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Sample records for air brazing rab

  1. Silver-Copper Oxide Based Reactive Air Braze (RAB) for Joining Yttria-Stabilized Zirconia

    SciTech Connect

    Kim, Jin Yong Y.; Hardy, John S.; Weil, K. Scott

    2005-03-01

    We are investigating a new method of ceramic-to-metal joining, referred to as reactive air brazing (RAB), as a potential method of sealing ceramic components in high-temperature electrochemical devices. Sessile drop wetting experiments and joint strength testing were conducted using yttria stabilized zirconia (YSZ) substrates and CuO-Ag based air brazes. Results from our studies indicate that the wettability of the braze improves substantially with increasing CuO content, over a compositional range of 1 - 8 mol% CuO, which is accompanied by an increase in the bend strength of the corresponding brazed YSZ joint. The addition of a small amount of TiO2 (0.5 mol%) to the CuO-Ag braze further improves wettability due to the formation of a titanium zirconate reaction product along the braze/substrate interface. However, with one notable exception, the bend strength of these ternary braze joints remained nearly identical to those measured in comparable binary braze joints. SEM analysis conducted on the corresponding fracture surfaces indicated that in the binary braze joints the failure occurs primarily at the braze/YSZ interface. Similarly in the case of the the ternary, TiO2-doped brazes joint failure occurs predominantly along the interface between the braze filler metal and the underlying titanium zirconate reaction layer.

  2. Wetting and Mechanical Characteristics of the Reactive Air Braze for Yttria-Stabilized Zirconia (YSZ) Joining

    SciTech Connect

    Kim, Jin Yong Y.; Weil, K. Scott; Hardy, John S.

    2005-03-01

    Reactive air brazing (RAB) technique was developed as an effective alternative for the joining of complicated ceramic parts. The most important advantage of RAB over conventional active metal brazing is that joining operation of RAB technique can be conducted without using controlled atmosphere. It has been reported by us that the reactive component (copper) in the Ag-CuO braze is reactively to modify faying surfaces of alumina, improving the wettability with the oxide and potentially increasing bond strenth between braze and ceramics. In this work, the effects of CuO content on wetting behavior of Ag-CuO braze with yttria-stabilized zirconia (YSZ) substrates and the mechanical properties of brazed YSZ have been investigated. The results of this study to date will be discussed.

  3. Pd-modified Reactive Air Braze for Increased Melting Temperature

    SciTech Connect

    Hardy, John S.; Weil, K. Scott; Kim, Jin Yong Y.; Darsell, Jens T.

    2005-03-01

    Complex high temperature devices such as planar solid oxide fuel cell (pSOFC) stacks often require a two-step sealing process. For example, in pSOFC stacks the oxide ceramic fuel cell plates might be sealed into metallic support frames in one step. Then the frames with the fuel plates sealed to them would be joined together in a separate sealing step to form the fuel cell stack. In this case, the initial seal should have a sufficiently high solidus temperature that it will not begin to remelt at the sealing temperature of the material used for the subsequent sealing step. Previous experience has indicated that, when heated at a rate of 10°C/min, Ag-CuO reactive air braze (RAB) compositions have solidus and liquidus temperatures in the approximate range of 925 to 955°C. Therefore, compositionally modifying the original Ag-CuO braze with Pd-additions such that the solidus temperature of the new braze is between 1025 and 1050°C would provide two RAB compositions with a difference in melting points large enough to allow reactive air brazing of both sets of seals in the fuel cell stack. This study determines the appropriate ratio of Pd to Ag in RAB required to achieve a solidus in the desired range and discusses the wettability of the resulting Pd-Ag-CuO brazes on YSZ substrates. The interfacial microstructures and flexural strengths of Pd-Ag-CuO joints in YSZ will also be presented.

  4. Development of High-Temperature Air Braze Filler Metals for Use in Two-Step Joining and Sealing Processes

    SciTech Connect

    Hardy, John S.; Weil, K. Scott

    2006-11-02

    Reactive air brazing (RAB) creates metallic braze joints between oxide surfaces. RAB can be performed in air and can undergo subsequent operation at temperatures up to 900ºC in oxidizing environments. This makes RAB an attractive joining method for creating seals in solid oxide fuel cells (SOFCs) which operate at temperatures between 700 and 850ºC and comprise cathode materials which are intolerant of reducing environments. Many planar SOFC designs require a two-step sealing process. Thus the seal formed during the first step must not melt or soften at the firing temperature of the second step, otherwise its integrity could be compromised. The goal of this study is to investigate the effects of adding Pd to a Ag-CuO RAB to produce a braze composition with a melting temperature high enough that it might be used in the first step of a two-step sealing process with unmodified Ag-CuO RAB used in the second step, thereby making possible a two step reactive air brazing process. Yttria-stabilized zirconia (YSZ) is the substrate of choice in this study in order to simulate the initial sealing step in planar SOFCs which often involves sealing a yttrium-stabilized zirconia (YSZ) electrolyte to a metallic support frame. RAB compositions containing a 15 mol% Pd : 85 mol% Ag alloy with 8 mol% Cu added were found to provide the best combination of wettability, mechanical strength, and melting characteristics for brazing YSZ.

  5. Improved Wetting Characteristics in TiO2–Modified Ag-CuO Air Braze Filler Metals

    SciTech Connect

    Weil, K. Scott; Kim, Jin Yong; Hardy, John S.; Darsell, Jens T.

    2006-01-04

    A silver-based joining technique referred to as reactive air brazing (RAB) has been recently developed for joining high temperature structural ceramic components of the type used in high-temperature electrochemical devices. In prior work, it was found that additions of CuO to silver have a significant effect on the wettability and joint strength characteristics of the resulting braze on polycrystalline alumina substrates. More recently, it has been found that by adding as little as 0.5 mol % titania to these Ag-CuO brazes, the wettability of the RAB on alumina surfaces is further enhanced. The results of wettabilty measurements of Ag-CuO-TiO2 RAB compositions on alumina will be presented along with the microstructural characterization of Ag-CuO-TiO2 braze joints in alumina.

  6. Microstructure, Melting and Wetting Properties of Pd-Ag-CuO Air Braze on Alumina

    SciTech Connect

    Darsell, Jens T.; Hardy, John S.; Kim, Jin Yong Y.; Weil, K. Scott

    2004-05-01

    A new ceramic brazing technique, referred to as reactive air brazing (RAB), has recently been developed for potential applications in high temperature devices such as gas concentrators, solid oxide fuel cells, gas turbines, and combustion engines. At present, the technique utilizing a silver-copper oxide system is of great interest. The maximum operating temperature of this system, however, is limited by its eutectic temperature of ~935°C, although in practice the operating temperature should be limited to be even lower. An obvious strategy that can be employed to increase the maximum operating temperature of the braze material is to add a higher melting noble alloying element. In this paper, we report the effects of palladium addition on the thermodynamics of the Ag-CuO system and on the wetting properties of the resulting braze with respect to alumina

  7. Microstructure, Melting and Wetting Properties of Pd-Ag-CuO Air Braze on Alumina

    SciTech Connect

    Darsell, Jens T.; Hardy, John S.; Kim, Jin Yong Y.; Weil, K. Scott

    2005-03-19

    A reactive air brazing (RAB) technique utilizing silver-copper oxide (Ag-CuO) alloys has previously been developed for joining ceramics components used in high temperature devices ranging from oxygen separation membranes, gas turbines and combustion engines. The application of the Ag-CuO system as a brazing material is limited by its solidus and liquidus temperatures, which are known to be in the range of 935 C and 967 C. Some joined ceramic components may be used in devices, which require further processing steps, or may be used in applications, that exceed these temperatures. It has been found that the addition of palladium to the silver copper oxide system will increase solidus and liquidus temperatures of the resulting alloy. In our work, we are studying the effects of palladium addition on the wetting properties of Ag-CuO braze system on alumina. Quality of brazing is evaluated through microstructural analysis and bending strength of brazed joints created with alumina. The presentation will include processing, and characterization of Ag-CuO brazed system with and without palladium addition on alumina.

  8. Microstructural Development and Mechanical Properties for Reactive Air Brazing of ZTA to Ni Alloys using Ag-CuO Braze Alloys

    SciTech Connect

    Prevost, Erica; DeMarco, A.Joseph; MacMichael, Beth; Joshi, Vineet V.; Meier, Alan; Hoffman, John W.; Walker, William J.

    2014-12-01

    Reactive air brazing (RAB) is a potential joining technique to join metal alloys to ceramics for a variety of applications. In the current study, nickel (Ni) alloys were heat treated to form an oxide layer prior to RAB joining to zirconia toughened alumina (ZTA). The Ni alloys evaluated were Nicrofer 6025 HT, Inconel 600, Inconel 693, Haynes 214 and Inconel 601. The ZTA studied had compositions of 0 to 15 wt% zirconia and 0 to 14 wt% glass. Four point-bend tests were performed to evaluate the joint strength of ZTA/ZTA and ZTA/nickel alloys brazed with Ag-2wt% CuO braze alloys. It was determined that the joint strength is not a function of the ZTA composition, but that the strength is a strong function of the chemistry and microstructure of the oxide layer formed on the nickel alloy. It was determined that an increase in the aluminum content of the Ni alloy resulted in an increase of the thickness of alumina in the oxide layer and was directly proportional to the bond strength with the exception of Inconel 601 which exhibited relatively high joint strengths even though it had a relatively low aluminum content.

  9. Diffusion barriers in modified air brazes

    DOEpatents

    Weil, Kenneth Scott; Hardy, John S.; Kim, Jin Yong; Choi, Jung-Pyung

    2010-04-06

    A method for joining two ceramic parts, or a ceramic part and a metal part, and the joint formed thereby. The method provides two or more parts, a braze consisting of a mixture of copper oxide and silver, a diffusion barrier, and then heats the braze for a time and at a temperature sufficient to form the braze into a bond holding the two or more parts together. The diffusion barrier is an oxidizable metal that forms either a homogeneous component of the braze, a heterogeneous component of the braze, a separate layer bordering the braze, or combinations thereof. The oxidizable metal is selected from the group Al, Mg, Cr, Si, Ni, Co, Mn, Ti, Zr, Hf, Pt, Pd, Au, lanthanides, and combinations thereof.

  10. Diffusion barriers in modified air brazes

    DOEpatents

    Weil, Kenneth Scott; Hardy, John S; Kim, Jin Yong; Choi, Jung-Pyung

    2013-04-23

    A method for joining two ceramic parts, or a ceramic part and a metal part, and the joint formed thereby. The method provides two or more parts, a braze consisting of a mixture of copper oxide and silver, a diffusion barrier, and then heats the braze for a time and at a temperature sufficient to form the braze into a bond holding the two or more parts together. The diffusion barrier is an oxidizable metal that forms either a homogeneous component of the braze, a heterogeneous component of the braze, a separate layer bordering the braze, or combinations thereof. The oxidizable metal is selected from the group Al, Mg, Cr, Si, Ni, Co, Mn, Ti, Zr, Hf, Pt, Pd, Au, lanthanides, and combinations thereof.

  11. Protecting brazing furnaces from air leaks

    NASA Technical Reports Server (NTRS)

    Armenoff, C. T.; Mckown, R. D.

    1980-01-01

    Inexpensive inert-atmosphere shielding protects vacuum brazing-furnace components that are likely to spring leak. Pipefittings, gages, and valves are encased in transparent plastic shroud inflated with argon. If leak develops, harmless argon will enter vacuum chamber, making it possible to finish ongoing brazing or heat treatment before shutting down for repair.

  12. The Effect of Braze Interlayer Thickness on the Mechanical Strength of Alumina Brazed with Ag-CuO Braze Alloys

    SciTech Connect

    Erskine, Kevin M.; Meier, Alan; Joshi, Vineet V.; Pilgrim, Steven M.

    2014-12-01

    The effect of braze interlayer thickness on the strength of alumina brazed with silver-copper oxide reactive air braze (RAB) alloys was evaluated using a four point bend test configuration. The brazed samples had an average fracture strength of 180 MPa or approximately 60 percent of the average monolithic alumina strength. The joint strength values obtained exceeded the yield strength and ultimate tensile strength of the silver interlayer indicating strong ceramic to metal adhesion and the development of a triaxial stress state in the braze interlayer. The average fracture strength was relatively constant (190 ± 60 MPa) in the thickness range of 0.030 mm to 0.230 mm for all test conditions. The braze fracture strength then decreased down to 100 ± 30 MPa as the braze thickness increased from 0.230 mm to 0.430 mm indicating a loss of triaxial constraint with increasing interlayer thickness. In addition, four different fracture modes were observed.

  13. Brazing.

    SciTech Connect

    Cadden, Charles H.

    2005-04-01

    The use of a lower-melting-point molten metal to join metallic components is perhaps the earliest example of processing which employs metallurgical bonding principles, having roots as far back as 4200 BC (Peaslee 2003). More than 6000 years later, brazing occupies a prominent position in our suite of joining processes where it offers cost and/or performance advantages in the fabrication of many structures. More precisely, brazing can be described as the use of a molten filler metal to wet the closely fitting faying surfaces of a joint, leading to formation of metallurgical bonds between the filler metal and substrates. Historically, brazing processes employ filler metals whose solidus temperature exceeds 723 K, as opposed to soldering processes which use lower-melting-point temperature filler materials. In the past several decades, technological advances have facilitated a broadening of applications for brazing while simultaneously contradicting some of the traditional perceptions of the process. However, many of those tenets remain appropriate for the majority of brazing processes and products. Accordingly, this article provides a brief description of traditional brazing and some important factors to be considered when designing and producing brazed structures. An additional section describes the technical advances in the field.

  14. Joining Mixed Conducting Oxides Using an Air-Fired Electrically Conductive Braze

    SciTech Connect

    Hardy, John S.; Kim, Jin Yong Y.; Weil, K. Scott

    2004-10-01

    Due to their mixed oxygen ion and electron conducting properties, ceramics such as lanthanum strontium cobalt ferrites (LSCF) are attractive materials for use in active electrochemical devices such as solid oxide fuel cells (SOFC) and oxygen separation membranes. However, to take full advantage of the unique properties of these materials, reliable joining techniques need to be developed. If such a joining technique yields a ceramic-to-metal junction that is also electrically conductive, the hermetic seals in the device could provide the added function of either drawing current from the mixed conducting oxide, in the case of SOFC applications, or carrying it to the oxide to initate ionic conduction, in the case of oxygen separation and electrocatalysis applications. This would greatly reduce the need for complex interconnect design, thereby simplifying one of the major challenges faced in SOFC development. A process referred to as reactive air brazing (RAB) has been developed in which firing a Ag-CuO filler material in air creates a functional ceramic-to-metal junction, in which the silver-based matrix of the braze affords both metallic ductility and conductivity in the joint. Investigating a range of Ag-CuO alloy combinations determined that compositions containing between 1.4 and 16 mol% CuO appear to offer the best combination of wettability, joint strength, and electrical conductivity.

  15. The effect of palladium additions on the solidus/liquidus temperatures and wetting properties of Ag-CuO based air brazes

    SciTech Connect

    Darsell, Jens T.; Weil, K. Scott

    2007-01-01

    A new ceramic brazing technique referred to as reactive air brazing (RAB) has recently been developed for potential applications in high temperature devices such as gas concentrators, solid oxide fuel cells, gas turbines, and combustion engines. At present, the technique utilizing a silver-copper oxide system is of great interest. The maximum operating temperature of this system is limited by its eutectic temperature of ~945°C, although in practice the operating temperature will need to be lower. An obvious strategy that can be employed to increase the maximum operating temperature of the braze material is to add a higher melting noble alloying element. In this paper, we report the effects of palladium addition on the melting characteristics of the Ag-CuO system and on the wetting properties of the resulting braze with respect to alumina. It was found that the addition of Pd will cause an increase in the melting temperature of the Ag-CuO braze but possibly at a sacrifice of wetting properties depending on composition.

  16. INVESTIGATION OF A NOVEL AIR BRAZING COMPOSITION FOR HIGH-TEMPERATURE, OXIDATION-RESISTANT CERAMIC JOINING

    SciTech Connect

    Weil, K. Scott; Hardy, John S.; Darsell, Jens T.

    2004-01-30

    One of the challenges in developing a useful ceramic joining technique is in producing a joint that offers good strength under high temperature and highly oxidizing operating conditions. Unfortunately many of the commercially available active metal ceramic brazing alloys exhibit oxidation behaviors which are unacceptable for use in a high temperature application. We have developed a new approach to ceramic brazing, referred to as air brazing, that employs an oxide wetting agent dissolved in a molten noble metal solvent, in this case CuO in Ag, such that acceptable wetting behavior occurs on a number of ceramic substrates. In an effort to explore how to increase the operating temperature of this type of braze, we have investigated the effect of ternary palladium additions on the wetting characteristics of our standard Ag-CuO air braze composition

  17. Ag-Al based air braze for high temperature electrochemical devices

    SciTech Connect

    Kim, Jin Yong Y.; Hardy, John S.; Weil, K. Scott

    2007-11-01

    Silver-aluminum based air brazing was attempted using an in-situ alloying and brazing process. In this process, layers of foils of aluminum and silver were laid up between alumina plates in alternating fashion to achieve three target compositions representing Ag, Ag3Al, and Ag2Al phases. Each alloy composition revealed different microstructure, mechanical properties and fracture mechanisms. Joints brazed with foils containing 9.8 at% Al formed a long continuous layer parallel to the direction of the original aluminum foil. The fracture occurred through the interface between this long alumina layer and the braze filler, resulting in low bend strength (6 ~ 12 MPa). Joints containing 26.5 at% Al in the braze filler metal experienced the series of phase transformations, leading to cracks in as-brazed specimens. The fracture initiated through these pre-existing cracks, thus the joint strength observed in these specimens was extremely low. The joints prepared using foils with 35.1 at% Al exhibited a good interface even though interfacial alumina particles formed during air brazing. Crack propagation occurred through the interface between the alumina substrate and in-situ formed interfacial alumina particles or directly through these particles and the best bend strength (46 ~ 52 MPa) among Al-added braze compositions was achieved.

  18. High temperature tolerance of the silver-copper oxide braze in reducing and oxidizing atmospheres

    SciTech Connect

    Kim, Jin Yong Y.; Hardy, John S.; Weil, K. Scott

    2006-06-01

    Silver-copper oxide based reactive air brazing (RAB) technique was developed as an alternative technique for joining complex-shaped ceramic parts. To examine the feasibility of this braze for various high temperature applications, brazed alumina joints were exposed to oxidizing and reducing atmospheres at high temperature. Brazed joints, which were exposed to 800ºC in air for 100 h, maintained good bend strength similar to the as-brazed samples. Microstructural analysis also revealed no significant change after exposing the joints to the oxidizing atmosphere at high temperature. This result indicate the excellent high-temperature tolerance of the Ag-CuO based braze in oxidizing atmosphere. On the other hand, heat treatment of the brazed alumina joints in hydrogen at 800°C for 100 h resulted in significant decrease in bend strength. SEM analysis on fracture surfaces showed that the main fracture mechanism of the samples exposed to the reducing atmosphere was the debonding between the braze and the alumina substrate. This result indicates that the bond strength of the braze/alumina interface is sensitive to the atmosphere where the brazed joint is exposed. CuO in the braze was also reduced to Cu and diffused into the Ag matrix. This reduction of CuO created the pores at the interface as well as in the braze matrix where CuO was originally located, especially at a high CuO content.

  19. AN INVESTIGATION OF WETTABILITY, AND MICROSTRUCTURE IN ALUMINA JOINTS BRAZED WITH Ag-CuO-TiO2

    SciTech Connect

    Darsell, Jens T.; Hardy, John S.; Kim, Jin Yong Y.; Weil, K. Scott

    2004-04-06

    A silver-based joining technique referred to as reactive air brazing (RAB) has been recently developed for joining high temperature structural ceramic components of the type used in gas turbines, combustion engines, heat exchangers, and burners. It was found that additions of CuO to silver have a significant effect on the wettability and joint strength characteristics of the resulting braze on polycrystalline alumina substrates. More recently, it has been found that by adding as little as 0.5 mol% titania to these Ag-CuO brazes, the wettability of the RAB on alumina surfaces is further enhanced. The results of wettabilty measurements of Ag-CuO-TiO2 RAB compositions on alumina will be presented along with the microstructural characterization of Ag-CuO-TiO2 braze joints in alumina.

  20. Brazing retort manifold design concept may minimize air contamination and enhance uniform gas flow

    NASA Technical Reports Server (NTRS)

    Ruppe, E. P.

    1966-01-01

    Brazing retort manifold minimizes air contamination, prevents gas entrapment during purging, and provides uniform gas flow into the retort bell. The manifold is easily cleaned and turbulence within the bell is minimized because all manifold construction lies outside the main enclosure.

  1. Dual atmosphere tolerance of Ag-CuO based air braze

    SciTech Connect

    Kim, Jin Yong Y.; Hardy, John S.; Weil, K. Scott

    2007-11-01

    Recently, a new braze filler metal based on the silver-copper oxide system was developed for use in sealing high-temperature, solid-state electrochemical devices such as solid oxide fuel cells. One of the concerns regarding the viability of this joining technique is the long-term stability of silver-based alloys under a high-temperature, dual oxidizing/reducing gas environment. This paper reports on an initial series of exposure experiments that were conducted to characterize the effects of (1) filler metal composition, (2) brazing temperature, and (3) exposure time on the microstructural stability of Ag-CuO brazed Al2O3/Al2O3 joints under a prototypic operating environment for an intermediate temperature solid oxide fuel cell stack. In general joints exposed simultaneously to air on one side and hydrogen on the other for short periods of time at 800°C (100 hrs) showed no signs of degradation with respect to hermeticity or joint microstructure. Samples exposed for longer periods of time (1000 hrs) displayed some internal porosity, which extends approximately halfway across the joint and is not interconnected. Little effect of the filler metal’s composition on its tolerance to dual atmosphere exposure was observed. However brazing temperature was found to have a measurable effect. Higher brazing temperature leads to a more extensive formation of an interfacial reaction phase, copper aluminate, which tends to tie up some of the free CuO in the filler metal and minimize the formation of porosity in the air brazed joints during long-term, dual-atmosphere exposure. The effect is due to the greater chemical stability of the copper aluminate relative to copper oxide.

  2. Mechanical properties and dual atmosphere tolerance of Ag-Al based braze

    SciTech Connect

    Kim, Jin Yong; Choi, Jung-Pyung; Weil, K. Scott

    2008-03-14

    Reactive air brazing (RAB) based on the silver-copper oxide system was recently developed for use in sealing high-temperature electrochemical devices such as solid oxide fuel cells. One of the concerns regarding the viability of this joining technique is the long-term stability of silver-based alloys under a high-temperature, dual oxidizing/reducing gas environment. One possible solution to improve the dual atmosphere tolerance of the silver-based system is the addition of elements which can preferentially react with oxygen over hydrogen and minimize the pore formation caused by the reaction of oxygen with hydrogen in the silver matrix. In this paper, the effects of aluminum addition into silver-based air braze filler materials on microstructure, mechanical properties, and high temperature dual atmosphere tolerance were investigated using foils and pastes of aluminum-added braze filler materials. Joints brazed with binary Ag-Al braze foils containing more than 2 at% of Al retained a metallic form of aluminum in the metallic braze filler matrix after brazing at 1000°C in air. The flexural strength of joints prepared with binary Ag-Al braze foils decreased with increase in Al content due to the formation of interfacial aluminum oxide. The existence of metallic aluminum in the braze filler matrix, however, enhanced the high temperature dual atmosphere tolerance of the silver-based braze filler, showing smaller size of porosity after dual reducing/oxidizing atmosphere tests at 800°C for 1000 hrs. The Binary and ternary braze pastes based on the Ag-Al(-Cu) system were also tried as a sealant. Alumina joints brazed with these pastes showed increase in flexural strength with Cu content. However, a braze filler containing 5 at% Al and 8 at% Cu possessed nearly no metallic aluminum in the braze filler matrix after brazing, while the as-brazed sample prepared using a binary braze filler with 5 at% Al kept some metallic Al in the braze matrix. Thus, the addition of copper

  3. An Interfacial Characterization of Pd-Ag-CuO Reactive Air Joints with Alumina

    SciTech Connect

    Weil, K. Scott; Darsell, Jens T.

    2005-05-26

    A new ceramic brazing technique referred to as reactive air brazing (RAB) is currently under development that has potential applications in high temperature devices such as gas concentrators, solid oxide fuel cells, gas turbines, and combustion engines. Currently, the technique utilizes a silver-copper oxide system. We have previously increased the operating temperature of this system by adding palladium. In this paper, we report the effects of palladium addition on the wetting properties of the resulting braze relative to alumina. Specifically, we will discuss the apparent correlation between braze wettability and interfacial microstructure between the braze and alumina, both with and without palladium additions.

  4. Investigation of Pd-Modified Ag-CuO Air Braze Filler Metals

    SciTech Connect

    Weil, K. Scott; Kim, Jin Yong; Hardy, John S.; Darsell, Jens T.

    2006-01-01

    This paper reports on the effects of palladium on the liquidus/solidus temperatures and wetting behavior of a series of Ag-CuOx air braze filler metals. Currently, the maximum operating temperature of the Ag-CuOx system is limited by its eutectic temperature of ~935°C. One strategy to increase the maximum operational temperature of this family of filler metals is to add a higher melting noble alloying element. In the current study, we examined the effects of palladium additions on the melting characteristics of the Ag-CuO materials and the wetting properties of the resulting air braze filler metals with respect to alumina. It was found that while the addition of Pd causes the anticipated increase in the melting temperature, it does so at a sacrifice in wetting properties. The extent of both effects and therefore the opportunity to trade-off the two properties in order to develop an optimized higher temperature air braze depends on concentrations of both the palladium and copper oxide.

  5. WETTING AND REACTIVE AIR BRAZING OF BSCF FOR OXYGEN SEPARATION DEVICES

    SciTech Connect

    LaDouceur, Richard M.; Meier, Alan; Joshi, Vineet V.

    2014-10-13

    Reactive air brazes Ag-CuO and Ag-V2O5 were evaluated for brazing Ba0.5Sr0.5Co0.8Fe0.2O(3-δ) (BSCF). BSCF has been determined in previous work to have the highest potential mixed ionic/electronic conducting (MIEC) ceramic material based on the design and oxygen flux requirements of an oxy-fuel plant such as an integrated gasification combined cycle (IGCC) used to facilitate high-efficiency carbon capture. Apparent contact angles were observed for Ag-CuO and Ag-V2O5 mixtures at 1000 °C for isothermal hold times of 0, 10, 30, and 60 minutes. Wetting apparent contact angles (θ<90°) were obtained for 1%, 2%, and 5% Ag-CuO and Ag-V2O5 mixtures, with the apparent contact angles between 74° and 78° for all compositions and furnace dwell times. Preliminary microstructural analysis indicates that two different interfacial reactions are occurring: Ag-CuO interfacial microstructures revealed the same dissolution of copper oxide into the BSCF matrix to form copper-cobalt-oxygen rich dissolution products along the BSCF grain boundaries and Ag-V2O5 interfacial microstructures revealed the infiltration and replacement of cobalt and iron with vanadium and silver filling pores in the BSCF microstructure. The Ag-V2O5 interfacial reaction product layer was measured to be significantly thinner than the Ag-CuO reaction product layer. Using a fully articulated four point flexural bend test fixture, the flexural fracture strength for BSCF was determined to be 95 ± 33 MPa. The fracture strength will be used to ascertain the success of the reactive air braze alloys. Based on these results, brazes were fabricated and mechanically tested to begin to optimize the brazing parameters for this system. Ag-2.5% CuO braze alloy with a 2.5 minute thermal cycle achieved a hermetic seal with a joint flexural strength of 34 ± 15 MPa and Ag-1% V2O5 with a 30 minute thermal cycle had a joint flexural strength of 20 ± 15 MPa.

  6. Air Brazing: A New Method of Ceramic-Ceramic and Ceramic-Metal Joining

    SciTech Connect

    Weil, K. Scott; Darsell, Jens T.; Kim, Jin Yong

    2011-10-01

    A new method of ceramic-ceramic and ceramic-metal joining has emerged over the past several years. Referred to as air brazing, the technique was originally designed and developed for use in fabricating high-temperature solid-state electrochemical devices such as planar SOFCs and oxygen and hydrogen concentrators. The primary advantage of air brazing is that a predominantly metallic joint can be formed directly in air without need of an inert cover gas or the use of surface reactive fluxes. The resulting bond is hermetic, offers excellent room temperature strength, and is inherently resistant to oxidation at high temperature. The key to developing a successful filler metal composition for air brazing is to identify a metal oxide wetting agent that is mutually soluble in a molten noble metal solvent. One particular oxide-metal combination that appears readily suited for this purpose is CuOx-Ag, a system originally of interest in the development of silver clad cuprate-based superconductors. Studies of the equilibrium phases studies in this system indicate that there are two invariant points in the pseudobinary CuOx-Ag phase diagram around which new braze compositions can be developed: 1) a monotectic reaction at 969±1°C, where CuO and a Ag-rich liquid L1 coexist with a second CuOx-rich liquid phase L2 at a composition of xAg/(xAg + xCu) = 0.10±0.03 Ag and 2) a eutectic reaction at 942±1°C, where CuO and Ag coexist with L1 at a composition of xAg/(xAg + xCu) = 0.99±0.005. Specifically, near-eutectic Ag-CuO filler metal compositions have shown good promise in joining electrochemically active ceramics such as yttria-stabilized zirconia, lanthanum strontium manganite, and barium strontium cobalt ferrite, as well as alumina and magnesia. More recently it has been found that various ternary additions can further improve the wetting characteristics of these filler metals, increase their potential operating temperatures, and/or increase the resulting strength of the

  7. Brazing technique

    DOEpatents

    Harast, Donald G.

    1986-01-01

    A method of brazing comprises sand blasting the surfaces of the components to be brazed with particles of a brazing material to clean the surfaces and to embed brazing material in the surfaces, applying the brazing material to the surfaces, and heating the brazing material to form a brazement between the components.

  8. Investigation of Pd-Modified Ag-CuO Air Braze Filler Metals

    SciTech Connect

    Darsell, Jens T.; Hardy, John S.; Kim, Jin Yong; Weil, K. Scott

    2006-01-10

    Palladium was added as a ternary component to a series of silver - copper oxide alloys in an effort to increase the use temperature of these materials for potential ceramic air brazing applications. Large portions of the silver component of the Ag-CuO system were substituted by palladium forming the following alloys: (100-y)[(100-z)Pd - (z)Ag] - (y)CuOx where y = 0 - 34 mol% CuOx, z = 50 - 100 mol% silver, and x = 0, 0.5, and 1, denoting copper metal, Cu2O, or CuO. From differential scanning calorimetry, it was determined that the addition of palladium causes an increase in the solidus and liquidus temperatures of the resulting Pd-Ag-CuO brazes. In general, the liquidus was found to increase by approximately 220°C for the (100-y)(25Pd - 75Ag) - (z)CuOx filler metal compositions relative to comparable Ag-CuOx alloys. Likewise, the solidus was found to increase for these alloys, respectively by 185°C and 60°C, respectively for CuOx contents of y = 0 - 1mol% and 4 - 10 mol%. For the (100-y)(50Pd - 50Ag) - (y)CuOx alloys, the solidus increased between 280 - 390°C over a copper oxide compositional range of x = 0 to 8 mol%. It was determined from sessile drop experiments that palladium causes an increase in the wetting angle for all of the samples tested. Alloy compositions of (100-y)(25Pd - 75Ag) - (y)CuOx displayed increased wetting angles of 5-20° relative to comparable binary compositions. (100-y)(50Pd - 50Ag) - (y)CuOx alloys exhibited an increase in contact angle of 10-60° and compositions containing less than 10 mol% CuOx were not able to wet the substrate. Scanning electron microscopy indicates that the microstructure of the braze consists of Ag-Pd solid solution with CuOx precipitates. In general, a reaction layer consisting of CuAlO2 forms adjacent to the alumina substrate. However, the formation of this layer is apparently hindered by the addition of large amounts of palladium, causing poor wetting behavior, as denoted by substantial porosity found along

  9. Atomic structure and thermophysical properties of molten silver-copper oxide air braze alloys

    NASA Astrophysics Data System (ADS)

    Hardy, John Steven

    The Ag-CuOx materials system is the basis for a family of filler alloys used in a recently developed ceramic-metal joining technique referred to as air brazing, which is a brazing process that can be carried out in ambient air rather than under the vacuum or inert to reducing gas conditions required for conventional brazing methods. This research was conducted to elucidate the atomic coordination and selected thermophysical properties of these materials as a function of temperature when they are in the salient liquid state in air, since this is when the critical steps of wetting and spreading occur in the joining process. A series of alloys was selected spanning the entire length of the phase diagram including the pure end members, Ag and CuOx; alloys that form the two constituent single phase liquids; and alloys for which the two liquid phases coexist in the miscibility gap of the phase diagram. The oxygen content of the liquid alloys in air was measured using thermogravimetry. The oxidative weight gain of 99.999% pure metallic precursors was measured while simultaneously accounting for the concurrent silver volatility using a method that was developed in the course of the study. The surface tension and mass density were measured using the maximum bubble pressure method. The number density was calculated based on the information gained from the oxygen content and mass density measurements. For compositions that were amenable to laser heating, containerless high energy x-ray scattering measurements of the liquid atomic coordination were performed using a synchrotron beamline, an aerodynamic levitator, and laser heating. For the remaining compositions x-ray scattering measurements were performed in a beamline-compatible furnace. The two liquid phases that form in this materials system have distinct atomic coordinations characterized by an average of nearly two-fold coordinated ionic metal-oxygen pairs in the CuOx-rich liquid and nearly eight-fold coordinated atomic

  10. Improved Wetting of Mixed Ionic/Electronic Conductors Used in Electrochemical Devices with Ternary Reactive Air Braze Filler Metals

    SciTech Connect

    Hardy, John S; Kim, Jin Yong Y; Thomsen, Ed C; Weil, K Scott

    2007-01-19

    This paper reports on the wetting behavior, reactivity, and long-term electrical conductance of a series of ternary filler metals being considered for brazing lanthanum strontium cobalt ferrite (LSCF) based oxygen separation membranes. Mixed ionic/electronic conducting perovskite oxides such as LSCF and various doped barium cerates are currently being considered for use in high-temperature electrochemical devices such as oxygen and hydrogen concentrators and solid oxide fuel cells. However to take full advantage of the unique properties of these materials, reliable joining techniques need to be developed. Furthermore, if the proposed joining technique were to yield a hermetic ceramic-to-metal junction that was also electrically conductive, it would additionally benefit the device by allowing current to be drawn from or carried to the electrochemically active mixed conducting oxide component without requiring an separate current collector. A newly developed brazing technique known as air brazing is one such method of joining. In its present form, air brazing uses a silver-copper oxide based filler metal that can be melted directly in air to form a compliant joint that is electrically conductive. Recently, it has been shown that the addition of titania can enhance the wetting behavior of Ag-CuO filler metals on alumina. Here the effect of this wetting agent on the surface wettability, long-term electrical resistance at 750°C, and reactivity with La0.6Sr0.4Co0.2Fe0.8O3- (LSCF-6428 or LSCF) substrates is discussed.

  11. Phase Evolution in the Pd-Ag-CuO Air Braze Filler Metal Alloy System

    SciTech Connect

    Darsell, Jens T.; Weil, K. Scott

    2006-08-01

    Palladium was added as a ternary component to a series of copper oxide-silver alloys in an effort to increase the use temperature of these materials for potential ceramic air brazing applications. Phase equilibria in the ternary Pd-Ag-CuO system were investigated via differential scanning calorimetry (DSC) and a series of quenching experiments. Presented here are the latest findings on this system and a construction of the corresponding ternary phase diagram for low-to-moderate additions of palladium. The analysis included samples with higher palladium additions than were studied in the past, as well as an analysis of the composition-temperature trends in the Ag-CuO miscibility gap with palladium addition. It was found that the addition of palladium increases the solidus and liquidus and caused three phase zones to appear as expected by the phase rule. Furthermore, the palladium additions cause the miscibility gap boundary extending from the former binary eutectic to shift to lower silver-to-copper ratios.

  12. BRAZING OF POROUS ALUMINA TO MONOLITHIC ALUMINA WITH Ag-CuO and Ag-V2O5 ALLOYS

    SciTech Connect

    Lamb, M. C.; Camardello, Sam J.; Meier, Alan; Weil, K. Scott; Hardy, John S.

    2005-01-31

    The feasibility of joining porous alumina (Al{sub 2}O{sub 3}) bodies to monolithic Al{sub 2}O{sub 3} using Ag-CuO and Ag-V{sub 2}O{sub 5} alloys via reactive air brazing (RAB) was examined for a nanoporous filter application. Brazing for these systems is complicated by the conflicting requirements of satisfactory wetting to fill the braze gap, while minimizing the infiltration of the porous body. By varying the firing time, temperature, and initial powder size, porous bodies with a range of pore microstructures were fabricated. The wettability was evaluated via sessile drop testing on monolithic substrates and porous body infiltration. Porous Al{sub 2}O{sub 3}/monolithic Al{sub 2}O{sub 3} brazed samples were fabricated, and the microstructures were evaluated. Both systems exhibited satisfactory wetting for brazing, but two unique types of brazing behavior were observed. In the Ag-CuO system, the braze alloy infiltrated a short distance into the porous body. For these systems, the microstructures indicated satisfactory filling of the brazed gap and a sound joint regardless of the processing conditions. The Ag-V{sub 2}O{sub 5} alloys brazed joints exhibited a strong dependence on the amount of V{sub 2}O{sub 5} available. For Ag-V{sub 2}O{sub 5} alloys with large V{sub 2}O{sub 5} additions, the braze alloy aggressively infiltrated the porous body and significantly depleted the Ag from the braze region resulting in poor bonding and large gaps within the joint. With small additions of V{sub 2}O{sub 5}, the Ag infiltrated the porous body until the V{sub 2}O{sub 5} was exhausted and the Ag remaining at the braze interlayer bonded with the Al{sub 2}O{sub 3}. Based on these results, the Ag-CuO alloys have the best potential for brazing porous Al{sub 2}O{sub 3} to monolithic Al{sub 2}O{sub 3}.

  13. Improved Wetting Characteristics in TiO2–Modified Ag-CuO Air Braze Filler Metals

    SciTech Connect

    Weil, K. Scott; Kim, Jin Yong Y.; Hardy, John S.; Darsell, Jens T.

    2006-01-01

    In this paper we report on the results of a series of sessile drop experiments designed to examine the effect of TiO2 on the wetting behavior of Ag-CuO air braze filler metals. It was found that TiO2 concentrations as small as 0.5 mol% can significantly decrease the contact angle of Ag-CuO on alumina over a compositional range of 1 – 34mol% CuO. The effect appears to maximize at a copper oxide concentration of ~4 mol% CuO regardless of the titania content.

  14. BRAZING ALLOYS

    DOEpatents

    Donnelly, R.G.; Gilliland, R.G.; Slaughter, G.M.

    1962-02-20

    A brazing alloy is described which, in the molten state, is characterized by excellent wettability and flowability and is capable of forming a corrosion-resistant brazed joint. At least one component of said joint is graphite and the other component is a corrosion-resistant refractory metal. The brazing alloy consists essentially of 40 to 90 wt % of gold, 5 to 35 wt% of nickel, and 1 to 45 wt% of tantalum. (AEC)

  15. Braze joints of dispersion strengthened copper

    NASA Astrophysics Data System (ADS)

    Chen, Shaofeng; Bao, Tingshan; Chin, Bryan A.

    1996-10-01

    Prior attempts to join alumina dispersion strengthened alloys (GlidCop) to itself, stainless steel (SS) and other materials have been less than successful. Typically these joints show low strength and poor ductility due to the segregation of alumina particles at the braze interface. This paper presents the results of a study undertaken to develop low activation brazing alloys, fluxes and processes for vacuum, argon and air environments to join alumina dispersion strengthened copper alloys to 304 SS. Vacuum and argon experiments were conducted using a Gleeble resistance heated system. Air brazing experiments were conducted in a resistance heated furnace with active fluxes. A reactive braze alloy composition has been developed which eliminates the segregation of Al 2O 3 particles at the braze interface and produces joints with excellent strength and ductility. Wetting and mechanical properties of the braze alloys were measured. Tensile properties, microstructure, microhardness and measurements of the compositional segregation across the braze joint are discussed.

  16. Development of Brazing Technology for Use in High- Temperature Gas Separation Equipment

    SciTech Connect

    Weil, K.S.; Hardy, J.S.; Kim, J.Y.

    2003-04-23

    -to-metal braze and consequently lead to the development of the novel reactive air brazing (RAB) concept. The goal in RAB is to reactively modify one or both oxide faying surfaces with an oxide compound dissolved in a molten noble metal alloy such that the newly formed surface is readily wetted by the remaining liquid filler material. In many respects, this concept is similar to active metal brazing, except that joining can be conducted in air and the final joint will be resistant to oxidation at high temperature. Potentially, there are a number of metal oxide-noble metal systems that can be considered for RAB, including Ag-CuO, Ag-V2O5, and Pt-Nb2O5. Our current interest is in determining whether the Ag-CuO system is suitable for air brazing functional ceramic-to-metal joints such as those needed in practical electrochemical devices. In a series of studies, the wetting behavior of the Ag-CuO braze was investigated with respect to a number of potential hydrogen separation, oxygen separation, and fuel cell electrolyte membrane materials and heat resistant metal systems, including: alumina, (La0.6Sr0.4)(Co0.2Fe0.8)O3, (La0.8Sr0.2)FeO3, YSZ, fecralloy, and Crofer-22APU. Selected findings from these studies as well as from our work on joint strength and durability during high-temperature exposure testing will be discussed.

  17. Overwhelming reaction enhanced by ultrasonics during brazing of alumina to copper in air by Zn-14Al hypereutectic filler.

    PubMed

    Ji, Hongjun; Chen, Hao; Li, Mingyu

    2017-03-01

    The ultrasonic-assisted brazing of α-alumina to copper was achieved in air without flux using Zn-14wt%Al hypereutectic filler at 753K within tens of seconds. The effects of ultrasonic time on the microstructures and mechanical properties of joints were investigated. In the joint interlayer, large amounts of intermetallic phases consisted of binary CuZn5 embedded by many ternary Al4.2Cu3.2Zn0.7 particles were formed. At the ceramic interface, newly formed crystalline Al2O3 aggregated. At the Cu interface, acoustic corrosion on the copper resulted in depriving the surface oxides and forming many pits on its surface, which provided saturated Cu in the melted filler alloys during the brazing. The ultrasonic vibrations had distinct effects on the metallurgical reactions of the joints, resulting in intermetallic-phase-filled composite joints with shear strength of 66MPa. The overgrowth of intermetallic compounds, the newly formed crystalline alumina, and the acoustic pits was probably ascribed to the ultrasonic effects.

  18. Laser Brazing of High Temperature Braze Alloy

    NASA Technical Reports Server (NTRS)

    Gao, Y. P.; Seaman, R. F.; McQuillan, T. J.; Martiens, R. F.

    2000-01-01

    The Space Shuttle Main Engine (SSME) consists of 1080 conical tubes, which are furnace brazed themselves, manifolds, and surrounding structural jacket making almost four miles of braze joints. Subsequent furnace braze cycles are performed due to localized braze voids between the coolant tubes. SSME nozzle experiences extremely high heat flux (180 mW/sq m) during hot fire. Braze voids between coolant tubes may result in hot combustion gas escape causing jacket bulges. The nozzle can be disqualified for flight or result in mission failure if the braze voids exceed the limits. Localized braze processes were considered to eliminate braze voids, however, damage to the parent materials often prohibited use of such process. Being the only manned flight reusable rocket engine, it has stringent requirement on the braze process. Poor braze quality or damage to the parent materials limits the nozzle service life. The objective of this study was to develop a laser brazing process to provide quality, localized braze joints without adverse affect on the parent materials. Gold (Au-Cu-Ni-Pd-Mn) based high temperature braze alloys were used in both powder and wire form. Thin section iron base superalloy A286 tube was used as substrate materials. Different Laser Systems including CO2 (10.6 micrometers, 1kW), ND:YAG (1.06 micrometers, 4kW). and direct diode laser (808nm. 150W) were investigated for brazing process. The laser process variables including wavelength. laser power, travel speed and angle of inclination were optimized according to bead geometry and braze alloy wetting at minimum heat input level, The properties of laser brazing were compared to that of furnace brazing. Microhardness profiles were used for braze joint property comparison between laser and furnace brazing. The cooling rate of laser brazing was compared to furnace brazing based on secondary dendritic arm spacing, Both optical and Scanning Electron Microscope (SEM) were used to evaluate the microstructures of

  19. Effects of thermal cycling and thermal aging on the hermeticity and strength of silver-copper oxide air-brazed seals

    NASA Astrophysics Data System (ADS)

    Scott Weil, K.; Coyle, Christopher A.; Darsell, Jens T.; Xia, Gordon G.; Hardy, John S.

    Thermal cycle and exposure tests were conducted on ceramic-to-metal joints prepared by a new sealing technique. Known as reactive air brazing, this joining method is currently being considered for use in sealing various high-temperature solid-state electrochemical devices, including planar solid oxide fuel cells (pSOFC). In order to simulate a typical pSOFC application, test specimens were prepared by joining ceramic anode/electrolyte bilayers to metal washers, of the same composition as the common frame materials employed in pSOFC stacks, using a filler metal composed of 4 mol% CuO in silver. The brazed samples were exposure tested at 750 °C for 200, 400, and 800 h in both simulated fuel and air environments and thermally cycled at rapid rate (75 °C min -1) between room temperature and 750 °C for as many as 50 cycles. Subsequent joint strength testing and microstructural analysis indicated that the samples exposure tested in air displayed little degradation with respect to strength, hermeticity, or microstructure out to 800 h of exposure. Those tested in fuel showed no change in rupture strength or loss in hermeticity after 800 h of high-temperature exposure, but did undergo microstructural change due to the dissolution of hydrogen into the silver-based braze material. Air-brazed specimens subjected to rapid thermal cycling exhibited no loss in joint strength or hermeticity, but displayed initial signs of seal delamination along the braze-electrolyte interface after 50 cycles.

  20. Improved brazing technique

    DOEpatents

    Harast, D.G.

    1984-01-27

    A method of brazing comprises sand blasting the surfaces of the components to be brazed with particles of a brazing material to clean the surfaces and to embed brazing material in the surfaces, applying the brazing material to the surfaces, and heating the brazing material to form a brazement between the components.

  1. BRAZING ALLOYS

    DOEpatents

    Donnelly, R.G.; Gilliland, R.G.; Slaughter, G.M.

    1963-02-26

    A brazing alloy which, in the molten state, is characterized by excellent wettability and flowability, said alloy being capable of forming a corrosion resistant brazed joint wherein at least one component of said joint is graphite and the other component is a corrosion resistant refractory metal, said alloy consisting essentially of 20 to 50 per cent by weight of gold, 20 to 50 per cent by weight of nickel, and 15 to 45 per cent by weight of molybdenum. (AEC)

  2. Trends in wetting behavior for Ag–CuO braze alloys on Ba0.5Sr0.5Co0.8Fe0.2O(3-δ) at elevated temperatures in air

    SciTech Connect

    Joshi, Vineet V.; Meier, Alan; Darsell, Jens; Weil, K. Scott; Bowden, Mark

    2013-06-21

    Ba0.5Sr0.5Co0.80.2O(3-δ)(BSCF) is a potential oxygen separation membrane material for advanced coal based power plants. For this application, BSCF must be joined to a metal. In the current study, Ag-CuO, a reactive air brazing (RAB) alloy was evaluated for brazing BSCF. In-situ contact angle tests were performed on BSCF using Ag-CuO binary mixtures at 950 and 1000°C and the interfacial microstructures were evaluated. Wetting contact angles (θ< 90°) were obtained at short times at 950°C and the contact angles remained constant at 1000°C for 1, 2 and 8 mol% CuO contents. Microstructural analysis revealed the dissolution of copper oxide into the BSCF matrix to form copper-cobalt-oxygen rich dissolution products along the BSCF grain boundary. The formation of a thick interfacial reaction product layer and ridging at the sessile drop triple point indicate that the reaction kinetics are very rapid and that it will require careful process control to obtain the desired thin but continuous interfacial product layer.

  3. Reversible brazing process

    DOEpatents

    Pierce, Jim D.; Stephens, John J.; Walker, Charles A.

    1999-01-01

    A method of reversibly brazing surfaces together. An interface is affixed to each surface. The interfaces can be affixed by processes such as mechanical joining, welding, or brazing. The two interfaces are then brazed together using a brazing process that does not defeat the surface to interface joint. Interfaces of materials such as Ni-200 can be affixed to metallic surfaces by welding or by brazing with a first braze alloy. The Ni-200 interfaces can then be brazed together using a second braze alloy. The second braze alloy can be chosen so that it minimally alters the properties of the interfaces to allow multiple braze, heat and disassemble, rebraze cycles.

  4. Effects of CuO Content on the Wetting Behavior and Mechanical Properties of a Ag-CuO Braze for Ceramic Joining

    SciTech Connect

    Kim, Jin Yong Y.; Hardy, John S.; Weil, K. Scott

    2005-09-01

    A new silver-based joining technique referred to as reactive air brazing (RAB) has been recently developed for joining high temperature structural ceramic components of the type used in gas turbines, combustion engines, heat exchangers, and burners. It was found that additions of CuO to silver exhibit a tremendous effect on both the wettability and joint strength characteristics of the subsequent braze relative to polycrystalline alumina substrates. The effect is particularly significant at low CuO content, with substantial improvements in wetting observed in the 1 – 8 mol% range. The corresponding strength of the brazed polycrystalline alumina joints appears to be maximized at a copper oxide content of 8 mol%, with a maximum room temperature flexural strength approaching that of monolithic alumina. While further increases in oxide content lead to improved wetting on polycrystalline alumina, the effect on joint strength is deleterious. It appears that the formation of a continuous brittle copper-based oxide layer along the interface between the braze and alumina faying surface is responsible for the poor mechanical behavior observed in joints fabricated with higher CuO content brazes.

  5. The Effect of Palladium Additions on the Solidus/Liquidus Temperatures and Wetting Properties of Ag-CuO Based Air Brazes

    SciTech Connect

    Darsell, Jens T.; Weil, K. Scott

    2007-05-16

    As a means of increasing the use temperature of ceramic-ceramic and ceramic-metal air brazes, palladium was investigated as possible ternary addition to the currently employed silver - copper oxide system. The silver component was directly substituted with palladium to form the following series of alloys: (100-y)[(100-z)Pd - (z)Ag] - (y)CuOx where y = 0 - 34 mol% CuOx, z = 50 - 100 mol% silver, and x = 0, 0.5, and 1, denoting copper metal, Cu2O, or CuO. From differential scanning calorimetry, it was determined that the addition of palladium causes an increase in the solidus and liquidus temperatures of the resulting Pd-Ag-CuO brazes. In general, the liquidus was found to increase by approximately 220°C for the (100-y)(25Pd - 75Ag) - (z)CuOx filler metal compositions relative to comparable Ag-CuOx alloys. Likewise, the solidus was found to increase for these alloys, respectively by 185°C and 60°C, respectively for CuOx contents of y = 0 - 1mol% and 4 - 10 mol%. For the (100-y)(50Pd - 50Ag) - (y)CuOx alloys, the solidus increased between 280 - 390°C over a copper oxide compositional range of x = 0 to 8 mol%. It was determined from sessile drop experiments conducted on alumina substrates that in all cases the palladium causes an increase in the wetting angle relative to the corresponding binary braze. Alloy compositions of (100-y)(25Pd - 75Ag) - (y)CuOx displayed increased wetting angles of 5-20° relative to comparable binary compositions. (100-y)(50Pd - 50Ag) - (y)CuOx alloys exhibited an increase in contact angle of 10-60° and compositions containing less than 10 mol% CuOx were not able to wet the substrate. Scanning electron microscopy indicated that the microstructure of the braze consists of discrete CuOx precipitates in an alloyed silver-palladium matrix. In both the binary and ternary filler metal formulations, a reaction layer consisting of CuAlO2 was observed along the interface with the alumina substrate. This reaction product appears to be beneficial

  6. BRAZE BONDING OF COLUMBIUM

    DOEpatents

    Heestand, R.L.; Picklesimer, M.L.

    1962-07-31

    A method of brazing niobium parts together is described. The surfaces of the parts to be brazed together are placed in abutting relationship with a brazing alloy disposed adjacent. The alloy consists essentially of, by weight, 12 to 25% niobium, 0.5 to 5% molybdenum, and the balance zirconium, The alloy is heated to at least its melting point to braze the parts together. The brazed joint is then cooled. The heating, melting and cooling take place in an inert atmosphere. (AEC)

  7. The Effect of Composition on the Wetting Behavior and Joint Strength of the Ag-CuO Reactive Air Braze

    SciTech Connect

    Weil, K. Scott; Coyle, Christopher A.; Kim, Jin Yong Y.; Hardy, John S.

    2003-05-15

    One of the challenges in manufacturing solid-state electrochemical devices is in joining the ceramic and metallic components such that the resulting joint is rugged, hermetic, and stable under continuous high temperature operation in an oxidizing atmosphere. A well proven method of joining dissimilar materials is by brazing. Unfortunately many of the commercially available ceramic-to-metal braze alloys exhibit oxidation behavior which is unacceptable for potential use in a high temperature electrochemical device. An alternative braze alloy composition designed for oxidation resistance has been developed to join ferritic stainless steel to a variety of electrochemically active ceramic membranes including YSZ, nickel oxide, and mixed conducting perovskite oxides. The results of this study to date will be discussed.

  8. REACTIVE AIR BRAZING OF LSCoF AND ALUMINA WITH Ag-V2O5 ALLOYS FOR SOFC APPLICATIONS

    SciTech Connect

    Zink, Nathan M.; Meier, Alan; Weil, K. Scott; Hardy, John S.

    2005-05-31

    The feasibility of brazing LSCoF ceramic substrates with silver-vanadium pentoxide (Ag-V{sub 2}O{sub 5}) alloys for use in solid oxide fuel cell (SOFC) applications was studied. Preliminary testing was also performed on the Ag-V{sub 2}O{sub 5}/alumina (Al{sub 2}O{sub 3}) system in order to determine the feasibility of brazing LSCoF to Fecralloy{trademark} with an Ag-V{sub 2}O{sub 5} alloy since an Al{sub 2}O{sub 3} layer is formed on the Fecralloy surface prior to brazing. Sessile drop tests were performed using liquid Ag with 1 to 20 weight percent (wt%) V{sub 2}O{sub 5} additions at either 1000 or 1100 C. Similar to previous results for Ag-CuO alloys, additions of V{sub 2}O{sub 5} resulted in a decrease in the apparent contact angle and a transition from non-wetting to wetting behavior for both substrate materials. Mechanical test samples were fabricated by brazing using Ag with 1, 2 and 5 wt% V{sub 2}O{sub 5} additions. For the Ag-V{sub 2}O{sub 5}/LSCoF samples, the joint fracture strengths ranged from 10 to 30% of the monolithic LSCoF fracture strength ({sigma}{sub joint} = 26 {+-} 9 MPa versus {sigma}{sub LSCoF} = 151 {+-} 24 MPa). An SEM/EDS microstructural analysis of the brazed cross-sections indicated the formation of a vanadium rich reaction product at the braze/LSCoF interface. It is hypothesized that the reaction product provided improved chemical bonding at the interface that resulted in the modest joint strengths. In the the Ag-V{sub 2}O{sub 5}/Al{sub 2}O{sub 3} system, although the contact angles were less than 90{sup o}, all of the brazed Al{sub 2}O{sub 3} samples failed to bond during brazing or fractured during sample preparation. These poor adhesion results indicate potential problems with the Fecralloy{trademark}/Ag-V{sub 2}O{sub 5}/LSCoF system.

  9. Fluxless aluminum brazing

    DOEpatents

    Werner, W.J.

    1974-01-01

    This invention relates to a fluxless brazing alloy for use in forming brazed composites made from members of aluminum and its alloys. The brazing alloy consists of 35-55% Al, 10--20% Si, 25-60% Ge; 65-88% Al, 2-20% Si, 2--18% In; 65--80% Al, 15-- 25% Si, 5- 15% Y. (0fficial Gazette)

  10. Alumina barrier for vacuum brazing

    NASA Technical Reports Server (NTRS)

    Beuyukian, C. S.

    1980-01-01

    Heating platens of vacuum-brazing press will not stick to workpiece if aluminum oxide "paper" is interposed. Paper does not disintegrate in press, will not contaminate braze alloy, and helps form smoothly contoured, regular fillet at brazed edges.

  11. Novel interaction of Rab13 and Rab8 with endospanins☆

    PubMed Central

    Hirvonen, Mirkka J.; Büki, Kálmán G.; Sun, Yi; Mulari, Mika T.K.; Härkönen, Pirkko L.; Väänänen, Kalervo H.

    2013-01-01

    Rab GTPases regulate vesicular traffic in eukaryotic cells by cycling between the active GTP-bound and inactive GDP-bound states. Their functions are modulated by the diverse selection of effector proteins that bind to specific Rabs in their activated state. We previously described the expression of Rab13 in bone cells. To search for novel Rab13 interaction partners, we screened a newborn rat bone marrow cDNA library for Rab13 effectors with a bacterial two-hybrid system. We found that Rab13 binds to the C-terminus of Endospanin-2, a small transmembrane protein. In addition to Rab13 also Rab8 bound to Endospanin-2, while no binding of Rab7, Rab10, Rab11 or Rab32 was observed. Rab13 and Rab8 also interacted with Endospanin-1, a close homolog of Endospanin-2. Rab13 and Endospanin-2 colocalised in perinuclear vesicular structures in Cos1 cells suggesting direct binding also in vivo. Endospanin-2 is implicated in the regulation of the cell surface growth hormone receptor (GHR), but the inhibition of Rab13 expression did not affect GHR cell surface expression. This suggests that the Rab13–Endospanin-2 interaction may have functions other than GHR regulation. In conclusion, we have identified a novel interaction for Rab13 and Rab8 with Endospanin-2 and Endospanin-1. The role of this interaction in cell physiology, however, remains to be elucidated. PMID:23772379

  12. Interaction behaviors at the interface between liquid Al-Si and solid Ti-6Al-4V in ultrasonic-assisted brazing in air.

    PubMed

    Chen, Xiaoguang; Yan, Jiuchun; Gao, Fei; Wei, Jinghui; Xu, Zhiwu; Fan, Guohua

    2013-01-01

    Power ultrasonic vibration (20 kHz, 6 μm) was applied to assist the interaction between a liquid Al-Si alloy and solid Ti-6Al-4V substrate in air. The interaction behaviors, including breakage of the oxide film on the Ti-6Al-4V surface, chemical dissolution of solid Ti-6Al-4V, and interfacial chemical reactions, were investigated. Experimental results showed that numerous 2-20 μm diameter-sized pits formed on the Ti-6Al-4V surface. Propagation of ultrasonic waves in the liquid Al-Si alloy resulted in ultrasonic cavitation. When this cavitation occurred at or near the liquid/solid interface, many complex effects were generated at the small zones during the bubble implosion, including micro-jets, hot spots, and acoustic streaming. The breakage behavior of oxide films on the solid Ti-6Al-4V substrate, excessive chemical dissolution of solid Ti-6Al-4V into liquid Al-Si, abnormal interfacial chemical reactions at the interface, and phase transformation between the intermetallic compounds could be wholly ascribed to these ultrasonic effects. An effective bond between Al-Si and Ti-6Al-4V can be produced by ultrasonic-assisted brazing in air.

  13. Brazing Dissimilar Metals

    NASA Technical Reports Server (NTRS)

    Krotz, Phillip D.; Davis, William M.; Wisner, Daniel L.

    1996-01-01

    Brazing effective technique for joining ordinary structural metals to brittle, low-thermal-expansion refractory metals. Specifically, brazing process established for joining copper or nickel flanges to ends of vacuum-plasma-sprayed tungsten tubes and for joining stainless-steel flanges to ends of tubes made of alloy of molybdenum with 40 percent of rhenium.

  14. METHOD OF BRAZING

    DOEpatents

    Patriarca, P.; Slaughter, G.M.

    1962-11-27

    A method of joining metal surfaces is given. Surfaces having nickel or iron as the base metal are joined together with a brazing composition consisting of 80% nickel, 10% phosphorus, and 10% chromium. (AEC)

  15. Rab24 interacts with the Rab7/Rab interacting lysosomal protein complex to regulate endosomal degradation.

    PubMed

    Amaya, Celina; Militello, Rodrigo D; Calligaris, Sebastián D; Colombo, María I

    2016-11-01

    Endocytosis is a multistep process engaged in extracellular molecules internalization. Several proteins including the Rab GTPases family coordinate the endocytic pathway. The small GTPase Rab7 is present in late endosome (LE) compartments being a marker of endosome maturation. The Rab interacting lysosomal protein (RILP) is a downstream effector of Rab7 that recruits the functional dynein/dynactin motor complex to late compartments. In the present study, we have found Rab24 as a component of the endosome-lysosome degradative pathway. Rab24 is an atypical protein of the Rab GTPase family, which has been attributed a function in vesicle trafficking and autophagosome maturation. Using a model of transiently expressed proteins in K562 cells, we found that Rab24 co-localizes in vesicular structures labeled with Rab7 and LAMP1. Moreover, using a dominant negative mutant of Rab24 or a siRNA-Rab24 we showed that the distribution of Rab7 in vesicles depends on a functional Rab24 to allow DQ-BSA protein degradation. Additionally, by immunoprecipitation and pull down assays, we have demonstrated that Rab24 interacts with Rab7 and RILP. Interestingly, overexpression of the Vps41 subunit from the homotypic fusion and protein-sorting (HOPS) complex hampered the co-localization of Rab24 with RILP or with the lysosomal GTPase Arl8b, suggesting that Vps41 would affect the Rab24/RILP association. In summary, our data strongly support the hypothesis that Rab24 forms a complex with Rab7 and RILP on the membranes of late compartments. Our work provides new insights into the molecular function of Rab24 in the last steps of the endosomal degradative pathway.

  16. Rab6-interacting protein 1 links Rab6 and Rab11 function.

    PubMed

    Miserey-Lenkei, Stéphanie; Waharte, François; Boulet, Annick; Cuif, Marie-Hélène; Tenza, Danielle; El Marjou, Amed; Raposo, Graça; Salamero, Jean; Héliot, Laurent; Goud, Bruno; Monier, Solange

    2007-10-01

    Rab11 and Rab6 guanosine triphosphatases are associated with membranes of the recycling endosomes (REs) and Golgi complex, respectively. Evidence indicates that they sequentially regulate a retrograde transport pathway between these two compartments, suggesting the existence of proteins that must co-ordinate their functions. Here, we report the characterization of two isoforms of a protein, Rab6-interacting protein 1 (R6IP1), originally identified as a Rab6-binding protein. R6IP1 also binds to Rab11A in its GTP-bound conformation. In interphase cells, R6IP1 is targeted to the Golgi in a Rab6-dependent manner but can associate with Rab11-positive compartments when the level of Rab11A is increased within the cells. Fluorescence resonance energy transfer analysis using fluorescence lifetime imaging shows that the overexpression of R6IP1 promotes an interaction between Rab11A and Rab6 in living cells. Accordingly, the REs marked by Rab11 and transferrin receptor are depleted from the cell periphery and accumulate in the pericentriolar area. However, endosomal and Golgi membranes do not appear to fuse with each other. We also show that R6IP1 function is required during metaphase and cytokinesis, two mitotic steps in which a role of Rab6 and Rab11 has been previously documented. We propose that R6IP1 may couple Rab6 and Rab11 function throughout the cell cycle.

  17. Furnace brazing under partial vacuum

    NASA Technical Reports Server (NTRS)

    Mckown, R. D.

    1979-01-01

    Brazing furnace utilizing partial-vacuum technique reduces tooling requirements and produces better bond. Benefit in that partial vacuum helps to dissociate metal oxides that inhibit metal flow and eliminates heavy tooling required to hold parts together during brazing.

  18. Weld-brazing of titanium

    NASA Technical Reports Server (NTRS)

    Bales, T. T.; Royster, D. M.; Arnold, W. E., Jr.

    1974-01-01

    A joining process, designated weld-brazing, which combines resistance spotwelding and brazing has been developed at the NASA Langley Research Center. Resistance spot-welding is employed to position and align the parts and to establish a suitable faying surface gap for brazing; it contributes to the integrity of the joint. Brazing enhances the properties of the joint and reduces the stress concentrations normally associated with spotwelds. Ti-6Al-4V titanium alloy joints have been fabricated using 3003 aluminum braze both in a vacuum furnace and in a retort containing an inert gas environment.

  19. Active braze process

    SciTech Connect

    Levine, I.L.; Pike, R.A.

    1990-11-02

    Active metal bonding using Cusil (silver-copper) braze alloys is a well established method used at GE Neutron Devices (GEND) for bonding metal to metal, metal to ceramics, and ceramics to ceramics. However, there are many instances in which using a silver alloy for bonding is undesirable (e.g., in vacuum tube envelopes, or where sequential braze steps at different temperatures are required to complete an assembly). The Material and Processes Laboratory at GEND has discovered a new method of active brazing with non-silver alloys which has proved especially successful in ceramic-to-ceramic joints. This method has the added advantage of eliminating several steps which are required in conventional bonding techniques. 2 figs., 10 tabs.

  20. Characterisation of the Rab binding properties of Rab coupling protein (RCP) by site-directed mutagenesis.

    PubMed

    Lindsay, Andrew J; McCaffrey, Mary W

    2004-07-30

    Rab coupling protein (RCP) is a member of the Rab11-family of interacting proteins (Rab11-FIPs). Family members are characterised by their ability to interact with Rab11. This property is mediated by a conserved Rab binding domain (RBD) located at their carboxy-termini. Several Rab11-FIPs can also interact with other small GTPases. RCP interacts with Rab4 in addition to Rab11. To dissect out the individual properties of the Rab4 and Rab11 interactions with RCP, conserved amino acids within the RBD of RCP were mutated by site-directed mutagenesis. The effect of these mutations on Rab4 and Rab11 binding, and the intracellular localisation of RCP, was examined. Our results indicate that Rab11, rather than Rab4, mediates the intracellular localisation of RCP, and that the class I Rab11-FIPs compete for binding to Rab11.

  1. METHOD OF BRAZING BERYLLIUM

    DOEpatents

    Hanks, G.S.; Keil, R.W.

    1963-05-21

    A process is described for brazing beryllium metal parts by coating the beryllium with silver (65- 75 wt%)-aluminum alloy using a lithium fluoride (50 wt%)-lithium chloride flux, and heating the coated joint to a temperature of about 700 un. Concent 85% C for about 10 minutes. (AEC)

  2. Brazing of titanium by Cu-P brazing filler metals

    SciTech Connect

    Ariga, Tadashi; Matsu, Kotaro; Miyazawa, Yasuyuki

    1994-12-31

    Brazing of commercially pure titanium materials was made using the two types of the copper-phosphorus brazing filler metal foil. Two types of the brazing filler metal foils, BCuP-2 and -3 (AWS classification number), were used in this study. The chemical compositions of BCuP-2 and -3 were Cu-7.2P and Cu-6P-5Ag. Brazing was done in an argon gas atmosphere. After brazing, cutting, and polishing of the specimen, the mechanical properties of the specimen were estimated by shear strength test. And the cross-sectional microstructures at the brazed joint were observed by optical microscope and the elemental distributions at the brazed joint were analyzed by SENM and EPMA examinations. A sound brazed joint was obtained in this study. The maximum shear strength of the specimen was about 300 MPa in this study. According to observation of the cross-sectional microstructures, the reaction layer was formed at the base metal/brazed layer interface. The fracture during the shear test occurred in this reaction layer. And the phosphorus element was concentrated in this layer. Therefore, it appeared that the behavior of phosphorus element influenced the brazeability of these specimen.

  3. Brazing Refractory Metals Used In High-Temperature Nuclear Instrumentation

    SciTech Connect

    A. J. Palmer; C. J. Woolstenhulme

    2009-06-01

    As part of the U. S. Department of Energy (DOE) sponsored Next Generation Nuclear Project (NGNP) currently ongoing at Idaho National Laboratory (INL), the irradiation performance of candidate high-temperature gas reactor fuels and materials is being evaluated at INL’s Advanced Test Reactor (ATR). The design of the first Advanced Gas Reactor (AGR 1) experiment, currently being irradiated in the ATR, required development of special techniques for brazing niobium and molybdenum. Brazing is one technique used to join refractory metals to each other and to stainless steel alloys. Although brazing processes are well established, it is difficult to braze niobium, molybdenum, and most other refractory metals because they quickly develop adherent oxides when exposed to room-temperature air. Specialized techniques and methods were developed by INL to overcome these obstacles. This paper describes the techniques developed for removing these oxides, as well as the ASME Section IX-qualified braze procedures that were developed as part of the AGR-1 project. All brazes were made using an induction coil with an inert or reducing atmosphere at low pressure. Other parameters, such as filler metals, fluxes used, and general setup procedures, are also discussed.

  4. Vacuum brazing ceramics to metals

    SciTech Connect

    Mizuhara, H.

    1987-02-01

    Attention is given to the use in ceramic/metal joint brazing alloy of silver-copper composition that incorporates 2 percent Ti. This alloy allows one-step brazing, and wets superalloys and stainless steels without prior Ni plating of the substrate. Another alloy consisting of Ag-Cu-In-Ti has been developed which alloys at lower temperatures and allows step-brazing when used with Ag-Cu-Ti alloy. If the thermal expansion difference between metal and ceramic is large, brazing with a graded seal may be used; this minimizes joint stresses upon cooling to room temperature.

  5. Failure Assessment of Brazed Structures

    NASA Technical Reports Server (NTRS)

    Flom, Yuri

    2012-01-01

    Despite the great advances in analytical methods available to structural engineers, designers of brazed structures have great difficulties in addressing fundamental questions related to the loadcarrying capabilities of brazed assemblies. In this chapter we will review why such common engineering tools as Finite Element Analysis (FEA) as well as many well-established theories (Tresca, von Mises, Highest Principal Stress, etc) don't work well for the brazed joints. This chapter will show how the classic approach of using interaction equations and the less known Coulomb-Mohr failure criterion can be employed to estimate Margins of Safety (MS) in brazed joints.

  6. Rab coupling protein (RCP), a novel Rab4 and Rab11 effector protein.

    PubMed

    Lindsay, Andrew J; Hendrick, Alan G; Cantalupo, Giuseppina; Senic-Matuglia, Francesca; Goud, Bruno; Bucci, Cecilia; McCaffrey, Mary W

    2002-04-05

    Rab4 and Rab11 are small GTPases belonging to the Ras superfamily. They both function as regulators along the receptor recycling pathway. We have identified a novel 80-kDa protein that interacts specifically with the GTP-bound conformation of Rab4, and subsequent work has shown that it also interacts strongly with Rab11. We name this protein Rab coupling protein (RCP). RCP is predominantly membrane-bound and is expressed in all cell lines and tissues tested. It colocalizes with early endosomal markers including Rab4 and Rab11 as well as with the transferrin receptor. Overexpression of the carboxyl-terminal region of RCP, which contains the Rab4- and Rab11-interacting domain, results in a dramatic tubulation of the transferrin compartment. Furthermore, expression of this mutant causes a significant reduction in endosomal recycling without affecting ligand uptake or degradation in quantitative assays. RCP is a homologue of Rip11 and therefore belongs to the recently described Rab11-FIP family.

  7. Method for controlling brazing

    DOEpatents

    Hosking, F. Michael; Hall, Aaron C.; Givler, Richard C.; Walker, Charles A.

    2006-08-01

    A method for making a braze joint across a discontinuity in a work piece using alternating current. A filler metal is pre-placed at a location sufficiently close to the discontinuity such that, when an alternating current is applied across a work piece to heat the work piece and melt the filler metal, the filler metal is drawn into the discontinuity. The alternating current is maintained for a set residence time, generally less than 10 seconds and more particularly less than 3 seconds. The alternating current is then altered, generally by reducing the current and/or voltage such that the filler metal can solidify to form a braze joint of desired quality and thickness.

  8. Vacuum Brazing of Accelerator Components

    NASA Astrophysics Data System (ADS)

    Singh, Rajvir; Pant, K. K.; Lal, Shankar; Yadav, D. P.; Garg, S. R.; Raghuvanshi, V. K.; Mundra, G.

    2012-11-01

    Commonly used materials for accelerator components are those which are vacuum compatible and thermally conductive. Stainless steel, aluminum and copper are common among them. Stainless steel is a poor heat conductor and not very common in use where good thermal conductivity is required. Aluminum and copper and their alloys meet the above requirements and are frequently used for the above purpose. The accelerator components made of aluminum and its alloys using welding process have become a common practice now a days. It is mandatory to use copper and its other grades in RF devices required for accelerators. Beam line and Front End components of the accelerators are fabricated from stainless steel and OFHC copper. Fabrication of components made of copper using welding process is very difficult and in most of the cases it is impossible. Fabrication and joining in such cases is possible using brazing process especially under vacuum and inert gas atmosphere. Several accelerator components have been vacuum brazed for Indus projects at Raja Ramanna Centre for Advanced Technology (RRCAT), Indore using vacuum brazing facility available at RRCAT, Indore. This paper presents details regarding development of the above mentioned high value and strategic components/assemblies. It will include basics required for vacuum brazing, details of vacuum brazing facility, joint design, fixturing of the jobs, selection of filler alloys, optimization of brazing parameters so as to obtain high quality brazed joints, brief description of vacuum brazed accelerator components etc.

  9. Brazing in Space: The Next Frontier

    NASA Technical Reports Server (NTRS)

    Flom, Yury

    2005-01-01

    This viewgraph presentation provides information on the challenges facing mechanically joining objects in space, as well as past, present, and future techniques for brazing. The sections of the presentation include: 1) Why do we need to join components in space; 2) Why brazing? 3) History of brazing in space; 4) Electron beam vacuum brazing; 5) Current effort at GSFC; 6) Future work.

  10. Characterisation of Ga-coated and Ga-brazed aluminium

    SciTech Connect

    Ferchaud, E.; Christien, F.; Barnier, V.; Paillard, P.

    2012-05-15

    This work is devoted to the brazing of aluminium using liquid gallium. Gallium was deposited on aluminium samples at {approx} 50 Degree-Sign C using a liquid gallium 'polishing' technique. Brazing was undertaken for 30 min at 500 Degree-Sign C in air. EDS (Energy Dispersive X-ray Spectroscopy) and AES (Auger Electron Spectroscopy) characterisation of Ga-coated samples has shown that the Ga surface layer thickness is of ten (or a few tens of) nanometres. Furthermore, aluminium oxide layer (Al{sub 2}O{sub 3}) was shown to be 'descaled' during Ga deposition, which ensures good conditions for further brazing. Cross-section examination of Ga-coated samples shows that liquid gallium penetrates into the aluminium grain boundaries during deposition. The thickness of the grain boundary gallium film was measured using an original EDS technique and is found to be of a few tens of nanometres. The depth of gallium grain boundary penetration is about 300 {mu}m at the deposition temperature. The fracture stress of the brazed joints was measured from tensile tests and was determined to be 33 MPa. Cross-section examination of brazed joints shows that gallium has fully dissolved into the bulk and that the joint is really autogenous. - Highlights: Black-Right-Pointing-Pointer Aluminium can be brazed using liquid gallium deposited by a 'polishing' technique. Black-Right-Pointing-Pointer The aluminium oxide layer is 'descaled' during liquid Ga 'polishing' deposition. Black-Right-Pointing-Pointer EDS can be used for determination of surface and grain boundary Ga film thickness. Black-Right-Pointing-Pointer The surface and grain boundary Ga film thickness is of a few tens of nm. Black-Right-Pointing-Pointer Surface and grain boundary gallium dissolves in the bulk during brazing.

  11. Brazing graphite to graphite

    DOEpatents

    Peterson, George R.

    1976-01-01

    Graphite is joined to graphite by employing both fine molybdenum powder as the brazing material and an annealing step that together produce a virtually metal-free joint exhibiting properties similar to those found in the parent graphite. Molybdenum powder is placed between the faying surfaces of two graphite parts and melted to form molybdenum carbide. The joint area is thereafter subjected to an annealing operation which diffuses the carbide away from the joint and into the graphite parts. Graphite dissolved by the dispersed molybdenum carbide precipitates into the joint area, replacing the molybdenum carbide to provide a joint of virtually graphite.

  12. Development of brazing foils to join monocrystalline tungsten alloys with ODS-EUROFER steel

    NASA Astrophysics Data System (ADS)

    Kalin, B. A.; Fedotov, V. T.; Sevrjukov, O. N.; Kalashnikov, A. N.; Suchkov, A. N.; Moeslang, A.; Rohde, M.

    2007-08-01

    Results on rapidly solidified filler metals for brazing W with W and monocrystalline W with EUROFER steel (FS) are presented. Rapidly quenched powder-type filler metals based on Ti bal-V-Cr-Be were developed to braze polycrystalline W with monocrystalline W. In addition, Fe bal-Ta-Ge-Si-B-Pd alloys were developed to braze monocrystalline W with FS for helium gas cooled divertors and plasma-facing components. The W to FS brazed joints were fabricated under vacuum at 1150 °C, using a Ta spacer of 0.1 mm in thickness to account for the different thermal expansions. The monocrystalline tungsten as well as the related brazed joints withstood 30 cycles between 750 °C/20 min and air cooling/3-5 min.

  13. Thermally stable diamond brazing

    DOEpatents

    Radtke, Robert P.

    2009-02-10

    A cutting element and a method for forming a cutting element is described and shown. The cutting element includes a substrate, a TSP diamond layer, a metal interlayer between the substrate and the diamond layer, and a braze joint securing the diamond layer to the substrate. The thickness of the metal interlayer is determined according to a formula. The formula takes into account the thickness and modulus of elasticity of the metal interlayer and the thickness of the TSP diamond. This prevents the use of a too thin or too thick metal interlayer. A metal interlayer that is too thin is not capable of absorbing enough energy to prevent the TSP diamond from fracturing. A metal interlayer that is too thick may allow the TSP diamond to fracture by reason of bending stress. A coating may be provided between the TSP diamond layer and the metal interlayer. This coating serves as a thermal barrier and to control residual thermal stress.

  14. Brazeability of powder aluminum brazing filler metals with non-corrosive flux

    SciTech Connect

    Takemoto, T.; Matsunawa, A.; Ujie, T.

    1994-12-31

    Various brazed aluminum products, mainly heat exchangers, have been widely used especially in automobiles and electric power industries. They have been produced by using brazing sheet; however, recently the demand to braze the complicated shape is increasing, leading to the necessity for setting brazing filler metal at braze parts instead of using brazing sheet. Therefore, the present work aimed to investigated the brazeability of aluminum powder filler metal in nitrogen gas atmosphere using KAIF{sub 4}-K{sub 3}AIF{sub 6} system noncorrosive brazing flux. By considering the applicability of filler metal, brazing pastes were made of powder filler metal, flux and organic binder. AI-Si powder brazing filler metals were made by automization. T-type specimen was made by A3003 base metal with thickness of 2 mm. in the present experiment, fillet formabiltity, the percentage of the length of formed fillet against the length of vertical member wall at each side, was adopted to evaluate brazeability. The shape of the atomized powder depended on atomize atmosphere and atomizing gas. Sound fillet formation was achieved on the full length of both sides of the vertical member under the condition of appropriate surface treatment and sufficient flux content. Decrease in flux content gave partial fillet formation at the opposite side of the paste set side. Further decrease brought the partial fillet formation at the paste set side also. The paste made of air-atomized powder required more flux content to achieve 100% fillet formation at the opposite side. On the other hand, argon-atomized powders formed fillet in full length using paste with less flux content. powders sorted to remove fine particles and powders with low oxygen content were found to be suitable for brazing filler metal powders, because they required less flux content to obtain 100% fillet formation under the same amount of paste.

  15. Vacuum Brazing of Beryllium Copper Components for the National Ignition Facility

    SciTech Connect

    Tyhurst, C.C.; Cunningham, M.A.

    2002-06-04

    A process for vacuum brazing beryllium copper anode assemblies was required for the Plasma Electrode Pockels Cell System, or PEPC, a component for the National Ignition Facility (NIF). Initial problems with the joint design and wettability of the beryllium copper drove some minor design changes. Brazing was facilitated by plating the joint surface of the beryllium copper rod with silver 0.0006 inch thick. Individual air sampling during processing and swipe tests of the furnace interior after brazing revealed no traceable levels of beryllium.

  16. Rab18 and a Rab18 GEF complex are required for normal ER structure

    PubMed Central

    Gerondopoulos, Andreas; Bastos, Ricardo Nunes; Yoshimura, Shin-ichiro; Anderson, Rachel; Carpanini, Sarah; Aligianis, Irene

    2014-01-01

    The ancestral Rab GTPase Rab18 and both subunits of the Rab3GAP complex are mutated in the human neurological and developmental disorder Warburg Micro syndrome. Here, we demonstrate that the Rab3GAP complex is a specific Rab18 guanine nucleotide exchange factor (GEF). The Rab3GAP complex localizes to the endoplasmic reticulum (ER) and is necessary for ER targeting of Rab18. It is also sufficient to promote membrane recruitment of Rab18. Disease-associated point mutations of conserved residues in either the Rab3GAP1 (T18P and E24V) or Rab3GAP2 (R426C) subunits result in loss of the Rab18 GEF and membrane-targeting activities. Supporting the view that Rab18 activity is important for ER structure, in the absence of either Rab3GAP subunit or Rab18 function, ER tubular networks marked by reticulon 4 were disrupted, and ER sheets defined by CLIMP-63 spread out into the cell periphery. Micro syndrome is therefore a disease characterized by direct loss of Rab18 function or loss of Rab18 activation at the ER by its GEF Rab3GAP. PMID:24891604

  17. Protective coating for salt-bath brazing

    NASA Technical Reports Server (NTRS)

    Francisco, A. C.; Gyorgak, C. A.

    1971-01-01

    Ceramic coating, consisting of graphite, enameler's clay, and algin binder, applied to materials prior to salt bath brazing facilitates brazing process and results in superior joints. Alternate coating materials and their various proportions are given.

  18. Induction brazing at the Kennedy Space Center

    NASA Technical Reports Server (NTRS)

    Clautice, W. E.

    1974-01-01

    A description of the joint design, materials, equipment, qualification testing, inspection methods, and applications involved in performing induction brazing on hyperbolic propellants tubing at Kennedy Space Center. Induction brazing is a form of brazing in which the energy is transmitted to the workpiece by electrical induction; the eddy currents generated in the metal produce heat by resistance losses. Since induction heating is fast and highly localized, undesirable heat effects are minimized and the resulting braze is of high quality.

  19. 32 CFR 202.10 - RAB adjournment and dissolution.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 2 2013-07-01 2013-07-01 false RAB adjournment and dissolution. 202.10 Section... dissolution. (a) RAB adjournment—(1) Requirements for RAB adjournment. An Installation Commander may adjourn a... Commander decides to adjourn the RAB. (b) RAB dissolution—(1) Requirements for RAB dissolution....

  20. 32 CFR 202.10 - RAB adjournment and dissolution.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 2 2012-07-01 2012-07-01 false RAB adjournment and dissolution. 202.10 Section... dissolution. (a) RAB adjournment—(1) Requirements for RAB adjournment. An Installation Commander may adjourn a... Commander decides to adjourn the RAB. (b) RAB dissolution—(1) Requirements for RAB dissolution....

  1. 32 CFR 202.10 - RAB adjournment and dissolution.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 2 2011-07-01 2011-07-01 false RAB adjournment and dissolution. 202.10 Section... dissolution. (a) RAB adjournment—(1) Requirements for RAB adjournment. An Installation Commander may adjourn a... Commander decides to adjourn the RAB. (b) RAB dissolution—(1) Requirements for RAB dissolution....

  2. 32 CFR 202.10 - RAB adjournment and dissolution.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 2 2014-07-01 2014-07-01 false RAB adjournment and dissolution. 202.10 Section... dissolution. (a) RAB adjournment—(1) Requirements for RAB adjournment. An Installation Commander may adjourn a... Commander decides to adjourn the RAB. (b) RAB dissolution—(1) Requirements for RAB dissolution....

  3. 32 CFR 202.10 - RAB adjournment and dissolution.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 2 2010-07-01 2010-07-01 false RAB adjournment and dissolution. 202.10 Section... dissolution. (a) RAB adjournment—(1) Requirements for RAB adjournment. An Installation Commander may adjourn a... Commander decides to adjourn the RAB. (b) RAB dissolution—(1) Requirements for RAB dissolution....

  4. Distinct actions of Rab3 and Rab27 GTPases on late stages of exocytosis of insulin.

    PubMed

    Cazares, Victor A; Subramani, Arasakumar; Saldate, Johnny J; Hoerauf, Widmann; Stuenkel, Edward L

    2014-09-01

    Rab GTPases associated with insulin-containing secretory granules (SGs) are key in targeting, docking and assembly of molecular complexes governing pancreatic β-cell exocytosis. Four Rab3 isoforms along with Rab27A are associated with insulin granules, yet elucidation of the distinct roles of these Rab families on exocytosis remains unclear. To define specific actions of these Rab families we employ Rab3GAP and/or EPI64A GTPase-activating protein overexpression in β-cells from wild-type or Ashen mice to selectively transit the entire Rab3 family or Rab27A to a GDP-bound state. Ashen mice carry a spontaneous mutation that eliminates Rab27A expression. Using membrane capacitance measurements we find that GTP/GDP nucleotide cycling of Rab27A is essential for generation of the functionally defined immediately releasable pool (IRP) and central to regulating the size of the readily releasable pool (RRP). By comparison, nucleotide cycling of Rab3 GTPases, but not of Rab27A, is essential for a kinetically rapid filling of the RRP with SGs. Aside from these distinct functions, Rab3 and Rab27A GTPases demonstrate considerable functional overlap in building the readily releasable granule pool. Hence, while Rab3 and Rab27A cooperate to generate release-ready SGs in β-cells, they also direct unique kinetic and functional properties of the exocytotic pathway.

  5. Restoration Advisory Board (RAB) Implementation Guidelines

    EPA Pesticide Factsheets

    This document provides recommended procedures for establishing and operating Restoration Advisory Boards (RABs). It is intended as a resource for installation, EPA, and State personnel and citizens who participate in RABs.

  6. Silver-hafnium braze alloy

    DOEpatents

    Stephens, Jr., John J.; Hosking, F. Michael; Yost, Frederick G.

    2003-12-16

    A binary allow braze composition has been prepared and used in a bonded article of ceramic-ceramic and ceramic-metal materials. The braze composition comprises greater than approximately 95 wt % silver, greater than approximately 2 wt % hafnium and less than approximately 4.1 wt % hafnium, and less than approximately 0.2 wt % trace elements. The binary braze alloy is used to join a ceramic material to another ceramic material or a ceramic material, such as alumina, quartz, aluminum nitride, silicon nitride, silicon carbide, and mullite, to a metal material, such as iron-based metals, cobalt-based metals, nickel-based metals, molybdenum-based metals, tungsten-based metals, niobium-based metals, and tantalum-based metals. A hermetic bonded article is obtained with a strength greater than 10,000 psi.

  7. Weld-brazing - a new joining process. [combination resistance spot welding and brazing of titanium alloys

    NASA Technical Reports Server (NTRS)

    Bales, T. T.; Royster, D. M.; Arnold, W. E., Jr.

    1972-01-01

    A joining process designated weld brazing which combines resistance spot welding and brazing has been developed. Resistance spot welding is used to position and align the parts as well as to establish a suitable faying surface gap for brazing. Fabrication is then completed by capillary flow of the braze alloy into the joint. The process has been used successfully to fabricate Ti-6Al-4V titanium alloy joints using 3003 aluminum braze alloy. Test results obtained on single overlap and hat-stiffened structural specimens show that weld brazed joints are superior in tensile shear, stress rupture, fatigue, and buckling than joint fabricated by spotwelding or brazing. Another attractive feature of the process is that the brazed joints is hermetically sealed by the braze material.

  8. Strength testing of Ti-vapor-coated silicon nitride braze joints

    SciTech Connect

    Santella, M.L.

    1994-09-01

    Sintered silicon nitride was vacuum brazed to itself at 1130{degree}C with a Au-25Ni-25Pd wt % filler metal. Wetting was obtained by coating the Si{sub 3}N{sub 4} surfaces with titanium prior to brazing by electron beam evaporation. The brazed joints were virtually free of porosity. Metallographic analysis showed that Ti reacted with the Si{sub 3}N{sub 4}, to form a TiN reaction layer during brazing. Small amounts of Si and Ti dissolved in the filler metal layers but they did not appear to influence the mechanical properties of the braze layer. Flexure bars were made from the brazed coupons and tested at room temperature, 600C, 700C, and 800C in air. At 700C and below, fracture of the test bars occurred in the Si{sub 3}N{sub 4}, either near the brazed surfaces or at some distance into the monolithic material. The measured strength of joint specimens decreased slightly with increasing test temperature, and generally exceeded the intrinsic braze filler metal strength in this temperature range. It was also found that lapping the Si{sub 3}N{sub 4}, prior to Ti coating reduced the number of near-surface flaws and produced joints with higher average strength and lower scatter than those left in a ground condition. Specimens tested at 800C had very low strengths, and this behavior was related to the microstructure at the brazed Si{sub 3}N{sub 4} surfaces.

  9. Field installed brazed thermocouple feedthroughs for high vacuum experiments

    NASA Astrophysics Data System (ADS)

    Anderson, P. M.; Messick, C.

    1983-12-01

    In order to reduce the occurrence of vacuum leaks and to increase the availability of the DIII vacuum vessel for experimental operation, effort was applied to developing a vacuum-tight brazed feedthrough system for sheathed thermocouples, stainless steel sheathed conductor cables and tubes for cooling fluids. This brazed technique is a replacement for elastomer O ring sealed feedthroughs that have proven vulnerable to leaks caused by thermal cycling, etc. To date, about 200 feedthroughs were used. Up to 91 were grouped on a single conflat flange mounted in a bulkhead connector configuration which facilitates installation and removal. Investigation was required to select a suitable braze alloy, flux and installation procedure. Braze alloy selection was challenging since the alloy was required to have: (1) melting temperature in excess of the 250 C (482 F) bakeout temperature; (2) no high vapor pressure elements; (3) good wetting properties when used in air with acceptable flux; and (4) good wettability to 300 series stainless steel and Inconel.

  10. Surface preparation for high purity alumina ceramics enabling direct brazing in hydrogen atmospheres

    DOEpatents

    Cadden, Charles H.; Yang, Nancy Yuan Chi; Hosking, Floyd M.

    2001-01-01

    The present invention relates to a method for preparing the surface of a high purity alumina ceramic or sapphire specimen that enables direct brazing in a hydrogen atmosphere using an active braze alloy. The present invention also relates to a method for directly brazing a high purity alumina ceramic or sapphire specimen to a ceramic or metal member using this method of surface preparation, and to articles produced by this brazing method. The presence of silicon, in the form of a SiO.sub.2 -containing surface layer, can more than double the tensile bond strength in alumina ceramic joints brazed in a hydrogen atmosphere using an active Au-16Ni-0.75 Mo-1.75V filler metal. A thin silicon coating applied by PVD processing can, after air firing, produce a semi-continuous coverage of the alumina surface with a SiO.sub.2 film. Room temperature tensile strength was found to be proportional to the fraction of air fired surface covered by silicon-containing films. Similarly, the ratio of substrate fracture versus interface separation was also related to the amount of surface silicon present prior to brazing. This process can replace the need to perform a "moly-manganese" metallization step.

  11. Structure-Function Analyses of the Interactions between Rab11 and Rab14 Small GTPases with Their Shared Effector Rab Coupling Protein (RCP).

    PubMed

    Lall, Patrick; Lindsay, Andrew J; Hanscom, Sara; Kecman, Tea; Taglauer, Elizabeth S; McVeigh, Una M; Franklin, Edward; McCaffrey, Mary W; Khan, Amir R

    2015-07-24

    Rab GTPases recruit effector proteins, via their GTP-dependent switch regions, to distinct subcellular compartments. Rab11 and Rab25 are closely related small GTPases that bind to common effectors termed the Rab11 family of interacting proteins (FIPs). The FIPs are organized into two subclasses (class I and class II) based on sequence and domain organization, and both subclasses contain a highly conserved Rab-binding domain at their C termini. Yeast two-hybrid and biochemical studies have revealed that the more distantly related Rab14 also interacts with class I FIPs. Here, we perform detailed structural, thermodynamic, and cellular analyses of the interactions between Rab14 and one of the class I FIPs, the Rab-coupling protein (RCP), to clarify the molecular aspects of the interaction. We find that Rab14 indeed binds to RCP, albeit with reduced affinity relative to conventional Rab11-FIP and Rab25-FIP complexes. However, in vivo, Rab11 recruits RCP onto biological membranes. Furthermore, biophysical analyses reveal a noncanonical 1:2 stoichiometry between Rab14-RCP in dilute solutions, in contrast to Rab11/25 complexes. The structure of Rab14-RCP reveals that Rab14 interacts with the canonical Rab-binding domain and also provides insight into the unusual properties of the complex. Finally, we show that both the Rab coupling protein and Rab14 function in neuritogenesis.

  12. Structures of Drosophila melanogaster Rab2 and Rab3 bound to GMPPNP

    PubMed Central

    Lardong, Jennifer A.; Driller, Jan H.; Depner, Harald; Weise, Christoph; Petzoldt, Astrid; Wahl, Markus C.; Sigrist, Stephan J.; Loll, Bernhard

    2015-01-01

    Rab GTPases belong to the large family of Ras proteins. They act as key regulators of membrane organization and intracellular trafficking. Functionally, they act as switches. In the active GTP-bound form they can bind to effector proteins to facilitate the delivery of transport vesicles. Upon stimulation, the GTP is hydrolyzed and the Rab proteins undergo conformational changes in their switch regions. This study focuses on Rab2 and Rab3 from Drosophila melanogaster. Whereas Rab2 is involved in vesicle transport between the Golgi and the endoplasmatic reticulum, Rab3 is a key player in exocytosis, and in the synapse it is involved in the assembly of the presynaptic active zone. Here, high-resolution crystal structures of Rab2 and Rab3 in complex with GMPPNP and Mg2+ are presented. In the structure of Rab3 a modified cysteine residue is observed with an enigmatic electron density attached to its thiol function. PMID:25615965

  13. 31 Flavors of Drosophila Rab proteins

    SciTech Connect

    Zhang, Jun; Schulze, Karen L.; Hiesinger, P. Robin; Suyama, Kaye; Wang, Stream; Fish, Matthew; Acar, Melih; Hoskins, Roger A.; Bellen, HugoJ.; Scott, Matthew P.

    2007-04-03

    Rab proteins are small GTPases that play important roles intransport of vesicle cargo and recruitment, association of motor andother proteins with vesicles, and docking and fusion of vesicles atdefined locations. In vertebrates, more than 75 Rab genes have beenidentified, some of which have been intensively studied for their rolesin endosome and synaptic vesicle trafficking. Recent studies of thefunctions of certain Rab proteins have revealed specific roles inmediating developmental signal transduction. We have begun a systematicgenetic study of the 33 Rab genes in Drosophila. Most of the fly proteinsare clearly related to specific vertebrate proteins. We report here thecreation of a set of transgenic fly lines that allow spatially andtemporally regulated expression of Drosophila Rab proteins. We generatedfluorescent protein-tagged wild-type, dominant-negative, andconstitutively active forms of 31 Drosophila Rab proteins. We describeDrosophila Rab expression patterns during embryogenesis, the subcellularlocalization of some Rab proteins, and comparisons of the localization ofwild-type, dominant-negative, and constitutively active forms of selectedRab proteins. The high evolutionary conservation and low redundancy ofDrosophila Rab proteins make these transgenic lines a useful toolkit forinvestigating Rab functions in vivo.

  14. Diffusion brazing of nickel aluminides

    SciTech Connect

    Orel, S.V.; Parous, L.C.; Gale, W.F.

    1995-09-01

    NiAl is a promising candidate material for high-temperature applications. However, NiAl suffers from poor low-temperature ductility and toughness. Hence, suitable technologies are required for NiAl to Ni-based alloy joining. In view of the poor low-temperature ductility and strong alumina-forming tendency of NiAl, diffusion brazing seems to be the most suitable technology for joining NiAl to itself and to Ni-based alloys. This paper examines the diffusion brazing of NiAl to Ni using Ni-Si-B interlayers and draws comparisons with previous work by authors on NiAl/Ni-Si-B/NiAl and Ni/Ni-Si-B/Ni diffusion brazing. The progression of micro-structural development in the NiAl/Ni-Si-B/Ni joints is compared with that expected from standard models of the diffusion brazing process in which dissolution of the substrate material, isothermal solidification and solid-state homogenization occur sequentially.

  15. Brazing C-C composites to metals

    SciTech Connect

    Liu, J.Y.; Banerjee, P.; Chin, B.A.

    1994-12-31

    Carbon-carbon composites are attractive for use at high temperature because of their high strength, modulus, chemical stability and resistance to activation by radiation. In these applications, the C-C composite must be joined to itself and to metals. The research described in this paper has led to the invention of a new brazing filler metal from the Cu-Mu-Ti system and the development of a brazing process for joining CC composites to metals. The newly invented brazing filler metal compositions, with controllable melting points ranging from 800{emdash}920{degrees}C has excellent wettability on both C-C composites and metals (stainless steel, Nb, Mo, W, and Zr). Sound joints of C-C composite/metal were produced using the brazing filler metal and the specially developed brazing processes. Finite element analyses were used to predict the residual stress distribution in the brazed joints. Theoretical predictions were confirmed by interlayer experiments. The brazed joints were studied using optical and scanning electron microscopy (SEM) to examine the microstructure and fractured brazed joints. The results showed 100% bonding was obtained using the developed braze metal and brazing procedure.

  16. Simulation based analysis of laser beam brazing

    NASA Astrophysics Data System (ADS)

    Dobler, Michael; Wiethop, Philipp; Schmid, Daniel; Schmidt, Michael

    2016-03-01

    Laser beam brazing is a well-established joining technology in car body manufacturing with main applications in the joining of divided tailgates and the joining of roof and side panels. A key advantage of laser brazed joints is the seam's visual quality which satisfies highest requirements. However, the laser beam brazing process is very complex and process dynamics are only partially understood. In order to gain deeper knowledge of the laser beam brazing process, to determine optimal process parameters and to test process variants, a transient three-dimensional simulation model of laser beam brazing is developed. This model takes into account energy input, heat transfer as well as fluid and wetting dynamics that lead to the formation of the brazing seam. A validation of the simulation model is performed by metallographic analysis and thermocouple measurements for different parameter sets of the brazing process. These results show that the multi-physical simulation model not only can be used to gain insight into the laser brazing process but also offers the possibility of process optimization in industrial applications. The model's capabilities in determining optimal process parameters are exemplarily shown for the laser power. Small deviations in the energy input can affect the brazing results significantly. Therefore, the simulation model is used to analyze the effect of the lateral laser beam position on the energy input and the resulting brazing seam.

  17. Fluxless Brazing of Large Structural Panels

    NASA Technical Reports Server (NTRS)

    Beuyukian, C. S.

    1982-01-01

    Fluxless brazing is used in fabricating aluminum structural panels that withstand high internal pressure. Aluminum sheet of structural thickness with 4045 aluminum/silicon-braze-alloy cladding is brazed to corrugated "fin stock" having channels 0.001 inch (0.03mm) high by same width. Process is carried out in an inert (argon) atmosphere in a retort furnace. Filler bars are used in some channels to prevent fin stock from collapsing as pressure is applied.

  18. Brazing of beryllium for structural applications

    NASA Technical Reports Server (NTRS)

    Vogan, J. W.

    1972-01-01

    Progress made in fabricating a beryllium compression tube structure and a stiffened beryllium panel. The compression tube was 7.6cm in diameter and 30.5cm long with titanium end fittings. The panel was 203cm long and stiffened with longitudinal stringers. Both units were assembled by brazing with BAg-18 braze alloy. The detail parts were fabricated by hot forming 0.305cm beryllium sheet and the brazing parameters established.

  19. Susceptor heating device for electron beam brazing

    DOEpatents

    Antieau, Susan M.; Johnson, Robert G. R.

    1999-01-01

    A brazing device and method are provided which locally apply a controlled amount of heat to a selected area, within a vacuum. The device brazes two components together with a brazing metal. A susceptor plate is placed in thermal contact with one of the components. A serrated pedestal supports the susceptor plate. When the pedestal and susceptor plate are in place, an electron gun irradiates an electron beam at the susceptor plate such that the susceptor plate is sufficiently heated to transfer heat through the one component and melt the brazing metal.

  20. Intelligent control of vacuum aluminum brazing

    SciTech Connect

    Zhong, G.; Pai, D.M.; Badgley, S.

    1995-06-01

    Vacuum brazing is a versatile modern day metal joining method. It usually includes vacuum producing, heating and residual gas analyzing systems. When used to join aluminum parts, the quality of the brazed joint is highly dependent on the residual gases in the vacuum, especially the residual oxygen and water vapor in the vacuum chamber which affect the formation of oxides. These vacuum environment brazing contaminants are reduced during the out-gassing processes of degassing and desorption during heating under vacuum. Exceeding the boundary limits of these contaminants causes unacceptable flow of braze filler in the joints during the brazing process. Identification of the quality control boundary for vacuum brazing of aluminum alloy makes computer control of vacuum brazing quality for aluminum alloys possible. The present study utilizes a residual gas analyzer (RGA) to monitor the partial pressures of residual gases during brazing. These data are transported to computer through the RS-232 interface port on the RGA, and used in real time to monitor the brazing quality by an algorithm based on the quality control boundary. If the quality is bad, the computer will send a ``hold`` signal to the heating system via another Rs-232 port until the environment is reestablished within the acceptable boundary.

  1. Thermal resilient multiple jaw braze fixture

    DOEpatents

    Ney, Robert; Perrone, Alex J.

    1995-07-11

    A braze fixture has side walls forming a cavity with an opening to receive a stack of parts to be brazed. Sidewalls of the housing have a plurality of bearing receiving openings into which bearing rods or jaws are inserted to align the stacked elements of the workpiece. The housing can also have view ports to allow a visual check of the alignment. Straps or wires around the fixture are selected to have thermal characteristics similar to the thermal characteristics of the workpiece undergoing brazing. The straps or wires make physical contact with the bearing rods thereby causing bearing rods to maintain the workpiece in proper alignment throughout the entire brazing cycle.

  2. Brazing Inconel 625 Using the Copper Foil

    NASA Astrophysics Data System (ADS)

    Chen, Wen-Shiang; Wang, Cheng-Yen; Shiue, Ren-Kae

    2013-12-01

    Brazing Inconel 625 (IN-625) using the copper foil has been investigated in this research. The brazed joint is composed of nanosized CrNi3 precipitates and Cr/Mo/Nb/Ni quaternary compound in the Cu/Ni-rich matrix. The copper filler 50 μm in thickness is enough for the joint filling. However, the application of Cu foil 100 μm in thickness has little effect on the shear strength of the brazed joint. The specimen brazed at 1433 K (1160 °C) for 1800 seconds demonstrates the best shear strength of 470 MPa, and its fractograph is dominated by ductile dimple fracture with sliding marks. Decreasing the brazing temperature slightly decreases the shear strength of the brazed joint due to the presence of a few isolated solidification shrinkage voids smaller than 15 μm. Increasing the brazing temperature, especially for the specimen brazed at 1473 K (1200 °C), significantly deteriorates the shear strength of the joint below 260 MPa because of coalescence of isothermal solidification shrinkage voids in the joint. The Cu foil demonstrates potential in brazing IN-625 for industrial application.

  3. Drosophila Pkaap regulates Rab4/Rab11-dependent traffic and Rab11 exocytosis of innate immune cargo

    PubMed Central

    Sorvina, Alexandra; Shandala, Tetyana; Brooks, Douglas A.

    2016-01-01

    ABSTRACT The secretion of immune-mediators is a critical step in the host innate immune response to pathogen invasion, and Rab GTPases have an important role in the regulation of this process. Rab4/Rab11 recycling endosomes are involved in the sorting of immune-mediators into specialist Rab11 vesicles that can traffic this cargo to the plasma membrane; however, how this sequential delivery process is regulated has yet to be fully defined. Here, we report that Drosophila Pkaap, an orthologue of the human dual-specific A-kinase-anchoring protein 2 or D-AKAP2 (also called AKAP10), appeared to have a nucleotide-dependent localisation to Rab4 and Rab11 endosomes. RNAi silencing of pkaap altered Rab4/Rab11 recycling endosome morphology, suggesting that Pkaap functions in cargo sorting and delivery in the secretory pathway. The depletion of pkaap also had a direct effect on Rab11 vesicle exocytosis and the secretion of the antimicrobial peptide Drosomycin at the plasma membrane. We propose that Pkaap has a dual role in antimicrobial peptide traffic and exocytosis, making it an essential component for the secretion of inflammatory mediators and the defence of the host against pathogens. PMID:27190105

  4. Weld-brazing of titanium. [resistance spot welding combined with brazing

    NASA Technical Reports Server (NTRS)

    Bales, T. T.; Royster, D. M.; Arnold, W. E., Jr.

    1973-01-01

    A joining process, designated weld-brazing, which combines resistance spot-welding and brazing has been developed at the NASA Langley Research Center. Resistance spot-welding is employed to position and aline the parts and to establish a suitable faying surface gap for brazing and contributes to the integrity of the joint. Brazing enhances the properties of the joint and reduces the stress concentrations normally associated with spotwelds. Ti-6Al-4V titanium alloy joints have been fabricated using 3003 aluminum braze both in a vaccum furnace and in a retort containing an inert gas environment.

  5. Brazed Joints Design and Allowables: Discuss Margins of Safety in Critical Brazed Structures

    NASA Technical Reports Server (NTRS)

    FLom, Yury

    2009-01-01

    This slide presentation tutorial discusses margins of safety in critical brazed structures. It reviews: (1) the present situation (2) definition of strength (3) margins of safety (4) design allowables (5) mechanical testing (6) failure criteria (7) design flowchart (8) braze gap (9) residual stresses and (10) delayed failures. This presentation addresses the strength of the brazed joints, the methods of mechanical testing, and our ability to evaluate the margins of safety of the brazed joints as it applies to the design of critical and expensive brazed assemblies.

  6. Rabs set the stage for polarity.

    PubMed

    Parker, Sara S; Cox, Christopher; Wilson, Jean M

    2017-01-26

    Cell polarity refers to the asymmetric localization of cellular components that allows cells to carry out their specialized functions, be they epithelial barrier function, transmission of action potentials in nerve cells, or modulation of the immune response. The establishment and maintenance of cell polarity requires the directed trafficking of membrane proteins and lipids - essential processes that are mediated by Rab GTPases. Interestingly, several of the Rabs that impact polarity are present in the earliest eukaryotes, and the Rab polarity repertoire has expanded as cells have become more complex. There is a substantial conservation of Rab function across diverse cell types. Rabs act through an assortment of effector proteins that include scaffolding proteins, cytoskeletal motors, and other small GTPases. In this review we highlight the similarities and differences in Rab function for the instruction of polarity in diverse cell types.

  7. Laser beam brazing of car body and aircraft components

    SciTech Connect

    Haferkamp, H.; Kreutzburg, K.

    1994-12-31

    At present, when brazing car body components for the automotive industry, manual flame brazing is mostly used. The advantage of brazing as compared to welding, is the lower hardness of the braze metal, making postmachining easier. But manual flame brazing also shows several main disadvantages, such as pores within the seam and a high thermal influence on the workpiece. Therefore, investigations on laser beam brazing concerning the reduction of the technological and economical disadvantages of the flame brazing process were carried out. Laser beam brazing of aluminum alloys is also a main topic of this presentation. The fundamental research in brazing mild steel was done on lap joints. The investigations about brazing mild steel and aluminum alloys have demonstrated that it is possible to braze these metals using laser beam radiation. Laser beam brazing of 3-dimensional mild steel components requires a special program for the brazing sequence, and new specifications in design and fabrication. But comparing seams made by laser beam brazing to manual flame brazing show that there are advantages to using the automated laser process. Laser beam brazing of aluminum alloys makes it possible to join metals with poor brazeability, although brazing conditions lead to a slight melting of the gap sides.

  8. Vacuum brazing beryllium to Monel

    SciTech Connect

    Glenn, T.G.; Grotsky, V.K.; Keller, D.L.

    1982-10-01

    The tensile strength of beryllium to Monel vacuum furnace brazed joints was studied. The filler used was the 72% Ag-28%Cu(BAg-8) alloy. The strength of these joints, which require the use of a titanium hydride powder or physical vapor deposited titanium wetting agent on the beryllium, was found to approach the yield strength of the base metals. Strength was found to be reduced by the interaction of increased titanium hydride quantity and brazing time. Metallographic and scanning electron microscope (SEM) studies correlated these effects with microstructure. The formation of the brittle copper-beryllium delta phase was found to require conditions of high brazing temperature and the presence of a reservoir of the copper-containing filler such as found in fillet areas. Two other filler metals: pure silver, and a 60% Ag-30% Cu-10%Sn (BAg-17) alloy were shown to be acceptable alternatives to the BAg-8 alloy in cases where the filler metal can be preplaced between the base metal surfaces.

  9. RAB13 — EDRN Public Portal

    Cancer.gov

    RAB13 is a member of the Rab family of small G proteins and plays a role in regulating membrane trafficking between trans-Golgi network (TGN) and recycling endosomes (RE). RAB13 is thought to participate in polarized transport, in the assembly and/or the activity of tight junctions, which are components of the apical junctional complex (AJC) of epithelial cells. There are several alternatively spliced transcript variants for this gene, and a pseudogene located on chromosome 12.

  10. Braze alloys for high temperature service

    NASA Technical Reports Server (NTRS)

    Lindberg, R. A.; Mckisson, R. L.; Erwin, G., Jr.

    1973-01-01

    Two groups of refractory metal compositions have been developed that are very useful as high temperature brazing alloys for sealing between ceramic and metal parts. Each group consists of various compositions of three selected refractory metals which, when combined, have characteristics required of good braze alloys.

  11. High-temperature nickel-brazing alloy

    NASA Technical Reports Server (NTRS)

    Powell, A. H.; Thompson, S. R.

    1970-01-01

    Gold-nickel brazing alloy, with 5 percent indium added to depress the melting point, is used for brazing of nickel-clad silver electrical conductors which operate at temperatures to 1200 deg F. Alloy has low resistivity, requires no flux, and is less corrosive than other gold-nickel, gold-copper alloys.

  12. Braze/Rebraze process for CRES steel

    NASA Technical Reports Server (NTRS)

    Silverman, C. E.

    1976-01-01

    Using induction brazing process with 8.5-Au/16.5-Cu/2.0-Ni braze alloy, joints in 21-6-9 CRES steel tubing can be reworked up to seven times, thus significantly reducing cost of fabrication, repair, and part replacement.

  13. Method for brazing together planar and nonplanar metal members

    DOEpatents

    Hammersand, Fred G.; Witkowski, Anthony J.

    1985-01-01

    The invention relates to a method and apparatus for brazing two metal members together, at least one of which is nonplanar, in a brazing furnace using a substantially pure brazing material. The method comprises the steps of utilizing a brazing fixture to hold the two metal members in tangential relation to one another along a portion of each member so that a cavity is formed adjacent to the contacting portions. A braze material is then positioned within the cavity. The braze fixture, the metal members, and the braze material are then placed in a brazing furnace. A heat shield is then placed over the braze fixture, the metal members, and the braze material to shield the braze material from direct furnace radiation. The furnace temperature is linearly increased at a rate of about 180.degree. C. per hour until a temperature of 350.degree. C. is achieved. Heat is transferred by conduction from the metal members to the braze material to cause the braze material to melt. Some material from the metal members slowly diffuses into the braze material forming a braze joint. The furnace is rapidly cooled to room temperature using nitrogen gas. The brazed assemblies made according to this method are superior to assemblies formed by heliarc welding.

  14. 46 CFR 56.30-30 - Brazed joints.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 2 2012-10-01 2012-10-01 false Brazed joints. 56.30-30 Section 56.30-30 Shipping COAST... Selection and Limitations of Piping Joints § 56.30-30 Brazed joints. (a) General (refer also to subpart 56.75). Brazed socket-type joints shall be made with suitable brazing alloys. The minimum socket...

  15. 46 CFR 56.30-30 - Brazed joints.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 2 2013-10-01 2013-10-01 false Brazed joints. 56.30-30 Section 56.30-30 Shipping COAST... Selection and Limitations of Piping Joints § 56.30-30 Brazed joints. (a) General (refer also to subpart 56.75). Brazed socket-type joints shall be made with suitable brazing alloys. The minimum socket...

  16. 46 CFR 56.30-30 - Brazed joints.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 2 2014-10-01 2014-10-01 false Brazed joints. 56.30-30 Section 56.30-30 Shipping COAST... Selection and Limitations of Piping Joints § 56.30-30 Brazed joints. (a) General (refer also to subpart 56.75). Brazed socket-type joints shall be made with suitable brazing alloys. The minimum socket...

  17. 46 CFR 56.30-30 - Brazed joints.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 2 2010-10-01 2010-10-01 false Brazed joints. 56.30-30 Section 56.30-30 Shipping COAST... Selection and Limitations of Piping Joints § 56.30-30 Brazed joints. (a) General (refer also to subpart 56.75). Brazed socket-type joints shall be made with suitable brazing alloys. The minimum socket...

  18. 46 CFR 56.30-30 - Brazed joints.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 2 2011-10-01 2011-10-01 false Brazed joints. 56.30-30 Section 56.30-30 Shipping COAST... Selection and Limitations of Piping Joints § 56.30-30 Brazed joints. (a) General (refer also to subpart 56.75). Brazed socket-type joints shall be made with suitable brazing alloys. The minimum socket...

  19. Are There Rab GTPases in Archaea?

    PubMed Central

    Surkont, Jaroslaw; Pereira-Leal, Jose B.

    2016-01-01

    A complex endomembrane system is one of the hallmarks of Eukaryotes. Vesicle trafficking between compartments is controlled by a diverse protein repertoire, including Rab GTPases. These small GTP-binding proteins contribute identity and specificity to the system, and by working as molecular switches, trigger multiple events in vesicle budding, transport, and fusion. A diverse collection of Rab GTPases already existed in the ancestral Eukaryote, yet, it is unclear how such elaborate repertoire emerged. A novel archaeal phylum, the Lokiarchaeota, revealed that several eukaryotic-like protein systems, including small GTPases, are present in Archaea. Here, we test the hypothesis that the Rab family of small GTPases predates the origin of Eukaryotes. Our bioinformatic pipeline detected multiple putative Rab-like proteins in several archaeal species. Our analyses revealed the presence and strict conservation of sequence features that distinguish eukaryotic Rabs from other small GTPases (Rab family motifs), mapping to the same regions in the structure as in eukaryotic Rabs. These mediate Rab-specific interactions with regulators of the REP/GDI (Rab Escort Protein/GDP dissociation Inhibitor) family. Sensitive structure-based methods further revealed the existence of REP/GDI-like genes in Archaea, involved in isoprenyl metabolism. Our analysis supports a scenario where Rabs differentiated into an independent family in Archaea, interacting with proteins involved in membrane biogenesis. These results further support the archaeal nature of the eukaryotic ancestor and provide a new insight into the intermediate stages and the evolutionary path toward the complex membrane-associated signaling circuits that characterize the Ras superfamily of small GTPases, and specifically Rab proteins. PMID:27034425

  20. Small GTPases Rab8a and Rab11a Are Dispensable for Rhodopsin Transport in Mouse Photoreceptors

    PubMed Central

    Ying, Guoxin; Gerstner, Cecilia D.; Frederick, Jeanne M.; Boye, Sanford L.; Hauswirth, William W.; Baehr, Wolfgang

    2016-01-01

    Rab11a and Rab8a are ubiquitous small GTPases shown as required for rhodopsin transport in Xenopus laevis and zebrafish photoreceptors by dominant negative (dn) disruption of function. Here, we generated retina-specific Rab11a (retRab11a) and Rab8a (retRab8a) single and double knockout mice to explore the consequences in mouse photoreceptors. Rhodopsin and other outer segment (OS) membrane proteins targeted correctly to OS and electroretinogram (ERG) responses in all three mutant mouse lines were indistinguishable from wild-type (WT). Further, AAV (adeno-associated virus)-mediated expression of dnRab11b in retRab11a-/- retina, or expression of dnRab8b in retRab8a-/- retina did not cause OS protein mislocalization. Finally, a retRab8a-/- retina injected at one month of age with AAVs expressing dnRab11a, dnRab11b, dnRab8b, and dnRab10 (four dn viruses on Rab8a-/- background) and harvested three months later exhibited normal OS protein localization. In contrast to results obtained with dnRab GTPases in Xenopus and zebrafish, mouse Rab11a and Rab8a are dispensable for proper rhodopsin and outer segment membrane protein targeting. Absence of phenotype after expression of four dn Rab GTPases in a Rab8a-/- retina suggests that Rab8b and Rab11b paralogs maybe dispensable as well. Our data thus demonstrate significant interspecies variation in photoreceptor membrane protein and rhodopsin trafficking. PMID:27529348

  1. RAB-6.1 and RAB-6.2 Promote Retrograde Transport in C. elegans.

    PubMed

    Zhang, Donglei; Dubey, Jyoti; Koushika, Sandhya P; Rongo, Christopher

    2016-01-01

    Retrograde transport is a critical mechanism for recycling certain membrane cargo. Following endocytosis from the plasma membrane, retrograde cargo is moved from early endosomes to Golgi followed by transport (recycling) back to the plasma membrane. The complete molecular and cellular mechanisms of retrograde transport remain unclear. The small GTPase RAB-6.2 mediates the retrograde recycling of the AMPA-type glutamate receptor (AMPAR) subunit GLR-1 in C. elegans neurons. Here we show that RAB-6.2 and a close paralog, RAB-6.1, together regulate retrograde transport in both neurons and non-neuronal tissue. Mutants for rab-6.1 or rab-6.2 fail to recycle GLR-1 receptors, resulting in GLR-1 turnover and behavioral defects indicative of diminished GLR-1 function. Loss of both rab-6.1 and rab-6.2 results in an additive effect on GLR-1 retrograde recycling, indicating that these two C. elegans Rab6 isoforms have overlapping functions. MIG-14 (Wntless) protein, which undergoes retrograde recycling, undergoes a similar degradation in intestinal epithelia in both rab-6.1 and rab-6.2 mutants, suggesting a broader role for these proteins in retrograde transport. Surprisingly, MIG-14 is localized to separate, spatially segregated endosomal compartments in rab-6.1 mutants compared to rab-6.2 mutants. Our results indicate that RAB-6.1 and RAB-6.2 have partially redundant functions in overall retrograde transport, but also have their own unique cellular- and subcellular functions.

  2. RAB-6.1 and RAB-6.2 Promote Retrograde Transport in C. elegans

    PubMed Central

    Zhang, Donglei; Dubey, Jyoti; Koushika, Sandhya P.; Rongo, Christopher

    2016-01-01

    Retrograde transport is a critical mechanism for recycling certain membrane cargo. Following endocytosis from the plasma membrane, retrograde cargo is moved from early endosomes to Golgi followed by transport (recycling) back to the plasma membrane. The complete molecular and cellular mechanisms of retrograde transport remain unclear. The small GTPase RAB-6.2 mediates the retrograde recycling of the AMPA-type glutamate receptor (AMPAR) subunit GLR-1 in C. elegans neurons. Here we show that RAB-6.2 and a close paralog, RAB-6.1, together regulate retrograde transport in both neurons and non-neuronal tissue. Mutants for rab-6.1 or rab-6.2 fail to recycle GLR-1 receptors, resulting in GLR-1 turnover and behavioral defects indicative of diminished GLR-1 function. Loss of both rab-6.1 and rab-6.2 results in an additive effect on GLR-1 retrograde recycling, indicating that these two C. elegans Rab6 isoforms have overlapping functions. MIG-14 (Wntless) protein, which undergoes retrograde recycling, undergoes a similar degradation in intestinal epithelia in both rab-6.1 and rab-6.2 mutants, suggesting a broader role for these proteins in retrograde transport. Surprisingly, MIG-14 is localized to separate, spatially segregated endosomal compartments in rab-6.1 mutants compared to rab-6.2 mutants. Our results indicate that RAB-6.1 and RAB-6.2 have partially redundant functions in overall retrograde transport, but also have their own unique cellular- and subcellular functions. PMID:26891225

  3. Braze alloy spreading on steel

    NASA Technical Reports Server (NTRS)

    Siewert, T. A.; Heine, R. W.; Lagally, M. G.

    1978-01-01

    Scanning electron microscopy (SEM) and Auger electron microscopy (AEM) were employed to observe elemental surface decomposition resulting from the brazing of a copper-treated steel. Two types of steel were used for the study, stainless steel (treated with a eutectic silver-copper alloy), and low-carbon steel (treated with pure copper). Attention is given to oxygen partial pressure during the processes; a low enough pressure (8 x 10 to the -5th torr) was found to totally inhibit the spreading of the filler material at a fixed heating cycle. With both types of steel, copper treatment enhanced even spreading at a decreased temperature.

  4. RAB21 Activity Assay Using GST-fused APPL1

    PubMed Central

    Jean, Steve; Kiger, Amy A.

    2016-01-01

    The Rab family of small GTPases are essential regulators of membrane trafficking events. As with other small GTPase families, Rab GTPases cycle between an inactive GDP-bound state and an active GTP-bound state. Guanine nucleotide exchange factors (GEFs) promote Rab activation with the exchange of bound GDP for GTP, while GTPase-activating proteins (GAPs) regulate Rab inactivation with GTP hydrolysis. Numerous methods have been established to monitor the activation status of Rab GTPases. Of those, FRET-based methods are used to identify when and where a Rab GTPase is activated in cells. Unfortunately, the generation of such probes is complex, and only a limited number of Rabs have been probed this way. Biochemical purification of activated Rabs from cell or tissue extracts is easily achievable through the use of a known Rab effector domain to pull down a specific GTP-bound Rab form. Although this method is not ideal for detailed subcellular localization, it can offer temporal resolution of Rab activity. The identification of a growing number of specific effectors now allows tests for activation levels of many Rab GTPases in specific conditions. Here, we described an affinity purification approach using GST fused APPL1 (a known RAB21 effector) to test RAB21 activation in mammalian cells. This method was successfully used to assay changes in RAB21 activation status under nutrient rich versus starved conditions and to test the requirement of the MTMR13 RAB21 GEF in this process. PMID:28251173

  5. Theory and modeling of active brazing.

    SciTech Connect

    van Swol, Frank B.; Miller, James Edward; Lechman, Jeremy B.; Givler, Richard C.

    2013-09-01

    Active brazes have been used for many years to produce bonds between metal and ceramic objects. By including a relatively small of a reactive additive to the braze one seeks to improve the wetting and spreading behavior of the braze. The additive modifies the substrate, either by a chemical surface reaction or possibly by alloying. By its nature, the joining process with active brazes is a complex nonequilibrium non-steady state process that couples chemical reaction, reactant and product diffusion to the rheology and wetting behavior of the braze. Most of the these subprocesses are taking place in the interfacial region, most are difficult to access by experiment. To improve the control over the brazing process, one requires a better understanding of the melting of the active braze, rate of the chemical reaction, reactant and product diffusion rates, nonequilibrium composition-dependent surface tension as well as the viscosity. This report identifies ways in which modeling and theory can assist in improving our understanding.

  6. PpRab1, a Rab GTPase from maritime pine is differentially expressed during embryogenesis.

    PubMed

    Gonçalves, Sónia; Cairney, John; Rodríguez, María Pérez; Cánovas, Francisco; Oliveira, Margarida; Miguel, Célia

    2007-09-01

    Rab-related small GTP-binding proteins are known to be involved in the regulation of the vesicular transport system in eukaryotic cells. We report the characterization of a previously isolated full-length cDNA PpRab1 from Pinus pinaster. Amino acid sequence analysis revealed the presence of G1-G5 conserved domains of the GTPase Ras superfamily and a double cysteine motif in the C-terminal, characteristic of Rab proteins. The PpRab1 protein shows high sequence similarity to several Rab1 GTP-binding proteins in plants. Phylogenetic analysis showed that, within the Ras superfamily, PpRab1 is more closely related to the Rab family and within this, PpRab1 protein was found to cluster with Arabidopsis subfamily AtRABE, whose members are known to regulate ER-to-Golgi membrane trafficking steps. PpRab1 transcripts were expressed at constitutively high levels for the initial stages of zygotic embryo development, and then their relative abundance decreased as embryo matures. The PpRab1 transcript is not embryo-specific as it was found in roots, cotyledons and hypocotyls. An increase in PpRab1 expression level was observed when seeds are germinated and collected at successive time points of development. In situ RT-PCR analysis revealed an expression signal in early zygotic embryos. In view of the proposed roles of Rab1 GTP-binding protein, the possible function of the protein encoded by PpRab1 in embryogenesis is discussed.

  7. The Rab interacting lysosomal protein (RILP) homology domain functions as a novel effector domain for small GTPase Rab36: Rab36 regulates retrograde melanosome transport in melanocytes.

    PubMed

    Matsui, Takahide; Ohbayashi, Norihiko; Fukuda, Mitsunori

    2012-08-17

    Small GTPase Rab functions as a molecular switch that drives membrane trafficking through specific interaction with its effector molecule. Thus, identification of its specific effector domain is crucial to revealing the molecular mechanism that underlies Rab-mediated membrane trafficking. Because of the large numbers of Rab isoforms in higher eukaryotes, however, the effector domains of most of the vertebrate- or mammalian-specific Rabs have yet to be determined. In this study we screened for effector molecules of Rab36, a previously uncharacterized Rab isoform that is largely conserved in vertebrates, and we succeeded in identifying nine Rab36-binding proteins, including RILP (Rab interacting lysosomal protein) family members. Sequence comparison revealed that five of nine Rab36-binding proteins, i.e. RILP, RILP-L1, RILP-L2, and JIP3/4, contain a conserved coiled-coil domain. We identified the coiled-coil domain as a RILP homology domain (RHD) and characterized it as a common Rab36-binding site. Site-directed mutagenesis of the RHD of RILP revealed the different contributions by amino acids in the RHD to binding activity toward Rab7 and Rab36. Expression of RILP in melanocytes, but not expression of its Rab36 binding-deficient mutants, induced perinuclear aggregation of melanosomes, and this effect was clearly attenuated by knockdown of endogenous Rab36 protein. Moreover, knockdown of Rab36 in Rab27A-deficient melanocytes, which normally exhibit perinuclear melanosome aggregation because of increased retrograde melanosome transport activity, caused dispersion of melanosomes from the perinucleus to the cell periphery, but knockdown of Rab7 did not. Our findings indicated that Rab36 mediates retrograde melanosome transport in melanocytes through interaction with RILP.

  8. Tyrosine phosphorylation of Rab7 by Src kinase.

    PubMed

    Lin, Xiaosi; Zhang, Jiaming; Chen, Lingqiu; Chen, Yongjun; Xu, Xiaohui; Hong, Wanjin; Wang, Tuanlao

    2017-03-20

    The small molecular weight GTPase Rab7 is a key regulator for late endosomal/lysosomal membrane trafficking, it was known that Rab7 is phosphorylated, but the corresponding kinase and the functional regulation of Rab7 phosphorylation remain unclear. We provide evidence here that Rab7 is a substrate of Src kinase, and is tyrosine-phosphorylated by Src, withY183 residue of Rab7 being the optimal phosphorylation site for Src. Further investigations demonstrated that the tyrosine phosphorylation of Rab7 depends on the guanine nucleotide binding activity of Rab7 and the activity of Src kinase. The tyrosine phosphorylation of Rab7 is physiologically induced by EGF, and impairs the interaction of Rab7 with RILP, consequently inhibiting EGFR degradation and sustaining Akt signaling. These results suggest that the tyrosine phosphorylation of Rab7 may be involved in coordinating membrane trafficking and cell signaling.

  9. Development of rapidly quenched brazing foils to join tungsten alloys with ferritic steel

    NASA Astrophysics Data System (ADS)

    Kalin, B. A.; Fedotov, V. T.; Sevrjukov, O. N.; Moeslang, A.; Rohde, M.

    2004-08-01

    Results on rapidly solidified filler metals for tungsten brazing are presented. A rapidly quenched foil-type filler metal based on Ni bal-15Cr-4Mo-4Fe-(0.5-1.0)V-7.5Si-1.5B was developed to braze tungsten to ferritic/martensitic Crl3Mo2NbVB steel (FS) for helium gas cooled divertors and plasma facing components. Polycrystalline W-2CeO 2 and monocrystalline pure tungsten were brazed to the steel under vacuum at 1150 °C, using a 0.5 mm thick foil spacer made of a 50Fe-50Ni alloy. As a result of thermocycling tests (100 cycles between 700 °C/20 min and air-water cooling/3-5 min) on brazed joints, tungsten powder metallurgically processed W-2CeO 2 failed due to residual stresses, whereas the brazed joint with zone-melted monocrystalline tungsten withstood the thermocycling tests.

  10. Brazing of Stainless Steels to Yttria Stabilized Zirconia (YSZ) for Solid Oxide Fuel Cells

    NASA Technical Reports Server (NTRS)

    Shpargel, Tarah P.; Needham, Robert J.; Singh, M.; Kung, Steven C.

    2005-01-01

    Recently, there has been a great deal of interest in research, development, and commercialization of solid oxide fuel cells. Joining and sealing are critical issues that will need to be addressed before SOFC's can truly perform as expected. Ceramics and metals can be difficult to join together, especially when the joint must withstand up to 900 C operating temperature of the SOFC's. The goal of the present study is to find the most suitable braze material for joining of yttria stabilized zirconia (YSZ) to stainless steels. A number of commercially available braze materials TiCuSil, TiCuNi, Copper-ABA, Gold-ABA, and Gold-ABA-V have been evaluated. The oxidation behavior of the braze materials and steel substrates in air was also examined through thermogravimetric analysis. The microstructure and composition of the brazed regions have been examined by optical and scanning electron microscopy and EDS analysis. Effect of braze composition and processing conditions on the interfacial microstructure and composition of the joint regions will be presented.

  11. Failure Assessment Diagram for Titanium Brazed Joints

    NASA Technical Reports Server (NTRS)

    Flom, Yury; Jones, Justin S.; Powell, Mollie M.; Puckett, David F.

    2011-01-01

    The interaction equation was used to predict failure in Ti-4V-6Al joints brazed with Al 1100 filler metal. The joints used in this study were geometrically similar to the joints in the brazed beryllium metering structure considered for the ATLAS telescope. This study confirmed that the interaction equation R(sub sigma) + R(sub Tau) = 1, where R(sub sigma) and R(sub Tau)are normal and shear stress ratios, can be used as conservative lower bound estimate of the failure criterion in ATLAS brazed joints as well as for construction of the Failure Assessment Diagram (FAD).

  12. Direct metal brazing to cermet feedthroughs

    DOEpatents

    Not Available

    1982-07-29

    An improved method for brazing metallic components to a cermet surface in an alumina substrate eliminates the prior art metallized layer over the cermet via and adjoining alumina surfaces. Instead, a nickel layer is applied over the cermet surface only and metallic components are brazed directly to this nickel coated cermet surface. As a result, heretofore unachievable tensile strength joints are produced. In addition, cermet vias with their brazed metal components can be spaced more closely in the alumina substrate because of the elimination of the prior art metallized alumina surfaces.

  13. Direct metal brazing to cermet feedthroughs

    DOEpatents

    Hopper, Jr., Albert C.

    1984-12-18

    An improved method for brazing metallic components to a cermet surface in an alumina substrate eliminates the prior art metallized layer over the cermet via and adjoining alumina surfaces. Instead, a nickel layer is applied over the cermet surface only and metallic components are brazed directly to this nickel coated cermet surface. As a result, heretofore unachievable tensile strength joints are produced. In addition, cermet vias with their brazed metal components can be spaced more closely in the alumina substrate because of the elimination of the prior art metallized alumina surfaces.

  14. Infrared repair brazing of 403 stainless steel with a nickel-based braze alloy

    NASA Astrophysics Data System (ADS)

    Shiue, R. K.; Wu, S. K.; Hung, C. M.

    2002-06-01

    Martensitic stainless steel (403SS) is extensively used for intermediate and low-pressure steam turbine blades in fossil-fuel power plants. The purpose of this investigation is to study the repair of shallow cracks on the surface of 403SS steam turbine blades by infrared repair brazing using rapid thermal cycles. A nickel-based braze alloy (NICROBRAZ LM) is used as filler metal. The braze alloy after brazing is primarily comprised of borides and an FeNi3 matrix with different amounts of alloying elements, especially B and Si. As the brazing temperature increases, more Fe atoms are dissolved into the molten braze. Some boron atoms diffuse into the 403SS substrate primarily via grain boundary diffusion and form B-Cr-Fe intermetallic precipitates along the grain boundaries. The LM filler metal demonstrates better performance than 403SS in both microhardness and wear tests. It is also noted that specimens brazed in a vacuum have less porosity than those brazed in an Ar atmosphere. The shear strength of the joint is around 300 MPa except for specimens brazed in short time periods, e.g., 5 seconds in Ar flow and 30 seconds in vacuum. The fractographs mainly consist of brittle fractures and no ductile dimple fractures observed in the scanning electron microscope (SEM) examination.

  15. The RCP-Rab11 complex regulates endocytic protein sorting.

    PubMed

    Peden, Andrew A; Schonteich, Eric; Chun, John; Junutula, Jagath R; Scheller, Richard H; Prekeris, Rytis

    2004-08-01

    Rab 11 GTPase is an important regulator of endocytic membrane traffic. Recently, we and others have identified a novel family of Rab11 binding proteins, known as Rab11-family interacting proteins (FIPs). One of the family members, Rab coupling protein (RCP), was identified as a protein binding to both Rab4 and Rab11 GTPases. RCP was therefore suggested to serve a dual function as Rab4 and Rab11 binding protein. In this study, we characterized the cellular functions of RCP and mapped its interactions with Rab4 and Rab11. Our data show that RCP interacts only weakly with Rab4 in vitro and does not play the role of coupling Rab11 and Rab4 in vivo. Furthermore, our data indicate that the RCP-Rab11 complex regulates the sorting of transferrin receptors from the degradative to the recycling pathway. We therefore propose that RCP functions primarily as a Rab11 binding protein that regulates protein sorting in tubular endosomes.

  16. RAB5A — EDRN Public Portal

    Cancer.gov

    From UniProtKB/Swiss-Prot: The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different sets of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion. RAB5A is required for the fusion of plasma membranes and early endosomes. Contributes to the regulation of filopodia extension.

  17. Thermal resilient multiple jaw braze fixture

    DOEpatents

    Ney, R.; Perrone, A.J.

    1995-07-11

    A braze fixture has side walls forming a cavity with an opening to receive a stack of parts to be brazed. Sidewalls of the housing have a plurality of bearing receiving openings into which bearing rods or jaws are inserted to align the stacked elements of the workpiece. The housing can also have view ports to allow a visual check of the alignment. Straps or wires around the fixture are selected to have thermal characteristics similar to the thermal characteristics of the workpiece undergoing brazing. The straps or wires make physical contact with the bearing rods thereby causing bearing rods to maintain the workpiece in proper alignment throughout the entire brazing cycle. 9 figs.

  18. The Ccz1-Mon1-Rab7 module and Rab5 control distinct steps of autophagy

    PubMed Central

    Hegedűs, Krisztina; Takáts, Szabolcs; Boda, Attila; Jipa, András; Nagy, Péter; Varga, Kata; Kovács, Attila L.; Juhász, Gábor

    2016-01-01

    The small GTPase Rab5 promotes recruitment of the Ccz1-Mon1 guanosine exchange complex to endosomes to activate Rab7, which facilitates endosome maturation and fusion with lysosomes. How these factors function during autophagy is incompletely understood. Here we show that autophagosomes accumulate due to impaired fusion with lysosomes upon loss of the Ccz1-Mon1-Rab7 module in starved Drosophila fat cells. In contrast, autophagosomes generated in Rab5-null mutant cells normally fuse with lysosomes during the starvation response. Consistent with that, Rab5 is dispensable for the Ccz1-Mon1–dependent recruitment of Rab7 to PI3P-positive autophagosomes, which are generated by the action of the Atg14-containing Vps34 PI3 kinase complex. Finally, we find that Rab5 is required for proper lysosomal function. Thus the Ccz1-Mon1-Rab7 module is required for autophagosome-lysosome fusion, whereas Rab5 loss interferes with a later step of autophagy: the breakdown of autophagic cargo within lysosomes. PMID:27559127

  19. Joining of alumina by vacuum brazing

    NASA Astrophysics Data System (ADS)

    Heikinheimo, Liisa; Siren, Mika; Kauppinen, Pentti

    1993-08-01

    The active brazing method for diffusion bonding of ceramics to metals is addressed. This method is very flexible compared to the traditional Mo-manganese coating with subsequent brazing that includes four process steps: in active brazing the process is done in one step. The joint properties are favorable, the residual stress build up is limited if the braze is correctly selected and the thermal cycle is controlled, and the resulting strength and leak tightness are good. In experimental work the joinability of alumina to titanium and Ni superalloys was studied by wetting experiments, nondestructive test and shear strength measurements. The spreading of the braze is affected not only by the surface conditions of mating materials but also by the type of the brazing alloy. The Ag-Cu base alloys give better wetting, strength and leak tightness properties than the Ag base alloys. A shear test method was developed for the mechanical testing of metal-ceramic joints. However, the sample geometry affects the measured values, namely a smaller specimen size provides better results. The correlation between the C-SAM results, which describe the ratio between the true bonded area and unbonded area, and measured shear strength values is presented. The dependence between the measured strength and the area of the joint defects becomes obvious and should be studied in more detail.

  20. Ceramic-Metal Brazing, from Fundamentals to Applications: A Review of Sandia National Laboratories Brazing Capabilities, Needs, and Opportunities

    NASA Astrophysics Data System (ADS)

    Hosking, F. M.; Stephens, J. J.; Glass, S. J.; Johannes, J. E.; Kotula, P.

    2002-05-01

    The purpose of the report is to summarize discussions from a Ceramic/Metal Brazing: From Fundamentals to Applications Workshop that was held at Sandia National Laboratories in Albuquerque, NM on April 4, 2001. Brazing experts and users who bridge common areas of research, design, and manufacturing participated in the exercise. External perspectives on the general state of the science and technology for ceramics and metal brazing were given. Other discussions highlighted and critiqued Sandia's brazing research and engineering programs, including the latest advances in braze modeling and materials characterization. The workshop concluded with a facilitated dialogue that identified critical brazing research needs and opportunities.

  1. A complete Rab screening reveals novel insights in Weibel-Palade body exocytosis.

    PubMed

    Zografou, Sofia; Basagiannis, Dimitris; Papafotika, Alexandra; Shirakawa, Ryutaro; Horiuchi, Hisanori; Auerbach, Daniel; Fukuda, Mitsunori; Christoforidis, Savvas

    2012-10-15

    Weibel-Palade bodies (WPBs) are endothelial-cell-specific organelles that, upon fusion with the plasma membrane, release cargo molecules that are essential in blood vessel abnormalities, such as thrombosis and inflammation, as well as in angiogenesis. Despite the importance of WPBs, the basic mechanisms that mediate their secretion are only poorly understood. Rab GTPases play fundamental role in the trafficking of intracellular organelles. Yet, the only known WPB-associated Rabs are Rab27a and Rab3d. To determine the full spectrum of WPB-associated Rabs we performed a complete Rab screening by analysing the localisation of all Rabs in WPBs and their involvement in the secretory process in endothelial cells. Apart from Rab3 and Rab27, we identified three additional Rabs, Rab15 (a previously reported endocytic Rab), Rab33 and Rab37, on the WPB limiting membrane. A knockdown approach using siRNAs showed that among these five WPB Rabs only Rab3, Rab27 and Rab15 are required for exocytosis. Intriguingly, we found that Rab15 cooperates with Rab27a in WPB secretion. Furthermore, a specific effector of Rab27, Munc13-4, appears to be also an effector of Rab15 and is required for WPB exocytosis. These data indicate that WPB secretion requires the coordinated function of a specific group of Rabs and that, among them, Rab27a and Rab15, as well as their effector Munc13-4, cooperate to drive exocytosis.

  2. Brazing of Stainless Steels to Yttria Stabilized Zirconia (YSZ) Using Silver -Base Brazes

    NASA Technical Reports Server (NTRS)

    Singh, Mrityunjay; Shpargel, Tarah P.; Asthana, Rajiv

    2005-01-01

    Three silver-base brazes containing either noble metal palladium (Palcusil-10 and Palcusil-15) or active metal titanium (Ticusil) were evaluated for high-temperature oxidation resistance, and their effectiveness in joining yttria stabilized zirconia (YSZ) to a corrosion-resistant ferritic stainless steel. Thermogravimetric analysis (TGA), and optical- and scanning electron microscopy (SEM) coupled with energy dispersive spectrometry (EDS) were used to evaluate the braze oxidation behavior and the structure and chemistry of the YSZ/braze/steel joints. The effect of the braze type and processing conditions on the interfacial microstructure and composition of the joint regions is discussed with reference to the chemical changes that occur at the interface. It was found that chemical interdiffusion of the constituents of YSZ, steel and the brazes led to compositional changes and/or interface reconstruction, and metallurgically sound joints.

  3. Thousands of rab GTPases for the cell biologist.

    PubMed

    Diekmann, Yoan; Seixas, Elsa; Gouw, Marc; Tavares-Cadete, Filipe; Seabra, Miguel C; Pereira-Leal, José B

    2011-10-01

    Rab proteins are small GTPases that act as essential regulators of vesicular trafficking. 44 subfamilies are known in humans, performing specific sets of functions at distinct subcellular localisations and tissues. Rab function is conserved even amongst distant orthologs. Hence, the annotation of Rabs yields functional predictions about the cell biology of trafficking. So far, annotating Rabs has been a laborious manual task not feasible for current and future genomic output of deep sequencing technologies. We developed, validated and benchmarked the Rabifier, an automated bioinformatic pipeline for the identification and classification of Rabs, which achieves up to 90% classification accuracy. We cataloged roughly 8.000 Rabs from 247 genomes covering the entire eukaryotic tree. The full Rab database and a web tool implementing the pipeline are publicly available at www.RabDB.org. For the first time, we describe and analyse the evolution of Rabs in a dataset covering the whole eukaryotic phylogeny. We found a highly dynamic family undergoing frequent taxon-specific expansions and losses. We dated the origin of human subfamilies using phylogenetic profiling, which enlarged the Rab repertoire of the Last Eukaryotic Common Ancestor with Rab14, 32 and RabL4. Furthermore, a detailed analysis of the Choanoflagellate Monosiga brevicollis Rab family pinpointed the changes that accompanied the emergence of Metazoan multicellularity, mainly an important expansion and specialisation of the secretory pathway. Lastly, we experimentally establish tissue specificity in expression of mouse Rabs and show that neo-functionalisation best explains the emergence of new human Rab subfamilies. With the Rabifier and RabDB, we provide tools that easily allows non-bioinformaticians to integrate thousands of Rabs in their analyses. RabDB is designed to enable the cell biology community to keep pace with the increasing number of fully-sequenced genomes and change the scale at which we perform

  4. Design of a braze alloy for fast epitaxial brazing of superalloys

    NASA Astrophysics Data System (ADS)

    Piegert, S.; Laux, B.; Rösier, J.

    2012-07-01

    For the repair of directionally solidified turbine components made of nickel-based superalloys, a new high-temperature brazing method has been developed. Utilising heterogeneous nucleation on the crack surface, the microstructure of the base material can be reproduced, i.e. single crystallinity can be maintained. In contrast to commonly used eutectic braze alloys, such as nickel-boron or nickel-silicon systems, the process is not diffusion controlled but works with a consolute binary base system. The currently applied epitaxial brazing methods rely on isothermal solidification diffusing the melting point depressants into the base material until their concentration is reduced so that the liquid braze solidifies. Contrary, the identified Ni-Mn consolute system enables a temperature driven epitaxial solidification resulting in substantially reduced process duration. The development of the braze alloys was assisted using the CALPHAD software Thermo-Calc. The solidification behaviour was estimated by kinetic calculations with realistic boundary conditions. Finally, the complete system, including braze alloy as well as substrate material, was modelled by means of DICTRA. Subsequently, the thermodynamic properties of the braze alloys were experimentally analysed by DSC measurements. For brazing experiments 300 μm wide parallel gaps were used. Complete epitaxial solidification, i.e. the absence of high-angle grain boundaries, could be achieved within brazing times being up to two orders of magnitude shorter compared to diffusion brazing processes. Theoretically and experimentally evaluated process windows reveal similar shapes. However, a distinct shift has to be stated which can be ascribed to the limited accuracy of the underlying thermodynamic databases.

  5. Aluminum core structures brazed without use of flux

    NASA Technical Reports Server (NTRS)

    1966-01-01

    Aluminum alloy face sheets are brazed to aluminum alloy honeycomb cores without using corrosive flux by means of one or three methods. The completed brazed structure has the high-strength characteristics of heat treated aluminum alloys.

  6. Refractory metals welded or brazed with tungsten inert gas equipment

    NASA Technical Reports Server (NTRS)

    Wisner, J. P.

    1965-01-01

    Appropriate brazing metals and temperatures facilitate the welding or brazing of base metals with tungsten inert gas equipment. The highest quality bond is obtained when TIG welding is performed in an inert atmosphere.

  7. High-strength braze joints between copper and steel

    NASA Technical Reports Server (NTRS)

    Kuhn, R. F.

    1967-01-01

    High-strength braze joints between copper and steel are produced by plating the faying surface of the copper with a layer of gold. This reduces porosity in the braze area and strengthens the resultant joint.

  8. Ultrasonics permits brazing complex stainless steel assembly without flux

    NASA Technical Reports Server (NTRS)

    Baker, W. H.

    1967-01-01

    Ultrasonic vibration of an assembly of stainless steel instrumentation tubes ensures brazing without flux. Vibration with an ultrasonic transducer permits the brazing material to flow down each tube in contact with a seal plug installed in a pressure vessel wall.

  9. Isolation, sequence identification and expression profile of three novel genes Rab2A, Rab3A and Rab7A from Black-boned sheep (Ovis aries).

    PubMed

    He, Y D; Liu, D D; Xi, D M; Yang, L Y; Tan, Y W; Liu, Q; Mao, H M; Deng, W D

    2010-01-01

    Complete coding sequences of three Black-boned sheep (Ovis aries) genes Rab2A, Rab3A and Rab7A were amplified using reverse transcription polymerase chain reaction (RT-PCR) based on the conserved sequence information of cattle or other mammals known to be highly homologous to sheep ESTs. The Black-boned sheep Rab2A gene encodes a protein of 226 amino acids which contains the conserved putative RabL2 domain and is highly homologous to the Rab2A proteins of seven other species--cattle (96%), human (83%), Sumatran orangutan (82%), rat (81%), mouse (80%), African clawed frog (72%) and zebrafish (71%). The Black-boned sheep Rab3A gene encodes a protein of 220 amino acids that contains the conserved putative Rab3 domain and is very similar to the Rab3A proteins of four species--cattle (99%), African clawed frog (99%), Western clawed frog (98%) and zebrafish (95%). And the Black-boned sheep Rab7A gene encodes a protein of 207 amino acids that contains the conserved putative Rab7 domain and has high homology with the Rab7A proteins of six other species--human (99%), dog (99%), Sumatran orangutan (99%), zebrafish (97%), rabbit (97%) and African clawed frog (96%). Analysis of the phylogenetic tree has demonstrated that the Black-boned sheep Rab2A, Rab3A and Rab7A proteins share a common ancestor and the tissue expression analysis has shown that the corresponding genes are expressed in a range of tissues including leg muscle, kidney, skin, longissimus dorsi muscle, spleen, heart and liver. Our experiment is the first to provide the primary foundation for a further insight into these three sheep genes.

  10. Distinct yet overlapping roles of Rab GTPases on synaptic vesicles

    PubMed Central

    Pavlos, Nathan J

    2011-01-01

    Exo-endocytotic cycling of synaptic vesicles (SVs) is one of the most intensely studied membrane trafficking pathways. It is governed by sets of conserved proteins including Rab GTPases. Long considered to define the identity and composition of a subcellular organelle, it has become increasingly evident that multiple Rabs co-exist on intracellular compartments, each contributing to its membrane organization and specialised function. Indeed, we have recently demonstrated that at least 11 distinct Rab proteins co-exist on highly purified SVs. These include Rabs involved in exocytosis (Rab3a/b/c and Rab27b) and intermediates of SV recycling such as early endosomes (Rab4, Rab5, Rab10, Rab11b and Rab14). Interestingly, we found that while two of these proteins, namely Rab3a and Rab27b, exhibited differential cycling dynamics on SV membranes; they played complementary roles during Ca2+-triggered neurotransmitter release. The implications of these findings in the SV trafficking cycle are discussed. PMID:21776405

  11. Braze Process Optimization Involving Conventional Metal/Ceramic Brazing with 50Au-50Cu Alloy

    SciTech Connect

    MALIZIA JR.,LOUIS A.; MEREDITH,KEITH W.; APPEL,DANIEL B.; MONROE,SAUNDRA L.; BURCHETT,STEVEN N.; STEPHENS JR.,JOHN J.

    1999-12-15

    Numerous process variables can influence the robustness of conventional metal/ceramic brazing processes. Experience with brazing of hermetic vacuum components has identified the following parameters as influencing the outcome of hydrogen furnace brazed Kovar{trademark} to metallized alumina braze joints: (a) Mo-Mn metallization thickness, sinter fire temperature and porosity (b) Nil plate purity, thickness, and sinter firing conditions (c) peak process temperature, time above liquidus and (d) braze alloy washer thickness. ASTM F19 tensile buttons are being used to investigate the above parameters. The F19 geometry permits determination of both joint hermeticity and tensile strength. This presentation will focus on important lessons learned from the tensile button study: (A) the position of the Kovar{trademark} interlayer can influence the joint tensile strength achieved--namely, off-center interlayers can lead to residual stress development in the ceramic and degrade tensile strength values. Finite element analysis has been used to demonstrate the expected magnitude in strength degradation as a function of misalignment. (B) Time above liquidus (TAL) and peak temperature can influence the strength and alloying level of the resulting braze joint. Excessive TAL or peak temperatures can lead to overbraze conditions where all of the Ni plate is dissolved. (C) Metallize sinter fire processes can influence the morphology and strength obtained from the braze joints.

  12. Brazing of dispersion-strengthened aluminum

    SciTech Connect

    Bjoerneklett, B.; Grong, O.; Anisdahl, L.; Hellum, E.; Sande, V.

    1996-03-01

    In recent years, the use of rapid solidification powder metallurgy has made it possible to develop a new family of aluminum alloys exhibiting unique properties. One of these materials, dispersion-strengthened (DS) aluminum, is currently being produced for commercial purposes at Raufoss Technology AS, Norway. Dispersion-strengthened aluminum derives its high strength from nanoscale AlN particles embedded in an aluminum matrix. DS Al is expected to be well suited as construction material for high-temperature applications where weight reductions are of particular concern. The present investigation has focused on the wetting behavior of DS aluminum under conditions applicable to brazing. The results from the Sessile drop experiments show that a eutectic Al-Si brazing alloy will completely wet the base metal both under high-vacuum conditions and in controlled argon atmospheres, provided that the partial pressure of oxygen is sufficiently low. The main problem appears to be the stability of the matrix grain structure. In general, the process of grain erosion and coarsening can be controlled by restricting the supply of the brazing alloy so that only a small metal volume is exposed to erosion. In addition, there is a great potential for reducing the thermodynamic driving force of the erosion reaction by proper adjustments of the brazing alloy composition and/or the brazing temperature. Sill, grain boundary liquidation may be a problem which, in turn, may require additions of surface active elements to the filler metal to control the wetting behavior.

  13. Graphite-metal brazing for thermal applications

    SciTech Connect

    Hosking, F.M.; Koski, J.A.

    1991-01-01

    Various plasma facing components are being designed and fabricated to support Magnetic Fusion Energy experiments. They typically consist of graphite tiles mechanically or metallurgically attached to metallic cooling substrates. This paper will discuss the active brazing of isotropic and pyrolytic graphite to oxygen free, high conductivity (OFHC) Cu and an alumina-dispersion strengthened Cu with a Ag-Cu-Ti active filler metal. The Ti constituent promotes direct wetting of graphite with the formation of a thin TiC reaction layer. Joint design and materials selection are critical factors since graphite and Cu have large thermal expansion differences that affect residual stresses after brazing and subsequent component thermal performance. Low thermal expansion Mo and compliant Cu interlayers were introduced to lower the residual stresses and extend the thermal life of prototype graphite-Cu braze joints. Although the interlayers showed evidence of reducing the incidence of graphite cracking and spalling under thermal loading when brazed to the dispersion-stengthened Cu, the best graphite braze joints were produced with the more ductile OFHC Cu substrates and no interlayer. These latter joints survived simulated tokamak surface high heat fluxes of 30 MW{center dot}m{sup {minus}2} or greater, while comparable dispersion strengthened Cu samples failed at 10 MW{center dot}m{sup {minus}2}. 23 refs., 14 figs., 2 tabs.

  14. Graphite-metal brazing for thermal applications

    NASA Astrophysics Data System (ADS)

    Hosking, F. M.; Koski, J. A.

    Various plasma facing components are being designed and fabricated to support Magnetic Fusion Energy experiments. They typically consist of graphite tiles mechanically or metallurgically attached to metallic cooling substrates. This paper will discuss the active brazing of isotropic and pyrolytic graphite to oxygen free, high conductivity (OFHC) Cu and an alumina-dispersion strengthened Cu with a Ag-Cu-Ti active filler metal. The Ti constituent promotes direct wetting of graphite with the formation of a thin TiC reaction layer. Joint design and materials selection are critical factors since graphite and Cu have large thermal expansion differences that affect residual stresses after brazing and subsequent component thermal performance. Low thermal expansion Mo and compliant Cu interlayers were introduced to lower the residual stresses and extend the thermal life of prototype graphite-Cu braze joints. Although the interlayers showed evidence of reducing the incidence of graphite cracking and spalling under thermal loading when brazed to the dispersion stengthened Cu, the best graphite braze joints were produced with the more ductile OFHC Cu substrates and no interlayer. These latter joints survived simulated tokamak surface high heat fluxes of 30 MW x m(exp -2) or greater, while comparable dispersion strengthened Cu samples failed at 10 MW x m(exp -2)

  15. Structural Insights into Rab27 Recruitment by its Effectors

    NASA Astrophysics Data System (ADS)

    M. G. Chavas, Leonard; Ihara, Kentaro; Kawasaki, Masato; Wakatsuki, Soichi

    An increasing number of Rab GTPases associated with partial dysfunction has been linked to several human diseases characterized by a diminution in vesicle transport. Due to its direct implication in human disorders, the Rab27 subfamily is considered as a standard for vesicle docking studies. By which mechanism Rab27 effectors distinguish among the pool of Rab GTPases? What is the underneath machinery rendering the interaction of eleven distinct effectors specific of Rab27 when compared to other Rabs of the secretory pathway? By solving the X-ray structures of Rab27, both in its inactive form and active form bound to the effector protein Slp2-a, attempts have been given to unravel the molecular basis of regulation of the delivering process of vesicles to fusion by the Rab27 subfamily.

  16. Characterization of exothermic brazing components Skylab experiment M552

    NASA Technical Reports Server (NTRS)

    Pattee, H. E.; Monroe, R. E.

    1973-01-01

    Information developed to characterize flight and ground based samples from the Exothermic Brazing Experiment is detailed. Included is information developed from metallographic observation, chemical analysis, and measurements of component dimensions. Comparisons of the flight and ground based specimens showed that good quality brazes were obtained. Effects of the zero gravity processing were noted on liquid metal flow and braze alloy-base metal reactions. Unusual metallurgical structures exhibited in the nickel brazes made in Skylab were the result of composition variations apparently related to the time-temperature cycle characteristic of this braze.

  17. Small RAB GTPases Regulate Multiple Steps of Mitosis

    PubMed Central

    Miserey-Lenkei, Stéphanie; Colombo, María I.

    2016-01-01

    GTPases of the RAB family are key regulators of multiple steps of membrane trafficking. Several members of the RAB GTPase family have been implicated in mitotic progression. In this review, we will first focus on the function of endosome-associated RAB GTPases reported in early steps of mitosis, spindle pole maturation, and during cytokinesis. Second, we will discuss the role of Golgi-associated RAB GTPases at the metaphase/anaphase transition and during cytokinesis. PMID:26925400

  18. The role of the small GTPase Rab31 in cancer

    PubMed Central

    Chua, Christelle En Lin; Tang, Bor Luen

    2015-01-01

    Members of the small GTPase family Rab are emerging as potentially important factors in cancer development and progression. A good number of Rabs have been implicated or associated with various human cancers, and much recent excitement has been associated with the roles of the Rab11 subfamily member Rab25 and its effector, the Rab coupling protein (RCP), in tumourigenesis and metastasis. In this review, we focus on a Rab5 subfamily member, Rab31, and its implicated role in cancer. Well recognized as a breast cancer marker with good prognostic value, recent findings have provided some insights as to the mechanism underlying Rab31's influence on oncogenesis. Levels of Oestrogen Receptor α (ERα)- responsive Rab31 could be elevated through stabilization of its transcript by the RNA binding protein HuR, or though activation by the oncoprotein mucin1-C (MUC1-C), which forms a transcriptional complex with ERα. Elevated Rab31 stabilizes MUC1-C levels in an auto-inductive loop that could lead to aberrant signalling and gene expression associated with cancer progression. Rab31 and its guanine nucleotide exchange factor GAPex-5 have, however, also been shown to enhance early endosome-late endosome transport and degradation of the epidermal growth factor receptor (EGFR). The multifaceted action and influences of Rab31 in cancer is discussed in the light of its new interacting partners and pathways. PMID:25472813

  19. Rab24 is required for normal cell division.

    PubMed

    Militello, Rodrigo D; Munafó, Daniela B; Berón, Walter; López, Luis A; Monier, Solange; Goud, Bruno; Colombo, María I

    2013-05-01

    Rab24 is an atypical member of the Rab GTPase family whose distribution in interphase cells has been characterized; however, its function remains largely unknown. In this study, we have analyzed the distribution of Rab24 throughout cell division. We have observed that Rab24 was located at the mitotic spindle in metaphase, at the midbody during telophase and in the furrow during cytokinesis. We have also observed partial co-localization of Rab24 and tubulin and demonstrated its association to microtubules. Interestingly, more than 90% of transiently transfected HeLa cells with Rab24 presented abnormal nuclear connections (i.e., chromatin bridges). Furthermore, in CHO cells stably transfected with GFP-Rab24wt, we observed a large percentage of binucleated and multinucleated cells. In addition, these cells presented an extremely large size and multiple failures in mitosis, as aberrant spindle formation (metaphase), delayed chromosomes (telophase) and multiple cytokinesis. A marked increase in binucleated, multinucleated and multilobulated nucleus formation was observed in HeLa cells depleted of Rab24. We also present evidence that a fraction of Rab24 associates with microtubules. In addition, Rab24 knock down resulted in misalignment of chromosomes and abnormal spindle formation in metaphase leading to the appearance of delayed chromosomes during late telophase and failures in cytokinesis. Our findings suggest that an adequate level of Rab24 is necessary for normal cell division. In summary, Rab24 modulates several mitotic events, including chromosome segregation and cytokinesis, perhaps through the interaction with microtubules.

  20. 32 CFR 202.9 - Conducting RAB meetings.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... public will have a designated time on the agenda to speak to the RAB committee as a whole. (b) Nature of...) MISCELLANEOUS RESTORATION ADVISORY BOARDS Operating Requirements § 202.9 Conducting RAB meetings. (a) Public participation. RAB meetings shall be open to the public. (1) The installation co-chair shall prepare and...

  1. Simulation model of Al-Ti dissimilar laser welding-brazing and its experimental verification

    NASA Astrophysics Data System (ADS)

    Behúlová, M.; Babalová, E.; Nagy, M.

    2017-02-01

    Formation of dissimilar weld joints of light metals and alloys including Al-Ti joints is interesting mainly due to demands on the weight reduction and corrosion resistance of components and structures in automotive, aircraft, aeronautic and other industries. Joining of Al-Ti alloys represents quite difficult problem. Generally, the fusion welding of these materials can lead to the development of different metastable phases and formation of brittle intermetallic compounds. The paper deals with numerical simulation of the laser welding-brazing process of titanium Grade 2 and EN AW 5083 aluminum alloy sheets using the 5087 aluminum filler wire. Simulation model for welding-brazing of testing samples with the dimensions of 50 × 100 × 2 mm was developed in order to perform numerical experiments applying variable welding parameters and to design proper combination of these parameters for formation of sound Al-Ti welded-brazed joints. Thermal properties of welded materials in the dependence on temperature were computed using JMatPro software. The conical model of the heat source was exploited for description of the heat input to the weld due to the moving laser beam source. The sample cooling by convection and radiation to the surrounding air and shielding argon gas was taken into account. Developed simulation model was verified by comparison of obtained results of numerical simulation with the temperatures measured during real experiments of laser welding-brazing by the TruDisk 4002 disk laser.

  2. ZIRCONIUM-TITANIUM-BERYLLIUM BRAZING ALLOY

    DOEpatents

    Gilliland, R.G.; Patriarca, P.; Slaughter, G.M.; Williams, L.C.

    1962-06-12

    A new and improved ternary alloy is described which is of particular utility in braze-bonding parts made of a refractory metal selected from Group IV, V, and VI of the periodic table and alloys containing said metal as a predominating alloying ingredient. The brazing alloy contains, by weight, 40 to 50 per cent zirconium, 40 to 50 per cent titanium, and the balance beryllium in amounts ranging from 1 to 20 per cent, said alloy having a melting point in the range 950 to 1400 deg C. (AEC)

  3. Prokaryotic expression and polyclonal antibody preparation of a novel Rab-like protein mRabL5.

    PubMed

    Yang, Jie; Guo, Shi-Ying; Pan, Fei-Yan; Geng, Hui-Xia; Gong, Yi; Lou, Dan; Shu, Yong-Qian; Li, Chao-Jun

    2007-05-01

    Rab GTPases, which belong to the Ras superfamily, represent a group of small molecular weight GTP binding proteins that are involved in various steps along the exocytic and endocytic pathways. We first identified mRabL5 (GenBank Accession No. NP_080349), a novel Mus musculus Rab-like protein, present as a Golgi-associated protein. Here we presented the results of the cloning, prokaryotic expression, purification, and polyclonal antibody production of the novel Rab-like protein. In order to obtain a specific antibody against mRabL5, we prepared two GST fusion proteins, full-length mRabL5 GST fusion protein and mRabL5 C terminus GST fusion protein, to immunize rabbits. Western blot analysis showed that both antibodies prepared against full length of mRabL5 and its C terminus, respectively, can recognize mRabL5 protein. Immunofluorescence of mRabL5 in NIH3T3 cells using the two antibodies showed its perinuclear clustering distribution pattern. The polyclonal antibodies preparation against mRabL5 provided a good tool for us to study the functional involvement of mRabL5.

  4. Myosin Vc Interacts with Rab32 and Rab38 Proteins and Works in the Biogenesis and Secretion of Melanosomes*

    PubMed Central

    Bultema, Jarred J.; Boyle, Judith A.; Malenke, Parker B.; Martin, Faye E.; Dell'Angelica, Esteban C.; Cheney, Richard E.; Di Pietro, Santiago M.

    2014-01-01

    Class V myosins are actin-based motors with conserved functions in vesicle and organelle trafficking. Herein we report the discovery of a function for Myosin Vc in melanosome biogenesis as an effector of melanosome-associated Rab GTPases. We isolated Myosin Vc in a yeast two-hybrid screening for proteins that interact with Rab38, a Rab protein involved in the biogenesis of melanosomes and other lysosome-related organelles. Rab38 and its close homolog Rab32 bind to Myosin Vc but not to Myosin Va or Myosin Vb. Binding depends on residues in the switch II region of Rab32 and Rab38 and regions of the Myosin Vc coiled-coil tail domain. Myosin Vc also interacts with Rab7a and Rab8a but not with Rab11, Rab17, and Rab27. Although Myosin Vc is not particularly abundant on pigmented melanosomes, its knockdown in MNT-1 melanocytes caused defects in the trafficking of integral membrane proteins to melanosomes with substantially increased surface expression of Tyrp1, nearly complete loss of Tyrp2, and significant Vamp7 mislocalization. Knockdown of Myosin Vc in MNT-1 cells more than doubled the abundance of pigmented melanosomes but did not change the number of unpigmented melanosomes. Together the data demonstrate a novel role for Myosin Vc in melanosome biogenesis and secretion. PMID:25324551

  5. Brazing of photocathode RF gun structures in Hydrogen atmosphere: Process qualification, effect of brazing on RF properties and vacuum compatibility

    NASA Astrophysics Data System (ADS)

    Kak, Ajay; Kulshreshtha, P.; Lal, Shankar; Kaul, Rakesh; Ganesh, P.; Pant, K. K.; Abhinandan, Lala

    2012-11-01

    In this paper, we report on the development of a brazing process for an ultra-high vacuum (UHV) compatible photocathode RF gun structure developed at our Centre. The choice of brazing alloy and its form, brazing clearance between parts to be joined and the brazing cycle adopted have been qualified through metallographic examination of identical joints on an OFE copper prototype that was cut open after brazing. The quality of brazed joint not only affects the UHV compatibility of the gun, but also influences the RF parameters finally achieved. A 2-D electromagnetic code, SUPERFISH, was used to predict the variation in RF parameters before and after brazing considering actual brazing clearances provided between the parts to be joined. Results obtained from low power RF measurements on the brazed gun structure confirm the integrity of the brazed joints and show good agreement with those predicted by electromagnetic simulations. The brazed gun structure has been leak-tested and pumped down to a vacuum level limited by the vacuum compatibility of the flange-fittings employed in the setup.

  6. Active Metal Brazing and Characterization of Brazed Joints in Titanium to Carbon-Carbon Composites

    NASA Technical Reports Server (NTRS)

    Singh, M.; Shpargel, T. P.; Morscher, G. N.; Asthana, R.

    2006-01-01

    The Ti-metal/C-C composite joints were formed by reactive brazing with three commercial brazes, namely, Cu-ABA, TiCuNi, and TiCuSiI. The joint microstructures were examined using optical microscopy and scanning electron microscopy (SEM) coupled with energy dispersive spectrometry (EDS). The results of the microstructure analysis indicate solute redistribution across the joint and possible metallurgical bond formation via interdiffusion, which led to good wetting and spreading. A tube-on-plate tensile test was used to evaluate joint strength of Ti-tube/ C-C composite joints. The load-carrying ability was greatest for the Cu-ABA braze joint structures. This system appeared to have the best braze spreading which resulted in a larger braze/C-C composite bonded area compared to the other two braze materials. Also, joint loadcarrying ability was found to be higher for joint structures where the fiber tows in the outer ply of the C-C composite were aligned perpendicular to the tube axis when compared to the case where fiber tows were aligned parallel to the tube axis.

  7. Investigation of the effect of rapidly solidified braze ribbons on the microstructure of brazed joints

    NASA Astrophysics Data System (ADS)

    Bobzin, K.; Öte, M.; Wiesner, S.; Rochala, P.; Mayer, J.; Aretz, A.; Iskandar, R.; Schwedt, A.

    2017-03-01

    Shrinkage and warpage due to melting and solidification are crucial for the geometric precision of related components. In order to assure a high geometric precision, the formation of the microstructure in the joint during brazing must be taken into consideration. An extensive interaction can occur between liquid melt and base material, resulting in the formation of distinctive phases. This interaction depends on the parameters of the brazing process. However, the consequences of the interaction between phase formation and process parameters in terms of geometric precision cannot be estimated yet. Insufficient quality of the joint can be a result. In this study, investigations focus on the process of solidification in terms of time dependent diffusion behavior of elements. Therefore, microcrystalline and amorphous braze ribbons based on Ti are produced by rapid solidification and are used for joining. The microstructure of the braze ribbons as well as the melting behavior and phase formation during brazing are considered to be of particular importance for the mechanical properties of the brazed components.

  8. Rabifier2: an improved bioinformatic classifier of Rab GTPases.

    PubMed

    Surkont, Jaroslaw; Diekmann, Yoan; Pereira-Leal, José B

    2016-10-22

    The Rab family of small GTPases regulates and provides specificity to the endomembrane trafficking system; each Rab subfamily is associated with specific pathways. Thus, characterization of Rab repertoires provides functional information about organisms and evolution of the eukaryotic cell. Yet, the complex structure of the Rab family limits the application of existing methods for protein classification. Here, we present a major redesign of the Rabifier, a bioinformatic pipeline for detection and classification of Rab GTPases. It is more accurate, significantly faster than the original version and is now open source, both the code and the data, allowing for community participation.

  9. Thermal and Fluid Flow Brazing Simulations

    SciTech Connect

    HOSKING, FLOYD MICHAEL; GIANOULAKIS,STEVEN E.; GIVLER,RICHARD C.; SCHUNK,P. RANDALL

    1999-12-15

    The thermal response of fixtured parts in a batch-type brazing furnace can require numerous, time-consuming development runs before an acceptable furnace schedule or joint design is established. Powerful computational simulation tools are being developed to minimize the required number of verification experiments, improve furnace throughput, and increase product yields. Typical furnace simulations are based on thermal, fluid flow, and structural codes that incorporate the fundamental physics of the brazing process. The use of massively parallel computing to predict furnace and joint-level responses is presented. Measured and computed data are compared. Temperature values are within 1-270 of the expected peak brazing temperature for different loading conditions. Sensitivity studies reveal that the thermal response is more sensitive to the thermal boundary conditions of the heating enclosure than variability y in the materials data. Braze flow simulations predict fillet geometry and free surface joint defects. Dynamic wetting conditions, interfacial reactions, and solidification structure add a high degree of uncertainty to the flow results.

  10. Diffusion brazing nickel-plated stainless steel

    NASA Technical Reports Server (NTRS)

    Beuyukian, C. S.; Mitchell, M. J.

    1976-01-01

    To bond parts, sandwich assembly is made up of aluminum core, aluminum face sheet with brazing alloy interface, and nickel plated stainless steel part. Sandwich is placed between bottom and top glide sheet that is placed in stainless steel retort where assembly is bonded at 580 C.

  11. Welding/brazing for Space Station repair

    NASA Technical Reports Server (NTRS)

    Dickinson, David W.; Babel, H. W.; Conaway, H. R.; Hooper, W. H.

    1990-01-01

    Viewgraphs on welding/brazing for space station repair are presented. Topics covered include: fabrication and repair candidates; debris penetration of module panel; welded repair patch; mechanical assembly of utility fluid line; space station utility systems; Soviet aerospace fabrication - an overview; and processes under consideration.

  12. Nickel-chromium-silicon brazing filler metal

    DOEpatents

    Martini, Angelo J.; Gourley, Bruce R.

    1976-01-01

    A brazing filler metal containing, by weight percent, 23-35% chromium, 9-12% silicon, a maximum of 0.15% carbon, and the remainder nickel. The maximum amount of elements other than those noted above is 1.00%.

  13. Gold-nickel-titanium brazing alloy

    DOEpatents

    Mizuhara, Howard

    1990-07-03

    A brazing alloy in accordance with this invention has the following composition, by weight: 91 to 99% gold, 0.5 to 7% nickel; 0.10 to 2% titanium. Alternatively, with palladium present, the composition is as follows, by weight: 83 to 96% gold; 3 to 10% palladium; 0.5 to 5% nickel; 0.10 to 2% titanium.

  14. Brazed bipolar plates for PEM fuel cells

    DOEpatents

    Neutzler, Jay Kevin

    1998-01-01

    A liquid-cooled, bipolar plate separating adjacent cells of a PEM fuel cell comprising corrosion-resistant metal sheets brazed together so as to provide a passage between the sheets through which a dielectric coolant flows. The brazement comprises a metal which is substantially insoluble in the coolant.

  15. Brazed bipolar plates for PEM fuel cells

    DOEpatents

    Neutzler, J.K.

    1998-07-07

    A liquid-cooled, bipolar plate separating adjacent cells of a PEM fuel cell comprises corrosion-resistant metal sheets brazed together so as to provide a passage between the sheets through which a dielectric coolant flows. The brazement comprises a metal which is substantially insoluble in the coolant. 6 figs.

  16. Gold-nickel-titanium brazing alloy

    DOEpatents

    Mizuhara, Howard

    1995-01-03

    A brazing alloy in accordance with this invention has the following composition, by weight: 91 to 99 gold, 0.5 to 7% nickel; 0.10 to 2% titanium. Alternatively, with palladium present, the composition is as follows, by weight: 83 to 96% gold; 3 to 10% palladium; 0.5 to 5% nickel; 0.10 to 2% titanium.

  17. Crystal structure of inactive form of Rab3B

    SciTech Connect

    Zhang, Wei; Shen, Yang; Jiao, Ronghong; Liu, Yanli; Deng, Lingfu; Qi, Chao

    2012-02-24

    Highlights: Black-Right-Pointing-Pointer This is the first structural information of human Rab3B. Black-Right-Pointing-Pointer To provides a structural basis for the GDP/GTP switch in controlling the activity of Rab3. Black-Right-Pointing-Pointer The charge distribution of Rab3B indicates its unique roles in vesicular trafficking. -- Abstract: Rab proteins are the largest family of ras-related GTPases in eukaryotic cells. They act as directional molecular switches at membrane trafficking, including vesicle budding, cargo sorting, transport, tethering, and fusion. Here, we generated and crystallized the Rab3B:GDP complex. The structure of the complex was solved to 1.9 A resolution and the structural base comparison with other Rab3 members provides a structural basis for the GDP/GTP switch in controlling the activity of small GTPase. The comparison of charge distribution among the members of Rab3 also indicates their different roles in vesicular trafficking.

  18. Multiple Roles of the Small GTPase Rab7

    PubMed Central

    Guerra, Flora; Bucci, Cecilia

    2016-01-01

    Rab7 is a small GTPase that belongs to the Rab family and controls transport to late endocytic compartments such as late endosomes and lysosomes. The mechanism of action of Rab7 in the late endocytic pathway has been extensively studied. Rab7 is fundamental for lysosomal biogenesis, positioning and functions, and for trafficking and degradation of several signaling receptors, thus also having implications on signal transduction. Several Rab7 interacting proteins have being identified leading to the discovery of a number of different important functions, beside its established role in endocytosis. Furthermore, Rab7 has specific functions in neurons. This review highlights and discusses the role and the importance of Rab7 on different cellular pathways and processes. PMID:27548222

  19. Vacuum brazing of carbon nanotube strands

    NASA Astrophysics Data System (ADS)

    Wu, Wei

    Carbon nanotubes (CNTs) discovered at 1991 have attracted great interest for applications in Nano-Electro-Mechanical-Systems (NEMS). However, the search for methods to join CNTs with metallic parts has been a worldwide challenge. Many efforts have been devoted to manipulating individual CNTs and joining them to each other. Joining processes so far attempted are premature and fall short of efficiency for joint quality evaluation. Thus, it has been found necessary to work on macro CNTs strands which are easy to handle via macro joining techniques. In this study, vacuum brazing technology has been developed for joining macro CNTs strands with Ni using a Ti-Ag-Cu alloy. The brazing mechanism has been confirmed as due to TiC formation at the CNTs/Ti-Ag-Cu interface. To evaluate this novel vacuum brazing technique for CNTs joining, the temperature effect on the brazing mechanism, microstructure and stoichiometry at joint interface needed to be understood. Firstly, the influence of temperature (from room temperature to 1000°C) on mechanical behaviour of CNTs was well examined. The ultimate tensile strength (UTS) of CNTs was measured to be a maximum at 900°C. Then, the mechanical performance of the joints was investigated from 850°C to 1000°C, and the fracture modes of the joints were identified. The UTS of joint also achieves maximum at 900°C. Below 900°C, due to little TiC formation, the bonding is weak thus leading to interfacial fracture. Above 900°C, due to much TiC formation, the bonding is strong thus resulting in CNTs fracture. Furthermore, the vacuum brazing technique was applied to join CNTs to Ni contact wires used as a lamp filament. Compared to the filament joined by Ag paste or mechanical connection, the illumination of the brazed CNTs filament was stronger. The current density of the brazed filament was superior to the Ag paste connected filament. This may represent a promising way to produce energy saving lamps.

  20. Development of the weld-braze joining process

    NASA Technical Reports Server (NTRS)

    Bales, T. T.; Royster, D. M.; Arnold, W. E., Jr.

    1973-01-01

    A joining process, designated weld-brazing, was developed which combines resistance spot welding and brazing. Resistance spot welding is used to position and aline the parts, as well as to establish a suitable faying-surface gap for brazing. Fabrication is then completed at elevated temperature by capillary flow of the braze alloy into the joint. The process was used successfully to fabricate Ti-6Al-4V alloy joints by using 3003 aluminum braze alloy and should be applicable to other metal-braze systems. Test results obtained on single-overlap and hat-stiffened panel specimens show that weld-brazed joints were superior in tensile shear, stress rupture, fatigue, and buckling compared with joints fabricated by conventional means. Another attractive feature of the process is that the brazed joint is hermetically sealed by the braze material, which may eliminate many of the sealing problems encountered with riveted or spot welded structures. The relative ease of fabrication associated with the weld-brazing process may make it cost effective over conventional joining techniques.

  1. [Rab GTPases networks in membrane traffic in Saccharomyces cerevisiae].

    PubMed

    Nagano, Makoto; Toshima, Junko Y; Toshima, Jiro

    2015-01-01

    Intracellular membrane trafficking between membranous compartments is essential for organelle biogenesis, structure, and identity. Rab/Ypt GTPases are well-characterized regulators of intracellular membrane trafficking, functioning as molecular switches that alternate between GTP- and GDP-bound forms. In Saccharomyces cerevisiae, 11 Rab/Ypt GTPases have been identified and their functions are known to be conserved in their mammalian counterparts. In yeast, the secretory pathway is regulated by sequential activation and inactivation (the so-called Rab cascade) of three types of yeast Rab protein -Ypt1p, Ypt31p/32p and Sec4p -via specific guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). In addition to these Rabs, we and others have recently demonstrated that Ypt6p is predominantly localized to the early Golgi compartment, and functions as another regulator of anterograde transport for intra-Golgi trafficking in the secretory pathway. On the other hand, the endocytic pathway is known to be regulated by three yeast Rab5s (Vps21p, Ypt52p and Ypt53p) and one Rab7 (Ypt7p). Rab5 and Rab7 are key determinants of endosome identity, and the Rab5-Rab7 cascade is important for the progression from early to late endosome. Our recent study demonstrates that the endocytic pathway branches into two vacuolar targeting pathways, the Rab5-dependent vacuole protein sorting (VPS) pathway and the Rab5-independent pathway. In this review, we focus on recent advances in our understanding of molecular mechanisms that regulate the localization and activity of yeast Rab GTPases in intracellular membrane trafficking.

  2. Rab 7: an important regulator of late endocytic membrane traffic

    PubMed Central

    1995-01-01

    Rab5 and rab7 proteins belong to a superfamily of small molecular weight GTPases known to be associated with early and late endosomes, respectively. The rab5 protein plays an important regulatory role in early endocytosis, yet the function of rab7 protein was previously uncharacterized. This question was addressed by comparing the kinetics of vesicular stomatitis virus (VSV) G protein internalization in baby hamster kidney cells overexpressing wild-type or dominant negative mutant forms of the rab7 protein (rab7N125I and rab7T22N). Overexpression of wild-type rab7 protein allowed normal transport to late endosomes (mannose 6-phosphate receptor positive), while the rab7N125I mutant caused the VSV G protein to accumulate specifically in early (transferrin receptor positive) endosomes. Horseradish peroxidase and paramyxovirus SV5 hemagglutinin-neuraminidase (HN) were used in quantitative biochemical assays to further demonstrate that rab7 function was not required for early internalization events, but was crucial in downstream degradative events. The characteristic cleavage of SV5 HN in the late endosome distinguishes internalization from transport to later stages of the endocytic pathway. Mutant rab7N125I or rab7T22N proteins had no effect on the internalization of either horseradish peroxidase or SV5 HN protein. In contrast, the mutant proteins markedly inhibited the subsequent cleavage of the SV5 HN protein. Taken together, these data support a key role for rab7, downstream of rab5, in regulating membrane transport leading from early to late endosomes. We compare our findings to those obtained for the yeast homologues Ypt51p, Ypt52p, Ypt53p, and Ypt7p. PMID:8522602

  3. Rab8a/Rab11a regulate intercellular communications between neural cells via tunneling nanotubes

    PubMed Central

    Zhu, Hui; Xue, Chengbin; Xu, Xi; Guo, Yibing; Li, Xiaohong; Lu, Jingjing; Ju, Shaoqing; Wang, Yongjun; Cao, Zheng; Gu, Xiaosong

    2016-01-01

    Tunneling nanotubes (TNTs) are F-actin-based membrane tubes, and can form between cultured cells and within vital tissues. TNTs mediate intercellular communications that range from electrical signaling to the transfer of organelles. Following peripheral nerve injury, the orchestrated intercellular communications among neural and non-neural cells are required for effective nerve regeneration. It remains unknown whether TNTs exist between neural cells in the peripheral nerve system and how TNTs affect neural regeneration. To address these interesting questions, we investigated the transfer of neurotropic factors, membrane protein, cytoplasmic protein, mitochondria and RNA in functional TNTs formed between cultured Schwann cells (SCs). TNT-like structures were increased not only in cultured SCs after exposure to serum depletion but also in longitudinal sections of proximal sciatic nerve stump harvested after rat peripheral nerve transection. Meanwhile, downregulation of Rab8a or Rab11a in cultured SCs inhibited the formation of functional TNTs and vesicle transfer and led to decrease in cell migration, increase in SCs apoptosis. Likewise, knockdown of Rab8a or Rab11a in primary SCs also suppressed axonal outgrowth from co-cultured dorsal root ganglion (DRG) neurons. Overall, our results suggested that the gene of Rab8a or Rab11a might be involved in the formation of TNTs structures in the peripheral nerve system, while TNTs structures were likely to affect peripheral nerve regeneration through the regulation of neural cell communications. PMID:28005071

  4. Several braze filler metals for joining an oxide-dispersion-strengthened nickel-chromium-aluminum alloy

    NASA Technical Reports Server (NTRS)

    Gyorgak, C. A.

    1975-01-01

    An evaluation was made of five braze filler metals for joining an aluminum-containing oxide dispersion-strengthened (ODS) alloy, TD-NiCrAl. All five braze filler metals evaluated are considered suitable for joining TD-NiCrAl in terms of wettability and flow. Also, the braze alloys appear to be tolerant of slight variations in brazing procedures since joints prepared by three sources using three of the braze filler metals exhibited similar brazing characteristics and essentially equivalent 1100 C stress-rupture properties in a brazed butt-joint configuration. Recommendations are provided for brazing the aluminum-containing ODS alloys.

  5. Characterization of a RAB5 homologue in Trypanosoma cruzi.

    PubMed

    Araripe, Júlia Rolão; Ramos, Fabiane Pereira; Cunha e Silva, Narcisa Leal; Urményi, Turán Péter; Silva, Rosane; Leite Fontes, Carlos Frederico; da Silveira, José Franco; Rondinelli, Edson

    2005-04-08

    RAB proteins are small GTPases involved in exocytic and endocytic pathways of eukaryotic cells, controlling vesicle docking and fusion. RABs show a remarkable specificity in subcellular localization, so they can be used as molecular markers for studying protein trafficking in Trypanosoma cruzi, the causal agent of Chagas' disease. RAB5 is a component of early endosomes. It has been identified in kinetoplastids such as Trypanosoma brucei and Leishmania donovani. In this work, we describe the characterization of the complete coding sequence of a RAB5 gene homologue in T. cruzi (TcRAB5, GenBank Accession No. AY730667). It is present as a single copy gene, located at chromosomal bands XIII and XIV. TcRAB5 shares the highest degrees of similarity (71%) and identity (63%) with Trypanosoma brucei rhodesiense RAB5a and contains all five characteristic RAB motifs. TcRAB5 is transcribed as a single 1.5kb mRNA in epimastigotes. Its transcript was also detected in the other two forms of the parasite, metacyclic trypomastigotes and spheromastigotes. The recombinant TcRAB5 protein was able to bind and hydrolyze GTP. The identification of proteins involved in T. cruzi endo- and exocytic pathways may generate cellular compartment markers, an invaluable tool to better understand the vesicular transport in this parasite.

  6. Mechanical properties of high-temperature brazed titanium materials

    SciTech Connect

    Lugscheider, E.; Broich, U.; Koetzing, B.

    1994-12-31

    Titanium and its alloys are of main interest for several fields of application. Because of rising demands on permanent structural parts and increasing complexity of components, it is important to obtain an adequate joining technique, which on the one hand does not restrict the mechanical properties of the parent metal too much and on the other hand is inexpensive and flexible. These requirements can be fulfilled best by application of high-temperature brazing technology, employing titanium base filler metals. Among existing joining techniques, the vacuum brazing process conducted in a vacuum furnace and the induction brazing process are of practical relevance. The mechanical properties of high-temperature brazed titanium materials are strongly dependent on process parameters, such as brazing time and brazing temperature and even more crucially on brazing gap size. Under optimized brazing conditions, the tensile strength of high-temperature brazed TiAl6V4-joints, for example, reach about 950 MPa, where the Pd-containing alloy is slightly superior to TiCu20Ni20 filler metal. Most of the tensile specimens break in the base metal at some distance to the brazing zone, indicating that the tensile strength of the joint is comparable to that of the bulk material. The thermal stability of the brazed titanium joints has been investigated by doing tensile tests at elevated temperatures. These experiments have shown that the tensile strength of TiAl6VA- joints for example are going to be reduced by 30% when exposing the brazed samples to 300{degrees}C. However, the tensile specimen broke in the bulk material, indicating that the tensile strength of bulk TiAl6V4 is also reduced at elevated temperatures.

  7. Applying Taguchi Methods To Brazing Of Rocket-Nozzle Tubes

    NASA Technical Reports Server (NTRS)

    Gilbert, Jeffrey L.; Bellows, William J.; Deily, David C.; Brennan, Alex; Somerville, John G.

    1995-01-01

    Report describes experimental study in which Taguchi Methods applied with view toward improving brazing of coolant tubes in nozzle of main engine of space shuttle. Dr. Taguchi's parameter design technique used to define proposed modifications of brazing process reducing manufacturing time and cost by reducing number of furnace brazing cycles and number of tube-gap inspections needed to achieve desired small gaps between tubes.

  8. Evaluation of Brazed Joints Using Failure Assessment Diagram

    NASA Technical Reports Server (NTRS)

    Flom, Yury

    2012-01-01

    Fitness-for service approach was used to perform structural analysis of the brazed joints consisting of several base metal / filler metal combinations. Failure Assessment Diagrams (FADs) based on tensile and shear stress ratios were constructed and experimentally validated. It was shown that such FADs can provide a conservative estimate of safe combinations of stresses in the brazed joints. Based on this approach, Margins of Safety (MS) of the brazed joints subjected to multi-axial loading conditions can be evaluated..

  9. Thermal fatigue and oxidation data for alloy/braze combinations

    NASA Technical Reports Server (NTRS)

    Hill, V. L.; Humphreys, V. E.

    1977-01-01

    Thermal fatigue and oxidation data were obtained for 62 brazed specimens of 3 iron-, 3 nickel-, and 1 cobalt-base alloy. Fluidized bed thermal cycling was conducted over the range 740/25 C employing 10 cm long single-edge wedge specimens. Immersion time was always 4 minutes in each bed. Types of test specimens employed in the program include those with brazed overlays on the specimen radius, those butt brazed at midspan and those with a brazed foil overlay on the specimen radius. Of the 18 braze overlay specimens, 5 generated fatigue cracks by 7000 cycles. Thermal cracking of butt brazed specimens occurred exclusively through the butt braze. Of the 23 butt brazed specimens, 7 survived 11,000 thermal cycles without cracking. Only 2 of the 21 foil overlaid specimens exhibiting cracking in 7,000 cycles. Blistering of the foil did occur for 2 alloys by 500 cycles. Oxidation of the alloy/braze combination was limited at the test maximum test temperature of 740 C.

  10. Ultrasonic scanning system for inspection of brazed tube joints

    NASA Technical Reports Server (NTRS)

    Haynes, J. L.; Maurer, N. A.

    1972-01-01

    An ultrasonic scanning system used to inspect and evaluate in-place brazed tube joints is considered. The system was designed, developed, and built especially for nondestructive testing and was selected because of its known response to brazing defects not associated with material density changes. The scan system is capable of scanning brazed joints in union, tee, elbow and cross configuration of 3/16-inch through 5/8-inch diameters. It is capable of detecting brazed defects as small as 0.008 by 0.010-inch, which exceeds the 0.015-inch diameter defect resolution required by specification.

  11. Spot brazing of aluminum to copper with a cover plate

    NASA Astrophysics Data System (ADS)

    Hayashi, Junya; Miyazawa, Yasuyuki

    2014-08-01

    It is difficult to join dissimilar metals when an intermetallic compound is formed at the joining interface. Spot brazing can be accomplished in a short time by resistance heating. Therefore, it is said that the formation of a intermetallic compound can be prevented. In this study, aluminum and copper were joined by spot brazing with a cover plate. The cover plate was used to supply heat to base metals and prevent heat dissipation from the base metals. The ability to braze Al and Cu was investigated by observation and analysis. Pure aluminum (A1050) plate and oxygen-free copper (C1020) plate were used as base metals. Cu-Ni-Sn-P brazing filler was used as the brazing filler metal. SPCC was employed as cover plate. Brazing was done with a micro spot welder under an argon gas atmosphere. Brazing ability was estimated by tensile shear strength and cross sectional microstructure observation. Al and Cu can be joined by spot brazing with Cu-Ni-Sn-P brazing filler and cover plate.

  12. Composition and method for brazing graphite to graphite

    DOEpatents

    Taylor, Albert J.; Dykes, Norman L.

    1984-01-01

    The present invention is directed to a brazing material for joining graphite structures that can be used at temperatures up to about 2800.degree. C. The brazing material formed of a paste-like composition of hafnium carbide and uranium oxide with a thermosetting resin. The uranium oxide is converted to uranium dicarbide during the brazing operation and then the hafnium carbide and uranium dicarbide form a liquid phase at a temperature about 2600.degree. C. with the uranium diffusing and vaporizing from the joint area as the temperature is increased to about 2800.degree. C. so as to provide a brazed joint consisting essentially of hafnium carbide. This brazing temperature for hafnium carbide is considerably less than the eutectic temperature of hafnium carbide of about 3150.degree. C. The brazing composition also incorporates the thermosetting resin so that during the brazing operation the graphite structures may be temporarily bonded together by thermosetting the resin so that machining of the structures to final dimensions may be completed prior to the completion of the brazing operation. The resulting brazed joint is chemically and thermally compatible with the graphite structures joined thereby and also provides a joint of sufficient integrity so as to at least correspond with the strength and other properties of the graphite.

  13. A family of Rab27-binding proteins. Melanophilin links Rab27a and myosin Va function in melanosome transport.

    PubMed

    Strom, Molly; Hume, Alistair N; Tarafder, Abul K; Barkagianni, Eleni; Seabra, Miguel C

    2002-07-12

    The Rab27a GTPase regulates diverse processes involving lysosome-related organelles, including melanosome motility in melanocytes, and lytic granule release in cytotoxic T lymphocytes. Toward an understanding of Rab27a function, we searched for proteins that interact with Rab27a(GTP) using the yeast two-hybrid system and identified JFC1/Slp1, a protein of unknown function. JFC1/Slp1 and related proteins, including melanophilin, contain a conserved amino-terminal domain similar to the Rab3a-binding domain of Rabphilin-3. We used several methods to demonstrate that this conserved amino-terminal domain is a Rab27-binding domain. We show that this domain interacts directly, and in a GTP-dependent manner with Rab27a. Furthermore, overexpression of this domain in melanocytes results in perinuclear clustering of melanosomes, suggesting that this region is sufficient for interaction with, and perturbation of function of, Rab27a in a physiological context. Thus, we identified a novel family of Rab27-binding proteins. We also show that melanophilin associates with Rab27a and myosin Va on melanosomes in melanocytes, and present evidence that a domain within the carboxyl-terminal region of melanophilin interacts with the carboxyl-terminal tail of the melanocyte-specific splice isoform of myosin Va. Thus, melanophilin can associate simultaneously with activated Rab27a and myosin Va via distinct regions, and serve as a linker between these proteins.

  14. Brazing as a Means of Sealing Ceramic Membranes for use in Advanced Coal Gasification Processes

    SciTech Connect

    Weil, K. Scott; Hardy, John S.; Rice, Joseph P.; Kim, Jin Yong Y.

    2006-01-02

    Coal is potentially a very inexpensive source of clean hydrogen fuel for use in fuel cells, turbines, and various process applications. To realize its potential however, efficient, low-cost gas separation systems are needed to provide high purity oxygen to enhance the coal gasification reaction and to extract hydrogen from the resulting gas product stream. Several types of inorganic membranes are being developed for hydrogen or oxygen separation, including porous alumina, transition metal oxide perovskites, and zirconia. One of the key challenges in developing solid-state membrane based gas separation systems is in hermetically joining the membrane to the metallic body of the separation device. In an effort to begin addressing this issue, a new brazing concept has been developed, referred to as reactive air brazing. This paper discusses the details of this joining technique and illustrates its use in bonding a wide variety of materials, including alumina, lanthanum strontium cobalt ferrite, and yttria stabilized zirconia.

  15. The story of laser brazing technology

    NASA Astrophysics Data System (ADS)

    Hoffmann, Peter; Dierken, Roland

    2012-03-01

    This article gives an overview on the development of laser brazing technology as a new joining technique for car body production. The story starts with fundamental research work at German institutes in 1993, continues with the first implementations in automobile production in 1998, gives examples of applications since then and ends with an outlook. Laser brazing adapted design of joints and boundary conditions for a safe processing are discussed. Besides a better understanding for the sensitivity of the process against joint irregularities and misalignment, the key to successful launch was an advanced system technology. Different working heads equipped with wire feeding device, seam tracking system or tactile sensors for an automated teaching are presented in this paper. Novel laser heads providing a two beam technology will allow improved penetration depth of the filler wire and a more ecological processing by means of energy consumption.

  16. A viscoplastic theory for braze alloys

    SciTech Connect

    Neilsen, M.K.; Burchett, S.N.; Stone, C.M.; Stephens, J.J.

    1996-04-01

    A new viscoplastic theory for CusilABA and other braze alloys has been developed. Like previous viscoplastic theories,this new theory uses a hyperbolic sine function of effective stress in its kinetic equation for the inelastic strain rate. This new theory has an internal state variable which accounts for isotropic hardening and recovery and a second-order, internal state tensor which accounts for kinematic hardening and recovery. Unlike previous theories, the new theory uses evolution equations for the state variables which describe competing mechanisms of power law hardening and static recovery. The evolution equations used in previous theories describe competing mechanisms of linear hardening, dynamic and static recovery. The new viscoplastic theory was implemented in several finite element codes and used in several metal-to-ceramic brazing simulations. Two approaches for obtaining material parameters for the new viscoplastic theory were developed.

  17. An Ag based brazing system with a tunable thermal expansion for the use as sealant for solid oxide cells

    NASA Astrophysics Data System (ADS)

    Kiebach, Ragnar; Engelbrecht, Kurt; Grahl-Madsen, Laila; Sieborg, Bertil; Chen, Ming; Hjelm, Johan; Norrman, Kion; Chatzichristodoulou, Christodoulos; Hendriksen, Peter Vang

    2016-05-01

    An Ag-Al2TiO5 composite braze was developed and successfully tested as seal for solid oxide cells. The thermo-mechanical properties of the Ag-Al2TiO5 system and the chemical compatibility between this composite braze and relevant materials used in stacks were characterized and the leak rates as a function of the operation temperature were measured. The thermal expansion coefficient in the Ag-Al2TiO5 system can be tailored by varying the amount of the ceramic filler. The brazing process can be carried out in air, the joining partners showed a good chemical stability and sufficient low leak rates were demonstrated. Furthermore, the long-term stability of the Ag-Al2TiO5 composite braze was studied under relevant SOFC and SOEC conditions. The stability of brazed Crofer/Ag-Al2TiO5/NiO-YSZ assemblies in reducing atmosphere and in pure oxygen was investigated over 500 h at 850 °C. Additionally, a cell component test was performed to investigate the durability of the Ag-Al2TiO5 seal when exposed to dual atmosphere. The seals performed well over 900 h under electrolysis operation conditions (-0.5 A cm2, 850 °C), and no cell degradation related to the Ag-Al2TiO5 sealing was found, indicating that the developed braze system is applicable for the use in SOFC/SOEC stacks.

  18. Brazing of copper to stainless steel with a low-silver-content brazing filler metal

    NASA Astrophysics Data System (ADS)

    Fukikoshi, Tatsuya; Watanabe, Yūki; Miyazawa, Yasuyuki; Kanasaki, Fumio

    2014-08-01

    The brazing of copper to stainless steel (SUS304 JIS) was performed using a low- silver-content brazing filler metal, Ag-50Cu, under an Ar gas atmosphere with a conventional furnace, owing to the potential economic benefits of using low-silver-content filler metals. The brazeability of the low-silver-content brazing filler metal to copper and SUS304 was investigated. A good joint was obtained, and a drastic dissolution reaction occurred at the copper side. Molten BAg8 penetrated along the crystal grain boundary of the copper base metal when BAg8 was used as the filler metal. This was caused by the dissolution of Ni from the stainless steel into the molten filler metal. Ag-50Cu, which was investigated in this work, can be used instead of BAg8 filler metal.

  19. Phosphonocarboxylates Inhibit the Second Geranylgeranyl Addition by Rab Geranylgeranyl Transferase*

    PubMed Central

    Baron, Rudi A.; Tavaré, Richard; Figueiredo, Ana C.; Błażewska, Katarzyna M.; Kashemirov, Boris A.; McKenna, Charles E.; Ebetino, Frank H.; Taylor, Adam; Rogers, Michael J.; Coxon, Fraser P.; Seabra, Miguel C.

    2009-01-01

    Rab geranylgeranyl transferase (RGGT) catalyzes the post-translational geranylgeranyl (GG) modification of (usually) two C-terminal cysteines in Rab GTPases. Here we studied the mechanism of the Rab geranylgeranylation reaction by bisphosphonate analogs in which one phosphonate group is replaced by a carboxylate (phosphonocarboxylate, PC). The phosphonocarboxylates used were 3-PEHPC, which was previously reported, and 2-hydroxy-3-imidazo[1,2-a]pyridin-3-yl-2-phosphonopropionic acid ((+)-3-IPEHPC), a >25-fold more potent related compound as measured by both IC50 and Ki.(+)-3-IPEHPC behaves as a mixed-type inhibitor with respect to GG pyrophosphate (GGPP) and an uncompetitive inhibitor with respect to Rab substrates. We propose that phosphonocarboxylates prevent only the second GG transfer onto Rabs based on the following evidence. First, geranylgeranylation of Rab proteins ending with a single cysteine motif such as CAAX, is not affected by the inhibitors, either in vitro or in vivo. Second, the addition of an -AAX sequence onto Rab-CC proteins protects the substrate from inhibition by the inhibitors. Third, we demonstrate directly that in the presence of (+)-3-IPEHPC, Rab-CC and Rab-CXC proteins are modified by only a single GG addition. The presence of (+)-3-IPEHPC resulted in a preference for the Rab N-terminal cysteine to be modified first, suggesting an order of cysteine geranylgeranylation in RGGT catalysis. Our results further suggest that the inhibitor binds to a site distinct from the GGPP-binding site on RGGT. We suggest that phosphonocarboxylate inhibitors bind to a GG-cysteine binding site adjacent to the active site, which is necessary to align the mono-GG-Rab for the second GG addition. These inhibitors may represent a novel therapeutic approach in Rab-mediated diseases. PMID:19074143

  20. Copper-phosphorus alloys offer advantages in brazing copper

    SciTech Connect

    Rupert, W.D.

    1996-05-01

    Copper-phosphorus brazing alloys are used extensively for joining copper, especially refrigeration and air-conditioning copper tubing and electrical conductors. What is the effect of phosphorus when alloyed with copper? The following are some of the major effects: (1) It lowers the melt temperature of copper (a temperature depressant). (2) It increases the fluidity of the copper when in the liquid state. (3) It acts as a deoxidant or a fluxing agent with copper. (4) It lowers the ductility of copper (embrittles). There is a misconception that silver improves the ductility of the copper-phosphorus alloys. In reality, silver added to copper acts in a similar manner as phosphorus. The addition of silver to copper lowers the melt temperature (temperature depressant) and decreases the ductility. Fortunately, the rate and amount at which silver lowers copper ductility is significantly less than that of phosphorus. Therefore, taking advantage of the temperature depressant property of silver, a Ag-Cu-P alloy can be selected at approximately the same melt temperature as a Cu-P alloy, but at a lower phosphorus content. The lowering of the phosphorus content actually makes the alloy more ductile, not the silver addition. A major advantage of the copper-phosphorus alloys is the self-fluxing characteristic when joining copper to copper. They may also be used with the addition of a paste flux on brass, bronze, and specialized applications on silver, tungsten and molybdenum. Whether it is selection of the proper BCuP alloy or troubleshooting an existing problem, the suggested approach is a review of the desired phosphorus content in the liquid metal and how it is being altered during application. In torch brazing, a slight change in the oxygen-fuel ratio can affect the joint quality or leak tightness.

  1. Elevated temperature creep properties for selected active metal braze alloys

    SciTech Connect

    Stephens, J.J.

    1997-02-01

    Active metal braze alloys reduce the number of processes required for the joining of metal to ceramic components by eliminating the need for metallization and/or Ni plating of the ceramic surfaces. Titanium (Ti), V, and Zr are examples of active element additions which have been used successfully in such braze alloys. Since the braze alloy is expected to accommodate thermal expansion mismatch strains between the metal and ceramic materials, a knowledge of its elevated temperature mechanical properties is important. In particular, the issue of whether or not the creep strength of an active metal braze alloy is increased or decreased relative to its non-activated counterpart is important when designing new brazing processes and alloy systems. This paper presents a survey of high temperature mechanical properties for two pairs of conventional braze alloys and their active metal counterparts: (a) the conventional 72Ag-28Cu (Cusil) alloy, and the active braze alloy 62.2Ag- 36.2Cu-1.6Ti (Cusil ABA), and (b) the 82Au-18Ni (Nioro) alloy and the active braze alloy Mu-15.5M-0.75Mo-1.75V (Nioro ABA). For the case of the Cusil/Cusil ABA pair, the active metal addition contributes to solid solution strengthening of the braze alloy, resulting in a higher creep strength as compared to the non-active alloy. In the case of the Nioro/Nioro ABA pair, the Mo and V additions cause the active braze alloy to have a two-phase microstructure, which results in a reduced creep strength than the conventional braze alloy. The Garofalo sinh equation has been used to quantitatively describe the stress and temperature dependence of the deformation behavior. It will be observed that the effective stress exponent in the Garofalo sinh equation is a function of the instantaneous value of the stress argument.

  2. Rab5 Isoforms Orchestrate a “Division of Labor” in the Endocytic Network; Rab5C Modulates Rac-Mediated Cell Motility

    PubMed Central

    Chen, Pin-I; Schauer, Kristine; Kong, Chen; Harding, Andrew R.; Goud, Bruno; Stahl, Philip D.

    2014-01-01

    Rab5, the prototypical Rab GTPase and master regulator of the endocytic pathway, is encoded as three differentially expressed isoforms, Rab5A, Rab5B and Rab5C. Here, we examined the differential effects of Rab5 isoform silencing on cell motility and report that Rab5C, but neither Rab5A nor Rab5B, is selectively associated with the growth factor-activation of Rac1 and with enhanced cell motility. Initial observations revealed that silencing of Rab5C expression, but neither Rab5A nor Rab5C, led to spindle-shaped cells that displayed reduced formation of membrane ruffles. When subjected to a scratch wound assay, cells depleted of Rab5C, but not Rab5A or Rab5B, demonstrated reduced cell migration. U937 cells depleted of Rab5C also displayed reduced cell motility in a Transwell plate migration assay. To examine activation of Rac, HeLa cells stably expressing GFP-Rac1 were independently depleted of Rab5A, Rab5B or Rab5C and seeded onto coverslips imprinted with a crossbow pattern. 3-D GFP-Rac1 images of micro-patterned cells show that GFP-Rac1 was less localized to the cell periphery in the absence of Rab5C. To confirm the connection between Rab5C and Rac activation, HeLa cells depleted of Rab5 isoforms were starved and then stimulated with EGF. Rac1 pull-down assays revealed that EGF-stimulated Rac1 activity was significantly suppressed in Rab5C-suppressed cells. To determine whether events upstream of Rac activation were affected by Rab5C, we observed that EGF-stimulated Akt phosphorylation was suppressed in cells depleted of Rab5C. Finally, since spatio-temporal assembly/disassembly of adhesion complexes are essential components of cell migration, we examined the effect of Rab5 isoform depletion on the formation of focal adhesion complexes. Rab5C-depleted HeLa cells have significantly fewer focal adhesion foci, in accordance with the lack of persistent lamellipodial protrusions and reduced directional migration. We conclude that Rab5 isoforms selectively oversee the

  3. Properties of vacuum brazed Si3N4/steel joint using active brazing filler metal

    NASA Astrophysics Data System (ADS)

    Bao, Fanghan; Ren, Jialie; Zhou, Yunhong; Yan, Ping

    The influence of active element Ti in Ag-Cu-Ti filler metal on wettability and joint strength is studied. Filler metal with 3 percent Ti achieves good results in direct brazing of silicon nitride to steel. For improving joint strength different interlayers are synthetically investigated. Experiments showed that an interlayer of low yield strength material can reduce the residual stress in the joint and increase joint strength more effectively than that of an interlayer of low coefficient of expansion material. Active element Ti in the brazing alloy diffused into Si3N4, and chemical reactions occurred in ceramic-metal interface producing certain chemical compounds consist of N and Ti.

  4. Crystal structure of inactive form of Rab3B

    SciTech Connect

    Zhang, Wei; Shen, Yang; Jiao, Ronghong; Liu, Yanli; Deng, Lingfu; Qi, Chao

    2012-06-28

    Rab proteins are the largest family of ras-related GTPases in eukaryotic cells. They act as directional molecular switches at membrane trafficking, including vesicle budding, cargo sorting, transport, tethering, and fusion. Here, we generated and crystallized the Rab3B:GDP complex. The structure of the complex was solved to 1.9 {angstrom} resolution and the structural base comparison with other Rab3 members provides a structural basis for the GDP/GTP switch in controlling the activity of small GTPase. The comparison of charge distribution among the members of Rab3 also indicates their different roles in vesicular trafficking.

  5. Specific interactions of Mss4 with members of the Rab GTPase subfamily.

    PubMed

    Burton, J L; Burns, M E; Gatti, E; Augustine, G J; De Camilli, P

    1994-12-01

    Mss4 is a mammalian protein that was identified as a suppressor of a yeast secretory mutant harboring a mutation in the GTPase Sec4 and was found to stimulate GDP release from this protein. We have now performed a biochemical characterization of the Mss4 protein and examined the specificity of its association with mammalian GTPases. Mss4 is primarily a soluble protein with a widespread tissue distribution. Recombinant Mss4 binds GTPases present in tissue extracts, and by a gel overlay assay binds specifically Rab Rab10proteins. We further define the Mss4-GTPase interaction to a subset of Rabs belonging to the same subfamily branch which include Rab1, Rab3, Rab8, Rab10, Sec4 and Ypt1 but not Rab2, Rab4, Rab5, Rab6, Rab9 and Rab11. Accordingly, Mss4 co-precipitates from a brain extract with Rab3a but not Rab5. Mss4 only stimulates GDP release from, and the association of GTP gamma S with, this Rab subset. Recombinant Mss4 and Rab3a form a stable complex in solution that is dissociated with either GDP or GTP gamma S. Injection of Mss4 into the squid giant nerve terminal enhances neurotransmitter release. These results suggest that Mss4 behaves as a guanylnucleotide exchange factor (GEF) for a subset of Rabs to influence distinct vesicular transport steps along the secretory pathway.

  6. Microstructural characterization of titanium to 304 stainless steel brazed joints

    SciTech Connect

    Camargo, P.R.C.; Liu, S. . Center for Welding and Joining Research); Trevisan, R.E. . Dept. of Fabrication Engineering)

    1993-12-01

    The formation of intermetallic compounds in brazed joints between titanium and 304 stainless steel is of major concern, since they considerably degrade the joint properties. This research examined the vacuum brazing of commercially pure titanium to 304 stainless steel using two different silver-copper brazing filler metals. Pure copper and silver were used to prepare the brazing filler metals in these experiments. Special attention was given to the characterization of the different phases formed at the brazed joint and the effect of the intermetallic compounds on the mechanical properties of the brazed joints. Light and electron microscopy, electron probe microanalysis, microhardness, and shear testing were used to support the investigation. From the mechanical properties point of view, brazed joints using an eutectic composition filler metal (Ag-28 wt-% Cu) proved to be superior compared to the joints prepared with a filler metal of composition Ag-46 wt-% Cu. To maximize the shear strength of the joint, the brazing time must be optimized such that interfacial reactions, titanium-iron intermetallics formation are minimized.

  7. Silver-palladium braze alloy recovered from masking materials

    NASA Technical Reports Server (NTRS)

    Cierniak, R.; Colman, G.; De Carlo, F.

    1966-01-01

    Method for recovering powdered silver-palladium braze alloy from an acrylic spray binder and rubber masking adhesive used in spray brazing is devised. The process involves agitation and dissolution of masking materials and recovery of suspended precious metal particles on a filter.

  8. Evaluation of Margins of Safety in Brazed Joints

    NASA Technical Reports Server (NTRS)

    Flom, Yury; Wang, Len; Powell, Mollie M.; Soffa, Matthew A.; Rommel, Monica L.

    2009-01-01

    One of the essential steps in assuring reliable performance of high cost critical brazed structures is the assessment of the Margin of Safety (MS) of the brazed joints. In many cases the experimental determination of the failure loads by destructive testing of the brazed assembly is not practical and cost prohibitive. In such cases the evaluation of the MS is performed analytically by comparing the maximum design loads with the allowable ones and incorporating various safety or knock down factors imposed by the customer. Unfortunately, an industry standard methodology for the design and analysis of brazed joints has not been developed. This paper provides an example of an approach that was used to analyze an AlBeMet 162 (38%Be-62%Al) structure brazed with the AWS BAlSi-4 (Al-12%Si) filler metal. A practical and conservative interaction equation combining shear and tensile allowables was developed and validated to evaluate an acceptable (safe) combination of tensile and shear stresses acting in the brazed joint. These allowables are obtained from testing of standard tensile and lap shear brazed specimens. The proposed equation enables the assessment of the load carrying capability of complex brazed joints subjected to multi-axial loading.

  9. Evaluation of methods for nondestructive testing of brazed joints

    NASA Technical Reports Server (NTRS)

    Kanno, A.

    1968-01-01

    Evaluation of nondestructive methods of testing brazed joints reveals that ultrasonic testing is effective in the detection of nonbonds in diffusion bonded samples. Radiography provides excellent resolutions of void or inclusion defects, and the neutron radiographic technique shows particular advantage for brazing materials containing cadmium.

  10. Flaw Tolerance in Lap Shear Brazed Joints. Part 1

    NASA Technical Reports Server (NTRS)

    Flom, Yury; Wang, Liqin; Obenschain, Arthur F. (Technical Monitor)

    2002-01-01

    Vacuum brazing is a viable process to achieve strong permanent and hermetic joints in space. As in any joining process, brazed Joints have various imperfections and defects. It is important to understand the impact that flaws have on the load carrying capacity and performance of the brazed joints. This study focuses on the behavior of lap shear joints due to their engineering importance in brazed aerospace structures. In Part 1 an average shear strength capabilitY and failure modes of the single lap joints are explored. Specimens comprised of 0.090 inch thick 347 stainless steel sheet brazed with pure silver are tested in accordance with the AWS C3.2 standard. Comparison of the measured loads and average shear stresses at failure with the Finite Element Analysis (FEA) of the same specimens as a function of the overlap lengths shows excellent correlation between the experimental and calculated values for the defect-free lap joints. In Part 2, various flaws will be introduced in the test specimens to simulate lack of braze coverage in the lap joints. Mechanical testing and FEA will be performed on these joints to verify if behavior of ductile ]appoints containing flaws is similar to the joints with the reduced braze area. Finally, in Part 3, the results obtained in Part 1 and 2 will be applied to the brazed assembly to evaluate a load carrying capability of the structural lap joint containing defects.

  11. Reactive Boride Brazing on Low-Alloy Automotive Grade Steel

    NASA Astrophysics Data System (ADS)

    Palanisamy, B.; Upadhyaya, A.

    2011-11-01

    Brazing is a widely used process to improve the performance of steels used in automotive applications. The substrate material is often exposed to harsh conditions in these applications and may affect the service life of the component. Reactive boride brazing aims to improve the mechanical properties of the substrate material by forming a ceramic-metal composite coating in a single-step process in situ. In this study, sintered Ancor 4300 low-alloy steel is used as the substrate with chromium-rich braze and chromium-lean braze materials. The mechanical properties of the brazed samples were studied in detail using microindentation hardness measurements and the transverse rupture test. The results indicate that the brazed superlayer has a 10 times higher hardness. There was a significant improvement in the transverse rupture strength of the steel brazed with the chromium-rich boride as compared to the pure substrate material. In an effort to reduce processing time, green compacts of the substrate were also directly brazed and yielded favorable results.

  12. Failure Assessment Diagram for Brazed 304 Stainless Steel Joints

    NASA Technical Reports Server (NTRS)

    Flom, Yory

    2011-01-01

    Interaction equations were proposed earlier to predict failure in Albemet 162 brazed joints. Present study demonstrates that the same interaction equations can be used for lower bound estimate of the failure criterion in 304 stainless steel joints brazed with silver-based filler metals as well as for construction of the Failure Assessment Diagrams (FAD).

  13. Tube dimpling tool assures accurate dip-brazed joints

    NASA Technical Reports Server (NTRS)

    Beuyukian, C. S.; Heisman, R. M.

    1968-01-01

    Portable, hand-held dimpling tool assures accurate brazed joints between tubes of different diameters. Prior to brazing, the tool performs precise dimpling and nipple forming and also provides control and accurate measuring of the height of nipples and depth of dimples so formed.

  14. Brazing method produces solid-solution bond between refractory metals

    NASA Technical Reports Server (NTRS)

    1965-01-01

    Brazing two refractory metals by diffusion bonding minimizes distortion and avoids excessive grain growth in the metals. This method requires the selection of an interface metal that forms intermediate low-melting eutectics or solid solutions with the metals to be brazed.

  15. Thermal response of ceramic components during electron beam brazing

    SciTech Connect

    Voth, T.E.; Gianoulakis, S.E.; Halbleib, J.A.

    1996-03-01

    Ceramics are being used increasingly in applications where high temperatures are encountered such as automobile and gas turbine engines. However, the use of ceramics is limited by a lack of methods capable of producing strong, high temperature joints. This is because most ceramic-ceramic joining techniques, such as brazing, require that the entire assembly be exposed to high temperatures in order to assure that the braze material melts. Alternatively, localized heating using high energy electron beams may be used to selectively heat the braze material. In this work, high energy electron beam brazing of a ceramic part is modeled numerically. The part considered consists of a ceramic cylinder and disk between which is sandwiched an annular washer of braze material. An electron beam impinges on the disk, melting the braze metal. The resulting coupled electron and thermal transport equations are solved using Monte Carlo and finite element techniques. Results indicate that increased electron beam current decreases time to melt as well as required cooling time. Vacuum furnace brazing was also simulated and predicted results indicate increased processing times relative to electron beam brazing.

  16. A Paired RNAi and RabGAP Overexpression Screen Identifies Rab11 as a Regulator of β-Amyloid Production

    PubMed Central

    Udayar, Vinod; Buggia-Prévot, Virginie; Guerreiro, Rita L.; Siegel, Gabriele; Rambabu, Naresh; Soohoo, Amanda L.; Ponnusamy, Moorthi; Siegenthaler, Barbara; Bali, Jitin; Guerreiro, Rita; Brás, José; Sassi, Celeste; Gibbs, J. Raphael; Hernandez, Dena; Lupton, Michelle K.; Brown, Kristelle; Morgan, Kevin; Powell, John; Singleton, Andrew; Hardy, John; Simons, Mikael; Ries, Jonas; Puthenveedu, Manojkumar A.; Hardy, John; Thinakaran, Gopal; Rajendran, Lawrence

    2014-01-01

    Summary Alzheimer’s disease (AD) is characterized by cerebral deposition of β-amyloid (Aβ) peptides, which are generated from amyloid precursor protein (APP) by β- and γ-secretases. APP and the secretases are membrane associated, but whether membrane trafficking controls Aβ levels is unclear. Here, we performed an RNAi screen of all human Rab-GTPases, which regulate membrane trafficking, complemented with a Rab-GTPase-activating protein screen, and present a road map of the membrane-trafficking events regulating Aβ production. We identify Rab11 and Rab3 as key players. Although retromers and retromer-associated proteins control APP recycling, we show that Rab11 controlled β-secretase endosomal recycling to the plasma membrane and thus affected Aβ production. Exome sequencing revealed a significant genetic association of Rab11A with late-onset AD, and network analysis identified Rab11A and Rab11B as components of the late-onset AD risk network, suggesting a causal link between Rab11 and AD. Our results reveal trafficking pathways that regulate Aβ levels and show how systems biology approaches can unravel the molecular complexity underlying AD. PMID:24373285

  17. Recombinant Leishmania Rab6 (rLdRab6) is recognized by sera from visceral leishmaniasis patients.

    PubMed

    Chauhan, Indira Singh; Shukla, Rantidev; Krishna, Shagun; Sekhri, Savita; Kaushik, Umesh; Baby, Sabitha; Pal, Chiranjib; Siddiqi, Mohammad Imran; Sundar, Shyam; Singh, Neeloo

    2016-11-01

    Rab proteins form the largest branch of the Ras superfamily. Rab proteins are key regulators of intracellular vesicular transport and membrane trafficking. Although RabGTPases are well-recognized targets in human diseases but are under-explored therapeutically in the Leishmania parasite. Using a quantitative cytofluorimetric assay, we analyzed the composition and organization of Rab6GTPase protein which was found to be primarily localized on the parasite subpellicular membrane and flagellum due to its association with kinesin motor proteins in the cytoskeletal microtubules. Our aim was to also assess the diagnostic role of recombinant Rab6 protein from Leishmania donovani (rLdRab6) using sera/plasma of Indian visceral leishmaniasis (VL) patients. Receiver-operating characteristic (ROC) curve analysis indicated 100% sensitivity and 100% specificity for rLdRab6-based ELISA which was almost similar in comparison to recombinant K39-based ELISA (95.83% sensitivity and 100% specificity). Sera of patients from another intracellular pathogenic infection, Mycobacterium tuberculosis, did not contain any significant levels of anti-rLdRab6 antibody. Thus rLdRab6 accuracy in visceral leishmaniasis diagnosis makes it a promising antigen for clinical use.

  18. Brazing of the Tore Supra actively cooled Phase III Limiter

    SciTech Connect

    Nygren, R.E.; Walker, C.A.; Lutz, T.J.; Hosking, F.M.; McGrath, R.T.

    1993-12-31

    The head of the water-cooled Tore Supra Phase 3 Limiter is a bank of 14 round OFHC copper tubes, curved to fit the plasma radius, onto which several hundred pyrolytic graphite (PG) tiles and a lesser number of carbon fiber composite tiles are brazed. The small allowable tolerances for fitting the tiles to the tubes and mating of compound curvatures made the brazing and fabrication extremely challenging. The paper describes the fabrication process with emphasis on the procedure for brazing. In the fixturing for vacuum furnace brazing, the tiles were each independently clamped to the tube with an elaborate set of window frame clamps. Braze quality was evaluated with transient heating tests. Some rebrazing was necessary.

  19. Thin-film diffusion brazing of titanium alloys

    NASA Technical Reports Server (NTRS)

    Mikus, E. B.

    1972-01-01

    A thin film diffusion brazing technique for joining titanium alloys by use of a Cu intermediate is described. The method has been characterized in terms of static and dynamic mechanical properties on Ti-6Al-4V alloy. These include tensile, fracture toughness, stress corrosion, shear, corrosion fatigue, mechanical fatigue and acoustic fatigue. Most of the properties of titanium joints formed by thin film diffusion brazing are equal or exceed base metal properties. The advantages of thin film diffusion brazing over solid state diffusion bonding and brazing with conventional braze alloys are discussed. The producibility advantages of this process over others provide the potential for producing high efficiency joints in structural components of titanium alloys for the minimum cost.

  20. Mechanical characterization and modeling of brazed EUROFER-tungsten-joints

    NASA Astrophysics Data System (ADS)

    Chehtov, T.; Aktaa, J.; Kraft, O.

    2007-08-01

    Within the scope of the European fusion power plant study for development of a He-cooled divertor, a tungsten-steel joint has been considered. A preferable joining technique is high temperature brazing. Brazed joints of dissimilar materials suffer from a mismatch in coefficients of thermal expansion. The components of the joint are exposed to mechanical and cyclic thermal loads which give rise to development of high stresses and could lead to failure. Brazed joints of tungsten alloy and ferritic-martensitic steel using different brazing filler materials were studied both experimentally and theoretically. Finite element computations have been performed to calculate the stress distribution and to investigate their evolution within the course of the operational thermal load. Sample joint specimen have been brazed, investigated with respect to their microstructure, and mechanically characterized by performing bend and notched bar impact testing at different temperatures. Some plastic deformation and relatively low impact energies were measured.

  1. Composition and method for brazing graphite to graphite

    DOEpatents

    Taylor, A.J.; Dykes, N.L.

    1982-08-10

    A brazing material is described for joining graphite structures that can be used up to 2800/sup 0/C. The brazing material is formed of a paste-like composition of hafnium carbide and uranium oxide with a thermosetting resin. The uranium oxide is converted to uranium dicarbide during the brazing operation and then the hafnium carbide and uranium dicarbide form a liquid phase at a temperature about 2600/sup 0/C with the uranium diffusing and vaporizing from the joint area as the temperature is increased to about 2800/sup 0/C so as to provide a brazed joint consisting essentially of hafnium carbide. The resulting brazed joint is chemically and thermally compatible with the graphite structures.

  2. Flaw Tolerance in Lap Shear Brazed Joints. Part 1

    NASA Technical Reports Server (NTRS)

    Flom, Yury; Wang, Li-Qin

    2003-01-01

    Furnace brazing is a joining process used in the aerospace and other industries to produce strong permanent and hermetic structural joints. As in any joining process, brazed joints have various imperfections and defects. At the present time, our understanding of the influence of the internal defects on the strength of the brazed joints is not adequate. The goal of this 3-part investigation is to better understand the properties and failure mechanisms of the brazed joints containing defects. This study focuses on the behavior of the brazed lap shear joints because of their importance in manufacturing aerospace structures. In Part 1, an average shear strength capability and failure modes of the single lap joints are explored. Stainless steel specimens brazed with pure silver are tested in accordance with the AWS C3.2 standard. Comparison of the failure loads and the ultimate shear strength with the Finite Element Analysis (FEA) of the same specimens as a function of the overlap widths shows excellent correlation between the experimental and calculated values for the defect-free lap joints. A damage zone criterion is shown to work quite well in understanding the failure of the braze joints. In Part 2, the findings of the Part 1 will be verified on the larger test specimens. Also, various flaws will be introduced in the test specimens to simulate lack of braze coverage in the lap joints. Mechanical testing and FEA will be performed on these joints to verify that behavior of the flawed ductile lap joints is similar to joints with a reduced braze area. Finally, in Part 3, the results obtained in Parts 1 and 2 will be applied to the actual brazed structure to evaluate the load-carrying capability of a structural lap joint containing discontinuities. In addition, a simplified engineering procedure will be offered for the laboratory testing of the lap shear specimens.

  3. Small GTPase Rab2B and Its Specific Binding Protein Golgi-associated Rab2B Interactor-like 4 (GARI-L4) Regulate Golgi Morphology*

    PubMed Central

    Aizawa, Megumi; Fukuda, Mitsunori

    2015-01-01

    Rab small GTPases are crucial regulators of the membrane traffic that maintains organelle identity and morphology. Several Rab isoforms are present in the Golgi, and it has been suggested that they regulate the compacted morphology of the Golgi in mammalian cells. However, the functional relationships among the Golgi-resident Rabs, e.g. whether they are functionally redundant or different, are poorly understood. In this study, we used specific siRNAs to perform genome-wide screening for human Rabs that are involved in Golgi morphology in HeLa-S3 cells. The results showed that knockdown of any one of the six Rab isoforms (Rab1A/1B/2A/2B/6B/8A) induced fragmentation of the Golgi in HeLa-S3 cells and that its phenotype was rescued by re-expression of their respective siRNA-resistant construct. We then performed systematic knockdown-rescue experiments in relation to each of the six Rabs. Interestingly, with the exception of the Rab8A knockdown, the Golgi fragmentation phenotype induced by knockdown of a single Rab isoform, e.g. Rab2B, was efficiently rescued by re-expression of its siRNA-resistant Rab alone, not by any of the other five Rabs, e.g. Rab2A, which is highly homologous to Rab2B, indicating that these Rab isoforms non-redundantly regulate Golgi morphology possibly through interaction with isoform-specific effector molecules. In addition, we identified Golgi-associated Rab2B interactor-like 4 (GARI-L4) as a novel Golgi-resident Rab2B-specific binding protein whose knockdown also induced fragmentation of the Golgi. Our findings suggest that the compacted Golgi morphology of mammalian cells is finely tuned by multiple sets of Rab (or Rab-effector complexes) that for the most part function independently. PMID:26209634

  4. Large Area Active Brazing of Multi-tile Ceramic-Metal Structures

    DTIC Science & Technology

    2012-05-01

    metal brazing . The active brazing alloys wet most materials (including ceramics and corrosion- resistant metals such as titanium alloys and stainless...bonding in ceramic-metal systems is active metal brazing . The active brazing alloys wet most materials (including ceramics and corrosion-resistant...an idea from the ‘50s by putting an “active” component, such as titanium, directly into a brazing alloy , typically a silver-copper eutectic, to

  5. RAB24 facilitates clearance of autophagic compartments during basal conditions

    PubMed Central

    Ylä-Anttila, Päivi; Mikkonen, Elisa; Happonen, Kaisa E; Holland, Petter; Ueno, Takashi; Simonsen, Anne; Eskelinen, Eeva-Liisa

    2015-01-01

    RAB24 belongs to a family of small GTPases and has been implicated to function in autophagy. Here we confirm the intracellular localization of RAB24 to autophagic vacuoles with immuno electron microscopy and cell fractionation, and show that prenylation and guanine nucleotide binding are necessary for the targeting of RAB24 to autophagic compartments. Further, we show that RAB24 plays a role in the maturation and/or clearance of autophagic compartments under nutrient-rich conditions, but not during short amino acid starvation. Quantitative electron microscopy shows an increase in the numbers of late autophagic compartments in cells silenced for RAB24, and mRFP-GFP-LC3 probe and autophagy flux experiments indicate that this is due to a hindrance in their clearance. Formation of autophagosomes is shown to be unaffected by RAB24-silencing with siRNA. A defect in aggregate clearance in the absence of RAB24 is also shown in cells forming polyglutamine aggregates. This study places RAB24 function in the termination of the autophagic process under nutrient-rich conditions. PMID:26325487

  6. Rab proteins: The key regulators of intracellular vesicle transport

    SciTech Connect

    Bhuin, Tanmay; Roy, Jagat Kumar

    2014-10-15

    Vesicular/membrane trafficking essentially regulates the compartmentalization and abundance of proteins within the cells and contributes in many signalling pathways. This membrane transport in eukaryotic cells is a complex process regulated by a large and diverse array of proteins. A large group of monomeric small GTPases; the Rabs are essential components of this membrane trafficking route. Most of the Rabs are ubiquitously expressed proteins and have been implicated in vesicle formation, vesicle motility/delivery along cytoskeleton elements and docking/fusion at target membranes through the recruitment of effectors. Functional impairments of Rabs affecting transport pathways manifest different diseases. Rab functions are accompanied by cyclical activation and inactivation of GTP-bound and GDP-bound forms between the cytosol and membranes which is regulated by upstream regulators. Rab proteins are characterized by their distinct sub-cellular localization and regulate a wide variety of endocytic, transcytic and exocytic transport pathways. Mutations of Rabs affect cell growth, motility and other biological processes. - Highlights: • Rab proteins regulate different signalling pathways. • Deregulation of Rabs is the fundamental causes of a variety of human diseases. • This paper gives potential directions in developing therapeutic targets. • This paper also gives ample directions for modulating pathways central to normal physiology. • These are the huge challenges for drug discovery and delivery in near future.

  7. Rubicon controls endosome maturation as a Rab7 effector.

    PubMed

    Sun, Qiming; Westphal, Wiebke; Wong, Kwun Ngok; Tan, Irena; Zhong, Qing

    2010-11-09

    The activation and recruitment of the small GTPase Rab7 to early endosome is a critical step for early to late endosome maturation, a process that requires the class III phosphatidylinositol 3-kinase (PI3KC3) and GTPase regulators. However, the molecular mechanism underlying Rab7 activation and endosome maturation is still poorly defined. Here we report that Rubicon, a component of the PI3KC3 complex, prevents endosome maturation through differential interactions with Rab7 and UVRAG. UVRAG activates PI3KC3 and C-VPS/HOPS, a guanine nucleotide exchange factor that catalyzes the exchange of GDP for GTP on Rab7. We demonstrate that Rubicon sequesters UVRAG from C-VPS/HOPS. Active GTP-bound Rab7 competes for Rubicon binding and releases UVRAG to associate with C-VPS/HOPS, which in turn promotes further loading of Rab7 with GTP. This feed-forward loop ensures rapid amplification of GTP-bound Rab7 and consequent stimulation of endosome maturation. Hence, Rubicon serves as a previously unknown Rab7 effector to ensure the proper progression of the endocytic pathway.

  8. Multivalent Rab interactions determine tether-mediated membrane fusion

    PubMed Central

    Lürick, Anna; Gao, Jieqiong; Kuhlee, Anne; Yavavli, Erdal; Langemeyer, Lars; Perz, Angela; Raunser, Stefan; Ungermann, Christian

    2017-01-01

    Membrane fusion at endomembranes requires cross-talk between Rab GTPases and tethers to drive SNARE-mediated lipid bilayer mixing. Several tethers have multiple Rab-binding sites with largely untested function. Here we dissected the lysosomal HOPS complex as a tethering complex with just two binding sites for the Rab7-like Ypt7 protein to determine their relevance for fusion. Using tethering and fusion assays combined with HOPS mutants, we show that HOPS-dependent fusion requires both Rab-binding sites, with Vps39 being the stronger Ypt7 interactor than Vps41. The intrinsic amphipathic lipid packaging sensor (ALPS) motif within HOPS Vps41, a target of the vacuolar kinase Yck3, is dispensable for tethering and fusion but can affect tethering if phosphorylated. In combination, our data demonstrate that a multivalent tethering complex uses its two Rab bindings to determine the place of SNARE assembly and thus fusion at endomembranes. PMID:27852901

  9. Macroblock manufacturing by 'hip assisted brazing' method

    NASA Astrophysics Data System (ADS)

    Chaumat, G.; Le Gallo, P.; Le Marois, G.; Moret, F.; Deschamp, P.

    1996-10-01

    The aim of this work was to demonstrate the potential of an original technology which allows the manufacturing of large size so-called carbon fibre composite 'macroblock' tube-in-tile components associated with a reliability for a future industrial production. This process includes the combinations of two techniques: (a) a hot isostatic pressing cycle to allow intimate contact of a copper tube with a CFC block during all the process; (b) a brazing operation during the HIP cycle to perform a good joint between both substrates. Finally, two prototypes have been realized and a non destructive evaluation of the quality of the joint has been carried out.

  10. Rab27a Targeting to Melanosomes Requires Nucleotide Exchange but Not Effector Binding

    PubMed Central

    Tarafder, Abul K; Wasmeier, Christina; Figueiredo, Ana C; Booth, Antonia E G; Orihara, Asumi; Ramalho, Jose S; Hume, Alistair N; Seabra, Miguel C

    2011-01-01

    Rab GTPases are important determinants of organelle identity and regulators of vesicular transport pathways. Consequently, each Rab occupies a highly specific subcellular localization. However, the precise mechanisms governing Rab targeting remain unclear. Guanine nucleotide exchange factors (GEFs), putative membrane-resident targeting factors and effector binding have all been implicated as critical regulators of Rab targeting. Here, we address these issues using Rab27a targeting to melanosomes as a model system. Rab27a regulates motility of lysosome-related organelles and secretory granules. Its effectors have been characterized extensively, and we have identified Rab3GEP as the non-redundant Rab27a GEF in melanocytes (Figueiredo AC et al. Rab3GEP is the non-redundant guanine nucleotide exchange factor for Rab27a in melanocytes. J Biol Chem 2008;283:23209–23216). Using Rab27a mutants that show impaired binding to representatives of all four Rab27a effector subgroups, we present evidence that effector binding is not essential for targeting of Rab27a to melanosomes. In contrast, we observed that knockdown of Rab3GEP resulted in mis-targeting of Rab27a, suggesting that Rab3GEP activity is required for correct targeting of Rab27a. However, the identification of Rab27a mutants that undergo efficient GDP/GTP exchange in the presence of Rab3GEP in vitro but are mis-targeted in a cellular context indicates that nucleotide loading is not the sole determinant of subcellular targeting of Rab27a. Our data support a model in which exchange activity, but not effector binding, represents one essential factor that contributes to membrane targeting of Rab proteins. PMID:21554507

  11. Coating system to permit direct brazing of ceramics

    DOEpatents

    Cadden, Charles H.; Hosking, F. Michael

    2003-01-01

    This invention relates to a method for preparing the surface of a ceramic component that enables direct brazing using a non-active braze alloy. The present invention also relates to a method for directly brazing a ceramic component to a ceramic or metal member using this method of surface preparation, and to articles produced by using this brazing method. The ceramic can be high purity alumina. The method comprises applying a first coating of a silicon-bearing oxide material (e.g. silicon dioxide or mullite (3Al.sub.2 O.sub.3.2SiO.sub.2) to the ceramic. Next, a thin coating of active metal (e.g. Ti or V) is applied. Finally, a thicker coating of a non-active metal (e.g. Au or Cu) is applied. The coatings can be applied by physical vapor deposition (PVD). Alternatively, the active and non-active metals can be co-deposited (e.g. by sputtering a target made of mullite). After all of the coatings have been applied, the ceramic can be fired at a high temperature in a non-oxidizing environment to promote diffusion, and to enhance bonding of the coatings to the substrate. After firing, the metallized ceramic component can be brazed to other components using a conventional non-active braze alloy. Alternatively, the firing and brazing steps can be combined into a single step. This process can replace the need to perform a "moly-manganese" metallization step.

  12. Effect of filler metal composition on the strength of yttria stabilized zirconia joints brazed with Pd-Ag-CuOx

    SciTech Connect

    Darsell, Jens T.; Weil, K. Scott

    2008-09-08

    The Ag-CuOx system is of interest to be used to be used as an air braze filler metal for joining high temperature electrochemical devices. Previous work has shown that the melting temperatures can be increased by adding palladium to Ag-CuOx and it is expected that this may aid high temperature stability. This work compares the room temperature bend strength of joints made between yttria-stabilized zirconia (YSZ) air brazed using Ag-CuOx without palladium and with 5 and 15mol% palladium additions. It has been found that in general palladium decreases joint strength, especially in low copper oxide compositions filler metals. At high copper oxide contents, brittle fracture through both copper oxide rich phases and the YSZ limits joint strength.

  13. Rab5 Isoforms Specifically Regulate Different Modes of Endocytosis in Leishmania.

    PubMed

    Rastogi, Ruchir; Verma, Jitender Kumar; Kapoor, Anjali; Langsley, Gordon; Mukhopadhyay, Amitabha

    2016-07-08

    Differential functions of Rab5 isoforms in endocytosis are not well characterized. Here, we cloned, expressed, and characterized Rab5a and Rab5b from Leishmania and found that both of them are localized in the early endosome. To understand the role of LdRab5 isoforms in different modes of endocytosis in Leishmania, we generated transgenic parasites overexpressing LdRab5a, LdRab5b, or their dominant-positive (LdRab5a:Q93L and LdRab5b:Q80L) or dominant-negative mutants (LdRab5a:N146I and LdRab5b:N133I). Using LdRab5a or its mutants overexpressing parasites, we found that LdRab5a specifically regulates the fluid-phase endocytosis of horseradish peroxidase and also specifically induced the transport of dextran-Texas Red to the lysosomes. In contrast, cells overexpressing LdRab5b or its mutants showed that LdRab5b explicitly controls receptor-mediated endocytosis of hemoglobin, and overexpression of LdRab5b:WT enhanced the transport of internalized Hb to the lysosomes in comparison with control cells. To unequivocally demonstrate the role of Rab5 isoforms in endocytosis in Leishmania, we tried to generate null-mutants of LdRab5a and LdRab5b parasites, but both were lethal indicating their essential functions in parasites. Therefore, we used heterozygous LdRab5a(+/-) and LdRab5b(+/-) cells. LdRab5a(+/-) Leishmania showed 50% inhibition of HRP uptake, but hemoglobin endocytosis was uninterrupted. In contrast, about 50% inhibition of Hb endocytosis was observed in LdRab5b(+/-) cells without any significant effect on HRP uptake. Finally, we tried to identify putative LdRab5a and LdRab5b effectors. We found that LdRab5b interacts with clathrin heavy chain and hemoglobin receptor. However, LdRab5a failed to interact with the clathrin heavy chain, and interaction with hemoglobin receptor was significantly less. Thus, our results showed that LdRab5a and LdRab5b differentially regulate fluid phase and receptor-mediated endocytosis in Leishmania.

  14. Microstructure and Mechanical Properties of Joints in Sintered SiC Fiber-Bonded Ceramics Brazed with Ag-Cu-Ti Alloy

    SciTech Connect

    Singh, Mrityunjay; Matsunaga, Tadashi; Lin, Hua-Tay; Asthana, Rajiv; Ishikawa, Toshihiro

    2012-01-01

    Active metal brazing of a new high thermal conductivity sintered SiC-polycrystalline fiber-bonded ceramic (SA-Tyrannohex{reg_sign}) has been carried out using a Ti-containing Ag-Cu active braze alloy (Cusil-ABA{reg_sign}). The brazed composite joints were characterized using scanning electron microscopy coupled with energy-dispersive X-ray spectrometry (SEM-EDS). The results show that this material can be successfully joined using judiciously selected off-the shelf active braze alloys to yield metallurgically sound joints possessing high integrity. Uniform and continuous joints were obtained irrespective of differences in the fiber orientation in the substrate material. Detailed interfacial microanalysis showed that the titanium reacts with C and Si to form TiC layer and a Ti-Si compound, respectively. Furthermore, the evaluation of shear strength of the joints was also conducted at ambient and elevated temperatures in air using the single-lap offset (SLO) shear test. The perpendicular-type SA-Tyrannohex joints exhibited apparent shear strengths of about 42 MPa and 25 MPa at 650 C and 750 C, respectively. The fracture at the higher temperature occurred at the interface between the reaction-formed TiC layer and braze. This might be caused by generation of stress intensity when a shear stress was applied, according to {mu}-FEA simulation results.

  15. Microstructure and mechanical properties of joints in sintered SiC fiber-bonded ceramics brazed with Ag Cu Ti alloy

    SciTech Connect

    Singh, Mrityunjay; Asthana, Rajiv; Ishikawa, Toshihiro; Matsunaga, Tadashi; Lin, Hua-Tay

    2012-01-01

    Active metal brazing of a new high thermal conductivity sintered SiC-polycrystalline fiber-bonded ceramic (SA-Tyrannohexs) has been carried out using a Ti-containing Ag Cu active braze alloy (Cusil-ABAs). The brazed composite joints were characterized using scanning electron microscopy coupled with energy-dispersive X-ray spectrometry (SEM EDS). The results show that this material can be successfully joined using judiciously selected off-the shelf active braze alloys to yield metallurgically sound joints possessing high integrity. Uniform and continuous joints were obtained irrespective of differences in the fiber orientation in the substrate material. Detailed interfacial microanalysis showed that the titanium reacts with C and Si to form TiC layer and a Ti Si compound, respectively. Furthermore, the evaluation of shear strength of the joints was also conducted at ambient and elevated temperatures in air using the single-lap offset (SLO) shear test. The perpendicular-type SA-Tyrannohex joints exhibited apparent shear strengths of about 42 MPa and 25 MPa at 650 1C and 750 1C, respectively. The fracture at the higher temperature occurred at the interface between the reactionformed TiC layer and braze. This might be caused by generation of stress intensity when a shear stress was applied, according to m-FEA simulation results.

  16. Coaxial Control of Aluminum and Steel Laser Brazing Processes

    NASA Astrophysics Data System (ADS)

    Frank, Sascha; Ungers, Michael; Rolser, Raphael

    Laser brazing has become firmly established as a joining process in the automotive industry. While this process offers many advantages, brazed seams also have to meet particularly high quality requirements. The challenge of creating a suitable online quality control system is magnified by the increasing use of aluminum in automotive engineering. This paper introduces recent works on the development of a system for controlling both aluminum and steel brazing processes. It also discusses some of the challenges connected to this task and discloses some of the results derived from the corresponding process analysis.

  17. Plasma Spray for Difficult-To-Braze Alloys

    NASA Technical Reports Server (NTRS)

    Brennan, A.

    1982-01-01

    Nickel plating on surfaces makes brazing easier for some alloys. Sometimes nickel plating may not be feasible because of manufacturing sequence, size of hardware, or lack of suitable source for nickel plating. Alternative surface preparation in such cases is to grit-blast surface lightly and then plasma-spray 1 1/2 to 2 mils of fine nickel powder or braze-alloy material directly on surface. Powder is sprayed from plasma gun, using argon as carrier gas to prevent oxidation of nickel or braze alloy.

  18. Brazing copper to dispersion-strengthened copper

    SciTech Connect

    Ryding, D.G.; Allen, D.; Lee, R.

    1996-08-01

    The Advanced Photon Source (APS) is a state-of-the-art synchrotron light source that will produce intense x-ray beams, which will allow the study of smaller samples and faster reactions and processes at a greater level of detail that has been possible to date. The beam is produced by using third-generation insertion devices in a 7 GeV electron/positron storage ring that is 1100 meters in circumference. The heat load from these intense high power devices is very high and certain components must sustain total heat loads of 3 to 15 kW and heat fluxes of 30 W/mm{sup 2}. Because the beams will cycle on and off many times, thermal shock and fatigue will be a problem. High heat flux impinging on a small area causes a large thermal gradient that results in high stress. GlidCop{reg_sign}, a dispersion strengthened copper, is the desired material because of its high thermal conductivity and superior mechanical properties as compared to copper and its alloys. GlidCop is not amenable to joining by fusion welding, and brazing requires diligence because of high diffusivity. Brazing procedures were developed using optical and scanning electron microscopy.

  19. Rene 95 brazed joint metallurgical program

    NASA Technical Reports Server (NTRS)

    Gay, C.; Givens, J.; Mastrorroco, S.; Sterman, A.

    1972-01-01

    This metallurgical program was specifically conducted for the establishment of material properties required for the design of the LF460 fan. The LF460 lift fan is an advanced 18:1 high thrust to weight single stage design. It has a turbine attached to the outer flowpath of the fan blade tip which minimizes the axial depth of the fan. Advanced lightweight attachment designs are employed in this concept to achieve minimum mass polar moments of inertia which are required for good aircraft flight response control. The design features which are unique to this advanced LF460 lift fan are the 0.010 inch thin Udimet 700 alloy integral tip turbine design, minimum weight braze attachment of the turbine to the fan blade, and the high strength and elevated temperature capability of the Rene'95 alloy for the fan blade. The data presented in this report show that the LF460 fan rotor design is feasible and that the design stresses and margins of safety were more than adequate. Prior to any production application, however, additional stress rupture/shear lap joints should be run in order to establish a firm 1200 F stress rupture curve for the CM50 braze metal.

  20. Cathodic ARC surface cleaning prior to brazing

    SciTech Connect

    Dave, V. R.; Hollis, K. J.; Castro, R. G.; Smith, F. M.; Javernick, D. A.

    2002-01-01

    Surface cleanliness is one the critical process variables in vacuum furnace brazing operations. For a large number of metallic components, cleaning is usually accomplished either by water-based alkali cleaning, but may also involve acid etching or solvent cleaning / rinsing. Nickel plating may also be necessary to ensure proper wetting. All of these cleaning or plating technologies have associated waste disposal issues, and this article explores an alternative cleaning process that generates minimal waste. Cathodic arc, or reserve polarity, is well known for welding of materials with tenacious oxide layers such as aluminum alloys. In this work the reverse polarity effect is used to clean austenitic stainless steel substrates prior to brazing with Ag-28%Cu. This cleaning process is compared to acid pickling and is shown to produce similar wetting behavior as measured by dynamic contact angle experiments. Additionally, dynamic contact angle measurements with water drops are conducted to show that cathodic arc cleaning can remove organic contaminants as well. The process does have its limitations however, and alloys with high titanium and aluminum content such as nickel-based superalloys may still require plating to ensure adequate wetting.

  1. bMERB domains are bivalent Rab8 family effectors evolved by gene duplication.

    PubMed

    Rai, Amrita; Oprisko, Anastasia; Campos, Jeremy; Fu, Yangxue; Friese, Timon; Itzen, Aymelt; Goody, Roger S; Gazdag, Emerich Mihai; Müller, Matthias P

    2016-08-23

    In their active GTP-bound form, Rab proteins interact with proteins termed effector molecules. In this study, we have thoroughly characterized a Rab effector domain that is present in proteins of the Mical and EHBP families, both known to act in endosomal trafficking. Within our study, we show that these effectors display a preference for Rab8 family proteins (Rab8, 10, 13 and 15) and that some of the effector domains can bind two Rab proteins via separate binding sites. Structural analysis allowed us to explain the specificity towards Rab8 family members and the presence of two similar Rab binding sites that must have evolved via gene duplication. This study is the first to thoroughly characterize a Rab effector protein that contains two separate Rab binding sites within a single domain, allowing Micals and EHBPs to bind two Rabs simultaneously, thus suggesting previously unknown functions of these effector molecules in endosomal trafficking.

  2. Rab2a and Rab27a cooperatively regulate the transition from granule maturation to exocytosis through the dual effector Noc2.

    PubMed

    Matsunaga, Kohichi; Taoka, Masato; Isobe, Toshiaki; Izumi, Tetsuro

    2017-02-01

    Exocytosis of secretory granules entails budding from the trans-Golgi network, sorting and maturation of cargo proteins, and trafficking and fusion to the plasma membrane. Rab27a regulates the late steps in this process, such as granule recruitment to the fusion site, whereas Rab2a functions in the early steps, such as granule biogenesis and maturation. Here, we demonstrate that these two small GTPases simultaneously bind to Noc2 (also known as RPH3AL) in a GTP-dependent manner, although Rab2a binds only after Rab27a has bound. In pancreatic β-cells, the ternary Rab2a-Noc2-Rab27a complex specifically localizes on perinuclear immature granules, whereas the binary Noc2-Rab27a complex localizes on peripheral mature granules. In contrast to the wild type, Noc2 mutants defective in binding to Rab2a or Rab27a fail to promote glucose-stimulated insulin secretion. Although knockdown of any component of the ternary complex markedly inhibits insulin secretion, only knockdown of Rab2a or Noc2, and not that of Rab27a, impairs cargo processing from proinsulin to insulin. These results suggest that the dual effector, Noc2, regulates the transition from Rab2a-mediated granule biogenesis to Rab27a-mediated granule exocytosis.

  3. Braze system and method for reducing strain in a braze joint

    DOEpatents

    Cadden, Charles H.; Goods, Steven H.; Prantil, Vincent C.

    2004-05-11

    A system for joining a pair of structural members having widely differing coefficients of thermal expansion is disclosed. A mechanically "thick" foil is made by dispersing a refractory metal powder, such as molybdenum, niobium, tantalum, or tungsten into a quantity of a liquid, high expansion metal such as copper, silver, or gold, casting an ingot of the mixture, and then cutting sections of the ingot about 1 mm thick to provide the foil member. These foil members are shaped, and assembled between surfaces of structural members for joining, together with a layer of a braze alloy on either side of the foil member capable of wetting both the surfaces of the structural members and the foil. The assembled body is then heated to melt the braze alloy and join the assembled structure. The foil member subsequently absorbs the mechanical strain generated by the differential contraction of the cooling members that results from the difference in the coefficients of thermal expansion of the members.

  4. Soldering and brazing safety guide: A handbook on space practice for those involved in soldering and brazing

    NASA Astrophysics Data System (ADS)

    This manual provides those involved in welding and brazing with effective safety procedures for use in performance of their jobs. Hazards exist in four types of general soldering and brazing processes: (1) cleaning; (2) application of flux; (3) application of heat and filler metal; and (4) residue cleaning. Most hazards during those operations can be avoided by using care, proper ventilation, protective clothing and equipment. Specific process hazards for various methods of brazing and soldering are treated. Methods to check ventilation are presented as well as a check of personal hygiene and good maintenance practices are stressed. Several emergency first aid treatments are described.

  5. Rab20 regulates phagosome maturation in RAW264 macrophages during Fc gamma receptor-mediated phagocytosis.

    PubMed

    Egami, Youhei; Araki, Nobukazu

    2012-01-01

    Rab20, a member of the Rab GTPase family, is known to be involved in membrane trafficking, however its implication in FcγR-mediated phagocytosis is unclear. We examined the spatiotemporal localization of Rab20 during phagocytosis of IgG-opsonized erythrocytes (IgG-Es) in RAW264 macrophages. By the live-cell imaging of fluorescent protein-fused Rab20, it was shown that Rab20 was transiently associated with the phagosomal membranes. During the early stage of phagosome formation, Rab20 was not localized on the membranes of phagocytic cups, but was gradually recruited to the newly formed phagosomes. Although Rab20 was colocalized with Rab5 to some extent, the association of Rab20 with the phagosomes persisted even after the loss of Rab5 from the phagosomal membranes. Then, Rab20 was colocalized with Rab7 and Lamp1, late endosomal/lysosomal markers, on the internalized phagosomes. Moreover, our analysis of Rab20 mutant expression revealed that the maturation of phagosomes was significantly delayed in cells expressing the GDP-bound mutant Rab20-T19N. These data suggest that Rab20 is an important component of phagosome and regulates the phagosome maturation during FcγR-mediated phagocytosis.

  6. Low vapor pressure braze alloys for thermionic energy converters

    NASA Technical Reports Server (NTRS)

    Bair, V. L.

    1976-01-01

    The evaluation of cesium diode electrode materials called for braze fillers with very low vapor pressures and a wide range of melting points. Binary alloys of low vapor pressure refractory metals were chosen to fill this need. These alloys of Th, Zr, Hf, Ru, Nb, Ir, Mo, Ta, Os, Re, and W have reported melting point minima or eutectics from 1,510 K to above 3,000 K. Preliminary data are compiled on the use of several of these braze alloys. Melting points and surface wetting on a Ta base are given. Results of brazing Ir, LaB6, Nb, Re, W, and Zr-22 wt % ZrO2 materials into Ta and Nb-1% Zr bases are presented. Current braze usage is summarized.

  7. Failure Assessment of Stainless Steel and Titanium Brazed Joints

    NASA Technical Reports Server (NTRS)

    Flom, Yury A.

    2012-01-01

    Following successful application of Coulomb-Mohr and interaction equations for evaluation of safety margins in Albemet 162 brazed joints, two additional base metal/filler metal systems were investigated. Specimens consisting of stainless steel brazed with silver-base filler metal and titanium brazed with 1100 Al alloy were tested to failure under combined action of tensile, shear, bending and torsion loads. Finite Element Analysis (FEA), hand calculations and digital image comparison (DIC) techniques were used to estimate failure stresses and construct Failure Assessment Diagrams (FAD). This study confirms that interaction equation R(sub sigma) + R(sub tau) = 1, where R(sub sigma) and R(sub t u) are normal and shear stress ratios, can be used as conservative lower bound estimate of the failure criterion in stainless steel and titanium brazed joints.

  8. Control of vacuum induction brazing system for sealing of instrumentation feed-through

    SciTech Connect

    Sung Ho Ahn; Jintae Hong; Chang Young Joung; Ka Hae Kim; Sung Ho Heo

    2015-07-01

    The integrity of instrumentation cables is an important performance parameter in addition to the sealing performance in the brazing process. An accurate brazing control was developed for the brazing of the instrumentation feed-through in the vacuum induction brazing system in this paper. The experimental results show that the accurate brazing temperature control performance is achieved by the developed control scheme. Consequently, the sealing performances of the instrumentation feed-through and the integrities of the instrumentation cables were satisfied after brazing. (authors)

  9. Cryogenic performance of slotted brazed Rene 41 honeycomb panels

    NASA Technical Reports Server (NTRS)

    Hepler, A. K.; Swegle, A. R.

    1982-01-01

    Two brazed Rene 41 honeycomb panels that would incorporate a frame element were designed, fabricated and tested. The panels were representative of the lower surface of an advanced space transportation vehicle. The first panel was a two span panel supported by a center frame and on edges parallel to it. The second panel was a two span panel supported by a center frame and on edges parallel to it. The second panel was a three span panel supported on two frames and on edges parallel to the frames. Each panel had its outer skin slotted to reduce the thermal stresses of the panel skins. The first panel was tested under simulated boost conditions that included liquid hydrogen exposure of the frame and inner skin and radiant heat to 478K on the outer skins. The first panel was tested to investigate the effect of thermal stresses in skins and core caused by the panel being restrained by a cold integral frame and to observe the effects of frost formation and possible liquid air development in and around outer skin slots.

  10. Ceramic-to-metal stator vane assembly with braze

    DOEpatents

    Chase, Donna J.; Fang, Ho T.; Irwin, Craig W.; Schienle, James L.

    1995-01-01

    A stator vane assembly for a gas turbine engine that includes a plurality of circumferentially spaced ceramic vanes, each of which has an inner and outer ceramic shroud, and a ceramic post extending from one of the shrouds, and a metallic platform having a plurality of circumferentially spaced recesses. The posts are inserted into a metallic sleeve and then brazed. The brazed sleeves are then mounted in the recesses. A method for assembling these components to form the stator assembly is also described.

  11. Reactive Brazing of Carbon-Carbon Composites to Titanium

    NASA Technical Reports Server (NTRS)

    Shpargel, Tarah; Singh, M.; Morscher, Gregory; Asthana, Rajiv

    2004-01-01

    The Ti-metal/C-C composite joints were formed by reactive brazing with three commercial brazes, namely, Cu-ABA, TiCuNi, and TiCuSil. The joint microstructures were examined using optical microscopy, and scanning electron microscopy (SEM) coupled with energy dispersive spectrometry (EDS). The results of the microstructure analysis indicate solute redistribution across the joint and possible metallurgical bond formation via interdiffusion, which led to good wetting and spreading.

  12. Improved Assembly for Gas Shielding During Welding or Brazing

    NASA Technical Reports Server (NTRS)

    Gradl, Paul; Baker, Kevin; Weeks, Jack

    2009-01-01

    An improved assembly for inert-gas shielding of a metallic joint is designed to be useable during any of a variety of both laser-based and traditional welding and brazing processes. The basic purpose of this assembly or of a typical prior related assembly is to channel the flow of a chemically inert gas to a joint to prevent environmental contamination of the joint during the welding or brazing process and, if required, to accelerate cooling upon completion of the process.

  13. Active Metal Brazing of Carbon-Carbon Composites to Titanium

    NASA Technical Reports Server (NTRS)

    Singh, M.; Shpargel, T. P.; Morscher, G.; Asthana, R.

    2004-01-01

    The Ti-metal/C-C composite joints were formed by reactive brazing with three commercial brazes, namely, Cu-ABA, TiCuNi, and TiCuSil. The joint microstructures were examined using optical microscopy, and scanning electron microscopy (SEM) coupled with energy dispersive spectrometry (EDS). The results of the microstructure analysis indicate solute redistribution across the joint which led to good wetting, spreading, and metallurgical bond formation via interdiffusion.

  14. Functional Characterization of Monomeric GTPase Rab1 in the Secretory Pathway of Leishmania*

    PubMed Central

    Bahl, Surbhi; Parashar, Smriti; Malhotra, Himanshu; Raje, Manoj; Mukhopadhyay, Amitabha

    2015-01-01

    Leishmania secretes a large number of its effectors to the extracellular milieu. However, regulation of the secretory pathway in Leishmania is not well characterized. Here, we report the cloning, expression, and characterization of the Rab1 homologue from Leishmania. We have found that LdRab1 localizes in Golgi in Leishmania. To understand the role of LdRab1 in the secretory pathway of Leishmania, we have generated transgenic parasites overexpressing GFP-LdRab1:WT, GFP-LdRab1:Q67L (a GTPase-deficient dominant positive mutant of Rab1), and GFP-LdRab1:S22N (a GDP-locked dominant negative mutant of Rab1). Surprisingly, our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N does not disrupt the trafficking and localization of hemoglobin receptor in Leishmania. To determine whether the Rab1-dependent secretory pathway is conserved in parasites, we have analyzed the role of LdRab1 in the secretion of secretory acid phosphatase and Ldgp63 in Leishmania. Our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N significantly inhibits the secretion of secretory acid phosphatase by Leishmania. We have also found that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N retains RFP-Ldgp63 in Golgi and blocks the secretion of Ldgp63, whereas the trafficking of RFP-Ldgp63 in GFP-LdRab1:WT-expressing cells is unaltered in comparison with control cells. Taken together, our results have shown that the Rab1-regulated secretory pathway is well conserved, and hemoglobin receptor trafficking follows an Rab1-independent secretory pathway in Leishmania. PMID:26499792

  15. Effect of composition of titanium in silver-copper-titanium braze alloy on dissimilar laser brazing of binder-less cubic boron nitride and tungsten carbide

    NASA Astrophysics Data System (ADS)

    Sechi, Yoshihisa; Nagatsuka, Kimiaki; Nakata, Kazuhiro

    2014-08-01

    Laser brazing with Ti as an active element in silver-copper alloy braze metal has been carried out for binder-less cubic boron nitride and tungsten carbide, using silver-copper- titanium braze alloys with titanium content that varied between 0.28 mass% and 1.68 mass%. Observations of the interface using electron probe microanalysis and scanning acoustic microscopy show that efficient interface adhesion between binder-less cubic boron nitride and the silver-copper-titanium braze alloy was achieved for the braze with a titanium content of 0. 28 mass%.

  16. Munc13-4 Is a Rab11-binding Protein That Regulates Rab11-positive Vesicle Trafficking and Docking at the Plasma Membrane*

    PubMed Central

    Johnson, Jennifer L.; He, Jing; Ramadass, Mahalakshmi; Pestonjamasp, Kersi; Kiosses, William B.; Zhang, Jinzhong; Catz, Sergio D.

    2016-01-01

    The small GTPase Rab11 and its effectors control trafficking of recycling endosomes, receptor replenishment and the up-regulation of adhesion and adaptor molecules at the plasma membrane. Despite recent advances in the understanding of Rab11-regulated mechanisms, the final steps mediating docking and fusion of Rab11-positive vesicles at the plasma membrane are not fully understood. Munc13-4 is a docking factor proposed to regulate fusion through interactions with SNAREs. In hematopoietic cells, including neutrophils, Munc13-4 regulates exocytosis in a Rab27a-dependent manner, but its possible regulation of other GTPases has not been explored in detail. Here, we show that Munc13-4 binds to Rab11 and regulates the trafficking of Rab11-containing vesicles. Using a novel Time-resolved Fluorescence Resonance Energy Transfer (TR-FRET) assay, we demonstrate that Munc13-4 binds to Rab11a but not to dominant negative Rab11a. Immunoprecipitation analysis confirmed the specificity of the interaction between Munc13-4 and Rab11, and super-resolution microscopy studies support the interaction of endogenous Munc13-4 with Rab11 at the single molecule level in neutrophils. Vesicular dynamic analysis shows the common spatio-temporal distribution of Munc13-4 and Rab11, while expression of a calcium binding-deficient mutant of Munc13-4 significantly affected Rab11 trafficking. Munc13-4-deficient neutrophils showed normal endocytosis, but the trafficking, up-regulation, and retention of Rab11-positive vesicles at the plasma membrane was significantly impaired. This correlated with deficient NADPH oxidase activation at the plasma membrane in response to Rab11 interference. Our data demonstrate that Munc13-4 is a Rab11-binding partner that regulates the final steps of Rab11-positive vesicle docking at the plasma membrane. PMID:26637356

  17. Munc13-4 Is a Rab11-binding Protein That Regulates Rab11-positive Vesicle Trafficking and Docking at the Plasma Membrane.

    PubMed

    Johnson, Jennifer L; He, Jing; Ramadass, Mahalakshmi; Pestonjamasp, Kersi; Kiosses, William B; Zhang, Jinzhong; Catz, Sergio D

    2016-02-12

    The small GTPase Rab11 and its effectors control trafficking of recycling endosomes, receptor replenishment and the up-regulation of adhesion and adaptor molecules at the plasma membrane. Despite recent advances in the understanding of Rab11-regulated mechanisms, the final steps mediating docking and fusion of Rab11-positive vesicles at the plasma membrane are not fully understood. Munc13-4 is a docking factor proposed to regulate fusion through interactions with SNAREs. In hematopoietic cells, including neutrophils, Munc13-4 regulates exocytosis in a Rab27a-dependent manner, but its possible regulation of other GTPases has not been explored in detail. Here, we show that Munc13-4 binds to Rab11 and regulates the trafficking of Rab11-containing vesicles. Using a novel Time-resolved Fluorescence Resonance Energy Transfer (TR-FRET) assay, we demonstrate that Munc13-4 binds to Rab11a but not to dominant negative Rab11a. Immunoprecipitation analysis confirmed the specificity of the interaction between Munc13-4 and Rab11, and super-resolution microscopy studies support the interaction of endogenous Munc13-4 with Rab11 at the single molecule level in neutrophils. Vesicular dynamic analysis shows the common spatio-temporal distribution of Munc13-4 and Rab11, while expression of a calcium binding-deficient mutant of Munc13-4 significantly affected Rab11 trafficking. Munc13-4-deficient neutrophils showed normal endocytosis, but the trafficking, up-regulation, and retention of Rab11-positive vesicles at the plasma membrane was significantly impaired. This correlated with deficient NADPH oxidase activation at the plasma membrane in response to Rab11 interference. Our data demonstrate that Munc13-4 is a Rab11-binding partner that regulates the final steps of Rab11-positive vesicle docking at the plasma membrane.

  18. A new low-melting-point aluminum braze

    SciTech Connect

    Jacobson, D.M.; Humpston, G.; Sangha, S.P.S.

    1996-08-01

    Most high-strength aluminum engineering alloys cannot be joined by brazing because they either degrade or melt at the temperature at which commercially available aluminum brazes are used. Previous efforts to develop aluminum brazing filler metal alloys with a significantly reduced melting point have tended to be frustrated by poor mechanical properties of the alloys, corrosion of the joints or the high cost, toxicity or volatility of the constituent materials. This paper describes the development and assessment of a new brazing alloy with a composition of 73Al-20Cu-2Ni-55I (wt-%), which has been designed to overcome these limitations. A joining process has been devised for fluxless brazing of aluminum engineering alloys using the new filler metal for use in both inert gas and vacuum furnaces. The production of ductile foil preforms and roll-clad base metals is described together with preliminary results of mechanical property assessments and corrosion resistance trials. These results are highly encouraging and point to promising new applications for aluminum brazing technology.

  19. Microstructural and Mechanical Characterization of Actively Brazed Alumina Specimens

    SciTech Connect

    Hosking, F.M.; Cadden, C.H.; Stephens, J.J.; Glass, S.J.; Yang, N.Y.C.; Vianco, P.V.; Walker, C.A.

    1999-08-26

    Alumina (94 and 99.8% grade compositions) was brazed directly to itself with gold-based active brazing alloys (ABA's) containing vanadium additions of 1,2 and 3 weight percent. The effects of brazing conditions on the joint properties were investigated. Wetting behavior, interfacial reactions, microstructure, hermeticity and tensile strength were determined. Wetting was fair to good for the ABA and base material combinations. Microanalysis identified a discontinuous Al-V-O spinel reaction product at the alumina-braze interface. Tensile strength results for 94% alumina were uniformly good and generally not sensitive to the vanadium concentration, with tensile values of 85-105 MPa. There was more variability in the 99.8% alumina strength results, with values ranging from 25-95 MPa. The highest vanadium concentration (3 wt. %) yielded the highest joint strength for the brazed 99.8% alumina. Failures in the 99.8% alumina samples occurred at the braze-alumina interface, while the 94% alumina specimens exhibited fracture of the ceramic substrate.

  20. Reducing inadvertent alloying of metal/ceramic brazes

    SciTech Connect

    Stephens, J.J.; Hlava, P.F.

    1992-12-31

    Inadvertent alloying of Cu braze metal can compromise metal/ceramic seals. Electron microprobe analyses have quantified alloying of Cu brazes in metal/ceramic feedthroughs. Pin material and processing parameters above 1084C both affect alloying levels. Using either Kovar or Ni-plated 316L stainless steel pins limits alloying compared to Palco pins. Minimizing the time during which the braze is molten also avoids excessive alloying. The original thickness of the Ni plating on the Mo-Mn metallization of the ceramic also influences the alloying content of these brazes. Metal/ceramic brazes made with long brazing cycles, Mo-Mn metallization, and Kovar components grow a layer of Mo{sub 6}(Fe{sub 3.5}CO{sub 3.5}){sub 7} on the metallization. Layer thicknesses observed do not appear to compromise joint integrity. Ni additions of approximately 10 and 20 wt.% to Cu apparently increases the stress required for stress relaxation during cooldown. to maintain creep rates required for stress relaxation during cooldown. Relative to unalloyed Cu, this strengthening effect tends to increase as temperature is decreased.

  1. Brazing of titanium-vapor-coated silicon nitride

    SciTech Connect

    Santella, M.L. )

    1988-09-01

    A technique for brazing Si{sub 3}N{sub 4} with metallic alloys was evaluated. The process involved vapor coating the ceramic with a 1.0-{mu}-thick layer of titanium before the brazing operation. The coating improved wetting of the Si{sub 3}N{sub 4} surfaces to the extent that strong bonding between the solidified braze filler metal and the ceramic occurred. Braze joints of Si{sub 3}N{sub 4} were made with Ag-Cu, Au-Ni, and Au-Ni-Pd alloys at temperatures of 790{degree}, 970{degree}, and 1,130{degree}C. Silicon nitride specimens were also brazed with a Ag-Cu alloy to the molybdenum alloy TZM, titanium, and A286 steel at 790{degree}C. Residual stresses resulting from mismatch of thermal expansion coefficients between the Si{sub 3}N{sub 4} and the metals caused all of the ceramic-to-metal joints to spontaneously crack in the Si{sub 3}N{sub 4} upon cooling from the brazing temperature.

  2. Electron Beam Welder Used to Braze Sapphire to Platinum

    NASA Technical Reports Server (NTRS)

    Forsgren, Roger C.; Vannuyen, Thomas

    1998-01-01

    A new use for electron beam brazing was recently developed by NASA Lewis Research Center's Manufacturing Engineering Division. This work was done to fabricate a fiberoptic probe (developed by Sentec Corporation) that could measure high temperatures less than 600 deg C of vibrating machinery, such as in jet engine combustion research. Under normal circumstances, a sapphire fiber would be attached to platinum by a ceramic epoxy. However, no epoxies can adhere ceramic fibers to platinum under such high temperatures and vibration. Also, since sapphire and platinum have different thermal properties, the epoxy bond is subjected to creep over time. Therefore, a new method had to be developed that would permanently and reliably attach a sapphire fiber to platinum. Brazing a sapphire fiber to a platinum shell. The fiber-optic probe assembly consists of a 0.015-in.-diameter sapphire fiber attached to a 0.25-in.-long, 0.059-in.-diameter platinum shell. Because of the small size of this assembly, electron beam brazing was chosen instead of conventional vacuum brazing. The advantage of the electron beam is that it can generate a localized heat source in a vacuum. Gold reactive braze was used to join the sapphire fiber and the platinum. Consequently, the sapphire fiber was not affected by the total heat needed to braze the components together.

  3. Preliminary Process Design of ITER ELM Coil Bracket Brazing

    NASA Astrophysics Data System (ADS)

    LI, Xiangbin; SHI, Yi

    2015-03-01

    With the technical requirement of the International Thermonuclear Experimental Reactor (ITER) project, the manufacture and assembly technology of the mid Edge Localized Modes (ELM) coil was developed by the Institute of Plasma Physics, Chinese Academy of Science (ASIPP). As the gap between the bracket and the Stainless Steel jacketed and Mineral Insulated Conductor (SSMIC) can be larger than 0.5 mm instead of 0.01 mm to 0.1 mm as in normal industrial cases, the process of mid ELM coil bracket brazing to the SSMICT becomes quiet challenging, from a technical viewpoint. This paper described the preliminary design of ELM coil bracket brazing to the SSMIC process, the optimal bracket brazing curve and the thermal simulation of the bracket furnace brazing method developed by ANSYS. BAg-6 foil (Bag50Cu34Zn16) plus BAg-1a paste (Bag45CuZnCd) solders were chosen as the brazing filler. By testing an SSMICT prototype, it is shown that the average gap between the bracket and the SSMIC could be controlled to 0.2-0.3 mm, and that there were few voids in the brazing surface. The results also verified that the preliminary design had a favorable heat conducting performance in the bracket.

  4. Sawing performance comparison of brazed and sintered diamond wires

    NASA Astrophysics Data System (ADS)

    Huang, Guoqin; Xu, Xipeng

    2013-03-01

    Great attention has been paid on fabricating diamond wire by using the brazing diamond because of its strong chemical bonding strength and controllability of grits distribution. Although several serving performances of brazed diamond wire have been reported, seldom do these studies refer to its process characteristics. Sawing performances of a brazed diamond wire are investigated and compared with those of a sintered diamond wire on a wire saw machine. The surface topographies of beads selected from the two wires are micro observed before sawing. The sawing tests are carried out in constant feed rate feeding(CFF) and constant normal force feeding(CNFF). In CFF test, sawing force, power, and the cut depths of positions on contact curve are measured. Then, coupled with the observations of beads topographies, sawing force and its ratio, relations of power against material removal rate, and contact curve linearity are compared and discussed. In CNFF test, the sawing rates of the two wires are investigated. The results indicate that the brazed wire performs with lower sawing force(less 16% of tangential force and 28% of normal force), more energy efficiency(nearly one-fifth of sawing power is saved), at a higher sawing rate (the rate is doubled) and with better contact curve linearity as compared with the sintered wire. This proposed research experimentally evaluates the sawing performances of brazed diamond wire from the aspect of process parameters, which can provide a basis for popularizing the brazed diamond wire.

  5. Environmental simulation evaluation of SSiC brazed optical mirrors

    NASA Astrophysics Data System (ADS)

    Liu, Yan; Ma, Zhen; Chen, Jian; Chen, Zhongming; Liu, Xuejian; Huang, Zhengren

    2014-09-01

    Sintered silicon carbide (SSiC) is becoming one of the most important materials for the optical mirrors due to its excellent specific stiffness (E/ρ) and demission stability (λ/α). However, it is difficult to fabricate the monolithic structure SSiC optical mirror with demission of larger than Φ1.5m because of process limitation. Joining of SSiC segments (brazing) provide a good solution to prepare large size mirror optics. However, compared with the uniform properties of the monolithic structure SSiC optical mirror, the brazed mirror is composed of two materials (SSiC segments and brazing material), so the performance of optical grinding and reliability of brazed optical mirrors become the focus. In this paper, the Φ300mm and Φ600mm brazed optical mirrors was used to evaluate the reliability of different conditions. Three kinds of environmental simulation tests, including thermal stability, thermal circle and random vibration were carried out. The evaluation results show that the temperature and vibration has no obvious effects on the surface figure (RMS) of the brazed optical mirrors.

  6. Effects induced by the brazing thermal cycle on the structural properties of materials in steel brazed joints

    NASA Astrophysics Data System (ADS)

    Brossa, M.; Guerreschi, U.

    1988-07-01

    Specimens of AISI 316 austenitic steel, 9Cr-1Mo modified martensitic steel, and 2.25Cr-1Mo ferritic steel have been brazed in a furnace under vacuum conditions. Several thermal cycles were followed in order to investigate their influence on the final characteristics of the joints and the materials. Various kinds of high-melting Ni-based brazing alloys having properties similar to the base materials were used. The growth of the austenitic grain size was measured in order to assess its dependence on the brazing cycle of the different alloys. Metallurgical analyses were carried out and, in a few cases, repeated after the specimens underwent a second thermal cycle or destructive tests. No precipitation of carbides was observed in the base materials after the brazing cycles, confirming that the cooling rate is above the critical range.

  7. Brazing open cell reticulated copper foam to stainless steel tubing with vacuum furnace brazed gold/indium alloy plating

    DOEpatents

    Howard, Stanley R.; Korinko, Paul S.

    2008-05-27

    A method of fabricating a heat exchanger includes brush electroplating plated layers for a brazing alloy onto a stainless steel tube in thin layers, over a nickel strike having a 1.3 .mu.m thickness. The resultant Au-18 In composition may be applied as a first layer of indium, 1.47 .mu.m thick, and a second layer of gold, 2.54 .mu.m thick. The order of plating helps control brazing erosion. Excessive amounts of brazing material are avoided by controlling the electroplating process. The reticulated copper foam rings are interference fit to the stainless steel tube, and in contact with the plated layers. The copper foam rings, the plated layers for brazing alloy, and the stainless steel tube are heated and cooled in a vacuum furnace at controlled rates, forming a bond of the copper foam rings to the stainless steel tube that improves heat transfer between the tube and the copper foam.

  8. Brazing of Stainless Steel to Yttria-Stabilized Zirconia Using Gold-Based Brazes for Solid Oxide Fuel Cell Applications

    NASA Technical Reports Server (NTRS)

    Singh, M.; Shpargel, T. P.; Asthana, R.

    2007-01-01

    Two gold-base active metal brazes (gold-ABA and gold-ABA-V) were evaluated for oxidation resistance to 850 C, and used to join yttria-stabilized zirconia (YSZ) to a corrosion-resistant ferritic stainless steel for possible use in solid oxide fuel cells. Thermogravimetric analysis and optical microscopy and scanning electron microscopy coupled with energy-dispersive spectroscopy were used to evaluate the braze oxidation behavior, and microstructure and composition of the YSZ/braze/steel joints. Both gold-ABA and gold-ABA-V exhibited nearly linear oxidation kinetics at 850 C, with gold-ABA-V showing faster oxidation than gold-ABA. Both brazes produced metallurgically sound YSZ/steel joints due to chemical interactions of Ti and V with the YSZ and steel substrates.

  9. Method For Brazing And Thermal Processing

    DOEpatents

    Milewski, John O.; Dave, Vivek R.; Christensen, Dane; Carpenter, II, Robert W.

    2005-07-12

    The present invention includes a method for brazing of two objects or heat treatment of one object. First, object or objects to be treated are selected and initial conditions establishing a relative geometry and material characteristics are determined. Then, a first design of an optical system for directing heat energy onto the object or objects is determined. The initial conditions and first design of the optical system are then input into a optical ray-tracing computer program. The program is then run to produce a representative output of the heat energy input distribution to the object or objects. The geometry of the object or objects, material characteristics, and optical system design are then adjusted until an desired heat input is determined.

  10. Cloning and characterization of ApRab4, a recycling Rab protein of Aiptasia pulchella, and its implication in the symbiosome biogenesis.

    PubMed

    Hong, Ming-Cheng; Huang, Yung-Sen; Song, Pa-Ching; Lin, Wen-Wen; Fang, Lee-Shing; Chen, Ming-Chyuan

    2009-01-01

    The biogenesis of Symbiodinium symbiosome in the host cells of the sea anemone, Aiptasia pulchella, involves retention of ApRab5 on and exclusion of ApRab11 from the organelle. One predicted consequence of this differential Rab association is the constant membrane fusion of symbiosomes with endocytic vesicles in the absence of parallel membrane retrieval and the subsequent formation of spacious symbiosomes, which nevertheless, contradicts the common perception. To solve this discrepancy, we determined whether membrane fusion occurs between symbiosomes and endocytic vesicles, and whether ApRab11-independent recycling is involved in symbiosome biogenesis. By using the biotin-avidin detection system, we found evidence for symbiosome-endocytic vesicle fusion. Cloning and characterization of ApRab4, an A. pulchella homolog of Rab4, showed that ApRab4 is associated with both the early endocytic and the perinuclear recycling compartments, and its normal function is required for the organization of the recycling compartments. Immunostaining localized ApRab4 to the symbiosome membrane, partially overlapping with ApRab5-decorated microdomains. Significantly, a treatment that impaired Symbiodinium photosynthesis also abolished symbiosome association of ApRab4. Furthermore, ApRab4 was quickly recruited to newly formed phagosomes, but prolonged association only occurred in those harboring live zooxanthelllae. We propose that ApRab4 retention on the symbiosome is an essential part of the mechanism for the biogenesis of Symbiodinium symbiosome.

  11. 32 CFR 202.4 - Composition of a RAB.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... appropriate. At closing installations where BRAC Cleanup Teams (BCT) exist, representatives of the BCT may.... Community RAB members should live and/or work in the affected community or be affected by the...

  12. 32 CFR 202.4 - Composition of a RAB.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... appropriate. At closing installations where BRAC Cleanup Teams (BCT) exist, representatives of the BCT may.... Community RAB members should live and/or work in the affected community or be affected by the...

  13. Direct targeting of Rab-GTPase-effector interactions.

    PubMed

    Spiegel, Jochen; Cromm, Philipp M; Itzen, Aymelt; Goody, Roger S; Grossmann, Tom N; Waldmann, Herbert

    2014-02-24

    Small GTPases are molecular switches using GDP/GTP alternation to control numerous vital cellular processes. Although aberrant function and regulation of GTPases are implicated in various human diseases, direct targeting of this class of proteins has proven difficult, as GTPase signaling and regulation is mediated by extensive and shallow protein interfaces. Here we report the development of inhibitors of protein-protein interactions involving Rab proteins, a subfamily of GTPases, which are key regulators of vesicular transport. Hydrocarbon-stapled peptides were designed based on crystal structures of Rab proteins bound to their interaction partners. These modified peptides exhibit significantly increased affinities and include a stapled peptide (StRIP3) that selectively binds to activated Rab8a and inhibits a Rab8a-effector interaction in vitro.

  14. Rab8A regulates insulin-stimulated GLUT4 translocation in C2C12 myoblasts.

    PubMed

    Li, Hanbing; Ou, Liting; Fan, Jiannan; Xiao, Mei; Kuang, Cuifang; Liu, Xu; Sun, Yonghong; Xu, Yingke

    2017-02-01

    Rab proteins are important regulators of GLUT4 trafficking in muscle and adipose cells. It is still unclear which Rabs are involved in insulin-stimulated GLUT4 translocation in C2C12 myoblasts. In this study, we detect the colocalization of Rab8A with GLUT4 and the presence of Rab8A at vesicle exocytic sites by TIRFM imaging. Overexpression of dominant-negative Rab8A (T22N) diminishes insulin-stimulated GLUT4 translocation, while constitutively active Rab8A (Q67L) augments it. In addition, knockdown of Rab8A inhibits insulin-stimulated GLUT4 translocation, which is rescued by replenishment of RNAi-resistant Rab8A. Together, these results indicate an indispensable role for Rab8A in insulin-regulated GLUT4 trafficking in C2C12 cells.

  15. LIFE CYCLE DESIGN OF AIR INTAKE MANIFOLDS; PHASE I: 2.0 L FORD CONTOUR AIR INTAKE MANIFOLD

    EPA Science Inventory

    The project team applied the life cycle design methodology to the design analysis of three alternative air intake manifolds: a sand cast aluminum, brazed aluminum tubular, and nylon composite. The design analysis included a life cycle inventory analysis, environmental regulatory...

  16. α1B-Adrenergic Receptors Differentially Associate with Rab Proteins during Homologous and Heterologous Desensitization

    PubMed Central

    Castillo-Badillo, Jean A.; Sánchez-Reyes, Omar B.; Alfonzo-Méndez, Marco A.; Romero-Ávila, M. Teresa; Reyes-Cruz, Guadalupe; García-Sáinz, J. Adolfo

    2015-01-01

    Internalization of G protein-coupled receptors can be triggered by agonists or by other stimuli. The process begins within seconds of cell activation and contributes to receptor desensitization. The Rab GTPase family controls endocytosis, vesicular trafficking, and endosomal fusion. Among their remarkable properties is the differential distribution of its members on the surface of various organelles. In the endocytic pathway, Rab 5 controls traffic from the plasma membrane to early endosomes, whereas Rab 4 and Rab 11 regulate rapid and slow recycling from early endosomes to the plasma membrane, respectively. Moreover, Rab 7 and Rab 9 regulate the traffic from late endosomes to lysosomes and recycling to the trans-Golgi. We explore the possibility that α1B-adrenergic receptor internalization induced by agonists (homologous) and by unrelated stimuli (heterologous) could involve different Rab proteins. This possibility was explored by Fluorescence Resonance Energy Transfer (FRET) using cells coexpressing α1B-adrenergic receptors tagged with the red fluorescent protein, DsRed, and different Rab proteins tagged with the green fluorescent protein. It was observed that when α1B-adrenergic receptors were stimulated with noradrenaline, the receptors interacted with proteins present in early endosomes, such as the early endosomes antigen 1, Rab 5, Rab 4, and Rab 11 but not with late endosome markers, such as Rab 9 and Rab 7. In contrast, sphingosine 1-phosphate stimulation induced rapid and transient α1B-adrenergic receptor interaction of relatively small magnitude with Rab 5 and a more pronounced and sustained one with Rab 9; interaction was also observed with Rab 7. Moreover, the GTPase activity of the Rab proteins appears to be required because no FRET was observed when dominant-negative Rab mutants were employed. These data indicate that α1B-adrenergic receptors are directed to different endocytic vesicles depending on the desensitization type (homologous vs

  17. RAB-10-Dependent Membrane Transport Is Required for Dendrite Arborization.

    PubMed

    Zou, Wei; Yadav, Smita; DeVault, Laura; Nung Jan, Yuh; Sherwood, David R

    2015-01-01

    Formation of elaborately branched dendrites is necessary for the proper input and connectivity of many sensory neurons. Previous studies have revealed that dendritic growth relies heavily on ER-to-Golgi transport, Golgi outposts and endocytic recycling. How new membrane and associated cargo is delivered from the secretory and endosomal compartments to sites of active dendritic growth, however, remains unknown. Using a candidate-based genetic screen in C. elegans, we have identified the small GTPase RAB-10 as a key regulator of membrane trafficking during dendrite morphogenesis. Loss of rab-10 severely reduced proximal dendritic arborization in the multi-dendritic PVD neuron. RAB-10 acts cell-autonomously in the PVD neuron and localizes to the Golgi and early endosomes. Loss of function mutations of the exocyst complex components exoc-8 and sec-8, which regulate tethering, docking and fusion of transport vesicles at the plasma membrane, also caused proximal dendritic arborization defects and led to the accumulation of intracellular RAB-10 vesicles. In rab-10 and exoc-8 mutants, the trans-membrane proteins DMA-1 and HPO-30, which promote PVD dendrite stabilization and branching, no longer localized strongly to the proximal dendritic membranes and instead were sequestered within intracellular vesicles. Together these results suggest a crucial role for the Rab10 GTPase and the exocyst complex in controlling membrane transport from the secretory and/or endosomal compartments that is required for dendritic growth.

  18. Rab proteins: the key regulators of intracellular vesicle transport.

    PubMed

    Bhuin, Tanmay; Roy, Jagat Kumar

    2014-10-15

    Vesicular/membrane trafficking essentially regulates the compartmentalization and abundance of proteins within the cells and contributes in many signalling pathways. This membrane transport in eukaryotic cells is a complex process regulated by a large and diverse array of proteins. A large group of monomeric small GTPases; the Rabs are essential components of this membrane trafficking route. Most of the Rabs are ubiquitously expressed proteins and have been implicated in vesicle formation, vesicle motility/delivery along cytoskeleton elements and docking/fusion at target membranes through the recruitment of effectors. Functional impairments of Rabs affecting transport pathways manifest different diseases. Rab functions are accompanied by cyclical activation and inactivation of GTP-bound and GDP-bound forms between the cytosol and membranes which is regulated by upstream regulators. Rab proteins are characterized by their distinct sub-cellular localization and regulate a wide variety of endocytic, transcytic and exocytic transport pathways. Mutations of Rabs affect cell growth, motility and other biological processes.

  19. RAB-10-Dependent Membrane Transport Is Required for Dendrite Arborization

    PubMed Central

    Zou, Wei; Yadav, Smita; DeVault, Laura; Jan, Yuh Nung; Sherwood, David R.

    2015-01-01

    Formation of elaborately branched dendrites is necessary for the proper input and connectivity of many sensory neurons. Previous studies have revealed that dendritic growth relies heavily on ER-to-Golgi transport, Golgi outposts and endocytic recycling. How new membrane and associated cargo is delivered from the secretory and endosomal compartments to sites of active dendritic growth, however, remains unknown. Using a candidate-based genetic screen in C. elegans, we have identified the small GTPase RAB-10 as a key regulator of membrane trafficking during dendrite morphogenesis. Loss of rab-10 severely reduced proximal dendritic arborization in the multi-dendritic PVD neuron. RAB-10 acts cell-autonomously in the PVD neuron and localizes to the Golgi and early endosomes. Loss of function mutations of the exocyst complex components exoc-8 and sec-8, which regulate tethering, docking and fusion of transport vesicles at the plasma membrane, also caused proximal dendritic arborization defects and led to the accumulation of intracellular RAB-10 vesicles. In rab-10 and exoc-8 mutants, the trans-membrane proteins DMA-1 and HPO-30, which promote PVD dendrite stabilization and branching, no longer localized strongly to the proximal dendritic membranes and instead were sequestered within intracellular vesicles. Together these results suggest a crucial role for the Rab10 GTPase and the exocyst complex in controlling membrane transport from the secretory and/or endosomal compartments that is required for dendritic growth. PMID:26394140

  20. Retort braze bonding of borsic/aluminum composite sheet to titanium

    NASA Technical Reports Server (NTRS)

    Webb, B. A.; Dolowy, J. F., Jr.

    1975-01-01

    Braze bonding studies between Borsic/aluminum composite and titanium sheet were conducted to establish acceptable brazing techniques and to assess potential joint efficiencies. Excellent braze joints were produced which exhibited joint strengths exceeding 117 MPa (17,000 psi) and which retained up to 2/3 of this strength at 589 K (600 F). Noticeable composite strength degradation resulting from the required high temperature braze cycle was found to be a problem.

  1. Inert-gas welding and brazing enclosure fabricated from sheet plastic

    NASA Technical Reports Server (NTRS)

    Wisner, J. P.

    1965-01-01

    Custom-fabricated plastic bag maintains an inert-gas atmosphere for welding and brazing certain metals. The bag fits over part of the workpieces and the welding and brazing tools. It is also used for metal brazing and fusion plating which require an inert-gas atmosphere.

  2. 49 CFR 178.50 - Specification 4B welded or brazed steel cylinders.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 3 2013-10-01 2013-10-01 false Specification 4B welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.50 Specification 4B welded or brazed steel cylinders. (a) Type, size, and service pressure. A DOT 4B is a welded or brazed steel cylinder with...

  3. 49 CFR 178.51 - Specification 4BA welded or brazed steel cylinders.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 3 2012-10-01 2012-10-01 false Specification 4BA welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.51 Specification 4BA welded or brazed steel...) Cylindrical type cylinders must be of circumferentially welded or brazed construction. (b) Steel. The...

  4. 49 CFR 178.51 - Specification 4BA welded or brazed steel cylinders.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 3 2013-10-01 2013-10-01 false Specification 4BA welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.51 Specification 4BA welded or brazed steel...) Cylindrical type cylinders must be of circumferentially welded or brazed construction. (b) Steel. The...

  5. 49 CFR 178.51 - Specification 4BA welded or brazed steel cylinders.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 3 2014-10-01 2014-10-01 false Specification 4BA welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.51 Specification 4BA welded or brazed steel...) Cylindrical type cylinders must be of circumferentially welded or brazed construction. (b) Steel. The...

  6. 49 CFR 178.50 - Specification 4B welded or brazed steel cylinders.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 3 2011-10-01 2011-10-01 false Specification 4B welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.50 Specification 4B welded or brazed steel cylinders. (a) Type, size, and service pressure. A DOT 4B is a welded or brazed steel cylinder with...

  7. 49 CFR 178.50 - Specification 4B welded or brazed steel cylinders.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 3 2014-10-01 2014-10-01 false Specification 4B welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.50 Specification 4B welded or brazed steel cylinders. (a) Type, size, and service pressure. A DOT 4B is a welded or brazed steel cylinder with...

  8. 49 CFR 178.51 - Specification 4BA welded or brazed steel cylinders.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 3 2011-10-01 2011-10-01 false Specification 4BA welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.51 Specification 4BA welded or brazed steel...) Cylindrical type cylinders must be of circumferentially welded or brazed construction. (b) Steel. The...

  9. 49 CFR 178.50 - Specification 4B welded or brazed steel cylinders.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 3 2012-10-01 2012-10-01 false Specification 4B welded or brazed steel cylinders... FOR PACKAGINGS Specifications for Cylinders § 178.50 Specification 4B welded or brazed steel cylinders. (a) Type, size, and service pressure. A DOT 4B is a welded or brazed steel cylinder with...

  10. Substrate Effects on the High Temperature Oxidation Behavior of a Gold-Based Braze Filler Metal

    SciTech Connect

    Weil, K. Scott; Rice, Joseph P.

    2005-06-30

    Oxidation testing was conducted on a commercial gold-based braze alloy, Gold ABA, and on zirconia and stainless steel joining couples prepared using this braze filler metal. Preliminary results reveal that both substrates play a significant role in determining the overall oxidation resistance of the brazed joint.

  11. Rab27b regulates number and secretion of platelet dense granules.

    PubMed

    Tolmachova, Tanya; Abrink, Magnus; Futter, Clare E; Authi, Kalwant S; Seabra, Miguel C

    2007-04-03

    The Rab27 GTPase subfamily consists of two closely related homologs, Rab27a and Rab27b. Rab27a has been shown previously to regulate organelle movement and regulated exocytosis in a wide variety of secretory cells. However, the role of the more restrictedly expressed Rab27b remains unclear. Here we describe the creation of Rab27b knockout (KO) strain that was subsequently crossed with the naturally occurring Rab27a KO line, ashen, to produce double KO (Rab27a(ash/ash) Rab27b(-/-)) mice. Rab27b KO (and double KO) exhibit significant hemorrhagic disease in contrast to ashen mice. In vitro assays demonstrated impaired aggregation with collagen and U46619 and reduced secretion of dense granules in both Rab27b and double KO strains. Additionally, we detected a 50% reduction in the number of dense granules per platelet and diminished platelet serotonin content, possibly due to a dense granule packaging defect into proplatelets during megakaryocyte maturation. The presence of Rab27a partially compensated for the secretory defect but not the reduced granule number. The morphology and function of platelet alpha-granules were unaffected. Our data suggest that Rab27b is a key regulator of dense granule secretion in platelets and thus a candidate gene for delta-storage pool deficiency in humans.

  12. Small GTPase Rab40c Associates with Lipid Droplets and Modulates the Biogenesis of Lipid Droplets

    PubMed Central

    Tan, Ran; Wang, Weijie; Wang, Shicong; Wang, Zhen; Sun, Lixiang; He, Wei; Fan, Rong; Zhou, Yunhe; Xu, Xiaohui; Hong, Wanjin; Wang, Tuanlao

    2013-01-01

    The subcellular location and cell biological function of small GTPase Rab40c in mammalian cells have not been investigated in detail. In this study, we demonstrated that the exogenously expressed GFP-Rab40c associates with lipid droplets marked by neutral lipid specific dye Oil red or Nile red, but not with the Golgi or endosomal markers. Further examination demonstrated that Rab40c is also associated with ERGIC-53 containing structures, especially under the serum starvation condition. Rab40c is increasingly recruited to the surface of lipid droplets during lipid droplets formation and maturation in HepG2 cells. Rab40c knockdown moderately decreases the size of lipid droplets, suggesting that Rab40c is involved in the biogenesis of lipid droplets. Stimulation for adipocyte differentiation increases the expression of Rab40c in 3T3-L1 cells. Rab40c interacts with TIP47, and is appositionally associated with TIP47-labeled lipid droplets. In addition, over-expression of Rab40c causes the clustering of lipid droplets independent of its GTPase activity, but completely dependent of the intact SOCS box domain of Rab40c. In addition, Rab40c displayed self-interaction as well as interaction with TIP47 and the SOCS box is essential for its ability to induce clustering of lipid droplets. Our results suggest that Rab40c is a novel Rab protein associated with lipid droplets, and is likely involved in modulating the biogenesis of lipid droplets. PMID:23638186

  13. Real-time imaging of Rab5 activity using a prequenched biosensor.

    PubMed

    Zhan, Ke; Xie, Hexin; Gall, Jessica; Ma, Manlung; Griesbeck, Oliver; Salehi, Ahmad; Rao, Jianghong

    2011-07-15

    A key regulator of receptor-mediated endocytosis, Rab5, plays a pivotal role in cargo receptor internalization, endosomal maturation, and transduction and degradation of internalized signaling molecules and recycling cargo receptor. Stressful conditions within cells lead to increased Rab5 activation, and increasing evidence correlates Rab5 activity abnormalities with certain diseases. Current antibody-based imaging methods cannot distinguish active Rab5 from total Rab5 population and provide dynamic information on magnitude and duration of Rab5 activation in cellular events and pathogenesis. We report here novel molecular imaging probes that specifically target GTP-bound Rab5 associated with the early endosome membrane in live cells and fixed mouse brain tissues. Our Rab5 activity fluorescent biosensor (RAFB) contains the Rab5 binding domain of the Rab5 effector Rabaptin 5, a fluorophore (a quantum dot or fluorescent dye) and a cell-penetrating peptide for live-cell delivery. The quantum dot conjugated RAFB was able to image the elevated Rab5 activity in both the cortex and hippocampi tissues of a Ts65Dn mouse. A prequenched RAFB based on fluorescence resonance energy transfer (FRET) can image cytosolic active Rab5 in single live cells. This novel method should enable imaging of the biological process in which Rab5 activity is regulated in various cellular systems.

  14. Air

    MedlinePlus

    ... do to protect yourself from dirty air . Indoor air pollution and outdoor air pollution Air can be polluted indoors and it can ... this chart to see what things cause indoor air pollution and what things cause outdoor air pollution! Indoor ...

  15. Characterization of the molecular defects in Rab27a, caused by RAB27A missense mutations found in patients with Griscelli syndrome.

    PubMed

    Bahadoran, Philippe; Busca, Roser; Chiaverini, Christine; Westbroek, Wendy; Lambert, Jo; Bille, Karine; Valony, Gäelle; Fukuda, Mitsunori; Naeyaert, Jean-Marie; Ortonne, Jean-Paul; Ballotti, Robert

    2003-03-28

    Rab27a plays a pivotal role in the transport of melanosomes to dendrite tips of melanocytes and mutations in RAB27A, which impair melanosome transport cause the pigmentary dilution and the immune deficiency found in several patients with Griscelli syndrome (GS). Interestingly, three GS patients present single homozygous missense mutations in RAB27A, leading to W73G, L130P, and A152P transitions that affect highly conserved residues among Rab proteins. However, the functional consequences of these mutations have not been studied. In the present report, we evaluated the effect of overexpression of these mutants on melanosome, melanophilin, and myosin-Va localization in B16 melanoma cells. Then we studied several key parameters for Rab27a function, including GTP binding and interaction with melanophilin/myosin-Va complex, which links melanosomes to the actin network. Our results showed that Rab27a-L130P cannot bind GTP, does not interact with melanophilin, and consequently cannot allow melanosome transport on the actin filaments. Interestingly, Rab27a-W73G binds GTP but does not interact with melanophilin. Thus, Rab27a-W73G cannot support the actin-dependent melanosome transport. Finally, Rab27a-A152P binds both GTP and melanophilin. However, Rab27a-A152P does not allow melanosome transport and acts as a dominant negative mutant, because its overexpression, in B16 melanoma cells, mimics a GS phenotype. Hence, the interaction of Rab27a with melanophilin/myosin-Va is not sufficient to ensure a correct melanosome transport. Our results pointed to an unexpected complexity of Rab27a function and open the way to the search for new Rab27a effectors or regulators that control the transport of Rab27a-dependent vesicles.

  16. Direct use geothermal applications for brazed plate heat exchangers

    SciTech Connect

    Rafferty, K.

    1993-02-01

    Brazed plate heat exchanger were placed in three geothermal fluids (Klamath Falls, OR; Boise, ID; and Pagosa Springs, CO) in order to determine the effect of H{sub 2}S on braze material. Based on subsequent analysis, it appears that the rate of corrosion of the braze material is much slower than corrosion of copper tube materials in the same fluids. Minimum expected life of the heat exchangers based on these corrosion rates is reported to be 12 years in fluids of less than 1 ppm H{sub 2}S and 10 years in fluids of less than 5 ppm. Based on these expected lives, and using a 3% inflation rate and 8% discount rate, brazed plate heat exchangers are a clear economic choice in which the capital cost is 50% or less of the cost of a plate and frame heat exchanger for the same duty. Due to their single pass design, brazed plate heat exchangers are generally limited to approach temperatures of 10{degree} or greater. Size limitations restrict applications to 100 gpm and/or 200 ft{sup 2} heat transfer surface area.

  17. Direct use geothermal applications for brazed plate heat exchangers

    SciTech Connect

    Rafferty, K.

    1993-02-01

    Brazed plate heat exchanger were placed in three geothermal fluids (Klamath Falls, OR; Boise, ID; and Pagosa Springs, CO) in order to determine the effect of H[sub 2]S on braze material. Based on subsequent analysis, it appears that the rate of corrosion of the braze material is much slower than corrosion of copper tube materials in the same fluids. Minimum expected life of the heat exchangers based on these corrosion rates is reported to be 12 years in fluids of less than 1 ppm H[sub 2]S and 10 years in fluids of less than 5 ppm. Based on these expected lives, and using a 3% inflation rate and 8% discount rate, brazed plate heat exchangers are a clear economic choice in which the capital cost is 50% or less of the cost of a plate and frame heat exchanger for the same duty. Due to their single pass design, brazed plate heat exchangers are generally limited to approach temperatures of 10[degree] or greater. Size limitations restrict applications to 100 gpm and/or 200 ft[sup 2] heat transfer surface area.

  18. Direct use geothermal applications for brazed plate heat exchangers

    SciTech Connect

    Rafferty, K.

    1992-12-31

    Brazed plate heat exchanger were placed in three geothermal fluids (Klamath Falls, OR; Boise, ID; and Pagosa Springs, CO) to determine the effect of H{sub 2}S on braze material. Based on subsequent analysis, it appears that the rate of corrosion of the braze material is much slower than corrosion of copper tube materials in the same fluids. Minimum expected life of the heat exchangers based on these corrosion rates is reported to be 12 years in fluids of less than 1 ppm H{sub 2}S and 10 years in fluids of less than 5 ppm. Based on these expected lives, and using a 3% inflation rate and 8% discount rate, brazed plate heat exchangers are a clear economic choice in which the capital cost is 50% or less of the cost of a plate and frame heat exchanger for the same duty. Due to their single pass design, brazed plate heat exchangers are generally limited to approach temperatures of 10{degrees} or greater. Size limitations restrict applications to 100 gpm and/or 200 ft{sup 2} heat transfer surface area.

  19. Mechanical properties of high-temperature brazed titanium materials

    SciTech Connect

    Lugscheider, E.; Broich, U.

    1995-05-01

    The mechanical properties of commercial titanium CPTi and Ti-Al6-V4 joints, brazed with Ti-based filler metals in the system Ti(Zr)-Cu-Ni-(Pd) are evaluated by tensile test at various temperatures, as well as by fatigue test at room temperature. The influence of the microstructure in the brazing zone on the mechanical properties of the joints was assessed by conducting metallographic analysis. A vacuum furnace and an induction heating furnace were used for the production of the metallographic and tensile samples. The results from the mechanical and metallographic investigations revealed a strong dependence of the tensile strength of the titanium joints on the microstructure of the brazing zone. The presence of the brittle intermetallic Ti-Cu and Ti-Ni phases in the brazing zone leads to the weakening of the joint. However, for the formation of these intermetallic phases can be avoided by using adequate brazing process parameters and by optimizing the joint clearance. In that case, it is possible to fabricate titanium joints with Ti-based filler metals that have excellent mechanical properties comparable to those of the base metal.

  20. Fabrication of large aperture SiC brazing mirror

    NASA Astrophysics Data System (ADS)

    Li, Ang; Wang, Peipei; Dong, Huiwen; Wang, Peng

    2016-10-01

    The SiC brazing mirror is the mirror whose blank is made by assembling together smaller SiC pieces with brazing technique. Using such kinds of joining techniques, people can manufacture large and complex SiC assemblies. The key technologies of fabricating and testing SiC brazing flat mirror especially for large aperture were studied. The SiC brazing flat mirror was ground by smart ultrasonic-milling machine, and then it was lapped by the lapping smart robot and measured by Coordinate Measuring Machine (CMM). After the PV of the surface below 4um, we did classic coarse polishing to the surface and studied the shape of the polishing tool which directly effects removal amount distribution. Finally, it was figured by the polishing smart robot and measured by Fizeau interferometer. We also studied the influence of machining path and removal functions of smart robots on the manufacturing results and discussed the use of abrasive in this process. At last, an example for fabricating and measuring a similar SiC brazing flat mirror with the aperture of 600 mm made by Shanghai Institute of Ceramics was given. The mirror blank consists of 6 SiC sectors and the surface was finally processed to a result of the Peak-to-Valley (PV) 150nm and Root Mean Square (RMS) 12nm.

  1. Direct use geothermal applications for brazed plate heat exchangers

    SciTech Connect

    Rafferty, K.

    1992-01-01

    Brazed plate heat exchanger were placed in three geothermal fluids (Klamath Falls, OR; Boise, ID; and Pagosa Springs, CO) to determine the effect of H[sub 2]S on braze material. Based on subsequent analysis, it appears that the rate of corrosion of the braze material is much slower than corrosion of copper tube materials in the same fluids. Minimum expected life of the heat exchangers based on these corrosion rates is reported to be 12 years in fluids of less than 1 ppm H[sub 2]S and 10 years in fluids of less than 5 ppm. Based on these expected lives, and using a 3% inflation rate and 8% discount rate, brazed plate heat exchangers are a clear economic choice in which the capital cost is 50% or less of the cost of a plate and frame heat exchanger for the same duty. Due to their single pass design, brazed plate heat exchangers are generally limited to approach temperatures of 10[degrees] or greater. Size limitations restrict applications to 100 gpm and/or 200 ft[sup 2] heat transfer surface area.

  2. Mevalonate kinase deficiency leads to decreased prenylation of Rab GTPases

    PubMed Central

    Jurczyluk, Julie; Munoz, Marcia A; Skinner, Oliver P; Chai, Ryan C; Ali, Naveid; Palendira, Umaimainthan; Quinn, Julian MW; Preston, Alexandra; Tangye, Stuart G; Brown, Andrew J; Argent, Elizabeth; Ziegler, John B; Mehr, Sam; Rogers, Michael J

    2016-01-01

    Mevalonate kinase deficiency (MKD) is caused by mutations in a key enzyme of the mevalonate–cholesterol biosynthesis pathway, leading to recurrent autoinflammatory disease characterised by enhanced release of interleukin-1β (IL-1β). It is currently believed that the inflammatory phenotype of MKD is triggered by temperature-sensitive loss of mevalonate kinase activity and reduced biosynthesis of isoprenoid lipids required for the prenylation of small GTPase proteins. However, previous studies have not clearly shown any change in protein prenylation in patient cells under normal conditions. With lymphoblast cell lines from two compound heterozygous MKD patients, we used a highly sensitive in vitro prenylation assay, together with quantitative mass spectrometry, to reveal a subtle accumulation of unprenylated Rab GTPases in cells cultured for 3 days or more at 40 °C compared with 37 °C. This included a 200% increase in unprenylated Rab7A, Rab14 and Rab1A. Inhibition of sterol regulatory element-binding protein (SREBP) activation by fatostatin led to more pronounced accumulation of unprenylated Rab proteins in MKD cells but not parent cells, suggesting that cultured MKD cells may partially overcome the loss of isoprenoid lipids by SREBP-mediated upregulation of enzymes required for isoprenoid biosynthesis. Furthermore, while inhibition of Rho/Rac/Rap prenylation promoted the release of IL-1β, specific inhibition of Rab prenylation by NE10790 had no effect in human peripheral blood mononuclear cells or human THP-1 monocytic cells. These studies demonstrate for the first time that mutations in mevalonate kinase can lead to a mild, temperature-induced defect in the prenylation of small GTPases, but that loss of prenylated Rab GTPases is not the cause of enhanced IL-1β release in MKD. PMID:27377765

  3. Mevalonate kinase deficiency leads to decreased prenylation of Rab GTPases.

    PubMed

    Jurczyluk, Julie; Munoz, Marcia A; Skinner, Oliver P; Chai, Ryan C; Ali, Naveid; Palendira, Umaimainthan; Quinn, Julian Mw; Preston, Alexandra; Tangye, Stuart G; Brown, Andrew J; Argent, Elizabeth; Ziegler, John B; Mehr, Sam; Rogers, Michael J

    2016-11-01

    Mevalonate kinase deficiency (MKD) is caused by mutations in a key enzyme of the mevalonate-cholesterol biosynthesis pathway, leading to recurrent autoinflammatory disease characterised by enhanced release of interleukin-1β (IL-1β). It is currently believed that the inflammatory phenotype of MKD is triggered by temperature-sensitive loss of mevalonate kinase activity and reduced biosynthesis of isoprenoid lipids required for the prenylation of small GTPase proteins. However, previous studies have not clearly shown any change in protein prenylation in patient cells under normal conditions. With lymphoblast cell lines from two compound heterozygous MKD patients, we used a highly sensitive in vitro prenylation assay, together with quantitative mass spectrometry, to reveal a subtle accumulation of unprenylated Rab GTPases in cells cultured for 3 days or more at 40 °C compared with 37 °C. This included a 200% increase in unprenylated Rab7A, Rab14 and Rab1A. Inhibition of sterol regulatory element-binding protein (SREBP) activation by fatostatin led to more pronounced accumulation of unprenylated Rab proteins in MKD cells but not parent cells, suggesting that cultured MKD cells may partially overcome the loss of isoprenoid lipids by SREBP-mediated upregulation of enzymes required for isoprenoid biosynthesis. Furthermore, while inhibition of Rho/Rac/Rap prenylation promoted the release of IL-1β, specific inhibition of Rab prenylation by NE10790 had no effect in human peripheral blood mononuclear cells or human THP-1 monocytic cells. These studies demonstrate for the first time that mutations in mevalonate kinase can lead to a mild, temperature-induced defect in the prenylation of small GTPases, but that loss of prenylated Rab GTPases is not the cause of enhanced IL-1β release in MKD.

  4. Microstructures of beta-titanium orthodontic wires joined by infrared brazing.

    PubMed

    Iijima, Masahiro; Brantley, William A; Kawashima, Isao; Baba, Naoki; Alapati, Satish B; Yuasa, Toshihiro; Ohno, Hiroki; Mizoguchi, Itaru

    2006-10-01

    The microstructures and interdiffusion in brazed beta-titanium orthodontic wires were investigated by scanning electron microscopy and electron probe microanalysis, respectively. Beta-titanium wire (Ti-11Mo-6Zr-4Sn) with cross-section dimensions of 0.032 in. x 0.032 in., titanium-based braze alloy (Ti-30Ni-20Cu), and silver-based braze alloy (Ag-22Cu-17Zn-5Sn) were selected for the study. Brazing was performed using infrared radiation (RS-1) under an argon atmosphere. Specimens were etched with two solutions (2.5% HF + 2.5% HNO(3) + 95% H(2)O; 25% HN(4)OH + 30% H(2)O(2) + 45%H(2)O). It was found that the silver-based braze alloy has a eutectic structure. In the diffusion layer between the beta-titanium wire and this silver-based braze alloy, Cu and Ti were enriched on the wire side, and Sn and Ti were enriched on the braze alloy side. The titanium-based braze alloy has a dendritic structure. Beta-titanium wire specimens brazed with the titanium-based braze alloy had a thicker intermediate area compared to the silver alloy; Ti in the diffusion layer had an irregular concentration gradient, and the braze alloy side had higher Ti concentration. The original microstructure of the beta-titanium wire was not altered with the use of either braze alloy. Infrared brazing of beta-titanium orthodontic wire is acceptable for clinical use, since the wire microstructure did not deteriorate with either the titanium-based or silver-based braze alloy. The differing microstructures of the joint regions for the two braze alloys suggest that the joint strengths may also differ.

  5. Graphite-to-304SS Braze Joining by Active Metal-Brazing Technique: Improvement of Mechanical Properties

    NASA Astrophysics Data System (ADS)

    Ray, Ajoy K.; Kar, Abhijit; Kori, S. A.; Pathak, L. C.; Sonnad, A. N.

    2013-01-01

    In the present investigation, an attempt has been made to improve the mechanical strength of graphite-stainless steel-brazed joint. Due to high capillary action, the liquid filler alloy usually tends to percolate into the pores of graphite causing severe stress in the graphite near the joint interface resulting in poor joint strength of 10-15 MPa. In the present investigation, a thin coating of SiC was applied on graphite before the joining process to avoid the penetration of liquid filler alloy into the pores of the graphite. Active filler alloy Ag-Cu-Ti was used to braze the substrates. The brazing was carried out at 850, 900, 950, and 1000 °C. The characterization of the interfaces of the brazed joints was carried out using scanning electron microscopy attached with energy dispersive spectroscopy and x-ray diffraction analysis. From the correlation between the microstructural and mechanical properties, shear strength of approximately 35 MPa for graphite-304SS-brazed joint produced at 900 °C was demonstrated. After the shear tests, the fracture surfaces were analyzed by SEM-EDS.

  6. Radioisotope tracer studies in the NASA Skylab ethothermic brazing experiment M-552

    NASA Technical Reports Server (NTRS)

    Braski, D. N.; Adair, H. L.; Kobisk, E. H.

    1974-01-01

    The first use of radioisotope tracer for mapping flow patterns during brazing of metal components in a space environment (near-zero gravity) proved successful. A nickel ferrule was brazed to a nickel tube with Lithobraze BT (71.8% Ag, 28% Cu, 0.2% Li) which contained a trace amount of radioactive Ag-110. Mapping of the flow of the braze alloy in the annulus formed between the tube and the concentric ferrule was determined by counting the radiation intensity as a function of position in the braze joint. Significant information concerning the thermal history of the braze was determined.

  7. Brazing Inconel 625 Using Two Ni/(Fe)-Based Amorphous Filler Foils

    NASA Astrophysics Data System (ADS)

    Chen, Wen-Shiang; Shiue, Ren-Kae

    2012-07-01

    For MBF-51 filler, the brazed joint consists of interfacial grain boundary borides, coarse Nb6Ni16Si7, and Ni/Cr-rich matrix. In contrast, the VZ-2106 brazed joint is composed of interfacial Nb6Ni16Si7 precipitates as well as grain boundary borides, coarse Nb6Ni16Si7, and Ni/Cr/Fe-rich matrix. The maximum tensile strength of 443 MPa is obtained from the MBF-51 brazed specimen. The tensile strengths of VZ-2106 brazed joints are approximately 300 MPa. Both amorphous filler foils demonstrate potential in brazing IN-625 substrate.

  8. The Rab1 GTPase of Sciaenops ocellatus modulates intracellular bacterial infection.

    PubMed

    Hu, Yong-hua; Deng, Tian; Sun, Li

    2011-12-01

    The Rab family proteins belong to the Ras-like GTPase superfamily and play important roles in intracellular membrane trafficking. To date no studies on fish Rab have been documented, though rab-like sequences have been found in a number of teleosts. In this study, we identified and analyzed a Rab homologue, SoRab1, from red drum, Sciaenops ocellatus. The cDNA of SoRab1 contains a 5'- untranslated region (UTR) of 358 bp, an open reading frame (ORF) of 612 bp, and a 3'-UTR of 265 bp. The ORF encodes a putative protein of 203 residues, which shares 92-99% overall sequence identities with the Rab1 from fish, human, and mouse. SoRab1 possesses a typical Rab1 GTPase domain with the conserved G box motifs and the switch I and switch II regions. Recombinant SoRab1 purified from Escherichia coli exhibits apparent GTPase activity. Quantitative real time RT-PCR analysis showed that SoRab1 expression was detected in a number of tissues, with the lowest expression found in blood and highest expression found in muscle. Bacterial and lipopolysaccharide challenges significantly upregulated SoRab1 expression in liver, kidney, and spleen in time-dependent manners. Transient overexpression of SoRab1 in primary hepatocytes reduced intracellular bacterial infection, whereas interference with SoRab1 expression by RNAi enhanced intracellular bacterial invasion. These results provide the first indication that a fish Rab1 GTPase, SoRab1, regulates intracellular bacterial infection and thus is likely to play a role in bacteria-induced host immune defense.

  9. A review of oxide, silicon nitride, and silicon carbide brazing

    SciTech Connect

    Santella, M.L.; Moorhead, A.J.

    1987-01-01

    There is growing interest in using ceramics for structural applications, many of which require the fabrication of components with complicated shapes. Normal ceramic processing methods restrict the shapes into which these materials can be produced, but ceramic joining technology can be used to overcome many of these limitations, and also offers the possibility for improving the reliability of ceramic components. One method of joining ceramics is by brazing. The metallic alloys used for bonding must wet and adhere to the ceramic surfaces without excessive reaction. Alumina, partially stabilized zirconia, and silicon nitride have high ionic character to their chemical bonds and are difficult to wet. Alloys for brazing these materials must be formulated to overcome this problem. Silicon carbide, which has some metallic characteristics, reacts excessively with many alloys, and forms joints of low mechanical strength. The brazing characteristics of these three types of ceramics, and residual stresses in ceramic-to-metal joints are briefly discussed.

  10. Development of the copper-tin diffusion-brazing process

    SciTech Connect

    Sangha, S.P.S.; Jacobson, D.M.; Peacock, A.T.

    1998-10-01

    The copper-tin diffusion-brazing process has been studied with the objective of applying it to the joining of plasma-facing beryllium tiles to copper-based heat sinks in a nuclear-fusion reactor. The process is silver-free -- an essential requirement for this application -- and can be carried out at temperatures below 700 C (1,292 F). This approach produces thin joints of essentially pure copper of high thermal conductance with the requisite strength. Satisfactory conditions for achieving robust joints under the constraints demanded by the nuclear-fusion application have been established. The roles of the process parameters -- thickness of the filler metal tin, the compressive loading applied to the components during the brazing cycle and the brazing temperature -- have been assessed.

  11. Development of Al sbnd Ge base rheocast brazing alloys

    NASA Astrophysics Data System (ADS)

    Saint-Antonin, F.; Suéry, M.; Meneses, P.; Le Marois, G.; Moret, F.

    1996-10-01

    This study concerns the development of an aluminium—germanium rheocast brazing alloy able to join replaceable sacrificial protections onto permanently cooled substrates in the ITER divertor. The rheocast alloy must be solid at the in-service temperature (up to 350°C) and semi-solid, with a viscosity close to that of grease, in a wide range of temperature above: this last property will allow the in-situ replacement of the sacrificial components even on vertical surfaces. This paper describes the fabrication route of the rheocast and reports the mechanical characterisation both up to 400°C and in the semi-solid region. Moreover, the brazing process has been optimized to obtain sound joints. The surface, mechanical resistance and thermal stability of the Al sbnd Ge/Cu joint are presented. Successive brazing tests have been performed to show the feasibility to sacrificial parts removal and replacement.

  12. Rab11 Regulates the Mast Cell Exocytic Response.

    PubMed

    Wilson, Joshua D; Shelby, Sarah A; Holowka, David; Baird, Barbara

    2016-09-01

    Stimulated exocytic events provide a means for physiological communication and are a hallmark of the mast cell-mediated allergic response. In mast cells these processes are triggered by antigen crosslinking of IgE bound to its high-affinity receptor, FcϵRI, on the cell surface. Here we use the endosomal v-SNARE VAMP8, and the lysosomal hydrolase β-hexosaminidase (β-Hex), each C-terminally fused to super-ecliptic pHluorin, to monitor stimulated exocytosis. Using these pHluorin-tagged constructs, we monitor stimulated exocytosis by fluorimetry and visualize individual exocytic events with total internal reflection (TIRF) microscopy. Similar to constitutive recycling endosome (RE) trafficking, we find that stimulated RE exocytosis, monitored by VAMP8, is attenuated by expression of dominant negative (S25N) Rab11. Stimulated β-Hex exocytosis is also reduced in the presence of S25N Rab11, suggesting that expression of this mutant broadly impacts exocytosis. Interestingly, pretreatment with inhibitors of actin polymerization, cytochalasin D or latrunculin A, substantially restores both RE and lysosome exocytosis in cells expressing S25N Rab11. Conversely, stabilizing F-actin with jasplakinolide inhibits antigen-stimulated exocytosis but is not additive with S25N Rab11-mediated inhibition, suggesting that these reagents inhibit related processes. Together, our results suggest that Rab11 participates in the regulation necessary for depolymerization of the actin cytoskeleton during stimulated exocytosis in mast cells.

  13. High Temperature Strength of YSZ Joints Brazed with Palladium Silver Copper Oxide Filler Metals

    SciTech Connect

    Darsell, Jens T.; Weil, K. Scott

    2010-06-09

    The Ag-CuOx system is being investigated as potential filler metals for use in air brazing high-temperature electrochemical devices such as solid oxide fuel cells and gas concentrators. The current study examines the effects of palladium addition on the high temperature joint strength of specimens prepared from yttria stabilized zirconia (YSZ) bars brazed with the binary Ag-CuOx, and 15Pd-Ag-CuO. It was found that while the binary Ag-CuOx system exhibits stronger room temperature strength than the 15Pd system the strength is reduced to values equivalent of the 15Pd system at 800°C. The 15Pd system exhibits a lower ambient temperature strength that is retained at 800°C. In both systems the failure mechanism at high temperature appears to be peeling of the noble metal component from the oxide phases and tearing through the noble metal phase whereas sufficient adhesion is retained at lower temperatures to cause fracture of the YSZ substrate.

  14. Infrared Brazing Fe3Al Using Ag-Based Filler Metals

    NASA Astrophysics Data System (ADS)

    Shiue, Ren-Kae; Li, Yao; Wu, Shyi-Kaan; Wu, Ling-Mei

    2010-11-01

    The microstructural evolution and bonding shear strength of infrared brazed Fe3Al using Ag and BAg-8 (72Ag-28Cu in wt pct) braze alloys have been studied. The Ag-rich phase alloyed with Al dominates the entire Ag brazed joints, and the shear strength is independent of the brazing time. The BAg-8 brazed joint contains Ag-Cu eutectic for all brazing conditions, and its shear strength increases slightly with increasing brazing time. The highest shear strength of 181 MPa is acquired from the joint infrared brazed at 1073 K (800 °C) for 600 seconds. A thin layer of Fe3Al is identified at the interface between the brazed zone and the substrate for both braze alloys. An Al depletion zone in the Fe3Al substrate next to the interfacial Fe3Al is identified as the α-Fe phase. The dissolution of Al from the Fe3Al substrate into the molten braze causes the formation of α-Fe in the Fe3Al substrate.

  15. Interfacial Microstructure Evolution and Shear Strength of Titanium Sandwich Structures Fabricated by Brazing

    NASA Astrophysics Data System (ADS)

    Wang, Wentao; Fan, Minyu; Li, Jinlong; Tao, Jie

    2016-03-01

    The corrugated sandwich structure, consisting of a CP Ti (commercially pure titanium) core between two Ti-6Al-4V face sheets, was brazed using pasty Ti-37.5Zr-15Cu-10Ni as filler alloy, at the temperature of 870°C for 5, 10, 20, and 30 min. The effect of brazing time on the microstructure and elemental distribution of the brazed joints was examined by means of SEM, EDS, and XRD analyses. It was found that various intermetallic phases were formed in the brazed joints, following a brazing time of 5 min, and their contents were decreased by the increment of brazing time, while prolonged brazing time resulted in a fine, acicular Widmanstätten microstructure throughout the entire joint. In addition, shear testing was performed in the brazed corrugated specimens in order to indirectly assess the quality of the joints. The debonding between CP Ti and Ti-6Al-4V was observed in the specimen brazed for 5 min and the fracture of the CP Ti corrugated core occurred after 30 min of brazing time. Additionally, when brazed for 10 min or 20 min, brittle intermetallic compounds in the joints and the grain growth of the base metal were controllable. Therefore, the sandwich structures failed without debonding in the joints or fracture within the base metal, demonstrating a good combination of strength and ductility.

  16. Genetic Analysis of Yeast Yip1p Function Reveals a Requirement for Golgi-Localized Rab Proteins and Rab-Guanine Nucleotide Dissociation Inhibitor

    PubMed Central

    Chen, Catherine Z.; Calero, Monica; DeRegis, Carol J.; Heidtman, Matthew; Barlowe, Charles; Collins, Ruth N.

    2004-01-01

    Yip1p is the first identified Rab-interacting membrane protein and the founder member of the YIP1 family, with both orthologs and paralogs found in all eukaryotic genomes. The exact role of Yip1p is unclear; YIP1 is an essential gene and defective alleles severely disrupt membrane transport and inhibit ER vesicle budding. Yip1p has the ability to physically interact with Rab proteins and the nature of this interaction has led to suggestions that Yip1p may function in the process by which Rab proteins translocate between cytosol and membranes. In this study we have investigated the physiological requirements for Yip1p action. Yip1p function requires Rab-GDI and Rab proteins, and several mutations that abrogate Yip1p function lack Rab-interacting capability. We have previously shown that Yip1p in detergent extracts has the capability to physically interact with Rab proteins in a promiscuous manner; however, a genetic analysis that covers every yeast Rab reveals that the Rab requirement in vivo is exclusively confined to a subset of Rab proteins that are localized to the Golgi apparatus. PMID:15611160

  17. Genetic analysis of yeast Yip1p function reveals a requirement for Golgi-localized rab proteins and rab-Guanine nucleotide dissociation inhibitor.

    PubMed

    Chen, Catherine Z; Calero, Monica; DeRegis, Carol J; Heidtman, Matthew; Barlowe, Charles; Collins, Ruth N

    2004-12-01

    Yip1p is the first identified Rab-interacting membrane protein and the founder member of the YIP1 family, with both orthologs and paralogs found in all eukaryotic genomes. The exact role of Yip1p is unclear; YIP1 is an essential gene and defective alleles severely disrupt membrane transport and inhibit ER vesicle budding. Yip1p has the ability to physically interact with Rab proteins and the nature of this interaction has led to suggestions that Yip1p may function in the process by which Rab proteins translocate between cytosol and membranes. In this study we have investigated the physiological requirements for Yip1p action. Yip1p function requires Rab-GDI and Rab proteins, and several mutations that abrogate Yip1p function lack Rab-interacting capability. We have previously shown that Yip1p in detergent extracts has the capability to physically interact with Rab proteins in a promiscuous manner; however, a genetic analysis that covers every yeast Rab reveals that the Rab requirement in vivo is exclusively confined to a subset of Rab proteins that are localized to the Golgi apparatus.

  18. Overexpression of Rab22a hampers the transport between endosomes and the Golgi apparatus

    SciTech Connect

    Mesa, Rosana; Magadan, Javier; Barbieri, Alejandro; Lopez, Cecilia; Stahl, Philip D.; Mayorga, Luis S. . E-mail: lmayorga@fcm.uncu.edu.ar

    2005-04-01

    The transport and sorting of soluble and membrane-associated macromolecules arriving at endosomal compartments require a complex set of Rab proteins. Rab22a has been localized to the endocytic compartment; however, very little is known about the function of Rab22a and inconsistent results have been reported in studies performed in different cell lines. To characterize the function of Rab22a in endocytic transport, the wild-type protein (Rab22a WT), a hydrolysis-deficient mutant (Rab22a Q64L), and a mutant with reduced affinity for GTP (Rab22a S19N) were expressed in CHO cells. None of the three Rab22a constructs affected the transport of rhodamine-dextran to lysosomes, the digestion of internalized proteins, or the lysosomal localization of cathepsin D. In contrast with the mild effect of Rab22a on the endosome-lysosome route, cells expressing Rab22a WT and Rab22a Q64L presented a strong delay in the retrograde transport of cholera toxin from endosomes to the Golgi apparatus. Moreover, these cells accumulated the cation independent mannose 6-phosphate receptor in endosomes. These observations indicate that Rab22a can affect the trafficking from endosomes to the Golgi apparatus probably by promoting fusion among endosomes and impairing the proper segregation of membrane domains required for targeting to the trans-Golgi network (TGN)

  19. Rab22 controls NGF signaling and neurite outgrowth in PC12 cells.

    PubMed

    Wang, Liang; Liang, Zhimin; Li, Guangpu

    2011-10-01

    Rab22 is a small GTPase that is localized on early endosomes and regulates early endosomal sorting. This study reports that Rab22 promotes nerve growth factor (NGF) signaling-dependent neurite outgrowth and gene expression in PC12 cells by sorting NGF and the activated/phosphorylated receptor (pTrkA) into signaling endosomes to sustain signal transduction in the cell. NGF binding induces the endocytosis of pTrkA into Rab22-containing endosomes. Knockdown of Rab22 via small hairpin RNA (shRNA) blocks NGF-induced pTrkA endocytosis into the endosomes and gene expression (VGF) and neurite outgrowth. Overexpression of human Rab22 can rescue the inhibitory effects of the Rab22 shRNA, suggesting a specific Rab22 function in NGF signal transduction, rather than off-target effects. Furthermore, the Rab22 effector, Rabex-5, is necessary for NGF-induced neurite outgrowth and gene expression, as evidenced by the inhibitory effect of shRNA-mediated knockdown of Rabex-5. Disruption of the Rab22-Rabex-5 interaction via overexpression of the Rab22-binding domain of Rabex-5 in the cell also blocks NGF-induced neurite outgrowth, suggesting a critical role of Rab22-Rabex-5 interaction in the biogenesis of NGF-signaling endosomes to sustain the signal for neurite outgrowth. These data provide the first evidence for an early endosomal Rab GTPase as a positive regulator of NGF signal transduction and cell differentiation.

  20. Joining a Ni-based creep-resistant (ODS) alloy by brazing

    NASA Astrophysics Data System (ADS)

    Bucklow, I. A.

    Joints are produced in a fine-grained Ni oxide-dispersion-strengthened (ODS) alloy by means of brazing alloys with and without boron, and the resulting joints are studied mechanographically. The brazing alloys employed are either sputtered coatings or foils, and brazing is conducted in a vacuum for approximately two hours. The resulting joints are examined during the process by mass spectrometry and afterwards by means of metallographic observation following etching in glyceregia. Premature and uncontrolled recrystallization of the parent metal is noted in the samples brazed with alloys obtaining boron. The 2-micron braze coating used minimize second-phase formation and dispersoid agglomerations, and the 25-micron brazing foils lead to high porosity and dispersoid aggregation due to excessive melting. Recrystallization of the parent metal near brazing zones is concluded to be undesirable although is does not necessarily influence the quality of the joint.

  1. Brazing diamond grits onto a steel substrate using copper alloys as the filler metals

    NASA Astrophysics Data System (ADS)

    Chen, S.-M.; Lin, S.-T.

    1996-12-01

    Surface-set diamond tools were fabricated by an active metal brazing process, using bronze (Cu-8.9Sn) powder and 316L stainless steel powder mixed to various ratios as the braze filler metals. The diamond grits were brazed onto a steel substrate at 1050 °C for 30 min in a dry hydrogen atmosphere. After brazing practice, an intermediate layer rich in chromium formed between the braze filler metal and diamond. A braze filler metal composed of 70 wt % bronze powder and 30 wt % stainless steel powder was found to be optimum in that the diamond grits were strongly impregnated in the filler metal by both mechanical and chemical types of holding. The diamond tools thus fabricated performed better than conventional nickel-plated diamond tools. In service, the braze filler metal wore at almost the same rate as the diamond grits, and no pullout of diamond grits or peeling of the filler metal layer took place.

  2. Computational simulations and experimental validation of a furnace brazing process

    SciTech Connect

    Hosking, F.M.; Gianoulakis, S.E.; Malizia, L.A.

    1998-12-31

    Modeling of a furnace brazing process is described. The computational tools predict the thermal response of loaded hardware in a hydrogen brazing furnace to programmed furnace profiles. Experiments were conducted to validate the model and resolve computational uncertainties. Critical boundary conditions that affect materials and processing response to the furnace environment were determined. {open_quotes}Global{close_quotes} and local issues (i.e., at the furnace/hardware and joint levels, respectively) are discussed. The ability to accurately simulate and control furnace conditions is examined.

  3. Development of a Cu-Sn based brazing system with a low brazing and a high remelting temperature

    NASA Astrophysics Data System (ADS)

    Schmieding, M.; Holländer, U.; Möhwald, K.

    2017-03-01

    Objective of the project presented is the development of a joining process for hot working steel components at low brazing temperatures leading to a bond with a much higher remelting temperature. This basically is achieved by the use of a Cu-Sn melt spinning foil combined with a pure Cu foil. During brazing, the Sn content of the foil is decreased by diffusion of Sn into the additional Cu resulting in a homogenious joint with a increased remelting temperature of the filler metal. Within this project specimens were brazed and diffusion annealed in a vacuum furnace at 850 °C varying the processing times (0 – 10 h). The samples prepared were studied metallographically and diffusion profiles of Sn were recorded using EDX line scans. The results are discussed in view of further investigations and envisaged applications.

  4. Effect of the Secretory Small GTPase Rab27B on Breast Cancer Growth, Invasion, and Metastasis

    PubMed Central

    Hendrix, An; Maynard, Dawn; Pauwels, Patrick; Braems, Geert; Denys, Hannelore; Van den Broecke, Rudy; Lambert, Jo; Van Belle, Simon; Cocquyt, Veronique; Gespach, Christian; Bracke, Marc; Seabra, Miguel C.; Gahl, William A.

    2010-01-01

    Background Secretory GTPases like Rab27B control vesicle exocytosis and deliver critical proinvasive growth regulators into the tumor microenvironment. The expression and role of Rab27B in breast cancer were unknown. Methods Expression of green fluorescent protein (GFP) fused with wild-type Rab3D, Rab27A, or Rab27B, or Rab27B point mutants defective in GTP/GDP binding or geranylgeranylation, or transient silencing RNA to the same proteins was used to study Rab27B in estrogen receptor (ER)–positive human breast cancer cell lines (MCF-7, T47D, and ZR75.1). Cell cycle progression was evaluated by flow cytometry, western blotting, and measurement of cell proliferation rates, and invasion was assessed using Matrigel and native type I collagen substrates. Orthotopic tumor growth, local invasion, and metastasis were analyzed in mouse xenograft models. Mass spectrometry identified proinvasive growth regulators that were secreted in the presence of Rab27B. Rab27B protein levels were evaluated by immunohistochemistry in 59 clinical breast cancer specimens, and Rab3D, Rab27A, and Rab27B mRNA levels were analyzed by quantitative real-time polymerase chain reaction in 20 specimens. Statistical tests were two-sided. Results Increased expression of Rab27B promoted G1 to S phase cell cycle transition, proliferation and invasiveness of cells in culture, and invasive tumor growth and hemorrhagic ascites production in a xenograft mouse model (n = 10; at 10 weeks, survival of MCF-7 GFP- vs GFP-Rab27B–injected mice was 100% vs 62.5%, hazard ratio = 0.26, 95% confidence interval = 0.08 to 0.88, P = .03). Mass spectrometric analysis of purified Rab27B-secretory vesicles identified heat-shock protein 90α as key proinvasive growth regulator. Heat-shock protein 90α secretion was Rab27B-dependent and was required for matrix metalloproteinase-2 activation. All Rab27B-mediated functional responses were GTP- and geranylgeranyl-dependent. Presence of endogenous Rab27B mRNA and protein, but

  5. Characterization of RAB-like4, the first identified RAB-like protein from Trypanosoma cruzi with GTPase activity.

    PubMed

    Ramos, Fabiane Pereira; Araripe, Júlia Rolão; Urményi, Turán Péter; Silva, Rosane; Cunha e Silva, Narcisa Leal; Leite Fontes, Carlos Frederico; da Silveira, José Franco; Rondinelli, Edson

    2005-08-05

    RAB proteins, which belong to the RAS superfamily, regulate exocytic and endocytic pathways of eukaryotic cells, controlling vesicle docking and fusion. Few RAB proteins have been identified in parasites. Molecular markers for cellular compartments are important to studies concerning about the protein traffic in Trypanosoma cruzi, the causal agent of Chagas disease. In this work, we describe the characterization of TcRABL4, the first RAB-like gene identified in T. cruzi (GenBank Accession No.: ), present as a single-copy gene. TcRABL4 contains all five consensus RAB motifs but lacks cysteine residues at the C terminus, which are essential to isoprenylation, an absolute prerequisite for membrane association of these proteins. TcRABL4 is a functional GTPase that is able to bind and hydrolyze GTP, and its gene is transcribed as a single 1.2 kb mRNA in epimastigotes. TcRABL4 appears to be differentially regulated in the three cell forms of the parasite, and the protein is not associated to membranes, unlike other RAB proteins. It is possible that TcRABL4 may be a member of a novel family of small GTPases.

  6. The Rab11-FIP1/RCP gene codes for multiple protein transcripts related to the plasma membrane recycling system.

    PubMed

    Jin, Min; Goldenring, James R

    2006-06-01

    Rab11a is a member of the Rab11 small GTPase family, and plays an important role in plasma membrane recycling. Rab11-Family Interacting Protein 1 (Rab11-FIP1) binds to Rab11 through a carboxyl-terminal amphipathic alpha helix. We have identified eight alternatively spliced Rab11-FIP1 gene transcripts from human chromosome 8. Among them, Rab11-FIP1A-D have carboxyl terminal Rab11 binding domains, while Rab11-FIP1E-H do not contain the Rab11 binding domain. While Rab11-FIP1B and F gene transcripts are ubiquitous, other Rab11-FIP1 transcripts demonstrate more limited patterns of expression in human tissue cDNAs. EGFP-Rab11-FIP1A-D proteins over-expressed in HeLa cells targeted to Rab11a-containing membranes, while EGFP-Rab11-FIP1E/F and H proteins did not localize with recycling system membranes. However, transferrin trafficking was not significantly altered in HeLa cells over-expressing expressing any of the EGFP-Rab11-FIP1 proteins. Rabbit polyclonal antibodies specific for Rab11-FIP1B and Rab11-FIP1C/RCP demonstrated that Rab11-FIP1B and Rab11-FIP1C/RCP are expressed endogenously. Strikingly, endogenous staining for Rab11-FIP1C/RCP only partially co-localized with EGFP-Rab11-FIP1A, EGFP-Rab11-FIP1B, and EGFP-Rab11a in the perinuclear region, indicating that Rab11-FIP1C/RCP resides in a differentiable subcellular compartment within the plasma membrane recycling system compared with Rab11-FIP1A and Rab11-FIP1B. These data suggest that Rab11-FIP1 proteins may play coordinated roles in regulating plasma membrane recycling with regional specificity within the Rab11a-containing recycling system.

  7. Role for Rab10 in Methamphetamine-Induced Behavior.

    PubMed

    Vanderwerf, Scott M; Buck, David C; Wilmarth, Phillip A; Sears, Leila M; David, Larry L; Morton, David B; Neve, Kim A

    2015-01-01

    Lipid rafts are specialized, cholesterol-rich membrane compartments that help to organize transmembrane signaling by restricting or promoting interactions with subsets of the cellular proteome. The hypothesis driving this study was that identifying proteins whose relative abundance in rafts is altered by the abused psychostimulant methamphetamine would contribute to fully describing the pathways involved in acute and chronic effects of the drug. Using a detergent-free method for preparing rafts from rat brain striatal membranes, we identified density gradient fractions enriched in the raft protein flotillin but deficient in calnexin and the transferrin receptor, markers of non-raft membranes. Dopamine D1- and D2-like receptor binding activity was highly enriched in the raft fractions, but pretreating rats with methamphetamine (2 mg/kg) once or repeatedly for 11 days did not alter the distribution of the receptors. LC-MS analysis of the protein composition of raft fractions from rats treated once with methamphetamine or saline identified methamphetamine-induced changes in the relative abundance of 23 raft proteins, including the monomeric GTP-binding protein Rab10, whose abundance in rafts was decreased 2.1-fold by acute methamphetamine treatment. Decreased raft localization was associated with a selective decrease in the abundance of Rab10 in a membrane fraction that includes synaptic vesicles and endosomes. Inhibiting Rab10 activity by pan-neuronal expression of a dominant-negative Rab10 mutant in Drosophila melanogaster decreased methamphetamine-induced activity and mortality and decreased caffeine-stimulated activity but not mortality, whereas inhibiting Rab10 activity selectively in cholinergic neurons had no effect. These results suggest that activation and redistribution of Rab10 is critical for some of the behavioral effects of psychostimulants.

  8. Role for Rab10 in Methamphetamine-Induced Behavior

    PubMed Central

    Vanderwerf, Scott M.; Buck, David C.; Wilmarth, Phillip A.; Sears, Leila M.; David, Larry L.; Morton, David B.; Neve, Kim A.

    2015-01-01

    Lipid rafts are specialized, cholesterol-rich membrane compartments that help to organize transmembrane signaling by restricting or promoting interactions with subsets of the cellular proteome. The hypothesis driving this study was that identifying proteins whose relative abundance in rafts is altered by the abused psychostimulant methamphetamine would contribute to fully describing the pathways involved in acute and chronic effects of the drug. Using a detergent-free method for preparing rafts from rat brain striatal membranes, we identified density gradient fractions enriched in the raft protein flotillin but deficient in calnexin and the transferrin receptor, markers of non-raft membranes. Dopamine D1- and D2-like receptor binding activity was highly enriched in the raft fractions, but pretreating rats with methamphetamine (2 mg/kg) once or repeatedly for 11 days did not alter the distribution of the receptors. LC-MS analysis of the protein composition of raft fractions from rats treated once with methamphetamine or saline identified methamphetamine-induced changes in the relative abundance of 23 raft proteins, including the monomeric GTP-binding protein Rab10, whose abundance in rafts was decreased 2.1-fold by acute methamphetamine treatment. Decreased raft localization was associated with a selective decrease in the abundance of Rab10 in a membrane fraction that includes synaptic vesicles and endosomes. Inhibiting Rab10 activity by pan-neuronal expression of a dominant-negative Rab10 mutant in Drosophila melanogaster decreased methamphetamine-induced activity and mortality and decreased caffeine-stimulated activity but not mortality, whereas inhibiting Rab10 activity selectively in cholinergic neurons had no effect. These results suggest that activation and redistribution of Rab10 is critical for some of the behavioral effects of psychostimulants. PMID:26291453

  9. Rice Rab11 is required for JA-mediated defense signaling

    SciTech Connect

    Hong, Min Ji; Lee, Yun mi; Son, Young Sim; Im, Chak Han; Yi, Young Byung; Rim, Yeong Gil; Bahk, Jeong Dong; Heo, Jae Bok

    2013-05-17

    Highlights: •OsRab11 interacts with OsOPR8. •OsOPR8 is localized in the cytosol and peroxisome. •OsRab11 enhances the NADPH consumption by OsOPR8. •Transgenic Arabidopsis overexpressing OsRab11 represents a pathogen-resistant phenotype. -- Abstract: Rab proteins play an essential role in regulating vesicular transport in eukaryotic cells. Previously, we characterized OsRab11, which in concert with OsGAP1 and OsGDI3 regulates vesicular trafficking from the trans-Golgi network (TGN) to the plasma membrane or vacuole. To further elucidate the physiological function of OsRab11 in plants, we performed yeast two-hybrid screens using OsRab11 as bait. OsOPR8 was isolated and shown to interact with OsRab11. A co-immunoprecipitation assay confirmed this interaction. The green fluorescent protein-OsOPR8 fusion product was targeted to the cytoplasm and peroxisomes of protoplasts from Arabidopsis thaliana. OsOPR8 exhibited NADPH-dependent reduction activity when 2-cyclohexen-1-one (CyHE) and 12-oxo-phytodienoic acid (OPDA) were supplied as possible substrates. Interestingly, NADPH oxidation by OsOPR8 was increased when wild-type OsRab11 or the constitutively active form of OsRab11 (Q78L) were included in the reaction mix, but not when the dominant negative form of OsRab11 (S28N) was included. OsRab11 was expressed broadly in plants and both OsRab11 and OsOPR8 were induced by jasmonic acid (JA) and elicitor treatments. Overexpressed OsRab11 transgenic plants showed resistance to pathogens through induced expression of JA-responsive genes. In conclusion, OsRab11 may be required for JA-mediated defense signaling by activating the reducing activity of OsOPR8.

  10. Trypanosoma cruzi: TcRAB7 protein is localized at the Golgi apparatus in epimastigotes.

    PubMed

    Araripe, Júlia R; Cunha e Silva, Narcisa L; Leal, Simone T; de Souza, Wanderley; Rondinelli, Edson

    2004-08-20

    In mammalian cells, the Rab7 protein is a key element of late endocytic membrane traffic. Several results suggest that it is involved in the transport from early to late endosome or from late endosome to lysosome. We have previously characterized a Rab7 gene homologue (TcRAB7) in Trypanosoma cruzi. Now, using an affinity-purified antibody specific to TcRAB7 protein we have determined that it is localized at the Golgi apparatus of the parasite. Our results indicate that the T. cruzi Rab7 homologue may function in a different route than its counterparts in mammalian cells.

  11. Loss of RAB-3/A in Caenorhabditis elegans and the mouse affects behavioral response to ethanol.

    PubMed

    Kapfhamer, D; Bettinger, J C; Davies, A G; Eastman, C L; Smail, E A; Heberlein, U; McIntire, S L

    2008-08-01

    The mechanisms by which ethanol induces changes in behavior are not well understood. Here, we show that Caenorhabditis elegans loss-of-function mutations in the synaptic vesicle-associated RAB-3 protein and its guanosine triphosphate exchange factor AEX-3 confer resistance to the acute locomotor effects of ethanol. Similarly, mice lacking one or both copies of Rab3A are resistant to the ataxic and sedative effects of ethanol, and Rab3A haploinsufficiency increases voluntary ethanol consumption. These data suggest a conserved role of RAB-3-/RAB3A-regulated neurotransmitter release in ethanol-related behaviors.

  12. Is Rab25 a tumor promoter or suppressor--context dependency on RCP status?

    PubMed

    Tang, Bor Luen

    2010-08-01

    Conflicting reports in the literature suggest that Rab25 could either be a context dependent promoter or suppressor of tumorigenesis. We hypothesized that whether Rab25 acts as a promoter or suppressor in tumor progression depends on the expression status of its effector, the Rab coupling protein (RCP). An elevated expression of RCP resulting from genomic amplification may enhance Rab25's tumor progression activity. Elevation of Rab25 alone may sequester endogenous RCP, and attenuates its activating effect on other oncogenic products, such as mutant Ras.

  13. The Interaction of JRAB/MICAL-L2 with Rab8 and Rab13 Coordinates the Assembly of Tight Junctions and Adherens Junctions

    PubMed Central

    Yamamura, Rie; Nishimura, Noriyuki; Nakatsuji, Hiroyoshi; Arase, Seiji

    2008-01-01

    The assembly of tight junctions (TJs) and adherens junctions (AJs) is regulated by the transport of integral TJ and AJ proteins to and/or from the plasma membrane (PM) and it is tightly coordinated in epithelial cells. We previously reported that Rab13 and a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) mediated the endocytic recycling of an integral TJ protein occludin and the formation of functional TJs. Here, we investigated the role of Rab13 and JRAB/MICAL-L2 in the transport of other integral TJ and AJ proteins claudin-1 and E-cadherin to the PM by using a Ca2+-switch model. Although knockdown of Rab13 specifically suppressed claudin-1 and occludin but not E-cadherin transport, knockdown of JRAB/MICAL-L2 and expression of its Rab13-binding domain (JRAB/MICAL-L2-C) inhibited claudin-1, occludin, and E-cadherin transport. We then identified Rab8 as another JRAB/MICAL-L2-C-binding protein. Knockdown of Rab8 inhibited the Rab13-independent transport of E-cadherin to the PM. Rab8 and Rab13 competed with each other for the binding to JRAB/MICAL-L2 and functionally associated with JRAB/MICAL-L2 at the perinuclear recycling/storage compartments and PM, respectively. These results suggest that the interaction of JRAB/MICAL-L2 with Rab8 and Rab13 coordinates the assembly of AJs and TJs. PMID:18094055

  14. More About Brazing Or Welding NiAl Without Filler

    NASA Technical Reports Server (NTRS)

    Moore, Thomas J.; Kalinowski, Joseph M.

    1996-01-01

    Two reports present additional information about two processes for joining, brazing, or welding workpieces made of nickel aluminide alloys, without use of filler metal. Joining processes involve uniform heating in vacuum-controlled furnace. Eliminates internal thermal gradients in workpieces joined and greatly reduces tendency toward cracking.

  15. Braze alloy holds bonding strength over wide temperature range

    NASA Technical Reports Server (NTRS)

    1966-01-01

    Copper-based quaternary alloys of the solid solution type is used for vacuum furnace brazing of large stainless steel components at a maximum temperature of 1975 deg F. The alloy has high bonding strength and good ductility over a temperature range extending from the cryogenic region to approximately 800 deg F.

  16. On the metallurgy of active brazing of silicon nitride

    SciTech Connect

    Paulasto, M.; Kivilahti, J.K.; Loo, F.J.J. van

    1996-10-01

    Activation mechanism and interfacial reactions in brazing of Si{sub 3}N{sub 4} with different AgCuTi filler alloys has been investigated thermodynamically and experimentally. During brazing the AgCuTi filler alloy is divided into two liquids L1 and L2, where the (Cu,Ti)-rich liquid L2 gathers at the Si{sub 3}N{sub 4} interface and reacts with it. The concentration of Ti in L2 is high, but due to the strong attractive interaction between Cu and Ti, the activity of Ti in L2 is considerably low. In reaction between Si{sub 3}N{sub 4} and Cu-containing brazes at Ti-Cu-Si-N compound is typically formed at the interface. In absence of Cu, in joints brazed with AgTi based filler alloys, extremely Ti-rich solution reacts with Si{sub 3}N{sub 4} and as reaction products TiN and Ti{sub 5}Si{sub 3}[N] are formed.

  17. Structural Performance of Inconel 625 Superalloy Brazed Joints

    NASA Astrophysics Data System (ADS)

    Chen, Jianqiang; Demers, Vincent; Cadotte, Eve-Line; Turner, Daniel; Bocher, Philippe

    2016-12-01

    The purpose of this work was to investigate tensile and fatigue behaviors of Inconel 625 superalloy brazed joints after transient liquid-phase bonding process. Brazing was performed in a vacuum furnace using a nickel-based filler metal in a form of paste to join wrought Inconel 625 plates. Mechanical tests were carried out on single-lap joints under various lap distance-to-thickness ratios. The fatigue crack initiation and crack growth modes were examined via metallographic analysis, and the effect of local stress on fatigue life was assessed by finite element simulations. The fatigue results show that fatigue strength and endurance limit increase with overlap distance, leading to a relatively large scatter of results. Fatigue cracks nucleated in the high-stressed region of the weld fillets from brittle eutectic phases or from internal brazing cavities. The present work proposes to rationalize the results by using the local stress at the brazing fillet. When using this local stress, all fatigue-obtained results find themselves on a single S-N curve, providing a design curve for any joint configuration in fatigue solicitation.

  18. Coating method enables low-temperature brazing of stainless steel

    NASA Technical Reports Server (NTRS)

    Seaman, F. D.

    1965-01-01

    Gold coated stainless steel tubes containing insulated electrical conductors are brazed at a low temperature to a copper coated stainless steel sealing block with a gold-copper eutectic. This produces an effective seal without using flux or damaging the electrical conductors.

  19. Structural Performance of Inconel 625 Superalloy Brazed Joints

    NASA Astrophysics Data System (ADS)

    Chen, Jianqiang; Demers, Vincent; Cadotte, Eve-Line; Turner, Daniel; Bocher, Philippe

    2017-02-01

    The purpose of this work was to investigate tensile and fatigue behaviors of Inconel 625 superalloy brazed joints after transient liquid-phase bonding process. Brazing was performed in a vacuum furnace using a nickel-based filler metal in a form of paste to join wrought Inconel 625 plates. Mechanical tests were carried out on single-lap joints under various lap distance-to-thickness ratios. The fatigue crack initiation and crack growth modes were examined via metallographic analysis, and the effect of local stress on fatigue life was assessed by finite element simulations. The fatigue results show that fatigue strength and endurance limit increase with overlap distance, leading to a relatively large scatter of results. Fatigue cracks nucleated in the high-stressed region of the weld fillets from brittle eutectic phases or from internal brazing cavities. The present work proposes to rationalize the results by using the local stress at the brazing fillet. When using this local stress, all fatigue-obtained results find themselves on a single S- N curve, providing a design curve for any joint configuration in fatigue solicitation.

  20. Amorphous Ti-Zr; Base Metglas brazing filler metals

    SciTech Connect

    Rabinkin, A.; Liebermann, H.; Pounds, S.; Taylor, T. )

    1991-01-01

    This paper is the first report on processing, properties and potential application of amorphous titanium/zirconium-base alloys produced in the form of a good quality continuous and ductile ribbon having up to 12.5 mm width. To date, the majority of titanium brazing is accomplished using cooper and aluminum-base brazing filler metals. The brazements produced with these filler metals have rather low ({approximately}300{degrees} C) service temperature, thus impeding progress in aircraft and other technologies and industries. The attempt to develop a generation of high temperature brazing filler metals was made in the late sixties-early seventies studies in detail were a large number of Ti-, Zr-Ti-Zr, Ti-V and Zr-V-Ti based alloys. The majority of these alloys has copper and nickel as melting temperature depressants. The presence of nickel and copper converts them into eutectic alloys having (Ti(Zr)) (Cu(Ni)), intermetallic phases as major structural constituents. This, in turn, results in high alloy brittleness and poor, if any, processability by means of conventional, i.e. melting-ingot casting-deformation technology. In spite of good wettability and high joint strength achieved in dozens of promising alloys, only Ti-15Cu-15Ni is now widely used as a brazing filler metal for high service temperature. Up until now this material could not be produced as a homogeneous foil and is instead applied as a clad strip consisting of three separate metallic layers.

  1. Low-energy gamma ray inspection of brazed aluminum joints

    NASA Technical Reports Server (NTRS)

    Brown, J. A.

    1967-01-01

    Americium 241 serves as a suitable radioisotope /gamma ray source/ and exposure probe for radiographic inspection of brazed aluminum joints in areas of limited accessibility. The powdered isotope is contained in a sealed capsule mounted at the end of a spring-loaded pushrod in the probe assembly.

  2. Identification of Rab41/6d Effectors Provides an Explanation for the Differential Effects of Rab41/6d and Rab6a/a' on Golgi Organization

    PubMed Central

    Liu, Shijie; Majeed, Waqar; Kudlyk, Tetyana; Lupashin, Vladimir; Storrie, Brian

    2016-01-01

    Unexpectedly, members of the Rab VI subfamily exhibit considerable variation in their effects on Golgi organization and trafficking. By fluorescence microscopy, neither depletion nor overexpression of the GDP-locked form of Rab6a/a', the first trans Golgi-associated Rab protein discovered, affects Golgi ribbon organization while, on the other hand, both Rab41/6d depletion and overexpression of GDP-locked form cause Golgi fragmentation into a cluster of punctate elements, suggesting that Rab41/6d has an active role in maintenance of Golgi ribbon organization. To establish a molecular basis for these differences, we screened for Rab41/6d interacting proteins by yeast two-hybrid assay. 155 non-repetitive hits were isolated and sequenced, and after searching in NCBI database, 102 different proteins and protein fragments were identified. None of these hits overlapped with any published Rab6a/a' effector. Eight putative Rab41 interactors involved in membrane trafficking were found. Significantly, these exhibited a preferential interaction with GTP- vs. GDP-locked Rab41/6d. Of the 8 hits, the dynactin 6, syntaxin 8, and Kif18A plasmids were the only ones expressing the full-length protein. Hence, these 3 proteins were selected for further study. We found that depletion of dynactin 6 or syntaxin 8, but not Kif18A, resulted in a fragmented Golgi apparatus that displayed a Rab41/6d knockdown phenotype, i.e., the Golgi apparatus was disrupted into a cluster of punctate Golgi elements. Co-immunoprecipation experiments verified that the interaction of dynactin 6 and syntaxin 8 with GTP-locked Rab41/6d was stronger than that with wild type Rab41/6d and least with the GDP-locked form. In contrast, co-immunoprecipitation interaction with Rab6a was greatest with the GDP-locked Rab6a, suggestive of a non-physiological interaction. In conclusion, we suggest that dynactin 6, a subunit of dynactin complex, the minus-end-directed, dynein motor, provides a sufficient molecular basis to

  3. Identification of Rab41/6d Effectors Provides an Explanation for the Differential Effects of Rab41/6d and Rab6a/a' on Golgi Organization.

    PubMed

    Liu, Shijie; Majeed, Waqar; Kudlyk, Tetyana; Lupashin, Vladimir; Storrie, Brian

    2016-01-01

    Unexpectedly, members of the Rab VI subfamily exhibit considerable variation in their effects on Golgi organization and trafficking. By fluorescence microscopy, neither depletion nor overexpression of the GDP-locked form of Rab6a/a', the first trans Golgi-associated Rab protein discovered, affects Golgi ribbon organization while, on the other hand, both Rab41/6d depletion and overexpression of GDP-locked form cause Golgi fragmentation into a cluster of punctate elements, suggesting that Rab41/6d has an active role in maintenance of Golgi ribbon organization. To establish a molecular basis for these differences, we screened for Rab41/6d interacting proteins by yeast two-hybrid assay. 155 non-repetitive hits were isolated and sequenced, and after searching in NCBI database, 102 different proteins and protein fragments were identified. None of these hits overlapped with any published Rab6a/a' effector. Eight putative Rab41 interactors involved in membrane trafficking were found. Significantly, these exhibited a preferential interaction with GTP- vs. GDP-locked Rab41/6d. Of the 8 hits, the dynactin 6, syntaxin 8, and Kif18A plasmids were the only ones expressing the full-length protein. Hence, these 3 proteins were selected for further study. We found that depletion of dynactin 6 or syntaxin 8, but not Kif18A, resulted in a fragmented Golgi apparatus that displayed a Rab41/6d knockdown phenotype, i.e., the Golgi apparatus was disrupted into a cluster of punctate Golgi elements. Co-immunoprecipation experiments verified that the interaction of dynactin 6 and syntaxin 8 with GTP-locked Rab41/6d was stronger than that with wild type Rab41/6d and least with the GDP-locked form. In contrast, co-immunoprecipitation interaction with Rab6a was greatest with the GDP-locked Rab6a, suggestive of a non-physiological interaction. In conclusion, we suggest that dynactin 6, a subunit of dynactin complex, the minus-end-directed, dynein motor, provides a sufficient molecular basis to

  4. Identification of an immune-regulated phagosomal Rab cascade in macrophages

    PubMed Central

    Pei, Gang; Repnik, Urska; Griffiths, Gareth; Gutierrez, Maximiliano Gabriel

    2014-01-01

    ABSTRACT Interferon-γ (IFN-γ) has been shown to regulate phagosome trafficking and function in macrophages, but the molecular mechanisms involved are poorly understood. Here, we identify Rab20 as part of the machinery by which IFN-γ controls phagosome maturation. We found that IFN-γ stimulates the association of Rab20 with early phagosomes in macrophages. By using imaging of single phagosomes in live cells, we found that Rab20 induces an early delay in phagosome maturation and extends the time for which Rab5a and phosphatidylinositol 3-phosphate (PI3P) remain associated with phagosomes. Moreover, Rab20 depletion in macrophages abrogates the delay in phagosome maturation induced by IFN-γ. Finally, we demonstrate that Rab20 interacts with the Rab5a guanine nucleotide exchange factor Rabex-5 (also known as RABGEF1) and that Rab20 knockdown impairs the IFN-γ-dependent recruitment of Rabex-5 and Rab5a into phagosomes. Taken together, here, we uncover Rab20 as a key player in the Rab cascade by which IFN-γ induces a delay in phagosome maturation in macrophages. PMID:24569883

  5. [Alterations in recruitment and activation of Rab proteins during mycobacterial infection].

    PubMed

    Castaño, Diana; Rojas, Mauricio

    2010-01-01

    At the phagosome level, Mycobacterium spp. alters activation and recruitment of several "Ras gene from rat brain" proteins, commonly known as Rab. Mycobacterial phagosomes have a greater and sustained expression of Rab5, Rab11, Rab14 and Rab22a, and lowered or no expression of Rab7, Rab9 and Rab6. This correlates with increased fusion of the phagosomes with early and recycling endosomes acquiring some features of early phagosomes, allowing the bacteria to gain access to nutrients and preventing the activation of anti-mycobacterial mechanisms. The expression of constitutively active mutants of Rab from the early stage endosomes prevents the maturation of phagosomes containing latex beads or heat-inactivated mycobacteria. Silencing of these mutants by interference RNA or dominant negative forms induces the maturation of mycobacterial phagosomes. The mechanisms have not been established by which mycobacteria alter the expression of these GTPases and thereby shift the phagolysosomal maturation. The problem can be explained by alterations in the recruitment of proteins that interact with Rab, such as phosphoinositide 3-kinases and early endosomal antigen 1. Identifying the mechanisms used by Mycobacterium spp. to disrupt the cycle of Rab activation will be essential to understand the pathophysiology of mycobacterial infections and usefully to potential drug targets.

  6. Brazeability of aluminum in vacuum-nitrogen partial-pressure atmosphere brazing

    SciTech Connect

    Hattori, T.; Sakai, S.; Sakamoto, A.; Fujiwara, C. )

    1994-10-01

    In vacuum brazing, Al-10% Si-1.5% Mg filler metal is used. The filler metal is melted and magnesium in the filler metal evaporates actively. The magnesium gas is the effective getter of contaminants such as H[sub 2]O and O[sub 2], which form an oxide film on the surface of aluminum alloys, lowering brazeability. Volatile elements also evaporate and material properties change in high-vacuum brazing. The vapor pressure of zinc in the Al-Zn alloy is high because zinc is a volatile element, but Al-Zn alloy does not melt at the brazing temperature, which is approximately 873 K and zinc does not evaporate actively compared with magnesium. However, evaporation of volatile elements and change in material properties can be minimized in vacuum-nitrogen partial-pressure atmosphere brazing, and Al-Zn alloy may be used as a sacrificial alloy in products made with aluminum alloys. In this study, brazeability in vacuum-nitrogen partial-pressure atmosphere was investigated using T-joints with horizontal Al-Mn or Al-Zn alloy sheet and vertical A4004 clad A3003 alloy brazing sheet. Specimens were brazed over a wide range of brazing pressures and N[sub 2] carrier gas flow rates. The brazing temperature and brazing time were 873 K (600 C) and 5 minutes, respectively. Gas contaminants in brazing atmospheres were measured using a quadruple mass spectrometer.

  7. Wetting and spreading behavior of molten brazing filler metallic alloys on metallic substrate

    NASA Astrophysics Data System (ADS)

    Kogi, Satoshi; Kajiura, Tetsurou; Hanada, Yukiakira; Miyazawa, Yasuyuki

    2014-08-01

    Wetting and spreading of molten brazing filler material are important factors that influence the brazing ability of a joint to be brazed. Several investigations into the wetting ability of a brazing filler alloy and its surface tension in molten state, in addition to effects of brazing time and temperature on the contact angle, have been carried out. In general, dissimilar-metals brazing technology and high-performance brazed joint are necessities for the manufacturing field in the near future. Therefore, to address this requirement, more such studies on wetting and spreading of filler material are required for a deeper understanding. Generally, surface roughness and surface conditions affect spreading of molten brazing filler material during brazing. Wetting by and interfacial reactions of the molten brazing filler material with the metallic substrate, especially, affect strongly the spreading of the filler material. In this study, the effects of surface roughness and surface conditions on the spreading of molten brazing filler metallic alloys were investigated. Ag-(40-x)Cu-xIn and Ag- (40-x)Cu-xSn (x=5, 10, 15, 20, 25) alloys were used as brazing filler materials. A mild-steel square plate (S45C (JIS); side: 30 mm; thickness: 3mm) was employed as the substrate. A few surfaces with varying roughness were prepared using emery paper. Brazing filler material and metallic base plate were first washed with acetone, and then a flux was applied to them. The filler, 50 mg, was placed on the center of the metallic base with the flux. A spreading test was performed under Ar gas using an electrically heated furnace, after which, the original spreading area, defined as the sessile drop area, and the apparent spreading area, produced by the capillary grooves, were both evaluated. It was observed that the spreading area decreased with increasing In and Sn content.

  8. Elemental composition of brazing alloys in metallic orthodontic brackets.

    PubMed

    Zinelis, Spiros; Annousaki, Olga; Eliades, Theodore; Makou, Margarita

    2004-06-01

    The aim of this study was to assess the elemental composition of the brazing alloy of representative orthodontic brackets. The brackets examined were Gemini (3M, Unitec, Monrovia, Calif), MicroLoc (GAC, Bohemia, NY), OptiMESHxrt (Ormco, Glendora, Calif), and Ultratrim (Dentarum, Ispringen, Germany). Four metallic brackets for each brand were embedded in epoxy resin and after metallographic grinding and polishing were cleaned in a water ultrasonic bath. Scanning electron microscopy and energy-dispersive x-ray microanalysis (EDS) were used to assess the quantitative composition of the brazing alloy. Four EDS spectra were collected for each brazing alloy, and the mean value and standard deviation for the concentration of each element were calculated. The elemental composition of the brazing alloys was determined as follows (percent weight): Gemini: Ni = 83.98 +/- 1.02, Si = 6.46 +/- 0.37, Fe = 5.90 +/- 0.93, Cr = 3.52 +/- 0.34; MicroLoc: Ag = 42.82 +/- 0.18, Au = 32.14 +/- 0.65, Cu = 24.53 +/- 0.26, Mg = 1.12 +/- 0.33; OptiMESHxrt: Au = 67.79 +/- 0.97, Fe = 15.69 +/- 0.29, Ni = 13.01 +/- 0.93, Cr = 4.01 +/- 0.35; Ultratrim: Ag = 87.97 +/- 0.33, Cu = 10.51 +/- 0.45, Mg = 1.29 +/- 0.63, Zn = 1.13 +/- 0.24. The findings of this study showed that different brazing materials were used for the different brands, and thus different performances are expected during intraoral exposure; potential effects on the biological properties also are discussed.

  9. Rab Family Proteins Regulate the Endosomal Trafficking and Function of RGS4*

    PubMed Central

    Bastin, Guillaume; Heximer, Scott P.

    2013-01-01

    RGS4, a heterotrimeric G-protein inhibitor, localizes to plasma membrane (PM) and endosomal compartments. Here, we examined Rab-mediated control of RGS4 internalization and recycling. Wild type and constitutively active Rab5 decreased RGS4 PM levels while increasing its endosomal targeting. Rab5, however, did not appreciably affect the PM localization or function of the M1 muscarinic receptor (M1R)/Gq signaling cascade. RGS4-containing endosomes co-localized with subsets of Rab5-, transferrin receptor-, and Lamp1/Lysotracker-marked compartments suggesting RGS4 traffics through PM recycling or acidified endosome pathways. Rab7 activity promoted TGN association, whereas Rab7(dominant negative) trapped RGS4 in late endosomes. Furthermore, RGS4 was found to co-localize with an endosomal pool marked by Rab11, the protein that mediates recycling/sorting of proteins to the PM. The Cys-12 residue in RGS4 appeared important for its Rab11-mediated trafficking to the PM. Rab11(dominant negative) decreased RGS4 PM levels and increased the number of RGS4-containing endosomes. Inhibition of Rab11 activity decreased RGS4 function as an inhibitor of M1R activity without affecting localization and function of the M1R/Gq signaling complex. Thus, both Rab5 activation and Rab11 inhibition decreased RGS4 function in a manner that is independent from their effects on the localization and function of the M1R/Gq signaling complex. This is the first study to implicate Rab GTPases in the intracellular trafficking of an RGS protein. Thus, Rab GTPases may be novel molecular targets for the selective regulation of M1R-mediated signaling via their specific effects on RGS4 trafficking and function. PMID:23733193

  10. Comprehensive analysis reveals dynamic and evolutionary plasticity of Rab GTPases and membrane traffic in Tetrahymena thermophila.

    PubMed

    Bright, Lydia J; Kambesis, Nichole; Nelson, Scott Brent; Jeong, Byeongmoon; Turkewitz, Aaron P

    2010-10-14

    Cellular sophistication is not exclusive to multicellular organisms, and unicellular eukaryotes can resemble differentiated animal cells in their complex network of membrane-bound structures. These comparisons can be illuminated by genome-wide surveys of key gene families. We report a systematic analysis of Rabs in a complex unicellular Ciliate, including gene prediction and phylogenetic clustering, expression profiling based on public data, and Green Fluorescent Protein (GFP) tagging. Rabs are monomeric GTPases that regulate membrane traffic. Because Rabs act as compartment-specific determinants, the number of Rabs in an organism reflects intracellular complexity. The Tetrahymena Rab family is similar in size to that in humans and includes both expansions in conserved Rab clades as well as many divergent Rabs. Importantly, more than 90% of Rabs are expressed concurrently in growing cells, while only a small subset appears specialized for other conditions. By localizing most Rabs in living cells, we could assign the majority to specific compartments. These results validated most phylogenetic assignments, but also indicated that some sequence-conserved Rabs were co-opted for novel functions. Our survey uncovered a rare example of a nuclear Rab and substantiated the existence of a previously unrecognized core Rab clade in eukaryotes. Strikingly, several functionally conserved pathways or structures were found to be associated entirely with divergent Rabs. These pathways may have permitted rapid evolution of the associated Rabs or may have arisen independently in diverse lineages and then converged. Thus, characterizing entire gene families can provide insight into the evolutionary flexibility of fundamental cellular pathways.

  11. Rab family proteins regulate the endosomal trafficking and function of RGS4.

    PubMed

    Bastin, Guillaume; Heximer, Scott P

    2013-07-26

    RGS4, a heterotrimeric G-protein inhibitor, localizes to plasma membrane (PM) and endosomal compartments. Here, we examined Rab-mediated control of RGS4 internalization and recycling. Wild type and constitutively active Rab5 decreased RGS4 PM levels while increasing its endosomal targeting. Rab5, however, did not appreciably affect the PM localization or function of the M1 muscarinic receptor (M1R)/Gq signaling cascade. RGS4-containing endosomes co-localized with subsets of Rab5-, transferrin receptor-, and Lamp1/Lysotracker-marked compartments suggesting RGS4 traffics through PM recycling or acidified endosome pathways. Rab7 activity promoted TGN association, whereas Rab7(dominant negative) trapped RGS4 in late endosomes. Furthermore, RGS4 was found to co-localize with an endosomal pool marked by Rab11, the protein that mediates recycling/sorting of proteins to the PM. The Cys-12 residue in RGS4 appeared important for its Rab11-mediated trafficking to the PM. Rab11(dominant negative) decreased RGS4 PM levels and increased the number of RGS4-containing endosomes. Inhibition of Rab11 activity decreased RGS4 function as an inhibitor of M1R activity without affecting localization and function of the M1R/Gq signaling complex. Thus, both Rab5 activation and Rab11 inhibition decreased RGS4 function in a manner that is independent from their effects on the localization and function of the M1R/Gq signaling complex. This is the first study to implicate Rab GTPases in the intracellular trafficking of an RGS protein. Thus, Rab GTPases may be novel molecular targets for the selective regulation of M1R-mediated signaling via their specific effects on RGS4 trafficking and function.

  12. Low-melting-point titanium-base brazing alloys. Part 2: Characteristics of brazing Ti-21Ni-14Cu on Ti-6Al-4V substrate

    SciTech Connect

    Chang, E.; Chen, C.H.

    1997-12-01

    Filler metal of a low-melting-point (917 C) Ti-21Ni-14Cu was brazed onto the substrate of Ti-6Al-4V alloy at 960 C for 2, 4, and 8 h to investigate the microstructural evolution and electrochemical characteristics of the brazed metal as a function of the period of brazing time. Optical microscopy, scanning and transmission electron microscopy, and x-ray diffractometry were used to characterize the microstructure and phase of the brazed metal; also, the potentiostat was used for corrosion study. Experimental results indicate that diffusion of copper and nickel from the filler metal into the equiaxed {alpha} plus intergranular {beta} structure of Ti-6Al-4V substrate causes the lamellar Widmanstaetten structure to form. The intermetallic Ti{sub 2}Ni phase existing in the prior filler metal diminishes, while the Ti{sub 2}Cu phase can be identified for the metal brazed at 960 C for 2 h, but the latter phase decreases with time. Advantage might be taken from the evidence of faster diffusion of nickel than copper along the {beta} phase to the substrate. In deaerated Hank`s solution, corrosion potential, corrosion current density, and critical potential for active-to-passive transition decrease while the passivation range broadens with the period of brazing time. However, all the brazed metals, immersed for different periods in oxygen-saturated Hank`s solution, show similar corrosion behavior, irrespective of the brazing time.

  13. Low-melting-point titanium-base brazing alloys—part 2: Characteristics of brazing Ti-21Ni-14Cu on Ti-6Al-4v substrate

    NASA Astrophysics Data System (ADS)

    Chang, E.; Chen, C.-H.

    1997-12-01

    Filler metal of a low-melting-point (917 °C) Ti-21Ni-14Cu was brazed onto the substrate of Ti-6Al-4V alloy at 960 °C for 2,4, and 8 h to investigate the microstructural evolution and electrochemical characteristics of the brazed metal as a function of the period of brazing time. Optical microscopy, scanning and transmission electron microscopy, and x-ray diffractometry were used to characterize the microstructure and phase of the brazed metal; also, the potentiostat was used for corrosion study. Experimental results indicate that diffusion of copper and nickel from the filler metal into the equiaxed a plus intergranular β structure of Ti-6Al-4V substrate causes the lamellar Widmanstätten structure to form. The intermetallic Ti2Ni phase existing in the prior filler metal diminishes, while the Ti2Cu phase can be identified for the metal brazed at 960 °C for 2 h, but the latter phase decreases with time. Advantage might be taken from the evidence of faster diffusion of nickel than copper along the β phase to the substrate. In deaerated Hank’s solution, corrosion potential, corrosion current density, and critical potential for active-to-passive transition decrease while the passivation range broadens with the period of brazing time. However, all the brazed metals, immersed for different periods in oxygen-saturated Hank’s solution, show similar corrosion behavior, irrespective of the brazing time.

  14. Endocytic turnover of Rab8 controls cell polarization

    PubMed Central

    Vidal-Quadras, Maite; Holst, Mikkel R.; Larsson, Elin; Hachimi, Mariam; Yau, Wai-Lok; Peränen, Johan; Martín-Belmonte, Fernando

    2017-01-01

    ABSTRACT Adaptation of cell shape and polarization through the formation and retraction of cellular protrusions requires balancing of endocytosis and exocytosis combined with fine-tuning of the local activity of small GTPases like Rab8. Here, we show that endocytic turnover of the plasma membrane at protrusions is directly coupled to surface removal and inactivation of Rab8. Removal is induced by reduced membrane tension and mediated by the GTPase regulator associated with focal adhesion kinase-1 (GRAF1, also known as ARHGAP26), a regulator of clathrin-independent endocytosis. GRAF1-depleted cells were deficient in multi-directional spreading and displayed elevated levels of GTP-loaded Rab8, which was accumulated at the tips of static protrusions. Furthermore, GRAF1 depletion impaired lumen formation and spindle orientation in a 3D cell culture system, indicating that GRAF1 activity regulates polarity establishment. Our data suggest that GRAF1-mediated removal of Rab8 from the cell surface restricts its activity during protrusion formation, thereby facilitating dynamic adjustment of the polarity axis. PMID:28137756

  15. Endocytic turnover of Rab8 controls cell polarization.

    PubMed

    Vidal-Quadras, Maite; Holst, Mikkel R; Francis, Monika K; Larsson, Elin; Hachimi, Mariam; Yau, Wai-Lok; Peränen, Johan; Martín-Belmonte, Fernando; Lundmark, Richard

    2017-03-15

    Adaptation of cell shape and polarization through the formation and retraction of cellular protrusions requires balancing of endocytosis and exocytosis combined with fine-tuning of the local activity of small GTPases like Rab8. Here, we show that endocytic turnover of the plasma membrane at protrusions is directly coupled to surface removal and inactivation of Rab8. Removal is induced by reduced membrane tension and mediated by the GTPase regulator associated with focal adhesion kinase-1 (GRAF1, also known as ARHGAP26), a regulator of clathrin-independent endocytosis. GRAF1-depleted cells were deficient in multi-directional spreading and displayed elevated levels of GTP-loaded Rab8, which was accumulated at the tips of static protrusions. Furthermore, GRAF1 depletion impaired lumen formation and spindle orientation in a 3D cell culture system, indicating that GRAF1 activity regulates polarity establishment. Our data suggest that GRAF1-mediated removal of Rab8 from the cell surface restricts its activity during protrusion formation, thereby facilitating dynamic adjustment of the polarity axis.

  16. Development and quality evaluation of value added instant rab mixes.

    PubMed

    Mogra, Renu; Choudhry, Maya

    2014-06-01

    Rab is a traditional Rajasthani preparation prepared by boiling maize grits and or flour in buttermilk for 2-3 h. It is sour in taste with curd and cumin seed flavour and has thick soup like consistency. Two instant mixes of rab were developed using two types of curds prepared from two different culture combination (i) S. lactis + S. thermophilus + S. diacetylactis and (ii) L. bulgaricus + S. diacetylactis. Processed greengram dhal and spinach leaves powder were added to the mixes for their value addition. In all, six mixes were developed from two types of curd. The mixes were analysed for nutritional composition at zero month and sensory characteristics and shelf life was evaluated for 4 months. The rab mixes provided moisture 4.0-5.5 g; protein 15.1-17.6 g; carbohydrate 62.3-70.29 g; energy 357-394 kcal; fat 4.4-6.1 g; iron 3.15-3.89 mg and calcium 346-386 mg per 100 g of mixes. There was an increase in protein, ash, fiber and iron content of mixes through value addition. Sensory evaluation of rab reconstituted from mixes revealed that the scores ranged between 7 and 8 depicting that rab was liked very much. The mixes were free from coliforms and S.aureus bacteria and the counts for TVC, yeast and molds were within safe limits during storage of 4 months at 28-30°C and RH 55-65%.

  17. Evaluation of Superplastic Forming and Weld-brazing for Fabrication of Titanium Compression Panels

    NASA Technical Reports Server (NTRS)

    Royster, D. M.; Bales, T. T.; Davis, R. C.

    1985-01-01

    The two titanium processing procedures, superplastic forming and weld brazing, are successfully combined to fabricate titanium skin stiffened structural panels. Stiffeners with complex shapes are superplastically formed using simple tooling. These stiffeners are formed to the desired configuration and required no additional sizing or shaping following removal from the mold. The weld brazing process by which the stiffeners are attached to the skins utilize spot welds to maintain alignment and no additional tooling is required for brazing. The superplastic formed/weld brazed panels having complex shaped stiffeners develop up to 60 percent higher buckling strengths than panels with conventional shaped stiffeners. The superplastic forming/weld brazing process is successfully scaled up to fabricate full size panels having multiple stiffeners. The superplastic forming/weld brazing process is also successfully refined to show its potential for fabricating multiple stiffener compression panels employing unique stiffener configurations for improved structural efficiency.

  18. Preliminary study on pressure brazing and diffusion welding of Nb-1Zr to Inconel 718

    NASA Technical Reports Server (NTRS)

    Moore, T. J.

    1990-01-01

    Future space power systems may include Nb-1Zr/Inconel 718 dissimilar metal joints for operation at 1000 K for 60,000 h. The serviceability of pressure-brazed and diffusion-welded joints was investigated. Ni-based metallic glass foil filler metals were used for brazing. Ni and Fe foils were used as diffusion welding inter-layers. Joint soundness was determined by metallographic examination in the as-brazed and as-welded condition, after aging at 1000 K, and after thermal cycling. Brazed joints thermally cycled in the as-brazed condition and diffusion-welded joints were unsatisfactory because of cracking problems. Brazed joints may meet the service requirements if the joints are aged at 1000 K prior to thermal cycling.

  19. Dissimilar joint characteristics of SiC and WC-Co alloy by laser brazing

    NASA Astrophysics Data System (ADS)

    Nagatsuka, K.; Sechi, Y.; Nakata, K.

    2012-08-01

    SiC and WC-Co alloys were joined by laser brazing with an active braze metal. The braze metal based on eutectic Ag-Cu alloy with additional Ti as an active element ranging from 0 to 2.8 mass% was sandwiched by the SiC block and WC-Co alloy plate. The brazing was carried out by selective laser beam irradiation on the WC-Co alloy plate. The content of Ti in the braze metal was required to exceed 0.6 mass% in order to form a brazed joint with a measurable shear strength. The shear strength increased with increasing Ti content up to 2.3 mass%Ti and decreased with a higher content.

  20. Rab 11 regulates constitutive dopamine transporter trafficking and function in N2A neuroblastoma cells.

    PubMed

    Furman, Cheryse A; Lo, Charles B; Stokes, Stephanie; Esteban, Jose A; Gnegy, Margaret E

    2009-09-29

    The dopamine transporter (DAT) is a crucial regulator of dopaminergic neurotransmission which undergoes constitutive and substrate-mediated trafficking to and from the membrane. Although, considerable research has been done to elucidate the regulation of substrate-stimulated DAT trafficking, less is known about which trafficking proteins are involved in constitutive DAT trafficking. Rab proteins are GTPases known to regulate the trafficking of proteins to and from specific endocytic compartments. Rabs 8 and 11, in particular, are involved in trafficking proteins from intracellular compartments to the plasma membrane. In this study, we sought to determine whether Rabs 8 and 11 would modulate DAT activity and trafficking in N2A neuroblastoma cells. We used Rab mutations known to confer constitutively active or dominant negative activity of these proteins to investigate the role of Rab activity in constitutive DAT trafficking and function. We found that constitutively active Rab 11 upregulates DAT function and surface expression while neither the constitutively active nor the dominant negative mutant of Rab 8 had any effect on DA uptake. Furthermore, immunofluorescence experiments revealed that dominant negative Rab 11 overexpression results in decreased surface DAT indicating a necessary function of Rab 11 in DAT trafficking to the plasma membrane. These data show for the first time a functional role of Rab proteins in the constitutive recycling of DAT to the plasma membrane.

  1. bMERB domains are bivalent Rab8 family effectors evolved by gene duplication

    PubMed Central

    Rai, Amrita; Oprisko, Anastasia; Campos, Jeremy; Fu, Yangxue; Friese, Timon; Itzen, Aymelt; Goody, Roger S; Gazdag, Emerich Mihai; Müller, Matthias P

    2016-01-01

    In their active GTP-bound form, Rab proteins interact with proteins termed effector molecules. In this study, we have thoroughly characterized a Rab effector domain that is present in proteins of the Mical and EHBP families, both known to act in endosomal trafficking. Within our study, we show that these effectors display a preference for Rab8 family proteins (Rab8, 10, 13 and 15) and that some of the effector domains can bind two Rab proteins via separate binding sites. Structural analysis allowed us to explain the specificity towards Rab8 family members and the presence of two similar Rab binding sites that must have evolved via gene duplication. This study is the first to thoroughly characterize a Rab effector protein that contains two separate Rab binding sites within a single domain, allowing Micals and EHBPs to bind two Rabs simultaneously, thus suggesting previously unknown functions of these effector molecules in endosomal trafficking. DOI: http://dx.doi.org/10.7554/eLife.18675.001 PMID:27552051

  2. [Isolation, phylogenetic analysis and developmental expression parttern of AmphiRab23b in amphioxus].

    PubMed

    Li, Jian-Wei; Lin, Yu-Shuang; Chen, Dong-Yan; Zhang, Hong-Wei

    2009-12-01

    The hedgehog (Hh) pathway plays an important role during the embryonic development and is related to the progression of cancers. Rab23 is a crucial functional molecule in Hh pathway. However, there is no report about amphioxus Rab23 up to now except the annotations of two isoforms in the genome of Florida lancelet (Branchiostoma floridae). Here a 2062 bp full-length cDNA sequence of the Rab23, AmphiRab23b, was isolated from Chinese amphioxus (Branchiostoma belcheri), which included the UTRs and an open reading frame of 714 bp, encoding a protein of 237 amino acids. Phylogenetic analysis suggested that AmphiRab23b falled outside the vertebrate clade. But sequence analysis indicated that this putative AmphiRab23b protein contained a specific Rab23_lke domain, which implied that Rab23 gene was functional conservative during evolution. And its developmental expression pattern showed that AmphiRab23b was expressed in the differentiating neural plate and alimentary canal, as the same as the expression pattern of the homologous vertebrate genes, which suggested that AmphiRab23b may function in the development of nervous system and alimentary canal.

  3. Yersinia pestis Requires Host Rab1b for Survival in Macrophages

    PubMed Central

    Connor, Michael G.; Pulsifer, Amanda R.; Price, Christopher T.; Abu Kwaik, Yousef; Lawrenz, Matthew B.

    2015-01-01

    Yersinia pestis is a facultative intracellular pathogen that causes the disease known as plague. During infection of macrophages Y. pestis actively evades the normal phagosomal maturation pathway to establish a replicative niche within the cell. However, the mechanisms used by Y. pestis to subvert killing by the macrophage are unknown. Host Rab GTPases are central mediators of vesicular trafficking and are commonly targeted by bacterial pathogens to alter phagosome maturation and killing by macrophages. Here we demonstrate for the first time that host Rab1b is required for Y. pestis to effectively evade killing by macrophages. We also show that Rab1b is specifically recruited to the Yersinia containing vacuole (YCV) and that Y. pestis is unable to subvert YCV acidification when Rab1b expression is knocked down in macrophages. Furthermore, Rab1b knockdown also altered the frequency of association between the YCV with the lysosomal marker Lamp1, suggesting that Rab1b recruitment to the YCV directly inhibits phagosome maturation. Finally, we show that Rab1b knockdown also impacts the pH of the Legionella pneumophila containing vacuole, another pathogen that recruits Rab1b to its vacuole. Together these data identify a novel role for Rab1b in the subversion of phagosome maturation by intracellular pathogens and suggest that recruitment of Rab1b to the pathogen containing vacuole may be a conserved mechanism to control vacuole pH. PMID:26495854

  4. Interferon-γ-inducible Rab20 regulates endosomal morphology and EGFR degradation in macrophages.

    PubMed

    Pei, Gang; Schnettger, Laura; Bronietzki, Marc; Repnik, Urska; Griffiths, Gareth; Gutierrez, Maximiliano Gabriel

    2015-09-01

    Little is known about the molecular players that regulate changes in the endocytic pathway during immune activation. Here we investigate the role of Rab20 in the endocytic pathway during activation of macrophages. Rab20 is associated with endocytic structures, but the function of this Rab GTPase in the endocytic pathway remains poorly characterized. We find that in macrophages, Rab20 expression and endosomal association significantly increase after interferon-γ (IFN-γ) treatment. Moreover, IFN-γ and Rab20 expression induce a dramatic enlargement of endosomes. These enlarged endosomes are the result of homotypic fusion promoted by Rab20 expression. The expression of Rab20 or the dominant-negative mutant Rab20T19N does not affect transferrin or dextran 70 kDa uptake. However, knockdown of Rab20 accelerates epidermal growth factor (EGF) trafficking to LAMP-2-positive compartments and EGF receptor degradation. Thus this work defines a function for Rab20 in the endocytic pathway during immune activation of macrophages.

  5. Phosphorylation of Rab11-FIP2 regulates polarity in MDCK cells

    PubMed Central

    Lapierre, Lynne A.; Avant, Kenya M.; Caldwell, Cathy M.; Oztan, Asli; Apodaca, Gerard; Knowles, Byron C.; Roland, Joseph T.; Ducharme, Nicole A.; Goldenring, James R.

    2012-01-01

    The Rab11 effector Rab11-family interacting protein 2 (Rab11-FIP2) regulates transcytosis through its interactions with Rab11a and myosin Vb. Previous studies implicated Rab11-FIP2 in the establishment of polarity in Madin–Darby canine kidney (MDCK) cells through phosphorylation of Ser-227 by MARK2. Here we examine the dynamic role of Rab11-FIP2 phosphorylation on MDCK cell polarity. Endogenous Rab11-FIP2 phosphorylated on Ser-227 coalesces on vesicular plaques during the reestablishment of polarity after either monolayer wounding or calcium switch. Whereas expression of the nonphosphorylatable Rab11-FIP2(S227A) elicits a loss in lumen formation in MDCK cell cysts grown in Matrigel, the putative pseudophosphorylated Rab11-FIP2(S227E) mutant induces the formation of cysts with multiple lumens. On permeable filters, Rab11-FIP2(S227E)–expressing cells exhibit alterations in the composition of both the adherens and tight junctions. At the adherens junction, p120 catenin and K-cadherin are retained, whereas the majority of the E-cadherin is lost. Although ZO-1 is retained at the tight junction, occludin is lost and the claudin composition is altered. Of interest, the effects of Rab11-FIP2 on cellular polarity did not involve myosin Vb or Rab11a. These results indicate that Ser-227 phosphorylation of Rab11-FIP2 regulates the composition of both adherens and tight junctions and is intimately involved in the regulation of polarity in epithelial cells. PMID:22553350

  6. The use of Ni-Cr-Si-Be filler metals for brazing of stainless steels

    NASA Astrophysics Data System (ADS)

    Ivannikov, A.; Fedotov, V.; Suchkov, A.; Penyaz, M.; Fedotov, I.; Tarasov, B.

    2016-04-01

    Nanocrystalline ribbon filler metal-alloys of system Ni-Cr-Si-Be are produced by the rapidly quenching of the melt method. By these filler metals carried out hight temperature vacuum brazing of austenitic steels (12Kh18N10T and Kh18N8G2) and austenitic-ferritic class EI-811 (12Kh21N5T). The basic laws of structure-phase state foundation of brazed joints are determined, features of the interaction of the molten filler metal to the brazed materials are identified, the optimal temperature and time parameters of the brazing process are determined.

  7. Effect of environment on the quality of aluminum joints brazed in partial nitrogen atmospheres

    SciTech Connect

    Pai, D.M.; Zhong, G.; Shafiq, P.; Badgley, S.

    1995-06-01

    The vacuum brazing of aluminum is an established process for producing parts such as radiators and heater cores used in automobiles. The process forms connecting fillets between component parts by the melting and flow of aluminum-silicon alloy cladding. The operating procedures developed for vacuum brazing are largely empirical, and rely on process and materials control. Past studies have examined the effects of factors such as moisture, heating rate, the wetting and flow characteristics of filler alloy and the role of the magnesium promoter in the vacuum brazing process. While vacuum brazing has traditionally been done under high vacuum conditions, it is economically desirable to have less stringent vacuum requirements. This study focuses on the effect of environment on the quality of aluminum joints brazed in an atmosphere where a high vacuum has been produced and then backfilled partially with nitrogen. Microscopic examination of the braze quality together with an analysis of the vacuum furnace atmosphere reveals a possible correlation between gaseous impurity levels in the brazing environment and the quality of the brazed joint. This paper reports the results of this study and provides data on limits of environmental impurities that can be tolerated by the process. This data can eventually be used for intelligent computer control, and thereby optimization, of the vacuum brazing process.

  8. Infrared Brazing Ti50Ni50 and Invar Using Ag-Based Filler Foils

    NASA Astrophysics Data System (ADS)

    Shiue, R. K.; Chang, Y. H.; Wu, S. K.

    2013-10-01

    Infrared brazing Ti50Ni50 and Invar using BAg-8 and Cusil-ABA foils was investigated. The Ag-Cu eutectic matrix dominates both brazed joints. The maximum shear strengths of the brazed joints using BAg-8 and Cusil-ABA fillers are 158 and 249 MPa. Failure of interfacial Fe2Ti/Ni3Ti reaction layers is responsible for the BAg-8 joint. In contrast, the Cusil-ABA brazed joint is fractured along the interfacial Fe2Ti intermetallic compound. Both fractographs are characterized with cleavage dominated fracture.

  9. Arabidopsis RabF1 (ARA6) Is Involved in Salt Stress and Dark-Induced Senescence (DIS)

    PubMed Central

    Yin, Congfei; Karim, Sazzad; Zhang, Hongsheng; Aronsson, Henrik

    2017-01-01

    Arabidopsis small GTPase RabF1 (ARA6) functions in endosomal vesicle transport and may play a crucial role in recycling and degradation of molecules, thus involved in stress responses. Here we have reported that complementary overexpression lines RabF1OE (overexpression), GTPase mutants RabF1Q93L (constitutively active) and RabF1S47N (dominant negative) lines show longer root growth than wild-type, rabF1 knockout and N-myristoylation deletion (Δ1−29, N-terminus) complementary overexpression mutant plants under salt induced stress, which indicates that N-myristoylation of RabF1 is indispensable for salt tolerance. Moreover, RabF1 is highly expressed during senescence and RabF1OE lines were more tolerant of dark-induced senescence (DIS) than wild-type and rabF1. PMID:28157156

  10. RAB-5- and RAB-11-dependent vesicle-trafficking pathways are required for plasma membrane repair after attack by bacterial pore-forming toxin.

    PubMed

    Los, Ferdinand C O; Kao, Cheng-Yuan; Smitham, Jane; McDonald, Kent L; Ha, Christine; Peixoto, Christina A; Aroian, Raffi V

    2011-02-17

    Pore-forming toxins (PFTs) secreted by pathogenic bacteria are the most common bacterial protein toxins and are important virulence factors for infection. PFTs punch holes in host cell plasma membranes, and although cells can counteract the resulting membrane damage, the underlying mechanisms at play remain unclear. Using Caenorhabditis elegans as a model, we demonstrate in vivo and in an intact epithelium that intestinal cells respond to PFTs by increasing levels of endocytosis, dependent upon RAB-5 and RAB-11, which are master regulators of endocytic and exocytic events. Furthermore, we find that RAB-5 and RAB-11 are required for protection against PFT and to restore integrity to the plasma membrane. One physical mechanism involved is the RAB-11-dependent expulsion of microvilli from the apical side of the intestinal epithelial cells. Specific vesicle-trafficking pathways thus protect cells against an attack by PFTs on plasma membrane integrity, via altered plasma membrane dynamics.

  11. Analysis of the Rab GTPase Interactome in Dendritic Cells Reveals Anti-microbial Functions of the Rab32 Complex in Bacterial Containment.

    PubMed

    Li, Yuanyuan; Wang, Yu; Zou, Liyun; Tang, Xiangyu; Yang, Yi; Ma, Li; Jia, Qingzhu; Ni, Qingshan; Liu, Siqi; Tang, Lizhang; Lin, Regina; Wong, Elizabeth; Sun, Wei; Wang, Liting; Wei, Quanfang; Ran, Haiying; Zhang, Liqun; Lian, Hengning; Huang, Wei; Wu, Yuzhang; Li, Qi-Jing; Wan, Ying

    2016-02-16

    Dendritic cells (DCs) orchestrate complex membrane trafficking through an interconnected transportation network linked together by Rab GTPases. Through a tandem affinity purification strategy and mass spectrometry, we depicted an interactomic landscape of major members of the mammalian Rab GTPase family. When complemented with imaging tools, this proteomic analysis provided a global view of intracellular membrane organization. Driven by this analysis, we investigated dynamic changes to the Rab32 subnetwork in DCs induced by L. monocytogenes infection and uncovered an essential role of this subnetwork in controlling the intracellular proliferation of L. monocytogenes. Mechanistically, Rab32 formed a persistent complex with two interacting proteins, PHB and PHB2, to encompass bacteria both during early phagosome formation and after L. monocytogenes escaped the original containment vacuole. Collectively, we have provided a functional compartmentalization overview and an organizational framework of intracellular Rab-mediated vesicle trafficking that can serve as a resource for future investigations.

  12. Prenylation of Rab8 GTPase by type I and type II geranylgeranyl transferases.

    PubMed Central

    Wilson, A L; Erdman, R A; Castellano, F; Maltese, W A

    1998-01-01

    Rab GTPases are post-translationally modified by addition of geranylgeranyl moieties to carboxyl-terminal cysteine residues. For Rab proteins ending with xxCC xCxC and CCxx motifs this modification is catalysed by geranylgeranyltransferase type II (GGTaseII), and is entirely dependent on the Rab substrate being bound to Rab escort protein (REP). Several Rab proteins contain carboxyl-terminal CaaL prenylation motifs typical of members of the Rho family, which are modified in a REP-independent manner by geranylgeranyltransferase type I (GGTaseI). The present studies show that one such Rab protein (Rab8), which ends with a CVLL motif, is uniquely able to serve as a substrate for either REP/GGTaseII or GGTaseI in cell-free assays. The modification of Rab8 by GGTaseI did not require REP, indicating that a REP-induced conformational change is not essential for exposure of the Rab carboxyl-terminal cysteine prenylation site. To determine whether one enzyme plays a predominant role in Rab8 prenylation in vivo, the incorporation of [3H]mevalonate into Rab8 was measured in human embryonal kidney 293 cells under conditions where the activity of GGTaseI, but not GGTaseII, was blocked by the peptidomimetic inhibitor GGTI-298. The GGTaseI inhibitor did not prevent prenylation of either overexpressed Myc-tagged Rab8 or endogenous Rab8, whereas prenylation of a known GGTaseI substrate with the same carboxyl-terminal motif, Cdc42Hs, was completely blocked. To rule out the possibility that the apparent prenylation of Rab8 by GGTaseII occurs only when GGTaseI activity is eliminated, metabolic labelling studies were carried out in the absence of the GGTaseI inhibitor, using a REP-binding-deficient Rab8 construct (Y78D) that cannot serve as a substrate for GGTaseII, but is indistinguishable from wild-type Rab8 as a substrate for GGTaseI. Prenylation of the Y78D mutant was reduced by 60-70% in intact cells, consistent with the conclusion that the majority of Rab8 is prenylated by the

  13. New hermetic sealing material for vacuum brazing of stainless steels

    NASA Astrophysics Data System (ADS)

    Hildebrandt, S.; Wiehl, G.; Silze, F.

    2016-03-01

    For vacuum brazing applications such as in vacuum interrupter industry Hermetic Sealing Materials (HSM) with low partial pressure are widely used. AgCu28 dominates the hermetic sealing market, as it has a very good wetting behavior on copper and metallized ceramics. Within recent decades wetting on stainless steel has become more and more important. However, today the silver content of HSMs is more in focus than in the past decades, because it has the biggest impact on the material prices. Umicore Technical Materials has developed a new copper based HSM, CuAg40Ga10. The wettability on stainless steel is significantly improved compared to AgCu28 and the total silver content is reduced by almost 44%. In this article the physical properties of the alloy and its brazed joints will be presented compared to AgCu28.

  14. Effects of service environments on aluminum-brazed titanium (ABTi)

    NASA Technical Reports Server (NTRS)

    Cotton, W. L.

    1978-01-01

    Aluminum brazed titanium (ABTi) structures were evaluated during prolonged exposure to extreme environments: elevated temperature exposure to airline service fluids, hydraulic fluid, and seawater, followed by laboratory corrosion tests. Solid-face and perforated face honeycomb sandwich panel specimens, stressed panel assemblies, and faying surface brazed joints were tested. The corrosion resistance of ABTi is satisfactory for commercial airline service. Unprotected ABTi proved inherently resistant to attack by all of the extreme service aircraft environments except: seawater at 700 K (800 F) and above, dripping phosphate ester hydraulic fluid at 505 K (450 F), and a marine environment at ambient temperature. The natural oxides and deposits present on titanium surfaces in airline service provide protection against hot salt corrosion pitting. Coatings are required to protect titanium dripping phosphate ester fluid at elevated temperatures and to protect exposed acoustic honeycomb parts against corrosion in a marine environment.

  15. Separation and Sealing of a Sample Container Using Brazing

    NASA Technical Reports Server (NTRS)

    Bar-Cohen, Yoseph; Rivellini, Tommaso P.; Wincentsen, James E.; Gershman, Robert

    2007-01-01

    A special double-wall container and a process for utilizing the container are being developed to enable (1) acquisition of a sample of material in a dirty environment that may include a biological and/or chemical hazard; (2) sealing a lid onto the inner part of the container to hermetically enclose the sample; (3) separating the resulting hermetic container from the dirty environment; and (4) bringing that hermetic container, without any biological or chemical contamination of its outer surface, into a clean environment. The process is denoted S(exp 3)B (separation, seaming, and sealing using brazing) because sealing of the sample into the hermetic container, separating the container from the dirty environment, and bringing the container with a clean outer surface into the clean environment are all accomplished simultaneously with a brazing operation.

  16. Corrosion Testing of Brazed Space Station IATCS Materials

    NASA Technical Reports Server (NTRS)

    Pohlman, Matthew J.; Varisik, Jerry; Steele, John W.; Golden, Johnny L.; Boyce, William E.; Pedley, Michael D.

    2004-01-01

    Increased nickel concentrations in the IATCS coolant prompted a study of the corrosion rates of nickel-brazed heat exchangers in the system. The testing has shown that corrosion is occurring in a silicon-rich intermetallic phase in the braze filler of coldplates and heat exchangers as the result of a decrease in the coolant pH brought about by cabin carbon dioxide permeation through polymeric flexhoses. Similar corrosion is occurring in the EMU de-ionized water loop. Certain heat exchangers and coldplates have more silicon-rich phase because of their manufacturing method, and those units produce more nickel corrosion product. Silver biocide additions did not induce pitting corrosion at silver precipitate sites.

  17. Low vapor pressure braze alloys for thermionic energy converters

    NASA Technical Reports Server (NTRS)

    Bair, V. L.

    1976-01-01

    Preliminary results in the use of some low-vapor-pressure braze alloys are reported; these are binary alloys of refractory metals (Th, Zr, Hf, Ru, Nb, Ir, Mo, Ta, Os, Re, W) with vapor pressures below 0.1 nanotorr at 1500 K or 10 microtorr at 2000 K. The melting point minima or eutectics of the alloys range from 1510 K to above 3000 K. Melting points and surface wetting on a Ta base are given. Results are presented on brazing of Ir, LaB6, Nb, Re, W, and ZrO2 (with 22 wt % Zr) into a Ta base or a Nb-1% Zr base. The results are applicable in electrode screening programs for thermionic cesium diodes.

  18. HIGH TEMPERATURE BRAZING ALLOY FOR JOINT Fe-Cr-Al MATERIALS AND AUSTENITIC AND FERRITIC STAINLESS STEELS

    DOEpatents

    Cost, R.C.

    1958-07-15

    A new high temperature brazing alloy is described that is particularly suitable for brazing iron-chromiumaluminum alloys. It consists of approximately 20% Cr, 6% Al, 10% Si, and from 1.5 to 5% phosphorus, the balance being iron.

  19. Recycling and Endosomal Sorting of Protease-activated Receptor-1 Is Distinctly Regulated by Rab11A and Rab11B Proteins*

    PubMed Central

    Grimsey, Neil J.; Coronel, Luisa J.; Cordova, Isabel Canto; Trejo, JoAnn

    2016-01-01

    Protease-activated receptor-1 (PAR1) is a G protein-coupled receptor that undergoes proteolytic irreversible activation by coagulant and anti-coagulant proteases. Given the irreversible activation of PAR1, signaling by the receptor is tightly regulated through desensitization and intracellular trafficking. PAR1 displays both constitutive and agonist-induced internalization. Constitutive internalization of PAR1 is important for generating an internal pool of naïve receptors that replenish the cell surface and facilitate resensitization, whereas agonist-induced internalization of PAR1 is critical for terminating G protein signaling. We showed that PAR1 constitutive internalization is mediated by the adaptor protein complex-2 (AP-2), whereas AP-2 and epsin control agonist-induced PAR1 internalization. However, the mechanisms that regulate PAR1 recycling are not known. In the present study we screened a siRNA library of 140 different membrane trafficking proteins to identify key regulators of PAR1 intracellular trafficking. In addition to known mediators of PAR1 endocytosis, we identified Rab11B as a critical regulator of PAR1 trafficking. We found that siRNA-mediated depletion of Rab11B and not Rab11A blocks PAR1 recycling, which enhanced receptor lysosomal degradation. Although Rab11A is not required for PAR1 recycling, depletion of Rab11A resulted in intracellular accumulation of PAR1 through disruption of basal lysosomal degradation of the receptor. Moreover, enhanced degradation of PAR1 observed in Rab11B-deficient cells is blocked by depletion of Rab11A and the autophagy related-5 protein, suggesting that PAR1 is shuttled to an autophagic degradation pathway in the absence of Rab11B recycling. Together these findings suggest that Rab11A and Rab11B differentially regulate intracellular trafficking of PAR1 through distinct endosomal sorting mechanisms. PMID:26635365

  20. Brazing SiC/SiC Composites to Metals

    NASA Technical Reports Server (NTRS)

    Steffier, Wayne S.

    2004-01-01

    Experiments have shown that active brazing alloys (ABAs) can be used to join SiC/SiC composite materials to metals, with bond strengths sufficient for some structural applications. The SiC/SiC composite coupons used in the experiments were made from polymerbased SiC fiber preforms that were chemical-vapor-infiltrated with SiC to form SiC matrices. Some of the metal coupons used in the experiments were made from 304 stainless steel; others were made from oxygen-free, high-conductivity copper. Three ABAs were chosen for the experiments: two were chosen randomly from among a number of ABAs that were on hand at the time; the third ABA was chosen because its titanium content (1.25 percent) is less than those of the other two ABAs (1.75 and 4.5 percent, respectively) and it was desired to evaluate the effect of reducing the titanium content, as described below. The characteristics of ABAs that are considered to be beneficial for the purpose of joining SiC/SiC to metal include wettability, reactivity, and adhesion to SiC-based ceramics. Prior to further development, it was verified that the three chosen ABAs have these characteristics. For each ABA, suitable vacuum brazing process conditions were established empirically by producing a series of (SiC/SiC)/ABA wetting samples. These samples were then sectioned and subjected to scanning electron microscopy (SEM) and energy-dispersive x-ray spectrometry (EDS) for analysis of their microstructures and compositions. Specimens for destructive mechanical tests were fabricated by brazing of lap joints between SiC/SiC coupons 1/8-in. (.3.2- mm) thick and, variously, stainless steel or copper tabs. The results of destructive mechanical tests and the SEM/EDS analysis were used to guide the development of a viable method of brazing the affected materials.

  1. Feasibility study of fluxless brazing cemented carbides to steel

    NASA Astrophysics Data System (ADS)

    Tillmann, W.; Sievers, N.

    2017-03-01

    One of the most important brazing processes is the joints between cemented carbides and steel for the tool industry such as in rotary drill hammers or saw blades. Even though this technique has already been used for several decades, defects in the joint can still occur and lead to quality loss. Mostly, the joining process is facilitated by induction heating and the use of a flux to enhance the wetting of the filler alloy on the surface of the steel and cemented carbide in an ambient atmosphere. However, although the use of flux enables successful joining, it also generates voids within the joint, which reduces the strength of the connection while the chemicals within the flux are toxic and polluting. In this feasibility study, a fluxless brazing process is used to examine the joint between cemented carbides and steel for the first time. For this, ultrasound is applied during induction heating to enable the wetting between the liquid filler metal and the surfaces of the cemented carbide and steel. The ultrasound generates cavitations within the liquid filler metal, which remove the oxides from the surface. Several filler metals such as a silver based alloy Ag449, pure Zn, and an AlSi-alloy were used to reduce the brazing temperature and to lower the thermal residual stresses within the joint. As a result, every filler metal successfully wetted both materials and led to a dense connection. The ultrasound has to be applied carefully to prevent a damage of the cemented carbide. In this regard, it was observed that single grains of the cemented carbide broke out and remained in the joint. This positive result of brazing cemented carbides to steel without a flux but using ultrasound, allows future studies to focus on the shear strength of these joints as well as the behavior of the thermally induced residual stresses.

  2. Endogenously Tagged Rab Proteins: A Resource to Study Membrane Trafficking in Drosophila

    PubMed Central

    Dunst, Sebastian; Kazimiers, Tom; von Zadow, Felix; Jambor, Helena; Sagner, Andreas; Brankatschk, Beate; Mahmoud, Ali; Spannl, Stephanie; Tomancak, Pavel; Eaton, Suzanne; Brankatschk, Marko

    2015-01-01

    Summary Membrane trafficking is key to the cell biological mechanisms underlying development. Rab GTPases control specific membrane compartments, from core secretory and endocytic machinery to less-well-understood compartments. We tagged all 27 Drosophila Rabs with YFPMYC at their endogenous chromosomal loci, determined their expression and subcellular localization in six tissues comprising 23 cell types, and provide this data in an annotated, searchable image database. We demonstrate the utility of these lines for controlled knockdown and show that similar subcellular localization can predict redundant functions. We exploit this comprehensive resource to ask whether a common Rab compartment architecture underlies epithelial polarity. Strikingly, no single arrangement of Rabs characterizes the five epithelia we examine. Rather, epithelia flexibly polarize Rab distribution, producing membrane trafficking architectures that are tissue- and stage-specific. Thus, the core machinery responsible for epithelial polarization is unlikely to rely on polarized positioning of specific Rab compartments. PMID:25942626

  3. Endogenously tagged rab proteins: a resource to study membrane trafficking in Drosophila.

    PubMed

    Dunst, Sebastian; Kazimiers, Tom; von Zadow, Felix; Jambor, Helena; Sagner, Andreas; Brankatschk, Beate; Mahmoud, Ali; Spannl, Stephanie; Tomancak, Pavel; Eaton, Suzanne; Brankatschk, Marko

    2015-05-04

    Membrane trafficking is key to the cell biological mechanisms underlying development. Rab GTPases control specific membrane compartments, from core secretory and endocytic machinery to less-well-understood compartments. We tagged all 27 Drosophila Rabs with YFP(MYC) at their endogenous chromosomal loci, determined their expression and subcellular localization in six tissues comprising 23 cell types, and provide this data in an annotated, searchable image database. We demonstrate the utility of these lines for controlled knockdown and show that similar subcellular localization can predict redundant functions. We exploit this comprehensive resource to ask whether a common Rab compartment architecture underlies epithelial polarity. Strikingly, no single arrangement of Rabs characterizes the five epithelia we examine. Rather, epithelia flexibly polarize Rab distribution, producing membrane trafficking architectures that are tissue- and stage-specific. Thus, the core machinery responsible for epithelial polarization is unlikely to rely on polarized positioning of specific Rab compartments.

  4. Molecular cloning, sequence characterization and expression pattern of Rab18 gene from watermelon (Citrullus lanatus).

    PubMed

    Xinli, Xiao; Lei, Peng

    2015-03-04

    The complete mRNA sequence of watermelon Rab18 gene was amplified through the rapid amplification of cDNA ends (RACE) method. The full-length mRNA was 1010 bp containing a 645 bp open reading frame, which encodes a protein of 214 amino acids. Sequence analysis revealed that watermelon Rab18 protein shares high homology with the Rab18 of cucumber (99%), muskmelon (98%), Morus notabilis (90%), tomato (89%), wine grape (89%) and potato (88%). Phylogenetic analysis revealed that watermelon Rab18 gene has a closer genetic relationship with Rab18 gene of cucumber and muskmelon. Tissue expression profile analysis indicated that watermelon Rab18 gene was highly expressed in root, stem and leaf, moderately expressed in flower and weakly expressed in fruit.

  5. A Rab3a-dependent complex essential for lysosome positioning and plasma membrane repair

    PubMed Central

    Encarnação, Marisa; Mateus, Denisa; Michelet, Xavier; Santarino, Inês; Hsu, Victor W.; Brenner, Michael B.

    2016-01-01

    Lysosome exocytosis plays a major role in resealing plasma membrane (PM) disruptions. This process involves two sequential steps. First, lysosomes are recruited to the periphery of the cell and then fuse with the damaged PM. However, the trafficking molecular machinery involved in lysosome exocytosis and PM repair (PMR) is poorly understood. We performed a systematic screen of the human Rab family to identify Rabs required for lysosome exocytosis and PMR. Rab3a, which partially localizes to peripheral lysosomes, was one of the most robust hits. Silencing of Rab3a or its effector, synaptotagmin-like protein 4a (Slp4-a), leads to the collapse of lysosomes to the perinuclear region and inhibition of PMR. Importantly, we have also identified a new Rab3 effector, nonmuscle myosin heavy chain IIA, as part of the complex formed by Rab3a and Slp4-a that is responsible for lysosome positioning at the cell periphery and lysosome exocytosis. PMID:27325790

  6. Ultrasonic guided wave inspection of Inconel 625 brazed lap joints

    NASA Astrophysics Data System (ADS)

    Comot, Pierre; Bocher, Philippe; Belanger, Pierre

    2016-04-01

    The aerospace industry has been investigating the use of brazing for structural joints, as a mean of reducing cost and weight. There therefore is a need for a rapid, robust, and cost-effective non-destructive testing method for evaluating the structural integrity of the joints. The mechanical strength of brazed joints depends mainly on the amount of brittle phases in their microstructure. Ultrasonic guided waves offer the possibility of detecting brittle phases in joints using spatio-temporal measurements. Moreover, they offer the opportunity to inspect complex shape joints. This study focused on the development of a technique based on ultrasonic guided waves for the inspection of Inconel 625 lap joints brazed with BNi-2 filler metal. A finite element model of a lap joint was used to optimize the inspection parameters and assess the feasibility of detecting the amount of brittle phases in the joint. A finite element parametric study simulating the input signal shape, the center frequency, and the excitation direction was performed. The simulations showed that the ultrasonic guided wave energy transmitted through, and reflected from, the joints was proportional to the amount of brittle phases in the joint.

  7. Mechanistic understanding of aerosol emissions from a brazing operation.

    PubMed

    Zimmer, A T; Biswas, P

    2000-01-01

    Welding operations produce gaseous and aerosol by-products that can have adverse health effects. A laboratory furnace study was conducted to aid understanding of the chemical and aerosol behavior of a widely used, self-fluxing brazing alloy (89% Cu, 6% Ag, 5% P) that is also used with a supplemental fluxing compound to prevent oxidation at the molten metal surface. The results indicate that the aerosols generated by the alloy are transient (produced over a short duration of time) and are associated with mass transfer of phosphorus species from the molten metal surface to the surrounding gas. In contrast, when the alloy was used in conjunction with the supplemental fluxing compound, a relatively nontransient, submicron-size aerosol was generated that was several orders of magnitude higher in concentration. Thermodynamic equilibrium analysis suggests that fluoride (a major constituent in the fluxing compound) played a significant role in reacting with the brazing alloy metals to form gas phase metal fluoride compounds that had high vapor pressures when compared with their elemental or oxide forms. As these metal-fluoride vapors cooled, submicron-size particles were formed mainly through nucleation and condensation growth processes. In addition, the equilibrium results revealed the potential formation of severe pulmonary irritants (HF and BF3) from heating the supplemental fluxing compound. These results demonstrated the importance of fluxing compounds in the formation of brazing fumes, and suggest that fluxing compounds could be selected that serve their metallurgical intention and suppress the formation of aerosols.

  8. Laser Brazing with Beam Scanning: Experimental and Simulative Analysis

    NASA Astrophysics Data System (ADS)

    Heitmanek, M.; Dobler, M.; Graudenz, M.; Perret, W.; Göbel, G.; Schmidt, M.; Beyer, E.

    Laser beam brazing with copper based filler wire is a widely established technology for joining zinc-coated steel plates in the body-shop. Successful applications are the divided tailgate or the zero-gap joint, which represents the joint between the side panel and the roof-top of the body-in-white. These joints are in direct view to the customer, and therefore have to fulfil highest optical quality requirements. For this reason a stable and efficient laser brazing process is essential. In this paper the current results on quality improvement due to one dimensional laser beam deflections in feed direction are presented. Additionally to the experimental results a transient three-dimensional simulation model for the laser beam brazing process is taken into account. With this model the influence of scanning parameters on filler wire temperature and melt pool characteristics is analyzed. The theoretical predictions are in good accordance with the experimental results. They show that the beam scanning approach is a very promising method to increase process stability and seam quality.

  9. Dispersed metal-toughened ceramics and ceramic brazing

    SciTech Connect

    Moorhead, A.J.; Tiegs, T.N.; Lauf, R.J.

    1983-01-01

    An alumina (Al/sub 2/O/sub 3/) based material that contains approximately 1 vol % finely dispersed platinum or chromium was developed for use in high temperature thermal-shock resistant electrical insulators. The work at ORNL is divided into two areas: (1) development of DMT ceramics; and (2) development of brazing filler metals suitable for making ceramic-to-ceramic and ceramic-to-metal brazements. The DMT ceramics and brazements are intended for service at elevated temperatures and at high stress levels in the dirty environments of advanced heat engines. The development and characterization of DMT ceramics includes processing (powder preparation, densification and heat treatment) and detailed measurement of mechanical and physical properties (strength, fracture toughness, and thermal conductivity). The brazing work includes: (1) the formulation and melting of small quantities of experimental brazing filler metals; (2) evaluation of the wetting and bonding behavior of these filler metals on Al/sub 2/O/sub 3/, partially stabilized zirconia and ..cap alpha..-SiC in a sessile drop apparatus; and (3) determine the short-term strength and fracture toughness of brazements.

  10. A study on brazing of Glidcop® to OFE Cu for application in Photon Absorbers of Indus-2

    NASA Astrophysics Data System (ADS)

    Yadav, D. P.; Kaul, R.; Ram Sankar, P.; Kak, A.; Ganesh, P.; Shiroman, R.; Singh, R.; Singh, A. P.; Tiwari, P.; Abhinandan, L.; Kukreja, L. M.; Shukla, S. K.

    2012-11-01

    The paper describes an experimental study aimed at standardizing brazing procedure for joining Glidcop to OFE Cu for its application in upgraded photon absorbers of 2.5 GeV synchrotron radiation source, Indus-2. Two different brazing routes, involving brazing with silver base (BVAg-8) and gold base (50Au/50Cu) alloys, were studied to join Glidcop to OFE Cu. Brazing with both alloys yielded helium leak tight and bakeable joints with acceptable shear strengths.

  11. Rab8 directs furrow ingression and membrane addition during epithelial formation in Drosophila melanogaster.

    PubMed

    Mavor, Lauren M; Miao, Hui; Zuo, Zhongyuan; Holly, Ryan M; Xie, Yi; Loerke, Dinah; Blankenship, J Todd

    2016-03-01

    One of the most fundamental changes in cell morphology is the ingression of a plasma membrane furrow. The Drosophila embryo undergoes several cycles of rapid furrow ingression during early development that culminate in the formation of an epithelial sheet. Previous studies have demonstrated the requirement for intracellular trafficking pathways in furrow ingression; however, the pathways that link compartmental behaviors with cortical furrow ingression events are unclear. Here, we show that Rab8 has striking dynamic behaviors in vivo. As furrows ingress, cytoplasmic Rab8 puncta are depleted and Rab8 accumulates at the plasma membrane in a location that coincides with known regions of directed membrane addition. We additionally use CRISPR/Cas9 technology to N-terminally tag Rab8, which is then used to address endogenous localization and function. Endogenous Rab8 displays partial coincidence with Rab11 and the Golgi, and this colocalization is enriched during the fast phase of cellularization. When Rab8 function is disrupted, furrow formation in the early embryo is completely abolished. We also demonstrate that Rab8 behaviors require the function of the exocyst complex subunit Sec5 as well as the recycling endosome protein Rab11. Active, GTP-locked Rab8 is primarily associated with dynamic membrane compartments and the plasma membrane, whereas GDP-locked Rab8 forms large cytoplasmic aggregates. These studies suggest a model in which active Rab8 populations direct furrow ingression by guiding the targeted delivery of cytoplasmic membrane stores to the cell surface through interactions with the exocyst tethering complex.

  12. Rab25 upregulation correlates with the proliferation, migration, and invasion of renal cell carcinoma

    SciTech Connect

    Li, Yuanyuan; Jia, Qingzhu; Zhang, Qian; Wan, Ying

    2015-03-20

    Renal cell carcinoma (RCC) is a common urological cancer with a poor prognosis. A recent cohort study revealed that the median survival of RCC patients was only 1.5 years and that <10% of the patients in the study survived up to 5 years. In tumor development, Rab GTPase are known to play potential roles such as regulation of cell proliferation, migration, invasion, communication, and drug resistance in multiple tumors. However, the correlation between Rabs expression and the occurrence, development, and metastasis of RCC remains unclear. In this study, we analyzed the transcriptional levels of 52 Rab GTPases in RCC patients. Our results showed that high levels of Rab25 expression were significantly correlated with RCC invasion classification (P < 0.01), lymph-node metastasis (P < 0.001), and pathological stage (P < 0.01). Conversely, in 786-O and A-498 cells, knocking down Rab25 protein expression inhibited cell proliferation, migration, and invasion. Our results also demonstrated that Rab25 is a target gene of let-7d, and further suggested that Rab25 upregulation in RCC is due to diminished expression of let-7d. These findings indicate that Rab25 might be a novel candidate molecule involved in RCC development, thus identifying a potential biological therapeutic target for RCC. - Highlights: • The transcriptional levels of 52 Rab GTPases were analyzed in renal cell carcinoma (RCC). • High levels of Rab25 expression were significantly correlated with clinicopathological factors of RCC. • Knockdown of Rab25 protein expression reduced RCC cells proliferation, migration, and invasion. • Rab25 is a target gene of let-7d in RCC.

  13. Insights into endosomal maturation of human holo-transferrin in the enteric parasite Entamoeba histolytica: essential roles of Rab7A and Rab5 in biogenesis of giant early endocytic vacuoles.

    PubMed

    Verma, Kuldeep; Saito-Nakano, Yumiko; Nozaki, Tomoyoshi; Datta, Sunando

    2015-12-01

    The pathogenic amoeba Entamoeba histolytica is one of the causative agents of health hazards in tropical countries. It causes amoebic dysentery, colitis and liver abscesses in human. Iron is one of the essential nutritional resources for survival and chronic infection caused by the amoeba. The parasite has developed multiple ways to import, sequester and utilize iron from various iron-binding proteins from its host. In spite of its central role in pathogenesis, the mechanism of iron uptake by the parasite is largely unknown. Here, we carried out a systematic study to understand the role of some of the amoebic homologues of mammalian endocytic Rab GTPases (Rab5 and Rab21, Rab7A and Rab7B) in intracellular transport of human holo-transferrin by the parasite. Flow cytometry and quantitative microscopic image analysis revealed that Rab5 and Rab7A are required for the biogenesis of amoebic giant endocytic vacuoles (GEVs) and regulate the early phase of intracellular trafficking of transferrin. Rab7B is involved in the late phase, leading to the degradation of transferrin in the amoebic lysosome-like compartments. Using time-lapse fluorescence imaging in fixed trophozoites, we determined the kinetics of the vesicular transport of transferrin through Rab5-, Rab7A- and Rab7B-positive compartments. The involvement of Rab7A in the early phase of endocytosis by the parasite marks a significant divergence from its host in terms of spatiotemporal regulation by the Rab GTPases.

  14. Critical importance of RAB proteins for synaptic function.

    PubMed

    Mignogna, Maria Lidia; D'Adamo, Patrizia

    2017-02-01

    Neurons are highly polarized cells that exhibit one of the more complex morphology and function. Neuronal intracellular trafficking plays a key role in dictating the directionality and specificity of vesicle formation, transport and fusion, allowing the transmission of information in sophisticate cellular network. Thus, the integrity of protein trafficking and spatial organization is especially important in neuronal cells. RAB proteins, small monomeric GTPases belonging to the RAS superfamily, spatially and temporally orchestrate specific vesicular trafficking steps. In this review we summarise the known roles of RAB GTPases involved in the maintenance of neuronal vesicular trafficking in the central nervous system. In particular, we discriminate the axonal pre-synaptic trafficking and dendritic post-synaptic trafficking, to better underlie how a correct orchestration of vesicle movement is necessary to maintain neuronal polarity and then, to permit an accurate architecture and functionality of synaptic activity.

  15. RAB1A promotes Vaccinia virus replication by facilitating the production of intracellular enveloped virions

    SciTech Connect

    Pechenick Jowers, Tali; Featherstone, Rebecca J.; Reynolds, Danielle K.; Brown, Helen K.; James, John; Prescott, Alan; Haga, Ismar R.; Beard, Philippa M.

    2015-01-15

    Vaccinia virus (VACV) is a large double-stranded DNA virus with a complex cytoplasmic replication cycle that exploits numerous cellular proteins. This work characterises the role of a proviral cellular protein, the small GTPase RAB1A, in VACV replication. Using siRNA, we identified RAB1A as required for the production of extracellular enveloped virions (EEVs), but not intracellular mature virions (IMVs). Immunofluorescence and electron microscopy further refined the role of RAB1A as facilitating the wrapping of IMVs to become intracellular enveloped virions (IEVs). This is consistent with the known function of RAB1A in maintenance of ER to Golgi transport. VACV can therefore be added to the growing list of viruses which require RAB1A for optimal replication, highlighting this protein as a broadly proviral host factor. - Highlights: • Characterisation of the role of the small GTPase RAB1A in VACV replication. • RAB1A is not required for production of the primary virion form (IMV). • RAB1A is required for production of processed virion forms (IEVs, CEVs and EEVs). • Consistent with known role of RAB1A in ER to Golgi transport.

  16. Drosophila Rab14 mediates phagocytosis in the immune response to Staphylococcus aureus

    PubMed Central

    Garg, Aprajita; Wu, Louisa P.

    2014-01-01

    Drosophila hemocytes are essential for the animal to resist Staphylococcus aureus infections. Phagocytosis is a central component of the hemocyte-mediated immune response. It involves regulated interaction between the phagocytic and the endocytic compartments. Rab GTPases are pivotal for the membrane trafficking and fusion events, and thus are often targets of intracellular pathogens that subvert phagocytosis. An in vivo screen identified Rab2 and Rab14 as candidates for proteins regulating phagosome maturation. Since Rab14 is often targeted by intracellular pathogens, an understanding of its function during phagocytosis and the overall immune response can give insight into the pathogenesis of intracellular microbes. We generated a Drosophila Rab14 mutant and characterized the resulting immune defects in animals and specifically in hemocytes in response to an S. aureus infection. Hemocyte based immunofluorescence studies indicate that Rab14 is recruited to the phagosome and like Rab7, a well-characterized regulator of the phagocytic pathway, is essential for progression of phagosome maturation. Rab14 mutant hemocytes show impaired recruitment of Rab7 and of a lysosomal marker onto S. aureus phagosomes. The defect in phagocytosis is associated with higher bacterial load and increased susceptibility to S. aureus in the animal. PMID:24119134

  17. Association of the GTP-binding protein Rab3A with bovine adrenal chromaffin granules

    SciTech Connect

    Darchen, F.; Hammel, F.; Monteils, M.P.; Scherman, D. ); Zahraoui, A.; Tavitian, A. )

    1990-08-01

    The Rab3A protein belongs to a large family of small GTP-binding proteins that are present in eukaryotic cells and that share amino acid identities with the Ras proteins (products of the ras protooncogenes). Rab3A, which is specifically located in nervous and endocrine tissues, is suspected to play a key role in secretion. Its localization was investigated in bovine adrenal gland by using a polyclonal antibody. Rab3A was detected in adrenal medulla but not in adrenal cortex. In cultured adrenal medulla cells, Rab3A was specifically expressed in the catecholamine-secreting chromaffin cells. Subcellular fractionation suggested that Rab3A is about 30% cytosolic and that particulate Rab3A is associated with the membrane of chromaffin granules (the catecholamine storage organelles) and with a second compartment likely to be the plasma membrane. The Rab3A localization on chromaffin granule membranes was confirmed by immunoadsorption with an antibody against dopamine {beta}-hydroxylase. Rab3A was not extracted from this membrane by NaCl or KBr but was partially extracted by urea and totally solubilized by Triton X-100, suggesting either an interaction with an intrinsic protein or a membrane association through fatty acid acylation. This study suggests that Rab3A, which may also be located on other secretory vesicles containing noncharacterized small GTP-binding proteins, is involved in their biogenesis or in the regulated secretion process.

  18. Down-regulation of Rab5 decreases characteristics associated with maintenance of cell transformation

    SciTech Connect

    Silva, Patricio; Soto, Nicolás; Díaz, Jorge; Mendoza, Pablo; Díaz, Natalia; Quest, Andrew F.G.; Torres, Vicente A.

    2015-08-21

    The early endosomal protein Rab5 is highly expressed in tumor samples, although a causal relationship between Rab5 expression and cell transformation has not been established. Here, we report the functional effects of targeting endogenous Rab5 with specific shRNA sequences in different tumor cell lines. Rab5 down-regulation in B16-F10 cells decreased tumor formation by subcutaneous injection into C57/BL6 mice. Accordingly, Rab5 targeting in B16-F10 and A549, but not MDA-MB-231 cells was followed by decreased cell proliferation, increased apoptosis and decreased anchorage-independent growth. These findings suggest that Rab5 expression is required to maintain characteristics associated with cell transformation. - Highlights: • Rab5 is important to the maintenance of cell transformation characteristics. • Down-regulation of Rab5 decreases cell proliferation and increases apoptosis in different cancer cells. • Rab5 is required for anchorage-independent growth and tumorigenicity in-vivo.

  19. Rab14 Act as Oncogene and Induce Proliferation of Gastric Cancer Cells via AKT Signaling Pathway

    PubMed Central

    Zhao, Zhenghao; Li, Qian; Zhou, Kaiyue; Zhao, Lingyu; Wang, Lumin; Yang, Juan; Huang, Chen

    2017-01-01

    Rab14 is a member of RAS oncogene family, and its dysfunction has been reported to be involved in various types of human cancer. However, its expression and function were still unclear in gastric cancer. The aim of this study was to investigate the function and mechanism of Rab14 in gastric cancer cell lines. Quantitative real-time PCR (qRT-PCR) was performed in 17 gastric adenocarcinoma tissues and 4 cell lines to detect the expression of Rab14. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT), colony formation and flow cytometry assays were employed to determine the proliferative ability, cell cycle transition and apoptosis in vitro in BGC-823 or SGC-7901 cells. Western blot was performed to investigate the pathways and mechanism of Rab14 regulation. In this study, we show that Rab14 presents a significant up-regulated expression among the paired tissue samples and cell lines in gastric cancer. When we overexpressed Rab14 in SGC-7901 cells or silenced Rab14 in BGC-823 cells, we found that Rab14 could modify cell growth, cell cycle or apoptosis, which accompanied with an obvious regulation of CCND1, CDK2 and BAX involving in AKT signaling pathway. In conclusion, this study provides a new evidence on that Rab14 functions as a novel tumor oncogene and could be a potential therapeutic target in gastric cancer. PMID:28107526

  20. Increased Rab35 expression is a potential biomarker and implicated in the pathogenesis of Parkinson's disease

    PubMed Central

    Lai, Szu-Chia; Weng, Yi-Hsin; Huang, Yin-Cheng; Cheng, Yi-Chuan; Chen, Rou-Shayn; Huang, Ying-Zu; Hung, June; Chen, Chiung-Chu; Lin, Wey-Yil; Chang, Hsiu-Chen; Chen, Yu-Jie; Chen, Chao-Lang; Chen, Hsin-Yi; Lin, Yan-Wei; Wu-Chou, Yah-Huei; Wang, Hung-Li; Lu, Chin-Song

    2016-01-01

    Parkinson's disease (PD) is the second common neurodegenerative disease. Identification of biomarkers for early diagnosis and prediction of disease progression is important. The present comparative proteomic study of serum samples using two-dimensional fluorescence differential gel electrophoresis followed by ELISA confirmation demonstrated that protein expression of Rab35 was increased in PD patients compared with matched control subjects and other parkinsonian disorders, progressive supranuclear palsy (PSP) and multiple system atrophy (MSA). The serum level of Rab35 was significantly correlated with the age at onset of PD. The median age of onset in patients with higher Rab35 serum level was 5 years younger than those with lower Rab35 serum level. There was a positive correlation between the Rab35 level and disease duration of PD. Moreover, the protein expression of Rab35 was increased in the substantia nigra but not in the striatum of mouse models of PD, including MPTP-treated mice, rotenone-treated mice, (R1441C) LRRK2 or (G2019S) LRRK2 transgenic mice. Furthermore, overexpression of Rab35 increased the aggregation and secretion of mutant A53T α-synuclein in dopaminergic SH-SY5Y cells. Co-expression of Rab35 with wild-type or A53T α-synuclein in SH-SY5Y cells deteriorated cell death. Our results suggest that Rab35 is potentially useful in the differential diagnosis of parkinsonian disorders and is implicated in the pathogenesis of PD. PMID:27509057

  1. Rab from the white shrimp Litopenaeus vannamei: characterization and its regulation upon environmental stress.

    PubMed

    Wang, Lei; Wang, Xiao-Rong; Liu, Jin; Chen, Chu-Xian; Liu, Yuan; Wang, Wei-Na

    2015-10-01

    With the destruction of the ecological environment, shrimp cultivation in China has been seriously affected by outbreaks of infectious diseases. Rab, which belong to small GTPase Ras superfamily, can regulate multiple steps in eukaryotic vesicle trafficking including vesicle budding, vesicle tethering, and membrane fusion. Knowledge of Rab in shrimp is essential to understanding regulation and detoxification mechanisms of environmental stress. In this study, we analyzed the functions of Rab from the Pacific white shrimp, Litopenaeus vannamei. Full-length cDNA of Rab was obtained, which was 751 bp long, with open reading frame encoding 206 amino acids. In this study, for the first time, the gene expression of Rab of L. vannamei was analyzed by quantitative real-time PCR after exposure to five kinds of environmental stresses (bacteria, pH, Cd, salinity and low temperature). The results demonstrate that Rab is sensitive and involved in bacteria, pH, and Cd stress responses and Rab is more sensitive to bacteria than other stresses. Therefore we infer that Rab may have relationship with the anti-stress mechanism induced by environment stress in shrimp and Rab could be used as critical biomarkers for environmental quality assessment.

  2. The small GTPase Rab29 is a common regulator of immune synapse assembly and ciliogenesis

    PubMed Central

    Onnis, A; Finetti, F; Patrussi, L; Gottardo, M; Cassioli, C; Spanò, S; Baldari, C T

    2015-01-01

    Accumulating evidence underscores the T-cell immune synapse (IS) as a site of intense vesicular trafficking, on which productive signaling and cell activation crucially depend. Although the T-cell antigen receptor (TCR) is known to exploit recycling to accumulate to the IS, the specific pathway that controls this process remains to be elucidated. Here we demonstrate that the small GTPase Rab29 is centrally implicated in TCR trafficking and IS assembly. Rab29 colocalized and interacted with Rab8, Rab11 and IFT20, a component of the intraflagellar transport system that regulates ciliogenesis and participates in TCR recycling in the non-ciliated T cell, as assessed by co-immunoprecipitation and immunofluorescence analysis. Rab29 depletion resulted in the inability of TCRs to undergo recycling to the IS, thereby compromizing IS assembly. Under these conditions, recycling TCRs accumulated in Rab11+ endosomes that failed to polarize to the IS due to defective Rab29-dependent recruitment of the dynein microtubule motor. Remarkably, Rab29 participates in a similar pathway in ciliated cells to promote primary cilium growth and ciliary localization of Smoothened. These results provide a function for Rab29 as a regulator of receptor recycling and identify this GTPase as a shared participant in IS and primary cilium assembly. PMID:26021297

  3. Rab7b at the intersection of intracellular trafficking and cell migration

    PubMed Central

    Distefano, Marita Borg; Kjos, Ingrid; Bakke, Oddmund; Progida, Cinzia

    2015-01-01

    Abstract Rab proteins are small GTPases essential for controlling and coordinating intracellular traffic. The small GTPase Rab7b regulates the retrograde transport from late endosomes toward the Trans-Golgi Network (TGN), and is important for the proper trafficking of several receptors such as Toll-like receptors (TLRs) and sorting receptors. We recently identified the actin motor protein myosin II as a new interaction partner for Rab7b, and found that Rab7b transport is dependent on myosin II. Interestingly, we also discovered that Rab7b influences the phosphorylation state of myosin II by controlling the activation status of the small GTPase RhoA. Consequently, Rab7b is important for the remodeling of actin filaments in processes such as stress fiber formation, cell adhesion, polarization and cell migration. Our finding that Rab7b can control actomyosin reorganization reveals yet another important role for Rab proteins, in addition to their already established role as master regulators of intracellular transport. Here we discuss our findings and speculate how they can explain the importance of Rab7b in dendritic cells (DCs). PMID:27066171

  4. Rab7b at the intersection of intracellular trafficking and cell migration.

    PubMed

    Distefano, Marita Borg; Kjos, Ingrid; Bakke, Oddmund; Progida, Cinzia

    2015-01-01

    Rab proteins are small GTPases essential for controlling and coordinating intracellular traffic. The small GTPase Rab7b regulates the retrograde transport from late endosomes toward the Trans-Golgi Network (TGN), and is important for the proper trafficking of several receptors such as Toll-like receptors (TLRs) and sorting receptors. We recently identified the actin motor protein myosin II as a new interaction partner for Rab7b, and found that Rab7b transport is dependent on myosin II. Interestingly, we also discovered that Rab7b influences the phosphorylation state of myosin II by controlling the activation status of the small GTPase RhoA. Consequently, Rab7b is important for the remodeling of actin filaments in processes such as stress fiber formation, cell adhesion, polarization and cell migration. Our finding that Rab7b can control actomyosin reorganization reveals yet another important role for Rab proteins, in addition to their already established role as master regulators of intracellular transport. Here we discuss our findings and speculate how they can explain the importance of Rab7b in dendritic cells (DCs).

  5. Recent experience in the fabrication and brazing of ceramic beam tubes for kicker magnets at FNAL

    SciTech Connect

    Ader, C.R.; Jensen, C.; Reilly, R.; Snee, D.; Wilson, J.H.; /Fermilab

    2008-06-01

    Ceramic beam tubes are utilized in numerous kicker magnets in different accelerator rings at Fermi National Accelerator Laboratory. Kovar flanges are brazed onto each beam tube end, since kovar and high alumina ceramic have similar expansion curves. The tube, kovar flange, end piece, and braze foil (titanium/incusil) alloy brazing material are stacked in the furnace and then brazed in the furnace at 1000 C. The ceramic specified is 99.8% Alumina, Al{sub 2}O{sub 3}, a strong recrystallized high-alumina fabricated by slip casting. Recent experience at Fermilab with the fabrication and brazing of these tubes has brought to light numerous problems including tube breakage and cracking and also the difficulty of brazing the tube to produce a leak-tight joint. These problems may be due to the ceramic quality, voids in the ceramic, thinness of the wall, and micro-cracks in the ends which make it difficult to braze because it cannot fill tiny surface cracks which are caused by grain pullout during the cutting process. Solutions which are being investigated include lapping the ends of the tubes before brazing to eliminate the micro-cracks and also metallization of the tubes.

  6. Wettability of brazing alloys on molybdenum and TZM (Mo-Ti-Zr alloy)

    SciTech Connect

    McDonald, M.M.; Keller, D.L.; Heiple, C.R.; Hofmann, W.E.

    1988-01-01

    Vacuum brazing studies have been performed on molybdenum and TZM (0.5Ti-0.08Zr-Mo). Wettability tests have been conducted for nineteen braze metal filler alloys on molybdenum and thirty-two braze metal filler alloys on TZM over a wide range of temperatures. A wetting index, which is a function of contact angle and braze alloy contact area, was determined for each filler alloy at each brazing temperature. The nature and extent of interaction between the brazing alloys and the base metals was analyzed by conventional metallography, scanning-electron microscopy, and electron microprobe analysis. A comparison is made between the behavior of filler alloys on molybdenum and TZM -- filler alloys consistently exhibited less wettability on TZM than on molybdenum. The lower wettability of TZM is believed to be due to a small amount of titanium in the surface oxide on TZM. Cracking was observed in the base metal under some of the high temperature braze deposits. The cracking is shown to arise from liquid metal embrittlement from nickel in the high temperature braze alloys. 7 refs., 11 figs., 2 tabs.

  7. Strength and structure of furnace-brazed joints between aluminum and stainless steel

    SciTech Connect

    Roulin, M.; Karadeniz, G.; Mortensen, A.; Luster, J.W.

    1999-05-01

    The structure and shear strength of brazed joints of aluminum to stainless steel are studied using a modification of the double lap joint configuration, which allows mechanical testing and joint microstructure examination on the same test piece. It is found that during furnace brazing of such joints at 600 C, using an Al-Si eutectic brazing alloy, the interfacial zone between the aluminum-rich braze and the stainless steel substrate features two intermetallic layers. The first is formed in the initial instants of the process and features an overall composition similar to that of the compound FeSiAl{sub 5}. The second appears after a 10-min hold time at the brazing temperature, and features an overall composition that parallels the FeAl{sub 3} intermetallic. Both layers are, however, more complex in structure than is suggested by these stoichiometric relations. The shear strength of the braze peaks at 21 MPa after a 10-min hold time at the brazing temperature. This peak is associated with nucleation of the second intermetallic layer, which is shown to fragilize the joint significantly. The presence of silicon in the brazing alloy would also seem to be beneficial by retarding formation of this second, more fragile Fe-Al intermetallic layer; however, more work is needed to substantiate this tentative conclusion.

  8. Interfacial metallurgy study of brazed joints between tungsten and fusion related materials for divertor design

    NASA Astrophysics Data System (ADS)

    Zhang, Yuxuan; Galloway, Alexander; Wood, James; Robbie, Mikael Brian Olsson; Easton, David; Zhu, Wenzhong

    2014-11-01

    In the developing DEMO divertor, the design of joints between tungsten to other fusion related materials is a significant challenge as a result of the dissimilar physical metallurgy of the materials to be joined. This paper focuses on the design and fabrication of dissimilar brazed joints between tungsten and fusion relevant materials such as EUROFER 97, oxygen-free high thermal conductivity (OFHC) Cu and SS316L using a gold based brazing foil. The main objectives are to develop acceptable brazing procedures for dissimilar joining of tungsten to other fusion compliant materials and to advance the metallurgical understanding within the interfacial region of the brazed joint. Four different butt-type brazed joints were created and characterised, each of which were joined with the aid of a thin brazing foil (Au80Cu19Fe1, in wt.%). Microstructural characterisation and elemental mapping in the transition region of the joint was undertaken and, thereafter, the results were analysed as was the interfacial diffusion characteristics of each material combination produced. Nano-indentation tests are performed at the joint regions and correlated with element composition information in order to understand the effects of diffused elements on mechanical properties. The experimental procedures of specimen fabrication and material characterisation methods are presented. The results of elemental transitions after brazing are reported. Elastic modulus and nano-hardness of each brazed joints are reported.

  9. Substrate Effects on the High Temperature Oxidation Behavior of a Gold-Based Braze Filler Metal

    SciTech Connect

    Weil, K. Scott; Rice, Joseph P.

    2005-06-01

    Oxidation testing was conducted on a commercial gold-based braze alloy, Gold ABA®, and on zirconia/stainless steel couples joined using this filler metal. Preliminary results reveal that both substrates play a significant role in determining the overall oxidation behavior of the brazed joint.

  10. Reduction of liquid metal embrittlement in copper-brazed stainless steel joints

    NASA Astrophysics Data System (ADS)

    Uhlig, T.; Fedorov, V.; Elßner, M.; Wagner, G.; Weis, S.

    2017-03-01

    Due to its very good formability and the low raw material cost, pure copper in form of foils is commonly used to braze plate heat exchangers made of stainless steel. The difference in the electrochemical potentials of brazing filler and base material leads to corrosion effects in contact with electrolytes. This may lead to leakages, which decrease the reliability of the heat exchanger during service in potable water. The dissolution of the emerging corrosion products of brazing filler and base material induces the migration of heavy metal ions, such as Cu2+ and Ni2+, into the potable water. The so-called liquid metal embrittlement, which takes place during the brazing process, may intensify the corrosion. The brazing filler infiltrates the stainless steel along the grain boundaries and causes an embrittlement. This paper deals with the determination of the grain boundary erosion dependent on the degree of deformation and heat treatment of the stainless steel AISI 316L.

  11. Ultrasonic C-scan evaluation of Ti-alloy/Al brazed joint

    NASA Astrophysics Data System (ADS)

    Murthy, G. V. S.; Sridhar, G.

    2013-01-01

    An indigenously developed brazed joint of Al/Ti was characterized by ultrasonic C-scan method. The study revealed certain debond locations in the brazed joint. These debond locations would have resulted due to the failure of filler material's joining ability. In spite of certain amount of load applied on the braze base metals, the above debonding happened. The debond locations further confirm that the pressure was not sufficient and therefore the debonds are present at exactly the same location where there were gaps in the fixture. Further analysis revealed that these debonds were present at the interface between the braze material and Al-plate. It is concluded that ultrasonic C-scan can be effectively used to evaluate the bond characteristics in a brazed joint between dissimilar metals.

  12. Hypoxia promotes Rab5 activation, leading to tumor cell migration, invasion and metastasis.

    PubMed

    Silva, Patricio; Mendoza, Pablo; Rivas, Solange; Díaz, Jorge; Moraga, Carolina; Quest, Andrew F G; Torres, Vicente A

    2016-05-17

    Hypoxia, a common condition of the tumor microenvironment, is associated with poor patient prognosis, tumor cell migration, invasion and metastasis. Recent evidence suggests that hypoxia alters endosome dynamics in tumor cells, leading to augmented cell proliferation and migration and this is particularly relevant, because endosomal components have been shown to be deregulated in cancer. The early endosome protein Rab5 is a small GTPase that promotes integrin trafficking, focal adhesion turnover, Rac1 activation, tumor cell migration and invasion. However, the role of Rab5 and downstream events in hypoxia remain unknown. Here, we identify Rab5 as a critical player in hypoxia-driven tumor cell migration, invasion and metastasis. Exposure of A549 human lung carcinoma, ZR-75, MDA-MB-231 and MCF-7 human breast cancer and B16-F10 mouse melanoma cells to hypoxia increased Rab5 activation, followed by its re-localization to the leading edge and association with focal adhesions. Importantly, Rab5 was required for hypoxia-driven cell migration, FAK phosphorylation and Rac1 activation, as shown by shRNA-targeting and transfection assays with Rab5 mutants. Intriguingly, the effect of hypoxia on both Rab5 activity and migration was substantially higher in metastatic B16-F10 cells than in poorly invasive B16-F0 cells. Furthermore, exogenous expression of Rab5 in B16-F0 cells predisposed to hypoxia-induced migration, whereas expression of the inactive mutant Rab5/S34N prevented the migration of B16-F10 cells induced by hypoxia. Finally, using an in vivo syngenic C57BL/6 mouse model, Rab5 expression was shown to be required for hypoxia-induced metastasis. In summary, these findings identify Rab5 as a key mediator of hypoxia-induced tumor cell migration, invasion and metastasis.

  13. High Rab11-FIP4 expression predicts poor prognosis and exhibits tumor promotion in pancreatic cancer

    PubMed Central

    He, Yun; Ye, Mengsi; Zhou, Lingling; Shan, Yunfeng; Lu, Guangrong; Zhou, Yuhui; Zhong, Jinwei; Zheng, Jihang; Xue, Zhanxiong; Cai, Zhenzhai

    2017-01-01

    Some studies have demonstrated that Rab11-family interacting proteins (Rab11-FIPs) are connected with the tumorigenesis, and they may act as tumor promoters in some cancers. The clinicopathological significance of Rab11-family interacting protein 4 (Rab11-FIP4) expression and its possible effects on pancreatic cancer (PC) are still undiscovered. In this study, Rab11-FIP4 protein expression level in 60 PC specimens and pair-matched non-cancerous samples were detected by immunohistochemistry analysis. The results were analysed and compared with each patients' clinical data. Rab11-FIP4 expression in PC tissues increased significantly more than that of adjacent non-cancerous tissues (P=0.0001). Overexpression of Rab11-FIP4 in the PC tissues was significantly related to tumor size (P=0.0001), histological grade (P=0.028), metastasis (P=0.001) and TNM stage (P=0.004) but not with age (P=0.832), gender (P=0.228) or tumor site (P=0.875). Kaplan-Meier survival analysis showed that overexpression of Rab11-FIP4 was significantly related to overall survival time (P=0.0036). In addition, Rab11-FIP4 in PANC-1 pancreatic cancer cells were successfully knocked-out using the CRISPR/Cas9 system. Rab11-FIP4 knockout in PANC-1 cells inhibited cell growth, invasion and metastasis, and arrested cell cycle progression, but did not alter apoptosis. Our findings suggest that overexpression of Rab11-FIP4 predicts poor clinical outcomes for pancreatic cancer and contributes to pancreatic tumor progression. PMID:28035375

  14. Hypoxia promotes Rab5 activation, leading to tumor cell migration, invasion and metastasis

    PubMed Central

    Silva, Patricio; Mendoza, Pablo; Rivas, Solange; Díaz, Jorge; Moraga, Carolina; Quest, Andrew F.G.; Torres, Vicente A.

    2016-01-01

    Hypoxia, a common condition of the tumor microenvironment, is associated with poor patient prognosis, tumor cell migration, invasion and metastasis. Recent evidence suggests that hypoxia alters endosome dynamics in tumor cells, leading to augmented cell proliferation and migration and this is particularly relevant, because endosomal components have been shown to be deregulated in cancer. The early endosome protein Rab5 is a small GTPase that promotes integrin trafficking, focal adhesion turnover, Rac1 activation, tumor cell migration and invasion. However, the role of Rab5 and downstream events in hypoxia remain unknown. Here, we identify Rab5 as a critical player in hypoxia-driven tumor cell migration, invasion and metastasis. Exposure of A549 human lung carcinoma, ZR-75, MDA-MB-231 and MCF-7 human breast cancer and B16-F10 mouse melanoma cells to hypoxia increased Rab5 activation, followed by its re-localization to the leading edge and association with focal adhesions. Importantly, Rab5 was required for hypoxia-driven cell migration, FAK phosphorylation and Rac1 activation, as shown by shRNA-targeting and transfection assays with Rab5 mutants. Intriguingly, the effect of hypoxia on both Rab5 activity and migration was substantially higher in metastatic B16-F10 cells than in poorly invasive B16-F0 cells. Furthermore, exogenous expression of Rab5 in B16-F0 cells predisposed to hypoxia-induced migration, whereas expression of the inactive mutant Rab5/S34N prevented the migration of B16-F10 cells induced by hypoxia. Finally, using an in vivo syngenic C57BL/6 mouse model, Rab5 expression was shown to be required for hypoxia-induced metastasis. In summary, these findings identify Rab5 as a key mediator of hypoxia-induced tumor cell migration, invasion and metastasis. PMID:27121131

  15. Rab4GTPase modulates CFTR function by impairing channel expression at plasma membrane

    SciTech Connect

    Saxena, Sunil K. . E-mail: ssaxena@stevens.edu; Kaur, Simarna; George, Constantine

    2006-03-03

    Cystic fibrosis (CF), an autosomal recessive disorder, is caused by the disruption of biosynthesis or the function of a membrane cAMP-activated chloride channel, CFTR. CFTR regulatory mechanisms include recruitment of channel proteins to the cell surface from intracellular pools and by protein-protein interactions. Rab proteins are small GTPases involved in regulated trafficking controlling vesicle docking and fusion. Rab4 controls recycling events from endosome to the plasma membrane, fusion, and degradation. The colorectal cell line HT-29 natively expresses CFTR and responds to cAMP stimulation with an increase in CFTR-mediated currents. Rab4 over-expression in HT-29 cells inhibits both basal and cAMP-stimulated CFTR-mediated currents. GTPase-deficient Rab4Q67L and GDP locked Rab4S22N both inhibit channel activity, which appears characteristically different. Active status of Rab4 was confirmed by GTP overlay assay, while its expression was verified by Western blotting. The pull-down and immunoprecipitation experiments suggest that Rab4 physically interacts with CFTR through protein-protein interaction. Biotinylation with cell impermeant NHS-Sulfo-SS-Biotin implies that Rab4 impairs CFTR expression at cell surface. The enhanced cytosolic CFTR indicates that Rab4 expression restrains CFTR appearance at the cell membrane. The study suggests that Rab4 regulates the channel through multiple mechanisms that include protein-protein interaction, GTP/GDP exchange, and channel protein trafficking. We propose that Rab4 is a dynamic molecule with a significant role in CFTR function.

  16. Subcellular Localization and Functional Analysis of the Arabidopsis GTPase RabE1[W][OA

    PubMed Central

    Speth, Elena Bray; Imboden, Lori; Hauck, Paula; He, Sheng Yang

    2009-01-01

    Membrane trafficking plays a fundamental role in eukaryotic cell biology. Of the numerous known or predicted protein components of the plant cell trafficking system, only a relatively small subset have been characterized with respect to their biological roles in plant growth, development, and response to stresses. In this study, we investigated the subcellular localization and function of an Arabidopsis (Arabidopsis thaliana) small GTPase belonging to the RabE family. RabE proteins are phylogenetically related to well-characterized regulators of polarized vesicle transport from the Golgi apparatus to the plasma membrane in animal and yeast cells. The RabE family of GTPases has also been proposed to be a putative host target of AvrPto, an effector protein produced by the plant pathogen Pseudomonas syringae, based on yeast two-hybrid analysis. We generated transgenic Arabidopsis plants that constitutively expressed one of the five RabE proteins (RabE1d) fused to green fluorescent protein (GFP). GFP-RabE1d and endogenous RabE proteins were found to be associated with the Golgi apparatus and the plasma membrane in Arabidopsis leaf cells. RabE down-regulation, due to cosuppression in transgenic plants, resulted in drastically altered leaf morphology and reduced plant size, providing experimental evidence for an important role of RabE GTPases in regulating plant growth. RabE down-regulation did not affect plant susceptibility to pathogenic P. syringae bacteria; conversely, expression of the constitutively active RabE1d-Q74L enhanced plant defenses, conferring resistance to P. syringae infection. PMID:19233904

  17. Synaptic Function of Rab11Fip5: Selective Requirement for Hippocampal Long-Term Depression

    PubMed Central

    Ahmad, Mohiuddin; Jurado, Sandra; Malenka, Robert C.

    2015-01-01

    Postsynaptic AMPA-type glutamate receptors (AMPARs) are among the major determinants of synaptic strength and can be trafficked into and out of synapses. Neuronal activity regulates AMPAR trafficking during synaptic plasticity to induce long-term changes in synaptic strength, including long-term potentiation (LTP) and long-term depression (LTD). Rab family GTPases regulate most membrane trafficking in eukaryotic cells; particularly, Rab11 and its effectors are implicated in mediating postsynaptic AMPAR insertion during LTP. To explore the synaptic function of Rab11Fip5, a neuronal Rab11 effector and a candidate autism-spectrum disorder gene, we performed shRNA-mediated knock-down and genetic knock-out (KO) studies. Surprisingly, we observed robust shRNA-induced synaptic phenotypes that were rescued by a Rab11Fip5 cDNA but that were nevertheless not observed in conditional KO neurons. Both in cultured neurons and acute slices, KO of Rab11Fip5 had no significant effect on basic parameters of synaptic transmission, indicating that Rab11Fip5 is not required for fundamental synaptic operations, such as neurotransmitter release or postsynaptic AMPAR insertion. KO of Rab11Fip5 did, however, abolish hippocampal LTD as measured both in acute slices or using a chemical LTD protocol in cultured neurons but did not affect hippocampal LTP. The Rab11Fip5 KO mice performed normally in several behavioral tasks, including fear conditioning, but showed enhanced contextual fear extinction. These are the first findings to suggest a requirement for Rab11Fip5, and presumably Rab11, during LTD. PMID:25972173

  18. Interfacial segregation of Ti in the brazing of diamond grits onto a steel substrate using a Cu-Sn-Ti brazing alloy

    NASA Astrophysics Data System (ADS)

    Li, Wen-Chung; Lin, Shun-Tian; Liang, Cheng

    2002-07-01

    Diamond grits were brazed onto a steel substrate using a prealloyed Cu-10Sn-15Ti (wt pct) brazing alloy at 925 °C and 1050 °C. Due to the relatively high concentration of Ti in the brazing alloy, the braze matrix exhibited a composite structure, composed of β-(Cu,Sn), a Cu-based solid solution, and various intermetallic compounds with different morphologies. The reaction of Ti with diamond yielded a continuous TiC layer on the surfaces of the diamond grits. On top of the TiC growth front, an intermetallic compound, composed of Sn and Ti, nucleated and grew into a randomly interwoven fine lacey structure. An interfacial structure developed as the interwoven fine lacey phase was semicoherently bonded to the TiC layer, with the Cu-based braze matrix filling its interstices. The thickness of such a composite layer was increased linearly with the square root of isothermal holding time at 925 °C, complying with the law of a diffusion-controlled process. However, at 1050 °C, the segregation behavior of Ti and Sn to the interfaces between the TiC layer and the braze matrix diminished, due to the increased solubility of Ti in the Cu-based liquid phase. The enhanced dissolution of Ti in the Cu-based liquid phase at 1050 °C also caused the precipitation of rod-like CuTi with an average diameter of about 0.2 µm during cooling. SnTi3 was the predominant intermetallic compound and existed in three different forms in the braze matrix. It existed as interconnected grains of large size which either floated to the surface of the braze matrix or grew into faceted grains. It also exhibited a nail-like structure with a mean diameter of about 1 µm for the rod section and a lamellar structure arising from a eutectic reaction during cooling.

  19. Rab7 Regulates CDH1 Endocytosis, Circular Dorsal Ruffles Genesis, and Thyroglobulin Internalization in a Thyroid Cell Line.

    PubMed

    Mascia, Anna; Gentile, Flaviana; Izzo, Antonella; Mollo, Nunzia; De Luca, Maria; Bucci, Cecilia; Nitsch, Lucio; Calì, Gaetano

    2016-08-01

    Rab7 regulates the biogenesis of late endosomes, lysosomes, and autophagosomes. It has been proposed that a functional and physical interaction exists between Rab7 and Rac1 GTPases in CDH1 endocytosis and ruffled border formation. In FRT cells over-expressing Rab7, increased expression and activity of Rac1 was observed, whereas a reduction of Rab7 expression by RNAi resulted in reduced Rac1 activity, as measured by PAK1 phosphorylation. We found that CDH1 endocytosis was extremely reduced only in Rab7 over-expressing cells but was unchanged in Rab7 silenced cells. In Rab7 under or over-expressing cells, Rab7 and LC3B-II co-localized and co-localization in large circular structures occurred only in Rab7 over-expressing cells. These large circular structures occurred in about 10% of the cell population; some of them (61%) showed co-localization of Rab7 with cortactin and f-actin and were identified as circular dorsal ruffles (CDRs), the others as mature autophagosomes. We propose that the over-expression of Rab7 is sufficient to induce CDRs. Furthermore, in FRT cells, we found that the expression of the insoluble/active form of Rab7, rather than Rab5, or Rab8, was inducible by cAMP and that cAMP-stimulated FRT cells showed increased PAK1 phosphorylation and were no longer able to endocytose CDH1. Finally, we demonstrated that Rab7 over-expressing cells are able to endocytose exogenous thyroglobulin via pinocytosis/CDRs more efficiently than control cells. We propose that the major thyroglobulin endocytosis described in thyroid autonomous adenomas due to Rab7 increased expression, occurs via CDRs. J. Cell. Physiol. 231: 1695-1708, 2016. © 2015 Wiley Periodicals, Inc.

  20. Marked amplification and diversification of products of ras genes from rat brain, Rab GTPases, in the ciliates Tetrahymena thermophila and Paramecium tetraurelia.

    PubMed

    Saito-Nakano, Yumiko; Nakahara, Tohru; Nakano, Kentaro; Nozaki, Tomoyoshi; Numata, Osamu

    2010-01-01

    Small GTPase Rab (products of ras genes from rat brain) is a widely conserved molecular switch among eukaryotes and regulates membrane trafficking pathways. It is generally considered that the number of Rab encoded in the genome correlates with multicellularity; however, we found that unicellular ciliates Tetrahymena thermophila (Tt) and Paramecium tetraurelia (Pt) possess many more Rab genes in their genome than the 64 HsRab genes in the human genome. We succeeded in isolating 86 cDNA clones of 88 TtRab genes in the Tetrahymena genome. By comparing the amino acid sequence of Rab in humans and the budding yeast Saccharomyces cerevisiae, 42 TtRab belonged to subfamilies functionally characterized and designated as conventional Rab, while the remaining 44 TtRab were considered to be species-specific. To examine the diversity of Rab in ciliates, we searched for Rab genes in the genome database of P. tetraurelia. Overall, 229 PtRab genes were found and categorized as 157 conventional and 72 species-specific PtRab, respectively. Among them, nine PtRab genes showed high homology to seven TtRab, suggesting the conservation of ciliate-specific Rab. These data suggested that the range of Rab is markedly amplified and diversified in ciliates, which may support the elaborate cellular structures and vigorous phagocytosis of those organisms.

  1. Interface Characteristics and Mechanical Properties of the Vacuum-Brazed Joint of Titanium-Steel Having a Silver-Based Brazing Alloy

    NASA Astrophysics Data System (ADS)

    Elrefaey, A.; Tillmann, W.

    2007-12-01

    The lap joint of a commercially pure titanium plate (CP Ti) to a low-carbon steel plate was produced with a vacuum-brazed furnace using a silver-based filler alloy at different temperatures and lap widths in order to investigate the effects of such brazing parameters on the joint strength and structure. It was found that the shear strength of brazed joints depends strongly on the lap width. Furthermore, we have discovered that the shear strength of the joints increases as the lap width decreases. The maximum shear strength of the joints, 54 MPa, was obtained when they were brazed at 850 °C for 15 minutes. Our observations indicate that intermetallic compound layers of CuTi and Cu2Ti were formed at the interface of the brazed area. An increase in the brazing temperature led to increased growth of these layers. Shear tests show that the fracture path occurred mainly at the CuTi layer, because this layer is hard and brittle.

  2. Evolution of Microstructure in Brazed Joints of Austenitic-Martensitic Stainless Steel with Pure Silver Obtained with Ag-27Cu-5Sn Brazing Filler Material

    NASA Astrophysics Data System (ADS)

    Gangadharan, S.; Sivakumar, D.; Venkateswaran, T.; Kulkarni, Kaustubh

    2016-12-01

    Brazing of an austenitic-martensitic stainless steel (AMSS) with pure silver was carried out at 1053 K, 1073 K, and 1093 K (780 °C, 800 °C, and 820 °C) with Ag-27Cu-5Sn (wt pct) as brazing filler material (BFM). Wettability of the liquid BFM over base AMSS surface was found to be poor. Application of nickel coating to the steel was observed to enhance the wettability and to enable the formation of a good bond between BFM and the steel. The mechanism responsible for enhanced metallurgical bonding of the BFM with AMSS in the presence of nickel coating was explained based on diffusional interactions and uphill diffusion of iron, chromium and nickel observed in the brazed microstructure. Good diffusion-assisted zone was observed to form on silver side at all three temperatures. Four phases were encountered within the joint including silver solid solution, copper solid solution, Cu3Sn intermetallic and Ni-Fe solid solution. The Cu3Sn intermetallic was present in small amounts in the joints brazed at 1053 K and 1073 K (780 °C and 800 °C). The joint formed at 1093 K (820 °C) exhibited the absence of Cu3Sn, fewer defects and larger diffusion-assisted zone. Hardness of base AMSS was found to reduce during brazing due to austenite reversion and post-brazing sub-zero treatment for 2.5 hours was found suitable to recover the hardness.

  3. Dynamic Changes in the Intracellular Association of Selected Rab Small GTPases with MHC Class II and DM during Dendritic Cell Maturation

    PubMed Central

    Pérez-Montesinos, Gibrán; López-Ortega, Orestes; Piedra-Reyes, Jessica; Bonifaz, Laura C.; Moreno, José

    2017-01-01

    Antigen processing for presentation by major histocompatibility complex class II (MHCII) molecules requires the latter to travel through the endocytic pathway together with its chaperons: the invariant chain (Ii) and DM. Nevertheless, the nature of the compartments where MHCII molecules travel to acquire peptides lacks definition regarding molecules involved in intracellular vesicular trafficking, such as Rab small GTPases. We aimed to define which Rab proteins are present during the intracellular transport of MHCII, DM, and Ii through the endocytic pathway on their route to the cell surface during dendritic cell (DC) maturation. We examined, by means of three-color confocal microscopy, the association of MHCII, DM, and Ii with Rab5, Rab7, Rab9, and Rab11 during the maturation of bone marrow-derived or spleen DC in response to LPS as an inflammatory stimulus. Prior to the stage of immature DC, MHCII migrated from diffuse small cytoplasmic vesicles, predominantly Rab5+Rab7− and Rab5+Rab7+ into a pericentriolar Rab5+Rab7+Rab9+ cluster, with Rab11+ areas. As DC reached the mature phenotype, MHCII left the pericentriolar endocytic compartments toward the cell surface in Rab11+ and Rab9+Rab11+ vesicles. The invariant chain and MHCII transport pathways were not identical. DM and MHCII appeared to arrive to pericentriolar endocytic compartments of immature DC through partially different routes. The association of MHCII molecules with distinct Rab GTPases during DC maturation suggests that after leaving the biosynthetic pathway, MHCII sequentially traffic from typical early endosomes to multivesicular late endosomes to finally arrive at the cell surface in Rab11+ recycling-type endosomes. In immature DCs, DM encounters transiently MHCII in the Rab5+Rab7+Rab9+ compartments, to remain there in mature DC.

  4. Al2O3/SUS304 Brazing via AgCuTi-W Composite as Active Filler

    NASA Astrophysics Data System (ADS)

    Su, Cherng-Yuh; Zhuang, Xie-Zongyang; Pan, Cheng-Tang

    2014-03-01

    Alumina ceramic (α-Al2O3) was brazed to stainless steel (SUS304) using an Ag-Cu-Ti + W composite filler and a traditional active brazing filler alloy (CuSil-ABA). Then, the effects of the presence of W particles and of the brazing parameters on the microstructures and mechanical properties of the brazed joints were investigated. The maximum tensile strength of the joints obtained using Ag-Cu-Ti + W composite filler was 13.2 MPa, which is similar to that obtained using CuSil-ABA filler (13.5 MPa). When the joint was brazed at 930 °C for 30 min, the tensile strengths decreased for both kinds of fillers, although the strength was slightly higher for the Ag-Cu-Ti + W composite filler than for the Ag-Cu-Ti filler. The interfacial microstructure results show that the Ti reacts with W to form a Ti-W-O compound in the brazing alloy. When there are more W particles in the brazing alloy, the thickness of the Ti X O Y reaction layer near the alumina ceramic decreases. Moreover, W particles added to the brazing alloy can reduce the coefficient of thermal expansion of the brazing alloy, which results in lower residual stress between the Al2O3 and SUS304 in the brazing joints and thus yields higher tensile strengths as compared to those obtained using the CuSil-ABA brazing alloy.

  5. Heterologous expression of rab4 reduces glucose transport and GLUT4 abundance at the cell surface in oocytes.

    PubMed Central

    Mora, S; Monden, I; Zorzano, A; Keller, K

    1997-01-01

    To evaluate the role of the small rab GTP-binding proteins in glucose transporter trafficking, we have heterologously co-expressed rab4 or rab5 and GLUT4 or GLUT1 glucose transporters in Xenopus oocytes. Co-injection of rab4 and GLUT4 cRNAs resulted in a dose-dependent decrease in glucose transport; this effect was specific for rab4, since co-injection of an inactive rab4 mutant or rab5 cRNA did not have any effect on glucose transport. The effect of rab4 was selective for GLUT4, since no effect was detected in GLUT1-expressing oocytes. The inhibitory effect of rab4 on GLUT4-induced glucose transport was not the result of a change in overall cellular levels of GLUT4 glucose transporters. However, rab4 expression caused a marked decrease in the abundance of GLUT4 transporters present at the cell surface. Finally, rab4 and inhibitors of PtdIns 3-kinase showed additive effects in decreasing glucose transport in GLUT4-expressing oocytes. We conclude that rab4 plays an important role in the regulation of the intracellular GLUT4 trafficking pathway, by contributing to the intracellular retention of GLUT4 through a PtdIns 3-kinase-independent mechanism. PMID:9182703

  6. Development of optimum process parameters and a study of the effects of surface roughness on brazing of copper

    NASA Astrophysics Data System (ADS)

    Zaharinie, Tuan; Huda, Zainul; Izuan, Mohd Faaliq; Hamdi, Mohammed

    2015-03-01

    Brazing experiments on commercially-pure copper plates, using brazing filler metal (MBF-2005), are conducted at temperatures in the range of 650-750 °C for time-durations in the range of 5-15 min. Shear tests for braze-joints involved use of a universal testing machine. Based on the shear-test results, a new brazing cycle has been developed that corresponds to the greatest shear strength of the braze-joint. The brazing cycle has been performed under a controlled dry-argon atmosphere in a tube furnace. Microscopic observations were made by use of both optical and electron microscopes; whereas surface roughness measurements were made by using a TR100 Surface Roughness Tester. It is found that successful brazing and good wetting can be achieved by the least voids by using an average surface roughness (Ra value) for the base material.

  7. Rab10 delivers GLUT4 storage vesicles to the plasma membrane.

    PubMed

    Chen, Yu; Lippincott-Schwartz, Jennifer

    2013-05-01

    The glucose transporter, GLUT4, redistributes to the plasma membrane (PM) upon insulin stimulation, but also recycles through endosomal compartments. Different Rab proteins control these transport itineraries of GLUT4. However, the specific roles played by different Rab proteins in GLUT4 trafficking has been difficult to assess, primarily due to the complexity of endomembrane organization and trafficking. To address this problem, we recently performed advanced live cell imaging using total internal reflection fluorescence (TIRF) microscopy, which images objects ~150 nm from the PM, directly visualizing GLUT4 trafficking in response to insulin stimulation. Using IRAP-pHluorin to selectively label GSVs undergoing PM fusion in response to insulin, we identified Rab10 as the only Rab protein that binds this compartment. Rab14 was found to label transferrin-positive, endosomal compartments containing GLUT4. These also could fuse with the PM in response to insulin, albeit more slowly. Several other Rab proteins, including Rab4A, 4B and 8A, were found to mediate GLUT4 intra-endosomal recycling, serving to internalize surface-bound GLUT4 into endosomal compartments for ultimate delivery to GSVs. Thus, multiple Rab proteins regulate the circulation of GLUT4 molecules within the endomembrane system, maintaining optimal insulin responsiveness within cells.

  8. Role of the small GTPase Rab27a during Herpes simplex virus infection of oligodendrocytic cells

    PubMed Central

    2012-01-01

    Background The morphogenesis of herpes simplex virus type 1 (HSV-1) comprises several events, of which some are not completely understood. It has been shown that HSV-1 glycoproteins accumulate in the trans-Golgi network (TGN) and in TGN-derived vesicles. It is also accepted that HSV-1 acquires its final morphology through a secondary envelopment by budding into TGN-derived vesicles coated with viral glycoproteins and tegument proteins. Nevertheless, several aspects of this process remain elusive. The small GTPase Rab27a has been implicated in regulated exocytosis, and it seems to play a key role in certain membrane trafficking events. Rab27a also seems to be required for human cytomegalovirus assembly. However, despite the involvement of various Rab GTPases in HSV-1 envelopment, there is, to date, no data reported on the role of Rab27a in HSV-1 infection. Results Herein, we show that Rab27a colocalized with GHSV-UL46, a tegument-tagged green fluorescent protein-HSV-1, in the TGN. In fact, this small GTPase colocalized with viral glycoproteins gH and gD in that compartment. Functional analysis through Rab27a depletion showed a significant decrease in the number of infected cells and viral production in Rab27a-silenced cells. Conclusions Altogether, our results indicate that Rab27a plays an important role in HSV-1 infection of oligodendrocytic cells. PMID:23164453

  9. Molecular imaging analysis of Rab GTPases in the regulation of phagocytosis and macropinocytosis.

    PubMed

    Egami, Youhei

    2016-01-01

    Phagocytosis and macropinocytosis, actin-dependent endocytic pathways that mediate the uptake of particles and fluid, respectively, are fundamental routes that enable cells to sample their environment, eliminate pathogens and endogenous cell debris, and contribute to immunoprotection and the maintenance of tissue homeostasis. These processes require a well-organized network of actin cytoskeletal remodeling and membrane transport, which are spatiotemporally regulated by small GTPases. The Rab family of small GTPases, which functions as molecular switches, plays central roles in intracellular membrane trafficking. Although multiple Rab proteins are localized to phagosomes and regulate phagosome maturation, the precise role of each Rab family member in Fcγ receptor (FcγR)-mediated phagocytosis is not fully characterized. Recently, we revealed that Rab35 and Rab20 are important regulators of phagosome formation and maturation, respectively. This review summarizes the functional implication of these Rab GTPases during FcγR-mediated phagocytosis in macrophages. Currently, compared with our knowledge of the regulatory mechanisms of receptor-mediated endocytosis including phagocytosis, the molecular components and signaling cascades of macropinocytosis remain poorly elucidated. Our time-lapse imaging showed that several Rab GTPases are sequentially recruited to the membrane of macropinosomes. Based on our observations, these findings regarding the spatiotemporal localization of Rab GTPases during macropinocytosis are introduced.

  10. Rab and Arl GTPase family members cooperate in the localization of the golgin GCC185.

    PubMed

    Burguete, Alondra Schweizer; Fenn, Timothy D; Brunger, Axel T; Pfeffer, Suzanne R

    2008-01-25

    GCC185 is a large coiled-coil protein at the trans Golgi network that is required for receipt of transport vesicles inbound from late endosomes and for anchoring noncentrosomal microtubules that emanate from the Golgi. Here, we demonstrate that recruitment of GCC185 to the Golgi is mediated by two Golgi-localized small GTPases of the Rab and Arl families. GCC185 binds Rab6, and mutation of residues needed for Rab binding abolishes Golgi localization. The crystal structure of Rab6 bound to the GCC185 Rab-binding domain reveals that Rab6 recognizes a two-fold symmetric surface on a coiled coil immediately adjacent to a C-terminal GRIP domain. Unexpectedly, Rab6 binding promotes association of Arl1 with the GRIP domain. We present a structure-derived model for dual GTPase membrane attachment that highlights the potential ability of Rab GTPases to reach binding partners at a significant distance from the membrane via their unstructured and membrane-anchored, hypervariable domains.

  11. Rab3 is involved in cellular immune responses of the cotton bollworm, Helicoverpa armigera.

    PubMed

    Li, Jie; Song, Cai-Xia; Li, Yu-Ping; Li, Li; Wei, Xiu-Hong; Wang, Jia-Lin; Liu, Xu-Sheng

    2015-06-01

    Rab3, a member of the Rab GTPase family, has been found to be involved in innate immunity. However, the precise function of this GTPase in innate immunity remains unknown. In this study, we identified a Rab3 gene (Ha-Rab3) from the cotton bollworm, Helicoverpa armigera and studied its roles in innate immune responses. Expression of Ha-Rab3 was upregulated in the hemocytes of H. armigera larvae after the injection of Escherichia coli or chromatography beads. The dsRNA-mediated knockdown of Ha-Rab3 gene in H. armigera larval hemocytes led to significant reduction in the phagocytosis and nodulation activities of hemocytes against E. coli, significant increase in the bacterial load in larval hemolymph, and significant reduction in the encapsulation activities of hemocytes toward invading chromatography beads. Furthermore, Ha-Rab3 knockdown significantly suppressed spreading of plasmatocytes. These results suggest that Ha-Rab3 plays important roles in H. armigera cellular immune responses, possibly by mediating spreading of hemocytes.

  12. Rab6 is increased in Alzheimer's disease brain and correlates with endoplasmic reticulum stress.

    PubMed

    Scheper, W; Hoozemans, J J M; Hoogenraad, C C; Rozemuller, A J M; Eikelenboom, P; Baas, F

    2007-10-01

    Alzheimer's disease (AD) is characterized by deposits of aggregated proteins. Accumulation of aggregation-prone proteins activates protein quality control mechanisms, such as the unfolded protein response (UPR) in the endoplasmic reticulum (ER). We previously reported upregulation of the UPR marker BiP in AD brain. In this study, we investigated the small GTPase Rab6, which is involved in retrograde Golgi-ER trafficking and may function as a post-ER quality control system. Using immunohistochemistry and semiquantitative Western blotting, the expression of Rab6 was analysed in hippocampus, entorhinal and temporal cortex of 10 AD patients and six nondemented control subjects. Rab6 is upregulated in AD temporal cortex from Braak stage 3/4, the same stage that UPR activation is found. We observe increased neuronal Rab6 immunoreactivity in all brain areas examined. Although some neurones show colocalization of immunoreactivity for Rab6 and hyperphosphorylated tau, strong Rab6 staining does not colocalize with tangles. We find a highly significant correlation between the Rab6 and BiP levels. In vitro data show that Rab6 is not upregulated as a result of UPR activation or proteasome inhibition indicating an independent regulatory mechanism. Our data suggest that ER and post-ER protein quality control mechanisms are activated early in the pathology of AD.

  13. Rab5a-mediated autophagy regulates the phenotype and behavior of vascular smooth muscle cells

    PubMed Central

    Tan, Jin-Yun; Jia, Luo-Qi; Shi, Wei-Hao; He, Qing; Zhu, Lei; Yu, Bo

    2016-01-01

    Rab5a, a key member of the Rab family of GTPases, was determined to be a regulator of vascular smooth muscle cell (VSMC) proliferation and migration. However, the exact regulatory mechanism remains unclear. As Rab5a has been shown to be associated with autophagy, which is essential for the conversion of VSMCs from a contractile to a synthetic phenotype in order to prevent cell death due to oxidative stress. The present study hypothesized that autophagy may be responsible for the proliferation and migration of VSMCs via the Rab5a protein. The aim of the present study was to evaluate the effect of Rab5a on autophagy in VSMCs. The human aorta vascular smooth muscle cell line, T/G HA-VSMCs, was treated with small interfering (si)RNA against Rab5a and/or platelet-derived growth factor (PDGF). Following treatment, the phenotype transition of the VSMCs was evaluated by detecting the mRNA and protien expression levels of VSMC molecular markers using reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. In addition, autophagy in VSMCs was evaluated by western blotting for autophagy-associated proteins, flow cytometry of acidic vesicular organelles, punctate fluorescence of microtubule associated protein light chain 3 and transmission electron microscopy of typical scattered double-membrane vacuolar structures. Additionally, the proliferation, migration, cell cycle and apoptotic response of VSMCs were detected by sulforhodamine B assay, transwell assay and flow cytometry, respectively. The results revealed that transfection with siRNA against Rab5a led to a significant decrease in Rab5a protein expression, while the reduced expression trend of Rab5a was rescued by intervention with PDGF. Furthermore, cells transfected with siRNA against Rab5a inhibited the autophagy of VSMCs. Downregulated Rab5a inhibited the phenotype transition of VSMCs. Additionally, downregulated Rab5a led to slowed cell growth, decreased numbers of migrated

  14. Graphite to Inconel brazing using active filler metal

    SciTech Connect

    King, J.F.; Baity, F.W.; Walls, J.C.; Hoffman, D.J.

    1989-01-01

    Ion cyclotron resonant frequency (ICRF) antennas are designed to supply large amounts of auxiliary heating power to fusion-grade plasmas in the Toroidal Fusion Test Reactor (TFTR) and Tore Supra fusion energy experiments. A single Faraday shield structure protects a pair of resonant double loops which are designed to launch up to 2 MW of power per loop. The shield consists of two tiers of actively cooled Inconel alloy tubes with the front tier being covered with semicircular graphite tiles. Successful operation of the antenna requires the making of high integrity bonds between the Inconel tubes and graphite tiles by brazing. This paper discusses this process.

  15. Design data for brazed Rene 41 honeycomb sandwich

    NASA Technical Reports Server (NTRS)

    Hepler, A. K.; Arnquist, J.; Koetje, E. L.; Esposito, J. J.; Lindsay, V. E. J.; Swegle, A. R.

    1981-01-01

    Strength data, creep data and residual strength data after cyclic thermal exposure were obtained at temperatures from 78 K to 1144 K (-320 F to 1600 F). The influences of face thickness, core depth, core gage, cell size and thermal/stress exposure conditions on the mechanical design properties were investigated. A braze alloy and process was developed that is adequate to fully develop the strength of the honeycomb core while simultaneously solution treating and aging the Rene 41 fact sheets. New test procedures and test specimen configurations were developed to avoid excessive thermal stresses during cyclic thermal exposure.

  16. Service evaluation of aluminum-brazed titanium (ABTi). [aircraft structures

    NASA Technical Reports Server (NTRS)

    Elrod, S. D.

    1981-01-01

    Long term creep-rupture, flight service and jet engine exhaust tests on aluminum-brazed titanium (ABTi), originally initiated under the DOT/SST follow-on program, were completed. These tests included exposure to natural airline service environments for up to 6 years. The results showed that ABTi has adequate corrosion resistance for long time commercial airplane structural applications. Special precautions are required for those sandwich structures designed for sound attenuation that utilize perforated skins. ABTi was also shown to have usable creep-rupture strength and to be metallurgically stable at temperatures up to 425 C (800 F).

  17. Standard operating procedure: Gas atmosphere MELCO brazing furnace

    SciTech Connect

    Waller, C.R.

    1988-08-01

    A hydrogen and argon gas atmosphere furnace facility using electric furnaces is located at the Clinton P. Anderson Meson Physics Facility (LAMPF). This furnace system was acquired to handle smaller jobs with a more rapid response time than was possible with the larger furnaces. Accelerator- and experimental-related components best assembled by atmosphere brazing techniques are routinely processed by this facility in addition to special heat treatment and bakeout heats. The detailed operation sequence and description of the MELCO furnace system are covered by this report. This document is to augment LA-10231-SOP, which describes the operation of the large furnace systems. 6 figs.

  18. RAB1A promotes Vaccinia virus replication by facilitating the production of intracellular enveloped virions

    PubMed Central

    Pechenick Jowers, Tali; Featherstone, Rebecca J.; Reynolds, Danielle K.; Brown, Helen K.; James, John; Prescott, Alan; Haga, Ismar R.; Beard, Philippa M.

    2015-01-01

    Vaccinia virus (VACV) is a large double-stranded DNA virus with a complex cytoplasmic replication cycle that exploits numerous cellular proteins. This work characterises the role of a proviral cellular protein, the small GTPase RAB1A, in VACV replication. Using siRNA, we identified RAB1A as required for the production of extracellular enveloped virions (EEVs), but not intracellular mature virions (IMVs). Immunofluorescence and electron microscopy further refined the role of RAB1A as facilitating the wrapping of IMVs to become intracellular enveloped virions (IEVs). This is consistent with the known function of RAB1A in maintenance of ER to Golgi transport. VACV can therefore be added to the growing list of viruses which require RAB1A for optimal replication, highlighting this protein as a broadly proviral host factor. PMID:25462347

  19. Lgl1 activation of rab10 promotes axonal membrane trafficking underlying neuronal polarization.

    PubMed

    Wang, Tong; Liu, Yang; Xu, Xiao-Hui; Deng, Cai-Yun; Wu, Kong-Yan; Zhu, Ji; Fu, Xiu-Qing; He, Miao; Luo, Zhen-Ge

    2011-09-13

    Directed membrane trafficking is believed to be crucial for axon development during neuronal morphogenesis. However, the underlying mechanisms are poorly understood. Here, we report a role of Lgl1, the mammalian homolog of Drosophila tumor suppressor Lethal giant larvae, in controlling membrane trafficking underlying axonal growth. We find that Lgl1 is associated with plasmalemmal precursor vesicles and enriched in developing axons. Lgl1 upregulation promoted axonal growth, whereas downregulation attenuated it as well as directional membrane insertion. Interestingly, Lgl1 interacted with and activated Rab10, a small GTPase that mediates membrane protein trafficking, by releasing GDP dissociation inhibitor (GDI) from Rab10. Furthermore, Rab10 lies downstream of Lgl1 in axon development and directional membrane insertion. Finally, both Lgl1 and Rab10 are required for neocortical neuronal polarization in vivo. Thus, the Lgl1 regulation of Rab10 stimulates the trafficking of membrane precursor vesicles, whose fusion with the plasmalemma is crucial for axonal growth.

  20. Phosphorylation Provides a Negative Mode of Regulation for the Yeast Rab GTPase Sec4p

    PubMed Central

    Heger, Christopher D.; Wrann, Christiane D.; Collins, Ruth N.

    2011-01-01

    The Rab family of Ras-related GTPases are part of a complex signaling circuitry in eukaryotic cells, yet we understand little about the mechanisms that underlie Rab protein participation in such signal transduction networks, or how these networks are integrated at the physiological level. Reversible protein phosphorylation is widely used by cells as a signaling mechanism. Several phospho-Rabs have been identified, however the functional consequences of the modification appear to be diverse and need to be evaluated on an individual basis. In this study we demonstrate a role for phosphorylation as a negative regulatory event for the action of the yeast Rab GTPase Sec4p in regulating polarized growth. Our data suggest that the phosphorylation of the Rab Sec4p prevents interactions with its effector, the exocyst component Sec15p, and that the inhibition may be relieved by a PP2A phosphatase complex containing the regulatory subunit Cdc55p. PMID:21931684

  1. Mutant Huntingtin Impairs BDNF Release from Astrocytes by Disrupting Conversion of Rab3a-GTP into Rab3a-GDP

    PubMed Central

    Hong, Yan; Zhao, Ting

    2016-01-01

    Brain-derived neurotrophic factor (BDNF) is essential for neuronal differentiation and survival. We know that BDNF levels decline in the brains of patients with Huntington's disease (HD), a neurodegenerative disease caused by the expression of mutant huntingtin protein (mHtt), and furthermore that administration of BDNF in HD mice is protective against HD neuropathology. BDNF is produced in neurons, but astrocytes are also an important source of BDNF in the brain. Nonetheless, whether mHtt affects astrocytic BDNF in the HD brain remains unknown. Here we investigated astrocytes from HD140Q knock-in mice and uncovered evidence that mHtt decreases BDNF secretion from astrocytes, which is mediated by exocytosis in astrocytes. Our results demonstrate that mHtt associates with Rab3a, a small GTPase localized on membranes of dense-core vesicles, and prevents GTP-Rab3a from binding to Rab3-GAP1, disrupting the conversion of GTP-Rab3a into GDP-Rab3a and thus impairing the docking of BDNF vesicles on plasma membranes of astrocytes. Importantly, overexpression of Rab3a rescues impaired BDNF vesicle docking and secretion from HD astrocytes. Moreover, ATP release and the number of ATP-containing dense-core vesicles docking are decreased in HD astrocytes, suggesting that the exocytosis of dense-core vesicles is impaired by mHtt in HD astrocytes. Further, Rab3a overexpression reduces reactive astrocytes in the striatum of HD140Q knock-in mice. Our results indicate that compromised exocytosis of BDNF in HD astrocytes contributes to the decreased BDNF levels in HD brains and underscores the importance of improving glial function in the treatment of HD. SIGNIFICANCE STATEMENT Huntington's disease (HD) is an inherited neurodegenerative disorder that affects one in every 10,000 Americans. To date, there is no effective treatment for HD, in part because the pathogenic mechanism driving the disease is not fully understood. The dysfunction of astrocytes is known to contribute to the

  2. Mechanical design of ceramic beam tube braze joints for NOvA kicker magnets

    SciTech Connect

    Ader, C.R.; Reilly, R.E.; Wilson, J.H.; /Fermilab

    2010-05-01

    The NO?A Experiment will construct a detector optimized for electron neutrino detection in the existing NuMI neutrino beam. The NuMI beam line is capable of operating at 400 kW of primary beam power and the upgrade will allow up to 700 kW. Ceramic beam tubes are utilized in numerous kicker magnets in different accelerator rings at Fermi National Accelerator Laboratory. Kovar flanges are brazed onto each beam tube end, since kovar and high alumina ceramic have similar expansion curves. The tube, kovar flange, end piece, and braze foil alloy brazing material are stacked in the furnace and then brazed. The most challenging aspect of fabricating kicker magnets in recent years have been making hermetic vacuum seals on the braze joints between the ceramic and flange. Numerous process variables can influence the robustness of conventional metal/ceramic brazing processes. The ceramic-filler metal interface is normally the weak layer when failure does not occur within the ceramic. Differences between active brazing filler metal and the moly-manganese process will be discussed along with the applicable results of these techniques used for Fermilab production kicker tubes.

  3. Brazed Diamond Micropowder Bur Fabricated by Supersonic Frequency Induction Heating for Precision Machining

    NASA Astrophysics Data System (ADS)

    Ma, Bojiang; Lou, Jianpeng; Pang, Qian

    2014-04-01

    The common brazed diamond micropowder bur fabricated in a vacuum furnace produces an even brazing alloy surface. The small brazed diamond grits show low outcropping from the brazing alloy surface, and the chip space between them is small. The bur shows a low grinding efficiency and poor heat dissipation. In this study, a brazed diamond micropowder bur was fabricated by supersonic frequency induction heating. The method afforded a fluctuant surface on the brazing alloy. The brazed diamond grits with an outcropping height distributed uniformly on the fluctuant surface. The fluctuant surface showed a certain chip space. These characteristics of the tool increased the grinding efficiency and decreased the temperature of the grinding arc area. The roughness R a of the ceramic tile surface trimmed by the tool cylinder was between 0.09 and 0.12 μm. In the first 90 min, the decrease in the weight of the ceramic tile ground by the tool cylinder was higher than that ground by the tool fabricated in a vacuum furnace. When the ceramic tile was cylindrically ground, the temperature of the grinding arc area measured using a thermocouple remained below 70 °C.

  4. Brazing stainless steel using a new MBF-series of Ni-Cr-B-Si amorphous brazing foils

    SciTech Connect

    Rabinkin, A.; Wenski, E.; Ribaudo, A.

    1998-02-01

    A group of new high chromium containing amorphous brazing filler metals in foil form has been developed for applications in highly corrosive and/or high temperature environments. These new alloys contain 10--16 wt-% chromium, 1.2--1.6 wt-% boron, i.e., minimal amount needed for amorphability, 7.0--7.5 wt-% silicon 0--5 wt-% molybdenum, and nickel as the balance. Their T{sub S} is within 975--1,030 C (1,787--1,886 F) and T{sub L} is within 1,090--1,140 C (1,994--2,084 F) ranges. The alloys exhibit low erosion of base metal and show no significant detrimental effects on base metal strength because boron concentration is kept to a minimum. It was found that optimal combination of strength, ductility and fatigue resistance of joints is obtained when brazing is combined with high temperature annealing in one extended cycle. The joints after such treatment are practically free of the brittle central eutectic line and the base metals adjoining braze area have only limited and localized segregation of chromium borides. The joint itself is uniform and has a strong and ductile microstructure with only one nickel-chromium-based solid solution phase. The fracture of brazements occurs predominantly in the base metal with the joint ultimate strength higher than the yield strength of the virgin 316L base metal. The brazements have a high corrosion resistance in sea water and water solutions of ammonia and phosphoric acid.

  5. Identification of a Rab GTPase-activating protein cascade that controls recycling of the Rab5 GTPase Vps21 from the vacuole

    PubMed Central

    Rana, Meenakshi; Lachmann, Jens; Ungermann, Christian

    2015-01-01

    Transport within the endocytic pathway depends on a consecutive function of the endosomal Rab5 and the late endosomal/lysosomal Rab7 GTPases to promote membrane recycling and fusion in the context of endosomal maturation. We previously identified the hexameric BLOC-1 complex as an effector of the yeast Rab5 Vps21, which also recruits the GTPase-activating protein (GAP) Msb3. This raises the question of when Vps21 is inactivated on endosomes. We provide evidence for a Rab cascade in which activation of the Rab7 homologue Ypt7 triggers inactivation of Vps21. We find that the guanine nucleotide exchange factor (GEF) of Ypt7 (the Mon1-Ccz1 complex) and BLOC-1 both localize to the same endosomes. Overexpression of Mon1-Ccz1, which generates additional Ypt7-GTP, or overexpression of activated Ypt7 promotes relocalization of Vps21 from endosomes to the endoplasmic reticulum (ER), which is indicative of Vps21 inactivation. This ER relocalization is prevented by loss of either BLOC-1 or Msb3, but it also occurs in mutants lacking endosome–vacuole fusion machinery such as the HOPS tethering complex, an effector of Ypt7. Importantly, BLOC-1 interacts with the HOPS on vacuoles, suggesting a direct Ypt7-dependent cross-talk. These data indicate that efficient Vps21 recycling requires both Ypt7 and endosome–vacuole fusion, thus suggesting extended control of a GAP cascade beyond Rab interactions. PMID:25971802

  6. Aberrant amino acid signaling promotes growth and metastasis of hepatocellular carcinomas through Rab1A-dependent activation of mTORC1 by Rab1A

    PubMed Central

    Yang, Yang; Zhang, Mei-Yin; Rao, Hui-Lan; Wang, Hui-Yun; Zheng, X.F. Steven

    2015-01-01

    mTORC1 is a master regulator of cell growth and proliferation, and an established anticancer drug target. Aberrant mTORC1 signaling is common in hepatocellular carcinoma (HCC), but the underlying mechanism remains elusive. Rab1A is a newly identified mTORC1 activator that mediates an alternative amino acid (AA) signaling branch to Rag GTPases. Because liver is a physiological hub for nutrient sensing and metabolic homeostasis, we investigated the possible role of Rab1A in HCC. We found that Rab1A is frequently overexpressed in HCC, which enhances hyperactive AA-mTORC1 signaling, promoting malignant growth and metastasis of HCC in vitro and in vivo. Moreover, aberrant Rab1A expression is closely associated with poor prognosis. Strikingly, aberrant Rab1A overexpression leads to increased rapamycin sensitivity, indicating that inappropriate activation of AA signaling is a cancer-driving event in HCC. Our findings further suggest that Rab1A is a valuable biomarker for prognosis and personalized mTORC1-targeted therapy in liver cancer. PMID:26308575

  7. The Rab GTPase Rab8 as a shared regulator of ciliogenesis and immune synapse assembly: From a conserved pathway to diverse cellular structures

    PubMed Central

    Patrussi, Laura; Baldari, Cosima T.

    2016-01-01

    ABSTRACT Rab GTPases, which form the largest branch of the Ras GTPase superfamily, regulate almost every step of vesicle-mediated trafficking. Among them, Rab8 is an essential participant in primary cilium formation. In a report recently published in the Journal of Cell Science, Finetti and colleagues identify Rab8 as a novel player in vesicular traffic in the non-ciliated T lymphocytes, which contributes to the assembly of the specialized signaling platform known as the immune synapse. By interacting with the v-SNARE VAMP-3, Rab8 is indeed responsible for the final docking/fusion step in T cell receptor (TCR) recycling to the immune synapse. A second important take-home message which comes to light from this work is that VAMP-3 also interacts with Rab8 at the base of the cilium in NIH-3T3 cells, where it regulates ciliary growth and targeting of Smoothened at the plasma membrane. Hence the data presented in this report, in addition to identifying Rab8 as a novel player in vesicular traffic to the immune synapse, reveal how both ciliated and non-ciliated cells take advantage of a conserved pathway to build highly specific cellular structures. PMID:26587735

  8. Corrosion Mechanisms in Brazed Al-Base Alloy Sandwich Structures as a Function of Braze Alloy and Process Variables

    DTIC Science & Technology

    2013-02-01

    aeration solution for 8 hours. A concentrated Nitric acid (HNO3) dip for 15 seconds removed corrosion products prior to post-exposure SEM imaging [25...32 to -37°C under a liquid nitrogen chill at 11.2 V for one minute [10]. The electropolishing solution was a mixture of 1/3 concentrated Nitric acid ...DATES COVERED (From - To) 03/27/06-12/31/12 4. TITLE AND SUBTITLE Corrosion Mechanisms in Brazed Al-Base Alloy Sandwich Structures as a Function

  9. Diffusion bonding and brazing of high purity copper for linear collider accelerator structures

    NASA Astrophysics Data System (ADS)

    Elmer, J. W.; Klingmann, J.; van Bibber, K.

    2001-05-01

    Diffusion bonding and brazing of high purity copper were investigated to develop procedures for joining precision machined copper components for the Next Linear Collider (NLC). Diffusion bonds were made over a range of temperatures from 400 °C to 1000 °C, under two different loading conditions [3.45 kPa (0.5 psi) and 3.45 MPa (500 psi)], and on two different diamond machined surface finishes. Brazes were made using pure silver, pure gold, and gold-nickel alloys, and different heating rates produced by both radiation and induction heating. Braze materials were applied by both physical vapor deposition (PVD) and conventional braze alloy shims. Results of the diffusion bonding experiments showed that bond strengths very near that of the copper base metal could be made at bonding temperatures of 700 °C or higher at 3.45 MPa bonding pressure. At lower temperatures, only partial strength diffusion bonds could be made. At low bonding pressures (3.45 kPa), full strength bonds were made at temperatures of 800 °C and higher, while no bonding (zero strength) was observed at temperatures of 700 °C and lower. Observations of the fracture surfaces of the diffusion bonded samples showed the effects of surface finish on the bonding mechanism. These observations clearly indicate that bonding began by point asperity contact, and flatter surfaces resulted in a higher percentage of bonded area under similar bonding conditions. Results of the brazing experiments indicated that pure silver worked very well for brazing under both conventional and high heating rate scenarios. Similarly, pure silver brazed well for both the PVD layers and the braze alloy shims. The gold and gold-containing brazes had problems, mainly due to the high diffusivity of gold in copper. These problems led to the necessity of overdriving the temperature to ensure melting, the presence of porosity in the joint, and very wide braze joints. Based on the overall findings of this study, a two-step joining method

  10. Brazing of refractory, superalloy, and composite materials for Space Shuttle applications.

    NASA Technical Reports Server (NTRS)

    Beuyukian, C. S.

    1972-01-01

    Research work concerning the metallic portion of the shuttle-orbiter heat shield (expected to experience temperatures up to 2500 F) is described. The five metals being evaluated are TD-Ni-Cr, Cb-C129Y, Cb752, Hayes 188, and Inconel 625. Brazing techniques whereby pairs of these materials are joined into thin-membered assemblies for heat shield applications are described. Results obtained with the vacuum-furnace brazing process are examined. In addition, the use of brazed aluminum-boron metal-matrix-contoured composite structures for heat shield applications is evaluated.

  11. Joining C/C composite to copper using active Cu-3.5Si braze

    NASA Astrophysics Data System (ADS)

    Shen, Yuanxun; Li, Zhenglin; Hao, Chuanyong; Zhang, Jinsong

    2012-02-01

    A simple technique was developed to join C/C composite to Cu using active Cu-3.5Si braze for nuclear thermal applications. The brazing alloy exhibited good wettability on C/C substrate due to the reaction layer formed at the interface. A strong interfacial bond of the brazing alloy on C/C with the formation of TiC + SiC + Ti 5Si 3 reaction layer was obtained. The produced CC/Cu/CuCrZr joint exhibited shear strength as high as 79 MPa and excellent thermal resistance during the thermal shock tests.

  12. The causes of high power diode laser brazed seams fractures of dissimilar materials

    NASA Astrophysics Data System (ADS)

    Adamiak, Marcin; Czupryński, Artur; Janicki, Damian; Górka, Jacek

    2016-12-01

    Presented in this article are the results of experiments carried out to determine the causes of braze cracking of dissimilar materials brazed with a ROFIN DL 020 high power diode laser with the use of additional powdered EN AW-1070A aluminium alloy to bond thin aluminium sheets with soft, low alloy DC04+ZE75/75 steel plate which was electrolytically coated with zinc on both sides. Presented are the results of metallographic, macroscopic, microscopic, diffractometric phase analyses of the weld joints. Metallurgical problems arising during processing as well as suggestions regarding technical aspects of laser brazing dissimilar materials in regards to their physical characteristics and chemical composition are explored.

  13. Flaw Tolerance In Lap Shear Brazed Joints. Part 2

    NASA Technical Reports Server (NTRS)

    Wang, Len; Flom, Yury

    2003-01-01

    This paper presents results of the second part of an on-going effort to gain better understanding of defect tolerance in braze joints. In the first part of this three-part series, we mechanically tested and modeled the strength of the lap joints as a function of the overlap distance. A failure criterion was established based on the zone damage theory, which predicts the dependence of the lap joint shear strength on the overlap distance, based on the critical size of a finite damage zone or an overloaded region in the joint. In this second part of the study, we experimentally verified the applicability of the damage zone criterion on prediction of the shear strength of the lap joint and introduced controlled flaws into the lap joints. The purpose of the study was to evaluate the lap joint strength as a function of flaw size and its location through mechanical testing and nonlinear finite element analysis (FEA) employing damage zone criterion for definition of failure. The results obtained from the second part of the investigation confirmed that the failure of the ductile lap shear brazed joints occurs when the damage zone reaches approximately 10% of the overlap width. The same failure criterion was applicable to the lap joints containing flaws.

  14. Evaluation of brazed silicon nitride joints: Microstructure and mechanical properties

    SciTech Connect

    Peteves, S.D.; Nicholas, M.G.

    1996-06-01

    Sintered Si{sub 3}N{sub 4} has been bonded to itself and to AISI 316 steel by the active-metal brazing route. A commercial Ag-35Cu-1.6Ti filler has been used with joining taking place during a 30 min hold at 850 C under vacuum. Si{sub 3}N{sub 4}/Si{sub 3}N{sub 4} joints have been produced with strength distribution (average bend strength = 773.5 MPa, Weibull modulus = 11.2) similar to that of the monolithic ceramic. Direct brazing of the Si{sub 3}N{sub 4} to AISI 316 steel was unsuccessful. However, reliably strong (bend strength of 250--400 MPa) ceramic/steel joints with 20 x 20 mm{sup 2} cross sections were fabricated by using Cu, Mo, or Nb interlayers. The most potent interlayer used in this work was Mo, whose coefficient of thermal expansion matches best that of the ceramic.

  15. A Compendium of Brazed Microstructures For Fission Power Systems Applications

    NASA Technical Reports Server (NTRS)

    Locci, Ivan E.; Bowman, Cheryl L.

    2012-01-01

    NASA has been supporting design studies and technology development for fission-based power systems that could provide power to an outpost on the Moon, Mars, or an asteroid. Technology development efforts have included fabrication and evaluation of components used in a Stirling engine power conversion system. This investigation is part of the development of several braze joints crucial for the heat exchanger transfer path from a hot-side heat exchanger to a Stirling engine heat acceptor. Dissimilar metal joints are required to impart both mechanical strength and thermal path integrity for a heater head of interest. Preliminary design work for the heat exchanger involved joints between low carbon stainless steel to Inconel 718, where the 316L stainless steel would contain flowing liquid metal NaK while Inconel 718, a stronger alloy, would be used as structural reinforcement. This paper addressed the long-term microstructural stability of various braze alloys used to join 316L stainless steel heater head to the high conductivity oxygen-free copper acceptor to ensure the endurance of the critical metallic components of this sophisticated heat exchanger. The bonding of the 316L stainless steel heater head material to a copper heat acceptor is required to increase the heat-transfer surface area in contact with flowing He, which is the Stirling engine working fluid.

  16. Spatial-Temporal Study of Rab1b Dynamics and Function at the ER-Golgi Interface.

    PubMed

    Martinez, Hernán; García, Iris A; Sampieri, Luciana; Alvarez, Cecilia

    2016-01-01

    The GTPase Rab1b is involved in ER to Golgi transport, with multiple Rab1b effectors (located at ERES, VTCs and the Golgi complex) being required for its function. In this study, we performed live-cell dual-expression studies to analyze the dynamics of Rab1b and some effectors located at the ERES-Golgi interface. Rab1b occupied widely distributed mobile punctate and tubular structures, displaying a transient overlaps with its effectors and showing that these overlaps occurred at the same time in spatially distinct steps of ER to Golgi transport. In addition, we assessed Rab1b dynamics during cargo sorting by analyzing the concentration at ERES of a Golgi protein (SialT2-CFP) during Brefeldin A washout (BFA WO). Rab1b was associated to most of the ERES structures, but at different times during BFA WO, and recurrently SialT2-CFP was sorted in the ERES-Rab1b positive structures. Furthermore, we reveal for first time that Rab1b localization time at ERES depended on GBF1, a Rab1b effector that acts as the guanine nucleotide exchange factor of Arf1, and that Rab1b membrane association/dissociation dynamics at ERES was dependent on the GBF1 membrane association and activity, which strongly suggests that GBF1 activity modulates Rab1b membrane cycling dynamic.

  17. Spatial-Temporal Study of Rab1b Dynamics and Function at the ER-Golgi Interface

    PubMed Central

    Martinez, Hernán; García, Iris A.; Sampieri, Luciana

    2016-01-01

    The GTPase Rab1b is involved in ER to Golgi transport, with multiple Rab1b effectors (located at ERES, VTCs and the Golgi complex) being required for its function. In this study, we performed live-cell dual-expression studies to analyze the dynamics of Rab1b and some effectors located at the ERES-Golgi interface. Rab1b occupied widely distributed mobile punctate and tubular structures, displaying a transient overlaps with its effectors and showing that these overlaps occurred at the same time in spatially distinct steps of ER to Golgi transport. In addition, we assessed Rab1b dynamics during cargo sorting by analyzing the concentration at ERES of a Golgi protein (SialT2-CFP) during Brefeldin A washout (BFA WO). Rab1b was associated to most of the ERES structures, but at different times during BFA WO, and recurrently SialT2-CFP was sorted in the ERES-Rab1b positive structures. Furthermore, we reveal for first time that Rab1b localization time at ERES depended on GBF1, a Rab1b effector that acts as the guanine nucleotide exchange factor of Arf1, and that Rab1b membrane association/dissociation dynamics at ERES was dependent on the GBF1 membrane association and activity, which strongly suggests that GBF1 activity modulates Rab1b membrane cycling dynamic. PMID:27500526

  18. The novel Rab11-FIP/Rip/RCP family of proteins displays extensive homo- and hetero-interacting abilities.

    PubMed

    Wallace, Deborah M; Lindsay, Andrew J; Hendrick, Alan G; McCaffrey, Mary W

    2002-04-12

    The Rab11-FIP/Rip/RCP proteins are a recently described novel protein family, whose members interact with Rab GTPases that function in endosomal recycling. To date, five such proteins have been described in humans, all of which interact with Rab11, and one (RCP) also interacts with Rab4. Here, we characterise several of these proteins with respect to their ability to interact with Rab4, as well as their ability to self-interact, and to interact with each other. We now demonstrate that two of the family members-pp75/Rip11 and Rab11-FIP3 do not bind Rab4 and show that several members of the family can self-interact and interact with each other. These interactions primarily involve their C-terminal end which includes the Rab binding domain (RBD) that is contained within a predicted coiled-coil, or ERM motif. We identify a new (sixth) member of the protein family, which we propose to name Rab11-FIP4, and report the family evolutionary complexity and chromosomal distribution. Furthermore, we propose that the ability of these proteins to bind each other will be important in effecting membrane trafficking events by forming protein 'platforms,' regulated by Rab11 and/or Rab4 activity.

  19. A Role for Na+,K+-ATPase α1 in Regulating Rab27a Localisation on Melanosomes

    PubMed Central

    Booth, Antonia E. G.; Tarafder, Abul K.; Hume, Alistair N.; Recchi, Chiara; Seabra, Miguel C.

    2014-01-01

    The mechanism(s) by which Rab GTPases are specifically recruited to distinct intracellular membranes remains elusive. Here we used Rab27a localisation onto melanosomes as a model to investigate Rab targeting. We identified the α1 subunit of Na+,K+-ATPase (ATP1a1) as a novel Rab27a interacting protein in melanocytes and showed that this interaction is direct with the intracellular M4M5 loop of ATP1a1 and independent of nucleotide bound status of the Rab. Knockdown studies in melanocytes revealed that ATP1a1 plays an essential role in Rab27a-dependent melanosome transport. Specifically, expression of ATP1a1, like the Rab27a GDP/GTP exchange factor (Rab3GEP), is essential for targeting and activation of Rab27a to melanosomes. Finally, we showed that the ability of Rab27a mutants to target to melanosomes correlates with the efficiency of their interaction with ATP1a1. Altogether these studies point to a new role for ATP1a1 as a regulator of Rab27a targeting and activation. PMID:25051489

  20. Expression of Rab1A is upregulated in human lung cancer and associated with tumor size and T stage

    PubMed Central

    Qin, Xiaoyu; Huang, Tinglei; Huang, Bo; Zhang, Yanjie; Jiang, Bin

    2016-01-01

    Rab1A expression is associated with malignant phenotypes in several human tumors; however, the role of Rab1A in lung cancer is still unclear. In this study, we attempted to establish the role of Rab1A in major human lung cancer subtypes. Rab1A expression in different histological types of human lung cancer was analyzed in lung cancer tissues with paired adjacent noncancerous tissues and a large panel of lung cancer cell lines. The effect of Rab1A expression on multiple cancer-associated signaling pathways was also examined. The results demonstrated that Rab1A was significantly overexpressed in the different histological types of lung cancer as compared to non-cancerous tissues, and Rab1A expression was correlated with tumor volume and stage. In a large panel of lung cancer cell lines, high Rab1A expression was observed as compared to a normal lung/bronchus epithelial cell line. However, Rab1A protein levels were not correlated with mTORC1 (P-S6K1), mTORC2 (P-AKT), MEK (P-ERK), JNK (P-c-Jun) or p38MAPK (P-MK2) signaling. Rab1A knockdown had no effect on mTOR signaling or cell growth. These data suggested that Rab1A may be involved in the pathogenesis of human lung cancer in an mTOR- and MAPK-independent manner. PMID:27902464

  1. Starvation-induced MTMR13 and RAB21 activity regulates VAMP8 to promote autophagosome-lysosome fusion.

    PubMed

    Jean, Steve; Cox, Sarah; Nassari, Sonya; Kiger, Amy A

    2015-03-01

    Autophagy, the process for recycling cytoplasm in the lysosome, depends on membrane trafficking. We previously identified Drosophila Sbf as a Rab21 guanine nucleotide exchange factor (GEF) that acts with Rab21 in endosomal trafficking. Here, we show that Sbf/MTMR13 and Rab21 have conserved functions required for starvation-induced autophagy. Depletion of Sbf/MTMR13 or Rab21 blocked endolysosomal trafficking of VAMP8, a SNARE required for autophagosome-lysosome fusion. We show that starvation induces Sbf/MTMR13 GEF and RAB21 activity, as well as their induced binding to VAMP8 (or closest Drosophila homolog, Vamp7). MTMR13 is required for RAB21 activation, VAMP8 interaction and VAMP8 endolysosomal trafficking, defining a novel GEF-Rab-effector pathway. These results identify starvation-responsive endosomal regulators and trafficking that tunes membrane demands with changing autophagy status.

  2. Yearlong evaluation of performance and durability of a pilot-scale Revolving Algal Biofilm (RAB) cultivation system.

    PubMed

    Gross, Martin; Wen, Zhiyou

    2014-11-01

    Current algal cultivation has been mainly performed in open ponds or photobioreactors in which algal cells are suspended and harvested through flocculation and centrifugation. A unique attachment based Revolving Algal Biofilm (RAB) cultivation system was recently developed for easy biomass harvest with enhanced biomass productivity. The objective of this research was to evaluate the performance (durability, algal growth, and the geometry) of the RAB system at pilot-scale. A yearlong test of the RAB system was successfully conducted at a greenhouse facility at Boone, Iowa, USA. The RAB resulted in an average of 302% increase in biomass productivity compared to a standard raceway pond, with a maximum biomass productivity (ash free) of 18.9 g/m(2)-day being achieved. The RAB with a vertical configuration generated higher productivity than the triangular RAB. Collectively, the research shows that the RAB as an efficient algal culture system has great potential for being deployed at commercial scale.

  3. The interplay between the Rab27A effectors Slp4-a and MyRIP controls hormone-evoked Weibel-Palade body exocytosis.

    PubMed

    Bierings, Ruben; Hellen, Nicola; Kiskin, Nikolai; Knipe, Laura; Fonseca, Ana-Violeta; Patel, Bijal; Meli, Athina; Rose, Marlene; Hannah, Matthew J; Carter, Tom

    2012-09-27

    Weibel-Palade body (WPB) exocytosis underlies hormone-evoked VWF secretion from endothelial cells (ECs). We identify new endogenous components of the WPB: Rab3B, Rab3D, and the Rab27A/Rab3 effector Slp4-a (granuphilin), and determine their role in WPB exocytosis. We show that Rab3B, Rab3D, and Rab27A contribute to Slp4-a localization to WPBs. siRNA knockdown of Slp4-a, MyRIP, Rab3B, Rab3D, Rab27A, or Rab3B/Rab27A, or overexpression of EGFP-Slp4-a or EGFP-MyRIP showed that Slp4-a is a positive and MyRIP a negative regulator of WPB exocytosis and that Rab27A alone mediates these effects. We found that ECs maintain a constant amount of cellular Rab27A irrespective of the WPB pool size and that Rab27A (and Rab3s) cycle between WPBs and a cytosolic pool. The dynamic redistribution of Rab proteins markedly decreased the Rab27A concentration on individual WPBs with increasing WPB number per cell. Despite this, the probability of WPB release was independent of WPB pool size showing that WPB exocytosis is not determined simply by the absolute amount of Rab27A and its effectors on WPBs. Instead, we propose that the probability of release is determined by the fractional occupancy of WPB-Rab27A by Slp4-a and MyRIP, with the balance favoring exocytosis.

  4. RNAi KNOCKDOWN OF BmRab3 LED TO LARVA AND PUPA LETHALITY IN SILKWORM Bombyx mori L.

    PubMed

    Singh, Chabungbam Orville; Xin, Hu-hu; Chen, Rui-ting; Wang, Mei-xian; Liang, Shuang; Lu, Yan; Cai, Zi-zheng; Zhang, Deng-pan; Miao, Yun-gen

    2015-06-01

    Rab3 GTPases are known to play key a role in vesicular trafficking, and express highest in brain and endocrine tissues. In mammals, Rab3 GTPases are paralogs unlike in insect. In this study, we cloned Rab3 from the silk gland tissue of silkworm Bombyx mori, and identified it as BmRab3. Our in silico analysis indicated that BmRab3 is an isoform with a theoretical isoelectric point and molecular weight of 5.52 and 24.3 kDa, respectively. Further, BmRab3 showed the C-terminal hypervariability for GGT2 site but having two other putative guanine nucleotide exchange factor/GDP dissociation inhibitor interaction sites. Multiple alignment sequence indicated high similarities of BmRab3 with Rab3 isoforms of other species. The phylogeny tree showed BmRab3 clustered between the species of Tribolium castaneum and Aedes aegypti. Meanwhile, the expression analysis of BmRab3 showed the highest expression in middle silk glands (MSGs) than all other tissues in the third day of fifth-instar larva. Simultaneously, we showed the differential expression of BmRab3 in the early instar larva development, followed by higher expression in male than female pupae. In vivo dsRNA interference of BmRab3 reduced the expression of BmRab3 by 75% compared to the control in the MSGs in the first day. But as the worm grew to the third day, the difference of BmRab3 between knockdown and control was only about 10%. The knockdown later witnessed underdevelopment of the larvae and pharate pupae lethality in the overall development of silkworm B. mori L.

  5. Rab3A, a possible marker of cortical granules, participates in cortical granule exocytosis in mouse eggs.

    PubMed

    Bello, Oscar Daniel; Cappa, Andrea Isabel; de Paola, Matilde; Zanetti, María Natalia; Fukuda, Mitsunori; Fissore, Rafael A; Mayorga, Luis S; Michaut, Marcela A

    2016-09-10

    Fusion of cortical granules with the oocyte plasma membrane is the most significant event to prevent polyspermy. This particular exocytosis, also known as cortical reaction, is regulated by calcium and its molecular mechanism is still not known. Rab3A, a member of the small GTP-binding protein superfamily, has been implicated in calcium-dependent exocytosis and is not yet clear whether Rab3A participates in cortical granules exocytosis. Here, we examine the involvement of Rab3A in the physiology of cortical granules, particularly, in their distribution during oocyte maturation and activation, and their participation in membrane fusion during cortical granule exocytosis. Immunofluorescence and Western blot analysis showed that Rab3A and cortical granules have a similar migration pattern during oocyte maturation, and that Rab3A is no longer detected after cortical granule exocytosis. These results suggested that Rab3A might be a marker of cortical granules. Overexpression of EGFP-Rab3A colocalized with cortical granules with a Pearson correlation coefficient of +0.967, indicating that Rab3A and cortical granules have almost a perfect colocalization in the egg cortical region. Using a functional assay, we demonstrated that microinjection of recombinant, prenylated and active GST-Rab3A triggered cortical granule exocytosis, indicating that Rab3A has an active role in this secretory pathway. To confirm this active role, we inhibited the function of endogenous Rab3A by microinjecting a polyclonal antibody raised against Rab3A prior to parthenogenetic activation. Our results showed that Rab3A antibody microinjection abolished cortical granule exocytosis in parthenogenetically activated oocytes. Altogether, our findings confirm that Rab3A might function as a marker of cortical granules and participates in cortical granule exocytosis in mouse eggs.

  6. Rab25 acts as an oncogene in luminal B breast cancer and is causally associated with Snail driven EMT

    PubMed Central

    Mitra, Shreya; Federico, Lorenzo; Zhao, Wei; Dennison, Jennifer; Sarkar, Tapasree Roy; Zhang, Fan; Takiar, Vinita; Cheng, Kwai W.; Mani, Sendurai; Lee, Ju Seog; Mills, Gordon B.

    2016-01-01

    The Rab GTPases regulate vesicular trafficking machinery that transports and delivers a diverse pool of cargo, including growth factor receptors, integrins, nutrient receptors and junction proteins to specific intracellular sites. The trafficking machinery is indeed a major posttranslational modifier and is critical for cellular homeostasis. Deregulation of this stringently controlled system leads to a wide spectrum of disorders including cancer. Herein we demonstrate that Rab25, a key GTPase, mostly decorating the apical recycling endosome, is a dichotomous variable in breast cancer cell lines with higher mRNA and protein expression in Estrogen Receptor positive (ER+ve) lines. Rab25 and its effector, Rab Coupling Protein (RCP) are frequently coamplified and coordinately elevated in ER+ve breast cancers. In contrast, Rab25 levels are decreased in basal-like and almost completely lost in claudin-low tumors. This dichotomy exists despite the presence of the 1q amplicon that hosts Rab25 across breast cancer subtypes and is likely due to differential methylation of the Rab25 promoter. Functionally, elevated levels of Rab25 drive major hallmarks of cancer including indefinite growth and metastasis but in case of luminal B breast cancer only. Importantly, in such ER+ve tumors, coexpression of Rab25 and its effector, RCP is significantly associated with a markedly worsened clinical outcome. Importantly, in claudin-low cell lines, exogenous Rab25 markedly inhibits cell migration. Similarly, during Snail-induced epithelial to mesenchymal transition (EMT) exogenous Rab25 potently reverses Snail-driven invasion. Overall, this study substantiates a striking context dependent role of Rab25 in breast cancer where Rab25 is amplified and enhances aggressiveness in luminal B cancers while in claudin-low tumors, Rab25 is lost indicating possible anti-tumor functions. PMID:27259233

  7. Rab1 recruits WHAMM during membrane remodeling but limits actin nucleation.

    PubMed

    Russo, Ashley J; Mathiowetz, Alyssa J; Hong, Steven; Welch, Matthew D; Campellone, Kenneth G

    2016-03-15

    Small G-proteins are key regulatory molecules that activate the actin nucleation machinery to drive cytoskeletal rearrangements during plasma membrane remodeling. However, the ability of small G-proteins to interact with nucleation factors on internal membranes to control trafficking processes has not been well characterized. Here we investigated roles for members of the Rho, Arf, and Rab G-protein families in regulating WASP homologue associated with actin, membranes, and microtubules (WHAMM), an activator of Arp2/3 complex-mediated actin nucleation. We found that Rab1 stimulated the formation and elongation of WHAMM-associated membrane tubules in cells. Active Rab1 recruited WHAMM to dynamic tubulovesicular structures in fibroblasts, and an active prenylated version of Rab1 bound directly to an N-terminal domain of WHAMM in vitro. In contrast to other G-protein-nucleation factor interactions, Rab1 binding inhibited WHAMM-mediated actin assembly. This ability of Rab1 to regulate WHAMM and the Arp2/3 complex represents a distinct strategy for membrane remodeling in which a Rab G-protein recruits the actin nucleation machinery but dampens its activity.

  8. Rab27A regulates exosome secretion from lung adenocarcinoma cells A549: involvement of EPI64.

    PubMed

    Li, Wenhai; Hu, Yunsheng; Jiang, Tao; Han, Yong; Han, Guoliang; Chen, Jiakuan; Li, Xiaofei

    2014-11-01

    Exosomes are small membrane vesicles secreted into the extracellular compartment by exocytosis. The unique composition of exosomes can be transported to other cells which allow cells to exert biological functions at distant sites. However, in lung cancer, the regulation of exosome secretion was poorly understood. In this study, we employed human lung adenocarcinoma A549 cells to determine the exosome secretion and involved regulation mechanism. We found that Rab27A was expressed in A549 cells and the reduction of Rab27A by Rab27A-specific shRNA could significantly decrease the secretion of exosome by A549 cells. EPI64, a candidate GAP that is specific for Rab27, was also detected in A549 cells. By pull-down assay, we found that EPI64 participated in the exosome secretion of A549 cells by acting as a specific GAP for Rab27A, not Rab27B. Overexpression of EPI64 enhanced exosome secretion. Taken together, in A549 cells, EPI64 could regulate the exosome secretion by functioning as a GAP specific for Rab27A.

  9. A Bacterial Pathogen Targets a Host Rab-Family GTPase Defense Pathway with a GAP.

    PubMed

    Spanò, Stefania; Gao, Xiang; Hannemann, Sebastian; Lara-Tejero, María; Galán, Jorge E

    2016-02-10

    Cell-autonomous defense mechanisms are potent strategies that protect individual cells against intracellular pathogens. The Rab-family GTPase Rab32 was previously shown to restrict the intracellular human pathogen Salmonella Typhi, but its potential broader role in antimicrobial defense remains unknown. We show that Rab32 represents a general cell-autonomous, antimicrobial defense that is counteracted by two Salmonella effectors. Mice lacking Rab-32 or its nucleotide exchange factor BLOC-3 are permissive to S. Typhi infection and exhibit increased susceptibility to S. Typhimurium. S. Typhimurium counters this defense pathway by delivering two type III secretion effectors, SopD2, a Rab32 GAP, and GtgE, a specific Rab32 protease. An S. Typhimurium mutant strain lacking these two effectors exhibits markedly reduced virulence, which is fully restored in BLOC-3-deficient mice. These results demonstrate that a cell-autonomous, Rab32-dependent host defense pathway plays a central role in the defense against vacuolar pathogens and describe a mechanism evolved by a bacterial pathogen to counter it.

  10. Rab8b Regulates Transport of West Nile Virus Particles from Recycling Endosomes.

    PubMed

    Kobayashi, Shintaro; Suzuki, Tadaki; Kawaguchi, Akira; Phongphaew, Wallaya; Yoshii, Kentaro; Iwano, Tomohiko; Harada, Akihiro; Kariwa, Hiroaki; Orba, Yasuko; Sawa, Hirofumi

    2016-03-18

    West Nile virus (WNV) particles assemble at and bud into the endoplasmic reticulum (ER) and are secreted from infected cells through the secretory pathway. However, the host factor related to these steps is not fully understood. Rab proteins, belonging to the Ras superfamily, play essential roles in regulating many aspects of vesicular trafficking. In this study, we sought to determine which Rab proteins are involved in intracellular trafficking of nascent WNV particles. RNAi analysis revealed that Rab8b plays a role in WNV particle release. We found that Rab8 and WNV antigen were colocalized in WNV-infected human neuroblastoma cells, and that WNV infection enhanced Rab8 expression in the cells. In addition, the amount of WNV particles in the supernatant of Rab8b-deficient cells was significantly decreased compared with that of wild-type cells. We also demonstrated that WNV particles accumulated in the recycling endosomes in WNV-infected cells. In summary, these results suggest that Rab8b is involved in trafficking of WNV particles from recycling endosomes to the plasma membrane.

  11. Rab35 regulates cadherin-mediated adherens junction formation and myoblast fusion

    PubMed Central

    Charrasse, Sophie; Comunale, Franck; De Rossi, Sylvain; Echard, Arnaud; Gauthier-Rouvière, Cécile

    2013-01-01

    Cadherins are homophilic cell–cell adhesion molecules implicated in many fundamental processes, such as morphogenesis, cell growth, and differentiation. They accumulate at cell–cell contact sites and assemble into large macromolecular complexes named adherens junctions (AJs). Cadherin targeting and function are regulated by various cellular processes, many players of which remain to be uncovered. Here we identify the small GTPase Rab35 as a new regulator of cadherin trafficking and stabilization at cell–cell contacts in C2C12 myoblasts and HeLa cells. We find that Rab35 accumulates at cell–cell contacts in a cadherin-dependent manner. Knockdown of Rab35 or expression of a dominant-negative form of Rab35 impaired N- and M-cadherin recruitment to cell–cell contacts, their stabilization at the plasma membrane, and association with p120 catenin and led to their accumulation in transferrin-, clathrin-, and AP-2–positive intracellular vesicles. We also find that Rab35 function is required for PIP5KIγ accumulation at cell–cell contacts and phosphatidyl inositol 4,5-bisphosphate production, which is involved in cadherin stabilization at contact sites. Finally, we show that Rab35 regulates myoblast fusion, a major cellular process under the control of cadherin-dependent signaling. Taken together, these results reveal that Rab35 regulates cadherin-dependent AJ formation and myoblast fusion. PMID:23197472

  12. Rab5-mediated VE-cadherin internalization regulates the barrier function of the lung microvascular endothelium

    PubMed Central

    Yang, Junjun; Yao, Wei; Qian, Guisheng; Wei, Zhenghua

    2016-01-01

    The small GTPase Rab5 has been well defined to control the vesicle-mediated plasma membrane protein transport to the endosomal compartment. However, its function in the internalization of vascular endothelial (VE)-cadherin, an important component of adherens junctions, and as a result regulating the endothelial cell polarity and barrier function remain unknown. Here, we demonstrated that lipopolysaccharide (LPS) simulation markedly enhanced the activation and expression of Rab5 in human pulmonary microvascular endothelial cells (HPMECs), which is accompanied by VE-cadherin internalization. In parallel, LPS challenge also induced abnormal cell polarity and dysfunction of the endothelial barrier in HPMECs. LPS stimulation promoted the translocation of VE-cadherin from the plasma membrane to intracellular compartments, and intracellularly expressed VE-cadherin was extensively colocalized with Rab5. Small interfering RNA (siRNA)-mediated depletion of Rab5a expression attenuated the disruption of LPS-induced internalization of VE-cadherin and the disorder of cell polarity. Furthermore, knockdown of Rab5 inhibited the vascular endothelial hyperpermeability and protected endothelial barrier function from LPS injury, both in vitro and in vivo. These results suggest that Rab5 is a critical mediator of LPS-induced endothelial barrier dysfunction, which is likely mediated through regulating VE-cadherin internalization. These findings provide evidence, implicating that Rab5a is a potential therapeutic target for preventing endothelial barrier disruption and vascular inflammation. PMID:26112597

  13. Rab5-mediated VE-cadherin internalization regulates the barrier function of the lung microvascular endothelium.

    PubMed

    Yang, Junjun; Yao, Wei; Qian, Guisheng; Wei, Zhenghua; Wu, Guangyu; Wang, Guansong

    2015-12-01

    The small GTPase Rab5 has been well defined to control the vesicle-mediated plasma membrane protein transport to the endosomal compartment. However, its function in the internalization of vascular endothelial (VE)-cadherin, an important component of adherens junctions, and as a result regulating the endothelial cell polarity and barrier function remain unknown. Here, we demonstrated that lipopolysaccharide (LPS) simulation markedly enhanced the activation and expression of Rab5 in human pulmonary microvascular endothelial cells (HPMECs), which is accompanied by VE-cadherin internalization. In parallel, LPS challenge also induced abnormal cell polarity and dysfunction of the endothelial barrier in HPMECs. LPS stimulation promoted the translocation of VE-cadherin from the plasma membrane to intracellular compartments, and intracellularly expressed VE-cadherin was extensively colocalized with Rab5. Small interfering RNA (siRNA)-mediated depletion of Rab5a expression attenuated the disruption of LPS-induced internalization of VE-cadherin and the disorder of cell polarity. Furthermore, knockdown of Rab5 inhibited the vascular endothelial hyperpermeability and protected endothelial barrier function from LPS injury, both in vitro and in vivo. These results suggest that Rab5 is a critical mediator of LPS-induced endothelial barrier dysfunction, which is likely mediated through regulating VE-cadherin internalization. These findings provide evidence, implicating that Rab5a is a potential therapeutic target for preventing endothelial barrier disruption and vascular inflammation.

  14. Rab25 regulates invasion and metastasis in head and neck cancer

    PubMed Central

    Amornphimoltham, Panomwat; Rechache, Kamil; Thompson, Jamie; Masedunskas, Andrius; Leelahavanichkul, Kantima; Patel, Vyomesh; Molinolo, Alfredo; Gutkind, J. Silvio; Weigert, Roberto

    2013-01-01

    Purpose Head and neck squamous cell carcinoma (HNSCC) is one of the ten most common cancers with a 50% five-year survival rate, which has remained unchanged for the past three decades. One of the major reasons for the aggressiveness of this cancer is that HNSCCs readily metastasize to cervical lymph nodes that are abundant in the head and neck region. Hence, discovering new molecules controlling the metastatic process as well as understanding their regulation at the molecular level are essential for effective therapeutic strategies. Experimental Design Rab25 expression level was analyzed in HNSCC tissue microarray. We used a combination of intravital microscopy in live animals and immunofluorescence in an in vitro invasion assay, to study role of Rab25 in tumor cells migration and invasion. Results In this study, we identified the small GTPase Rab25 as a key regulator of HNSCC metastasis. We observed that Rab25 is downregulated in HNSCC patients. Next, we determined that re-expression of Rab25 in a metastatic cell line is sufficient to block invasion in a 3D collagen matrix and metastasis to cervical lymph nodes in a mouse model for oral cancer. Specifically, Rab25 affects the organization of F-actin at the cell surface, rather than cell proliferation, apoptosis or tumor angiogenesis. Conclusion These findings suggest that Rab25 plays an important role in tumor migration and metastasis, and that understanding its function may lead to the development of new strategies to prevent metastasis in oral cancer patients. PMID:23340300

  15. Purification and characterization of Ras related protein, Rab5a from Tinospora cordifolia.

    PubMed

    Amir, Mohd; Wahiduzzaman; Dar, Mohammad Aasif; Haque, Md Anzarul; Islam, Asimul; Ahmad, Faizan; Hassan, Md Imtaiyaz

    2016-01-01

    Ras related protein (Rab5a) is one of the most important member of the Rab family which regulates the early endosome fusion in endocytosis, and it also helps in the regulation of the budding process. Here, for the first time we report a simple and reproducible method for the purification of the Rab5a from a medicinal plant Tinospora cordifolia. We have used weak cation-exchange (CM-Sepharose-FF) followed by gel-filtration chromatography. A purified protein of 22-kDa was observed on SDS-PAGE which was identified as Rab5a using MALDI-TOF/MS. Our purification procedure is fast and simple with high yield. The purified protein was characterized using circular dichroism for the measurement of secondary structure followed by GdmCl- and urea-induced denaturation to calculate the values of Gibbs free energy change (ΔGD), ΔGD°, midpoint of the denaturation Cm, i.e. molar GdmCl [GdmCl] and molar urea [Urea] concentration at which ΔGD=0; and m, the slope (=∂ΔGD/∂[d]) values. Furthermore, thermodynamic properties of Rab5a were also measured by differential scanning calorimeter. Here, using isothermal calorimeteric measurements we further showed that Rab5a binds with the GTP. This is a first report on the purification and biophysical characterization of Rab5a protein from T. cordifolia.

  16. Distinct Sets of Rab6 Effectors Contribute to ZW10- and COG-Dependent Golgi Homeostasis

    PubMed Central

    Majeed, Waqar; Liu, Shijie; Storrie, Brian

    2014-01-01

    The organization of the Golgi apparatus is determined in part by the interaction of Rab proteins and their diverse array of effectors. Here, we used multiple approaches to identify and characterize a small subset of effectors that mimicked the effects of Rab6 on Golgi ribbon organization. In a visual-based, candidate-protein screen, we found that the individual depletion of any of three Rab6 effectors, myosin IIA (MyoIIA), Kif20A, and Bicaudal D (BicD), was sufficient to suppress Golgi ribbon fragmentation/dispersal coupled to retrograde tether proteins in a manner paralleling Rab6. MyoIIA and Kif20A depletion were pathway selective and suppressed ZW10-dependent Golgi ribbon fragmentation/dispersal only while BicD depletion, like Rab6, suppressed both ZW10- and COG-dependent Golgi ribbon fragmentation. The MyoIIA effects could be produced in short term assays by the reversible myosin inhibitor, blebbistatin. At the electron microscope level, the effects of BicD-depletion mimicked many of those of Rab6-depletion: longer and more continuous Golgi cisternae and a pronounced accumulation of coated vesicles. Functionally, BicD-depleted cells were inhibited in transport of newly synthesized VSV-G protein to the cell surface. In sum, our results indicate small, partially overlapping subsets of Rab6 effectors are differentially important to two tether-dependent pathways essential to Golgi organization and function. PMID:24575842

  17. Active Metal Brazing and Characterization of Brazed Joints in C-C and C-SiC Composites to Copper-Clad-Molybdenum System

    NASA Technical Reports Server (NTRS)

    Singh, M.; Asthana, R.

    2008-01-01

    Carbon/carbon composites with CVI and resin-derived matrices, and C/SiC composites reinforced with T-300 carbon fibers in a CVI SiC matrix were joined to Cu-clad Mo using two Ag-Cu braze alloys, Cusil-ABA (1.75% Ti) and Ticusil (4.5% Ti). The brazed joints revealed good interfacial bonding, preferential precipitation of Ti at the composite/braze interface, and a tendency toward delamination in resin-derived C/C composite. Extensive braze penetration of the inter-fiber channels in the CVI C/C composites was observed. The Knoop microhardness (HK) distribution across the C/C joints indicated sharp gradients at the interface, and a higher hardness in Ticusil than in Cusil-ABA. For the C/SiC composite to Cu-clad-Mo joints, the effect of composite surface preparation revealed that ground samples did not crack whereas unground samples cracked. Calculated strain energy in brazed joints in both systems is comparable to the strain energy in a number of other ceramic/metal systems. Theoretical predictions of the effective thermal resistance suggest that such joined systems may be promising for thermal management applications.

  18. Disease Mutations in Rab7 Result in Unregulated Nucleotide Exchange and Inappropriate Activation

    SciTech Connect

    B McCray; E Skordalakes; J Taylor

    2011-12-31

    Rab GTPases are molecular switches that orchestrate vesicular trafficking, maturation and fusion by cycling between an active, GTP-bound form, and an inactive, GDP-bound form. The activity cycle is coupled to GTP hydrolysis and is tightly controlled by regulatory proteins. Missense mutations of the GTPase Rab7 cause a dominantly inherited axonal degeneration known as Charcot-Marie-Tooth type 2B through an unknown mechanism. We present the 2.8 A crystal structure of GTP-bound L129F mutant Rab7 which reveals normal conformations of the effector binding regions and catalytic site, but an alteration to the nucleotide binding pocket that is predicted to alter GTP binding. Through extensive biochemical analysis, we demonstrate that disease-associated mutations in Rab7 do not lead to an intrinsic GTPase defect, but permit unregulated nucleotide exchange leading to both excessive activation and hydrolysis-independent inactivation. Consistent with augmented activity, mutant Rab7 shows significantly enhanced interaction with a subset of effector proteins. In addition, dynamic imaging demonstrates that mutant Rab7 is abnormally retained on target membranes. However, we show that the increased activation of mutant Rab7 is counterbalanced by unregulated, GTP hydrolysis-independent membrane cycling. Notably, disease mutations are able to rescue the membrane cycling of a GTPase-deficient mutant. Thus, we demonstrate that disease mutations uncouple Rab7 from the spatial and temporal control normally imposed by regulatory proteins and cause disease not by a gain of novel toxic function, but by misregulation of native Rab7 activity.

  19. Brazing of zirconia to Ti and Ti6Al4V

    SciTech Connect

    Agathopoulos, S.; Moretto, P.; Peteves, S.D.

    1997-11-01

    The interface microstructure of tetragonal zirconia (Y{sub 2}O{sub 3}-stab.), (TZP), brazed to itself, Ti and Ti6Al4V with a commercial Ag35Cu1.65Ti filler alloy at 900{degrees}C in vacuum was investigated. TiO formed in the reaction zone in TZP/TZP joints irrespectively the brazing time. In TZP/Ti joints, a complex interface microstructure comprising Ti{sub 2}O, Cu{sub 2}Ti{sub 4}O and Cu-Ti-intermetallics was time dependent. High joint strengths of over 400 MPa were obtained for short brazing times. In TZP/Ti6Al4V joints, the evolution of the microstructure with brazing time depends on diffusion of Ti, Al and V to the interface forming Cu{sub 2}(Ti,A1,V){sub 4}O.

  20. Effects of high temperature brazing and thermal cycling on mechanical properties of Hastelloy X.

    NASA Technical Reports Server (NTRS)

    Dicus, D. L.; Buckley, J. D.

    1973-01-01

    Data are presented on the effects of brazing alloy, brazing operation, thermal cycling, and combinations of these on the yield strength, elongation, ultimate tensile strength, and fatigue life of thin gage Hastelloy X. These data show that brazing with a Ni-Pd-Au alloy at 1461 K resulted in reductions of 35 percent in yield strength and elongation, 6 percent in ultimate tensile strength, and 18 percent in fatigue limit of Hastelloy X, as compared with as-received material. Subsequent exposure to 200 thermal cycles between 533 K and 1144 K after brazing caused further losses of 4 percent in yield strength, 8 percent in ultimate tensile strength, and 6 percent in fatigue limit.

  1. Tensile tests and metallography of brazed AISI 316L specimens after irradiation

    NASA Astrophysics Data System (ADS)

    Groot, P.; Franconi, E.

    1994-08-01

    Stainless steel type 316L tensile specimens were vacuum brazed with three kinds of alloys: BNi-5, BNi-6, and BNi-7. The specimens were irradiated up to 0.7 dpa at 353 K in the High Flux Reactor at JRC Petten, the Netherlands. Tensile tests were performed at a constant displacement rate of 10 -3 s -1 at room temperature in the ECN hot cell facility. BNi-5 brazed specimens showed ductile behaviour. Necking and fractures were localized in the plate material. BNi-6 and BNi-7 brazed specimens failed brittle in the brazed zone. This was preceded by uniform deformation of the plate material. Tensile test results of irradiated specimens showed higher stresses due to radiation hardening and a reduction of the elongation of the plate material compared to the reference. SEM examination of the irradiated BNi-6 and BNi-7 fracture surfaces showed nonmetallic phases. These phases were not found in the reference specimens.

  2. Stress level evaluation of thin films under thermal loading from a brazing process

    SciTech Connect

    Wang, Z.; Kuzay, T.M.

    1996-06-01

    X-ray front end beamline windows are made of thin beryllium foil that is commonly brazed or diffusion bonded onto a copper frame. In the brazing process, due to differences in the thermal expansion coefficient of the beryllium and copper materials, the beryllium film ends up in a state of very high level compression stress after cooling from 700{degrees}C (the brazing temperature) to room temperature. This makes the thin Be foil deform into a dome-shaped structure due to the usual asymmetrical geometry of the window. This paper studies the brazing process using a finite element method and explains the reason for such phenomena. Recommendations are offered for possible improvement in the bonding process of beryllium to the window block.

  3. Compatibility of Au-Cu-Ni braze alloy with NH3

    NASA Technical Reports Server (NTRS)

    Diaz, V., Jr.

    1978-01-01

    Tests show that Gold-Copper-Nickel alloy is compatible with ammonia systems. Joining tubes by brazing has advantages such as reducing chances of excessive grain growth in base metal, saving weight, and cleanliness.

  4. Fluxless Brazing and Heat Treatment of a Plate-Fin Sandwich Actively Cooled Panel

    NASA Technical Reports Server (NTRS)

    Beuyukian, C. S.

    1978-01-01

    The processes and techniques used to fabricate plate-fin sandwich actively cooled panels are presented. The materials were 6061 aluminum alloy and brazing sheet having clad brazing alloy. The panels consisted of small scale specimens, fatigue specimens, and a large 0.61 m by 1.22 m test panel. All panels were fluxless brazed in retorts in heated platen presses while exerting external pressure to assure intimate contact of details. Distortion and damage normally associated with that heat treatment were minimized by heat treating without fixtures and solution quenching in an organic polymer solution. The test panel is the largest fluxless brazed and heat treated panel of its configuration known to exist.

  5. Microstructural development of diffusion-brazed austenitic stainless steel to magnesium alloy using a nickel interlayer

    SciTech Connect

    Elthalabawy, Waled M.; Khan, Tahir I.

    2010-07-15

    The differences in physical and metallurgical properties of stainless steels and magnesium alloys make them difficult to join using conventional fusion welding processes. Therefore, the diffusion brazing of 316L steel to magnesium alloy (AZ31) was performed using a double stage bonding process. To join these dissimilar alloys, the solid-state diffusion bonding of 316L steel to a Ni interlayer was carried out at 900 deg. C followed by diffusion brazing to AZ31 at 510 deg. C. Metallographic and compositional analyses show that a metallurgical bond was achieved with a shear strength of 54 MPa. However, during the diffusion brazing stage B{sub 2} intermetallic compounds form within the joint and these intermetallics are pushed ahead of the solid/liquid interface during isothermal solidification of the joint. These intermetallics had a detrimental effect on joint strengths when the joint was held at the diffusion brazing temperature for longer than 20 min.

  6. Studies and research on the microstructure of brazed aluminum alloys in the repair process

    NASA Astrophysics Data System (ADS)

    Dimitescu, A.; Amza, Gh; Niţoi, D. F.; Amza, C. Gh; Apostolescu, Z.

    2016-08-01

    In aeronautical industry, in recent years, brazing joints got a growing spread. Therefore, it is necessary a detailed study on the microstructure of the assembly base and filler material brazed in conditions of reconditioning operation [1]. The methods of destructive examination are not associated with any particular type of test piece but lay down the general principles of the types of testing described. It is emphasized that a satisfactory examination method can only be developed and used after taking into account all the relevant factors regarding the equipment to be used and the characteristics of the test pieces being examined. [1, 2]. Brazing joints are most often systematically designed to be resistant to shearing, and the size of the joint influences the shear strength more than the tensile strength. Studies and researches on the microstructure may be necessary to determine the effects of brazing process or of any subsequent heat treatment on the characteristics of the joint.

  7. Method of fluxless brazing and diffusion bonding of aluminum containing components

    NASA Technical Reports Server (NTRS)

    Featherston, A. B.; Okelly, K. P. (Inventor)

    1976-01-01

    A method of diffusion bonding and fluxless brazing of aluminum containing components is reported. The aluminum surfaces are freed of any aluminum oxide coating and are coated with a polymeric sealer which can be thermally removed leaving essentially no residue. The polymeric sealer is being removed in a substantially oxygen free environment, and the aluminum components are then being brazed or diffusion bonded without the use of a flux to remove oxide coating.

  8. Flight/ground sample comparison relating to flight experiment M552, exothermic brazing

    NASA Technical Reports Server (NTRS)

    Heine, R. W.; Adams, C. M.; Siewert, T. A.

    1973-01-01

    Comparisons were made between Skylab and ground-based specimens of nickel and stainless steel which were vacuum brazed using silver-copper-lithium alloy with various joint configurations. It was established that the absence of gravity greatly extends the scope of brazing since capillary flow can proceed without gravity interference. There was also evidence of enhanced transport, primarily in that liquid silver copper alloy dissolves nickel to a much greater extent in the zero gravity environment.

  9. Some possible filler alloys with low vapor pressures for refractory-metal brazing

    NASA Technical Reports Server (NTRS)

    Morris, J. F.

    1973-01-01

    A compilation of eutectics and melting-point minima for binary combinations of metals having vapor pressures below 10 to the minus 10th power torr at 1500 degrees K and .00005 torr at 2000 degree K is presented. These compositions and others near them on their phase diagrams are potential special brazing fillers for refractory metals. Some possible problems and advantages for fusion bonds of such mixtures are indicated. Evaluations of brazing fillers containing refractory metals are reported.

  10. A technique for brazing graphite/graphite and stainless steel/high-carbon steel joints

    SciTech Connect

    Ohmura, H.; Kawashiri, K. . Dept. of Metals and Inorganic Materials); Yoshida, T. . Vehicle Machine Engineering Dept.); Yoshimoto, O. . Quality Control Dept.)

    1994-10-01

    A new brazing technique for joining graphite to itself or to metals such as Mo, W, or Cu, with conventional brazing filler metals has been developed. Essentially, it is impossible to braze graphite with Cu filler metal because no wetting occurs. However, when a graphite base material is combined with an Fe base metal in Cu brazing, the Fe base metal dissolves in molten Cu. Simultaneously, the dissolved Fe grows as part of a columnar Fe-9 [approximately] 6Cu-1.6C alloy phase at the graphite interface at a constant brazing temperature, that is, the dissolution and deposit of base metal takes place. By placing an Fe foil insert between both graphite base materials, therefore, the columnar phase is formed at both graphite faces and grows toward the Fe foil during heating. As a result, both graphite base materials are united by the columnar phase through the Fe foil. In the same way, a graphite/Mo or /W joint can be produced. Moreover, when using BAu-1, which has a lower melting point than that of BCu-1, it is also possible to braze graphite to Cu. The shear strength of a graphite/graphite joint with a 0.12-mm thick Fe foil at room temperature was about 32 MPa. Further, the bending strength of the graphite/graphite and /Cu joints at 873 K (1,112 F), as measured using the four-point bending test, was 35 and 11 MPa, respectively. In addition, the technique can be applied to the brazing of AISI 304 stainless steel to high-C steel with BCu-1 where, normally, Cr[sub 23]C[sub 6] and Cr[sub 7]C[sub 3] layers are formed at the high-C steel braze interface; these carbide layers result in the loss of mechanical properties of the joint.

  11. [Determination of Ag, Cu, Zn and Cd in silver brazing filler metals by ICP-AES].

    PubMed

    Yang, X

    1997-06-01

    A method of simultaneous and direct determination for Ag, Cu, Zn and Cd in silver brazing filler metals by ICP-AES is reported. The spectral interferences and effect of acidity have been investigated. Working conditions were optimized. The method has been applied to the analysis of silver brazing filler metals with RSD of 4-7% and recovery of 94-105%. This method was accurate, simple and rapid.

  12. Study of the possibility of using solar radiant energy for welding and brazing metals

    NASA Technical Reports Server (NTRS)

    Dvernyakov, V. S.; Frantsevich, I. N.; Pasichnyy, V. V.; Shiganov, N. A.; Korunov, Y. I.; Kasich-Pilipenko, I. Y.

    1974-01-01

    The solar spectrum at the surface of the earth is analyzed. A facility for creating concentrated solar radiant energy flux is described, and data on its energetic capabilities are presented. The technology of solar welding by the fusion technique and joining by high-temperature brazing is examined. The use of concentrated solar radiant energy for welding and brazing metals and alloys is shown. The results of mechanical tests and microscopic and macroscopic studies are presented.

  13. Development of plasma MIG brazing process for dissimilar metal joining of aluminum to steel

    NASA Astrophysics Data System (ADS)

    Tashiro, Shinichi; Tanaka, Manabu

    2014-08-01

    This study aims to develop a new brazing process employing plasma MIG. Because the energy density of the plasma produced by the plasma electrode is low, the base metal can be heated extensively without melting of the base metal, consequently improving the wettability of bead. This paper discussed the dissimilar metal joining of aluminum to steel by plasma MIG brazing process. Fracture occurred at the HAZ in the aluminum plate at 80 MPa.

  14. Mechanical properties of Inconel 718 and Nickel 201 alloys after thermal histories simulating brazing and high temperature service

    NASA Technical Reports Server (NTRS)

    James, W. F.

    1985-01-01

    An experimental investigation was made to evaluate two nickel base alloys (Nickel-201 and Inconel-718) in three heat treated conditions. These conditions were: (1) annealed; (2) after thermal exposure simulating a braze cycle; and (3) after a thermal exposure simulating a braze cycle plus one operational lifetime of high temperature service. For the Nickel-201, two different braze cycle temperatures were evaluated. A braze cycle utilizing a lower braze temperature resulted in less grain growth for Nickel-201 than the standard braze cycle used for joining Nickel-201 to Inconel-718. It was determined, however, that Nickel-201, was marginal for temperatures investigated due to large grain growth. After the thermal exposures described above, the mechanical properties of Nickel-201 were degraded, whereas similar exposure on Inconel-718 actually strengthened the material compared with the annealed condition. The investigation included tensile tests at both room temperature and elevated temperatures, stress-rupture tests, and metallographic examination.

  15. Braze alloy process and strength characterization studies for 18 nickel grade 200 maraging steel with application to wind tunnel models

    NASA Technical Reports Server (NTRS)

    Bradshaw, James F.; Sandefur, Paul G., Jr.; Young, Clarence P., Jr.

    1991-01-01

    A comprehensive study of braze alloy selection process and strength characterization with application to wind tunnel models is presented. The applications for this study include the installation of stainless steel pressure tubing in model airfoil sections make of 18 Ni 200 grade maraging steel and the joining of wing structural components by brazing. Acceptable braze alloys for these applications are identified along with process, thermal braze cycle data, and thermal management procedures. Shear specimens are used to evaluate comparative shear strength properties for the various alloys at both room and cryogenic (-300 F) temperatures and include the effects of electroless nickel plating. Nickel plating was found to significantly enhance both the wetability and strength properties for the various braze alloys studied. The data are provided for use in selecting braze alloys for use with 18 Ni grade 200 steel in the design of wind tunnel models to be tested in an ambient or cryogenic environment.

  16. Tensile fracture characterization of braze joined copper-to-CFC coupon assemblies

    NASA Astrophysics Data System (ADS)

    Trester, P. W.; Valentine, P. G.; Johnson, W. R.; Chin, E.; Reis, E. E.; Colleraine, A. P.

    1996-10-01

    A vacuum brazing process was used to join a broad spectrum of carbon-fiber reinforced carbon matrix composite (CFC) materials, machined into cylindrical coupons, between coupons of oxygen-free copper, the braze alloy was a copper-base alloy which contained only low activation elements (Al, Si, and Ti) relative to a titanium baseline specification. This demonstration was of particular importance for plasma facing components (PFCs) under design for use in the Tokamak Physics Experiment (TPX); the braze investigation was conducted by General Atomics for the Princeton Plasma Physics Laboratory. A tensile test of each brazed assembly was conducted. The results from the braze processing, testing, and fracture characterization studies of this reporting support the use of CFC's of varied fiber architecture and matrix processing in PFC designs for TPX. Further, the copper braze alloy investigated is now considered to be a viable candidate for a low-activation bond design. The prediction of plasma disruption-induced loads on the PFCs in TPX requires that joint strength between CFC tiles and their copper substrate be considered in design analysis and CFC selection.

  17. Chemical elements diffusion in the stainless steel components brazed with Cu-Ag alloy

    NASA Astrophysics Data System (ADS)

    Voiculescu, I.; Geanta, V.; Vasile, I. M.; Binchiciu, E. F.; Winestoock, R.

    2016-06-01

    The paper presents the study of diffusion of chemical elements through a brazing joint, between two thin components (0.5mm) made of stainless steel 304. An experimental brazing filler material has been used for brazing stainless steel component and then the diffusion phenomenon has been studied, in terms of chemical element displacement from the brazed separation interface. The filler material is in the form of a metal rod coated with ceramic slurry mixture of minerals, containing precursors and metallic powders, which can contribute to the formation of deposit brazed. In determining the distance of diffusion of chemical elements, on both sides of the fusion line, were performed measurements of the chemical composition using electron microscopy SEM and EDX spectrometry. Metallographic analysis of cross sections was performed with the aim of highlight the microstructural characteristics of brazed joints, for estimate the wetting capacity, adherence of filler metal and highlight any imperfections. Analyzes performed showed the penetration of alloying elements from the solder (Ag, Cu, Zn and Sn) towards the base material (stainless steel), over distances up to 60 microns.

  18. Analysis of Diffusional Solidification in a Wide-Gap Brazing Powder Mixture Using Differential Scanning Calorimetry

    NASA Astrophysics Data System (ADS)

    Corbin, Stephen F.; Murray, D. Clark; Bouthillier, Alain

    2016-12-01

    The diffusional solidification (DS) of a mixed powder system, consisting of pure Ni base metal powder and BNi-2 braze powder, suitable for wide-gap brazing was investigated using differential scanning calorimetry (DSC) and parallel microstructural examination. It was determined that very little interdiffusion between the powders developed in the solid state prior to braze powder melting. Once liquid formed, rapid DS occurred such that, when the powders were loosely mixed together, only 20 to 50 wt pct of the potential liquid fraction actually developed, leading to poor densification. Separating the braze and Ni into a layered structure leads to less DS, increased liquid formation ( i.e., 35 to 80 wt pct of the potential liquid fraction) and improved densification. The rate of isothermal solidification in layered structures consisting of 30 and 40 wt pct BNi-2 braze material was determined using DSC. After 30 minutes of braze time at 1323 K (1050 °C), complete IS occurred, thus avoiding the formation of a continuous network of large borides. The final microstructure of the IS mixtures consisted of a continuous matrix of solid solution Ni, with isolated and dispersed borides.

  19. Laser brazing of inconel 718 alloy with a silver based filler metal

    NASA Astrophysics Data System (ADS)

    Khorram, A.; Ghoreishi, M.; Torkamany, M. J.; Bali, M. M.

    2014-03-01

    In the presented study laser brazing of an inconel 718 alloy with silver based filler metal using 400 W pulsed Nd:YAG laser is investigated. Laser brazing was performed with varying laser frequency, pulse width, process speed and gap distance. The effect of preheating on wetting and spreading also was studied. Brazing geometrical images were observed using an optical microscope. The composition analysis and microstructure of the filler metal and brazed joints were examined using X-ray diffraction analyzer (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS). Micro-hardness and tensile test were performed for investigation of mechanical properties. The experimental observations show that filler metal consist of α-Ag solid solution, ά-Cu solid solution surround by the α-Ag solid solution and eutectic structure. Phases of the brazed joint are similar to the filler metal. The results indicate that the filler metal has adequate wetting and spreading on inconel 718 and the wetting angle depends on the heat input significantly. Interdiffusion occurs in laser brazing and the average thickness of reaction layer is approximately 2.5 μm. Whenever the gap is big, it is needed to use longer pulse width in order to have a better melting flow. Preheating has significant influence on wetting and spreading of the filler metal.

  20. Brazed graphite/refractory metal composites for first-wall protection elements

    NASA Astrophysics Data System (ADS)

    Šmid, I.; Croessmann, C. D.; Salmonson, J. C.; Whitley, J. B.; Kny, E.; Reheis, N.; Kneringer, G.; Nickel, H.

    1991-03-01

    The peak surface heat flux deposition on divertor elements of near term fusion devices is expected to exceed 10 MW/m 2. The needed reliability of brazed plasma interactive components, particularly under abnormal operating conditions with peak surface temperatures well beyond 1000°C, makes refractory metallic substrates and brazes with a high melting point very attractive. TZM, a high temperature alloy of molybdenum, and isotropic graphite, materials very closely matched in their thermal expansion, were brazed with four high-temperature brazes. The brazes used were Zr, 90Ni/10Ti, 90Cu/10Ti and 70Ag/27Cu/3Ti (nominal composition prior to brazing, wt%). The resulting composite tiles of 50 × 50 mm2 with a TZM thickness of 5 mm and a graphite thickness of 10 mm have been tested in high heat flux simulation for their thermal fatigue properties. Up to 600 loading cycles were carried out with an average heat flux of 10 MW/m 2 for 0.5 s pulses. The maximum surface temperature was 1100°C. In support of the experiment, the thermal response and temperature gradients of the samples were investigated using a finite element model.