (FSW). The microstructure was examined by optical microscope (OM), electron back scattered diffraction (EBSD), transmission electron microscopy (TEM), and the mechanical properties of the joint were investigated. It is demonstrated that sound dissimilar joint can be produced through FSW. In the nugget; precipitations dissolve into the matrix and η′ reprecipitate subsequently; and the elongated aluminum grains are replaced by fine and equiaxed grains due to dynamic recrystallization (DRX). In the heat affected zone (HAZ), coarse β′ and η precipitates are formed and the aluminum grains are coarser as compared to the base materials. In the thermo-mechanical affected zone (TMAZ), equiaxed and elongated grains coexist due to incomplete DRX. The ultimate tensile strength of the dissimilar joint is 159.2 MPa and its elongation is 10.4%. The weak area exists in the HAZ of 6060 alloy, which is placed in the retreating side during FSW. The correlations between the microstucture and mechanical properties of the dissimilar joint are discussed. PMID:29495463

R76 in transmembrane domain 3 of the aspartate:alanine transporter AspT is involved in substrate transport.

PubMed

Suzuki, Satomi; Nanatani, Kei; Abe, Keietsu

2016-01-01

The L-aspartate:L-alanine antiporter of Tetragenococcus halophilus (AspT) possesses an arginine residue (R76) within the GxxxG motif in the central part of transmembrane domain 3 (TM3)-a residue that has been estimated to transport function. In this study, we carried out amino acid substitutions of R76 and used proteoliposome reconstitution for analyzing the transport function of each substitution. Both l-aspartate and l-alanine transport assays showed that R76K has higher activity than the AspT-WT (R76), whereas R76D and R76E have lower activity than the AspT-WT. These results suggest that R76 is involved in AspT substrate transport.

Analysis of Lightweight Materials for the AM2 System

DTIC Science & Technology

2014-06-01

and fatigue behavior in magnesium alloys . Materials Science & Engineering A (Structural Materials: Properties , Microstructure and Processing ), v 434...Table 7. Tensile properties of the alloys AA2024 or the T3 and T81 temper designations (Kuo et al . 2005...using a powder metallurgy technique, such as a standard cold compacting press and sintering process . However, the fatigue life of the liquid-based

Laser ablated high T(sub c) superconducting thin YBa2Cu3O(7-x) films on substrates suitable for microwave applications

NASA Astrophysics Data System (ADS)

Warner, J. D.; Meola, J. E.; Jenkins, K. A.; Bhasin, K. B.

1990-04-01

The development of high temperature superconducting YBa2Cu3O(7-x) thin films on substrates suitable for microwave applications is of great interest for evaluating their applications for space radar, communication, and sensor systems. Thin films of YBa2Cu3O(7-x) were formed on SrTiO3, ZrO2, MgO, and LaAlO3 substrates by laser ablation. The wavelength used was 248 nm from a KrF excimer laser. During deposition the films were heated to 600 C in a flowing oxygen environment, and required no post annealing. The low substrate temperature during deposition with no post annealing gave films which were smooth, which had their c-axis aligned to the substrates, and which had grains ranging from 0.2 to 0.5 microns in size. The films being c-axis aligned gave excellent surface resistance at 35 GHz which was lower than that of copper at 77 K. At present, LaAlO3 substrates with a dielectric constant of 22, appears suitable as a substrate for microwave and electronic applications. The films were characterized by resistance-temperature measurements, scanning electron microscopy, and x ray diffraction. The highest critical transition temperatures (T sub c) are above 89 K for films on SrTiO3 and LaAlO3, above 88 K for ZrO2, and above 86 K for MgO. The critical current density (J sub c) of the films on SrTiO3 is above 2 x 10(exp 6) amperes/sq cm at 77 K. The T(sub c) and J(sub c) are reported as a function of laser power, composition of the substrate, and temperature of the substrate during deposition.

Structural basis of Na(+)-independent and cooperative substrate/product antiport in CaiT.

PubMed

Schulze, Sabrina; Köster, Stefan; Geldmacher, Ulrike; Terwisscha van Scheltinga, Anke C; Kühlbrandt, Werner

2010-09-09

Transport of solutes across biological membranes is performed by specialized secondary transport proteins in the lipid bilayer, and is essential for life. Here we report the structures of the sodium-independent carnitine/butyrobetaine antiporter CaiT from Proteus mirabilis (PmCaiT) at 2.3-A and from Escherichia coli (EcCaiT) at 3.5-A resolution. CaiT belongs to the family of betaine/carnitine/choline transporters (BCCT), which are mostly Na(+) or H(+) dependent, whereas EcCaiT is Na(+) and H(+) independent. The three-dimensional architecture of CaiT resembles that of the Na(+)-dependent transporters LeuT and BetP, but in CaiT a methionine sulphur takes the place of the Na(+) ion to coordinate the substrate in the central transport site, accounting for Na(+)-independent transport. Both CaiT structures show the fully open, inward-facing conformation, and thus complete the set of functional states that describe the alternating access mechanism. EcCaiT contains two bound butyrobetaine substrate molecules, one in the central transport site, the other in an extracellular binding pocket. In the structure of PmCaiT, a tryptophan side chain occupies the transport site, and access to the extracellular site is blocked. Binding of both substrates to CaiT reconstituted into proteoliposomes is cooperative, with Hill coefficients up to 1.7, indicating that the extracellular site is regulatory. We propose a mechanism whereby the occupied regulatory site increases the binding affinity of the transport site and initiates substrate translocation.

Roles of tRNA in cell wall biosynthesis

PubMed Central

Dare, Kiley; Ibba, Michael

2013-01-01

Recent research into various aspects of bacterial metabolism such as cell wall and antibiotic synthesis, degradation pathways, cellular stress, and amino acid biosynthesis has elucidated roles of aminoacyl-transfer ribonucleic acid (aa-tRNA) outside of translation. Although the two enzyme families responsible for cell wall modifications, aminoacyl-phosphatidylglycerol synthases (aaPGSs) and Fem, were discovered some time ago, they have recently become of intense interest for their roles in the antimicrobial resistance of pathogenic microorganisms. The addition of positively charged amino acids to phosphatidylglycerol (PG) by aaPGSs neutralizes the lipid bilayer making the bacteria less susceptible to positively charged antimicrobial agents. Fem transferases utilize aa-tRNA to form peptide bridges that link strands of peptidoglycan. These bridges vary among the bacterial species in which they are present and play a role in resistance to antibiotics that target the cell wall. Additionally, the formation of truncated peptides results in shorter peptide bridges and loss of branched linkages which makes bacteria more susceptible to antimicrobials. A greater understanding of the structure and substrate specificity of this diverse enzymatic family is necessary to aid current efforts in designing potential bactericidal agents. These two enzyme families are linked only by the substrate with which they modify the cell wall, aa-tRNA; their structure, cell wall modification processes and the physiological changes they impart on the bacterium differ greatly. PMID:22262511

A method for predicting the fatigue life of pre-corroded 2024-T3 aluminum from breaking load tests

NASA Astrophysics Data System (ADS)

Gruenberg, Karl Martin

Characterization of material properties is necessary for design purposes and has been a topic of research for many years. Over the last several decades, much progress has been made in identifying metrics to describe fracture mechanics properties and developing procedures to measure the appropriate values. However, in the context of design, there has not been as much success in quantifying the susceptibility of a material to corrosion damage and its subsequent impact on material behavior in the framework of fracture mechanics. A natural next step in understanding the effects of corrosion damage was to develop a link between standard material test procedures and fatigue life in the presence of corrosion. Simply stated, the goal of this investigation was to formulate a cheaper and quicker method for assessing the consequences of corrosion on remaining fatigue life. For this study, breaking load specimens and fatigue specimens of a single nominal gage (0.063″) of aluminum alloy 2024-T3 were exposed to three levels of corrosion. The breaking load specimens were taken from three different material lots, and the fatigue tests were carried out at three stress levels. All failed specimens, both breaking load and fatigue specimens, were examined to characterize the damage state(s) and failure mechanism(s). Correlations between breaking load results and fatigue life results in the presence of corrosion damage were developed using a fracture mechanics foundation and the observed mechanisms of failure. Where breaking load tests showed a decrease in strength due to increased corrosion exposure, the corresponding set of fatigue tests showed a decrease in life. And where breaking load tests from different specimen orientations exhibited similar levels of strength, the corresponding set of fatigue specimens showed similar lives. The spread from shortest to longest fatigue lives among the different corrosion conditions decreased at the higher stress levels. Life predictions based

Elevated expression of CX3C chemokine receptor 1 mediates recruitment of T cells into bone marrow of patients with acquired aplastic anaemia.

PubMed

Ren, J; Hou, X Y; Ma, S H; Zhang, F K; Zhen, J H; Sun, L; Sun, Y X; Hao, Y L; Cheng, Y F; Hou, M; Xu, C G; Zhang, M H; Peng, J

2014-11-01

Acquired aplastic anaemia (AA) is a T-cell-mediated, organ-specific autoimmune disease characterized by haematopoietic stem cell destruction in the bone marrow. The exact molecular mechanism of T-cell trafficking into the bone marrow is unclear in AA. Very late activation antigen-4 (VLA-4) and CX3C chemokine receptor 1 (CX3CR1) play active roles in many autoimmune diseases. Therefore, we investigated whether VLA-4 and CX3CR1 also contribute to T-cell migration into the bone marrow in acquired AA. Expression levels of CX3CR1 and VLA-4 and their ligands [fractalkine (CX3CL1) and vascular cell adhesion molecule-1 (VCAM-1)] were examined in 63 patients with AA and 21 healthy control subjects. T-cell chemotaxis and adhesion were analysed in 17 patients with severe AA. We also prospectively evaluated the expression pattern of CX3CR1 during treatment with antithymocyte globulin plus cyclosporine in 11 patients with severe AA. The proportion of peripheral and bone marrow CD4(+) and CD8(+) T cells expressing CX3CR1 and the level of CX3CL1 was increased in patients with AA. However, there was no significant difference in VLA-4 expression or VCAM-1 levels. Functional studies demonstrated that chemotaxis towards autologous bone marrow plasma or soluble CX3CL1 was significantly higher in T cells from AA patients and could be blocked by CX3CR1 inhibitors. CX3CR1-mediated T-cell adhesion was also upregulated in these patients. The expression of CX3CR1 was associated with the efficacy of immunosuppressive therapy. The present findings demonstrate that CX3CR1 plays a pivotal role in recruitment of T cells into the bone marrow in acquired AA and is a potential therapeutic target for treatment of this disorder. © 2014 The Association for the Publication of the Journal of Internal Medicine.

Transient thermal analysis during friction stir welding between AA2014-T6 and pure copper

NASA Astrophysics Data System (ADS)

Gadhavi, A. R.; Ghetiya, N. D.; Patel, K. M.

2018-04-01

AA2xxx-Cu alloys showed larger applications in the defence sectors and in aerospace industries due to high strength to weight ratio and toughness. FSW in a butt joint configuration was carried out between AA2014-T6 and pure Copper placing AA2014 on AS and Cu on RS. Temperature profiles were observed by inserting K-type thermocouples in the mid-thickness at various locations of the plate. A sharp decrease in temperature profiles was observed on Copper side due to its higher thermal conductivity. A thermal numerical model was prepared in ANSYS to compare the simulated temperature profiles with the experimental temperature profiles and both the temperature profiles were found to be in good agreement.

Salt bridge dynamics control substrate-induced conformational change in the membrane transporter GlpT

PubMed Central

Law, Christopher J.; Almqvist, Jonas; Bernstein, Adam; Goetz, Regina M.; Huang, Yafei; Soudant, Celine; Laaksonen, Aatto; Hovmöller, Sven; Wang, Da-Neng

2008-01-01

Summary Active transport of substrates across cytoplasmic membranes is of great physiological, medical and pharmaceutical importance. The glycerol-3-phosphate (G3P) transporter (GlpT) of the E. coli inner membrane is a secondary active antiporter from the ubiquitous major facilitator superfamily that couples the import of G3P to the efflux of inorganic phosphate (Pi) down its concentration gradient. Integrating information from a novel combination of structural, molecular dynamics simulations and biochemical studies, we identify the residues involved directly in binding of substrate to the inward-facing conformation of GlpT, thus defining the structural basis for the substrate-specificity of this transporter. The substrate binding mechanism involves protonation of a histidine residue at the binding site. Furthermore, our data suggest that the formation and breaking of inter- and intradomain salt bridges control the conformational change of the transporter that accompanies substrate translocation across the membrane. The mechanism we propose may be a paradigm for organophosphate/phosphate antiporters. PMID:18395745

Damage Tolerant Design Handbook. A Compilation of Fracture and Crack- Growth Data for High-Strength Alloys. Volume 3

DTIC Science & Technology

1983-12-01

Effects on the Fatigue Crack Growth Rate of Aluminum Alloy 2024 - T351 ", University of Dayton Research Institute, Dayton, OH, Contract No. F33615-80-C-5011...ASSOCIATED WITH FIGURE 7.5.3.9 INDICATING EFFECT OF STRESS RATIO MATERIAL: ALUMINUM 2024 CONDITION: T351 ENVIRONMENT: R.T. LAB AIR DELTA K .DA/DN (10...OF STRESS INTENSITY FACTOR DATA ASSOCIATED WITH FIGURE 7.5.3.10 INDICATING EFFECT OF STRESS RATIO MATERIAL: ALUMINUM 2024

A Novel Colletotrichum graminicola Raffinose Oxidase in the AA5 Family

PubMed Central

Mollerup, Filip; Parikka, Kirsti; Koutaniemi, Sanna; Boer, Harry; Juvonen, Minna; Master, Emma; Tenkanen, Maija; Kruus, Kristiina

2017-01-01

ABSTRACT We describe here the identification and characterization of a copper radical oxidase from auxiliary activities family 5 (AA5_2) that was distinguished by showing preferential activity toward raffinose. Despite the biotechnological potential of carbohydrate oxidases from family AA5, very few members have been characterized. The gene encoding raffinose oxidase from Colletotrichum graminicola (CgRaOx; EC 1.1.3.−) was identified utilizing a bioinformatics approach based on the known modular structure of a characterized AA5_2 galactose oxidase. CgRaOx was expressed in Pichia pastoris, and the purified enzyme displayed the highest activity on the trisaccharide raffinose, whereas the activity on the disaccharide melibiose was three times lower and more than ten times lower activity was detected on d-galactose at a 300 mM substrate concentration. Thus, the substrate preference of CgRaOx was distinguished clearly from the substrate preferences of the known galactose oxidases. The site of oxidation for raffinose was studied by 1H nuclear magnetic resonance and mass spectrometry, and we confirmed that the hydroxyl group at the C-6 position was oxidized to an aldehyde and that in addition uronic acid was produced as a side product. A new electrospray ionization mass spectrometry method for the identification of C-6 oxidized products was developed, and the formation mechanism of the uronic acid was studied. CgRaOx presented a novel activity pattern in the AA5 family. IMPORTANCE Currently, there are only a few characterized members of the CAZy AA5 protein family. These enzymes are interesting from an application point of view because of their ability to utilize the cheap and abundant oxidant O2 without the requirement of complex cofactors such as FAD or NAD(P). Here, we present the identification and characterization of a novel AA5 member from Colletotrichum graminicola. As discussed in the present study, the bioinformatics approach using the modular structure of

SEM and TEM characterization of the microstructure of post-compressed TiB2/2024Al composite.

PubMed

Guo, Q; Jiang, L T; Chen, G Q; Feng, D; Sun, D L; Wu, G H

2012-02-01

In the present work, 55 vol.% TiB(2)/2024Al composites were obtained by pressure infiltration method. Compressive properties of 55 vol.% TiB(2)/2024Al composite under the strain rates of 10(-3) and 1S(-1) at different temperature were measured and microstructure of post-compressed TiB(2)/2024Al composite was characterized by scanning electron microscope (SEM) and transmission electron microscope (TEM). No trace of Al(3)Ti compound flake was found. TiB(2)-Al interface was smooth without significant reaction products, and orientation relationships ( [Formula: see text] and [Formula: see text] ) were revealed by HRTEM. Compressive strength of TiB(2)/2024Al composites decreased with temperature regardless of strain rates. The strain-rate-sensitivity of TiB(2)/2024Al composites increased with the increasing temperature. Fracture surface of specimens compressed at 25 and 250°C under 10(-3)S(-1) were characterized by furrow. Under 10(-3)S(-1), high density dislocations were formed in Al matrix when compressed at 25°C and dynamic recrystallization occurred at 250°C. Segregation of Mg and Cu on the subgrain boundary was also revealed at 550°C. Dislocations, whose density increased with temperature, were formed in TiB(2) particles under 1S(-1). Deformation of composites is affected by matrix, reinforcement and strain rate. Copyright © 2011 Elsevier Ltd. All rights reserved.

In-Situ Synthetic TiB2 Particulate Reinforced Metal Matrix Composite Coating on AA2024 Aluminum Alloy by Laser Cladding Technology

NASA Astrophysics Data System (ADS)

Xu, Jiang; Kan, Yide; Liu, Wenjin

In order to improve the wear resistance of aluminum alloy, in-situ synthesized TiB2 and Ti3B4 peritectic composite particulate reinforced metal matrix composite, formed on a 2024 aluminum alloy by laser cladding with a powder mixture of Fe-coated Boron, Ti and Al, was successfully achieved using 3-KW CW CO2 laser. The chemical composition, microstructure and phase structure of the composite clad coating were analyzed by energy dispersive X-ray spectroscopy (EDX), SEM, AFM and XRD. The typical microstructure of the composite coating is composed of TiB2, Ti3B4, Al3Ti, Al3Fe and α-Al. The surface hardness of cladding coating increases with the amount of added Fe-coated B and Ti powder which determines the amount of TiB2 and Ti3B4 peritectic composite particulate. The nanohardness and the elastic modulus at the interface of the TiB2 and Ti3B4 peritectic composite particulate/matrix were investigated using the nanoindentation technique. The results showed that the nanohardness and the reduced elastic modulus from the peritectic composite particulate to the matrix is a gradient distribution.

Microstructure and Residual Stress Distributions Under the Influence of Welding Speed in Friction Stir Welded 2024 Aluminum Alloy

NASA Astrophysics Data System (ADS)

Moghadam, Danial Ghahremani; Farhangdoost, Khalil; Nejad, Reza Masoudi

2016-06-01

Friction stir welding was conducted on 8-mm-thick plates made of AA2024-T351 aluminum alloy at tool traverse speeds between 8 and 31.5 mm/minutes and tool rotational speed between 400 and 800 rpm. Metallographic analyses and mechanical tests including hardness, tensile, residual stress, and fracture toughness tests were carried out to evaluate the microstructural and mechanical properties of the joints as a function of the process parameters. The finite element simulation of the FSW process was also performed using a thermal model. The hardness test results show that the increase in rotational speed or decrease in traverse speed of the tool would cause a decrease in weld zone hardness. The best tensile properties are obtained at rotational/traverse speed ratio between 20 and 32. Also, the longitudinal residual stress profiles were evaluated by employing X-ray diffraction method. The numerical and experimental results showed that the increase in a traverse or rotational speed would increase the residual stress of the weld zone. From the fracture toughness results, it was found that the welding process decreases the joints fracture toughness 18 to 49 pct with respect to the base metal.

Effects of plastic deformation on microstructure and superplasticity of the in situ Al3Ti/2024Al composites

NASA Astrophysics Data System (ADS)

Jiao, Lei; Yang, Yonggang; Li, Hui; Zhao, Yutao; Wang, Xiaolu

2018-05-01

In this study, the in situ Al3Ti/2024Al composites were successfully fabricated by direct melt reaction method and subjected to forging and friction stir processing (FSP) to achieve superplasticity. Then, the microstructure and superplastic tensile behavior of the composites were investigated. The results show that the reinforcement particles are broken and grains are fine after plastic processing. Particularly, the size of reinforcement particles ranges from 0.2 μm to 5 μm and the average size of fine equiaxed grains is 5 μm after FSP processing. And the superplasticity of the composites was improved apparently. The maximum elongation of 642% was obtained at 0.15 s‑1 and 510 °C for the FSP specimen, with a strain rate sensitive parameter (m) of 0.58, indicating the FSP specimen has obtained excellent high strain rate superplasticity. The strain rate sensitivity parameter, m, ranges from 0.23 to 0.58, and the activation energy was calculated to be 135.24 kJ mol‑1. All results indicated that the main superplastic deformation mechanism was grain boundary sliding (GBS) for the FSP Al3Ti/2024Al composites.

Constant and variable amplitude ultrasonic fatigue of 2024-T351 aluminium alloy at different load ratios.

PubMed

Mayer, H; Fitzka, M; Schuller, R

2013-12-01

Ultrasonic fatigue testing equipment is presented that is capable of performing constant amplitude (CA) and variable amplitude (VA) experiments at different constant load ratios. This equipment is used to study cyclic properties of aluminium alloy 2024-T351 in the high cycle fatigue (HCF) and very high cycle fatigue (VHCF) regime at load ratios R=-1 and R=0.5. CA loading does not reveal a fatigue limit below 10¹⁰ cycles. Cracks leading to VHCF failure start at broken constituent particles. Specimens that survived more than 10¹⁰ cycles at R=-1 contain non-propagating cracks of lengths below grain size. Resonance frequency and nonlinearity parameter β(rel) show changes of vibration properties of specimens at low fractions of their VHCF lifetime. VA lifetimes are measured in the HCF and VHCF regime and compared with Miner calculations. Damage sums decrease with decreasing load (and increasing mean lifetimes) and are lower for R=0.5 than R=-1. Copyright © 2013 Elsevier B.V. All rights reserved.

Polyspecific pyrrolysyl-tRNA synthetases from directed evolution.

PubMed

Guo, Li-Tao; Wang, Yane-Shih; Nakamura, Akiyoshi; Eiler, Daniel; Kavran, Jennifer M; Wong, Margaret; Kiessling, Laura L; Steitz, Thomas A; O'Donoghue, Patrick; Söll, Dieter

2014-11-25

Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNA(Pyl) have emerged as ideal translation components for genetic code innovation. Variants of the enzyme facilitate the incorporation >100 noncanonical amino acids (ncAAs) into proteins. PylRS variants were previously selected to acylate N(ε)-acetyl-Lys (AcK) onto tRNA(Pyl). Here, we examine an N(ε)-acetyl-lysyl-tRNA synthetase (AcKRS), which is polyspecific (i.e., active with a broad range of ncAAs) and 30-fold more efficient with Phe derivatives than it is with AcK. Structural and biochemical data reveal the molecular basis of polyspecificity in AcKRS and in a PylRS variant [iodo-phenylalanyl-tRNA synthetase (IFRS)] that displays both enhanced activity and substrate promiscuity over a chemical library of 313 ncAAs. IFRS, a product of directed evolution, has distinct binding modes for different ncAAs. These data indicate that in vivo selections do not produce optimally specific tRNA synthetases and suggest that translation fidelity will become an increasingly dominant factor in expanding the genetic code far beyond 20 amino acids.

Polyspecific pyrrolysyl-tRNA synthetases from directed evolution

PubMed Central

Guo, Li-Tao; Wang, Yane-Shih; Nakamura, Akiyoshi; Eiler, Daniel; Kavran, Jennifer M.; Wong, Margaret; Kiessling, Laura L.; Steitz, Thomas A.; O’Donoghue, Patrick; Söll, Dieter

2014-01-01

Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNAPyl have emerged as ideal translation components for genetic code innovation. Variants of the enzyme facilitate the incorporation >100 noncanonical amino acids (ncAAs) into proteins. PylRS variants were previously selected to acylate Nε-acetyl-Lys (AcK) onto tRNAPyl. Here, we examine an Nε-acetyl-lysyl-tRNA synthetase (AcKRS), which is polyspecific (i.e., active with a broad range of ncAAs) and 30-fold more efficient with Phe derivatives than it is with AcK. Structural and biochemical data reveal the molecular basis of polyspecificity in AcKRS and in a PylRS variant [iodo-phenylalanyl-tRNA synthetase (IFRS)] that displays both enhanced activity and substrate promiscuity over a chemical library of 313 ncAAs. IFRS, a product of directed evolution, has distinct binding modes for different ncAAs. These data indicate that in vivo selections do not produce optimally specific tRNA synthetases and suggest that translation fidelity will become an increasingly dominant factor in expanding the genetic code far beyond 20 amino acids. PMID:25385624

10 CFR 35.2024 - Records of authority and responsibilities for radiation protection programs.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 10 Energy 1 2014-01-01 2014-01-01 false Records of authority and responsibilities for radiation protection programs. 35.2024 Section 35.2024 Energy NUCLEAR REGULATORY COMMISSION MEDICAL USE OF BYPRODUCT MATERIAL Records § 35.2024 Records of authority and responsibilities for radiation protection programs. (a...

10 CFR 35.2024 - Records of authority and responsibilities for radiation protection programs.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 10 Energy 1 2013-01-01 2013-01-01 false Records of authority and responsibilities for radiation protection programs. 35.2024 Section 35.2024 Energy NUCLEAR REGULATORY COMMISSION MEDICAL USE OF BYPRODUCT MATERIAL Records § 35.2024 Records of authority and responsibilities for radiation protection programs. (a...

10 CFR 35.2024 - Records of authority and responsibilities for radiation protection programs.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 1 2010-01-01 2010-01-01 false Records of authority and responsibilities for radiation protection programs. 35.2024 Section 35.2024 Energy NUCLEAR REGULATORY COMMISSION MEDICAL USE OF BYPRODUCT MATERIAL Records § 35.2024 Records of authority and responsibilities for radiation protection programs. (a...

10 CFR 35.2024 - Records of authority and responsibilities for radiation protection programs.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 10 Energy 1 2011-01-01 2011-01-01 false Records of authority and responsibilities for radiation protection programs. 35.2024 Section 35.2024 Energy NUCLEAR REGULATORY COMMISSION MEDICAL USE OF BYPRODUCT MATERIAL Records § 35.2024 Records of authority and responsibilities for radiation protection programs. (a...

10 CFR 35.2024 - Records of authority and responsibilities for radiation protection programs.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 10 Energy 1 2012-01-01 2012-01-01 false Records of authority and responsibilities for radiation protection programs. 35.2024 Section 35.2024 Energy NUCLEAR REGULATORY COMMISSION MEDICAL USE OF BYPRODUCT MATERIAL Records § 35.2024 Records of authority and responsibilities for radiation protection programs. (a...

Effects of die quench forming on sheet thinning and 3-point bend testing of AA7075-T6

NASA Astrophysics Data System (ADS)

Kim, Samuel; Omer, Kaab; Rahmaan, Taamjeed; Butcher, Clifford; Worswick, Michael

2017-10-01

Lab-scaled AA7075 aluminum side impact beams were manufactured using the die quenching technique in which the sheet was solutionized and then quenched in-die during forming to a super saturated solid state. Sheet thinning measurements were taken at various locations throughout the length of the part and the effect of lubricant on surface scoring and material pick-up on the die was evaluated. The as-formed beams were subjected to a T6 aging treatment and then tested in three-point bending. Simulations were performed of the forming and mechanical testing experiments using the LS-DYNA finite element code. The thinning and mechanical response was predicted well.

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

Purchase of a Raman and Photoluminescence Imaging System for Characterization of Advanced Electrochemical and Electronic Materials

DTIC Science & Technology

2016-01-05

2015, Abstract #1092. The Role of Chromium (III) in the Corrosion Inhibition of AA2024-T3 By Trivalent Chromium Process Coatings by Greg Swain...to replace chromate conversion coatings and primers with more environmentally-friendly, non-chromated coatings. The Trivalent Chromium Process (TCP...coatings and primers with more environmentally-friendly, non-chromated coatings. The Trivalent Chromium Process (TCP) coating, originally developed

Bacillus thuringiensis Cry3Aa protoxin intoxication of Tenebrio molitor induces widespread changes in the expression of serine peptidase transcripts.

PubMed

Oppert, Brenda; Martynov, Alexander G; Elpidina, Elena N

2012-09-01

The yellow mealworm, Tenebrio molitor, is a pest of stored grain products and is sensitive to the Bacillus thuringiensis (Bt) Cry3Aa toxin. As digestive peptidases are a determining factor in Cry toxicity and resistance, we evaluated the expression of peptidase transcripts in the midgut of T. molitor larvae fed either a control or Cry3Aa protoxin diet for 24 h (RNA-Seq), or in larvae exposed to the protoxin for 6, 12, or 24 h (microarrays). Cysteine peptidase transcripts (9) were similar to cathepsins B, L, and K, and their expression did not vary more than 2.5-fold in control and Cry3Aa-treated larvae. Serine peptidase transcripts (48) included trypsin, chymotrypsin and chymotrypsin-like, elastase 1-like, and unclassified serine peptidases, as well as homologs lacking functional amino acids. Highly expressed trypsin and chymotrypsin transcripts were severely repressed, and most serine peptidase transcripts were expressed 2- to 15-fold lower in Cry3Aa-treated larvae. Many serine peptidase and homolog transcripts were found only in control larvae. However, expression of a few serine peptidase transcripts was increased or found only in Cry3Aa-treated larvae. Therefore, Bt intoxication significantly impacted the expression of serine peptidases, potentially important in protoxin processing, while the insect maintained the production of critical digestive cysteine peptidases. Published by Elsevier Inc.

The Disk and Jet of the Classical T Tauri Star AA Tau

NASA Technical Reports Server (NTRS)

Cox, A. W.; Grady, C. A.; Hamel, H.; Hornbeck, Jeremy; Russell, R.; Sitko, M.; Woodgate, B.

2013-01-01

Previous studies of the classical T Tauri star AA Tau have interpreted the UX Orionis-like photopolarimetric variability as being due to a warp in the inner disk caused by an inclined stellar magnetic dipolefield. We test that these effects are macroscopically observable in the inclination and alignment of the disk. We use the HST/STIS coronagraphic detection of the disk to measure the outer disk radius and inclination, and find that the inner disk is both misinclined and misaligned with respect to the outer disk. AA Tau drives a faint jet which is also misaligned with respect to the projection of the outer disk minor axis. The jet is also poorly collimated near the star. The measured inclination, 71+/-1deg, is above the inclination range suggested for stars with UX Orionis-like variability, indicating that dust grains in the disk have grown and settled toward the disk midplane.

Site-saturation engineering of lysine 47 in cyclodextrin glycosyltransferase from Paenibacillus macerans to enhance substrate specificity towards maltodextrin for enzymatic synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid (AA-2G).

PubMed

Han, Ruizhi; Liu, Long; Shin, Hyun-dong; Chen, Rachel R; Du, Guocheng; Chen, Jian

2013-07-01

In this work, the site-saturation engineering of lysine 47 in cyclodextrin glycosyltransferase (CGTase) from Paenibacillus macerans was conducted to improve the specificity of CGTase towards maltodextrin, which can be used as a cheap and easily soluble glycosyl donor for the enzymatic synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid (AA-2G) by CGTase. When using maltodextrin as glycosyl donor, four mutants K47F (lysine→ phenylalanine), K47L (lysine→ leucine), K47V (lysine→ valine) and K47W (lysine→ tryptophan) showed higher AA-2G yield as compared with that produced by the wild-type CGTase. The transformation conditions (temperature, pH and the mass ratio of L-ascorbic acid to maltodextrin) were optimized and the highest titer of AA-2G produced by the mutant K47L could reach 1.97 g/l, which was 64.2% higher than that (1.20 g/l) produced by the wild-type CGTase. The reaction kinetics analysis confirmed the enhanced maltodextrin specificity, and it was also found that compared with the wild-type CGTase, the four mutants had relatively lower cyclization activities and higher disproportionation activities, which was favorable for AA-2G synthesis. The mechanism responsible for the enhanced substrate specificity was further explored by structure modeling and it was indicated that the enhancement of maltodextrin specificity may be due to the short residue chain and the removal of hydrogen bonding interactions between the side chain of residue 47 and the sugar at -3 subsite. Here the obtained mutant CGTases, especially the K47L, has a great potential in the production of AA-2G with maltodextrin as a cheap and easily soluble substrate.

Vip3Aa induces apoptosis in cultured Spodoptera frugiperda (Sf9) cells.

PubMed

Jiang, Kun; Mei, Si-Qi; Wang, Ting-Ting; Pan, Jin-Hua; Chen, Yue-Hua; Cai, Jun

2016-09-15

The vegetative insecticidal proteins (Vip) secreted by many Bacillus thuringiensis strains during their vegetative growth stage are regarded as second generation insecticidal proteins, as they share no sequence or structural homology with known crystal insecticidal proteins (Cry) and have a broad insecticidal spectrum. Compared with insecticidal crystal proteins (ICPs), the insecticidal mechanisms of Vips have been little studied. Here we investigated the mechanism responsible for Vip3Aa toxicity in cultured insect cells. Using, flow cytometry analyzes, TUNEL staining and DNA fragmentation assays, we show that Vip3Aa can induce apoptosis in Spodoptera frugiperda (Sf9) cells and cause cells to arrest at the G2/M phase. We also show that Vip3Aa can disrupt mitochondrial membrane potential (ΔΨm), leading to the activation of Sf-caspase-1, suggesting that a mitochondrial mediated and caspase dependent pathway may be involved in Vip3Aa-induced apoptosis in Sf9 cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

High throughput assay for cytochrome P450 BM3 for screening libraries of substrates and combinatorial mutants.

PubMed

Tsotsou, Georgia Eleni; Cass, Anthony Edward George; Gilardi, Gianfranco

2002-01-01

A rapid method for identifying compounds that are potential substrates for the drug metabolising enzyme cytochrome P450 is described. The strategy is based on the detection of a degradation product of NAD(P)H oxidation during substrate turnover by the enzyme expressed in Escherichia coli cells spontaneously lysed under the experimental conditions. The performance of the method has been tested on two known substrates of the wild-type cytochrome P450 BM3, arachidonic (AA) and lauric (LA) acids, and two substrates with environmental significance, the anionic surfactant sodium dodecyl sulfate (SDS), and the solvent 1,1,2,2-tetrachloroethane (TCE). The minimal background signal given from cells expressing cytochrome P450 BM3 in the absence of added substrate is only 3% of the signal in the presence of saturating substrate. Control experiments have proven that this method is specifically detecting NADPH oxidation by catalytic turnover of P450 BM3. The assay has been adapted to a microtitre plate format and used to screen a series of furazan derivatives as potential substrates. Three derivatives were identified as substrates. The method gave a significant different signal for two isomeric furazan derivatives. All results found on the cell lysate were verified and confirmed with the purified enzyme. This strategy opens the way to automated high throughput screening of NAD(P)H-linked enzymatic activity of molecules of pharmacological and biotechnological interest and libraries of random mutants of NAD(P)H-dependent biocatalysts.

The Microjet of AA Tau

NASA Astrophysics Data System (ADS)

Cox, A. W.; Hilton, G. M.; Williger, G. M.; Grady, C. A.; Woodgate, B.

2005-12-01

The microjet of AA Tau A.W. Cox (Atholton High School, Columbia MD), G.M. Hilton (SSAI and GSFC), G.M. Williger (JHU and U. Louisville), C.A. Grady (Eureka Scientific and GSFC) B.Woodgate (NASA's GSFC) AA Tau is a classical T Tauri star with a spatially resolved disk viewed at approximately 70 degrees from pole-on. Photo-polarimetric variability of the star has been interpreted as being caused by the stellar magnetic field being inclined at 30 degrees with respect to the stellar rotation axis, producing a warp in the inner disk. Under these conditions, any jet should be less collimated than typical of T Tauri microjets, and should show signs of the jet axis precessing around the stellar rotation axis. When compared with the microjets imaged in the HST/STIS coronagraphic imaging survey, the AA Tau jet has an opening half-angle of approximately 10-15 degrees rather than the 3-5 degrees typical of the other T Tauri stars which have been coronagraphically imaged by HST/STIS. Using the HST data with ultra-narrowband imagery and long slit spectroscopy obtained with the Goddard Fabry-Perot and the Dual Imaging Spectrograph at the Apache Point Observatory 3.5m telescope, we derive the jet inclination, knot ejection epochs, and ejection frequency. We also compare the jet opening angle with model predictions. Apache Point Observatory observations with the Goddard Fabry-Perot were made through a grant of Director's Discretionary Time. Apache Point Observatory is operated by the Astrophysical Research Consortium. The GFP was supported under NASA RTOP 51-188-01-22 to GSFC. Grady is supported under NASA contract NNH05CD30C to Eureka Scientific.

Antigen-Specific CD8 Lytic Phenotype Induced by Sipuleucel-T in Hormone-Sensitive or Castration-Resistant Prostate Cancer and Association With Overall Survival.

PubMed

Antonarakis, Emmanuel S; Small, Eric J; Petrylak, Daniel; Quinn, David I; Kibel, Adam S; Chang, Nancy; Dearstyne, Erica; Harmon, Matthew; Campogan, Dwayne; Haynes, Heather; Vu, Tuyen; Sheikh, Nadeem A; Drake, Charles G

2018-06-01

Sipuleucel-T is FDA-approved for the treatment of metastatic castration-resistant prostate cancer (mCRPC) based on the IMPACT trial showing a 4.1-month benefit in median overall survival (OS) for patients receiving sipuleucel-T vs control. Although efficacy of sipuleucel-T is well-established, its mechanism remains incompletely understood. Patient samples from three sipuleucel-T trials were assessed for peripheral cellular immune responses to the immunogen PA2024 and the target antigen prostatic acid phosphatase (PAP). PAP- and PA2024-specific proliferative and cytolytic responses were characterized to delineate sipuleucel-T-induced immune responses. To quantify potential cytotoxic T lymphocyte (CTL) activity, cell-surface CD107a expression on PAP- or PA2024-specific CD8+ T cells was measured in sipuleucel-T-treated patient and healthy volunteer samples. Increased PA2024-specific CD4+ (p=0.030) and CD8+ (p=0.052) T-cell proliferation from baseline to week 6 was observed (N=14) post-sipuleucel-T, with greater magnitude of PA2024-specific responses compared to PAP. PAP- and PA2024-CTL activity (CD107a positivity) significantly increased at weeks 6 and 26 after sipuleucel-T treatment (p<0.0001; N=22). At 26 weeks post-sipuleucel-T, OS correlated with the magnitude of PAP (Pearson's R, 0.52; p=0.013) or PA2024 (Pearson's R, 0.67; p=0.0006) CTL activity. Higher PA2024-CTL activity at week 26 was significantly associated with longer OS using tertile analysis (p=0.0005; N=22), with PA2024 responses correlating with PAP responses at week 26 (R=0.90; p=1.53E -08 ). This study is the first to report PAP-specific CD8+ T-cell responses elicited by sipuleucel-T treatment. Increased and persistent potential PA2024-specific CTL activity correlated with PAP-specific CTL activity and associated with improved OS following sipuleucel-T treatment. Copyright ©2018, American Association for Cancer Research.

A Tenebrio molitor GPI-anchored alkaline phosphatase is involved in binding of Bacillus thuringiensis Cry3Aa to brush border membrane vesicles.

PubMed

Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Bravo, Alejandra; Soberón, Mario

2013-03-01

Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders. Copyright © 2012 Elsevier Inc. All rights reserved.

Influence of FSW pin tool geometry on plastic flow of AA7075 T651

NASA Astrophysics Data System (ADS)

Lertora, Enrico; Mandolfino, Chiara; Gambaro, Carla

2016-10-01

In this paper the behaviour of the plastic flow during Friction Stir Welding of AA7075 T651 plates, realized with different shaped tools, has been investigated. In particular, the influence of the shape of three tools was studied using copper strips placed along the welds. After welding, radiography and metallurgical analysis were used in order to investigate the marker movement and its fragmentation.

Insights into the Structure of the Vip3Aa Insecticidal Protein by Protease Digestion Analysis

PubMed Central

Bel, Yolanda; Banyuls, Núria; Chakroun, Maissa; Escriche, Baltasar; Ferré, Juan

2017-01-01

Vip3 proteins are secretable proteins from Bacillus thuringiensis whose mode of action is still poorly understood. In this study, the activation process for Vip3 proteins was closely examined in order to better understand the Vip3Aa protein stability and to shed light on its structure. The Vip3Aa protoxin (of 89 kDa) was treated with trypsin at concentrations from 1:100 to 120:100 (trypsin:Vip3A, w:w). If the action of trypsin was not properly neutralized, the results of SDS-PAGE analysis (as well as those with Agrotis ipsilon midgut juice) equivocally indicated that the protoxin could be completely processed. However, when the proteolytic reaction was efficiently stopped, it was revealed that the protoxin was only cleaved at a primary cleavage site, regardless of the amount of trypsin used. The 66 kDa and the 19 kDa peptides generated by the proteases co-eluted after gel filtration chromatography, indicating that they remain together after cleavage. The 66 kDa fragment was found to be extremely resistant to proteases. The trypsin treatment of the protoxin in the presence of SDS revealed the presence of secondary cleavage sites at S-509, and presumably at T-466 and V-372, rendering C-terminal fragments of approximately 29, 32, and 42 kDa, respectively. The fact that the predicted secondary structure of the Vip3Aa protein shows a cluster of beta sheets in the C-terminal region of the protein might be the reason behind the higher stability to proteases compared to the rest of the protein, which is mainly composed of alpha helices. PMID:28387713

Through-thickness recrystallization characteristics of a laminated AA3xxx–AA6xxx aluminum alloy system

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liao, L.H., E-mail: l2liao@uwaterloo.ca; Jin, H.; Gallerneault, M.

2015-03-15

The through-thickness annealing behavior of a laminated AA3xxx–AA6xxx alloy system at 300 °C has been studied by scanning electron microscopy, electron backscatter diffraction analysis, electron probe micro-analysis, differential scanning calorimetry, and hardness measurement. Results show that the recrystallization process starts at the interface region between the AA3xxx (clad) and AA6xxx (core) layers. Subsequently, the recrystallization process front progresses into the core layer, while the clad layer is the last region to recrystallize. It is also found that precipitation precedes recrystallization in the entire laminate at the investigated temperature. The preferential onset of recrystallization at the interface region is attributed tomore » the net driving pressure being the highest in this region. The factors that lead to such enhanced net driving pressure are (a) deformation incompatibility between the two alloy layers, (b) lower solute content of the interface, which also leads to lower volume fraction of precipitates, and (c) an accelerated rate of precipitate coarsening due to the presence of a higher density of dislocations. The gradual progress of recrystallization from the interface towards the core layer is dictated by precipitate coarsening and the dependence of its rate on the density of deformation-induced dislocations. The lower driving pressure due to lower work hardening capacity, high solute drag pressure due to Mn, and additional Zener drag from precipitates that form due to solute redistribution during annealing explain the late initiation of recrystallization in the clad layer. - Highlights: • The through-thickness recrystallization of a laminated system is investigated. • The early onset of recrystallization at the interface is discussed. • The effects of precipitation and coarsening on recrystallization are analyzed.« less

Selection and characterization of resistance to the Vip3Aa20 protein from Bacillus thuringiensis in Spodoptera frugiperda.

PubMed

Bernardi, Oderlei; Bernardi, Daniel; Horikoshi, Renato J; Okuma, Daniela M; Miraldo, Leonardo L; Fatoretto, Julio; Medeiros, Fernanda Cl; Burd, Tony; Omoto, Celso

2016-09-01

Spodoptera frugiperda is one the main target pests of maize events expressing Vip3Aa20 protein from Bacillus thuringiensis (Bt) in Brazil. In this study, we selected a resistant strain of S. frugiperda on Bt maize expressing Vip3Aa20 protein and characterized the inheritance and fitness costs of the resistance. The resistance ratio of the Vip3Aa20-resistant strain of S. frugiperda was >3200-fold. Neonates of the Vip3Aa20-resistant strain were able to survive and emerge as fertile adults on Vip3Aa20 maize, while larvae from susceptible and heterozygous strains did not survive. The inheritance of Vip3Aa20 resistance was autosomal recessive and monogenic. Life history studies to investigate fitness cost revealed an 11% reduction in the survival rate until adult stage and a ∼50% lower reproductive rate of the Vip3Aa20-resistant strain compared with susceptible and heterozygous strains. This is the first characterization of S. frugiperda resistance to Vip3Aa protein. Our results provide useful information for resistance management programs designed to prevent or delay resistance evolution to Vip3Aa proteins in S. frugiperda. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Effect of Localized Corrosion on Fatigue-Crack Growth in 2524-T3 and 2198-T851 Aluminum Alloys Used as Aircraft Materials

NASA Astrophysics Data System (ADS)

Moreto, J. A.; Broday, E. E.; Rossino, L. S.; Fernandes, J. C. S.; Bose Filho, W. W.

2018-03-01

Corrosion and fatigue of aluminum alloys are major issues for the in-service life assessment of aircraft structures and for the management of aging air fleets. The aim of this work was to evaluate the effect of localized corrosion on fatigue crack growth (FCG) resistance of the AA2198-T851 Al-Li alloy (Solution Heat Treated, Cold Worked, and Artificially Aged), comparing it with the FCG resistance of AA2524-T3 (Solution Heat Treated and Cold Worked), considering the effect of seawater fog environment. Before fatigue tests, the corrosion behavior of 2198-T851 and 2524-T3 aluminum alloys was verified using open circuit potential and potentiodynamic polarization techniques. Fatigue in air and corrosion fatigue tests were performed applying a stress ratio (R) of 0.1, 15 Hz (air) and 0.1 Hz (seawater fog) frequencies, using a sinusoidal waveform in all cases. The results showed that the localized characteristics of the 2198-T851 and 2524-T3 aluminum alloys are essentially related to the existence of intermetallic compounds, which, due to their different nature, may be cathodic or anodic in relation to the aluminum matrix. The corrosive medium has affected the FCG rate of both aluminum alloys, in a quite similar way.

An alternate to chromate conversion coatings for the corrosion protection of aluminum 2024-T3

NASA Astrophysics Data System (ADS)

Guo, Ruiguang

Corrosion of high-strength aluminum alloys used for airspace application is an expensive and serious problem. The most significant environmental factor contributing to the corrosion of these alloys is water condensed from humid air and contaminated with soluble chloride salts. The Al 2024 series used for aircraft are particularly susceptible to corrosion in aqueous chloride solutions due to alloying constituents such as copper and other impurities. Chromates are efficient inhibitors of corrosion of aluminum in near neutral aqueous environments containing aggressive anions such as chlorides. Usually, aluminum alloys are initially protected by chromate conversion coatings. Additional polymer coatings are sometimes added during exposure to corrosive atmospheres such as marine environments. Although chromate coatings are widely used, they require the use of noxious solutions, so they have always presented effluent disposal problems. There are health and safety concerns over the use of chromates due to their toxicity and carcinogenic nature and, as a consequence, the environmental and health risks associated with the use of such coatings will be restricted in the future. It was these health and safety concerns that led to the development of alternative non-toxic coating processes with comparable adhesion properties and corrosion protection. A variety of process technologies are under development and are vying for acceptance in industrial markets. As an alternate conversion coating, a new titanate conversion coating was systematically researched and developed. Research concentrated on producing passive surfaces from a simple titanate solution using an immersion process. The corrosion resistance of the treated surface has been evaluated using simple, rapid electrochemical techniques as well as a more long-term salt spray test. Passivation by titanate conversion treatment exhibits many similarities to chromate conversion treatment. Based on this study of corrosion

AAS 228: Day 3 afternoon

NASA Astrophysics Data System (ADS)

Kohler, Susanna

2016-06-01

?).Peebles: expect surprises and think outside the box yes! #aas228 Kevin Schawinski (@kevinschawinski) June 15, 2016His comparison of local vs. high-redshift cosmology in terms of scales of problems was outstanding, and he encouraged the audience and the community to keep an open mind. After all, in Sean Carrolls words, we do live in a preposterous universe, and the time is almost right to explain it all.This The Limits of Scientific Cosmology series at #aas228 has been absolutely fantastic. @AAS_Office please put videos on youtube! Kevin Schawinski (@kevinschawinski) June 15, 2016Press Conference Black Holes and Gamma-Ray Bursts (by Susanna Kohler)The final press conference of the meeting covered three topics from the categories of black holes and gamma-ray bursts.Fred Rasio (CIERA/Northwestern University) opened the conference with a discussion of how the systems that LIGO detected might have formed. There are two primary models for how these black-hole binaries are created. In the first, they start out as binary systems of two massive stars. If the binary survives the process of both stars collapsing into black holes, then a binary black hole system results.Rasio focused on the second theory, in which that black holes are formed in dense stellar clusters. These black holes then sink (via dynamical friction) to the centers of the clusters like dust particles settling on the floor of a room, where they form binaries in a black hole mosh pit eventually getting kicked out of the cluster by dynamical interactions. You can read more about their research in the press release here.Rasio presents a pretty awesome animation of this black hole mosh pit. https://t.co/AkewctI7uK#aas228 astrobites (@astrobites) June 15, 2016The effects of a spinning black hole on the spacetime around it. [J.P. Eekels J.M. Overduin]Next up was Richard Henry (Johns Hopkins University), who spoke about the internal structure of spinning black holes (known as Kerr black holes). Because no light can escape from

Differential Regulation of ERK1/2 and mTORC1 Through T1R1/T1R3 in MIN6 Cells.

PubMed

Wauson, Eric M; Guerra, Marcy L; Dyachok, Julia; McGlynn, Kathleen; Giles, Jennifer; Ross, Elliott M; Cobb, Melanie H

2015-08-01

The MAPKs ERK1/2 respond to nutrients and other insulin secretagogues in pancreatic β-cells and mediate nutrient-dependent insulin gene transcription. Nutrients also stimulate the mechanistic target of rapamycin complex 1 (mTORC1) to regulate protein synthesis. We showed previously that activation of both ERK1/2 and mTORC1 in the MIN6 pancreatic β-cell-derived line by extracellular amino acids (AAs) is at least in part mediated by the heterodimeric T1R1/T1R3, a G protein-coupled receptor. We show here that AAs differentially activate these two signaling pathways in MIN6 cells. Pretreatment with pertussis toxin did not prevent the activation of either ERK1/2 or mTORC1 by AAs, indicating that G(I) is not central to either pathway. Although glucagon-like peptide 1, an agonist for a G(s-)coupled receptor, activated ERK1/2 well and mTORC1 to a small extent, AAs had no effect on cytosolic cAMP accumulation. Ca(2+) entry is required for ERK1/2 activation by AAs but is dispensable for AA activation of mTORC1. Pretreatment with UBO-QIC, a selective G(q) inhibitor, reduced the activation of ERK1/2 but had little effect on the activation of mTORC1 by AAs, suggesting a differential requirement for G(q). Inhibition of G(12/13) by the overexpression of the regulator of G protein signaling domain of p115 ρ-guanine nucleotide exchange factor had no effect on mTORC1 activation by AAs, suggesting that these G proteins are also not involved. We conclude that AAs regulate ERK1/2 and mTORC1 through distinct signaling pathways.

Differential Regulation of ERK1/2 and mTORC1 Through T1R1/T1R3 in MIN6 Cells

PubMed Central

Wauson, Eric M.; Guerra, Marcy L.; Dyachok, Julia; McGlynn, Kathleen; Giles, Jennifer; Ross, Elliott M.

2015-01-01

The MAPKs ERK1/2 respond to nutrients and other insulin secretagogues in pancreatic β-cells and mediate nutrient-dependent insulin gene transcription. Nutrients also stimulate the mechanistic target of rapamycin complex 1 (mTORC1) to regulate protein synthesis. We showed previously that activation of both ERK1/2 and mTORC1 in the MIN6 pancreatic β-cell-derived line by extracellular amino acids (AAs) is at least in part mediated by the heterodimeric T1R1/T1R3, a G protein-coupled receptor. We show here that AAs differentially activate these two signaling pathways in MIN6 cells. Pretreatment with pertussis toxin did not prevent the activation of either ERK1/2 or mTORC1 by AAs, indicating that Gi is not central to either pathway. Although glucagon-like peptide 1, an agonist for a Gs-coupled receptor, activated ERK1/2 well and mTORC1 to a small extent, AAs had no effect on cytosolic cAMP accumulation. Ca2+ entry is required for ERK1/2 activation by AAs but is dispensable for AA activation of mTORC1. Pretreatment with UBO-QIC, a selective Gq inhibitor, reduced the activation of ERK1/2 but had little effect on the activation of mTORC1 by AAs, suggesting a differential requirement for Gq. Inhibition of G12/13 by the overexpression of the regulator of G protein signaling domain of p115 ρ-guanine nucleotide exchange factor had no effect on mTORC1 activation by AAs, suggesting that these G proteins are also not involved. We conclude that AAs regulate ERK1/2 and mTORC1 through distinct signaling pathways. PMID:26168033

Substrate-modulated unwinding of transmembrane helices in the NSS transporter LeuT.

PubMed

Merkle, Patrick S; Gotfryd, Kamil; Cuendet, Michel A; Leth-Espensen, Katrine Z; Gether, Ulrik; Loland, Claus J; Rand, Kasper D

2018-05-01

LeuT, a prokaryotic member of the neurotransmitter:sodium symporter (NSS) family, is an established structural model for mammalian NSS counterparts. We investigate the substrate translocation mechanism of LeuT by measuring the solution-phase structural dynamics of the transporter in distinct functional states by hydrogen/deuterium exchange mass spectrometry (HDX-MS). Our HDX-MS data pinpoint LeuT segments involved in substrate transport and reveal for the first time a comprehensive and detailed view of the dynamics associated with transition of the transporter between outward- and inward-facing configurations in a Na + - and K + -dependent manner. The results suggest that partial unwinding of transmembrane helices 1/5/6/7 drives LeuT from a substrate-bound, outward-facing occluded conformation toward an inward-facing open state. These hitherto unknown, large-scale conformational changes in functionally important transmembrane segments, observed for LeuT in detergent-solubilized form and when embedded in a native-like phospholipid bilayer, could be of physiological relevance for the translocation process.

Joining sheet aluminum AA6061-T4 to cast magnesium AM60B by vaporizing foil actuator welding: Input energy, interface, and strength

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Bert; Vivek, Anupam; Daehn, Glenn S.

Dissimilar joining of sheet aluminum AA6061-T4 to cast magnesium AM60B was achieved by vaporizing foil actuator welding (VFAW). Three input energy levels were used (6, 8, and 10 kJ), and as a trend, higher input energies resulted in progressively higher flyer velocities, more pronounced interfacial wavy features, larger weld zones, higher peel strengths, and higher peel energies. In all cases, weld cross section revealed a soundly bonded interface characterized by well-developed wavy features and lack of voids and continuous layers of intermetallic compounds (IMCs). At 10 kJ input energy, flyer speed of 820 m/s, peel strength of 22.4 N/mm, andmore » peel energy of 5.2 J were obtained. In lap-shear, failure occurred in AA6061- T4 flyer at 97% of the base material’s peak tensile load. Peel samples failed along the weld interface, and the AM60B-side of the fracture surface showed thin, evenly-spaced lines of Al residuals which had been torn out of the base AA6061-T4 in a ductile fashion and transferred over to the AM60B side, indicating very strong AA6061-T4/AM60B bond in these areas. Furthermore, this work demonstrates VFAW’s capability in joining dissimilar lightweight metals such as Al/Mg, which is expected to be a great enabler in the ongoing push for vehicle weight reduction.« less

Joining sheet aluminum AA6061-T4 to cast magnesium AM60B by vaporizing foil actuator welding: Input energy, interface, and strength

DOE PAGES

Liu, Bert; Vivek, Anupam; Daehn, Glenn S.

2017-09-19

Dissimilar joining of sheet aluminum AA6061-T4 to cast magnesium AM60B was achieved by vaporizing foil actuator welding (VFAW). Three input energy levels were used (6, 8, and 10 kJ), and as a trend, higher input energies resulted in progressively higher flyer velocities, more pronounced interfacial wavy features, larger weld zones, higher peel strengths, and higher peel energies. In all cases, weld cross section revealed a soundly bonded interface characterized by well-developed wavy features and lack of voids and continuous layers of intermetallic compounds (IMCs). At 10 kJ input energy, flyer speed of 820 m/s, peel strength of 22.4 N/mm, andmore » peel energy of 5.2 J were obtained. In lap-shear, failure occurred in AA6061- T4 flyer at 97% of the base material’s peak tensile load. Peel samples failed along the weld interface, and the AM60B-side of the fracture surface showed thin, evenly-spaced lines of Al residuals which had been torn out of the base AA6061-T4 in a ductile fashion and transferred over to the AM60B side, indicating very strong AA6061-T4/AM60B bond in these areas. Furthermore, this work demonstrates VFAW’s capability in joining dissimilar lightweight metals such as Al/Mg, which is expected to be a great enabler in the ongoing push for vehicle weight reduction.« less

Computational Insights into the High-Fidelity Catalysis of Aminoacyl-tRNA Synthetases

NASA Astrophysics Data System (ADS)

Aboelnga, Mohamed M.

Obtaining insights into the catalytic function of enzymes is an important area of research due to their widespread applications in the biotechnology and pharmaceutical industries. Among these enzymes, the aminoacyl-tRNA synthetases (aaRSs) are known for their remarkable fidelity in catalyzing the aminoacylation reactions of tRNA in protein biosynthesis. Despite the exceptional execution of this critical function, mechanistic details of the reactions catalyzed by aminoacyl-tRNA synthetases remain elusive demonstrating the obvious need to explore their remarkable chemistry. During the PhD studies reported in this thesis the mechanism of aminoacylation, pre?transfer editing and post?transfer editing catalyzed by different aaRS have been established using multi-scale computational enzymology. In the first two chapters a detailed information about aaRS and the addressed questions was given in addition to an overview of the used computational methodology currently used to investigate the enzymatic mechanisms. The aminoacylation mechanism of threonine by Threonyl-tRNA synthetases, glutamine by Glutaminyl-tRNA synthetases and glutamate by Glutamyl-tRNA synthetases have been clearly unveiled in chapter 3 and 4. Also, valuable information regarding the role of cofactors and active site residues has been obtained. While investigating the post-transfer editing mechanisms, which proceed in a remote and distinct active site, two different scenarios were experimentally suggested for two types of threonyl-tRNA synthetase species to correct the misacylation of the structurally related serine. We explored these two mechanisms as in chapters 5 and 6. Moreover, the synthetic site in which the aminoacylation reaction is catalyzed, is also responsible for a second type of proofreading reaction called pre-transfer editing mechanism. In chapter 7, this latter mechanism has been elucidated for both Seryl-tRNA synthetases and Isoleucyl-tRNA synthetases against their non-cognate substrates

Poly(2,5-bis(N-Methyl-N-Hexylamino)Phenylene Vinylene) (BAM-PPV) as Pretreatment Coating for Aerospace Applications: Laboratory and Field Studies

PubMed Central

Zarras, Peter; Buhrmaster, Diane; Webber, Cindy; Anderson, Nicole; Stenger-Smith, John D.; Goodman, Paul A.

2014-01-01

In this study, an electroactive polymer (EAP), poly(2,5-bis(N-methyl-N-hexylamino)phenylene vinylene) (BAM-PPV) was investigated as a potential alternative surface pretreatment for hexavalent chromium (Cr(VI))-based aerospace coatings. BAM-PPV was tested as a pretreatment coating on an aerospace aluminum alloy (AA2024-T3) substrate in combination with a non-Cr(VI) epoxy primer and a polyurethane Advanced Performance Coating (APC) topcoat. This testing was undertaken to determine BAM-PPV’s adhesion, corrosion-inhibition, compatibility and survivability in laboratory testing and during outdoor field-testing. BAM-PPV showed excellent adhesion and acceptable corrosion performance in laboratory testing. The BAM-PPV aerospace coating system (BAM-PPV, non-Cr(VI) epoxy primer and polyurethane APC topcoat) was field tested for one year on the rear hatch door of the United States Air Force C-5 cargo plane. After one year of field testing there was no evidence of delamination or corrosion of the BAM-PPV aerospace coating system. PMID:28788292

Poly(2,5-bis(N-Methyl-N-Hexylamino)Phenylene Vinylene) (BAM-PPV) as Pretreatment Coating for Aerospace Applications: Laboratory and Field Studies.

PubMed

Zarras, Peter; Buhrmaster, Diane; Webber, Cindy; Anderson, Nicole; Stenger-Smith, John D; Goodman, Paul A

2014-12-17

In this study, an electroactive polymer (EAP), poly(2,5-bis( N -methyl- N -hexylamino)phenylene vinylene) (BAM-PPV) was investigated as a potential alternative surface pretreatment for hexavalent chromium (Cr(VI))-based aerospace coatings. BAM-PPV was tested as a pretreatment coating on an aerospace aluminum alloy (AA2024-T3) substrate in combination with a non-Cr(VI) epoxy primer and a polyurethane Advanced Performance Coating (APC) topcoat. This testing was undertaken to determine BAM-PPV's adhesion, corrosion-inhibition, compatibility and survivability in laboratory testing and during outdoor field-testing. BAM-PPV showed excellent adhesion and acceptable corrosion performance in laboratory testing. The BAM-PPV aerospace coating system (BAM-PPV, non-Cr(VI) epoxy primer and polyurethane APC topcoat) was field tested for one year on the rear hatch door of the United States Air Force C-5 cargo plane. After one year of field testing there was no evidence of delamination or corrosion of the BAM-PPV aerospace coating system.

Electrochemical investigation of powder coatings and their application to magnesium-rich primers for corrosion protection

NASA Astrophysics Data System (ADS)

Orgon, Casey Roy

Corrosion is the decomposition of metal and metal alloys which threatens the integrity of man-made structures. One of the more efficient methods of delaying the corrosion process in metals is by coatings. In this work, the durability of two polyester powder coatings were investigated for corrosion protection of AA-2024-T3. Polyester powder coatings crosslinked by either triglycidyl isocyanurate (TGIC) or beta-hydroxyalkyl amide (HAA) compounds were prepared and investigated for barrier protection of metal substrates by electrochemical impedance spectroscopy (EIS). Polyester-TGIC coatings were found to provide better long-term protection, which can be attributed to the increased mechanical strength and higher concentration of crosslinking in the coating films. Additionally, the polyester powder coatings, along with a fusion bonded epoxy (FBE) were investigated for their compatibility as a topcoat for magnesium-rich primers (MgRP). Under proper application conditions, powder topcoats were successfully applied to cured MgRP while corrosion protection mechanisms of each system were maintained.

AAS 227: Day 3

NASA Astrophysics Data System (ADS)

Kohler, Susanna

2016-01-01

Editors Note:This week were at the 227th AAS Meeting in Kissimmee, FL. Along with several fellow authors from astrobites.com, I will bewritingupdates on selectedevents at themeeting and posting at the end of each day. Follow along here or atastrobites.com, or catch ourlive-tweeted updates from the@astrobites Twitter account. The usual posting schedule for AAS Nova will resumenext week.Welcome to Day 3 of the winter American Astronomical Society (AAS) meeting in Kissimmee! Several of us are attending the conference this year, and we will report highlights from each day here on astrobites. If youd like to see more timely updates during the day, we encourage you to follow @astrobites on twitter or search the #aas227 hashtag.Henry Norris Russell Lecture: Viewing the Universe with Infrared Eyes: The Spitzer Space Telescope (by Erika Nesvold)The Henry Norris Russell Award is the highest honor given by the AAS, for a lifetime of eminence in astronomy research. This years award went to Giovanni Fazio of the Harvard-Smithsonian Center for Astrophysics. Fazio became a leader in gamma ray astronomy before switching mid-career to the study of infrared astronomy, and he gave his award lecture on the latter subject, specifically on the Spitzer Space Telescope, one of the most successful infrared telescopes of all time.Artists rendering of the Spitzer space telescope. [NASA/JPL-Caltech]Spitzer has been operating for more than twelve years, and has resulted in over six thousand papers in refereed journals in that time. The telescope sits in an Earth-trailing orbit around the Sun, and is now farther from the Earth (1.4 AU) than the Earth is from the Sun. Fazio gave the audience a fascinating overview of the science done by Spitzer over more than a decade. One of the most productive areas of research for Spitzer is the study of exoplanets, which hadnt even been discovered when the Spitzer Telescope was first conceived. Spitzers high sensitivity and ability to observe exoplanets over

Influence of surface pretreatments on the quality of trivalent chromium process coatings on aluminum alloy

NASA Astrophysics Data System (ADS)

Viroulaud, Rémi; Światowska, Jolanta; Seyeux, Antoine; Zanna, Sandrine; Tardelli, Joffrey; Marcus, Philippe

2017-11-01

The effects of surface pretreatments (degreasing and pickling) on the characteristics of the Trivalent Chromium Process (TCP) coating on pure aluminum and on AA2024-T351 aluminum alloy were investigated here by means of surface sensitive techniques: X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). The XPS and ToF-SIMS results evidence that the TCP coating homogeneity is strongly dependent on the pretreatment process used. The TCP coverage factor, calculated from XPS results, is significantly lower, on both pure aluminum and AA2024-T351 alloy surface, when a pickling step is applied. One of the main effects of pickling pretreatment is strong metallic copper enrichment at the surface of the 2024 alloy, associated with chemical dissolution of Al-Cu intermetallic particles. However, it is evidenced here, that the copper enrichment is not detrimental for the quality of the TCP coating. The coating failure, observed when the pickling step is applied, can be assigned to a faster kinetics of the coating growth leading to formation of thicker conversion coating more susceptible to cracking or to the localized presence of aluminum fluoride species leading to the appearance of coating defects or detachment.

Synthesis and characterization of nanocrystalline Al 2024-B4C composite powders by mechanical alloying

NASA Astrophysics Data System (ADS)

Varol, T.; Canakci, A.

2013-06-01

In the present work, the effect of milling parameters on the morphology and microstructure of nanostructure Al2024-B4C composite powders obtained by mechanical alloying (MA) was studied. The effects of milling time and B4C content on the morphology, microstructure and particle size of nanostructure Al2024-B4C composite powders have been investigated. Different amounts of B4C particles (0, 5, 10 and 20 wt.%) were mixed with Al2024 powders and milled in a planetary ball mill for 30, 60, 120, 300, 420 and 600 min. Al 2024-B4C composite powders were characterized using a scanning electron microscope (SEM), laser particle-size analyzer, X-ray diffraction analysis (XRD) and the Vickers microhardness test. The results showed that the nanostructure Al2024-B4C composite powders were produced when they were milled for 600 min. The size of composite powder in the milled powder mixture was affected by the milling time and content of B4C particles. Moreover, it was observed that when MA reached a steady state, the properties of composite powders were stabilized.

Imaging the Disk and Jet of the Classical T Tauri Star AA Tau

NASA Astrophysics Data System (ADS)

Cox, Andrew; Grady, C.; Hammel, H. B.; Hornbeck, J.; Russell, R. W.; Sitko, M. L.; Woodgate, B. E.

2013-01-01

Previous studies of the classical T Tauri star AA Tau have interpreted the UX Orionis-like photo-polarimetric variability as being due to a warp in the inner disk caused by an inclined stellar magnetic dipole field. We test that these effects are macroscopically observable in the inclination and alignment of the disk. We use HST/STIS coronagraphic imagery to measure the V magnitude of the star for both STIS corona graphic observations, compare these data with optical photometry in the literature and find that unlike other classical T Tauri stars observed on the same HST program, the disk is most robustly detected at optical minimum light. We measure the outer disk radius, major axis position angle, and disk inclination, and find that the inner disk, as reported in the literature, is both mis-inclined and misaligned with respect to the outer disk. AA Tau drives a faint jet which is also misaligned with respect to the projection of the outer disk minor axis and which is poorly collimated near the star. The measured outer disk inclination, 71±1 degrees, is out of the inclination band suggested for stars with UX Orionis-like variability where no grain growth has occurred in the disk. The faintness of the disk, the small disk size, and visibility of the star and despite the high inclination, all indicate that the disk must have experienced grain growth and settling toward the disk midplane, which we verify by comparing the observed disk with model imagery from the literature.

40 CFR 35.2024 - Combined sewer overflows.

Code of Federal Regulations, 2014 CFR

2014-07-01

... ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works § 35.2024 Combined sewer... project priority list, it addresses impaired uses in priority water quality areas which are due to the... must demonstrate to the Administrator that the water quality goals of the Act will not be achieved...

40 CFR 35.2024 - Combined sewer overflows.

Code of Federal Regulations, 2012 CFR

2012-07-01

... ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works § 35.2024 Combined sewer... project priority list, it addresses impaired uses in priority water quality areas which are due to the... must demonstrate to the Administrator that the water quality goals of the Act will not be achieved...

40 CFR 35.2024 - Combined sewer overflows.

Code of Federal Regulations, 2011 CFR

2011-07-01

... ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works § 35.2024 Combined sewer... project priority list, it addresses impaired uses in priority water quality areas which are due to the... must demonstrate to the Administrator that the water quality goals of the Act will not be achieved...

Substrate sequence selectivity of APOBEC3A implicates intra-DNA interactions.

PubMed

Silvas, Tania V; Hou, Shurong; Myint, Wazo; Nalivaika, Ellen; Somasundaran, Mohan; Kelch, Brian A; Matsuo, Hiroshi; Kurt Yilmaz, Nese; Schiffer, Celia A

2018-05-14

The APOBEC3 (A3) family of human cytidine deaminases is renowned for providing a first line of defense against many exogenous and endogenous retroviruses. However, the ability of these proteins to deaminate deoxycytidines in ssDNA makes A3s a double-edged sword. When overexpressed, A3s can mutate endogenous genomic DNA resulting in a variety of cancers. Although the sequence context for mutating DNA varies among A3s, the mechanism for substrate sequence specificity is not well understood. To characterize substrate specificity of A3A, a systematic approach was used to quantify the affinity for substrate as a function of sequence context, length, secondary structure, and solution pH. We identified the A3A ssDNA binding motif as (T/C)TC(A/G), which correlated with enzymatic activity. We also validated that A3A binds RNA in a sequence specific manner. A3A bound tighter to substrate binding motif within a hairpin loop compared to linear oligonucleotide, suggesting A3A affinity is modulated by substrate structure. Based on these findings and previously published A3A-ssDNA co-crystal structures, we propose a new model with intra-DNA interactions for the molecular mechanism underlying A3A sequence preference. Overall, the sequence and structural preferences identified for A3A leads to a new paradigm for identifying A3A's involvement in mutation of endogenous or exogenous DNA.

Evaluation of Fatigue Behavior and Surface Characteristics of Aluminum Alloy 2024 T6 After Electric Discharge Machining

NASA Astrophysics Data System (ADS)

Mehmood, Shahid; Shah, Masood; Pasha, Riffat Asim; Sultan, Amir

2017-10-01

The effect of electric discharge machining (EDM) on surface quality and consequently on the fatigue performance of Al 2024 T6 is investigated. Five levels of discharge current are analyzed, while all other electrical and nonelectrical parameters are kept constant. At each discharge current level, dog-bone specimens are machined by generating a peripheral notch at the center. The fatigue tests are performed on four-point rotating bending machine at room temperature. For comparison purposes, fatigue tests are also performed on the conventionally machined specimens. Linearized SN curves for 95% failure probability and with four different confidence levels (75, 90, 95 and 99%) are plotted for each discharge current level as well as for conventionally machined specimens. These plots show that the electric discharge machined (EDMed) specimens give inferior fatigue behavior as compared to conventionally machined specimen. Moreover, discharge current inversely affects the fatigue life, and this influence is highly pronounced at lower stresses. The EDMed surfaces are characterized by surface properties that could be responsible for change in fatigue life such as surface morphology, surface roughness, white layer thickness, microhardness and residual stresses. It is found that all these surface properties are affected by changing discharge current level. However, change in fatigue life by discharge current could not be associated independently to any single surface property.

A study comparing changes in loading conditions of an extended service life system using aluminum 2024-T351

NASA Astrophysics Data System (ADS)

Beal, Roger Zack

The current fiscally austere environment prevalent in the military and industry is driving extreme measures to save money. In the United States Air Force, this has driven enormous efforts to trim sustainment spending on extended life aircraft. The challenge to the aerospace engineer is to ensure flight safety in the midst of this economic pressure. One method of cutting costs is to increase the time an aircraft is in service by delaying the point when the aircraft is taken out of service for depot maintenance. To ensure flight safety, in depth fatigue and fracture analysis needs to be accomplished to assess increasing the inspection interval. The purpose of this study was to determine the sensitivity of Aluminum 2024-T351 alloy, a common material used in tension dominated aerospace applications, to two different loading spectra---one that is aggressive and the other that is benign. This was accomplished by conducting five different combinations of the two spectra, developing computer simulations using the AFGROW software and comparing with the measured data. The results showed that the material demonstrated significantly different behavior between the two spectra. These results provide a valuable tool for the aerospace engineer for fatigue life prediction and inspection interval evaluation.

p62-mediated autophagy affects nutrition-dependent insulin receptor substrate-1 dynamics in 3T3-L1 preadipocytes.

PubMed

Igawa, Hirobumi; Kikuchi, Akihiro; Misu, Hirofumi; Ishii, Kiyo-Aki; Kaneko, Shuichi; Takamura, Toshinari

2018-05-22

Previous studies have shown that the organism's nutritional status changes the protein levels of insulin receptor substrate 1 (IRS-1) in a tissue-specific manner. Although the mechanisms underlying the regulation of IRS-1 in the nutrient-rich conditions associated with diabetes and insulin resistance have been well studied, those under nutrient-poor conditions remain unknown. The aim of this study was to investigate how IRS-1 protein levels change depending on the nutritional status of 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were treated with glucose-, amino acid- and serum-free medium for starvation. IRS-1 protein levels were detected by western blot. Autophagy activity was observed by western blot and fluorescence microscopy. The effect of autophagy and p62, an adaptor for selective autophagy, on IRS-1 protein levels under starvation conditions was examined by western blot and immunocytochemistry. We showed that that the levels of IRS-1, but not those of insulin receptor and Akt, decreased when starvation activated autophagy. The inhibition of autophagy by chloroquine or autophagy-related 7 (Atg7) RNA interference counteracted the starvation-induced decrease of IRS-1. Additionally, Atg7 knockdown increased insulin-stimulated phosphorylation of Akt under starvation conditions. Furthermore, p62 co-localized with IRS-1 under starvation conditions, and p62 knockdown counteracted the starvation-induced degradation of IRS-1. Autophagy through p62 plays an important role in regulating IRS-1 protein levels in response to nutritional deficiency. Our findings suggest that autophagy may function as energy depletion-sensing machinery that finely tunes insulin signal transduction. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Proteolysis of serum amyloid A and AA amyloid proteins by cysteine proteases: cathepsin B generates AA amyloid proteins and cathepsin L may prevent their formation

PubMed Central

Rocken, C; Menard, R; Buhling, F; Vockler, S; Raynes, J; Stix, B; Kruger, S; Roessner, A; Kahne, T

2005-01-01

Background: AA amyloidosis develops in patients with chronic inflammatory diseases. The AA amyloid proteins are proteolytic fragments obtained from serum amyloid A (SAA). Previous studies have provided evidence that endosomes or lysosomes might be involved in the processing of SAA, and contribute to the pathology of AA amyloidosis. Objective: To investigate the anatomical distribution of cathepsin (Cath) B and CathL in AA amyloidosis and their ability to process SAA and AA amyloid proteins. Methods and results: CathB and CathL were found immunohistochemically in every patient with AA amyloidosis and displayed a spatial relationship with amyloid in all the cases studied. Both degraded SAA and AA amyloid proteins in vitro. With the help of mass spectrometry 27 fragments were identified after incubation of SAA with CathB, nine of which resembled AA amyloid proteins, and seven fragments after incubation with CathL. CathL did not generate AA amyloid-like peptides. When native human AA amyloid proteins were used as a substrate 26 fragments were identified after incubation with CathB and 18 after incubation with CathL. Conclusion: The two most abundant and ubiquitously expressed lysosomal proteases can cleave SAA and AA amyloid proteins. CathB generates nine AA amyloid-like proteins by its carboxypeptidase activity, whereas CathL may prevent the formation of AA amyloid proteins by endoproteolytic activity within the N-terminal region of SAA. This is particularly interesting, because AA amyloidosis is a systemic disease affecting many organs and tissue types, almost all of which express CathB and CathL. PMID:15897303

A GTPase reaction accompanying the rejection of Leu-tRNA2 by UUU-programmed ribosomes. Proofreading of the codon-anticodon interaction by ribosomes.

PubMed

Thompson, R C; Dix, D B; Gerson, R B; Karim, A M

1981-01-10

The characteristics of a GTPase reaction between poly(U)-programmed ribosomes, EFTu . GTP, and the near-cognate aminoacyl (aa)-tRNA, Leu-tRNA Leu 2, have been studied to assess the role of this reaction in proofreading of the codon-anticodon interaction. The reaction resembles the GTPase reaction with cognate aa-tRNAs and EFTu . GTP in its substrate requirements, in its involving EFTu . GTP . aa-tRNA ternary complexes, and in its requiring a free ribosomal A-site. The noncognate reaction differs from the cognate one in that aa-tRNA becomes stably bound to the ribosomes only 5% of the time; it therefore seems best characterized as an abortive enzymatic binding reaction. The rate of reaction is a significant fraction (4%) of that of the cognate aa-tRNA, indicating that recognition of ternary complexes by ribosomes involves a level of error greater than that of translation as a whole. The rejection of the noncognate aa-tRNA following GTP hydrolysis is therefore a vital step in the translation process and fulfills the criteria set for a proofreading reaction. Leu-tRNA Leu 2 which escapes rejection through proofreading, forms a stable complex with the ribosomal A-site, so it appears that the Leu-tRNA2 which was rejected never reached the A-site and that proofreading precedes full A-site binding.

AaEIN3 Mediates the Downregulation of Artemisinin Biosynthesis by Ethylene Signaling Through Promoting Leaf Senescence in Artemisia annua.

PubMed

Tang, Yueli; Li, Ling; Yan, Tingxiang; Fu, Xueqing; Shi, Pu; Shen, Qian; Sun, Xiaofen; Tang, Kexuan

2018-01-01

Artemisinin is an important drug for malaria treatment, which is exclusively produced in Artemisia annua . It's important to dissect the regulatory mechanism of artemisinin biosynthesis by diverse plant hormones and transcription factors. Our study shows ethylene, a plant hormone which accelerates flower and leaf senescence and fruit ripening, suppressed the expression of genes encoding three key enzymes ADS, DBR2, CYP71AV1, and a positive regulator AaORA involved in artemisinin biosynthesis. Then we isolated the gene encoding ETHYLENE-INSENSITIVE3 (EIN3), a key transcription factor in ethylene signaling pathway, by screening the transcriptome and genome database from Artemisia annua , named AaEIN3 . Overexpressing AaEIN3 suppressed artemisinin biosynthesis, while repressing its expression with RNAi enhanced artemisinin biosynthesis in Artemisia annua , indicating AaEIN3 negatively regulates artemisinin biosynthesis. Further study showed the downregulation of artemisinin biosynthesis by ethylene required the mediation of AaEIN3. AaEIN3 could accelerate leaf senescence, and leaf senescence attenuated the expression of ADS, DBR2, CYP71AV1 , and AaORA that are involved in artemisinin biosynthesis. Collectively, our study demonstrated a negative correlation between ethylene signaling and artemisinin biosynthesis, which is ascribed to AaEIN3-induced senescence process of leaves. Our work provided novel knowledge on the regulatory network of plant hormones for artemisinin metabolic pathway.

AaEIN3 Mediates the Downregulation of Artemisinin Biosynthesis by Ethylene Signaling Through Promoting Leaf Senescence in Artemisia annua

PubMed Central

Tang, Yueli; Li, Ling; Yan, Tingxiang; Fu, Xueqing; Shi, Pu; Shen, Qian; Sun, Xiaofen; Tang, Kexuan

2018-01-01

Artemisinin is an important drug for malaria treatment, which is exclusively produced in Artemisia annua. It’s important to dissect the regulatory mechanism of artemisinin biosynthesis by diverse plant hormones and transcription factors. Our study shows ethylene, a plant hormone which accelerates flower and leaf senescence and fruit ripening, suppressed the expression of genes encoding three key enzymes ADS, DBR2, CYP71AV1, and a positive regulator AaORA involved in artemisinin biosynthesis. Then we isolated the gene encoding ETHYLENE-INSENSITIVE3 (EIN3), a key transcription factor in ethylene signaling pathway, by screening the transcriptome and genome database from Artemisia annua, named AaEIN3. Overexpressing AaEIN3 suppressed artemisinin biosynthesis, while repressing its expression with RNAi enhanced artemisinin biosynthesis in Artemisia annua, indicating AaEIN3 negatively regulates artemisinin biosynthesis. Further study showed the downregulation of artemisinin biosynthesis by ethylene required the mediation of AaEIN3. AaEIN3 could accelerate leaf senescence, and leaf senescence attenuated the expression of ADS, DBR2, CYP71AV1, and AaORA that are involved in artemisinin biosynthesis. Collectively, our study demonstrated a negative correlation between ethylene signaling and artemisinin biosynthesis, which is ascribed to AaEIN3-induced senescence process of leaves. Our work provided novel knowledge on the regulatory network of plant hormones for artemisinin metabolic pathway. PMID:29675029

The Catalytic Domain of Topological Knot tRNA Methyltransferase (TrmH) Discriminates between Substrate tRNA and Nonsubstrate tRNA via an Induced-fit Process*

PubMed Central

Ochi, Anna; Makabe, Koki; Yamagami, Ryota; Hirata, Akira; Sakaguchi, Reiko; Hou, Ya-Ming; Watanabe, Kazunori; Nureki, Osamu; Kuwajima, Kunihiro; Hori, Hiroyuki

2013-01-01

A conserved guanosine at position 18 (G18) in the D-loop of tRNAs is often modified to 2′-O-methylguanosine (Gm). Formation of Gm18 in eubacterial tRNA is catalyzed by tRNA (Gm18) methyltransferase (TrmH). TrmH enzymes can be divided into two types based on their substrate tRNA specificity. Type I TrmH, including Thermus thermophilus TrmH, can modify all tRNA species, whereas type II TrmH, for example Escherichia coli TrmH, modifies only a subset of tRNA species. Our previous crystal study showed that T. thermophilus TrmH is a class IV S-adenosyl-l-methionine-dependent methyltransferase, which maintains a topological knot structure in the catalytic domain. Because TrmH enzymes have short stretches at the N and C termini instead of a clear RNA binding domain, these stretches are believed to be involved in tRNA recognition. In this study, we demonstrate by site-directed mutagenesis that both N- and C-terminal regions function in tRNA binding. However, in vitro and in vivo chimera protein studies, in which four chimeric proteins of type I and II TrmHs were used, demonstrated that the catalytic domain discriminates substrate tRNAs from nonsubstrate tRNAs. Thus, the N- and C-terminal regions do not function in the substrate tRNA discrimination process. Pre-steady state analysis of complex formation between mutant TrmH proteins and tRNA by stopped-flow fluorescence measurement revealed that the C-terminal region works in the initial binding process, in which nonsubstrate tRNA is not excluded, and that structural movement of the motif 2 region of the catalytic domain in an induced-fit process is involved in substrate tRNA discrimination. PMID:23867454

A Bottom-Up Proteomic Approach to Identify Substrate Specificity of Outer-Membrane Protease OmpT.

PubMed

Wood, Sarah E; Sinsinbar, Gaurav; Gudlur, Sushanth; Nallani, Madhavan; Huang, Che-Fan; Liedberg, Bo; Mrksich, Milan

2017-12-22

Identifying peptide substrates that are efficiently cleaved by proteases gives insights into substrate recognition and specificity, guides development of inhibitors, and improves assay sensitivity. Peptide arrays and SAMDI mass spectrometry were used to identify a tetrapeptide substrate exhibiting high activity for the bacterial outer-membrane protease (OmpT). Analysis of protease activity for the preferred residues at the cleavage site (P1, P1') and nearest-neighbor positions (P2, P2') and their positional interdependence revealed FRRV as the optimal peptide with the highest OmpT activity. Substituting FRRV into a fragment of LL37, a natural substrate of OmpT, led to a greater than 400-fold improvement in OmpT catalytic efficiency, with a k cat /K m value of 6.1×10 6  L mol -1  s -1 . Wild-type and mutant OmpT displayed significant differences in their substrate specificities, demonstrating that even modest mutants may not be suitable substitutes for the native enzyme. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Crystal Structure and Substrate Specificity of Drosophila 3,4-Dihydroxyphenylalanine Decarboxylase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Han, Q.; Ding, H; Robinson, H

2010-01-01

3,4-Dihydroxyphenylalanine decarboxylase (DDC), also known as aromatic L-amino acid decarboxylase, catalyzes the decarboxylation of a number of aromatic L-amino acids. Physiologically, DDC is responsible for the production of dopamine and serotonin through the decarboxylation of 3,4-dihydroxyphenylalanine and 5-hydroxytryptophan, respectively. In insects, both dopamine and serotonin serve as classical neurotransmitters, neuromodulators, or neurohormones, and dopamine is also involved in insect cuticle formation, eggshell hardening, and immune responses. In this study, we expressed a typical DDC enzyme from Drosophila melanogaster, critically analyzed its substrate specificity and biochemical properties, determined its crystal structure at 1.75 Angstrom resolution, and evaluated the roles residues T82more » and H192 play in substrate binding and enzyme catalysis through site-directed mutagenesis of the enzyme. Our results establish that this DDC functions exclusively on the production of dopamine and serotonin, with no activity to tyrosine or tryptophan and catalyzes the formation of serotonin more efficiently than dopamine. The crystal structure of Drosophila DDC and the site-directed mutagenesis study of the enzyme demonstrate that T82 is involved in substrate binding and that H192 is used not only for substrate interaction, but for cofactor binding of drDDC as well. Through comparative analysis, the results also provide insight into the structure-function relationship of other insect DDC-like proteins.« less

Cytotoxic 20,24-epoxywithanolides from Physalis angulata.

PubMed

Maldonado, Emma; Hurtado, Norma E; Pérez-Castorena, Ana L; Martínez, Mahinda

2015-12-01

A new withanolide, physangulide B (1), was isolated from calyxes of Physalis angulata. This compound was also present in the aerial parts along with the known physangulide (4), which was isolated as the acetonide 3, and 24,25-epoxywithanolide D (6). Structures of these compounds were determined by analysis of their spectroscopic data, which indicated the presence of a 20,24-epoxy group in both physangulides. The structures of compounds 1 and 6 were confirmed by X-ray analysis of their corresponding acetyl derivatives 2 and 7. The structure of physangulide was originally described as the 22S withanolide 5, now its structure and configuration are revised to 4. Evaluation of the cytotoxic activity of compounds 1-3 against two human cancer cell lines indicated a potent activity of compound 1 and its derivative 2. Copyright © 2015 Elsevier Inc. All rights reserved.

NGC 2024: Far-infrared and radio molecular observations

NASA Technical Reports Server (NTRS)

Thronson, H. A., Jr.; Lada, C. J.; Schwartz, P. R.; Smith, H. A.; Smith, J.; Glaccum, W.; Harper, D. A.; Loewenstein, R. F.

1984-01-01

Far infrared continuum and millimeter wave molecular observations are presented for the infrared and radio source NGC 2024. The measurements are obtained at relatively high angular resolution, enabling a description of the source energetics and mass distribution in greater detail than previously reported. The object appears to be dominated by a dense ridge of material, extended in the north/south direction and centered on the dark lane that is seen in visual photographs. Maps of the source using the high density molecules CS and HCN confirm this picture and allow a description of the core structure and molecular abundances. The radio molecular and infrared observations support the idea that an important exciting star in NGC 2024 has yet to be identified and is centered on the dense ridge about 1' south of the bright mid infrared source IRS 2. The data presented here allows a presentation of a model for the source.

Chemically activated nanodiamonds for aluminum alloy corrosion protection and monitoring

NASA Astrophysics Data System (ADS)

Hannstein, Inga; Adler, Anne-Katrin; Lapina, Victoria; Osipov, Vladimir; Opitz, Jörg; Schreiber, Jürgen; Meyendorf, Norbert

2009-03-01

In the present study, a smart coating for light metal alloys was developed and investigated. Chemically activated nanodiamonds (CANDiT) were electrophoretically deposited onto anodized aluminum alloy AA2024 substrates in order to increase corrosion resistance, enhance bonding properties and establish a means of corrosion monitoring based on the fluorescence behavior of the particles. In order to create stable aqueous CANDiT dispersions suitable for electrophoretic deposition, mechanical milling had to be implemented under specific chemical conditions. The influence of the CANDiT volume fraction and pH of the dispersion on the electrochemical properties of the coated samples was investigated. Linear voltammetry measurements reveal that the chemical characteristics of the CANDiT dispersion have a distinct influence on the quality of the coating. The fluorescence spectra as well as fluorescence excitation spectra of the samples show that corrosion can be easily detected by optical means. Furthermore, an optimization on the basis of "smart" - algorithms for the data processing of a surface analysis by the laser-speckle-method is presented.

49 CFR 178.37 - Specification 3AA and 3AAX seamless steel cylinders.

Code of Federal Regulations, 2011 CFR

2011-10-01

... conform to the following: (1) A DOT-3AA cylinder is a seamless steel cylinder with a water capacity... a seamless steel cylinder with a water capacity of not less than 1,000 pounds and a service pressure... the weight per unit of length of the straight cylindrical portion filled with water and compressed to...

In patients with chronic aplastic anemia, bone marrow-derived MSCs regulate the Treg/Th17 balance by influencing the Notch/RBP-J/FOXP3/RORγt pathway.

PubMed

Li, Hongbo; Wang, Lin; Pang, Yan; Jiang, Zujun; Liu, Zenghui; Xiao, Haowen; Chen, Haijia; Ge, Xiaohu; Lan, Hai; Xiao, Yang

2017-02-14

The standard treatment for aplastic anemia (AA) in young patients is a matched sibling hematopoietic stem cell transplant. Transfusion of a chronic AA patient with allogeneic bone marrow-derived mesenchymal stromal cells (BMMSCs) is currently being developed as a cell-based therapy, and the safety and efficacy of such transfusions are being continuously improved. Nevertheless, the mechanisms by which BMMSCs exert their therapeutic effects remain to be elucidated. In this study, mesenchymal stromal cells (MSCs) obtained from bone marrow donors were concentrated and intravenously injected into 15 chronic AA patients who had been refractory to prior immunosuppressive therapy. We showed that BMMSCs modulate the levels of Th1, Th2, Th17 and Treg cells, as well as their related cytokines in chronic AA patients. Furthermore, the percentages of Th1 and Th17 cells among the H-MSCs decreased significantly, while the percentage Treg cells increased. The Notch/RBP-J/FOXP3/RORγt pathway was involved in modulating the Treg/Th17 balance after MSCs were transfused in vitro. Additionally, the role played by transfused MSCs in regulating the Treg/Th17 balance via the Notch/RBP-J/FOXP3/RORγt pathway was further confirmed in an AA mouse model. In summary, in humans with chronic AA, BMMSCs regulate the Treg/Th17 balance by affecting the Notch/RBP-J/FOXP3/RORγt pathway.

28 CFR 20.24 - State laws on privacy and security.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Local Criminal History Record Information Systems § 20.24 State laws on privacy and security. Where a State originating criminal history record information provides for sealing or purging thereof, nothing...

28 CFR 20.24 - State laws on privacy and security.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Local Criminal History Record Information Systems § 20.24 State laws on privacy and security. Where a State originating criminal history record information provides for sealing or purging thereof, nothing...

28 CFR 20.24 - State laws on privacy and security.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Local Criminal History Record Information Systems § 20.24 State laws on privacy and security. Where a State originating criminal history record information provides for sealing or purging thereof, nothing...

28 CFR 20.24 - State laws on privacy and security.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Local Criminal History Record Information Systems § 20.24 State laws on privacy and security. Where a State originating criminal history record information provides for sealing or purging thereof, nothing...

28 CFR 20.24 - State laws on privacy and security.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Local Criminal History Record Information Systems § 20.24 State laws on privacy and security. Where a State originating criminal history record information provides for sealing or purging thereof, nothing...

Microstructural features of friction stir welded dissimilar Aluminium alloys AA2219-AA7475

NASA Astrophysics Data System (ADS)

Zaman Khan, Noor; Ubaid, Mohammed; Siddiquee, Arshad Noor; Khan, Zahid A.; Al-Ahmari, Abdulrahman; Chen, Xizhang; Haider Abidi, Mustufa

2018-05-01

High strength, good corrosion resistance, light weight make aluminium alloys a material of choice in many industrial sectors like aerospace, marine etc. Problems associated with welding of these alloys by fusion welding processes restricted their use in various industries. Friction stir welding (FSW), a clean solid-state joining process, easily overcomes various difficulties encountered during conventional fusion welding processes. In the present work, the effect of rotational speed (710 rpm, 900 rpm and 1120 rpm) on micro-hardness distribution and microstructure of FSWed dissimilar aluminium alloy joints were analyzed. Plates of AA7475-T761 and AA2219-O having thickness of 2.5 mm were welded by fixing AA7475 on retreating side (RS) and AA2219 on advancing side (AS). Welded joints were characterized by Vickers micro-hardness testing, scanning electron microscopy (SEM) and optical microscopy (OM). Results revealed that rotational speed significantly affects the micro-hardness due to increase in grain size, coarsening and dissolution of strengthening precipitates and re-precipitation. Higher micro-hardness values were observed in stir zone due to grain refinement and re-precipitation. Minimum micro-hardness value was observed at the TMAZ/HAZ of advancing side due to thermal softening.

184AA3: a xenograft model of ER+ breast adenocarcinoma

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hines, William C.; Kuhn, Irene; Thi, Kate

Despite the prevalence and significant morbidity resulting from estrogen receptor positive (ER +) breast adenocarcinomas, there are only a few models of this cancer subtype available for drug development and arguably none for studying etiology. Those models that do exist have questionable clinical relevance. Given our goal of developing luminal models, we focused on six cell lines derived by minimal mutagenesis from normal human breast cells, and asked if any could generate clinically relevant xenografts, which we then extensively characterized. Xenografts of one cell line, 184AA3, consistently formed ER + adenocarcinomas that had a high proliferative rate and other features consistentmore » with “luminal B” intrinsic subtype. Squamous and spindle cell/mesenchymal differentiation was absent, in stark contrast to other cell lines that we examined or others have reported. We explored intratumoral heterogeneity produced by 184AA3 by immunophenotyping xenograft tumors and cultured cells, and characterized marker expression by immunofluorescence and flow cytometry. A CD44 High  subpopulation was discovered, yet their tumor forming ability was far less than CD44 Low  cells. Single cell cloning revealed the phenotypic plasticity of 184AA3, consistent with the intratumoral heterogeneity observed in xenografts. Characterization of ER expression in cultures revealed ER protein and signaling is intact, yet when estrogen was depleted in culture, and in vivo, it did not impact cell or tumor growth, analogous to therapeutically resistant ER +  cancers. In conclusion, this model is appropriate for studies of the etiology of ovarian hormone independent adenocarcinomas, for identification of therapeutic targets, predictive testing, and drug development.« less

184AA3: a xenograft model of ER+ breast adenocarcinoma

DOE PAGES

Hines, William C.; Kuhn, Irene; Thi, Kate; ...

2015-12-12

Despite the prevalence and significant morbidity resulting from estrogen receptor positive (ER +) breast adenocarcinomas, there are only a few models of this cancer subtype available for drug development and arguably none for studying etiology. Those models that do exist have questionable clinical relevance. Given our goal of developing luminal models, we focused on six cell lines derived by minimal mutagenesis from normal human breast cells, and asked if any could generate clinically relevant xenografts, which we then extensively characterized. Xenografts of one cell line, 184AA3, consistently formed ER + adenocarcinomas that had a high proliferative rate and other features consistentmore » with “luminal B” intrinsic subtype. Squamous and spindle cell/mesenchymal differentiation was absent, in stark contrast to other cell lines that we examined or others have reported. We explored intratumoral heterogeneity produced by 184AA3 by immunophenotyping xenograft tumors and cultured cells, and characterized marker expression by immunofluorescence and flow cytometry. A CD44 High  subpopulation was discovered, yet their tumor forming ability was far less than CD44 Low  cells. Single cell cloning revealed the phenotypic plasticity of 184AA3, consistent with the intratumoral heterogeneity observed in xenografts. Characterization of ER expression in cultures revealed ER protein and signaling is intact, yet when estrogen was depleted in culture, and in vivo, it did not impact cell or tumor growth, analogous to therapeutically resistant ER +  cancers. In conclusion, this model is appropriate for studies of the etiology of ovarian hormone independent adenocarcinomas, for identification of therapeutic targets, predictive testing, and drug development.« less

IMAGING THE DISK AND JET OF THE CLASSICAL T TAURI STAR AA TAU

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cox, Andrew W.; Grady, Carol A.; Hammel, Heidi B.

2013-01-01

Previous studies of the classical T Tauri star AA Tau have interpreted the UX-Orionis-like photo-polarimetric variability as being due to a warp in the inner disk caused by an inclined stellar magnetic dipole field. We test that these effects are macroscopically observable in the inclination and alignment of the disk. We use Hubble Space Telescope (HST)/STIS coronagraphic imagery to measure the V magnitude of the star for both STIS coronagraphic observations, compare these data with optical photometry in the literature, and find that, unlike other classical T Tauri stars observed in the same HST program, the disk is most robustlymore » detected in scattered light at stellar optical minimum light. We measure the outer disk radius, 1.''15 {+-} 0.''10, major-axis position angle, and disk inclination and find that the inner disk, as reported in the literature, is both misinclined and misaligned with respect to the outer disk. AA Tau drives a faint jet, detected in both STIS observations and in follow-on Goddard Fabry-Perot imagery, which is also misaligned with respect to the projection of the outer disk minor axis and is poorly collimated near the star, but which can be traced 21'' from the star in data from 2005. The measured outer disk inclination, 71 Degree-Sign {+-} 1 Degree-Sign , is out of the range of inclinations suggested for stars with UX-Orionis-like variability when no grain growth has occurred in the disk. The faintness of the disk, small disk size, and detection of the star despite the high inclination all indicate that the dust disk must have experienced grain growth and settling toward the disk midplane, which we verify by comparing the observed disk with model imagery from the literature.« less

Imaging the Disk and Jet of the Classical T Tauri Star AA Tau

NASA Technical Reports Server (NTRS)

Cox, Andrew W.; Grady, Carol A.; Hammel, Heidi B.; Hornbeck, Jeremy; Russell, Ray W.; Sitko, Michael L.; Woodgate, Bruce E.

2013-01-01

Previous studies of the classical T Tauri star AA Tau have interpreted the UX-Orionis-like photo-polarimetric variability as being due to a warp in the inner disk caused by an inclined stellar magnetic dipole field. We test that these effects are macroscopically observable in the inclination and alignment of the disk. We use Hubble Space Telescope (HST)/STIS coronagraphic imagery to measure the V magnitude of the star for both STIS coronagraphic observations, compare these data with optical photometry in the literature, and find that, unlike other classical T Tauri stars observed in the same HST program, the disk is most robustly detected in scattered light at stellar optical minimum light.We measure the outer disk radius, 1 inch.15 plus-minus 0 inch.10, major-axis position angle, and disk inclination and find that the inner disk, as reported in the literature, is both misinclined and misaligned with respect to the outer disk. AA Tau drives a faint jet, detected in both STIS observations and in follow-on Goddard Fabry-Perot imagery, which is also misaligned with respect to the projection of the outer disk minor axis and is poorly collimated near the star, but which can be traced 21 inches from the star in data from 2005. The measured outer disk inclination, 71deg plus-minus 1deg, is out of the range of inclinations suggested for stars with UX-Orionis-like variability when no grain growth has occurred in the disk. The faintness of the disk, small disk size, and detection of the star despite the high inclination all indicate that the dust disk must have experienced grain growth and settling toward the disk midplane, which we verify by comparing the observed disk with model imagery from the literature.

Mechanical Properties and Microstructural Evolution of Friction-Stir-Welded Thin Sheet Aluminum Alloys

NASA Astrophysics Data System (ADS)

Cerri, Emanuela; Leo, Paola; Wang, Xiang; Embury, J. D.

2011-05-01

Friction stir welding of thin aluminum sheets represents a potential goal for aircraft and automotive industries because of the advantages of using this new technological process. In the current work, the microstructural evolution and mechanical behavior of 6082T6-6082T6, 2024T3-2024T3, and 6082T6-2024T3 thin friction-stir-welded joints were investigated. Uniaxial tensile testing at room temperature, 443 K, 473 K, and 503 K (170 °C, 200 °C, and 230 °C) was used to determine the extent to which these ultra-thin joints can be used and deformed. The tensile stress-strain curves showed a decrease of the flow stress with increasing temperature and decreasing strain rate. The ductility of 6082T6-6082T6 joints generally improved when deformed at warm temperatures. It was almost constant for the 6082T6-2024T3 and reached the higher value in the 2024T3-2024T3 when deformed at 443 K and 473 K (170 °C and 200 °C) when compared with the room temperature value. Tensile specimens fractured in the middle of the weld zone in a ductile mode. The precipitation and growth of S' type phases strengthens 2024T3-2024T3 joints during deformation. In the 6082T6-6082T6, β″ precipitates show some increase in size but give a lower contribution to strength. At 503 K (230 °C), recovery mechanisms (dislocation reorganization inside the deformed grains) are initiated but the temperature was not enough high to produce a homogeneous subgrain structure.

Lessons from Distributing Eclipse Glasses: Planning Ahead for April 2024

NASA Astrophysics Data System (ADS)

Bartlett, Jennifer Lynn; Wilson, Teresa; Chizek Frouard, Malynda R.; Phlips, Alan

2018-01-01

In preparation for the 2017 August 21 total solar eclipse across the continental United States, a multifaceted effort encouraged safe public observation of this spectacular event. However, we experienced mixed results distributing free ISO 12312-2 compliant eclipse glasses.On the positive side, we successfully dispensed several hundred in Virginia through in-school programs about the eclipse. We created a 2017-eclipse information sheet to accompany a safe-viewing handout. To facilitate sending glasses home in student backpacks, we wrapped each pair in a double-sided flyer and sealed the bundle in an individual envelope. We also passed out glasses during evening and weekend activities at a planetarium. Religious, business, and educational groups were all excited to receive them as were co-workers, family, and friends.On the negative side, planetarium staff declined to give eclipse glasses to students without a parent due to safety and liability concerns. Then, a day camp returned 200 pairs less than 72 hours before the event for the same reasons. However, we also received several requests from groups that had waited until too late to be accommodated easily.During the week before the eclipse, demand for eclipse glasses in New York, Michigan, Indiana, Illinois, Wisconsin, Minnesota, South Dakota, Nebraska, and Missouri was less than anticipated. While many people were well prepared, the recalls and reported counterfeiting made others suspicious. Concurrently, vendors were offering their remaining stock for $1–10 each.The experiences of the 2017 total solar eclipse, both good and bad, will not completely fade before preparations for 2024 begin. We look forward enthusiastically to sharing that event with as many people as possible and hope that the overall distribution of eclipse glasses goes more smoothly.We thank the AAS for providing 1,000+ of the eclipse glasses we shared, which were donated to them by Google to promote the Eclipse Megamovie project; Rainbow

50. Representative office, room 2024, second floor, Dwing, building 500, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

50. Representative office, room 2024, second floor, D-wing, building 500, looking west - Offutt Air Force Base, Strategic Air Command Headquarters & Command Center, Headquarters Building, 901 SAC Boulevard, Bellevue, Sarpy County, NE

Assessment of Damage Tolerance Requirements and Analysis. Volume 4. Raw Test Data

DTIC Science & Technology

1986-03-31

T3XX aluminum alloy . Four (4) product forms were selected: MATERIAL PRODUCT FORM THICKNESS (IN) 2024-T3 Sheet 0.090, 0.190 2024-T351 Plate 0.250 2024...T3511 Extruded Tee 0.190 2024-T3511 Extruded Angle 0.250 The material selected to represent bomber/transport/cargo aircraft is 7075- T6XX aluminum alloy ...aluminum alloys , respectively. The raw test data were processed in accordance with ASTM Standard E-8. All the tensile properties, except those marked

Space station integrated wall design and penetration damage control. Task 3: Theoretical analysis of penetration mechanics

NASA Technical Reports Server (NTRS)

Bjorkman, M. D.; Geiger, J. D.; Wilhelm, E. E.

1987-01-01

The efforts to provide a penetration code called PEN4 version 10 is documented for calculation of projectile and target states for the impact of 2024-T3 aluminum, R sub B 90 1018 steel projectiles and icy meteoroids onto 2024-T3 aluminum plates at impact velocities from 0 to 16 km/s. PEN4 determines whether a plate is perforated by calculating the state of fragmentation of projectile and first plate. Depth of penetration into the second to n sup th plate by fragments resulting from first plate perforation is determined by multiple cratering. The results from applications are given.

Transgenic pigeonpea events expressing Cry1Ac and Cry2Aa exhibit resistance to Helicoverpa armigera.

PubMed

Ghosh, Gourab; Ganguly, Shreeparna; Purohit, Arnab; Chaudhuri, Rituparna Kundu; Das, Sampa; Chakraborti, Dipankar

2017-07-01

Independent transgenic pigeonpea events were developed using two cry genes. Transgenic Cry2Aa-pigeonpea was established for the first time. Selected transgenic events demonstrated 100% mortality of Helicoverpa armigera in successive generations. Lepidopteran insect Helicoverpa armigera is the major yield constraint of food legume pigeonpea. The present study was aimed to develop H. armigera-resistant transgenic pigeonpea, selected on the basis of transgene expression and phenotyping. Agrobacterium tumefaciens-mediated transformation of embryonic axis explants of pigeonpea cv UPAS 120 was performed using two separate binary vectors carrying synthetic Bacillus thuringiensis insecticidal crystal protein genes, cry1Ac and cry2Aa. T 0 transformants were selected on the basis of PCR and protein expression profile. T 1 events were exclusively selected on the basis of expression and monogenic character for cry, validated through Western and Southern blot analyses, respectively. Independently transformed 12 Cry1Ac and 11 Cry2Aa single-copy events were developed. The level of Cry-protein expression in T 1 transgenic events was 0.140-0.175% of total soluble protein. Expressed Cry1Ac and Cry2Aa proteins in transgenic pigeonpea exhibited significant weight loss of second-fourth instar larvae of H. armigera and ultimately 80-100% mortality in detached leaf bioassay. Selected Cry-transgenic pigeonpea events, established at T 2 generation, inherited insect-resistant phenotype. Immunohistofluorescence localization in T 3 plants demonstrated constitutive accumulation of Cry1Ac and Cry2Aa in leaf tissues of respective transgenic events. This study is the first report of transgenic pigeonpea development, where stable integration, effective expression and biological activity of two Cry proteins were demonstrated in subsequent three generations (T 0 , T 1, and T 2 ). These studies will contribute to biotechnological breeding programmes of pigeonpea for its genetic improvement.

Microsolvation of the acetanilide cation (AA(+)) in a nonpolar solvent: IR spectra of AA(+)-L(n) clusters (L = He, Ar, N2; n ≤ 10).

PubMed

Schmies, Matthias; Patzer, Alexander; Schütz, Markus; Miyazaki, Mitsuhiko; Fujii, Masaaki; Dopfer, Otto

2014-05-07

Infrared photodissociation (IRPD) spectra of mass-selected cluster ions of acetanilide (N-phenylacetamide), AA(+)-Ln, with the ligands L = He (n = 1-2), Ar (n = 1-7), and N2 (n = 1-10) are recorded in the hydride stretch (amide A, νNH, νCH) and fingerprint (amide I-III) ranges of AA(+) in its (2)A'' ground electronic state. Cold AA(+)-Ln clusters are generated in an electron impact ion source, which predominantly produces the most stable isomer of a given cluster ion. Systematic vibrational frequency shifts of the N-H stretch fundamentals (νNH) provide detailed information about the sequential microsolvation process of AA(+) in a nonpolar (L = He and Ar) and quadrupolar (L = N2) solvent. In the most stable AA(+)-Ln clusters, the first ligand forms a hydrogen bond (H-bond) with the N-H proton of trans-AA(+) (t-AA(+)), whereas further ligands bind weakly to the aromatic ring (π-stacking). There is no experimental evidence for complexes with the less stable cis-AA(+) isomer. Quantum chemical calculations at the M06-2X/aug-cc-pVTZ level confirm the cluster growth sequence derived from the IR spectra. The calculated binding energies of De(H) = 720 and 1227 cm(-1) for H-bonded and De(π) = 585 and 715 cm(-1) for π-bonded Ar and N2 ligands in t-AA(+)-L are consistent with the observed photofragmentation branching ratios of AA(+)-Ln. Comparison between charged and neutral AA((+))-L dimers indicates that ionization switches the preferred ion-ligand binding motif from π-stacking to H-bonding. Electron removal from the HOMO of AA(+) delocalized over both the aromatic ring and the amide group significantly strengthens the C[double bond, length as m-dash]O bond and weakens the N-H bond of the amide group.

Crystal structure of salt-tolerant glutaminase from Micrococcus luteus K-3 in the presence and absence of its product L-glutamate and its activator Tris.

PubMed

Yoshimune, Kazuaki; Shirakihara, Yasuo; Wakayama, Mamoru; Yumoto, Isao

2010-02-01

Glutaminase from Micrococcus luteus K-3 [Micrococcus glutaminase (Mglu); 456 amino acid residues (aa); 48 kDa] is a salt-tolerant enzyme. Our previous study determined the structure of its major 42-kDa fragment. Here, using new crystallization conditions, we determined the structures of the intact enzyme in the presence and absence of its product L-glutamate and its activator Tris, which activates the enzyme by sixfold. With the exception of a 'lid' part (26-29 aa) and a few other short stretches, the structures were all very similar over the entire polypeptide chain. However, the presence of the ligands significantly reduced the length of the disordered regions: 41 aa in the unliganded structure (N), 21 aa for L-glutamate (G), 8 aa for Tris (T) and 6 aa for both L-glutamate and Tris (TG). L-glutamate was identified in both the G and TG structures, whereas Tris was only identified in the TG structure. Comparison of the glutamate-binding site between Mglu and salt-labile glutaminase (YbgJ) from Bacillus subtilis showed significantly smaller structural changes of the protein part in Mglu. A comparison of the substrate-binding pocket of Mglu, which is highly specific for L-glutamine, with that of Erwinia carotovora asparaginase, which has substrates other than L-glutamine, shows that Mglu has a larger substrate-binding pocket that prevents the binding of L-asparagine with proper interactions.

Nanofiber Alignment Regulates NIH3T3 Cell Orientation and Cytoskeletal Gene Expression on Electrospun PCL+Gelatin Nanofibers.

PubMed

Fee, Timothy; Surianarayanan, Swetha; Downs, Crawford; Zhou, Yong; Berry, Joel

2016-01-01

To examine the influence of substrate topology on the behavior of fibroblasts, tissue engineering scaffolds were electrospun from polycaprolactone (PCL) and a blend of PCL and gelatin (PCL+Gel) to produce matrices with both random and aligned nanofibrous orientations. The addition of gelatin to the scaffold was shown to increase the hydrophilicity of the PCL matrix and to increase the proliferation of NIH3T3 cells compared to scaffolds of PCL alone. The orientation of nanofibers within the matrix did not have an effect on the proliferation of adherent cells, but cells on aligned substrates were shown to elongate and align parallel to the direction of substrate fiber alignment. A microarray of cyotoskeleton regulators was probed to examine differences in gene expression between cells grown on an aligned and randomly oriented substrates. It was found that transcriptional expression of eight genes was statistically different between the two conditions, with all of them being upregulated in the aligned condition. The proteins encoded by these genes are linked to production and polymerization of actin microfilaments, as well as focal adhesion assembly. Taken together, the data indicates NIH3T3 fibroblasts on aligned substrates align themselves parallel with their substrate and increase production of actin and focal adhesion related genes.

Dynamic Response of AA2519 Aluminum Alloy under High Strain Rates

NASA Astrophysics Data System (ADS)

Olasumboye, Adewale Taiwo

Like others in the AA2000 series, AA2519 is a heat-treatable Al-Cu alloy. Its excellent ballistic properties and stress corrosion cracking resistance, combined with other properties, qualify it as a prime candidate for armored vehicle and aircraft applications. However, available data on its high strain-rate response remains limited. In this study, AA2519 aluminum alloy was investigated in three different temper conditions: T4, T6, and T8, to determine the effects of heat treatment on the microstructure and dynamic deformation behavior of the material at high strain rates ranging within 1000 ≤ epsilon ≤ 4000 s-1. Split Hopkinson pressure bar integrated with digital image correlation system was used for mechanical response characterization. Optical microscopy and scanning electron microscopy were used to assess the microstructure of the material after following standard metallographic specimen preparation techniques. Results showed heterogeneous deformation in the three temper conditions. It was observed that dynamic behavior in each condition was dependent on strength properties due to the aging type controlling the strengthening precipitates produced and initial microstructure. At 1500 s -1, AA2519-T6 exhibited peak dynamic yield strength and flow stress of 509 and 667 MPa respectively, which are comparable with what were observed in T8 condition at higher rate of 3500 s-1 but AA2519-T4 showed the least strength and flow stress properties. Early stress collapse, dynamic strain aging, and higher susceptibility to shear band formation and fracture were observed in the T6 condition within the selected range of high strain rates. The alloy's general mode of damage evolution was by dispersoid particle nucleation, shearing and cracking.

The Inhibitory Effect of Non-Substrate and Substrate DNA on the Ligation and Self-Adenylylation Reactions Catalyzed by T4 DNA Ligase.

PubMed

Bauer, Robert J; Evans, Thomas C; Lohman, Gregory J S

2016-01-01

DNA ligases are essential both to in vivo replication, repair and recombination processes, and in vitro molecular biology protocols. Prior characterization of DNA ligases through gel shift assays has shown the presence of a nick site to be essential for tight binding between the enzyme and its dsDNA substrate, with no interaction evident on dsDNA lacking a nick. In the current study, we observed a significant substrate inhibition effect, as well as the inhibition of both the self-adenylylation and nick-sealing steps of T4 DNA ligase by non-nicked, non-substrate dsDNA. Inhibition by non-substrate DNA was dependent only on the total DNA concentration rather than the structure; with 1 μg/mL of 40-mers, 75-mers, or circular plasmid DNA all inhibiting ligation equally. A >15-fold reduction in T4 DNA ligase self-adenylylation rate when in the presence of high non-nicked dsDNA concentrations was observed. Finally, EMSAs were utilized to demonstrate that non-substrate dsDNA can compete with nicked dsDNA substrates for enzyme binding. Based upon these data, we hypothesize the inhibition of T4 DNA ligase by non-nicked dsDNA is direct evidence for a two-step nick-binding mechanism, with an initial, nick-independent, transient dsDNA-binding event preceding a transition to a stable binding complex in the presence of a nick site.

The Inhibitory Effect of Non-Substrate and Substrate DNA on the Ligation and Self-Adenylylation Reactions Catalyzed by T4 DNA Ligase

PubMed Central

Bauer, Robert J.; Evans, Thomas C.; Lohman, Gregory J. S.

2016-01-01

DNA ligases are essential both to in vivo replication, repair and recombination processes, and in vitro molecular biology protocols. Prior characterization of DNA ligases through gel shift assays has shown the presence of a nick site to be essential for tight binding between the enzyme and its dsDNA substrate, with no interaction evident on dsDNA lacking a nick. In the current study, we observed a significant substrate inhibition effect, as well as the inhibition of both the self-adenylylation and nick-sealing steps of T4 DNA ligase by non-nicked, non-substrate dsDNA. Inhibition by non-substrate DNA was dependent only on the total DNA concentration rather than the structure; with 1 μg/mL of 40-mers, 75-mers, or circular plasmid DNA all inhibiting ligation equally. A >15-fold reduction in T4 DNA ligase self-adenylylation rate when in the presence of high non-nicked dsDNA concentrations was observed. Finally, EMSAs were utilized to demonstrate that non-substrate dsDNA can compete with nicked dsDNA substrates for enzyme binding. Based upon these data, we hypothesize the inhibition of T4 DNA ligase by non-nicked dsDNA is direct evidence for a two-step nick-binding mechanism, with an initial, nick-independent, transient dsDNA-binding event preceding a transition to a stable binding complex in the presence of a nick site. PMID:26954034

Learning from oligosaccharide soaks of crystals of an AA13 lytic polysaccharide monooxygenase: crystal packing, ligand binding and active-site disorder.

PubMed

Frandsen, Kristian E H; Poulsen, Jens Christian Navarro; Tovborg, Morten; Johansen, Katja S; Lo Leggio, Leila

2017-01-01

Lytic polysaccharide monooxygenases (LPMOs) are a class of copper-dependent enzymes discovered within the last ten years. They oxidatively cleave polysaccharides (chitin, lignocellulose, hemicellulose and starch-derived), presumably making recalcitrant substrates accessible to glycoside hydrolases. Recently, the first crystal structure of an LPMO-substrate complex was reported, giving insights into the interaction of LPMOs with β-linked substrates (Frandsen et al., 2016). The LPMOs acting on α-linked glycosidic bonds (family AA13) display binding surfaces that are quite different from those of LPMOs that act on β-linked glycosidic bonds (families AA9-AA11), as revealed from the first determined structure (Lo Leggio et al., 2015), and thus presumably the AA13s interact with their substrate in a distinct fashion. Here, several new structures of the same AA13 enzyme, Aspergillus oryzae AA13, are presented. Crystals obtained in the presence of high zinc-ion concentrations were used, as they can be obtained more reproducibly than those used to refine the deposited copper-containing structure. One structure with an ordered zinc-bound active site was solved at 1.65 Å resolution, and three structures from crystals soaked with maltooligosaccharides in solutions devoid of zinc ions were solved at resolutions of up to 1.10 Å. Despite similar unit-cell parameters, small rearrangements in the crystal packing occur when the crystals are depleted of zinc ions, resulting in a more occluded substrate-binding surface. In two of the three structures maltooligosaccharide ligands are bound, but not at the active site. Two of the structures presented show a His-ligand conformation that is incompatible with metal-ion binding. In one of these structures this conformation is the principal one (80% occupancy), giving a rare atomic resolution view of a substantially misfolded enzyme that is presumably rendered inactive.

Enhanced osteogenic differentiation of MC3T3-E1 cells on grid-topographic surface and evidence for involvement of YAP mediator.

PubMed

Zhang, Yingying; Gong, He; Sun, Yan; Huang, Yan; Fan, Yubo

2016-05-01

Numerous studies have shown that surface topography can promote cell-substrate associations and deeply influence cell fate. The intracellular mechanism or how micro- or nano-patterned extracellular signal is ultimately linked to activity of nuclear transcription factors remains unknown. It has been reported that Yes-associated protein (YAP) can respond to extracellular matrix microenvironment signals, thus regulates stem cell differentiation process. We propose that YAP may play a role in mediating the topography induced cell differentiation. To this end, we fabricated polydimethylsiloxane (PDMS) micropatterns with grid topology (GT) (3 μm pattern width, 2 μm pattern interval length, 7 μm pattern height); nonpatterned PDMS substrates were used as the planar controls. The MC3T3-E1 cells were then cultured on these surfaces, respectively, in osteogenic inducing medium. Cell differentiation in terms of osteogenesis related gene expression, protein levels, alkaline phosphatase activity and extracellular matrix mineralization was assessed. It was shown that the cells on GT surfaces had stronger osteogenesis capacity. In addition, expression level of YAP was increased when MC3T3-E1 cells grew on GT substrates, which was similar to the levels of osteogenic differentiation markers. It was also shown that YAP knockdown attenuated GT substrates-induced MC3T3-E1 differentiation, which reduced the osteogenic differentiation effect of the GT substrates. Collectively, our findings indicate that GT substrates-induced MC3T3-E1 differentiation may be associated with YAP. This paper provides new target points for transcriptional mechanism research of microenvironment induced cell differentiation and a useful approach to obtain more biofunctionalization scaffolds for tissue engineering. © 2016 Wiley Periodicals, Inc.

mhpT encodes an active transporter involved in 3-(3-hydroxyphenyl)propionate catabolism by Escherichia coli K-12.

PubMed

Xu, Ying; Chen, Bing; Chao, Hongjun; Zhou, Ning-Yi

2013-10-01

Escherichia coli K-12 utilizes 3-(3-hydroxyphenyl)propionate (3HPP) as a sole carbon and energy source. Among the genes in its catabolic cluster in the genome, mhpT was proposed to encode a hypothetical transporter. Since no transporter for 3HPP uptake has been identified, we investigated whether MhpT is responsible for 3HPP uptake. MhpT fused with green fluorescent protein was found to be located at the periphery of cells by confocal microscopy, consistent with localization to the cytoplasmic membrane. Gene knockout and complementation studies clearly indicated that mhpT is essential for 3HPP catabolism in E. coli K-12 W3110 at pH 8.2. Uptake assays with (14)C-labeled substrates demonstrated that strain W3110 and strain W3110ΔmhpT containing recombinant MhpT specifically transported 3HPP but not benzoate, 3-hydroxybenzoate, or gentisate into cells. Energy dependence assays suggested that MhpT-mediated 3HPP transport was driven by the proton motive force. The change of Ala-272 of MhpT to a histidine, surprisingly, resulted in enhanced transport activity, and strain W3110ΔmhpT containing the MhpT A272H mutation had a slightly higher growth rate than the wild-type strain at pH 8.2. Hence, we demonstrated that MhpT is a specific 3HPP transporter and vital for E. coli K-12 W3110 growth on this substrate under basic conditions.

Effect of Weld Tool Geometry on Friction Stir Welded AA2219-T87 Properties

NASA Technical Reports Server (NTRS)

Querin, Joseph A.; Schneider, Judy A.

2008-01-01

In this study, flat panels of AA2219-T87 were friction stir welded (FSWed) using weld tools with tapered pins The three pin geometries of the weld tools included: 0 (straight cylinder), 30 , and 60 angles on the frustum. For each weld tool geometry, the FSW process parameters were optimized to eliminate defects. A constant heat input was maintained while varying the process parameters of spindle rpm and travel speed. This provided a constant heat input for each FSW weld panel while altering the hot working conditions imparted to the workpiece. The resulting mechanical properties were evaluated from tensile test results of the FSW joint.

Identification of GAD65 AA 114-122 reactive 'memory-like' NK cells in newly diagnosed Type 1 diabetic patients by HLA-class I pentamers.

PubMed

Perri, Valentina; Gianchecchi, Elena; Cifaldi, Loredana; Pellegrino, Marsha; Giorda, Ezio; Andreani, Marco; Cappa, Marco; Fierabracci, Alessandra

2017-01-01

Type 1 diabetes is an autoimmune disease, in which pancreatic β cells are destroyed by autoreactive T cells in genetically predisposed individuals. Serum beta cell autoantibody specificities have represented the mainstay for classifying diabetes as autoimmune-mediated and for stratifying risk in first-degree relatives. In recent years, approaches were attempted to solve the difficult issue of detecting rare antigen-specific autoreactive T cells and their significance to etiopathogenesis such as the use of the MHC multimer technology. This tool allowed the specific detection of increased percentages of GAD65 autoreactive T cells by means of HLA A*02:01 GAD65 AA 114-122 pentamers in newly diagnosed diabetics. Here we provide evidence that GAD65 AA 114-122 pentamers can depict a GAD65 AA114-122 peptide expandable population of functionally and phenotypically skewed, preliminary characterized CD3-CD8dullCD56+ 'memory-like' NK cells in PBMC of newly diagnosed diabetics. Our data suggest that the NK cell subset could bind the HLA class I GAD65 AA 114-122 pentamer through ILT2 inhibitory receptor. CD107a expression revealed increased degranulation of CD3-CD8dullCD56+ NK cells in GAD65 AA 114-122 and FLU peptide expanded peripheral blood mononuclear cells of diabetics following GAD65 AA 114-122 peptide HLA A*02:01 presentation in respect to the unpulsed condition. CD107a expression was enriched in ILT2 positive NK cells. As opposite to basal conditions where similar percentages of CD3-CD56+ILT2+ cells were detected in diabetics and controls, CD3-CD56+CD107a+ and CD3-CD56+ILT2+CD107a+ cells were significantly increased in T1D PBMC either GAD65 AA 114-122 or FLU peptides stimulated after co-culture with GAD65 AA 114-122 pulsed APCs. As control, healthy donor NK cells showed similar degranulation against both GAD65 AA 114-122 pulsed and unpulsed APCs. The pathogenetic significance of the CD3-CD8dullCD56+ 'memory-like NK cell subset' with increased response upon secondary

Thyronamines are isozyme-specific substrates of deiodinases.

PubMed

Piehl, S; Heberer, T; Balizs, G; Scanlan, T S; Smits, R; Koksch, B; Köhrle, J

2008-06-01

3-Iodothyronamine (3-T 1 AM) and thyronamine (T AM) are novel endogenous signaling molecules that exhibit great structural similarity to thyroid hormones but apparently antagonize classical thyroid hormone (T(3)) actions. Their proposed biosynthesis from thyroid hormones would require decarboxylation and more or less extensive deiodination. Deiodinases (Dio1, Dio2, and Dio3) catalyze the removal of iodine from their substrates. Because a role of deiodinases in thyronamine biosynthesis requires their ability to accept thyronamines as substrates, we investigated whether thyronamines are converted by deiodinases. Thyronamines were incubated with isozyme-specific deiodinase preparations. Deiodination products were analyzed using a newly established method applying liquid chromatography and tandem mass spectrometry (LC-MS/MS). Phenolic ring deiodinations of 3,3',5'-triiodothyronamine (rT3AM), 3',5'-diiodothyronamine (3',5'-T2AM), and 3,3'-diiodothyronamine (3,3'-T2AM) as well as tyrosyl ring deiodinations of 3,5,3'-triiodothyronamine (T3AM) and 3,5-diiodothyronamine (3,5-T2AM) were observed with Dio1. These reactions were completely inhibited by the Dio1-specific inhibitor 6n-propyl-2-thiouracil (PTU). Dio2 containing preparations also deiodinated rT(3)AM and 3',5'-T2AM at the phenolic rings but in a PTU-insensitive fashion. All thyronamines with tyrosyl ring iodine atoms were 5(3)-deiodinated by Dio3-containing preparations. In functional competition assays, the newly identified thyronamine substrates inhibited an established iodothyronine deiodination reaction. By contrast, thyronamines that had been excluded as deiodinase substrates in LC-MS/MS experiments failed to show any effect in the competition assays, thus verifying the former results. These data support a role for deiodinases in thyronamine biosynthesis and contribute to confining the biosynthetic pathways for 3-T 1 AM and T 0 AM.

A flattened cloud core in NGC 2024

NASA Technical Reports Server (NTRS)

Ho, Paul T. P.; Peng, Yun-Lou; Torrelles, Jose M.; Gomez, Jose F.; Rodriguez, Luis F.; Canto, Jorge

1993-01-01

The (J, K) (1, 1) and (2, 2) NH3 lines were mapped toward a molecular cloud core in NGC 2024 using the VLA in its C/D-configuration. This region is associated with one of the most highly collimated molecular outflows. We find that the molecular condensations associated with the far-infrared sources FIR 5, FIR 6, and FIR 7 have kinetic temperatures of about 40 K. We also find line broadening toward FIR 6 and FIR 7. This suggests that these condensations may not be protostars heated by gravitational energy released during collapse but that they have an internal heating source. A flattened structure of ammonia emission is found extending parallel to the unipolar CO outflow structure, but displaced systematically to the east. If the NH3 emission traces the denser gas environment, there is no evidence that a dense gas structure is confining the molecular outflow. Instead, the location of the high-velocity outflow along the surface of the NH3 structure suggests that a wind is sweeping material from the surface of this elongated cloud core.

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

PubMed Central

Jomaa, Walid; Songmene, Victor; Bocher, Philippe

2014-01-01

The surface finish was extensively studied in usual machining processes (turning, milling, and drilling). For these processes, the surface finish is strongly influenced by the cutting feed and the tool nose radius. However, a basic understanding of tool/surface finish interaction and residual stress generation has been lacking. This paper aims to investigate the surface finish and residual stresses under the orthogonal cutting since it can provide this information by avoiding the effect of the tool nose radius. The orthogonal machining of AA7075-T651 alloy through a series of cutting experiments was performed under dry conditions. Surface finish was studied using height and amplitude distribution roughness parameters. SEM and EDS were used to analyze surface damage and built-up edge (BUE) formation. An analysis of the surface topography showed that the surface roughness was sensitive to changes in cutting parameters. It was found that the formation of BUE and the interaction between the tool edge and the iron-rich intermetallic particles play a determinant role in controlling the surface finish during dry orthogonal machining of the AA7075-T651 alloy. Hoop stress was predominantly compressive on the surface and tended to be tensile with increased cutting speed. The reverse occurred for the surface axial stress. The smaller the cutting feed, the greater is the effect of cutting speed on both axial and hoop stresses. By controlling the cutting speed and feed, it is possible to generate a benchmark residual stress state and good surface finish using dry machining. PMID:28788534

Effective Management of Hexavalent Chromium (Cr+6) in DoD Organic and Inorganic Coatings Operations

DTIC Science & Technology

2014-11-19

stropki@battelle.org 614-424-5414 ASETSDefense 2014 November 19, 2014 Report Documentation Page Form ApprovedOMB No. 0704-0188 Public reporting...19a. NAME OF RESPONSIBLE PERSON a. REPORT unclassified b. ABSTRACT unclassified c. THIS PAGE unclassified Standard Form 298 (Rev. 8-98...Determine impacts of HCR on select aerospace substrates - ferrous (1020 CS and 4340 HSS ), non-ferrous (Al 2024-T3, Al7075-T6, and Mg AZ31B-H24

Negative Affect, Relapse, and Alcoholics Anonymous (AA): Does AA Work by Reducing Anger?*

PubMed Central

Kelly, John F.; Stout, Robert L.; Tonigan, J. Scott; Magill, Molly; Pagano, Maria E.

2010-01-01

Objective: Anger and other indices of negative affect have been implicated in a stress-induced pathway to relapse. The Alcoholics Anonymous (AA) literature states that reduction of anger is critical to recovery, yet this proposed mechanism has rarely been investigated. Using lagged, controlled hierarchical linear modeling analyses, this study investigated whether AA attendance mobilized changes in anger and whether such changes explained AA-related benefit. Method: Alcohol-dependent adults (N = 1,706) receiving treatment as part of a clinical trial were assessed at intake and at 3, 6, 9, 12, and 15 months. Results: Findings revealed substantially elevated levels of anger compared with the general population (98th percentile) that decreased over 15-month follow-up but remained high (89th percentile). AA attendance was associated with better drinking outcomes, and higher levels of anger were associated with heavier drinking. However, AA attendance was unrelated to changes in anger. Conclusions: Although support was not found for anger as a mediator, there was strong convergence between AA's explicit emphasis on anger and the present findings: Anger appears to be a serious, enduring problem related to relapse and heavy alcohol consumption. Methodological factors may have contributed to the lack of association between AA and anger, but results suggest that AA attendance alone may be insufficient to alleviate the suffering and alcohol-related risks specifically associated with anger. PMID:20409438

Transgenic cotton expressing Cry10Aa toxin confers high resistance to the cotton boll weevil.

PubMed

Ribeiro, Thuanne Pires; Arraes, Fabricio Barbosa Monteiro; Lourenço-Tessutti, Isabela Tristan; Silva, Marilia Santos; Lisei-de-Sá, Maria Eugênia; Lucena, Wagner Alexandre; Macedo, Leonardo Lima Pepino; Lima, Janaina Nascimento; Santos Amorim, Regina Maria; Artico, Sinara; Alves-Ferreira, Márcio; Mattar Silva, Maria Cristina; Grossi-de-Sa, Maria Fatima

2017-08-01

Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation. Quantitative PCR (qPCR) assays revealed high transcription levels of cry10Aa in both T 0 GM cotton leaf and flower bud tissues. Southern blot and qPCR-based 2 -ΔΔCt analyses revealed that T 0 GM plants had either one or two transgene copies. Quantitative and qualitative analyses of Cry10Aa protein expression showed variable protein expression levels in both flower buds and leaves tissues of T 0 GM cotton plants, ranging from approximately 3.0 to 14.0 μg g -1 fresh tissue. CBW susceptibility bioassays, performed by feeding adults and larvae with T 0 GM cotton leaves and flower buds, respectively, demonstrated a significant entomotoxic effect and a high level of CBW mortality (up to 100%). Molecular analysis revealed that transgene stability and entomotoxic effect to CBW were maintained in T 1 generation as the Cry10Aa toxin expression levels remained high in both tissues, ranging from 4.05 to 19.57 μg g -1 fresh tissue, and the CBW mortality rate remained around 100%. In conclusion, these Cry10Aa GM cotton plants represent a great advance in the control of the devastating CBW insect pest and can substantially impact cotton agribusiness. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

Effects of Different Filler Metals on the Mechanical Behaviors of GTA Welded AA7A52(T6)

NASA Astrophysics Data System (ADS)

Shu, Fengyuan; Lv, Yaohui; Liu, Yuxin; Lin, Jianjun; Sun, Zhe; Xu, Binshi; He, Peng

2014-06-01

ER4043, ER5356, and AA7A52 on behalf of the Al-Si, Al-Mg, and Al-Zn-Mg-based welding material, respectively, were chosen as the filler metal to weld AA7A52(T6) plates by GTAW. The variance in mechanical performances of the joints caused by the various filler materials was investigated with reference to the SEM and EDS test results for the weld seam and the fracture surface. Failure was found in the seam for all the welded joints. With regard to the joint obtained with ER4043 welding wire, the total elongation was limited by the brittle intergranular compound Mg2Si of which Mg was introduced by convection mass transfer. As for the other two welds, the content ratio of Zn and Mg was found to play the dominant role in deciding the mechanical properties of the intergranular Mg-Zn compounds which were responsible for the tensile behavior of the joints. The content ratio (wt.%) of beyond 2:1 gave birth to the strengthening phase MgZn2 leading to a ductile fracture. Cr in the seam obtained with AA7A52 filler metal was found to enhance the strength of the joint through isolated particles.

Increased toxicity of Bacillus thuringiensis Cry3Aa against Crioceris quatuordecimpunctata, Phaedon brassicae and Colaphellus bowringi by a Tenebrio molitor cadherin fragment.

PubMed

Gao, Yulin; Jurat-Fuentes, Juan Luis; Oppert, Brenda; Fabrick, Jeffrey A; Liu, Chenxi; Gao, Jianhua; Lei, Zhongren

2011-09-01

Biopesticides containing Cry insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are effective against many lepidopteran pests, but there is a lack of Bt-based pesticides for efficient control of important coleopteran pests. Based on the reported increase in Bt toxin oligomerization by a polypeptide from the Cry3Aa receptor cadherin in Tenebrio molitor (Coleoptera: Tenebrionidae), it was hypothesized that this cadherin peptide, rTmCad1p, would enhance Cry3Aa toxicity towards coleopteran larvae. To test this hypothesis, the relative toxicity of Cry3Aa, with or without rTmCad1p, against damaging chrysomelid vegetable pests of China was evaluated. Cry3Aa toxicity was evaluated in the spotted asparagus beetle (Crioceris quatuordecimpunctata), cabbage leaf beetle (Colaphellus bowringi) and daikon leaf beetle (Phaedon brassicae). To assess the effect of rTmCad1p on Cry3Aa toxicity, neonate larvae were fed Cry3Aa toxin alone or in combination with increasing amounts of rTmCad1p. The data demonstrated that Cry3Aa toxicity was significantly increased in all three vegetable pests, resulting in as much as a 15.3-fold increase in larval mortality. The application of rTmCad1p to enhance Cry3Aa insecticidal activity has potential for use in increasing range and activity levels against coleopteran pests displaying low susceptibility to Bt-based biopesticides. Copyright © 2011 Society of Chemical Industry.

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.

Experimental Investigation on Friction Stir Welding of Cryorolled AA2219 Aluminum Alloy Joints

NASA Astrophysics Data System (ADS)

Babu, K. Kamal; Panneerselvam, K.; Sathiya, P.; Haq, A. Noorul; Sundarrajan, S.; Mastanaiah, P.; Murthy, C. V. Srinivasa

2017-07-01

In this paper, experimental investigation on cryorolled aluminum AA2219-T87 plate by using friction stir welding (FSW) process is carried out. AA2219-T87 plates with a size of 200×100×22.4 mm were rolled and reduced to 12.2mm thickness (more than 45% of reduction in total thickness of the base material) at cryogenic temperature (operating temperature range -90--30∘C). The cryorolled (CR) plates have reduced grain size, improved hardness and increased corrosion resistance property compared with the uncryorolled AA2219-T87 plates. FSW joints of cryorolled AA2219-T87 plates were prepared using cylindrical threaded FSW tool pin profile. Mechanical and metallurgical behaviors of friction stir welded joints were analyzed and the effects of the FSW process parameters are discussed in this paper. The variation of microhardness in the FSW joint regions were correlated with the microstructure of FSW joints. Cryorolled plate and FSW joints were tested for corrosion resistance using potentiodynamic polarization test. FSW joints shows better result during the corrosion resistance analysis compared to base AA2219-T87. The X-ray diffraction (XRD) test results showed that fine α-Al grains with eutectic phase (Al2Cu) were present in the weld nugget (WN). The large clusters of strengthening precipitates were reduced in size and merged with the weld nugget portion.

Thyronamines Are Isozyme-Specific Substrates of Deiodinases

PubMed Central

Piehl, S.; Heberer, T.; Balizs, G.; Scanlan, T. S.; Smits, R.; Koksch, B.; Köhrle, J.

2008-01-01

3-Iodothyronamine (3-T1AM) and thyronamine (T0AM) are novel endogenous signaling molecules that exhibit great structural similarity to thyroid hormones but apparently antagonize classical thyroid hormone (T3) actions. Their proposed biosynthesis from thyroid hormones would require decarboxylation and more or less extensive deiodination. Deiodinases (Dio1, Dio2, and Dio3) catalyze the removal of iodine from their substrates. Because a role of deiodinases in thyronamine biosynthesis requires their ability to accept thyronamines as substrates, we investigated whether thyronamines are converted by deiodinases. Thyronamines were incubated with isozyme-specific deiodinase preparations. Deiodination products were analyzed using a newly established method applying liquid chromatography and tandem mass spectrometry (LC-MS/MS). Phenolic ring deiodinations of 3,3′,5′-triiodothyronamine (rT3AM), 3′,5′-diiodothyronamine (3′,5′-T2AM), and 3,3′-diiodothyronamine (3,3′-T2AM) as well as tyrosyl ring deiodinations of 3,5,3′-triiodothyronamine (T3AM) and 3,5-diiodothyronamine (3,5-T2AM) were observed with Dio1. These reactions were completely inhibited by the Dio1-specific inhibitor 6n-propyl-2-thiouracil (PTU). Dio2 containing preparations also deiodinated rT3AM and 3′,5′-T2AM at the phenolic rings but in a PTU-insensitive fashion. All thyronamines with tyrosyl ring iodine atoms were 5(3)-deiodinated by Dio3-containing preparations. In functional competition assays, the newly identified thyronamine substrates inhibited an established iodothyronine deiodination reaction. By contrast, thyronamines that had been excluded as deiodinase substrates in LC-MS/MS experiments failed to show any effect in the competition assays, thus verifying the former results. These data support a role for deiodinases in thyronamine biosynthesis and contribute to confining the biosynthetic pathways for 3-T1AM and T0AM. PMID:18339710

What Are the bona fide GSK3 Substrates?

PubMed

Sutherland, Calum

2011-01-01

Nearly 100 proteins are proposed to be substrates for GSK3, suggesting that this enzyme is a fundamental regulator of almost every process in the cell, in every tissue in the body. However, it is not certain how many of these proposed substrates are regulated by GSK3 in vivo. Clearly, the identification of the physiological functions of GSK3 will be greatly aided by the identification of its bona fide substrates, and the development of GSK3 as a therapeutic target will be highly influenced by this range of actions, hence the need to accurately establish true GSK3 substrates in cells. In this paper the evidence that proposed GSK3 substrates are likely to be physiological targets is assessed, highlighting the key cellular processes that could be modulated by GSK3 activity and inhibition.

Effect of the Microstructure on Diffusion Bonded AA5083, AA6082 and AA7075 Aluminium Alloys

NASA Astrophysics Data System (ADS)

Venugopal, S.; Mahendran, G.

2018-05-01

Rolled plates of aluminium alloys AA5083, AA6082 and AA7075 of 5 mm thickness are joined by diffusion bonding at varied parameters. The microstructure evolution of AA5083, AA6082 and AA7075 aluminium alloys is characterized by Transmission Electron Microscopy (TEM). Metallurgical investigations and mechanical tests are also performed to correlate the results of the TEM investigations with the mechanical properties of the produced diffusion bonded joints. It is observed that the bonding and shear strength of the alloys increase with the increase in bonding temperature, due to the diffusion of micro-constituents in the interface. High temperature enhances the uniform distribution of secondary phase particles and reduces pore formation/defects in the bonded joints.

Sub-Cellular Localization and Complex Formation by Aminoacyl-tRNA Synthetases in Cyanobacteria: Evidence for Interaction of Membrane-Anchored ValRS with ATP Synthase.

PubMed

Santamaría-Gómez, Javier; Ochoa de Alda, Jesús A G; Olmedo-Verd, Elvira; Bru-Martínez, Roque; Luque, Ignacio

2016-01-01

tRNAs are charged with cognate amino acids by aminoacyl-tRNA synthetases (aaRSs) and subsequently delivered to the ribosome to be used as substrates for gene translation. Whether aminoacyl-tRNAs are channeled to the ribosome by transit within translational complexes that avoid their diffusion in the cytoplasm is a matter of intense investigation in organisms of the three domains of life. In the cyanobacterium Anabaena sp. PCC 7120, the valyl-tRNA synthetase (ValRS) is anchored to thylakoid membranes by means of the CAAD domain. We have investigated whether in this organism ValRS could act as a hub for the nucleation of a translational complex by attracting other aaRSs to the membranes. Out of the 20 aaRSs, only ValRS was found to localize in thylakoid membranes whereas the other enzymes occupied the soluble portion of the cytoplasm. To investigate the basis for this asymmetric distribution of aaRSs, a global search for proteins interacting with the 20 aaRSs was conducted. The interaction between ValRS and the FoF1 ATP synthase complex here reported is of utmost interest and suggests a functional link between elements of the gene translation and energy production machineries.

High speed machinability of the aerospace alloy AA7075 T6 under different cooling conditions

NASA Astrophysics Data System (ADS)

Imbrogno, Stano; Rinaldi, Sergio; Suarez, Asier Gurruchaga; Arrazola, Pedro J.; Umbrello, Domenico

2018-05-01

This paper describes the results of an experimental investigation aimed to st udy the machinability of AA7075 T6 (160 HV) for aerospace industry at high cutting speeds. The paper investigates the effects of different lubri-cooling strategies (cryogenic, M QL and dry) during high speed turning process on cutting forces, tool wear, chip morphology and cutting temperatures. The cutting speeds selected were 1000m/min, 1250m/min and 1500 m/min, while the feed rate values used were 0.1mm/rev and 0.3 mm/rev. The results of cryogenic and M QL application is compared with dry application. It was found that the cryogenic and M QL lubri-cooling techniques could represent a functional alternative to the common dry cutting application in order to implement a more effect ive high speed turning process. Higher cuttingparameters would be able to increase the productivity and reduce the production costs. The effects of the cutting parameters and on the variables object of study were investigated and the role of the different lubri-cooling conditions was assessed.

Differences in clinical outcome between docetaxel and abiraterone acetate as the first-line treatment in chemo-naïve metastatic castration-resistant prostate cancer patients with or without the ineligible clinical factors of the COU-AA-302 study.

PubMed

Poon, Darren M C; Chan, Kuen; Lee, Siu H; Chan, Tim W; Sze, Henry; Lee, Eric K C; Lam, Daisy; Chan, Michelle F T

2018-03-01

This study aimed to compare the efficacy of abiraterone acetate (AA) versus docetaxel (T) as first-line treatment in chemo-naïve metastatic castration-resistant prostate cancer (mCRPC) patients with or without the ineligible factors of the COU-AA-302 study (presence of visceral metastases, symptomatic disease, and/or Eastern Cooperative Oncology Group performance status ≥ 2). The clinical records of chemo-naïve mCRPC patients who received AA in six public oncology centers or T in two of these centers between 2003 and 2014 were reviewed. The survival time was compared among four subgroups of patients: those with ineligible factors administered AA (Group Ineligible-AA) or T (Group Ineligible-T), and those without ineligible factors and administered AA (Group Eligible-AA) or T (Group Eligible-T). During the study period, we identified 115 mCRPC patients who received AA or T, among whom 29, 36, 29, and 21 patients were classified as Groups Ineligible-AA, Ineligible-T, Eligible-AA, and Eligible-T, respectively. Both Group Ineligible-AA and Group Eligible-AA had significantly longer progression-free survival (PFS) and similar overall survival (OS) as Group Ineligible-T and Group Eligible-T (Ineligible, PFS: 6.3 vs. 5.9 months, P  = 0.0234, OS: 7.8 vs. 15.7 months, P  = 0.1601; Eligible, PFS: 9.8 vs. 5.6 months, P  = 0.0437, OS: 20.5 vs. 18.2 months, P  = 0.7820). Compared to T, AA treatment resulted in longer PFS and similar OS in chemo-naïve mCRPC patients, irrespective of the presence of ineligible factors, suggesting that the initial treatment by AA may still be beneficial to those with the aforementioned ineligible factors.

TCR Signal Strength Regulates Akt Substrate Specificity To Induce Alternate Murine Th and T Regulatory Cell Differentiation Programs.

PubMed

Hawse, William F; Boggess, William C; Morel, Penelope A

2017-07-15

The Akt/mTOR pathway is a key driver of murine CD4 + T cell differentiation, and induction of regulatory T (Treg) cells results from low TCR signal strength and low Akt/mTOR signaling. However, strong TCR signals induce high Akt activity that promotes Th cell induction. Yet, it is unclear how Akt controls alternate T cell fate decisions. We find that the strength of the TCR signal results in differential Akt enzymatic activity. Surprisingly, the Akt substrate networks associated with T cell fate decisions are qualitatively different. Proteomic profiling of Akt signaling networks during Treg versus Th induction demonstrates that Akt differentially regulates RNA processing and splicing factors to drive T cell differentiation. Interestingly, heterogeneous nuclear ribonucleoprotein (hnRNP) L or hnRNP A1 are Akt substrates during Treg induction and have known roles in regulating the stability and splicing of key mRNAs that code for proteins in the canonical TCR signaling pathway, including CD3ζ and CD45. Functionally, inhibition of Akt enzymatic activity results in the dysregulation of splicing during T cell differentiation, and knockdown of hnRNP L or hnRNP A1 results in the lower induction of Treg cells. Together, this work suggests that a switch in substrate specificity coupled to the phosphorylation status of Akt may lead to alternative cell fates and demonstrates that proteins involved with alternative splicing are important factors in T cell fate decisions. Copyright © 2017 by The American Association of Immunologists, Inc.

The trajectory and atmospheric impact of asteroid 2014 AA

NASA Astrophysics Data System (ADS)

Farnocchia, Davide; Chesley, Steven R.; Brown, Peter G.; Chodas, Paul W.

2016-08-01

Near-Earth asteroid 2014 AA entered the Earth's atmosphere on 2014 January 2, only 21 h after being discovered by the Catalina Sky Survey. In this paper we compute the trajectory of 2014 AA by combining the available optical astrometry, seven ground-based observations over 69 min, and the International Monitoring System detection of the atmospheric impact infrasonic airwaves in a least-squares orbit estimation filter. The combination of these two sources of observations results in a tremendous improvement in the orbit uncertainties. The impact time is 3:05 UT with a 1σ uncertainty of 6 min, while the impact location corresponds to a west longitude of 44.2° and a latitude of 13.1° with a 1σ uncertainty of 140 km. The minimum impact energy estimated from the infrasound data and the impact velocity result in an estimated minimum mass of 22.6 t. By propagating the trajectory of 2014 AA backwards we find that the only window for finding precovery observations is for the three days before its discovery.

Parallel loss of nuclear-encoded mitochondrial aminoacyl-tRNA synthetases and mtDNA-encoded tRNAs in Cnidaria.

PubMed

Haen, Karri M; Pett, Walker; Lavrov, Dennis V

2010-10-01

Unlike most animal mitochondrial (mt) genomes, which encode a set of 22 transfer RNAs (tRNAs) sufficient for mt protein synthesis, those of cnidarians have only retained one or two tRNA genes. Whether the missing cnidarian mt-tRNA genes relocated outside the main mt chromosome or were lost remains unclear. It is also unknown what impact the loss of tRNA genes had on other components of the mt translational machinery. Here, we explored the nuclear genome of the cnidarian Nematostella vectensis for the presence of mt-tRNA genes and their corresponding mt aminoacyl-tRNA synthetases (mt-aaRS). We detected no candidates for mt-tRNA genes and only two mt-aaRS orthologs. At the same time, we found that all but one cytosolic aaRS appear to be targeted to mitochondria. These results indicate that the loss of mt-tRNAs in Cnidaria is genuine and occurred in parallel with the loss of nuclear-encoded mt-aaRS. Our phylogenetic analyses of individual aaRS revealed that although the nearly total loss of mt-aaRS is rare, aaRS gene deletion and replacement have occurred throughout the evolution of Metazoa.

Characterization of the radical-scavenging reaction of 2-O-substituted ascorbic acid derivatives, AA-2G, AA-2P, and AA-2S: a kinetic and stoichiometric study.

PubMed

Takebayashi, Jun; Tai, Akihiro; Gohda, Eiichi; Yamamoto, Itaru

2006-04-01

The aim of this study was to characterize the antioxidant activity of three ascorbic acid (AA) derivatives O-substituted at the C-2 position of AA: ascorbic acid 2-glucoside (AA-2G), ascorbic acid 2-phosphate (AA-2P), and ascorbic acid 2-sulfate (AA-2S). The radical-scavenging activities of these AA derivatives and some common low molecular-weight antioxidants such as uric acid or glutathione against 1,1-diphenyl-picrylhydrazyl (DPPH) radical, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+), or galvinoxyl radical were kinetically and stoichiometrically evaluated under pH-controlled conditions. Those AA derivatives slowly and continuously reacted with DPPH radical and ABTS+, but not with galvinoxyl radical. They effectively reacted with DPPH radical under acidic conditions and with ABTS+ under neutral conditions. In contrast, AA immediately quenched all species of radicals tested at all pH values investigated. The reactivity of Trolox, a water-soluble vitamin E analogue, was comparable to that of AA in terms of kinetics and stoichiometrics. Uric acid and glutathione exhibited long-lasting radical-scavenging activity against these radicals under certain pH conditions. The radical-scavenging profiles of AA derivatives were closer to those of uric acid and glutathione rather than to that of AA. The number of radicals scavenged by one molecule of AA derivatives, uric acid, or glutathione was equal to or greater than that by AA or Trolox under the appropriate conditions. These data suggest the potential usage of AA derivatives as radical scavengers.

Susceptibility of Spodoptera frugiperda and S. exigua to Bacillus thuringiensis Vip3Aa insecticidal protein.

PubMed

Chakroun, Maissa; Bel, Yolanda; Caccia, Silvia; Abdelkefi-Mesrati, Lobna; Escriche, Baltasar; Ferré, Juan

2012-07-01

The Vip3Aa protein is an insecticidal protein secreted by Bacillus thuringiensis during the vegetative stage of growth. The activity of this protein has been tested after different steps/protocols of purification using Spodoptera frugiperda as a control insect. The results showed that the Vip3Aa protoxin was stable and retained full toxicity after being subjected to common biochemical steps used in protein purification. Bioassays with the protoxin in S. frugiperda and S. exigua showed pronounced differences in LC(50) values when mortality was measured at 7 vs. 10d. At 7d most live larvae were arrested in their development. LC(50) values of "functional mortality" (dead larvae plus larvae remaining in the first instar), measured at 7d, were similar or even lower than the LC(50) values of mortality at 10d. This strong growth inhibition was not observed when testing the trypsin-activated protein (62 kDa) in either species. S. exigua was less susceptible than S. frugiperda to the protoxin form, with LC(50) values around 10-fold higher. However, both species were equally susceptible to the trypsin-activated form. Processing of Vip3Aa protoxin to the activated form was faster with S. frugiperda midgut juice than with S. exigua midgut juice. The results strongly suggest that the differences in the rate of activation of the Vip3Aa protoxin between both species are the basis for the differences in susceptibility towards the protoxin form. Copyright © 2012 Elsevier Inc. All rights reserved.

Characteristic evaluation of process parameters of friction stir welding of aluminium 2024 hybrid composites

NASA Astrophysics Data System (ADS)

Sadashiva, M.; Shivanand, H. K.; Vidyasagar, H. N.

2018-04-01

The Current work is aimed to investigate the effect of process parameters in friction stir welding of Aluminium 2024 base alloy and Aluminium 2024 matrix alloy reinforced with E Glass and Silicon Carbide reinforcements. The process involved a set of synthesis techniques incorporating stir casting methodology resulting in fabrication of the composite material. This composite material that is synthesized is then machined to obtain a plate of dimensions 100 mm * 50 mm * 6 mm. The plate is then friction stir welded at different set of parameters viz. the spindle speed of 600 rpm, 900 rpm and 1200 rpm and feed rate of 40 mm/min, 80 mm/min and 120 mm/min for analyzing the process capability. The study of the given set of parameters is predominantly important to understand the physics of the process that may lead to better properties of the joint, which is very much important in perspective to its use in advanced engineering applications, especially in aerospace domain that uses Aluminium 2024 alloy for wing and fuselage structures under tension.

Cyanidin-3-rutinoside increases glucose uptake by activating the PI3K/Akt pathway in 3T3-L1 adipocytes.

PubMed

Choi, Kyung Ha; Lee, Hyun Ah; Park, Mi Hwa; Han, Ji-Sook

2017-09-01

In this study, the effect of cyanidin-3-rutinoside (C3R) on glucose uptake by 3T3-L1 adipocytes was studied. C3R significantly increased glucose uptake, which was associated with enhanced plasma membrane glucose transporter type 4 (PM-GLUT4) expression in 3T3-L1 adipocytes. The potentiating effect of C3R on glucose uptake and PM-GLUT4 expression was related to enhanced phosphorylation of insulin receptor substrate 1 (IRS-1) and Akt, as well as augmented activation of phosphatidylinositol-3-kinase (PI3K) in the insulin signaling pathway. C3R induced glucose uptake was inhibited only by the PI3K inhibitor, but not by an AMPK inhibitor in 3T3-L1 adipocytes. Therefore, C3R likely up-regulates glucose uptake and PM-GLUT4 expression in 3T3-L1 adipocytes by activating the PI3K/Akt pathways. Copyright © 2017 Elsevier B.V. All rights reserved.

Dynamics of the active site loops in catalyzing aminoacylation reaction in seryl and histidyl tRNA synthetases.

PubMed

Dutta, Saheb; Kundu, Soumya; Saha, Amrita; Nandi, Nilashis

2018-03-01

Aminoacylation reaction is the first step of protein biosynthesis. The catalytic reorganization at the active site of aminoacyl tRNA synthetases (aaRSs) is driven by the loop motions. There remain lacunae of understanding concerning the catalytic loop dynamics in aaRSs. We analyzed the functional loop dynamics in seryl tRNA synthetase from Methanopyrus kandleri ( mk SerRS) and histidyl tRNA synthetases from Thermus thermophilus ( tt HisRS), respectively, using molecular dynamics. Results confirm that the motif 2 loop and other active site loops are flexible spots within the catalytic domain. Catalytic residues of the loops form a network of interaction with the substrates to form a reactive state. The loops undergo transitions between closed state and open state and the relaxation of the constituent residues occurs in femtosecond to nanosecond time scale. Order parameters are higher for constituent catalytic residues which form a specific network of interaction with the substrates to form a reactive state compared to the Gly residues within the loop. The development of interaction is supported from mutation studies where the catalytic domain with mutated loop exhibits unfavorable binding energy with the substrates. During the open-close motion of the loops, the catalytic residues make relaxation by ultrafast librational motion as well as fast diffusive motion and subsequently relax rather slowly via slower diffusive motion. The Gly residues act as a hinge to facilitate the loop closing and opening by their faster relaxation behavior. The role of bound water is analyzed by comparing implicit solvent-based and explicit solvent-based simulations. Loops fail to form catalytically competent geometry in absence of water. The present result, for the first time reveals the nature of the active site loop dynamics in aaRS and their influence on catalysis.

Identification of AaCASPS7, an effector caspase in Aedes albopictus.

PubMed

Feng, Lingyan; Liu, Hao; Li, Xiaomei; Qiao, Jialu; Wang, Shengya; Guo, Deyin; Liu, Qingzhen

2016-11-15

Aedes albopictus mosquito is a vector of various arboviruses and is becoming a significant threat to public health due to its rapid global expansion. Several reports suggest that apoptosis could be a factor limiting arbovirus infection in mosquitoes. Thus, it is significant to identify apoptosis pathway and study the correlation between apoptosis and virus infection in mosquitoes. Apoptosis is a type of programmed cell death that plays a vital role in immunity, development, and tissue homeostasis. Caspases are a family of conserved proteases playing important roles in apoptosis. In this study, we identified Aedes albopictus AaCASPS7, a caspase shared high identity with dipteran insect drICE orthologs. Phylogenetic analysis showed the closest relative of AaCASPS7 was Aedes aegypti AeCASPS7. AaCASPS7 displayed several features that were typical of an effector caspase and showed significant activity to effector caspase substrates. Aacasps7 transcripts were expressed ubiquitously in developmental and adult stages in Aedes albopictus mosquitoes. Transient expression of AaCASPS7 induced caspase-dependent apoptosis in C6/36 cells. Taken together the above data, this study identified a novel caspase, AaCASPS7, which might function as an apoptotic caspase. Further study the function of AaCASPS7 would facilitate better understanding the apoptotic mechanism in Aedes albopictus mosquito. Copyright © 2016 Elsevier B.V. All rights reserved.

Analysis of Widespread Fatigue Damage in Aerospace Structures

DTIC Science & Technology

1999-02-01

Fatigue in 2024 - T351 Aluminum Alloy ," Wear, 221(1), pp 24-36 (1998). 20. T.N. Farris, M.P. Szolwinski and G...Fretting Fatigue in 2024 - T351 Aluminum Alloy ," Wear, 221(1), pp 24-36 (1998). Hsing-Ling Wang1, and Alten F. Grandt, Jr.2 FATIGUE ANALYSIS OF MULTIPLE...34 Effect of Prior Corrosion on the S/N Fatigue Performance of Aluminum Sheet Alloys 2024 -T3 and 2524-T3, Effects of the

Annonaceous acetogenin mimic AA005 induces cancer cell death via apoptosis inducing factor through a caspase-3-independent mechanism.

PubMed

Han, Bing; Wang, Tong-Dan; Shen, Shao-Ming; Yu, Yun; Mao, Chan; Yao, Zhu-Jun; Wang, Li-Shun

2015-03-18

Annonaceous acetogenins are a family of natural products with antitumor activities. Annonaceous acetogenin mimic AA005 reportedly inhibits mammalian mitochondrial NADH-ubiquinone reductase (Complex I) and induces gastric cancer cell death. However, the mechanisms underlying its cell-death-inducing activity are unclear. We used SW620 colorectal adenocarcinoma cells to study AA005 cytotoxic activity. Cell deaths were determined by Trypan blue assay and flow cytometry, and related proteins were characterized by western blot. Immunofluorescence and subcellular fractionation were used to evaluate AIF nuclear translocation. Reactive oxygen species were assessed by using redox-sensitive dye DCFDA. AA005 induces a unique type of cell death in colorectal adenocarcinoma cells, characterized by lack of caspase-3 activation or apoptotic body formation, sensitivity to poly (ADP-ribose) polymerase inhibitor Olaparib (AZD2281) but not pan-caspase inhibitor Z-VAD.fmk, and dependence on apoptosis-inducing factor (AIF). AA005 treatment also reduced expression of mitochondrial Complex I components, and leads to accumulation of intracellular reactive oxygen species (ROS) at the early stage. Blocking ROS formation significantly suppresses AA005-induced cell death in SW620 cells. Moreover, blocking activation of RIP-1 by necroptosis inhibitor necrotatin-1 inhibits AIF translocation and partially suppresses AA005-induced cell death in SW620 cells demonstrating that RIP-1 protein may be essential for cell death. AA005 may trigger the cell death via mediated by AIF through caspase-3 independent pathway. Our work provided new mechanisms for AA005-induced cancer cell death and novel clues for cancer treatment via AIF dependent cell death.

Homocysteine induced cardiovascular events: a consequence of long term anabolic‐androgenic steroid (AAS) abuse

PubMed Central

Graham, M R; Grace, F M; Boobier, W; Hullin, D; Kicman, A; Cowan, D; Davies, B; Baker, J S

2006-01-01

Objectives The long term effects (>20 years) of anabolic‐androgenic steroid (AAS) use on plasma concentrations of homocysteine (HCY), folate, testosterone, sex hormone binding globulin (SHBG), free androgen index, urea, creatinine, haematocrit (HCT), vitamin B12, and urinary testosterone/epitestosterone (T/E) ratio, were examined in a cohort of self‐prescribing bodybuilders. Methods Subjects (n = 40) were divided into four distinct groups: (1) AAS users still using AAS (SU; n = 10); (2) AAS users abstinent from AAS administration for 3 months (SA; n = 10); (3) non‐drug using bodybuilding controls (BC; n = 10); and (4) sedentary male controls (SC; n = 10). Results HCY levels were significantly higher in SU compared with BC and SC (p<0.01), and with SA (p<0.05). Fat free mass was significantly higher in both groups of AAS users (p<0.01). Daily energy intake (kJ) and daily protein intake (g/day) were significantly higher in SU and SA (p<0.05) compared with BC and SC, but were unlikely to be responsible for the observed HCY increases. HCT concentrations were significantly higher in the SU group (p<0.01). A significant linear inverse relationship was observed in the SU group between SHBG and HCY (r = −0.828, p<0.01), indicating a possible influence of the sex hormones in determining HCY levels. Conclusions With mounting evidence linking AAS to adverse effects on some clotting factors, the significantly higher levels of HCY and HCT observed in the SU group suggest long term AAS users have increased risk of future thromboembolic events. PMID:16488899

Growth and patterning of laser ablated superconducting YBa2Cu3O7 films on LaAlO3 substrates

NASA Technical Reports Server (NTRS)

Warner, J. D.; Bhasin, K. B.; Varaljay, N. C.; Bohman, D. Y.; Chorey, C. M.

1989-01-01

A high quality superconducting film on a substrate with a low dielectric constant is desired for passive microwave circuit applications. In addition, it is essential that the patterning process does not effect the superconducting properties of the thin films to achieve the highest circuit operating temperatures. YBa2Cu3O7 superconducting films were grown on lanthanum aluminate substrates using laser ablation with resulting maximum transition temperature (T sub c) of 90 K. The films were grown on a LaAlO3 which was at 775 C and in 170 mtorr of oxygen and slowly cooled to room temperature in 1 atm of oxygen. These films were then processed using photolithography and a negative photoresist with an etch solution of bromine and ethanol. Results are presented on the effect of the processing on T(sub c) of the film and the microwave properties of the patterned films.

Very large array and green bank telescope observations of Orion B (NGC 2024, W12): photodissociation region properties and magnetic field

DOE Office of Scientific and Technical Information (OSTI.GOV)

Roshi, D. Anish; Goss, W. M.; Jeyakumar, S., E-mail: aroshi@nrao.edu, E-mail: mgoss@nrao.edu, E-mail: sjk@astro.ugto.mx

We present images of C110α and H110α radio recombination line (RRL) emission at 4.8 GHz and images of H166α, C166α, and X166α RRL emission at 1.4 GHz, observed toward the star-forming region NGC 2024. The 1.4 GHz image with angular resolution ∼70'' is obtained using Very Large Array (VLA) data. The 4.8 GHz image with angular resolution ∼17'' is obtained by combining VLA and Green Bank Telescope data in order to add the short and zero spacing data in the uv plane. These images reveal that the spatial distributions of C110α line emission is confined to the southern rim ofmore » the H II region close to the ionization front whereas the C166α line emission is extended in the north-south direction across the H II region. The LSR velocity of the C110α line is 10.3 km s{sup –1} similar to that of lines observed from molecular material located at the far side of the H II region. This similarity suggests that the photodissociation region (PDR) responsible for C110α line emission is at the far side of the H II region. The LSR velocity of C166α is 8.8 km s{sup –1}. This velocity is comparable with the velocity of molecular absorption lines observed from the foreground gas, suggesting that the PDR is at the near side of the H II region. Non-LTE models for carbon line-forming regions are presented. Typical properties of the foreground PDR are T {sub PDR} ∼ 100 K, n{sub e}{sup PDR}∼5 cm{sup –3}, n {sub H} ∼ 1.7 × 10{sup 4} cm{sup –3}, and path length l ∼ 0.06 pc, and those of the far side PDR are T {sub PDR} ∼ 200 K, n{sub e}{sup PDR}∼ 50 cm{sup –3}, n {sub H} ∼ 1.7 × 10{sup 5} cm{sup –3}, and l ∼ 0.03 pc. Our modeling indicates that the far side PDR is located within the H II region. We estimate the magnetic field strength in the foreground PDR to be 60 μG and that in the far side PDR to be 220 μG. Our field estimates compare well with the values obtained from OH Zeeman observations toward NGC 2024. The H166α spectrum shows

Identification of Arabidopsis MYB56 as a novel substrate for CRL3(BPM) E3 ligases.

PubMed

Chen, Liyuan; Bernhardt, Anne; Lee, JooHyun; Hellmann, Hanjo

2015-02-01

Controlled stability of proteins is a highly efficient mechanism to direct diverse processes in living cells. A key regulatory system for protein stability is given by the ubiquitin proteasome pathway, which uses E3 ligases to mark specific proteins for degradation. In this work, MYB56 is identified as a novel target of a CULLIN3 (CUL3)-based E3 ligase. Its stability depends on the presence of MATH-BTB/POZ (BPM) proteins, which function as substrate adaptors to the E3 ligase. Genetic studies have indicated that MYB56 is a negative regulator of flowering, while BPMs positively affect this developmental program. The interaction between BPMs and MYB56 occurs at the promoter of FLOWERING LOCUS T (FT), a key regulator in initiating flowering in Arabidopsis, and results in instability of MYB56. Overall the work establishes MYB transcription factors as substrates of BPM proteins, and provides novel information on components that participate in controlling flowering time in plants. Copyright © 2015 The Author. Published by Elsevier Inc. All rights reserved.

Nonenzymatic template-directed synthesis on hairpin oligonucleotides. 3. Incorporation of adenosine and uridine residues

NASA Technical Reports Server (NTRS)

Wu, T.; Orgel, L. E.

1992-01-01

We have used [32P]-labeled hairpin oligonucleotides to study template-directed synthesis on templates containing one or more A or T residues within a run of C residues. When nucleoside-5'-phosphoro(2-methyl)imidazolides are used as substrates, isolated A and T residues function efficiently in facilitating the incorporation of U and A, respectively. The reactions are regiospecific, producing mainly 3'-5'-phosphodiester bonds. Pairs of consecutive non-C residues are copied much less efficiently. Limited synthesis of CA and AC sequences on templates containing TG and GT sequences was observed along with some synthesis of the AA sequences on templates containing TT sequences. The other dimer sequences investigated, AA, AG, GA, TA, and AT, could not be copied. If A is absent from the reaction mixture, misincorporation of G residues is a significant reaction on templates containing an isolated T residue or two consecutive T residues. However, if both A and G are present, A is incorporated to a much greater extent than G. We believe that wobble-pairing between T and G is responsible for misincorporation when only G is present.

Phosphoinositide 3–kinase γ participates in T cell receptor–induced T cell activation

PubMed Central

Alcázar, Isabela; Marqués, Miriam; Kumar, Amit; Hirsch, Emilio; Wymann, Matthias; Carrera, Ana C.; Barber, Domingo F.

2007-01-01

Class I phosphoinositide 3–kinases (PI3Ks) constitute a family of enzymes that generates 3-phosphorylated polyphosphoinositides at the cell membrane after stimulation of protein tyrosine (Tyr) kinase–associated receptors or G protein–coupled receptors (GPCRs). The class I PI3Ks are divided into two types: class IA p85/p110 heterodimers, which are activated by Tyr kinases, and the class IB p110γ isoform, which is activated by GPCR. Although the T cell receptor (TCR) is a protein Tyr kinase–associated receptor, p110γ deletion affects TCR-induced T cell stimulation. We examined whether the TCR activates p110γ, as well as the consequences of interfering with p110γ expression or function for T cell activation. We found that after TCR ligation, p110γ interacts with Gαq/11, lymphocyte-specific Tyr kinase, and ζ-associated protein. TCR stimulation activates p110γ, which affects 3-phosphorylated polyphosphoinositide levels at the immunological synapse. We show that TCR-stimulated p110γ controls RAS-related C3 botulinum substrate 1 activity, F-actin polarization, and the interaction between T cells and antigen-presenting cells, illustrating a crucial role for p110γ in TCR-induced T cell activation. PMID:17998387

Somatic Hypermutation at A/T-Rich Oligonucleotide Substrates Shows Different Strand Polarities in Ung-Deficient or -Proficient Backgrounds

PubMed Central

Zivojnovic, Marija; Delbos, Frédéric; Girelli Zubani, Giulia; Julé, Amélie; Alcais, Alexandre; Storck, Sébastien

2014-01-01

A/T mutations at immunoglobulin loci are introduced by DNA polymerase η (Polη) during an Msh2/6-dependent repair process which results in A's being mutated 2-fold more often than T's. This patch synthesis is initiated by a DNA incision event whose origin is still obscure. We report here the analysis of A/T oligonucleotide mutation substrates inserted at the heavy chain locus, including or not including internal C's or G's. Surprisingly, the template composed of only A's and T's was highly mutated over its entire 90-bp length, with a 2-fold decrease in mutation from the 5′ to the 3′ end and a constant A/T ratio of 4. These results imply that Polη synthesis was initiated from a break in the 5′-flanking region of the substrate and proceeded over its entire length. The A/T bias was strikingly altered in an Ung−/− background, which provides the first experimental evidence supporting a concerted action of Ung and Msh2/6 pathways to generate mutations at A/T bases. New analysis of Pms2−/− animals provided a complementary picture, revealing an A/T mutation ratio of 4. We therefore propose that Ung and Pms2 may exert a mutual backup function for the DNA incision that promotes synthesis by Polη, each with a distinct strand bias. PMID:24710273

Antibacterial and cytotoxic activity of isoprenylated coumarin mammea A/AA isolated from Mammea africana.

PubMed

Canning, Corene; Sun, Shi; Ji, Xiangming; Gupta, Smiti; Zhou, Kequan

2013-05-02

The stem bark of Mammea africana is widely distributed in tropical Africa and commonly used in traditional medicine. This study aims to identify the active compound in Mammea africana and to evaluate its antimicrobial and antiproliferative activity. Methanol extract from the bark of the Mammea africana was separated by liquid-liquid extraction, followed by open column chromatography. A principal antimicrobial compound was purified by high performance liquid chromatography (HPLC) and its structure was elucidated by nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS). The antibacterial activity of the purified compound was determined using the broth microdilution method against 7 common pathogenic bacteria. The compound was also evaluated for cytotoxicity by cell proliferation assay (MTS) using the mouse embryonic fibroblast cell line NIH 3T3 and the non-small cell lung cancer cell line A549. The purified active compound was determined to be mammea A/AA and was found to be highly active against Campylobacter jejuni (MIC=0.5 μg/ml), Streptococcus pneumoniae (MIC=0.25 μg/ml), and Clostridium difficile (MIC=0.25 μg/ml). The compound exhibited significant antiproliferative activities against both NIH 3T3 and A549 cell lines. Mammea A/AA isolated from Mammea africana exerts specific inhibitory activity against Campylobacter jejuni, Streptococcus pneumoniae, and Campylobacter difficile. Mammea A/AA was also found to exhibit significant cytotoxicity against both cancer and normal cell lines. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Residual stress analysis on AA6061+22% Al2O3p simple shape demonstrators of a wheel hub

NASA Astrophysics Data System (ADS)

Giuliani, Alessandra; Albertini, Gianni; Fiori, Fabrizio; Girardin, Emmanuelle

2004-07-01

The effects of thermal treatments in a component for automotive industry are considered: the induced residual stresses in wheel hubs were investigated. Measurements have been carried out on two identical simple shape demonstrators made of AA6061+22% Al2O3p in order to evaluate the stress conditions after forging and T6 treatment and without taking into account the final design of the wheel hub. One sample was analysed in as-forged conditions while the second after forging and T6 treatment (560°C for 2h, 20°C water quenching, artificial aging at 177°C for 10h). Particular attention was given to the stresses in the radial and hoop directions, as the main consequences of the in-service fatigue are expected to be related to stresses along these two directions.

Structure of a novel antibacterial toxin that exploits elongation factor Tu to cleave specific transfer RNAs

DOE PAGES

Michalska, Karolina; Gucinski, Grant C.; Garza-Sanchez, Fernando; ...

2017-08-11

Contact-dependent growth inhibition (CDI) is a mechanism of inter-cellular competition in which Gram-negative bacteria exchange polymorphic toxins using type V secretion systems. Here, we present structures of the CDI toxin from Escherichia coli NC101 in ternary complex with its cognate immunity protein and elongation factor Tu (EF-Tu). The toxin binds exclusively to domain 2 of EF-Tu, partially overlapping the site that interacts with the 3'-end of aminoacyl-tRNA (aa-tRNA). The toxin exerts a unique ribonuclease activity that cleaves the single-stranded 3'-end from tRNAs that contain guanine discriminator nucleotides. EF-Tu is required to support this tRNase activity in vitro, suggesting the toxinmore » specifically cleaves substrate in the context of GTP·EF-Tu·aa-tRNA complexes. However, superimposition of the toxin domain onto previously solved GTP·EF-Tu·aa-tRNA structures reveals potential steric clashes with both aa-tRNA and the switch I region of EF-Tu. Further, the toxin induces conformational changes in EF-Tu, displacing a β-hairpin loop that forms a critical salt-bridge contact with the 3'-terminal adenylate of aa-tRNA. Altogether, these observations suggest that the toxin remodels GTP·EF-Tu·aa-tRNA complexes to free the 3'-end of aa-tRNA for entry into the nuclease active site.« less

Structure of a novel antibacterial toxin that exploits elongation factor Tu to cleave specific transfer RNAs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Michalska, Karolina; Gucinski, Grant C.; Garza-Sanchez, Fernando

Contact-dependent growth inhibition (CDI) is a mechanism of inter-cellular competition in which Gram-negative bacteria exchange polymorphic toxins using type V secretion systems. Here, we present structures of the CDI toxin from Escherichia coli NC101 in ternary complex with its cognate immunity protein and elongation factor Tu (EF-Tu). The toxin binds exclusively to domain 2 of EF-Tu, partially overlapping the site that interacts with the 3'-end of aminoacyl-tRNA (aa-tRNA). The toxin exerts a unique ribonuclease activity that cleaves the single-stranded 3'-end from tRNAs that contain guanine discriminator nucleotides. EF-Tu is required to support this tRNase activity in vitro, suggesting the toxinmore » specifically cleaves substrate in the context of GTP·EF-Tu·aa-tRNA complexes. However, superimposition of the toxin domain onto previously solved GTP·EF-Tu·aa-tRNA structures reveals potential steric clashes with both aa-tRNA and the switch I region of EF-Tu. Further, the toxin induces conformational changes in EF-Tu, displacing a β-hairpin loop that forms a critical salt-bridge contact with the 3'-terminal adenylate of aa-tRNA. Altogether, these observations suggest that the toxin remodels GTP·EF-Tu·aa-tRNA complexes to free the 3'-end of aa-tRNA for entry into the nuclease active site.« less

Natural Functions of PLIN2 Mediating Wnt/LiCl Signaling and Glycogen Synthase Kinase 3 (GSK3)/GSK3 Substrate-Related Effects Are Modulated by Lipid

PubMed Central

Lu, Xinyue; Song, Kaimei

2015-01-01

Belonging to the PLIN family, PLIN2 associates with lipid storage droplets (LSDs), but other functions of PLIN2 remain unclear. Here, we suggest that PLIN2 mediates Wnt signaling because PLIN2 small interfering RNA (siRNA) suppresses activation of Wnt/coreceptor pathways. The mediation in the Wnt/Frizzled pathway seems to occur from Dishevelleds to axin/glycogen synthase kinase 3(GSK3)/β-catenin complexes (AGβC) as Wnt decreases Dishevelled/PLIN2 but increases AGβC/PLIN2 associations. Augmenting cellular LSDs that affect PLIN2 associations with these proteins, oleic acid (OA) treatment inhibits Wnt-increased AGβC/PLIN2 associations and β-catenin T-cell factor signaling (β-CTS). Revealing that PLIN2 is a GSK3-associated protein, the study explored PLIN2-mediated effects on GSK3/GSK3 substrates. PLIN2 siRNA reduces inhibitory GSK3 levels and lithium chloride (LiCl)-upregulated β-catenin or CCAAT/enhancer binding protein α (c/EBPα) expression. OA treatment decreases LiCl-increased c/EBPα via PLIN2-c/EBPα dissociation. In addition to PLIN2 overexpression increasing β-CTS, PLIN2 depletion or overexpression drops or adds expression of GSK3 substrates, such as β-catenin, c/EBPα,c-Myc, cyclin D1, and insulin receptor substrate 1, and cell growth/survival. PLIN2 N or C terminus overexpression that is associated with higher levels of the substrates suggests that those substrates bind to specific regions of PLIN2. Mimicking the possible high lipid concentrations in cells in the human body under conditions of hyperlipidemia/obesity, OA-treated cells gain or reduce GSK3 substrate expression in parallel with a decrease (a Wnt-like effect) or increase in GSK3 activity, likely regulated by GSK3/PLIN2/GSK3 substrate associations. PMID:26598603

Corrosion behavior of aluminum-alumina composites in aerated 3.5 percent chloride solution

NASA Astrophysics Data System (ADS)

Acevedo Hurtado, Paul Omar

Aluminum based metal matrix composites are finding many applications in engineering. Of these Al-Al2O3 composites appear to have promise in a number of defense applications because of their mechanical properties. However, their corrosion behavior remains suspect, especially in marine environments. While efforts are being made to improve the corrosion resistance of Al-Al2O3 composites, the mechanism of corrosion is not well known. In this study, the corrosion behavior of powder metallurgy processed Al-Cu alloy reinforced with 10, 15, 20 and 25 vol. % Al2O3 particles (XT 1129, XT 2009, XT 2048, XT 2031) was evaluated in aerated 3.5% NaCl solution using microstructural and electrochemical measurements. AA1100-O and AA2024T4 monolithic alloys were also studied for comparison purposes. The composites and unreinforced alloys were subjected to potentiodynamic polarization and Electrochemical Impedance Spectroscopy (EIS) testing. Addition of 25 vol. % Al2O 3 to the base alloys was found to increase its corrosion resistance considerably. Microstructural studies revealed the presence of intermetallic Al2Cu particles in these composites that appeared to play an important role in the observations. Pitting potential for these composites was near corrosion potential values, and repassivation potential was below the corresponding corrosion potential, indicating that these materials begin to corrode spontaneously as soon as they come in contact with the 3.5 % NaCl solution. EIS measurements indicate the occurrence of adsorption/diffusion phenomena at the interface of the composites which ultimately initiate localized or pitting corrosion. Polarization resistance values were extracted from the EIS data for all the materials tested. Electrically equivalent circuits are proposed to describe and substantiate the corrosive processes occurring in these Al-Al2O 3 composite materials.

Investigation into the stress corrosion cracking properties of AA2099, an aluminum-lithium-copper alloy

NASA Astrophysics Data System (ADS)

Padgett, Barbara Nicole

Recently developed Al-Li-Cu alloys show great potential for implementation in the aerospace industry because of the attractive mix of good mechanical properties and low density. AA2099 is an Al-Li-Cu alloy with the following composition Al-2.69wt%Cu-1.8wt%Li-0.6wt%Zn-0.3wt%Mg-0.3wt%Mn-0.08wt%Zr. The environmental assisted cracking and localized corrosion behavior of the AA2099 was investigated in this thesis. The consequences of uncontrolled grain boundary precipitation via friction stir welding on the stress corrosion cracking (SCC) behavior of AA2099 was investigated first. Using constant extension rate testing, intergranular corrosion immersion experiments, and potentiodynamic scans, the heat-affected zone on the trailing edge of the weld (HTS) was determined to be most susceptible of the weld zones. The observed SCC behavior for the HTS was linked to the dissolution of an active phase (Al2CuLi, T1) populating the grain boundary. It should be stated that the SCC properties of AA2099 in the as-received condition were determined to be good. Focus was then given to the electrochemical behavior of precipitate phases that may occupy grain and sub-grain boundaries in AA2099. The grain boundary micro-chemistry and micro-electrochemistry have been alluded to within the literature as having significant influence on the SCC behavior of Al-Li-Cu alloys. Major precipitates found in this alloy system are T1 (Al 2CuLi), T2 (Al7.5Cu4Li), T B (Al6CuLi3), and theta (Al2 Cu). These phases were produced in bulk form so that the electrochemical nature of each phase could be characterized. It was determined T1 was most active electrochemically and theta was least. When present on grain boundaries in the alloy, electrochemical behavior of the individual precipitates aligned with the observed corrosion behavior of the alloy (e.g. TB was accompanied by general pitting corrosion and T 1 was accompanied by intergranular corrosion attack). In addition to the electrochemical behavior of

Symmetry and geometry considerations of atom transfer: deoxygenation of (silox)3WNO and R3PO (R = Me, Ph, (t)Bu) by (silox)3M (M = V, NbL (L = PMe3, 4-picoline), Ta; silox = (t)Bu3SiO).

PubMed

Veige, Adam S; Slaughter, LeGrande M; Lobkovsky, Emil B; Wolczanski, Peter T; Matsunaga, Nikita; Decker, Stephen A; Cundari, Thomas R

2003-10-06

Deoxygenations of (silox)(3)WNO (12) and R(3)PO (R = Me, Ph, (t)Bu) by M(silox)(3) (1-M; M = V, NbL (L = PMe(3), 4-picoline), Ta; silox = (t)Bu(3)SiO) reflect the consequences of electronic effects enforced by a limiting steric environment. 1-Ta rapidly deoxygenated R(3)PO (23 degrees C; R = Me (DeltaG degrees (rxn)(calcd) = -47 kcal/mol), Ph) but not (t)Bu(3)PO (85 degrees, >2 days), and cyclometalation competed with deoxygenation of 12 to (silox)(3)WN (11) and (silox)(3)TaO (3-Ta; DeltaG degrees (rxn)(calcd) = -100 kcal/mol). 1-V deoxygenated 12 slowly and formed stable adducts (silox)(3)V-OPR(3) (3-OPR(3)) with OPR(3). 1-Nb(4-picoline) (S = 0) and 1-NbPMe(3) (S = 1) deoxygenated R(3)PO (23 degrees C; R = Me (DeltaG degrees (rxn)(calcd from 1-Nb) = -47 kcal/mol), Ph) rapidly and 12 slowly (DeltaG degrees (rxn)(calcd) = -100 kcal/mol), and failed to deoxygenate (t)Bu(3)PO. Access to a triplet state is critical for substrate (EO) binding, and the S --> T barrier of approximately 17 kcal/mol (calcd) hinders deoxygenations by 1-Ta, while 1-V (S = 1) and 1-Nb (S --> T barrier approximately 2 kcal/mol) are competent. Once binding occurs, significant mixing with an (1)A(1) excited state derived from population of a sigma-orbital is needed to ensure a low-energy intersystem crossing of the (3)A(2) (reactant) and (1)A(1) (product) states. Correlation of a reactant sigma-orbital with a product sigma-orbital is required, and the greater the degree of bending in the (silox)(3)M-O-E angle, the more mixing energetically lowers the intersystem crossing point. The inability of substrates EO = 12 and (t)Bu(3)PO to attain a bent 90 degree angle M-O-E due to sterics explains their slow or negligible deoxygenations. Syntheses of relevant compounds and ramifications of the results are discussed. X-ray structural details are provided for 3-OPMe(3) (90 degree angle V-O-P = 157.61(9) degrees), 3-OP(t)Bu(3) ( 90 degree angle V-O-P = 180 degrees ), 1-NbPMe(3), and (silox)(3)ClWO (9).

Migration speed and directionality switch of normal epithelial cells after TGF-β1-induced EMT (tEMT) on micro-structured polydimethylsiloxane (PDMS) substrates with variations in stiffness and topographic patterning.

PubMed

Wu, Tsung-Hsien; Li, Chia-Hui; Tang, Ming-Jer; Liang, Jen-I; Chen, Chia-Hsin; Yeh, Ming-Long

2013-10-01

The epithelial to mesenchymal transition (EMT) involves several physiological and pathological phenomena and endows cells with invasive and migratory properties. However, the effects of substrate stiffness and topography on the migration of cells before or after transforming growth factor-β1 (TGF-β1)-induced EMT (tEMT) are unknown. Herein, we seed control or tEMT NMuMG cells on the 2D patterns consisted of 1 μm or 5 μm line-widths and groove or cone patterns on either 2 MPa (1.96 ± 0.48 MPa) or 4 MPa (3.70 ± 0.74 MPa) polydimethylsiloxane (PDMS) substrates. After tEMT, the increased expression of α-SMA with vinculin in focal adhesion (FA) sites led to an acceleration of tEMT cell motility. On the 2 MPa substrate, the most influenced substrate was the 1 μm, cone-patterned substrate, where the tEMT cells' motility decelerated by 0.13 μm/min (36% slower than the cells on groove pattern). However, on the 5 μm, groove-patterned substrate, where the tEMT cells demonstrated the most rapid motility relative to the control cells, with an increment of 0.18 μm/min (100%). Among the different physical cues from substrate, the cone pattern could impede the migration speed of tEMT cells. Furthermore, we recommend the groove-patterned with a 5 μm line-width substrate as a useful tool to differentiate control and tEMT cells by migration speed.

Bacterial Heat Shock Protein GroEL (Hsp64) Exerts Immunoregulatory Effects on T Cells by Utilizing Apoptosis

PubMed Central

Nalbant, Ayten; Kant, Melis

2016-01-01

Aggregatibacter actinomycetemcomitans (Aa) expresses a 64-kDa GroEL protein belonging to the heat shock family of proteins. This protein has been shown to influence human host cells, but the apoptotic capacity of the GroEL protein regarding T cells is not yet known. The purpose of this study was to investigate the ability of A. actinomycetemcomitans GroEL (AaGroEL) protein to induce human peripheral blood T-cell apoptosis. Endogenous, purified AaGroEL protein was used as an antigen. In AaGroEL-treated T cells, the data indicated that phosphatidylserine exposure, an early apoptotic event, was dose- and time-dependent. The AaGroEL-treated T cells were also positive for active caspase-3 in a dose-dependent manner. The rate of AaGroEL-induced apoptosis was suppressed by the addition of the general caspase inhibitor Z-VAD-FMK. Furthermore, cleaved caspase-8 bands (40/36 kDa and 23 kDa) were identified in cells responding to AaGroEL. DNA fragmentation was also detected in the AaGroEL-treated T cells. Overall, we demonstrated that the endogenous GroEL from A. actinomycetemcomitans has the capacity to induce T-cell apoptosis. PMID:27736933

40 CFR 174.501 - Bacillus thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a...

Code of Federal Regulations, 2010 CFR

2010-07-01

... in corn and cotton; exemption from the requirement of a tolerance. 174.501 Section 174.501 Protection... thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a tolerance. Residues of Bacillus thuringiensis Vip3Aa proteins in or on corn or cotton are exempt from the requirement of a...

40 CFR 174.501 - Bacillus thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a...

Code of Federal Regulations, 2011 CFR

2011-07-01

... in corn and cotton; exemption from the requirement of a tolerance. 174.501 Section 174.501 Protection... thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a tolerance. Residues of Bacillus thuringiensis Vip3Aa proteins in or on corn or cotton are exempt from the requirement of a...

40 CFR 174.501 - Bacillus thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a...

Code of Federal Regulations, 2014 CFR

2014-07-01

... in corn and cotton; exemption from the requirement of a tolerance. 174.501 Section 174.501 Protection... thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a tolerance. Residues of Bacillus thuringiensis Vip3Aa proteins in or on corn or cotton are exempt from the requirement of a...

40 CFR 174.501 - Bacillus thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a...

Code of Federal Regulations, 2012 CFR

2012-07-01

... in corn and cotton; exemption from the requirement of a tolerance. 174.501 Section 174.501 Protection... thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a tolerance. Residues of Bacillus thuringiensis Vip3Aa proteins in or on corn or cotton are exempt from the requirement of a...

40 CFR 174.501 - Bacillus thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a...

Code of Federal Regulations, 2013 CFR

2013-07-01

... in corn and cotton; exemption from the requirement of a tolerance. 174.501 Section 174.501 Protection... thuringiensis Vip3Aa protein in corn and cotton; exemption from the requirement of a tolerance. Residues of Bacillus thuringiensis Vip3Aa proteins in or on corn or cotton are exempt from the requirement of a...

Experimental transmission of AA amyloidosis by injecting the AA amyloid protein into interleukin-1 receptor antagonist knockout (IL-1raKO) mice.

PubMed

Watanabe, K; Uchida, K; Chambers, J K; Tei, M; Shoji, A; Ushio, N; Nakayama, H

2015-05-01

The incidence of AA amyloidosis is high in humans with rheumatoid arthritis and several animal species, including cats and cattle with prolonged inflammation. AA amyloidosis can be experimentally induced in mice using severe inflammatory stimuli and a coinjection of AA amyloid; however, difficulties have been associated with transmitting AA amyloidosis to a different animal species, and this has been attributed to the "species barrier." The interleukin-1 receptor antagonist knockout (IL-1raKO) mouse, a rodent model of human rheumatoid arthritis, has been used in the transmission of AA amyloid. When IL-1raKO and BALB/c mice were intraperitoneally injected with mouse AA amyloid together with a subcutaneous pretreatment of 2% AgNO3, all mice from both strains that were injected with crude or purified murine AA amyloid developed AA amyloidosis. However, the amyloid index, which was determined by the intensity of AA amyloid deposition, was significantly higher in IL-1raKO mice than in BALB/c mice. When IL-1raKO and BALB/c mice were injected with crude or purified bovine AA amyloid together with the pretreatment, 83% (5/6 cases) and 38% (3/8 cases) of IL-1raKO mice and 17% (1/6 cases) and 0% (0/6 cases) of BALB/c mice, respectively, developed AA amyloidosis. Similarly, when IL-1raKO and BALB/c mice were injected with crude or purified feline AA amyloid, 33% (2/6 cases) and 88% (7/8 cases) of IL-1raKO mice and 0% (0/6 cases) and 29% (2/6 cases) of BALB/c mice, respectively, developed AA amyloidosis. These results indicated that IL-1raKO mice are a useful animal model for investigating AA amyloidogenesis. © The Author(s) 2014.

Optimization of the Mechanical Properties and Residual Stresses in 2024 Aluminum Alloy Through Heat Treatment

NASA Astrophysics Data System (ADS)

Araghchi, M.; Mansouri, H.; Vafaei, R.; Guo, Y.

2018-05-01

Residual stresses induced during quenching of aluminum alloys cause dimensional instability and distortion. In this study, the effects of different concentrations of polyalkylene glycol (PAG) quenchants on residual stresses and mechanical properties of 2024 aluminum alloy were investigated. Surface residual stresses were measured by using hole-drilling strain-gauge method. Also, mechanical properties and microstructure of the heat-treated samples were analyzed using hardness measurements, tensile tests, and transmission electron microscopy. Results showed that quenching into a 15% polymeric solution and aging at 190 °C for 12 h cause 50% reduction in residual stress as compared with quenching in water at 20 °C and naturally aging. Moreover, tensile strength decreased by 104 MPa ( 20%) in compared with the T6 sample.

In Situ Formation of Decavanadate-Intercalated Layered Double Hydroxide Films on AA2024 and their Anti-Corrosive Properties when Combined with Hybrid Sol Gel Films

PubMed Central

Wu, Junsheng; Peng, Dongdong; He, Yuntao; Du, Xiaoqiong; Zhang, Zhan; Zhang, Bowei; Li, Xiaogang; Huang, Yizhong

2017-01-01

A layered double hydroxide (LDH) film was formed in situ on aluminum alloy 2024 through a urea hydrolysis method, and a decavanadate-intercalated LDH (LDH-V) film fabricated through the dip coating method. The microstructural and morphological characteristics were investigated by scanning electron microscopy (SEM). The corrosion-resistant performance was analyzed by electrochemical impedance spectroscopy (EIS), scanning electrochemical microscopy (SECM), and a salt-spray test (SST).The SEM results showed that a complete and defect-free surface was formed on the LDH-VS film. The anticorrosion results revealed that the LDH-VS film had better corrosion-resistant properties than the LDH-S film, especially long-term corrosion resistance. The mechanism of corrosion protection was proposed to consist of the self-healing effect of the decavanadate intercalation and the shielding effect of the sol-gel film. PMID:28772785

In Vivo and In Vitro Binding of Vip3Aa to Spodoptera frugiperda Midgut and Characterization of Binding Sites by 125I Radiolabeling

PubMed Central

Chakroun, Maissa

2014-01-01

Bacillus thuringiensis vegetative insecticidal proteins (Vip3A) have been recently introduced in important crops as a strategy to delay the emerging resistance to the existing Cry toxins. The mode of action of Vip3A proteins has been studied in Spodoptera frugiperda with the aim of characterizing their binding to the insect midgut. Immunofluorescence histological localization of Vip3Aa in the midgut of intoxicated larvae showed that Vip3Aa bound to the brush border membrane along the entire apical surface. The presence of fluorescence in the cytoplasm of epithelial cells seems to suggest internalization of Vip3Aa or a fragment of it. Successful radiolabeling and optimization of the binding protocol for the 125I-Vip3Aa to S. frugiperda brush border membrane vesicles (BBMV) allowed the determination of binding parameters of Vip3A proteins for the first time. Heterologous competition using Vip3Ad, Vip3Ae, and Vip3Af as competitor proteins showed that they share the same binding site with Vip3Aa. In contrast, when using Cry1Ab and Cry1Ac as competitors, no competitive binding was observed, which makes them appropriate candidates to be used in combination with Vip3A proteins in transgenic crops. PMID:25002420

Identification of Arabidopsis MYB56 as a novel substrate for CRL3BPM E3 ligases.

PubMed

Chen, Liyuan; Bernhardt, Anne; Lee, JooHyun; Hellmann, Hanjo

2014-10-24

Controlled stability of proteins is a highly efficient mechanism to direct diverse processes in living cells. A key regulatory system for protein stability is given by the ubiquitin proteasome pathway, which uses E3 ligases to mark specific proteins for degradation. In this work MYB56 is identified as a novel target of a CULLIN3 (CUL3)-based E3 ligase. Its stability depends on the presence of MATH-BTB/POZ (BPM) proteins, which function as substrate adaptors to the E3 ligase. Genetic studies pointed out that MYB56 is a negative regulator of flowering, while BPMs positively affect this developmental program. The interaction between BPMs and MYB56 occurs at the promoter of FLOWERING LOCUS T (FT), a key regulator in initiating flowering in Arabidopsis, and results in instability of MYB56. Overall the work establishes MYB transcription factors as substrates of BPM proteins, and provides novel information on components that participate in controlling the flowering time point in plants. © The Author 2014. Published by the Molecular Plant Shanghai Editorial Office in association with Oxford University Press on behalf of CSPB and IPPE, SIBS, CAS.

Somatic hypermutation at A/T-rich oligonucleotide substrates shows different strand polarities in Ung-deficient or -proficient backgrounds.

PubMed

Zivojnovic, Marija; Delbos, Frédéric; Girelli Zubani, Giulia; Julé, Amélie; Alcais, Alexandre; Weill, Jean-Claude; Reynaud, Claude-Agnès; Storck, Sébastien

2014-06-01

A/T mutations at immunoglobulin loci are introduced by DNA polymerase η (Polη) during an Msh2/6-dependent repair process which results in A's being mutated 2-fold more often than T's. This patch synthesis is initiated by a DNA incision event whose origin is still obscure. We report here the analysis of A/T oligonucleotide mutation substrates inserted at the heavy chain locus, including or not including internal C's or G's. Surprisingly, the template composed of only A's and T's was highly mutated over its entire 90-bp length, with a 2-fold decrease in mutation from the 5' to the 3' end and a constant A/T ratio of 4. These results imply that Polη synthesis was initiated from a break in the 5'-flanking region of the substrate and proceeded over its entire length. The A/T bias was strikingly altered in an Ung(-/-) background, which provides the first experimental evidence supporting a concerted action of Ung and Msh2/6 pathways to generate mutations at A/T bases. New analysis of Pms2(-/-) animals provided a complementary picture, revealing an A/T mutation ratio of 4. We therefore propose that Ung and Pms2 may exert a mutual backup function for the DNA incision that promotes synthesis by Polη, each with a distinct strand bias. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Pyrrolysyl-tRNA Synthetase, an Aminoacyl-tRNA Synthetase for Genetic Code Expansion

DOE PAGES

Crnkovic, Ana; Suzuki, Tateki; Soll, Dieter; ...

2016-06-14

Genetic code expansion (GCE) has become a central topic of synthetic biology. GCE relies on engineered aminoacyl-tRNA synthetases (aaRSs) and a cognate tRNA species to allow codon reassignment by co-translational insertion of non-canonical amino acids (ncAAs) into proteins. Introduction of such amino acids increases the chemical diversity of recombinant proteins endowing them with novel properties. Such proteins serve in sophisticated biochemical and biophysical studies both in vitro and in vivo, they may become unique biomaterials or therapeutic agents, and they afford metabolic dependence of genetically modified organisms for biocontainment purposes. In the Methanosarcinaceae the incorporation of the 22nd genetically encodedmore » amino acid, pyrrolysine (Pyl), is facilitated by pyrrolysyl-tRNA synthetase (PylRS) and the cognate UAG-recognizing tRNAPyl. This unique aaRS•tRNA pair functions as an orthogonal translation system (OTS) in most model organisms. The facile directed evolution of the large PylRS active site to accommodate many ncAAs, and the enzyme’s anticodon-blind specific recognition of the cognate tRNAPyl make this system highly amenable for GCE purposes. The remarkable polyspecificity of PylRS has been exploited to incorporate >100 different ncAAs into proteins. Here we review the Pyl-OT system and selected GCE applications to examine the properties of an effective OTS.« less

Laser irradiation effects on the surface, structural and mechanical properties of Al-Cu alloy 2024

NASA Astrophysics Data System (ADS)

Yousaf, Daniel; Bashir, Shazia; Akram, Mahreen; kalsoom, Umm-i.-; Ali, Nisar

2014-02-01

Laser irradiation effects on surface, structural and mechanical properties of Al-Cu-Mg alloy (Al-Cu alloy 2024) have been investigated. The specimens were irradiated for various fluences ranging from 3.8 to 5.5 J/cm2 using an Excimer (KrF) laser (248 nm, 18 ns, 30 Hz) under vacuum environment. The surface and structural modifications of the irradiated targets have been investigated by scanning electron microscope (SEM) and X-ray diffractometer (XRD), respectively. SEM analysis reveals the formation of micro-sized craters along the growth of periodic surface structures (ripples) at their peripheries. The size of the craters initially increases and then decreases by increasing the laser fluence. XRD analysis shows an anomalous trend in the peak intensity and crystallite size of the specimen irradiated for various fluences. A universal tensile testing machine and Vickers microhardness tester were employed in order to investigate the mechanical properties of the irradiated targets. The changes in yield strength, ultimate tensile strength and microhardness were found to be anomalous with increasing laser fluences. The changes in the surface and structural properties of Al-Cu alloy 2024 after laser irradiation have been associated with the changes in mechanical properties.

Investigation of Fatigue Crack-Growth Resistance of Aluminum Alloys under Spectrum Loading.

DTIC Science & Technology

1983-04-01

Effects on Fatigue Crack Progation in 2024 -T3 Aluminum Alloy ," Eng. Frac. Mech, Vol...results ("a" from 6 to 13 mm) can be made (Figure 20a): 1. The 2XXX alloys 2020-T651, 2324-T39, and 2024 - T351 had longer spectrum fatigue lives than the...strength and spectrum life exists at all three maximum peak stress levels for the 2024 alloy , with the lower yield strength T351 condition having

Selective Laser Treatment on Cold-Sprayed Titanium Coatings: Numerical Modeling and Experimental Analysis

NASA Astrophysics Data System (ADS)

Carlone, Pierpaolo; Astarita, Antonello; Rubino, Felice; Pasquino, Nicola; Aprea, Paolo

2016-12-01

In this paper, a selective laser post-deposition on pure grade II titanium coatings, cold-sprayed on AA2024-T3 sheets, was experimentally and numerically investigated. Morphological features, microstructure, and chemical composition of the treated zone were assessed by means of optical microscopy, scanning electron microscopy, and energy dispersive X-ray spectrometry. Microhardness measurements were also carried out to evaluate the mechanical properties of the coating. A numerical model of the laser treatment was implemented and solved to simulate the process and discuss the experimental outcomes. Obtained results highlighted the key role played by heat input and dimensional features on the effectiveness of the treatment.

A homogeneous method to measure aminoacyl-tRNA synthetase aminoacylation activity using scintillation proximity assay technology.

PubMed

Macarrón, R; Mensah, L; Cid, C; Carranza, C; Benson, N; Pope, A J; Díez, E

2000-09-10

A new method to measure the aminoacylation of tRNA based upon the use of the scintillation proximity assay (SPA) technology has been developed. The assay detects incorporation of radiolabeled amino acids into cognate tRNA, catalyzed by a specific aminoacyl-tRNA synthetase (aaRS). Under acidic conditions, uncoated yttrium silicate SPA beads were found to bind tRNA aggregates, while the radiolabeled amino acid substrate remains in solution, resulting in good signal discrimination of these two species in the absence of any separation steps. The usefulness of this approach was demonstrated by measurement of steady-state kinetic constants and inhibitor binding constants for a range of aaRS enzymes in comparison with data from standard, trichloroacetic acid-precipitation-based assays. In all cases, the data were quantitatively comparable. Although the radioisotopic counting efficiency of the SPA method was less than that of standard liquid scintillation counting, the statistical performance (i.e., signal to background, variability, stability) of the SPA assays was at least equivalent to the separation-based methods. The assay was also shown to work well in miniaturized 384-well microtiter plate formats, resulting in considerable reagent savings. In summary, a new method to characterize aaRS activity is described that is faster and more amenable to high-throughput screening than traditional methods. Copyright 2000 Academic Press.

Membrane topology and identification of key residues of EaDAcT, a plant MBOAT with unusual substrate specificity.

PubMed

Tran, Tam N T; Shelton, Jennifer; Brown, Susan; Durrett, Timothy P

2017-10-01

Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) catalyzes the transfer of an acetyl group from acetyl-CoA to the sn-3 position of diacylglycerol to form 3-acetyl-1,2-diacyl-sn-glycerol (acetyl-TAG). EaDAcT belongs to a small, plant-specific subfamily of the membrane bound O-acyltransferases (MBOAT) that acylate different lipid substrates. Sucrose gradient density centrifugation revealed that EaDAcT colocalizes to the same fractions as an endoplasmic reticulum (ER)-specific marker. By mapping the membrane topology of EaDAcT, we obtained an experimentally determined topology model for a plant MBOAT. The EaDAcT model contains four transmembrane domains (TMDs), with both the N- and C-termini orientated toward the lumen of the ER. In addition, there is a large cytoplasmic loop between the first and second TMDs, with the MBOAT signature region of the protein embedded in the third TMD close to the interface between the membrane and the cytoplasm. During topology mapping, we discovered two cysteine residues (C187 and C293) located on opposite sides of the membrane that are important for enzyme activity. In order to identify additional amino acid residues important for acetyltransferase activity, we isolated and characterized acetyltransferases from other acetyl-TAG-producing plants. Among them, the acetyltransferase from Euonymus fortunei possessed the highest activity in vivo and in vitro. Mutagenesis of conserved amino acids revealed that S253, H257, D258 and V263 are essential for EaDAcT activity. Alteration of residues unique to the acetyltransferases did not alter the unique acyl donor specificity of EaDAcT, suggesting that multiple amino acids are important for substrate recognition. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

Resistance Risk Assessment of Spodoptera frugiperda (Lepidoptera: Noctuidae) and Diatraea saccharalis (Lepidoptera: Crambidae) to Vip3Aa20 Insecticidal Protein Expressed in Corn.

PubMed

Bernardi, Oderlei; Bernardi, Daniel; Amado, Douglas; Sousa, Renan S; Fatoretto, Julio; Medeiros, Fernanda C L; Conville, Jared; Burd, Tony; Omoto, Celso

2015-12-01

Transgenic Agrisure Viptera 3 corn that expresses Cry1Ab, Vip3Aa20, and EPSPS proteins and Agrisure Viptera expressing Vip3Aa20 are used for control of Spodoptera frugiperda (J.E. Smith) and Diatraea saccharalis (F.) in Brazil. To support a resistance management program, resistance risk assessment studies were conducted to characterize the dose expression of Vip3Aa20 protein and level of control against these species. The Vip3Aa20 expression in Agrisure Viptera 3 and Agrisure Viptera decreased from V6 to V10 stage of growth. However, Vip3Aa20 expression in Agrisure Viptera 3 at V6 and V10 stages was 13- and 16-fold greater than Cry1Ab, respectively. The Vip3Aa20 expression in lyophilized tissue of Agrisure Viptera 3 and Agrisure Viptera diluted 25-fold in an artificial diet caused complete larval mortality of S. frugiperda and D. saccharalis. In contrast, lyophilized tissue of Bt11 at the same dilution does not provide complete mortality of these species. Agrisure Viptera 3 and Agrisure Viptera also caused a high level of mortality against S. frugiperda and D. saccharalis. Moreover, 100% mortality was observed for S. frugiperda larvae (neonates through fifth-instar larvae) when fed in corn with the Vip trait technology. Viptera corn achieves a high level of control against S. frugiperda and D. saccharalis providing a high dose, which is an important determination to support the refuge strategy for an effective resistance management program. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Magnetoresistivity of thin YBa2Cu3O7-δ films on sapphire substrate

NASA Astrophysics Data System (ADS)

Probst, Petra; Il'in, Konstantin; Engel, Andreas; Semenov, Alexei; Hübers, Heinz-Wilhelm; Hänisch, Jens; Holzapfel, Bernhardt; Siegel, Michael

2012-09-01

Magnetoresistivity of YBa2Cu3O7-δ films with thicknesses between 7 and 100 nm deposited on CeO2 and PrBa2Cu3O7-δ buffer layers on sapphire substrate has been measured to analyze the temperature dependence of the second critical magnetic field Bc2. To define Bc2, the mean-field transition temperature Tc was evaluated by fitting the resistive transition in zero magnetic field with the fluctuation conductivity theory of Aslamazov and Larkin. At T → Tc the Bc2(T) dependence shows a crossover from downturn to upturn curvature with the increase in film thickness.

Aa Ah Nak

ERIC Educational Resources Information Center

Tha, Na Gya; Wus, Thay

2017-01-01

In this article, Aa Ah Nak, the authors' methodology presents not only various reflections but also diverse contradictions about the Aa Nii language as well as language revitalization. This article explores language foundation and how the Aa Nii language revitalization is inextricably linked to the genocide and resulting historic trauma pervasive…

Bacillus thuringiensis Cry1Ia10 and Vip3Aa protein interactions and their toxicity in Spodoptera spp. (Lepidoptera).

PubMed

Bergamasco, V B; Mendes, D R P; Fernandes, O A; Desidério, J A; Lemos, M V F

2013-02-01

The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65 kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana. Copyright © 2012 Elsevier Inc. All rights reserved.

Pitting corrosion inhibition of aluminum 2024 by Bacillus biofilms secreting polyaspartate or gamma-polyglutamate.

PubMed

Ornek, D; Jayaraman, A; Syrett, B C; Hsu, C-H; Mansfeld, F B; Wood, T K

2002-04-01

Pitting corrosion of aluminum 2024 in Luria Bertani medium was reduced by the secretion of anionic peptides by engineered and natural Bacillus biofilms and was studied in continuous reactors using electrochemical impedance spectroscopy. Compared to sterile controls, pitting was reduced dramatically by the presence of the biofilms. The secretion of a 20 amino acid polyaspartate peptide by an engineered Bacillus subtilis WB600/pBE92-Asp biofilm slightly reduced the corrosion rate of the passive aluminum alloy at pH 6.5; however, the secretion of gamma-polyglutamate by a Bacillus licheniformis biofilm reduced the corrosion rate by 90% (compared to the B. subtilis WB600/pBE92 biofilm which did not secrete polyaspartate or gamma-polyglutamate). The corrosion potential ( E(corr)) of aluminum 2024 was increased by about 0.15-0.44 V due to the formation of B. subtilis and B. licheniformis biofilms as compared to sterile controls. The increase of E(corr) and the observed prevention of pitting indicate that the pitting potential ( E(pit)) had increased. This result and the further decrease of corrosion rates for the passive aluminum alloy suggest that the rate of the anodic metal dissolution reaction was reduced by an inhibitor produced by the biofilms. Purified gamma-polyglutamate also decreased the corrosion rates of aluminum 2024.

Launcher Roadmap for the CrVI Substitution of Surface Treatments. Screening of Trivalent-Chromium Conversion Solutions and First Promising Results for Repair Applications on Aluminium Alloys

NASA Astrophysics Data System (ADS)

Debout, Vincent; Pettier, Sophie

2014-06-01

Airbus Defence and Space, Space System is involved in a global roadmap for launchers in order to substitute hexavalent chromium (CrVI) and Cadmium in the current surface treatments on metallic structures.Within this framework, a screening of trivalent chromium (CrIII) conversion solutions for touch-up applications has been carried out since this step is crucial to perform local application or to repair minor damages on launcher structures but it leads to higher risks of exposure for the workers.Three commercial CrIII conversion solutions have been evaluated on high performance aluminum alloys such as AA2024 T3 and AA7175 T7351 that are often used as structural materials.This preliminary investigation highlights the effect of surface preparation, rinsing and conversion process on the final corrosion performance of conversion coatings (CCs). The results are also discussed in terms of visual aspect and adhesion with new Cr-free primers.Two operating sets of parameters are identified with promising results that represent the first steps towards the development of a new Cr-free touch-up process.

High-Speed Friction Stir Welding of AA7075-T6 Sheet: Microstructure, Mechanical Properties, Micro-texture, and Thermal History

NASA Astrophysics Data System (ADS)

Zhang, Jingyi; Upadhyay, Piyush; Hovanski, Yuri; Field, David P.

2018-01-01

Friction stir welding (FSW) is a cost-effective and high-quality joining process for aluminum alloys (especially heat-treatable alloys) that is historically operated at lower joining speeds (up to hundreds of millimeters per minute). In this study, we present a microstructural analysis of friction stir welded AA7075-T6 blanks with high welding speeds up to 3 M/min. Textures, microstructures, mechanical properties, and weld quality are analyzed using TEM, EBSD, metallographic imaging, and Vickers hardness. The higher welding speed results in narrower, stronger heat-affected zones (HAZs) and also higher hardness in the nugget zones. The material flow direction in the nugget zone is found to be leaning towards the welding direction as the welding speed increases. Results are coupled with welding parameters and thermal history to aid in the understanding of the complex material flow and texture gradients within the welds in an effort to optimize welding parameters for high-speed processing.

Strengthening Mechanisms in Microtruss Metals

NASA Astrophysics Data System (ADS)

Ng, Evelyn K.

Microtrusses are hybrid materials composed of a three-dimensional array of struts capable of efficiently transmitting an externally applied load. The strut connectivity of microtrusses enables them to behave in a stretch-dominated fashion, allowing higher specific strength and stiffness values to be reached than conventional metal foams. While much attention has been given to the optimization of microtruss architectures, little attention has been given to the strengthening mechanisms inside the materials that make up this architecture. This thesis examines strengthening mechanisms in aluminum alloy and copper alloy microtruss systems with and without a reinforcing structural coating. C11000 microtrusses were stretch-bend fabricated for the first time; varying internal truss angles were selected in order to study the accumulating effects of plastic deformation and it was found that the mechanical performance was significantly enhanced in the presence of work hardening with the peak strength increasing by a factor of three. The C11000 microtrusses could also be significantly reinforced with sleeves of electrodeposited nanocrystalline Ni-53wt%Fe. It was found that the strength increase from work hardening and electrodeposition were additive over the range of structures considered. The AA2024 system allowed the contribution of work hardening, precipitation hardening, and hard anodizing to be considered as interacting strengthening mechanisms. Because of the lower formability of AA2024 compared to C11000, several different perforation geometries in the starting sheet were considered in order to more effectively distribute the plastic strain during stretch-bend fabrication. A T8 condition was selected over a T6 condition because it was shown that the plastic deformation induced during the final step was sufficient to enhance precipitation kinetics allowing higher strengths to be reached, while at the same time eliminating one annealing treatment. When hard anodizing

UCP2 and UCP3 variants and gene-environment interaction associated with prediabetes and T2DM in a rural population: a case control study in China.

PubMed

Su, Meifang; Chen, Xiaoying; Chen, Yue; Wang, Congyun; Li, Songtao; Ying, Xuhua; Xiao, Tian; Wang, Na; Jiang, Qingwu; Fu, Chaowei

2018-03-12

There are disparities for the association between uncoupling proteins (UCP) and type 2 diabetes (T2DM). The study was to examine the associations of genetic variants of UCP2 and UCP3 with prediabetes and T2DM in a rural Chinese population. A population-based case-control study of 397 adults with T2DM, 394 with prediabetes and 409 with normal glucose tolerance (NGT) was carried out in 2014 in a rural community in eastern China. Three groups were identified through a community survey and the prediabetes and NGT groups were frequently matched by age and gender with the T2DM group and they were not relatives of T2DM subjects. With r 2  ≥ 0.8 and minor allele frequency (MAF) ≥0.05 for tag single nucleotide polymorphisms (SNPs) with potential function, three (rs660339, rs45560234 and rs643064) and six (rs7930460, rs15763, rs647126, rs1800849, rs3781907 and rs1685356) SNPs were selected respectively for UCP2 and UCP3 and genotyped in real time using the MassARRAY system (Sequenom; USA). The haplotypes, gene-environmental interaction and association between genetic variants of UCP2 and UCP3 and prediabetes or T2DM were explored. There were no significant differences in age and sex among three study groups. After the adjustment for possible covariates, the A allele of rs1800849 in UCP3 was significantly associated with prediabetes (aOR AA vs GG  = 1.68, 95% CI: 1.02-2.78), and the association was also significant under the recessive model (aOR AA vs GA + GG  = 1.64, 95% CI: 1.02-2.66). Also, rs15763 was found to be marginally significantly associated with T2DM under dominant model (OR GA + AA vs GG  = 0.73, 95% CI: 0.52-1.03, P = 0.072). No haplotype was significantly associated with prediabetes or T2DM. Multiplicative interactions for rs660339-overweight on T2DM were observed. In addition, the AA genotype of rs660339 was associated with an increased risk of T2DM in overweight subjects (OR = 1.48, 95%CI: 0.87-2.52) but with a decreased

Abnormal Interactions between Perifollicular Mast Cells and CD8+ T-Cells May Contribute to the Pathogenesis of Alopecia Areata

PubMed Central

Bertolini, Marta; Zilio, Federica; Rossi, Alfredo; Gilhar, Amos; Keren, Aviad; Meyer, Katja C.; Wang, Eddy; Funk, Wolfgang; McElwee, Kevin; Paus, Ralf

2014-01-01

Alopecia areata (AA) is a CD8+ T-cell dependent autoimmune disease of the hair follicle (HF) in which the collapse of HF immune privilege (IP) plays a key role. Mast cells (MCs) are crucial immunomodulatory cells implicated in the regulation of T cell-dependent immunity, IP, and hair growth. Therefore, we explored the role of MCs in AA pathogenesis, focusing on MC interactions with CD8+ T-cells in vivo, in both human and mouse skin with AA lesions. Quantitative (immuno-)histomorphometry revealed that the number, degranulation and proliferation of perifollicular MCs are significantly increased in human AA lesions compared to healthy or non-lesional control skin, most prominently in subacute AA. In AA patients, perifollicular MCs showed decreased TGFβ1 and IL-10 but increased tryptase immunoreactivity, suggesting that MCs switch from an immuno-inhibitory to a pro-inflammatory phenotype. This concept was supported by a decreased number of IL-10+ and PD-L1+ MCs, while OX40L+, CD30L+, 4–1BBL+ or ICAM-1+ MCs were increased in AA. Lesional AA-HFs also displayed significantly more peri- and intrafollicular- CD8+ T-cells as well as more physical MC/CD8+ T-cell contacts than healthy or non-lesional human control skin. During the interaction with CD8+ T-cells, AA MCs prominently expressed MHC class I and OX40L, and sometimes 4–1BBL or ICAM-1, suggesting that MC may present autoantigens to CD8+ T-cells and/or co-stimulatory signals. Abnormal MC numbers, activities, and interactions with CD8+ T-cells were also seen in the grafted C3H/HeJ mouse model of AA and in a new humanized mouse model for AA. These phenomenological in vivo data suggest the novel AA pathobiology concept that perifollicular MCs are skewed towards pro-inflammatory activities that facilitate cross-talk with CD8+ T-cells in this disease, thus contributing to triggering HF-IP collapse in AA. If confirmed, MCs and their CD8+ T-cell interactions could become a promising new therapeutic target in the future

Effect of Al2O3sf addition on the friction and wear properties of (SiCp+Al2O3sf)/Al2024 composites fabricated by pressure infiltration

NASA Astrophysics Data System (ADS)

Xu, Hui; Zhang, Gong-zhen; Cui, Wei; Ren, Shu-bin; Wang, Qian-jin; Qu, Xuan-hui

2018-03-01

Aluminum (Al) 2024 matrix composites reinforced with alumina short fibers (Al2O3sf) and silicon carbide particles (SiCp) as wear-resistant materials were prepared by pressure infiltration in this study. Further, the effect of Al2O3sf on the friction and wear properties of the as-synthesized composites was systematically investigated, and the relationship between volume fraction and wear mechanism was discussed. The results showed that the addition of Al2O3sf, characterized by the ratio of Al2O3sf to SiCp, significantly affected the properties of the composites and resulted in changes in wear mechanisms. When the volume ratio of Al2O3sf to SiCp was increased from 0 to 1, the rate of wear mass loss ( K m) and coefficients of friction (COFs) of the composites decreased, and the wear mechanisms were abrasive wear and furrow wear. When the volume ratio was increased from 1 to 3, the COF decreased continuously; however, the K m increased rapidly and the wear mechanism became adhesive wear.

Mycoplasma-induced BALB/c 3T3 collagenase is a mammalian enzyme.

PubMed Central

Kluve, B; Merrick, W C; Gershman, H

1983-01-01

A collagenase previously reported to accumulate in the medium of cultures of BALB/c 3T3 cells on infection with Mycoplasma orale [Kluve, Merrick, Stanbridge & Gershman (1981) Nature (London) 292, 855-857] was partially purified and characterized. With regard to purification properties, activation, sensitivity to inhibitors and relative molecular mass the enzyme was similar to previously reported vertebrate collagenases, but could not be unequivocally distinguished from bacterial collagenases. With regard to substrate-specificity and reaction products, however, the collagenase was typical of vertebrate collagenases and distinct from bacterial collagenases. Specifically, the enzyme displayed a preference for type III collagen and type I collagen, a somewhat decreased ability to degrade type II collagen, and a very limited ability to degrade type IV collagen. The initial products of the action of the collagenase on type I collagen were characterized as fragments one-quarter and three-quarters of the length of the intact collagen molecule. Because the properties of the collagenase produced by cultures of mycoplasma-infected BALB/c 3T3 cells are those of a mammalian-type (vertebrate-type) enzyme, we have concluded that the collagenase is a product of the mouse (BALB/c 3T3) genome, and is not produced by the mycoplasma. Therefore it appears that infection of BALB/c 3T3 mouse fibroblasts with Mycoplasma orale induces the mouse cells to produce and secrete collagenase. PMID:6309150

A Multiple Aminoacyl-tRNA Synthetase Complex That Enhances tRNA-Aminoacylation in African Trypanosomes

PubMed Central

Cestari, Igor; Kalidas, Savitha; Monnerat, Severine; Anupama, Atashi; Phillips, Margaret A.

2013-01-01

The genes for all cytoplasmic and potentially all mitochondrial aminoacyl-tRNA synthetases (aaRSs) were identified, and all those tested by RNA interference were found to be essential for the growth of Trypanosoma brucei. Some of these enzymes were localized to the cytoplasm or mitochondrion, but most were dually localized to both cellular compartments. Cytoplasmic T. brucei aaRSs were organized in a multiprotein complex in both bloodstream and procyclic forms. The multiple aminoacyl-tRNA synthetase (MARS) complex contained at least six aaRS enzymes and three additional non-aaRS proteins. Steady-state kinetic studies showed that association in the MARS complex enhances tRNA-aminoacylation efficiency, which is in part dependent on a MARS complex-associated protein (MCP), named MCP2, that binds tRNAs and increases their aminoacylation by the complex. Conditional repression of MCP2 in T. brucei bloodstream forms resulted in reduced parasite growth and infectivity in mice. Thus, association in a MARS complex enhances tRNA-aminoacylation and contributes to parasite fitness. The MARS complex may be part of a cellular regulatory system and a target for drug development. PMID:24126051

Projections of Education Statistics to 2024. Forty-Third Edition. NCES 2016-013

ERIC Educational Resources Information Center

Hussar, William J.; Bailey, Tabitha M.

2016-01-01

"Projections of Education Statistics to 2024" is the 43rd report in a series begun in 1964. It includes statistics on elementary and secondary schools and degree-granting postsecondary institutions. This report provides revisions of projections shown in Projections of Education Statistics to 2023 and projections of enrollment, graduates,…

Long Non-Coding RNA SH3PXD2A-AS1 Promotes Cell Progression Partly Through Epigenetic Silencing P57 and KLF2 in Colorectal Cancer.

PubMed

Ma, Zhonghua; Peng, Peng; Zhou, Jing; Hui, Bingqing; Ji, Hao; Wang, Juan; Wang, Keming

2018-05-03

Colorectal cancer (CRC) is one of the most commonly diagnosed malignancies worldwide. Current evidence has revealed the key roles of long non-coding RNAs (IncRNAs) in multiple cancers, including CRC. In this study we identified the lncRNA SH3PXD2A-AS1 as a novel molecule associated with CRC progression by analyzing the publicly available data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. Quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed to examine the expression levels of SH3PXD2A-AS1 in CRC tissue samples and CRC cell lines. Cell viability examination, colony-formation experiments, ethynyl deoxyuridine (Edu) assays and flow cytometry were performed to investigate the roles of SH3PXD2A-AS1 in CRC proliferation, cell cycle regulation, and apoptosis. Transwell assays were used to explore the effects of SH3PXD2A-AS1 on CRC cells migration and invasion. A nude mice model was used to assess the effects of SH3PXD2A-AS1 on tumorigenesis in vivo. Subcellular fractionation, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP) assays were conducted to detect the molecular mechanisms of SH3PXD2A-ASl-mediated gene expression. Rescue assays were used to determine whether P57 and Kruppel-like factor 2 (KLF2) were involved in SH3PXD2A-ASl-dependent CRC proliferation. We firstly found that SH3PXD2A-AS1 was significantly upregulated in CRC tissues and cell lines, and overexpression of SH3PXD2A-AS1 was correlated with tumor size, TNM stage, and lymph node metastasis in patients with CRC. Furthermore, SH3PXD2A-AS1 knockdown inhibited CRC cells proliferation, migration and invasion in vitro, and suppressed tumorigenesis in vivo. Mechanistic studies indicated that SH3PXD2A-AS1 could epiqenetically repress P57 and KLF2 expression through interaction with EZH2. Rescue experiments suggested that SH3PXD2A-ASl-mediated oncogenesis was impaired by overexpression of P57 or KLF2. Interestingly, the expression

Fundamental Investigation of Fatigue Crack Growth Retardation in Aluminum Alloys

DTIC Science & Technology

1976-09-01

Fatigue Crack Propagation in 2024 -T3 Aluminum Alloy , " ASTM STP 536, p. 115, 1973. 9. J. Schijve, " Effect of Load Sequences...Hertzberg, " Effect of Multiple Over- loads on Fatigue Crack Propagation in 2024 -T3 Aluminum Alloy , " ASTM STP-536, p. 115, 1973. 9. J. Schijve... Effect of Thickness on Retardation Behavior of 7075 and 2024 Aluminum Alloys .......... 185 vi LIST OF ILLUSTRATIONS FIGURE PAGE 1 SEN

Substrate specificity of the aspartate:alanine antiporter (AspT) of Tetragenococcus halophilus in reconstituted liposomes.

PubMed

Sasahara, Ayako; Nanatani, Kei; Enomoto, Masaru; Kuwahara, Shigefumi; Abe, Keietsu

2011-08-19

The aspartate:alanine antiporter (AspT) of the lactic acid bacterium Tetragenococcus halophilus is a member of the aspartate:alanine exchanger (AAEx) transporter family. T. halophilus AspT catalyzes the electrogenic exchange of L-aspartate(1-) with L-alanine(0). Although physiological functions of AspT were well studied, L-aspartate(1-):L-alanine(0) antiport mechanisms are still unsolved. Here we report that the binding sites of L-aspartate and L-alanine are independently present in AspT by means of the kinetic studies. We purified His(6)-tagged T. halophilus AspT and characterized its kinetic properties when reconstituted in liposomes (K(m) = 0.35 ± 0.03 mm for L-aspartate, K(m) = 0.098 ± 0 mm for D-aspartate, K(m) = 26 ± 2 mm for L-alanine, K(m) = 3.3 ± 0.2 mm for D-alanine). Competitive inhibition by various amino acids of L-aspartate or L-alanine in self-exchange reactions revealed that L-cysteine selectively inhibited L-aspartate self-exchange but only weakly inhibited L-alanine self-exchange. Additionally, L-serine selectively inhibited L-alanine self-exchange but barely inhibited L-aspartate self-exchange. The aspartate analogs L-cysteine sulfinic acid, L-cysteic acid, and D-cysteic acid competitively and strongly inhibited L-aspartate self-exchange compared with L-alanine self-exchange. Taken together, these kinetic data suggest that the putative binding sites of L-aspartate and L-alanine are independently located in the substrate translocation pathway of AspT.

Unexpected expansion of tRNA substrate recognition by the yeast m1G9 methyltransferase Trm10.

PubMed

Swinehart, William E; Henderson, Jeremy C; Jackman, Jane E

2013-08-01

N-1 Methylation of the nearly invariant purine residue found at position 9 of tRNA is a nucleotide modification found in multiple tRNA species throughout Eukarya and Archaea. First discovered in Saccharomyces cerevisiae, the tRNA methyltransferase Trm10 is a highly conserved protein both necessary and sufficient to catalyze all known instances of m1G9 modification in yeast. Although there are 19 unique tRNA species that contain a G at position 9 in yeast, and whose fully modified sequence is known, only 9 of these tRNA species are modified with m1G9 in wild-type cells. The elements that allow Trm10 to distinguish between structurally similar tRNA species are not known, and sequences that are shared between all substrate or all nonsubstrate tRNAs have not been identified. Here, we demonstrate that the in vitro methylation activity of yeast Trm10 is not sufficient to explain the observed pattern of modification in vivo, as additional tRNA species are substrates for Trm10 m1G9 methyltransferase activity. Similarly, overexpression of Trm10 in yeast yields m1G9 containing tRNA species that are ordinarily unmodified in vivo. Thus, yeast Trm10 has a significantly broader tRNA substrate specificity than is suggested by the observed pattern of modification in wild-type yeast. These results may shed light onto the suggested involvement of Trm10 in other pathways in other organisms, particularly in higher eukaryotes that contain up to three different genes with sequence similarity to the single TRM10 gene in yeast, and where these other enzymes have been implicated in pathways beyond tRNA processing.

Troponin T3 regulates nuclear localization of the calcium channel Ca{sub v}β{sub 1a} subunit in skeletal muscle

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Tan; Taylor, Jackson; Jiang, Yang

The voltage-gated calcium channel (Ca{sub v}) β{sub 1a} subunit (Ca{sub v}β{sub 1a}) plays an important role in excitation–contraction coupling (ECC), a process in the myoplasm that leads to muscle-force generation. Recently, we discovered that the Ca{sub v}β{sub 1a} subunit travels to the nucleus of skeletal muscle cells where it helps to regulate gene transcription. To determine how it travels to the nucleus, we performed a yeast two-hybrid screening of the mouse fast skeletal muscle cDNA library and identified an interaction with troponin T3 (TnT3), which we subsequently confirmed by co-immunoprecipitation and co-localization assays in mouse skeletal muscle in vivo andmore » in cultured C2C12 muscle cells. Interacting domains were mapped to the leucine zipper domain in TnT3 COOH-terminus (160–244 aa) and Ca{sub v}β{sub 1a} NH{sub 2}-terminus (1–99 aa), respectively. The double fluorescence assay in C2C12 cells co-expressing TnT3/DsRed and Ca{sub v}β{sub 1a}/YFP shows that TnT3 facilitates Ca{sub v}β{sub 1a} nuclear recruitment, suggesting that the two proteins play a heretofore unknown role during early muscle differentiation in addition to their classical role in ECC regulation. - Highlights: • Previously, we demonstrated that Ca{sub v}β{sub 1a} is a gene transcription regulator. • Here, we show that TnT3 interacts with Ca{sub v}β{sub 1a}. • We mapped TnT3 and Ca{sub v}β{sub 1a} interaction domain. • TnT3 facilitates Ca{sub v}β{sub 1a} nuclear enrichment. • The two proteins play a heretofore unknown role during early muscle differentiation.« less

Synthesis of superconducting Nb 3Sn coatings on Nb substrates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Barzi, E.; Franz, S.; Reginato, F.

In the present work the electrochemical and thermal syntheses of superconductive Nb 3Sn films are investigated. The Nb 3Sn phase is obtained by electrodeposition of Sn layers and Cu intermediate layers onto Nb substrates followed by high temperature diffusion in inert atmosphere. Electrodeposition was performed from aqueous solutions at current densities in the 20 to 50 mA/cm 2 range and at temperatures between 40 and 50°C. Subsequent thermal treatments were realized to obtain the Nb 3Sn superconductive phase. Glow discharge optical emission spectrometry (GDOES) demonstrated that after thermal treatment interdiffusion of Nb and Sn occurred across a thickness of aboutmore » 13 μm. Scanning Electron Microscopy (SEM) allowed accurately measuring the thickness of the Nb 3Sn phase, whose average for the various types of film samples was between 5.7 and 8.0 μm. X-ray diffraction (XRD) patterns confirmed the presence of a cubic Nb 3Sn phase (A15 structure) having (210) preferred orientation. The maximum obtained T c was 17.68 K and the B c20 ranged between 22.5 T and 23.8 T. With the procedure described in the present paper, coating complex shapes cost-effectively becomes possible, which is typical of electrochemical techniques. Furthermore, this approach can be implemented in classical wire processes such as "Jelly Roll" or "Rod in Tube", or directly used for producing superconducting surfaces. In conclusion, the potential of this method for Superconducting Radiofrequency (SRF) structures is also outlined.« less

Synthesis of superconducting Nb 3Sn coatings on Nb substrates

DOE PAGES

Barzi, E.; Franz, S.; Reginato, F.; ...

2015-12-01

In the present work the electrochemical and thermal syntheses of superconductive Nb 3Sn films are investigated. The Nb 3Sn phase is obtained by electrodeposition of Sn layers and Cu intermediate layers onto Nb substrates followed by high temperature diffusion in inert atmosphere. Electrodeposition was performed from aqueous solutions at current densities in the 20 to 50 mA/cm 2 range and at temperatures between 40 and 50°C. Subsequent thermal treatments were realized to obtain the Nb 3Sn superconductive phase. Glow discharge optical emission spectrometry (GDOES) demonstrated that after thermal treatment interdiffusion of Nb and Sn occurred across a thickness of aboutmore » 13 μm. Scanning Electron Microscopy (SEM) allowed accurately measuring the thickness of the Nb 3Sn phase, whose average for the various types of film samples was between 5.7 and 8.0 μm. X-ray diffraction (XRD) patterns confirmed the presence of a cubic Nb 3Sn phase (A15 structure) having (210) preferred orientation. The maximum obtained T c was 17.68 K and the B c20 ranged between 22.5 T and 23.8 T. With the procedure described in the present paper, coating complex shapes cost-effectively becomes possible, which is typical of electrochemical techniques. Furthermore, this approach can be implemented in classical wire processes such as "Jelly Roll" or "Rod in Tube", or directly used for producing superconducting surfaces. In conclusion, the potential of this method for Superconducting Radiofrequency (SRF) structures is also outlined.« less

Biocatalytic route to C-3'-azido/-hydroxy-C-4'-spiro-oxetanoribonucleosides.

PubMed

Kumar, Manish; Sharma, Vivek K; Kumar, Rajesh; Prasad, Ashok K

2015-11-19

The lipase, Novozyme(®)-435, exclusively deacetylates the 5-O-acetyl over 4-C-acetyloxymethyl group of almost identical reactivity in 5-O-acetyl-4-C-acetyloxymethyl-3-azido-3-deoxy-1,2-O-isopropylidene-α-D-ribofuranose that led to the development of first and efficient synthesis of 3'-azido-/3'-amino-C-4'-spiro-oxetanoribonucleosides T, U, C and A in 20-24% overall yields. The X-ray study on the compound obtained by tosylation of lipase-mediated monodeacetylated product unambiguously confirmed the point of diastereoselective monodeacetylation on diacetoxy-azido-ribofuranose derivative. The capability of Novozyme(®)-435 for selective deacylation of 5-O-acetyl group in 5-O-acetyl-4-C-acetyloxymethyl-3-O-benzyl-1,2-O-isopropylidene-α-D-ribofuranose recently discovered by us has been successfully used for the synthesis of C-4'-spiro-oxetanoribonucleosides A and C in good yields. These results clearly indicate that the broader substrate specificity and highly selective capability of Novozyme(®)-435 for carrying out acetylation/deacetylation reactions can be utilized for the development of environment friendly selective methodologies in organic synthesis. Copyright © 2015 Elsevier Ltd. All rights reserved.

tRNAGlu increases the affinity of glutamyl-tRNA synthetase for its inhibitor glutamyl-sulfamoyl-adenosine, an analogue of the aminoacylation reaction intermediate glutamyl-AMP: mechanistic and evolutionary implications.

PubMed

Blais, Sébastien P; Kornblatt, Jack A; Barbeau, Xavier; Bonnaure, Guillaume; Lagüe, Patrick; Chênevert, Robert; Lapointe, Jacques

2015-01-01

For tRNA-dependent protein biosynthesis, amino acids are first activated by aminoacyl-tRNA synthetases (aaRSs) yielding the reaction intermediates aminoacyl-AMP (aa-AMP). Stable analogues of aa-AMP, such as aminoacyl-sulfamoyl-adenosines, inhibit their cognate aaRSs. Glutamyl-sulfamoyl-adenosine (Glu-AMS) is the best known inhibitor of Escherichia coli glutamyl-tRNA synthetase (GluRS). Thermodynamic parameters of the interactions between Glu-AMS and E. coli GluRS were measured in the presence and in the absence of tRNA by isothermal titration microcalorimetry. A significant entropic contribution for the interactions between Glu-AMS and GluRS in the absence of tRNA or in the presence of the cognate tRNAGlu or of the non-cognate tRNAPhe is indicated by the negative values of -TΔSb, and by the negative value of ΔCp. On the other hand, the large negative enthalpy is the dominant contribution to ΔGb in the absence of tRNA. The affinity of GluRS for Glu-AMS is not altered in the presence of the non-cognate tRNAPhe, but the dissociation constant Kd is decreased 50-fold in the presence of tRNAGlu; this result is consistent with molecular dynamics results indicating the presence of an H-bond between Glu-AMS and the 3'-OH oxygen of the 3'-terminal ribose of tRNAGlu in the Glu-AMS•GluRS•tRNAGlu complex. Glu-AMS being a very close structural analogue of Glu-AMP, its weak binding to free GluRS suggests that the unstable Glu-AMP reaction intermediate binds weakly to GluRS; these results could explain why all the known GluRSs evolved to activate glutamate only in the presence of tRNAGlu, the coupling of glutamate activation to its transfer to tRNA preventing unproductive cleavage of ATP.

tRNAGlu Increases the Affinity of Glutamyl-tRNA Synthetase for Its Inhibitor Glutamyl-Sulfamoyl-Adenosine, an Analogue of the Aminoacylation Reaction Intermediate Glutamyl-AMP: Mechanistic and Evolutionary Implications

PubMed Central

Blais, Sébastien P.; Kornblatt, Jack A.; Barbeau, Xavier; Bonnaure, Guillaume; Lagüe, Patrick; Chênevert, Robert; Lapointe, Jacques

2015-01-01

For tRNA-dependent protein biosynthesis, amino acids are first activated by aminoacyl-tRNA synthetases (aaRSs) yielding the reaction intermediates aminoacyl-AMP (aa-AMP). Stable analogues of aa-AMP, such as aminoacyl-sulfamoyl-adenosines, inhibit their cognate aaRSs. Glutamyl-sulfamoyl-adenosine (Glu-AMS) is the best known inhibitor of Escherichia coli glutamyl-tRNA synthetase (GluRS). Thermodynamic parameters of the interactions between Glu-AMS and E. coli GluRS were measured in the presence and in the absence of tRNA by isothermal titration microcalorimetry. A significant entropic contribution for the interactions between Glu-AMS and GluRS in the absence of tRNA or in the presence of the cognate tRNAGlu or of the non-cognate tRNAPhe is indicated by the negative values of –TΔSb, and by the negative value of ΔCp. On the other hand, the large negative enthalpy is the dominant contribution to ΔGb in the absence of tRNA. The affinity of GluRS for Glu-AMS is not altered in the presence of the non-cognate tRNAPhe, but the dissociation constant K d is decreased 50-fold in the presence of tRNAGlu; this result is consistent with molecular dynamics results indicating the presence of an H-bond between Glu-AMS and the 3’-OH oxygen of the 3’-terminal ribose of tRNAGlu in the Glu-AMS•GluRS•tRNAGlu complex. Glu-AMS being a very close structural analogue of Glu-AMP, its weak binding to free GluRS suggests that the unstable Glu-AMP reaction intermediate binds weakly to GluRS; these results could explain why all the known GluRSs evolved to activate glutamate only in the presence of tRNAGlu, the coupling of glutamate activation to its transfer to tRNA preventing unproductive cleavage of ATP. PMID:25860020

The Preferred Substrates for Transglutaminase 2 in a Complex Wheat Gluten Digest Are Peptide Fragments Harboring Celiac Disease T-Cell Epitopes

PubMed Central

Dørum, Siri; Arntzen, Magnus Ø.; Qiao, Shuo-Wang; Holm, Anders; Koehler, Christian J.; Thiede, Bernd; Sollid, Ludvig M.; Fleckenstein, Burkhard

2010-01-01

Background Celiac disease is a T-cell mediated chronic inflammatory disorder of the gut that is induced by dietary exposure to gluten proteins. CD4+ T cells of the intestinal lesion recognize gluten peptides in the context of HLA-DQ2.5 or HLA-DQ8 and the gluten derived peptides become better T-cell antigens after deamidation catalyzed by the enzyme transglutaminase 2 (TG2). In this study we aimed to identify the preferred peptide substrates of TG2 in a heterogeneous proteolytic digest of whole wheat gluten. Methods A method was established to enrich for preferred TG2 substrates in a complex gluten peptide mixture by tagging with 5-biotinamido-pentylamine. Tagged peptides were isolated and then identified by nano-liquid chromatography online-coupled to tandem mass spectrometry, database searching and final manual data validation. Results We identified 31 different peptides as preferred substrates of TG2. Strikingly, the majority of these peptides were harboring known gluten T-cell epitopes. Five TG2 peptide substrates that were predicted to bind to HLA-DQ2.5 did not contain previously characterized sequences of T-cell epitopes. Two of these peptides elicited T-cell responses when tested for recognition by intestinal T-cell lines of celiac disease patients, and thus they contain novel candidate T-cell epitopes. We also found that the intact 9mer core sequences of the respective epitopes were not present in all peptide substrates. Interestingly, those epitopes that were represented by intact forms were frequently recognized by T cells in celiac disease patients, whereas those that were present in truncated versions were infrequently recognized. Conclusion TG2 as well as gastrointestinal proteolysis play important roles in the selection of gluten T-cell epitopes in celiac disease. PMID:21124911

Catalytic roles of the AMP at the 3' end of tRNAs

NASA Technical Reports Server (NTRS)

Wickramasinghe, N. S.; Lacey, J. C. Jr; Lacey JC, J. r. (Principal Investigator)

1994-01-01

Recent reports suggest that the ribosome retains considerable peptidyl transferase activity even when much of the protein of the ribosome is removed and further suggests that rRNA may be the peptidyl transferase. The work here suggests that the AMP residue at the 3' terminus of each tRNA has some catalytic activity both in the esterification reaction and in forming a pseudopeptide, AcGly, and further suggests that whatever peptidyl transferase is, it finds a cooperative substrate in the aminoacyl-AMP at the 3' terminus of tRNA.

Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane

PubMed Central

2017-01-01

The biological potential of Vip and Cry proteins from Bacillus is well known and widely established. Thus, it is important to look for new genes showing different modes of action, selecting those with differentiated entomotoxic activity against Diatraea flavipennella and Elasmopalpus lignosellus, which are secondary pests of sugarcane. Therefore, Cry1 and Vip3 proteins were expressed in Escherichia coli, and their toxicities were evaluated based on bioassays using neonate larvae. Of those, the most toxic were Cry1Ac and Vip3Aa considering the LC50 values. Toxins from E. coli were purified, solubilized, trypsinized, and biotinylated. Brush Border Membrane Vesicles (BBMVs) were prepared from intestines of the two species to perform homologous and heterologous competition assays. The binding assays demonstrated interactions between Cry1Aa, Cry1Ac, and Vip3Aa toxins and proteins from the BBMV of D. flavipennella and E. lignosellus. Homologous competition assays demonstrated that binding to one of the BBMV proteins was specific for each toxin. Heterologous competition assays indicated that Vip3Aa was unable to compete for Cry1Ac toxin binding. Our results suggest that Cry1Ac and Vip3Aa may have potential in future production of transgenic sugarcane for control of D. flavipennella and E. lignosellus, but more research is needed on the potential antagonism or synergism of the toxins in these pests. PMID:28123906

The yeast rapid tRNA decay pathway competes with elongation factor 1A for substrate tRNAs and acts on tRNAs lacking one or more of several modifications.

PubMed

Dewe, Joshua M; Whipple, Joseph M; Chernyakov, Irina; Jaramillo, Laura N; Phizicky, Eric M

2012-10-01

The structural and functional integrity of tRNA is crucial for translation. In the yeast Saccharomyces cerevisiae, certain aberrant pre-tRNA species are subject to nuclear surveillance, leading to 3' exonucleolytic degradation, and certain mature tRNA species are subject to rapid tRNA decay (RTD) if they are appropriately hypomodified or bear specific destabilizing mutations, leading to 5'-3' exonucleolytic degradation by Rat1 and Xrn1. Thus, trm8-Δ trm4-Δ strains are temperature sensitive due to lack of m(7)G(46) and m(5)C and the consequent RTD of tRNA(Val(AAC)), and tan1-Δ trm44-Δ strains are temperature sensitive due to lack of ac(4)C(12) and Um(44) and the consequent RTD of tRNA(Ser(CGA)) and tRNA(Ser(UGA)). It is unknown how the RTD pathway interacts with translation and other cellular processes, and how generally this pathway acts on hypomodified tRNAs. We provide evidence here that elongation factor 1A (EF-1A) competes with the RTD pathway for substrate tRNAs, since its overexpression suppresses the tRNA degradation and the growth defect of strains subject to RTD, whereas reduced levels of EF-1A have the opposite effect. We also provide evidence that RTD acts on a variety of tRNAs lacking one or more different modifications, since trm1-Δ trm4-Δ mutants are subject to RTD of tRNA(Ser(CGA)) and tRNA(Ser(UGA)) due to lack of m(2,2)G(26) and m(5)C, and since trm8-Δ, tan1-Δ, and trm1-Δ single mutants are each subject to RTD. These results demonstrate that RTD interacts with the translation machinery and acts widely on hypomodified tRNAs.

Preparation of epitaxial TlBa2Ca2Cu3O9 high Tc thin films on LaAlO3 (100) substrates

NASA Astrophysics Data System (ADS)

Piehler, A.; Reschauer, N.; Spreitzer, U.; Ströbel, J. P.; Schönberger, R.; Renk, K. F.; Saemann-Ischenko, G.

1994-09-01

Epitaxial TlBa2Ca2Cu3O9 high Tc thin films were prepared on LaAlO3 (100) substrates by a combination of laser ablation and thermal evaporation of thallium oxide. X-ray diffraction patterns of θ-2θ scans showed that the films consisted of highly c axis oriented TlBa2Ca2Cu3O9. φ scan measurements revealed an epitaxial growth of the TlBa2Ca2Cu3O9 thin films on the LaAlO3 (100) substrates. Ac inductive measurements indicated the onset of superconductivity at 110 K. At 6 K, the critical current density was 4×106 A/cm2 in zero magnetic field and 6×105 A/cm2 at a magnetic field of 3 T parallel to the c axis.

The T-cell antigen CD5 acts as a receptor and substrate for the protein-tyrosine kinase p56lck.

PubMed Central

Raab, M; Yamamoto, M; Rudd, C E

1994-01-01

CD5 is a T-cell-specific antigen which binds to the B-cell antigen CD72 and acts as a coreceptor in the stimulation of T-cell growth. CD5 associates with the T-cell receptor zeta chain (TcR zeta)/CD3 complex and is rapidly phosphosphorylated on tyrosine residues as a result of TcR zeta/CD3 ligation. However, despite this, the mechanism by which CD5 generates intracellular signals is unclear. In this study, we demonstrate that CD5 is coupled to the protein-tyrosine kinase p56lck and can act as a substrate for p56lck. Coexpression of CD5 with p56lck in the baculovirus expression system resulted in the phosphorylation of CD5 on tyrosine residues. Further, anti-CD5 and anti-p56lck coprecipitated each other in a variety of detergents, including Nonidet P-40 and Triton X-100. Anti-CD5 also precipitated the kinase from various T cells irrespective of the expression of TcR zeta/CD3 or CD4. No binding between p59fyn(T) and CD5 was detected in T cells. The binding of p56lck to CD5 induced a 10- to 15-fold increase in p56lck catalytic activity, as measured by in vitro kinase analysis. In vivo labelling with 32P(i) also showed a four- to fivefold increase in Y-394 occupancy in p56lck when associated with CD5. The use of glutathione S-transferase-Lck fusion proteins in precipitation analysis showed that the SH2 domain of p56lck could recognize CD5 as expressed in the baculovirus expression system. CD5 interaction with p56lck represents a novel variant of a receptor-kinase complex in which receptor can also serve as substrate. The CD5-p56lck interaction is likely to play roles in T-cell signalling and T-B collaboration. Images PMID:7513045

The growth of small corrosion fatigue cracks in alloy 2024

NASA Technical Reports Server (NTRS)

Piascik, Robert S.; Willard, Scott A.

1993-01-01

The corrosion fatigue crack growth characteristics of small surface and corner cracks in aluminum alloy 2024 is established. The damaging effect of salt water on the early stages of small crack growth is characterized by crack initiation at constituent particle pits, intergranular microcracking for a less than 100 micrometers, and transgranular small crack growth for a micrometer. In aqueous 1 percent NaCl and at a constant anodic potential of -700 mV(sub SCE), small cracks exhibit a factor of three increase in fatigue crack growth rates compared to laboratory air. Small cracks exhibit accelerated corrosion fatigue crack growth rates at low levels of delta-K (less than 1 MPa square root of m) below long crack delta-K (sub th). When exposed to Paris regime levels of crack tip stress intensity, small corrosion fatigue cracks exhibit growth rates similar to that observed for long cracks. Results suggest that crack closure effects influence the corrosion fatigue crack growth rates of small cracks (a less than or equal to 100 micrometers). This is evidenced by similar small and long crack growth behavior at various levels of R. Contrary to the corrosion fatigue characteristics of small cracks in high strength steels, no pronounced chemical crack length effect is observed for Al by 2024 exposed to salt water.

Constitutive behavior of as-cast AA1050, AA3104, and AA5182

NASA Astrophysics Data System (ADS)

van Haaften, W. M.; Magnin, B.; Kool, W. H.; Katgerman, L.

2002-07-01

Recent thermomechanical modeling to calculate the stress field in industrially direct-chill (DC) cast-aluminum slabs has been successful, but lack of material data limits the accuracy of these calculations. Therefore, the constitutive behavior of three aluminum alloys (AA1050, AA3104, and AA5182) was determined in the as-cast condition using tensile tests at low strain rates and from room temperature to solidus temperature. The parameters of two constitutive equations, the extended Ludwik equation and a combination of the Sellars-Tegart equation with a hardening law, were determined. In order to study the effect of recovery, the constitutive behavior after prestraining at higher temperatures was also investigated. To evaluate the quantified constitutive equations, tensile tests were performed simulating the deformation and cooling history experienced by the material during casting. It is concluded that both constitutive equations perform well, but the combined hardening-Sellars-Tegart (HST) equation has temperature-independent parameters, which makes it easier to implement in a DC casting model. Further, the deformation history of the ingot should be taken into account for accurate stress calculations.

Analysing the Friction Stir Welded Joints of AA2219 Al-Cu Alloy in Different Heat-Treated-State

NASA Astrophysics Data System (ADS)

Venkateswarlu, D.; Cheepu, Muralimohan; Kranthi kumar, B.; Mahapatra, M. M.

2018-03-01

Aluminium alloy AA2219 is widely used in light weight structural applications where the good corrosion resistance and specific weight required. The fabrication of this alloy using friction stir welding process is gaining interest towards finding the characteristics of the weld metal properties, since this process involved in the welded materials does not melt and recast. In the present investigation, friction stir welding process was used for different heat treated conditions of 2219-T87 and 2219-T62 aluminium alloys to find the influence of base metal on characteristics of the joints. The experimental output results exhibited that, mechanical properties, weld metal characteristics and joint failure locations are significantly affected by the different heat treatment conditions of the substrate. The joints tensile and yield strength of the 2219-T87 welds was higher than the 2219-T62 welds. Hardness distribution in the stir zone was significantly varied between two different heat treaded material conditions. The microstructural features of the 2219-T62 welds reveal the coarse grains formation in the thermo-mechanically affected zone and heat affected zone. The joint efficiency of the 2219- T82 welds is 59.87%, while that of 2219-T62 welds is 39.10%. In addition, the elongation of the joint also varied and the joints failure location characteristics are different for two different types heat treated condition joints.

Trends in diabetes incidence from 1992 to 2015 and projections for 2024: A Portuguese General Practitioner's Network study.

PubMed

de Sousa-Uva, Mafalda; Antunes, L; Nunes, B; Rodrigues, A P; Simões, J A; Ribeiro, R T; Boavida, J M; Matias-Dias, C

2016-10-01

Diabetes is known as a major cause of morbidity and mortality worldwide. Portugal is known as the European country with the highest prevalence of this disease. While diabetes prevalence data is updated annually in Portugal, the General Practitioner's (GP) Sentinel Network represents the only data source on diabetes incidence. This study describes the trends in Diabetes incidence, between 1992 and 2015, and estimate projections for the future incidence rates in Portugal until 2024. An ecological time-series study was conducted using data from GP Sentinel Network between 1992 and 2015. Family doctors reported all new cases of Diabetes in their patients' lists. Annual trends were estimated through Poisson regression models as well as the future incidence rates (until 2024), sex and age group stratified. Incidence rate projections were adjusted to the distribution of the resident Portuguese population given Statistics Portugal projections. The average increase in Diabetes incidence rate was in total 4.29% (CI95% 3.80-4.80) per year under study. Until 1998-2000, the annual incidence rate was higher in women, and from 1998-2000 to 2013-2015 turn out to be higher in men. The incidence rate projected for 2022-2024 was 972.77/10(5) inhabitants in total, and 846.74/10(5) and 1114.42/10(5), respectively, in women and men. This is the first study in Portugal to estimate diabetes incidence rate projections. The disturbing reported projections seem realistic if things continue as in the past. Actually, effective public health policies will need to be undertaken to minimize this alarming future scenario. Copyright © 2016 Primary Care Diabetes Europe. Published by Elsevier Ltd. All rights reserved.

Effect of Multi-Pass Friction Stir Processing on Mechanical Properties for AA2024/Al2O3 Nanocomposites

PubMed Central

2017-01-01

In the present work, an aluminum metal matrix reinforced with (Al2O3) nanoparticles was fabricated as a surface composite sheet using friction stir processing (FSP). The effects of processing parameters on mechanical properties, hardness, and microstructure grain were investigated. The results revealed that multi-pass FSP causes a homogeneous distribution and good dispersion of Al2O3 in the metal matrix, and consequently an increase in the hardness of the matrix composites. A finer grain is observed in the microstructure examination in specimens subjected to second and third passes of FSP. The improvement in the grain refinement is 80% compared to base metal. The processing parameters, particularly rotational tool speed and pass number in FSP, have a major effect on strength properties and surface hardness. The ultimate tensile strength (UTS) and the average hardness are improved by 25% and 46%, respectively, due to presence of reinforcement Al2O3 nanoparticles. PMID:28885575

Effect of Scandium on the Interaction of Concurrent Precipitation and Recrystallization in Commercial AA3003 Aluminum Alloy

NASA Astrophysics Data System (ADS)

Tu, Yiyou; Qian, Huan; Zhou, Xuefeng; Jiang, Jianqing

2014-04-01

In the current study, the effect of Sc addition on the interaction of concurrent precipitation and recrystallization in commercial AA3003 aluminum alloy was investigated using optical microscopy, scanning electron microscopy, and transmission electron microscopy. In case of AA3003 alloy, which was cold rolled to a true strain of 2.20 and heated at a heating rate of 150 K/s, the onset of precipitation and ending of recrystallization are signified by the critical temperature, T C ~740 K (467 °C). There is a change in the shape of the recrystallized grains from pancake-like to equiaxed shape, as the annealing temperature increases greater than T C. In case of AA3003 alloy microalloyed with 0.4 wt pct of Sc, the high no. density precipitation of coherent Al3Sc precipitates always occurs before recrystallization because of the small nucleation barrier and high rate of decomposition. This leads to extremely coarse pancake-like recrystallization grains with high fraction of low-angle grain boundaries in the entire annealing temperature range, even at a high brazing temperature of 883 K (610 °C).

Sipuleucel-T: Prototype for development of anti-tumor vaccines.

PubMed

Carballido, Estrella; Fishman, Mayer

2011-04-01

Prostate cancer immunotherapy officially debuted with the recent FDA approval of Sipuleucel-T. The novel trend of cancer immunotherapy relies on the identification of particular tumor-associated antigens, like prostatic acid phosphatase (PAP). Sipuleucel-T consists of autologous dendritic cells activated in vitro with recombinant fusion protein PA2024, PAP-linked to granulocyte-macrophage colony-stimulating factor. Sipuleucel-T represents a prototype for the development of cancer vaccines. Preclinical and clinical data as well as landmark studies for the existing narrow chemotherapy alternatives and early immunotherapy trials will be discussed. The pivotal trial demonstrated a 4.1-month difference of median survival, but with no effect on time to progression in asymptomatic or minimally symptomatic metastatic castrate-resistant patients. Several immunologic effects were observed in the treated population, including antibody and T cell-specific activity to P2024 and PAP. With all new therapies the extent of clinical and objective benefits versus encountered limitations should be evaluated. This review highlights the events and decisions in the process of the development of Sipuleucel-T. We discuss how this successful immunotherapy outcome challenges us to use it as a starting point for variations to or try to amplify practical anticancer progress within the antitumor vaccine paradigm.

AAS 228: Day 3 morning

NASA Astrophysics Data System (ADS)

Kohler, Susanna

2016-06-01

Editors Note:This week were at the 228th AAS Meeting in San Diego, CA. Along with a team ofauthors from astrobites.com, I will bewritingupdates on selectedevents at themeeting and posting twiceeach day. Follow along here or atastrobites.com, or catch ourlive-tweeted updates from the@astrobites Twitter account. The usual posting schedule for AAS Nova will resumenext week.Plenary Session 2015 Newton Lacy Pierce Prize Lecture: The Elephant in the Room: Effects of Distant, Massive Companions on Planetary System Architectures (by Leonardo dos Santos)The first session on Wednesday at 228th AAS Meeting was the Newton Lacy Pierce Prize Lecture by Heather Knutson (California Institute of Technology). This talk featured a broad range of research efforts on exoplanets, with the main focus on how we study the composition of their atmospheres, and how multi-body interactions carve the structure of the planetary systems we observe.One of her first points is the well-known idea that the Solar System is an oddball, compared to the exoplanet systems we have found so far: most of these systems contain hot Jupiters and mini-Neptunes at very close-in orbits around their host stars. Moreover, even when studying their transmission spectra, it is difficult to know the exact composition of their atmospheres.Knutson: it is difficult to constrain atmospheric composition of exoplanets (H-poor or H-rich+clouds?) #aas228pic.twitter.com/LdyN4o9RC7 astrobites (@astrobites) June 15, 2016The main proposal on how these systems formed is the migration scenario. In order to validate this idea, Dr. Knutson and her group The Friends of Hot Jupiters study systems with close-in gas giants and their frequency of binary companions, which are supposed to be the main culprits causing gas-giant migration. They found that approximately half of the observed systems have long-distance companions, providing strong validation of the migration scenario. Moreover, Dr. Knutson speculates that wide binaries have more

Structure of RNA 3′-phosphate cyclase bound to substrate RNA

PubMed Central

Desai, Kevin K.; Bingman, Craig A.; Cheng, Chin L.; Phillips, George N.

2014-01-01

RNA 3′-phosphate cyclase (RtcA) catalyzes the ATP-dependent cyclization of a 3′-phosphate to form a 2′,3′-cyclic phosphate at RNA termini. Cyclization proceeds through RtcA–AMP and RNA(3′)pp(5′)A covalent intermediates, which are analogous to intermediates formed during catalysis by the tRNA ligase RtcB. Here we present a crystal structure of Pyrococcus horikoshii RtcA in complex with a 3′-phosphate terminated RNA and adenosine in the AMP-binding pocket. Our data reveal that RtcA recognizes substrate RNA by ensuring that the terminal 3′-phosphate makes a large contribution to RNA binding. Furthermore, the RNA 3′-phosphate is poised for in-line attack on the P–N bond that links the phosphorous atom of AMP to Nε of His307. Thus, we provide the first insights into RNA 3′-phosphate termini recognition and the mechanism of 3′-phosphate activation by an Rtc enzyme. PMID:25161314

Structural and functional importance of transmembrane domain 3 (TM3) in the aspartate:alanine antiporter AspT: topology and function of the residues of TM3 and oligomerization of AspT.

PubMed

Nanatani, Kei; Maloney, Peter C; Abe, Keietsu

2009-04-01

AspT, the aspartate:alanine antiporter of Tetragenococcus halophilus, a membrane protein of 543 amino acids with 10 putative transmembrane (TM) helices, is the prototype of the aspartate:alanine exchanger (AAE) family of transporters. Because TM3 (isoleucine 64 to methionine 85) has many amino acid residues that are conserved among members of the AAE family and because TM3 contains two charged residues and four polar residues, it is thought to be located near (or to form part of) the substrate translocation pathway that includes the binding site for the substrates. To elucidate the role of TM3 in the transport process, we carried out cysteine-scanning mutagenesis. The substitutions of tyrosine 75 and serine 84 had the strongest inhibitory effects on transport (initial rates of l-aspartate transport were below 15% of the rate for cysteine-less AspT). Considerable but less-marked effects were observed upon the replacement of methionine 70, phenylalanine 71, glycine 74, arginine 76, serine 83, and methionine 85 (initial rates between 15% and 30% of the rate for cysteine-less AspT). Introduced cysteine residues at the cytoplasmic half of TM3 could be labeled with Oregon green maleimide (OGM), whereas cysteines close to the periplasmic half (residues 64 to 75) were not labeled. These results suggest that TM3 has a hydrophobic core on the periplasmic half and that hydrophilic residues on the cytoplasmic half of TM3 participate in the formation of an aqueous cavity in membranes. Furthermore, the presence of l-aspartate protected the cysteine introduced at glycine 62 against a reaction with OGM. In contrast, l-aspartate stimulated the reactivity of the cysteine introduced at proline 79 with OGM. These results demonstrate that TM3 undergoes l-aspartate-induced conformational alterations. In addition, nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses and a glutaraldehyde cross-linking assay suggest that functional AspT forms homo-oligomers as a

Mutation of the human mitochondrial phenylalanine-tRNA synthetase causes infantile-onset epilepsy and cytochrome c oxidase deficiency.

PubMed

Almalki, Abdulraheem; Alston, Charlotte L; Parker, Alasdair; Simonic, Ingrid; Mehta, Sarju G; He, Langping; Reza, Mojgan; Oliveira, Jorge M A; Lightowlers, Robert N; McFarland, Robert; Taylor, Robert W; Chrzanowska-Lightowlers, Zofia M A

2014-01-01

Mitochondrial aminoacyl-tRNA synthetases (aaRSs) are essential enzymes in protein synthesis since they charge tRNAs with their cognate amino acids. Mutations in the genes encoding mitochondrial aaRSs have been associated with a wide spectrum of human mitochondrial diseases. Here we report the identification of pathogenic mutations (a partial genomic deletion and a highly conserved p. Asp325Tyr missense variant) in FARS2, the gene encoding mitochondrial phenylalanyl-tRNA synthetase, in a patient with early-onset epilepsy and isolated complex IV deficiency in muscle. The biochemical defect was expressed in myoblasts but not in fibroblasts and associated with decreased steady state levels of COXI and COXII protein and reduced steady state levels of the mt-tRNA(Phe) transcript. Functional analysis of the recombinant mutant p. Asp325Tyr FARS2 protein showed an inability to bind ATP and consequently undetectable aminoacylation activity using either bacterial tRNA or human mt-tRNA(Phe) as substrates. Lentiviral transduction of cells with wildtype FARS2 restored complex IV protein levels, confirming that the p.Asp325Tyr mutation is pathogenic, causing respiratory chain deficiency and neurological deficits on account of defective aminoacylation of mt-tRNA(Phe). © 2013. Published by Elsevier B.V. All rights reserved.

Role of Ca2+-independent phospholipase A2 and cytochrome P-450 in store-operated calcium entry in 3T6 fibroblasts.

PubMed

Martínez, Javier; Moreno, Juan J

2005-09-01

Store-operated calcium (SOC) channels and capacitative Ca2+ entry play a key role in cellular functions, but their mechanism of activation remains unclear. Here, we show that thapsigargin induces [3H] arachidonic acid (AA) release, 45Ca2+ influx and a subsequent enhancement of intracellular calcium concentration ([Ca2+]i. Thapsigargin-induced elevation of [Ca2+]i was inhibited by cytochrome P-450 inhibitors and by cytochrome P-450 epoxygenase inhibitor and was reverted by 11,12 EET addition. However, cyclooxygenase and lipoxygenase inhibitors have no effect. Moreover, we observed that four EETs were able to induce 45Ca2+ influx. Finally, we reported that the effect of 11,12 EET on 45Ca2+ influx was sensible to receptor-operated Ca2+ channel blockers (NiCl2, LaCl3) but not to voltage-dependent Ca2+ channel blocker as verapamil. Thus, AA released by Ca2+-independent phospholipase A2 and AA metabolism through cytochrome P-450 pathway may be crucial molecular determinant in thapsigargin activation of SOC channels and store-operated Ca2+ entry pathway in 3T6 fibroblasts. Moreover, EETs, the main cytochrome P-450 epoxygenase metabolites of AA, are involved in thapsigargin-stimulated Ca2+ influx. In summary, our results suggest that EETs are components of calcium influx factor(s).

High-Speed Friction Stir Welding of AA7075-T6 Sheet: Microstructure, Mechanical Properties, Micro-texture, and Thermal History

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Jingyi; Upadhyay, Piyush; Hovanski, Yuri

Friction-stir-welding (FSW) is a cost-effective and high quality joining process for aluminum alloys (especially heat-treatable allo ys) that has been applied successfully in the aerospace industry. However, the full potential of FSW on more cost-sensitive applications is still limited by the production rate, namely the welding speed of the process. The majority of literature evaluating FSW of aluminum alloys is based on welds made in the range of welding speeds around hundreds of millimeters per minute, and only a handful are at a moderate speed of 1 m/min. In this study we present a microstructural analysis of friction stir weldedmore » AA7075-T6 blanks with welding speeds up to 3 m/min. Textures, microstructures, mechanical properties, and weld quality are analyzed using TEM, EBSD, metallographic imaging, and Vickers hardness. Results are coupled with welding parameters to aid in the understanding of the complex material flow and texture gradients within the welds in an effort to optimize welding parameters for high speed processing.« less

Long range wetting transparency on top of layered metal dielectric substrates

DTIC Science & Technology

2015-11-20

multi-layered stacks were deposited onto glass substrates ( silica -based Micro cover glass , 22mmx22mm from VWR (48366-067), index of refraction n...necessarily endorsed by the United States Government. Long-range wetting transparency on top of layered metal-dielectric substrates M. A...as far as ~100 nm beneath the water/MgF2 interface. We refer to this phenomenon as long range wetting transparency . The latter effect cannot be

Bacillus thuringiensis Cry3Aa toxin increases the susceptibility of Crioceris quatuordecimpunctata to Beauveria bassiana infection

USDA-ARS?s Scientific Manuscript database

The spotted asparagus beetle, Crioceris quatuordecimpunctata (Coleoptera: Chrysomelidae), is one of the most devastating pests of asparagus in China and elsewhere. In this study, we investigated the interaction of Bacillus thuringiensis (Bt) Cry3Aa toxin and the entomopathogenic fungus Beauveria bas...

Longitudinal study of experimental induction of AA amyloidosis in mice seeded with homologous and heterologous AA fibrils.

PubMed

Muhammad, Naeem; Murakami, Tomoaki; Inoshima, Yasuo; Ishiguro, Naotaka

2016-09-01

To investigate pathogenesis and kinetics of experimentally induced murine AA amyloidosis seeded with homologous (murine) and heterologous (bovine) AA fibrils. Experimental AA amyloidosis was induced by administration of inflammatory stimulus and preformed AA fibrils to a total of 111 female C57/Black mice. In this longitudinal study, heterologous (bovine) as well as homologous (murine) AA fibrils were injected intraperitoneally to mice in various combinations. Re-stimulation was done at 120 or 300 days post first inoculation. To analyze the intensity of amyloid depositions in mice organs, immunohistochemical techniques and image J software were used. Assessment of cytokines level in sera was done using a Mouse Th1/Th2/Th17 Cytokine CBA Kit. Incidence and severity of AA amyloidosis were quite low in mice inoculated with heterologous bovine AA fibrils than homologous murine one. Homologous AA fibrils administration at first and second inoculation caused maximum amount of amyloid depositions and severe systemic form of amyloidosis. Increase in the level of pro-inflammatory cytokine IL-6 was observed after first inoculation, while second inoculation caused a further increase in the level of anti-inflammatory cytokine IL-10. AA amyloidosis can be induced by heterologous as well as homologous AA fibrils. Severity of AA amyloidosis induced with homologous AA fibrils is higher compared to heterologous AA fibrils.

Encapsulated oligodendrocyte precursor cell fate is dependent on PDGF-AA release kinetics in a 3D microparticle-hydrogel drug delivery system.

PubMed

Pinezich, Meghan R; Russell, Lauren N; Murphy, Nicholas P; Lampe, Kyle J

2018-04-16

Biomaterial drug delivery systems (DDS) can be used to regulate growth factor release and combat the limited intrinsic regeneration capabilities of central nervous system (CNS) tissue following injury and disease. Of particular interest are systems that aid in oligodendrocyte regeneration, as oligodendrocytes generate myelin which surrounds neuronal axons and helps transmit signals throughout the CNS. Oligodendrocyte precursor cells (OPCs) are found in small numbers in the adult CNS, but are unable to effectively differentiate following CNS injury. Delivery of signaling molecules can initiate a favorable OPC response, such as proliferation or differentiation. Here, we investigate the delivery of one such molecule, platelet derived growth factor-AA (PDGF-AA), from poly(lactic-co-glycolic) acid microparticles to OPCs in a 3D polyethylene glycol-based hydrogel. The goal of this DDS was to better understand the relationship between PDGF-AA release kinetics and OPC fate. The system approximates native brain tissue stiffness, while incorporating PDGF-AA under seven different delivery scenarios. Within this DDS, supply of PDGF-AA followed by PDGF-AA withdrawal caused OPCs to upregulate gene expression of myelin basic protein (MBP) by factors of 1.6-9.2, whereas continuous supply of PDGF-AA caused OPCs to remain proliferative. At the protein expression level, we observed an upregulation in O1, a marker for mature oligodendrocytes. Together, these results show that burst release followed by withdrawal of PDGF-AA from a hydrogel DDS stimulates survival, proliferation, and differentiation of OPCs in vitro. Our results could inform the development of improved neural regeneration strategies that incorporate delivery of PDGF-AA to the injured CNS. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2018. © 2018 Wiley Periodicals, Inc.

SU-E-T-209: Comparison of Plan Quality Between Arm Avoidance (AA) Vs. Non Arm Avoidance VMAT Planning Techniques for Breast Cancer Patients with Bilateral Implant Reconstructions Receiving Postmastectomy Radiation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kuo, L; Ballangrud, A; Ho, A

Purpose: Breast cancer patients with bilateral implant reconstructions who require postmastectomy radiotherapy can pose unique treatment planning challenges. The use of VMAT may provide advantages over conventional tangent or multi-beam IMRT techniques. Moreover, daily setup uncertainly of the arm position, however, could have significant impact on accurate dose delivery. This study compares the plan qualities between non-AA and AA VMAT techniques. Methods: Three breast cancer patients receiving left chest wall and regional nodal irradiation with bilateral implant reconstructions were studied. PTV included chest wall and IMNs (PTV-CW), and supraclavicular and axillary lymph nodes (PTV-SCV). For each patient, one non-AA VMATmore » plan (VMAT-S) with 4 partial arcs encompassing the ipsilateral arm and three AA VMAT plans where no arcs were entering or existing through the ipsilateral arm were generated. VMAT-AA1 uses 2 arcs for PTV-CW and 2 arcs for PTV-SCV. VMAT-AA2 used two static fields for PTV-SCV with 2 arcs for PTV-CW. VMAT-AA3 used 2 narrow arcs for PTV-CW and 2 long arcs for all PTVs. Prescription dose (PD) was 50 Gy (25 fractions). All plans were normalized to have 95% of PD to 95 % of PTV. PTV dose inhomogeneity and dose to the heart, left lung, right thyroid dose and left humerus were evaluated. Results: For VMAT-S, VMAT-AA1, VMAT-AA2 and VMAT-AA3, respectively, the average and standard deviation (in Gy unless specified otherwise) of PTV D05 are 54.7±0.9, 55.9±0.4, 56.7±0.7 and 55.7±0.4; mean Heart dose: 7.1±0.7, 7.2±0.8, 7.3±0.9 and 6.9±1.0; left lung V20Gy (in %): 28.1±1.0, 28.8+2.2, 32.2±4.1 and 27.8±2.0; mean right thyroid dose: 8.1±0.6, 5.1±2.1, 2.1±0.4 and 5.0±2.0; mean left humerus dose: 20.0±4.4,15.6±4.4, 15.2±8.2 and 15.3±4.6. Conclusion: AA VMAT can produce acceptable clinical plans while eliminating dosimetric impact related to arm setup uncertainty. These data require validation in larger planning studies prior to routine

New Insights into the Role of T3 Loop in Determining Catalytic Efficiency of GH28 Endo-Polygalacturonases

PubMed Central

Tu, Tao; Meng, Kun; Luo, Huiying; Turunen, Ossi; Zhang, Lujia; Cheng, Yanli; Su, Xiaoyun; Ma, Rui; Shi, Pengjun; Wang, Yaru; Yang, Peilong; Yao, Bin

2015-01-01

Intramolecular mobility and conformational changes of flexible loops have important roles in the structural and functional integrity of proteins. The Achaetomium sp. Xz8 endo-polygalacturonase (PG8fn) of glycoside hydrolase (GH) family 28 is distinguished for its high catalytic activity (28,000 U/mg). Structure modeling indicated that PG8fn has a flexible T3 loop that folds partly above the substrate in the active site, and forms a hydrogen bond to the substrate by a highly conserved residue Asn94 in the active site cleft. Our research investigates the catalytic roles of Asn94 in T3 loop which is located above the catalytic residues on one side of the substrate. Molecular dynamics simulation performed on the mutant N94A revealed the loss of the hydrogen bond formed by the hydroxyl group at O34 of pentagalacturonic acid and the crucial ND2 of Asn94 and the consequent detachment and rotation of the substrate away from the active site, and that on N94Q caused the substrate to drift away from its place due to the longer side chain. In line with the simulations, site-directed mutagenesis at this site showed that this position is very sensitive to amino acid substitutions. Except for the altered K m values from 0.32 (wild type PG8fn) to 0.75–4.74 mg/ml, all mutants displayed remarkably lowered k cat (~3–20,000 fold) and k cat/K m (~8–187,500 fold) values and significantly increased △(△G) values (5.92–33.47 kJ/mol). Taken together, Asn94 in the GH28 T3 loop has a critical role in positioning the substrate in a correct way close to the catalytic residues. PMID:26327390

Control of Alq3 wetting layer thickness via substrate surface functionalization.

PubMed

Tsoi, Shufen; Szeto, Bryan; Fleischauer, Michael D; Veinot, Jonathan G C; Brett, Michael J

2007-06-05

The effects of substrate surface energy and vapor deposition rate on the initial growth of porous columnar tris(8-hydroxyquinoline)aluminum (Alq3) nanostructures were investigated. Alq3 nanostructures thermally evaporated onto as-supplied Si substrates bearing an oxide were observed to form a solid wetting layer, likely caused by an interfacial energy mismatch between the substrate and Alq3. Wetting layer thickness control is important for potential optoelectronic applications. A dramatic decrease in wetting layer thickness was achieved by depositing Alq3 onto alkyltrichlorosilane-derivatized Si/oxide substrates. Similar effects were noted with increasing deposition rates. These two effects enable tailoring of the wetting layer thickness.

Investigation of decolorization of textile wastewater in an anaerobic/aerobic biological activated carbon system (A/A BAC).

PubMed

Pasukphun, N; Vinitnantharat, S; Gheewala, S

2010-04-01

The aim of this study is to investigate the decolorization in anaerobic/aerobic biological activated carbon (A/A BAC) system. The experiment was divided into 2 stages; stage I is batch test for preliminary study of dye removal equilibrium time. The preliminary experiment (stage I) provided the optimal data for experimental design of A/A BAC system in SBR (stage II). Stage II is A/A BAC system imitated Sequencing Batch Reactor (SBR) which consist of 5 main periods; fill, react, settle, draw and idle. React period include anaerobic phase followed by aerobic phase. The BAC main media; Granular Activated Carbon (GAC), Mixed Cultures (MC) and Biological Activated Carbon (BAC) were used for dye and organic substances removal in three different solutions; Desizing Agent Solution (DAS), dye Solution (DS) and Synthetic Textile Wastewater (STW). Results indicate that GAC adsorption plays role in dye removal followed by BAC and MC activities, respectively. In the presence desizing agent, decolorization by MC was improved because desizing agent acts as co-substrates for microorganisms. It was found that 50% of dye removal efficiency was achieved in Fill period by MC. GC/MS analysis was used to identify dye intermediate from decolorization. Dye intermediate containing amine group was found in the solution and on BAC surfaces. The results demonstrated that combination of MC and BAC in the system promotes decolorization and dye intermediate removal. In order to improve dye removal efficiency in an A/A BAC system, replacement of virgin GAC, sufficient co-substrates supply and the appropriate anaerobic: aerobic period should be considered.

Low-Temperature Friction-Stir Welding of 2024 Aluminum

NASA Technical Reports Server (NTRS)

Benavides, S.; Li, Y.; Murr, L. E.; Brown, D.; McClure, J. C.

1998-01-01

Solid state friction-stir welding (FSW) has been demonstrated to involve dynamic recrystallization producing ultra-fine, equiaxed grain structures to facilitate superplastic deformation as the welding or joining mechanism. However, the average residual, equiaxed, grain size in the weld zone has ranged from roughly 0.5 micron to slightly more than 10 micron, and the larger weld zone grain sizes have been characterized as residual or static grain growth as a consequence of the temperatures in the weld zone (where center-line temperatures in the FSW of 6061 Al have been shown to be as high as 480C or -0.8 T(sub M) where T(sub M) is the absolute melting temperature)). In addition, the average residual weld zone grain size has been observed to increase near the top of the weld, and to decrease with distance on either side of the weld-zone centerline, an d this corresponds roughly to temperature variations within the weld zone. The residual grain size also generally decreases with decreasing FSW tool rotation speed. These observations are consistent with the general rules for recrystallization where the recrystallized grain size decreases with increasing strain (or deformation) at constant strain rate, or with increasing strain-rate, or with increasing strain rate at constant strain; especially at lower ambient temperatures, (or annealing temperatures). Since the recrystallization temperature also decreases with increasing strain rate, the FSW process is somewhat complicated because the ambient temperature, the frictional heating fraction, and the adiabatic heating fraction )proportional to the product of strain and strain-rate) will all influence both the recrystallization and growth within the FSW zone. Significantly reducing the ambient temperature of the base metal or work pieces to be welded would be expected to reduce the residual weld-zone grain size. The practical consequences of this temperature reduction would be the achievement of low temperature welding. This

Simultaneous quantification of T4, T3, rT3, 3,5-T2 and 3,3'-T2 in larval zebrafish (Danio rerio) as a model to study exposure to polychlorinated biphenyls.

PubMed

Chen, Xiaopeng; Walter, Kyla M; Miller, Galen W; Lein, Pamela J; Puschner, Birgit

2018-06-01

Environmental toxicants that interfere with thyroid hormone (TH) signaling can impact growth and development in animals and humans. Zebrafish represent a model to study chemically induced TH disruption, prompting the need for sensitive detection of THs. Simultaneous quantification of 3,3',5-triiodo-l-thyronine (T3), thyroxine (T4), 3,3',5'-triiodo-l-thyronine (rT3), 3,5-diiodo-l-thyronine (3,5-T2) and 3,3'-diiodo-l-thyronine (3,3'-T2) in zebrafish larvae was achieved by ultra-performance liquid chromatography-tandem mass spectrometry in positive ion mode. Solid-phase extraction with SampliQ cartridges and derivatization with 3 m hydrochloric acid in n-butanol reduced matrix effects. Derivatized compounds were separated on an Acquity UPLC BEH C 18 column with mobile phases consisting of 0.1% acetic acid in deionized water and 0.1% acetic acid in methanol. The limits of detection ranged from 0.5 to 0.6 pg injected on column. The method was validated by evaluating recovery (77.1-117.2%), accuracy (87.3-123.9%) and precision (0.5-12.4%) using diluted homogenized zebrafish embryos spiked with all target compounds. This method was then applied to zebrafish larvae collected after 114 h of exposure to polychlorinated biphenyls (PCBs), including PCB 28, PCB 66 and PCB 95, or the technical mixture Aroclor 1254. Exposure to PCB 28 and PCB 95 increased the T4:T3 ratio and decreased the T3:rT3 ratio, demonstrating that this method can effectively detect PCB-induced alterations in THs. Copyright © 2018 John Wiley & Sons, Ltd.

Unmasking tandem site interaction in human acetylcholinesterase. Substrate activation with a cationic acetanilide substrate.

PubMed

Johnson, Joseph L; Cusack, Bernadette; Davies, Matthew P; Fauq, Abdul; Rosenberry, Terrone L

2003-05-13

Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge, and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acyl enzyme intermediate is produced. A conformational interaction between the A- and P-sites has recently been found to modulate ligand affinities. We now demonstrate that this interaction is of functional importance by showing that the acetylation rate constant of a substrate bound to the A-site is increased by a factor a when a second molecule of substrate binds to the P-site. This demonstration became feasible through the introduction of a new acetanilide substrate analogue of acetylcholine, 3-(acetamido)-N,N,N-trimethylanilinium (ATMA), for which a = 4. This substrate has a low acetylation rate constant and equilibrates with the catalytic site, allowing a tractable algebraic solution to the rate equation for substrate hydrolysis. ATMA affinities for the A- and P-sites deduced from the kinetic analysis were confirmed by fluorescence titration with thioflavin T as a reporter ligand. Values of a >1 give rise to a hydrolysis profile called substrate activation, and the AChE site-specific mutant W86F, and to a lesser extent wild-type human AChE itself, showed substrate activation with acetylthiocholine as the substrate. Substrate activation was incorporated into a previous catalytic scheme for AChE in which a bound P-site ligand can also block product dissociation from the A-site, and two additional features of the AChE catalytic pathway were revealed. First, the ability of a bound P-site ligand to increase the substrate acetylation rate constant varied with the structure of the ligand: thioflavin T accelerated ATMA acetylation by a factor a(2) of 1.3, while propidium failed to accelerate. Second, catalytic rate

DEVELOPMENTAL AND WITHDRAWAL EFFECTS OF ADOLESCENT AAS EXPOSURE ON THE GLUTAMATERGIC SYSTEM IN HAMSTERS

PubMed Central

Carrillo, Maria; Ricci, Lesley A.; Melloni, Richard H.

2011-01-01

In the Syrian hamster (Mesocricetus auratus) glutamate activity has been implicated in the modulation of adolescent anabolic-androgenic steroid (AAS)-induced aggression. The current study investigated the time course of adolescent AAS-induced neurodevelopmental and withdrawal effects on the glutamatergic system and examined whether these changes paralleled those of adolescent AAS-induced aggression. Glutamate activity in brain areas comprising the aggression circuit in hamsters and aggression were examined following 1, 2, 3 and 4 weeks of AAS treatment or 1, 2, 3 and 4 weeks following the cessation of AAS exposure. In these studies glutamate activity was examined using vesicular glutamate transporter 2 (VGLUT2). The onset of aggression was observed following 2 weeks exposure to AAS and continued to increase showing maximal aggression levels after 4 weeks of AAS treatment. This aggressive phenotype was detected after 2 weeks of withdrawal from AAS. The time-course of AAS-induced changes in latero anterior hypothalamus (LAH)-VGLUT2 closely paralleled increases in aggression. Increases in LAH-VGLUT2 were first detected in animals exposed to AAS for 2 weeks and were maintained up to 3 weeks following the cessation of AAS treatment. AAS treatment also produced developmental and long-term alterations in VGLUT2 expression within other aggression areas. However, AAS-induced changes in glutamate activity within these regions did not coincide with changes in aggression. Together, these data indicate that adolescent AAS treatment leads to alterations in the glutamatergic system in brain areas implicated in aggression control, yet only alterations in LAH-glutamate parallel the time course of AAS-induced changes in the aggressive phenotype. PMID:21500881

Detection of AA76, a Common Form of Amyloid A Protein, as a Way of Diagnosing AA Amyloidosis.

PubMed

Sato, Junji; Okuda, Yasuaki; Kuroda, Takeshi; Yamada, Toshiyuki

2016-01-01

Reactive amyloid deposits consist of amyloid A (AA) proteins, the degradation products of serum amyloid A (SAA). Since the most common species of AA is the amino terminal portion produced by cleavage between residues 76 and 77 of SAA (AA76), the presence of AA76 in tissues could be a consequence of AA amyloid deposition. This study assessed the diagnostic significance of the detection of AA76 for AA amyloidosis using two different approaches. Biopsy specimens (n=130 from 54 subjects) from gastroduodenal mucosa or abdominal fat (n=9 from 9 subjects) of patients who had already been diagnosed with or were suspected of having AA amyloidosis were used. Fixed mucosal sections were subjected to immunohistochemistry using a newly developed antibody recognizing the carboxyl terminal end of AA76 (anti-AA76). The non-fixed materials from gastroduodenal mucosa or abdominal fat were subjected to immunoblotting for detection of the size of AA76. Among the gastroduodenal specimens (n=115) from already diagnosed patients, the positive rates of Congo red staining, immunohistochemistry using anti-AA76, and immunoblotting were 68.4%, 73.0%, and 92.2%, respectively. The anti-AA76 did not stain the supposed SAA in the blood or leakage, which was stained by anti-SAA antibody. AA76 was not detected either by immunohistochemistry or by immunoblot in the materials from patients in whom AA amyloidosis had been ruled out. In the abdominal fat, the immunoblot detected AA76 in 8 materials from 8 already diagnosed patients and did not in 1 patient whose gastroduodenal mucosa was negative. In conclusion, the detection of AA76 may alter the ability to diagnose AA amyloidosis. In immunohistochemistry for fixed specimens, the new anti-AA76 antibody can improve the specificity. Immunoblot for non-fixed materials, which can considerably improve the sensitivity, should be beneficial for small materials like abdominal fat. © 2016 by the Association of Clinical Scientists, Inc.

Fatigue crack growth in fiber-metal laminates

NASA Astrophysics Data System (ADS)

Ma, YuE; Xia, ZhongChun; Xiong, XiaoFeng

2014-01-01

Fiber-metal laminates (FMLs) consist of three layers of aluminum alloy 2024-T3 and two layers of glass/epoxy prepreg, and it (it means FMLs) is laminated by Al alloy and fiber alternatively. Fatigue crack growth rates in notched fiber-metal laminates under constant amplitude fatigue loading were studied experimentally and numerically and were compared with them in monolithic 2024-T3 Al alloy plates. It is shown that the fatigue life of FMLs is about 17 times longer than monolithic 2024-T3 Al alloy plate; and crack growth rates in FMLs panels remain constant mostly even when the crack is long, unlike in the monolithic 2024-T3 Al alloy plates. The formula to calculate bridge stress profiles of FMLs was derived based on the fracture theory. A program by Matlab was developed to calculate the distribution of bridge stress in FMLs, and then fatigue growth lives were obtained. Finite element models of FMLs were built and meshed finely to analyze the stress distributions. Both results were compared with the experimental results. They agree well with each other.

A biomarker-responsive T2ex MRI contrast agent.

PubMed

Daryaei, Iman; Randtke, Edward A; Pagel, Mark D

2017-04-01

This study investigated a fundamentally new type of responsive MRI contrast agent for molecular imaging that alters T 2 exchange (T 2ex ) properties after interacting with a molecular biomarker. The contrast agent Tm-DO3A-oAA was treated with nitric oxide (NO) and O 2 . The R 1 and R 2 relaxation rates of the reactant and product were measured with respect to concentration, temperature, and pH. Chemical exchange saturation transfer (CEST) spectra of the reactant and product were acquired using a 7 Tesla (T) MRI scanner and analyzed to estimate the chemical exchange rates and r 2ex relaxivities. The reaction of Tm-DO3A-oAA with NO and O 2 caused a 6.4-fold increase in the r 2 relaxivity of the agent, whereas r 1 relaxivity remained unchanged, which demonstrated that Tm-DO3A-oAA is a responsive T 2ex agent. The effects of pH and temperature on the r 2 relaxivities of the reactant and product supported the conclusion that the product's benzimidazole ligand caused the agent to have a fast chemical exchange rate relative to the slow exchange rate of the reactant's ortho-aminoanilide ligand. T 2ex MRI contrast agents are a new type of responsive agent that have good detection sensitivity and specificity for detecting a biomarker, which can serve as a new tool for molecular imaging. Magn Reson Med 77:1665-1670, 2017. © 2016 International Society for Magnetic Resonance in Medicine. © 2016 International Society for Magnetic Resonance in Medicine.

Plasmodium falciparum mitochondria import tRNAs along with an active phenylalanyl-tRNA synthetase.

PubMed

Sharma, Arvind; Sharma, Amit

2015-02-01

The Plasmodium falciparum protein translation enzymes aminoacyl-tRNA synthetases (aaRSs) are an emergent family of drug targets. The aaRS ensemble catalyses transfer of amino acids to cognate tRNAs, thus providing charged tRNAs for ribosomal consumption. P. falciparum proteome expression relies on a total of 36 aaRSs for the three translationally independent compartments of cytoplasm, apicoplast and mitochondria. In the present study, we show that, of this set of 36, a single genomic copy of mitochondrial phenylalanyl-tRNA synthetase (mFRS) is targeted to the parasite mitochondria, and that the mFRS gene is exclusive to malaria parasites within the apicomplexan phyla. Our protein cellular localization studies based on immunofluorescence data show that, along with mFRS, P. falciparum harbours two more phenylalanyl-tRNA synthetase (FRS) assemblies that are localized to its apicoplast and cytoplasm. The 'extra' mFRS is found in mitochondria of all asexual blood stage parasites and is competent in aminoacylation. We show further that the parasite mitochondria import tRNAs from the cytoplasmic tRNA pool. Hence drug targeting of FRSs presents a unique opportunity to potentially stall protein production in all three parasite translational compartments.

PDGF-AA-induced filamentous mitochondria benefit dermal papilla cells in cellular migration.

PubMed

Mifude, C; Kaseda, K

2015-06-01

Human dermal papilla cells (HDPCs) play essential roles in hair follicular morphogenesis and postnatal hair growth cycles. Previous reports demonstrated that platelet-derived growth factor-AA (PDGF-AA) enhanced the formation of dermal condensates in hair follicular development. Additionally, PDGF-AA induces/maintains the anagen phase of the hair cycle. It is likely that mitochondrial morphology and functions are tightly coupled with maintenance of these energy-demanding activities. However, little is known about the mitochondrial regulation in HDPCs. Thus, we investigated the PDGF-involved mitochondrial regulation in HDPCs. The mitochondrial morphologies of HDPCs were examined in the presence or absence of PDGF-AA under a fluorescent microscope. ATP production and cellular motility were investigated. The relationship between mitochondrial morphology and the cellular functions was discussed. We observed that primary HDPCs contained mitochondria with filamentous and/or rounded morphologies. Both types of mitochondria showed similar membrane potentials. Interestingly, in the presence of PDGF-AA, but not PDGF-BB, the balance between the two morphologies shifted towards the filamentous form. Concomitantly, both mitochondrial enzymatic activity and total cellular ATP level were augmented by PDGF-AA. These two parameters were closely correlated, suggesting the mitochondrial involvement in the PDGF-augmented ATP production. Moreover, PDGF-AA accelerated the migration of HDPCs in a gap-filling assay, but did not change the rate of cellular proliferation. Notably, filamentous mitochondria dominated migrating HDPCs. PDGF-AA benefits HDPCs in the process of migration, by increasing the number of filamentous mitochondria. © 2014 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

Gamma-tocotrienol attenuates the aberrant lipid mediator production in NLRP3 inflammasome-stimulated macrophages.

PubMed

Kim, Yongeun; Gromovsky, Anthony D; Brown, J Mark; Chung, Soonkyu

2018-06-04

The activation of NLRP3 inflammasome in innate immune cells is associated with enhanced production of pro-inflammatory lipid mediator eicosanoids that play a crucial role in propagating inflammation. Gamma-tocotrienol (γT3) is an unsaturated vitamin E that has been demonstrated to attenuate NLRP3-inflammasome. However, the role of γT3 in regulating eicosanoid formation is unknown. We hypothesized that γT3 abolishes the eicosanoid production by modulating the macrophage lipidome. LPS-primed bone marrow-derived macrophages (BMDM) were stimulated with saturated fatty acids (SFA) along with γT3, and the effects of γT3 in modulating macrophage lipidome were quantified by using mass spectrometry based-shotgun lipidomic approaches. The SFA-mediated inflammasome activation induced robust changes in lipid species of glycerolipids (GL), glycerophospholipids (GPL), and sphingolipids in BMDM, which were distinctly different in the γT3-treated BMDM. The γT3 treatment caused substantial decreases of lysophospholipids (LysoPL), diacylglycerol (DAG), and free arachidonic acid (AA, C20:4), indicating that γT3 limits the availability of AA, the precursor for eicosanoids. This was confirmed by the pulse-chase experiment using [ 3 H]-AA, and by diminished prostaglandin E 2 (PGE 2 ) secretion by ELISA. Concurrently, γT3 inhibited LPS-induced cyclooxygenases 2 (COX2) induction, further suppressing prostaglandin synthesis. In addition, γT3 attenuated ceramide synthesis by transcriptional downregulation of key enzymes for de novo synthesis. The altered lipid metabolism during inflammation is linked to reduced ATP production, which was partly rescued by γT3. Taken together, our work revealed that γT3 induces distinct modification of the macrophage lipidome to reduce AA release and corresponding lipid mediator synthesis, leading to attenuated cellular lipotoxicity. Copyright © 2018 Elsevier Inc. All rights reserved.

Association of aplastic anemia and FoxP3 gene polymorphisms in Koreans.

PubMed

In, Ji Won; Lee, Nuri; Roh, Eun Youn; Shin, Sue; Park, Kyoung Un; Song, Eun Young

2017-04-01

Aplastic anemia (AA) is characterized by pancytopenia and bone marrow failure, and most acquired AA is an immune-mediated disorder. Regulatory T cells (T regs ) suppressing autoreactive T cells were decreased in AA patients. FoxP3 is a major regulator for the development and function of T regs . Polymorphism in FoxP3 was shown to be associated with various autoimmune diseases, however, has not yet been studied in AA. In this study, we examined the association between FoxP3 polymorphisms and AA in Korean patients. The study population consisted of 94 patients diagnosed by bone marrow examination in Seoul National University Hospital (SNUH) during 1997-2012 and 195 healthy controls. FoxP3 polymorphisms (rs5902434 del/ATT, rs3761548 C/A, rs3761549 C/T, rs2232365 A/G) were analyzed by PCR-sequencing method. We analyzed differences of genotype and allele frequencies between patients and controls. We also compared differences of genotype and allele frequencies between responder and non-responder in patients treated with immunosuppressive therapy (IST). For the statistical analysis, the chi-square test and Fisher's exact test were used and P < 0.05 was regarded as statistically significant. There was no significant difference in the genotype frequencies of FoxP3 polymorphisms between patients and controls. With regards to the allele frequencies, rs3761548 C allele was significantly higher in AA patients than in controls (87.4% vs. 79.7%, P = 0.047). In patients treated with IST, rs3761549 C allele was significantly higher in non-responder patients than in responders (89.6% vs. 66.7%, P = 0.036) and female rs3761549 C/C genotype carriers were associated with greater risk for non-response to IST (84.2% vs. 16.7%, P = 0.006). Polymorphisms in rs3761548 and rs3761549 of FoxP3 in our population were associated with disease susceptibility and response for IST, respectively. This study suggests an association between FoxP3 polymorphisms and AA in Korean patients

A near-infrared luminescent Mn2+-doped NaYF4:Yb,Tm/Fe3+ upconversion nanoparticles redox reaction system for the detection of GSH/Cys/AA.

PubMed

Zhang, Liping; Ling, Bo; Wang, Lun; Chen, Hongqi

2017-09-01

An upconversion luminescence method was developed for the determination of glutathione (GSH), L-cysteine (Cys) or L-ascorbic acid (AA) based on redox reaction. We synthesized poly(acrylic acid) (PAA)-modified Mn 2+ -doped NaYF 4 :Yb,Tm upconversion nanoparticles (UCNPs), and the luminescence of these UCNPs was effectively quenched due to their carboxyl groups coordinating with Fe 3+ to form a UCNPs/Fe 3+ system. GSH, Cys or AA reduced Fe 3+ to Fe 2+ , which induced the luminescence recovery of the UCNPs. Under the optimized conditions, wide linear concentration ranges from 0.25-300μM for GSH, 0.5-875μM for Cys and 0.5-350μM for AA were found, and the detection limits (3S/K) were 0.2μM, 0.5μM and 0.2μM, respectively. Thus, the UCNPs/Fe 3+ system was successfully applied for sensing GSH, Cys or AA. Copyright © 2017 Elsevier B.V. All rights reserved.

tRNAHis guanylyltransferase catalyzes a 3'-5' polymerization reaction that is distinct from G-1 addition.

PubMed

Jackman, Jane E; Phizicky, Eric M

2006-06-06

Yeast tRNA(His) guanylyltransferase, Thg1, is an essential protein that adds a single guanine to the 5' end (G(-1)) of tRNA(His). This G(-1) residue is required for aminoacylation of tRNA(His) by histidyl-tRNA synthetase, both in vitro and in vivo. The guanine nucleotide addition reaction catalyzed by Thg1 extends the polynucleotide chain in the reverse (3'-5') direction of other known polymerases, albeit by one nucleotide. Here, we show that alteration of the 3' end of the Thg1 substrate tRNA(His) unleashes an unexpected reverse polymerase activity of wild-type Thg1, resulting in the 3'-5' addition of multiple nucleotides to the tRNA, with efficiency comparable to the G(-1) addition reaction. The addition of G(-1) forms a mismatched G.A base pair at the 5' end of tRNA(His), and, with monophosphorylated tRNA substrates, it is absolutely specific for tRNA(His). By contrast, reverse polymerization forms multiple G.C or C.G base pairs, and, with preactivated tRNA species, it can initiate at positions other than -1 and is not specific for tRNA(His). Thus, wild-type Thg1 catalyzes a templated polymerization reaction acting in the reverse direction of that of canonical DNA and RNA polymerases. Surprisingly, Thg1 can also readily use dNTPs for nucleotide addition. These results suggest that 3'-5' polymerization represents either an uncharacterized role for Thg1 in RNA or DNA repair or metabolism, or it may be a remnant of an earlier catalytic strategy used in nature.

AA amyloidosis complicating the hereditary periodic fever syndromes.

PubMed

Lane, Thirusha; Loeffler, Jutta M; Rowczenio, Dorota M; Gilbertson, Janet A; Bybee, Alison; Russell, Tonia L; Gillmore, Julian D; Wechalekar, Ashutosh D; Hawkins, Philip N; Lachmann, Helen J

2013-04-01

AA amyloidosis is a life-threatening complication of the hereditary periodic fever syndromes (HPFS), which are otherwise often compatible with normal life expectancy. This study was undertaken to determine the characteristics, presentation, natural history, and response to treatment in 46 patients who had been referred for evaluation at the UK National Amyloidosis Centre. Disease activity was monitored by serial measurement of serum amyloid A. Renal function was assessed by measurement of serum creatinine and albumin levels, the estimated glomerular filtration rate, and proteinuria from 24-hour urine collections. The amyloid load was measured by serum amyloid P scintigraphy. Twenty-four patients had familial Mediterranean fever, 12 patients had tumor necrosis factor receptor-associated periodic syndrome, 6 patients had cryopyrin-associated periodic syndromes, and 4 patients had mevalonate kinase deficiency. The median age at onset of HPFS was 5 years; median age at presentation with AA amyloidosis was 38 years. Diagnosis of an HPFS had not been considered prior to presentation with AA amyloidosis in 23 patients (50%). Eleven patients (24%) had end-stage renal failure (ESRF) at presentation; of these, 3 had received transplants prior to referral. A further 13 patients developed ESRF over the followup period, with 10 undergoing renal transplantation. The median time to progression to ESRF from onset of AA amyloidosis was 3.3 years (interquartile range [IQR] 2-8), with a median time to transplant of 4 years (IQR 3-6). Eleven patients (24%) died. The median survival in the entire cohort was 19 years from diagnosis of AA amyloidosis. Of the 37 patients who were treated successfully, or in whom at least partial suppression of the underlying HPFS was achieved, 17 (46%) showed amyloid regression, 14 (38%) showed a stable amyloid load, and 2 (5%) showed increased amyloid deposition over the followup period. AA amyloidosis remains a challenging and serious late complication

Insulin receptor substrate-2 gene variants in subjects with metabolic syndrome: association with plasma monounsaturated and n-3 polyunsaturated fatty acid levels and insulin resistance.

PubMed

Perez-Martinez, Pablo; Delgado-Lista, Javier; Garcia-Rios, Antonio; Tierney, Audrey C; Gulseth, Hanne L; Williams, Christine M; Karlström, Brita; Kieć-Wilk, Beata; Blaak, Ellen E; Helal, Olfa; Saris, Wim H M; Defoort, Catherine; Drevon, Christian A; Lovegrove, Julie A; Dembinska-Kieć, Aldona; Riserus, Ulf; Roche, Helen M; Lopez-Miranda, Jose

2012-02-01

Several insulin receptor substrate-2 (IRS-2) polymorphisms have been studied in relation to insulin resistance and type 2 diabetes. To examine whether the genetic variability at the IRS-2 gene locus was associated with the degree of insulin resistance and plasma fatty acid levels in metabolic syndrome (MetS) subjects. Insulin sensitivity, insulin secretion, glucose effectiveness, plasma fatty acid composition and three IRS-2 tag-single nucleotide polymorphisms (SNPs) were determined in 452 MetS subjects. Among subjects with the lowest level of monounsaturated (MUFA) (below the median), the rs2289046 A/A genotype was associated with lower glucose effectiveness (p<0.038), higher fasting insulin concentrations (p<0.028) and higher HOMA IR (p<0.038) as compared to subjects carrying the minor G-allele (A/G and G/G). In contrast, among subjects with the highest level of MUFA (above the median), the A/A genotype was associated with lower fasting insulin concentrations and HOMA-IR, whereas individuals carrying the G allele and with the highest level of ω-3 polyunsaturated fatty acids (above the median) showed lower fasting insulin (p<0.01) and HOMA-IR (p<0.02) as compared with A/A subjects. The rs2289046 polymorphism at the IRS2 gene locus may influence insulin sensitivity by interacting with certain plasma fatty acids in MetS subjects. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Structure of the Human FANCL RING-Ube2T Complex Reveals Determinants of Cognate E3-E2 Selection

PubMed Central

Hodson, Charlotte; Purkiss, Andrew; Miles, Jennifer Anne; Walden, Helen

2014-01-01

Summary The combination of an E2 ubiquitin-conjugating enzyme with an E3 ubiquitin-ligase is essential for ubiquitin modification of a substrate. Moreover, the pairing dictates both the substrate choice and the modification type. The molecular details of generic E3-E2 interactions are well established. Nevertheless, the determinants of selective, specific E3-E2 recognition are not understood. There are ∼40 E2s and ∼600 E3s giving rise to a possible ∼24,000 E3-E2 pairs. Using the Fanconi Anemia pathway exclusive E3-E2 pair, FANCL-Ube2T, we report the atomic structure of the FANCL RING-Ube2T complex, revealing a specific and extensive network of additional electrostatic and hydrophobic interactions. Furthermore, we show that these specific interactions are required for selection of Ube2T over other E2s by FANCL. PMID:24389026

Some observations on loss of static strength due to fatigue cracks

NASA Technical Reports Server (NTRS)

Illg, Walter; Hardrath, Herbert F

1955-01-01

Static tensile tests were performed on simple notched specimens containing fatigue cracks. Four types of aluminum alloys were investigated: 2024-T3(formerly 24S-T3) and 7075-T6(formerly 75S-T6) in sheet form, and 2024-T4(formerly 24S-T4) and 7075-T6(formerly 75S-T6) in extruded form. The cracked specimens were tested statically under four conditions: unmodified and with reduced eccentricity of loading by three methods. Results of static tests on C-46 wings containing fatigue cracks are also reported.

Controlled Release from Core-Shell Nanoporous Silica Particles for Corrosion Inhibition of Aluminum Alloys

DOE PAGES

Jiang, Xingmao; Jiang, Ying-Bing; Liu, Nanguo; ...

2011-01-01

Ceriumore » m (Ce) corrosion inhibitors were encapsulated into hexagonally ordered nanoporous silica particles via single-step aerosol-assisted self-assembly. The core/shell structured particles are effective for corrosion inhibition of aluminum alloy AA2024-T3. Numerical simulation proved that the core-shell nanostructure delays the release process. The effective diffusion coefficient elucidated from release data for monodisperse particles in water was 1.0 × 10 − 14  m 2 s for Ce 3+ compared to 2.5 × 10 − 13  m 2 s for NaCl. The pore size, pore surface chemistry, and the inhibitor solubility are crucial factors for the application. Microporous hydrophobic particles encapsulating a less soluble corrosion inhibitor are desirable for long-term corrosion inhibition.« less

Functional and computational analysis of amino acid patterns predictive of type III secretion system substrates in Pseudomonas syringae

USDA-ARS?s Scientific Manuscript database

Bacterial type III secretion systems (T3SSs) deliver proteins called effectors into eukaryotic cells. Although N-terminal amino acid sequences are required for translocation, the mechanism of substrate recognition by the T3SS is unknown. Almost all actively deployed T3SS substrates in the plant path...

Constructing honeycomb micropatterns on nonplanar substrates with high glass transition temperature polymers.

PubMed

Ding, Jianyun; Gong, Jianliang; Bai, Hua; Li, Lei; Zhong, Yawen; Ma, Zhi; Svrcek, Vladimir

2012-08-15

In Qiao's previous report, only star polymers with T(g) (glass transition temperature) below 48°C were found forming homogeneous honeycomb coatings on the nonplanar substrates. The polymers with high T(g) are believed not able to duplicate nonplanar substrate due to their brittleness. This article presents a comprehensive study on the construction of macroporous polymeric films on various nonplanar substrates with static breath figure (BF) technique, using linear polymers with high T(g). Two kinds of linear polymers with high T(g), polystyrene-b-poly(acrylic acid) and polystyrene without polar end groups, are employed to prepare 3-dimensional macroporous films on different nonplanar substrates. Scanning electronic microscopy views on the side wall in addition to views in-plane prove that polymer films with BF array perfectly replicated the surface features of these substrates. The formation processes of macropores on these substrates are analyzed in detail, and it demonstrates that neither molecular topography nor T(g) of polymers is the critical factor contouring nonplanar substrate. A new hypothesis involving polymer plasticization and conformation during the solvent evaporation is formulated. Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.

USAF Damage Tolerant Design Handbook: Guidelines for the Analysis and Design of Damage Tolerant Aircraft Structures. Revision B

DTIC Science & Technology

1984-05-01

Effect of Block Programming and Block Size on Crack Growth Life (All Histories Have Same Cycle Content) Alloy : 2024 -T3 Aluminum (Reference 27...1 I 6 o T 1 >0 100 Crack Growth Life for l-in. Crack Growth 1000 Figure 5.1.10, Effect of Humidity on Fatigue Crack Propagation...Growth Life (All Histories Have Same Cycle Content) Alloy : 2024 -T3 Aluminum (Reference 27). 5.2.6 Yield Zone Due to Overload (r T), Current Crack

Functional Connectivity Substrates for tDCS Response in Minimally Conscious State Patients

PubMed Central

Cavaliere, Carlo; Aiello, Marco; Di Perri, Carol; Amico, Enrico; Martial, Charlotte; Thibaut, Aurore; Laureys, Steven; Soddu, Andrea

2016-01-01

Transcranial direct current stimulation (tDCS) is a non-invasive technique recently employed in disorders of consciousness, and determining a transitory recovery of signs of consciousness in almost half of minimally conscious state (MCS) patients. Although the rising evidences about its possible role in the treatment of many neurological and psychiatric conditions exist, no evidences exist about brain functional connectivity substrates underlying tDCS response. We retrospectively evaluated resting state functional Magnetic Resonance Imaging (fMRI) of 16 sub-acute and chronic MCS patients (6 tDCS responders) who successively received a single left dorsolateral prefrontal cortex (DLPFC) tDCS in a double-blind randomized cross-over trial. A seed-based approach for regions of left extrinsic control network (ECN) and default-mode network (DMN) was performed. tDCS responders showed an increased left intra-network connectivity for regions co-activated with left DLPFC, and significantly with left inferior frontal gyrus. Non-responders (NR) MCS patients showed an increased connectivity between left DLPFC and midline cortical structures, including anterior cingulate cortex and precuneus. Our findings suggest that a prior high connectivity with regions belonging to ECN can facilitate transitory recovery of consciousness in a subgroup of MCS patients that underwent tDCS treatment. Therefore, resting state-fMRI could be very valuable in detecting the neuronal conditions necessary for tDCS to improve behavior in MCS. PMID:27857682

Functional Connectivity Substrates for tDCS Response in Minimally Conscious State Patients.

PubMed

Cavaliere, Carlo; Aiello, Marco; Di Perri, Carol; Amico, Enrico; Martial, Charlotte; Thibaut, Aurore; Laureys, Steven; Soddu, Andrea

2016-01-01

Transcranial direct current stimulation (tDCS) is a non-invasive technique recently employed in disorders of consciousness, and determining a transitory recovery of signs of consciousness in almost half of minimally conscious state (MCS) patients. Although the rising evidences about its possible role in the treatment of many neurological and psychiatric conditions exist, no evidences exist about brain functional connectivity substrates underlying tDCS response. We retrospectively evaluated resting state functional Magnetic Resonance Imaging (fMRI) of 16 sub-acute and chronic MCS patients (6 tDCS responders) who successively received a single left dorsolateral prefrontal cortex (DLPFC) tDCS in a double-blind randomized cross-over trial. A seed-based approach for regions of left extrinsic control network (ECN) and default-mode network (DMN) was performed. tDCS responders showed an increased left intra-network connectivity for regions co-activated with left DLPFC, and significantly with left inferior frontal gyrus. Non-responders (NR) MCS patients showed an increased connectivity between left DLPFC and midline cortical structures, including anterior cingulate cortex and precuneus. Our findings suggest that a prior high connectivity with regions belonging to ECN can facilitate transitory recovery of consciousness in a subgroup of MCS patients that underwent tDCS treatment. Therefore, resting state-fMRI could be very valuable in detecting the neuronal conditions necessary for tDCS to improve behavior in MCS.

Laboratory evaluation of transgenic Populus davidiana×Populus bolleana expressing Cry1Ac + SCK, Cry1Ah3, and Cry9Aa3 genes against gypsy moth and fall webworm.

PubMed

Ding, Liping; Chen, Yajuan; Wei, Xiaoli; Ni, Mi; Zhang, Jiewei; Wang, Hongzhi; Zhu, Zhen; Wei, Jianhua

2017-01-01

Transgenic poplar lines 'Shanxin' (Populus davidiana×Populus bolleana) were generated via Agrobacterium-mediated transformation. The transgenic lines carried the expression cassettes of Cry1Ac + SCK, Cry1Ah3, and Cry9Aa3, respectively. The expression levels of the exogenous insect resistance genes in the transgenic lines were determined by Q-PCR and Western blot. Leaves of the transgenic lines were used for insect feeding bioassays on first instar larvae of the gypsy moth (Lymantria dispar) and fall webworm (Hyphantria cunea). At 5 d of feeding, the mean mortalities of larvae feeding on Cry1Ac + SCK and Cry1Ah3 transgenic poplars leaves were 97% and 91%, while mortality on Cry9Aa3 transgenic lines was about 49%. All gypsy moth and fall webworm larvae were killed in 7-9 days after feeding on leaves from Cry1Ac + SCK or Cry1Ah3 transgenic poplars, while all the fall webworm larvae were killed in 11 days and about 80% of gypsy moth larvae were dead in 14 days after feeding on those from Cry9Aa3 transgenic lines. It was concluded that the transgenic lines of Cry1Ac + SCK and Cry1Ah3 were highly toxic to larvae of both insect species while lines with Cry9Aa3 had lower toxicity,and H. cunea larvae are more sensitive to the insecticidal proteins compared to L. dispar. Transgenic poplar lines toxic to L. dispar and H. cunea could be used to provide Lepidoptera pest resistance to selected strains of poplar trees.

Trace metal assay of U(3)O(8) powder by electrothermal AAS.

PubMed

Page, A G; Godbole, S V; Kulkarni, M J; Porwal, N K; Shelar, S S; Joshi, B D

1983-10-01

Methods have been developed for the direct determination of Ag, Ca, K., Li, Mg, Na, Pb, Sn and Zn in U(3)O(8) powder samples by electrothermal AAS. Nanogram and lower amounts of these elements have been determined with a relative standard deviation of 6-16% in mg amounts of sample (either alone or mixed with an equal weight of graphite). The results for NBL reference samples were in reasonable agreement with the certified values. X-Ray diffraction studies on the residues left from the graphite mixtures after the atomization cycle, confirmed the formation of uranium carbide (UC(2)).

Role of Transmembrane Domain 8 in Substrate Selectivity and Translocation of SteT, a Member of the l-Amino Acid Transporter (LAT) Family*

PubMed Central

Bartoccioni, Paola; del Rio, César; Ratera, Merce; Kowalczyk, Lukasz; Baldwin, Jocelyn M.; Zorzano, Antonio; Quick, Matthias; Baldwin, Stephen A.; Vázquez-Ibar, José Luis; Palacín, Manuel

2010-01-01

System l-amino acid transporters (LAT) belong to the amino acid, polyamine, and organic cation superfamily of transporters and include the light subunits of heteromeric amino acid transporters and prokaryotic homologues. Cysteine reactivity of SteT (serine/threonine antiporter) has been used here to study the substrate-binding site of LAT transporters. Residue Cys-291, in transmembrane domain 8 (TM8), is inactivated by thiol reagents in a substrate protectable manner. Surprisingly, DTT activated the transporter by reducing residue Cys-291. Cysteine-scanning mutagenesis of TM8 showed DTT activation in the single-cysteine mutants S287C, G294C, and S298C, lining the same α-helical face. S-Thiolation in Escherichia coli cells resulted in complete inactivation of the single-cysteine mutant G294C. l-Serine blocked DTT activation with an EC50 similar to the apparent KM of this mutant. Thus, S-thiolation abolished substrate translocation but not substrate binding. Mutation of Lys-295, to Cys (K295C) broadened the profile of inhibitors and the spectrum of substrates with the exception of imino acids. A structural model of SteT based on the structural homologue AdiC (arginine/agmatine antiporter) positions residues Cys-291 and Lys-295 in the putative substrate binding pocket. All this suggests that Lys-295 is a main determinant in the recognition of the side chain of SteT substrates. In contrast, Gly-294 is not facing the surface, suggesting conformational changes involving TM8 during the transport cycle. Our results suggest that TM8 sculpts the substrate-binding site and undergoes conformational changes during the transport cycle of SteT. PMID:20610400

[Risk control of traditional Chinese medicines containing aristolochis acids (AAs) based on influencing factors of content of AAs].

PubMed

Tian, Jing-Zhuo; Liang, Ai-Hua; Liu, Jing; Zhang, Bo-Li

2017-12-01

Aristolochic acids (AAs) widely exist in such plants as Aristolochia and Asarum. The renal toxicity of AAs as well as its carcinogenicity to urinary system have been widely known. In 2003 and 2004, China prohibited the use of Aristolochiae Radix, Aristolochiae Manshuriensis Caulis and Aristolochiae Fangchi Radix, and required administering other AAs-containing medicines in accordance with the regulations for prescription drugs. In this paper, we retrieved literatures on the content determination of AAs in recent 10 years in China. It suggested that the AAs content is lower in Asarum herb, especially in its roots and rhizomes, and most of which do not show detectable amount of AA-I. Some of traditional Chinese medicines show fairly small amount of detectable AA-I. The AAs content in Aristolochia herb (including Fructus Aristolochiae, kaempfer dutchmanspipe root) is relatively high; however, there are fewer literatures for studying the content determination of AAs in Chinese patent medicines. There were many factors affecting AAs content, including the parts used, origins, processing methods, extraction process. It suggested that we should pay attention to the toxicity of Chinese medicines containing AAs and use these decoction pieces and traditional Chinese medicines cautiously. In addition, basic studies for the origins, processing methods and extraction process of Chinese patent medicines containing AAs, as well as supervision and detection of AAs content in traditional Chinese medicinal materials, decoction pieces and Chinese patent medicines shall be strengthened for reducing medication risk and guaranteeing clinical medication safety. Copyright© by the Chinese Pharmaceutical Association.

Characterization of monoclonal antibodies against hepatitis C virus nonstructural protein 3: different antigenic determinants from human B cells.

PubMed

Ou-Yang, P; Chiang, B L; Hwang, L H; Chen, Y G; Yang, P M; Chi, W K; Chen, P J; Chen, D S

1999-04-01

The nonstructural (NS3) region protein of hepatitis C virus (HCV) possesses major B-cell epitopes that induce antibodies after infection. To elucidate further the characteristics of these B cells and their role in the immune regulation of HCV infection, T9 (portion of NS3 region, amino acids [a.a.] 1188-1493)-specific monoclonal antibodies were derived and mapped for B-cell antigenic determinants with recombinant proteins. A total of 10 T9-specific hybridomas were generated and tested for B-cell antigenic determinants. To analyze the B-cell antigenic determinants, eight recombinant proteins including NS3-e (a.a. 1175-1334), NS3-a' (a.a. 1175-1250), NS3-a (a.a. 1251-1334), NS3-b (a.a. 1323-1412), NS3-c (a.a. 1407-1499), NS3-a/b (a.a. 1251-1412), NS3-bc (a.a. 1323-1499), and NS3-abc (a.a. 1251-1499) encoded by NS3-region internal clones were expressed and tested for immunoblotting. The data suggested IgG hybridomas recognized NS3-a, NS3-a', or NS3-b protein by immunoblotting. By contrast, the NS3-e protein bears the major antigenic determinant recognized by human sera. Half of the hybridomas were found to react with protein NS3-a', which is not a major B-cell antigenic determinant in humans. These data suggested that conformational epitopes in vivo may be important for B-cell recognition.

Active site nanospace of aminoacyl tRNA synthetase: difference between the class I and class II synthetases.

PubMed

Dutta, Saheb; Choudhury, Kaberi; Banik, Sindrila Dutta; Nandi, Nilashis

2014-03-01

The present work is aimed at understanding the origin of the difference in the molecular organization of the active site nanospaces of the class I and class II aminoacyl tRNA synthetases (aaRSs) which are tunnel-like structures. The active site encloses the cognate amino acid (AA) and the adenosine triphosphate (ATP) to carry out aminoacylation reaction. Comparison of the structures of the active site of the class I and class II (aaRSs) shows that the nanodimensional tunnels are curved in opposite directions in the two classes. We investigated the origin of this difference using quantum mechanical computation of electrostatic potential (ESP) of substrates, surrounding residues and ions, using Atoms in Molecule (AIM) Theory and charge population analysis. We show that the difference is principally due to the variation in the spatial charge distribution of ATP in the two classes which correspond to extended and bent conformations of ATP. The present computation shows that the most feasible pathway for nucleophilic attack to alphaP is oppositely directed for class I and class II aaRSs. The available crystal structures show that the cognate AA is indeed located along the channel favorable for nucleophilic attack as predicted by the ESP analysis. It is also shown that the direction of the channel changes its orientation when the orientation of ATP is changed from extended to a bent like structure. We further used the AIM theory to confirm the direction of the approach of AA in each case and the results corroborate the results from the ESP analysis. The opposite curvatures of the active site nanospaces in class I and class II aaRSs are related with the influence of the charge distributions of the extended and bent conformations of ATP, respectively. The results of the computation of electrostatic potential by successive addition of active site residues show that their roles on the reaction are similar in both classes despite the difference in the organization of the

Changes in depression mediate the effects of AA attendance on alcohol use outcomes.

PubMed

Wilcox, Claire E; Tonigan, J Scott

2018-01-01

Depression may contribute to increased drinking in individuals with alcohol use disorder. Although Alcoholics Anonymous (AA) attendance predicts drinking reductions, there is conflicting information regarding the intermediary role played by reductions in depression. We explored whether AA attendance reduces depressive symptoms, the degree to which improvement in depression results in reductions in drinking, and in which subgroups these effects occur. 253 early AA affiliates (63% male) were recruited and assessed at baseline 3, 6, 9, 12, 18, and 24 months. Depression was measured using the Beck Depression Inventory (BDI) and was administered at baseline 3, 6, 12, 18, and 24 months. AA attendance and alcohol use outcomes were obtained with the Form 90. Mediation analyses were performed at early (3, 6, and 9 months) and late (12, 18, and 24 months) follow-up to investigate the degree to which reductions in depression mediated the effect of AA attendance on drinking, controlling for concurrent drinking. In addition, a series of moderated mediation analyses were performed using baseline depression severity as a moderator. At early follow-up, reductions in depression (6 months) mediated the effects of AA attendance (3 months) on later drinking (drinks per drinking day) (9 months) (b = -0.02, boot CI [-0.055, -0.0004]), controlling for drinking at 6 months. Baseline depression severity did not moderate the degree to which BDI mediated the effects of AA attendance on alcohol use (ps > .05). These findings provide further evidence that depression reduction is a mechanism by which AA attendance leads to reductions in alcohol use. Improving depression may help reduce alcohol use in individuals with AUD, and AA attendance may be an effective way to achieve that goal.

The NBS: Processing/Microstructure/Property Relationships in 2024 Aluminum Alloy Plates

NASA Technical Reports Server (NTRS)

Ives, L. K.; Swartzendruber, W. J.; Boettinger, W. J.; Rosen, M.; Ridder, S. D.

1983-01-01

As received plates of 2024 aluminum alloy were examined. Topics covered include: solidification segregation studies; microsegregation and macrosegregation in laboratory and commercially cast ingots; C-curves and nondestructive evaluation; time-temperature precipitation diagrams and the relationships between mechanical properties and NDE measurements; transmission electron microscopy studies; the relationship between microstructure and properties; ultrasonic characterization; eddy-current conductivity characterization; the study of aging process by means of dynamic eddy current measurements; and Heat flow-property predictions, property degradations due to improve quench from the solution heat treatment temperature.

Investigation of the intermolecular recognition mechanism between the E3 ubiquitin ligase Keap1 and substrate based on multiple substrates analysis.

PubMed

Jiang, Zheng-Yu; Xu, Li-Li; Lu, Meng-Chen; Pan, Yang; Huang, Hao-Ze; Zhang, Xiao-Jin; Sun, Hao-Peng; You, Qi-Dong

2014-12-01

E3 ubiquitin ligases are attractive drug targets due to their specificity to the ubiquitin machinery. However, the development of E3 ligase inhibitors has proven challenging for the fact that they must disrupt protein-protein interactions (PPIs). The E3 ligase involved in interactome provide new hope for the discovery of the E3 ligase inhibitors. These currently known natural binding partners of the E3 ligase can benefit the discovery of other unknown substrates and also the E3 ligase inhibitors. Herein, we present a novel strategy that using multiple substrates to elucidate the molecular recognition mechanism of E3 ubiquitin ligase. Molecular dynamics simulation, molecular mechanics-generalized born surface area (MM-GBSA) binding energy calculation and energy decomposition scheme were incorporated to evaluate the quantitative contributions of sub-pocket and per-residue to binding. In this case, Kelch-like ECH-associated protein-1 (Keap1), a substrate adaptor component of the Cullin-RING ubiquitin ligases complex, is applied for the investigation of how it recognize its substrates, especially Nrf2, a master regulator of the antioxidant response. By analyzing multiple substrates binding determinants, we found that both the polar sub-pockets (P1 and P2) and the nonpolar sub-pockets (P4 and P5) of Keap1 can make remarkable contributions to intermolecular interactions. This finding stresses the requirement for substrates to interact with the polar and nonpolar sub-pockets simultaneously. The results discussed in this paper not only show the binding determinants of the Keap1 substrates but also provide valuable implications for both Keap1 substrate discovery and PPI inhibitor design.

Investigation of the intermolecular recognition mechanism between the E3 ubiquitin ligase Keap1 and substrate based on multiple substrates analysis

NASA Astrophysics Data System (ADS)

Jiang, Zheng-Yu; Xu, Li-Li; Lu, Meng-Chen; Pan, Yang; Huang, Hao-Ze; Zhang, Xiao-Jin; Sun, Hao-Peng; You, Qi-Dong

2014-12-01

E3 ubiquitin ligases are attractive drug targets due to their specificity to the ubiquitin machinery. However, the development of E3 ligase inhibitors has proven challenging for the fact that they must disrupt protein-protein interactions (PPIs). The E3 ligase involved in interactome provide new hope for the discovery of the E3 ligase inhibitors. These currently known natural binding partners of the E3 ligase can benefit the discovery of other unknown substrates and also the E3 ligase inhibitors. Herein, we present a novel strategy that using multiple substrates to elucidate the molecular recognition mechanism of E3 ubiquitin ligase. Molecular dynamics simulation, molecular mechanics-generalized born surface area (MM-GBSA) binding energy calculation and energy decomposition scheme were incorporated to evaluate the quantitative contributions of sub-pocket and per-residue to binding. In this case, Kelch-like ECH-associated protein-1 (Keap1), a substrate adaptor component of the Cullin-RING ubiquitin ligases complex, is applied for the investigation of how it recognize its substrates, especially Nrf2, a master regulator of the antioxidant response. By analyzing multiple substrates binding determinants, we found that both the polar sub-pockets (P1 and P2) and the nonpolar sub-pockets (P4 and P5) of Keap1 can make remarkable contributions to intermolecular interactions. This finding stresses the requirement for substrates to interact with the polar and nonpolar sub-pockets simultaneously. The results discussed in this paper not only show the binding determinants of the Keap1 substrates but also provide valuable implications for both Keap1 substrate discovery and PPI inhibitor design.

Investigation of Hot Cracking Behavior in Transverse Mechanically Arc Oscillated Autogenous AA2014 T6 TIG Welds

NASA Astrophysics Data System (ADS)

Biradar, N. S.; Raman, R.