WorldWideScience

Sample records for purified tpc isoforms

  1. TPC1 has two variant isoforms, and their removal has different effects on endo-lysosomal functions compared to loss of TPC2.

    Science.gov (United States)

    Ruas, Margarida; Chuang, Kai-Ting; Davis, Lianne C; Al-Douri, Areej; Tynan, Patricia W; Tunn, Ruth; Teboul, Lydia; Galione, Antony; Parrington, John

    2014-11-01

    Organelle ion homeostasis within the endo-lysosomal system is critical for physiological functions. Two-pore channels (TPCs) are cation channels that reside in endo-lysosomal organelles, and overexpression results in endo-lysosomal trafficking defects. However, the impact of a lack of TPC expression on endo-lysosomal trafficking is unknown. Here, we characterize Tpcn1 expression in two transgenic mouse lines (Tpcn1(XG716) and Tpcn1(T159)) and show expression of a novel evolutionarily conserved Tpcn1B transcript from an alternative promoter, raising important questions regarding the status of Tpcn1 expression in mice recently described to be Tpcn1 knockouts. We show that the transgenic Tpcn1(T159) line lacks expression of both Tpcn1 isoforms in all tissues analyzed. Using mouse embryonic fibroblasts (MEFs) from Tpcn1(-/-) and Tpcn2(-/-) animals, we show that a lack of Tpcn1 or Tpcn2 expression has no significant impact on resting endo-lysosomal pH or morphology. However, differential effects in endo-lysosomal function were observed upon the loss of Tpcn1 or Tpcn2 expression; thus, while Tpcn1(-/-) MEFs have impaired trafficking of cholera toxin from the plasma membrane to the Golgi apparatus, Tpcn2(-/-) MEFs show slower kinetics of ligand-induced platelet-derived growth factor receptor β (PDGFRβ) degradation, which is dependent on trafficking to lysosomes. Our findings indicate that TPC1 and TPC2 have important but distinct roles in the endo-lysosomal pathway.

  2. TPC1 Has Two Variant Isoforms, and Their Removal Has Different Effects on Endo-Lysosomal Functions Compared to Loss of TPC2

    OpenAIRE

    Ruas, Margarida; Chuang, Kai-Ting; Davis, Lianne C.; Al-Douri, Areej; Tynan, Patricia W.; Tunn, Ruth; Teboul, Lydia; Galione, Antony; Parrington, John

    2014-01-01

    Organelle ion homeostasis within the endo-lysosomal system is critical for physiological functions. Two-pore channels (TPCs) are cation channels that reside in endo-lysosomal organelles, and overexpression results in endo-lysosomal trafficking defects. However, the impact of a lack of TPC expression on endo-lysosomal trafficking is unknown. Here, we characterize Tpcn1 expression in two transgenic mouse lines (Tpcn1 XG716 and Tpcn1 T159) and show expression of a novel evolutionarily conserved ...

  3. Characterization of Affinity-Purified Isoforms of Acinetobacter calcoaceticus Y1 Glutathione Transferases

    Directory of Open Access Journals (Sweden)

    Chin-Soon Chee

    2014-01-01

    Full Text Available Glutathione transferases (GST were purified from locally isolated bacteria, Acinetobacter calcoaceticus Y1, by glutathione-affinity chromatography and anion exchange, and their substrate specificities were investigated. SDS-polyacrylamide gel electrophoresis revealed that the purified GST resolved into a single band with a molecular weight (MW of 23 kDa. 2-dimensional (2-D gel electrophoresis showed the presence of two isoforms, GST1 (pI 4.5 and GST2 (pI 6.2 with identical MW. GST1 was reactive towards ethacrynic acid, hydrogen peroxide, 1-chloro-2,4-dinitrobenzene, and trans,trans-hepta-2,4-dienal while GST2 was active towards all substrates except hydrogen peroxide. This demonstrated that GST1 possessed peroxidase activity which was absent in GST2. This study also showed that only GST2 was able to conjugate GSH to isoproturon, a herbicide. GST1 and GST2 were suggested to be similar to F0KLY9 (putative glutathione S-transferase and F0KKB0 (glutathione S-transferase III of Acinetobacter calcoaceticus strain PHEA-2, respectively.

  4. Characterization of affinity-purified isoforms of Acinetobacter calcoaceticus Y1 glutathione transferases.

    Science.gov (United States)

    Chee, Chin-Soon; Tan, Irene Kit-Ping; Alias, Zazali

    2014-01-01

    Glutathione transferases (GST) were purified from locally isolated bacteria, Acinetobacter calcoaceticus Y1, by glutathione-affinity chromatography and anion exchange, and their substrate specificities were investigated. SDS-polyacrylamide gel electrophoresis revealed that the purified GST resolved into a single band with a molecular weight (MW) of 23 kDa. 2-dimensional (2-D) gel electrophoresis showed the presence of two isoforms, GST1 (pI 4.5) and GST2 (pI 6.2) with identical MW. GST1 was reactive towards ethacrynic acid, hydrogen peroxide, 1-chloro-2,4-dinitrobenzene, and trans,trans-hepta-2,4-dienal while GST2 was active towards all substrates except hydrogen peroxide. This demonstrated that GST1 possessed peroxidase activity which was absent in GST2. This study also showed that only GST2 was able to conjugate GSH to isoproturon, a herbicide. GST1 and GST2 were suggested to be similar to F0KLY9 (putative glutathione S-transferase) and F0KKB0 (glutathione S-transferase III) of Acinetobacter calcoaceticus strain PHEA-2, respectively.

  5. Screening of L-histidine-based ligands to modify monolithic supports and selectively purify the supercoiled plasmid DNA isoform.

    Science.gov (United States)

    Amorim, Lúcia F A; Sousa, Fani; Queiroz, João A; Cruz, Carla; Sousa, Ângela

    2015-06-01

    The growing demand of pharmaceutical-grade plasmid DNA (pDNA) suitable for biotherapeutic applications fostered the development of new purification strategies. The surface plasmon resonance technique was employed for a fast binding screening of l-histidine and its derivatives, 1-benzyl-L-histidine and 1-methyl-L-histidine, as potential ligands for the biorecognition of three plasmids with different sizes (6.05, 8.70, and 14 kbp). The binding analysis was performed with different isoforms of each plasmid (supercoiled, open circular, and linear) separately. The results revealed that the overall affinity of plasmids to l-histidine and its derivatives was high (KD  > 10(-8)  M), and the highest affinity was found for human papillomavirus 16 E6/E7 (K(D)  = 1.1 × 10(-10)  M and KD  = 3.34 × 10(-10)  M for open circular and linear plasmid isoforms, respectively). L-Histidine and 1-benzyl-L-histidine were immobilized on monolithic matrices. Chromatographic studies of L-histidine and 1-benzyl-L-histidine monoliths were also performed with the aforementioned samples. In general, the supercoiled isoform had strong interactions with both supports. The separation of plasmid isoforms was achieved by decreasing the ammonium sulfate concentration in the eluent, in both supports, but a lower salt concentration was required in the 1-benzyl-L-histidine monolith because of stronger interactions promoted with pDNA. The efficiency of plasmid isoforms separation remained unchanged with flow rate variations. The binding capacity for pDNA achieved with the l-histidine monolith was 29-fold higher than that obtained with conventional L-histidine agarose. Overall, the combination of either L-histidine or its derivatives with monolithic supports can be a promising strategy to purify the supercoiled isoform from different plasmids with suitable purity degree for pharmaceutical applications.

  6. ALICE installs its TPC

    CERN Multimedia

    2007-01-01

    The ALICE time projection chamber has been transported to the experimental cavern. The handling of this extremely fragile detector was a long and delicate process. The lorry transporting the TPC took one hour to travel from the assembly hall to the access shaft...200 metres away.The TPC was lowered into the ALICE experimental cavern with extreme care. The gap between the structure and the shaft wall was only 10 centimetres! For ALICE the year started with a flurry of activity...but at a snail's pace. On 8 January, the day CERN reopened after the end-of-year break, teams from ALICE and the TS Department began the transportation of the experiment's time projection chamber (TPC), the largest ever built. This 5-metre long and 5-m diameter cylinder was transported from the clean room where it had been assembled to the experimental cavern. The 300-metre journey took no less than four days! Since the TPC is an extremely fragile object, the utmost precautions were exercised in its transportation. The TPC, which is d...

  7. The ALICE TPC

    CERN Document Server

    Garabatos, C

    2004-01-01

    We describe the ALICE TPC, with emphasis on the design features which are driven by the physics requirements of the detector. In particular, the gas choice and composition, Ne-CO/sub 2/ Ý90-10¿, as well as the unprecedentedly high gain for a TPC (2*10/sup 40/), are direct consequences of the expected performance in the high- multiplicity environment of heavy-ion collisions at the LHC. The characteristics of this mixture are discussed and a viable way of improving the stability of detectors working under these conditions, namely the addition of nitrogen into the mixture, is presented. This results in a more effective Penning transfer of neon excited states onto ionisation of the quencher at no penalty for the charge transport and amplification properties.

  8. ALICE TPC commissioning results

    Energy Technology Data Exchange (ETDEWEB)

    Larsen, D.T., E-mail: dagtl@ift.uib.n [Department of Physics and Technology, University of Bergen, Bergen (Norway)

    2010-05-21

    ALICE is a dedicated heavy-ion experiment at CERN LHC aiming to study the properties of the quark-gluon plasma. A lead-lead collision might produce several 10 00 new particles. Detailed study of the event requires precise measurements of the particle tracks. A 90m{sup 3} Time Projection Chamber (TPC) with more than 500 000 read-out pads was built as the main central barrel tracker. Collisions can be recorded at a rate of up to about 1 kHz. The front-end electronics, designed from FPGAs and custom ASICs, performs shaping, amplification, digitisation and digital filtering of the signals. The data are forwarded to DAQ via 216 1.25 Gb/s fibre-optical links. Configuration, control and monitoring is done by an embedded Linux system on the front-end electronics. Before production runs with beam, extensive commissioning using tracks from cosmics and from the laser system as well as clusters from radioactive krypton gas is needed. Extensive results have been obtained with respect to the performance of the TPC including its sub-systems.

  9. On TPC cluster reconstruction

    CERN Document Server

    Dydak, F; Nefedov, Y; Wotschack, J; Zhemchugov, A

    2004-01-01

    For a bias-free momentum measurement of TPC tracks, the correct determination of cluster positions is mandatory. We argue in particular that (i) the reconstruction of the entire longitudinal signal shape in view of longitudinal diffusion, electronic pulse shaping, and track inclination is important both for the polar angle reconstruction and for optimum r phi resolution; and that (ii) self-crosstalk of pad signals calls for special measures for the reconstruction of the z coordinate. The problem of 'shadow clusters' is resolved. Algorithms are presented for accepting clusters as 'good' clusters, and for the reconstruction of the r phi and z cluster coordinates, including provisions for 'bad' pads and pads next to sector boundaries, respectively.

  10. The Gap-Tpc

    Science.gov (United States)

    Rossi, B.; Anastasio, A.; Boiano, A.; Catalanotti, S.; Cocco, A. G.; Covone, G.; Di Meo, P.; Longo, G.; Vanzanella, A.; Walker, S.; Wang, H.; Wang, Y.; Fiorillo, G.

    2016-02-01

    Several experiments have been conducted worldwide, with the goal of observing low-energy nuclear recoils induced by WIMPs scattering off target nuclei in ultra-sensitive, low-background detectors. In the last few decades noble liquid detectors designed to search for dark matter in the form of WIMPs have been extremely successful in improving their sensitivities and setting the best limits. One of the crucial problems to be faced for the development of large size (multi ton-scale) liquid argon experiments is the lack of reliable and low background cryogenic PMTs: their intrinsic radioactivity, cost, and borderline performance at 87 K rule them out as a possible candidate for photosensors. We propose a brand new concept of liquid argon-based detector for direct dark matter search: the Geiger-mode Avalanche Photodiode Time Projection Chamber (GAP-TPC) optimized in terms of residual radioactivity of the photosensors, energy and spatial resolution, light and charge collection efficiency.

  11. The GAP-TPC

    CERN Document Server

    Rossi, B; Boiano, A; Catalanotti, S; Cocco, A G; Covone, G; Di Meo, P; Longo, G; Vanzanella, A; Walker, S; Wang, H; Wang, Y; Fiorillo, G

    2016-01-01

    Several experiments have been conducted worldwide, with the goal of observing low-energy nuclear recoils induced by WIMPs scattering off target nuclei in ultra-sensitive, low-background detectors. In the last few decades noble liquid detectors designed to search for dark matter in the form of WIMPs have been extremely successful in improving their sensitivities and setting the best limits. One of the crucial problems to be faced for the development of large size (multi ton-scale) liquid argon experiments is the lack of reliable and low background cryogenic PMTs: their intrinsic radioactivity, cost, and borderline performance at 87 K rule them out as a possible candidate for photosensors. We propose a brand new concept of liquid argon-based detector for direct dark matter search: the Geiger-mode Avalanche Photodiode Time Projection Chamber (GAP-TPC) optimized in terms of residual radioactivity of the photosensors, energy and spatial resolution, light and charge collection efficiency

  12. The PANDA GEM-based TPC prototype

    Energy Technology Data Exchange (ETDEWEB)

    Fabbietti, L., E-mail: laura.fabbietti@ph.tum.d [Excellence Cluster ' Universe' , Technische Universitaet Muenchen, Boltztmannstr. 2, Garching bei Muenchen 85748 (Germany); Angerer, H. [Physik Department E18, Technische Universitaet Muenchen, 85748 Muenchen (Germany); Arora, R. [Detektor Labor des GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, 64291 Darmstadt (Germany); Beck, R. [Helmholtz-Institut fuer Strahlen- und Kernphysik, 53115 Bonn (Germany); Berger, M. [Excellence Cluster ' Universe' , Technische Universitaet Muenchen, Boltztmannstr. 2, Garching bei Muenchen 85748 (Germany); Buehler, P.; Cargnelli, M. [Stefan-Meyer Institute fuer subatomare Physik, 1090 Vienna (Austria); Doerheim, S. [Physik Department E18, Technische Universitaet Muenchen, 85748 Muenchen (Germany); Hehner, J. [Detektor Labor des GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, 64291 Darmstadt (Germany); Herrmann, N. [FOPI Collaboration, GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, 64291 Darmstadt (Germany); Hoeppner, C. [Physik Department E18, Technische Universitaet Muenchen, 85748 Muenchen (Germany); Kaiser, D. [Helmholtz-Institut fuer Strahlen- und Kernphysik, 53115 Bonn (Germany); Ketzer, B. [Physik Department E18, Technische Universitaet Muenchen, 85748 Muenchen (Germany); Mladen, K. [FOPI Collaboration, GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, 64291 Darmstadt (Germany); Kleipa, V. [Detektor Labor des GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, 64291 Darmstadt (Germany); Konorov, I. [Physik Department E18, Technische Universitaet Muenchen, 85748 Muenchen (Germany); Kunkel, J. [Detektor Labor des GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, 64291 Darmstadt (Germany); Lalik, R. [Excellence Cluster ' Universe' , Technische Universitaet Muenchen, Boltztmannstr. 2, Garching bei Muenchen 85748 (Germany); Lang, M. [Helmholtz-Institut fuer Strahlen- und Kernphysik, 53115 Bonn (Germany)

    2011-02-01

    We report on the development of a GEM-based TPC detector prototype for the PANDA experiment. The design and requirements of this device will be illustrated, with particular emphasis on the properties of the recently tested GEM-detector, the characterization of the read-out electronics and the development of the tracking software that allows to evaluate the GEM-TPC data.

  13. Purifying Nanomaterials

    Science.gov (United States)

    Hung, Ching-Cheh (Inventor); Hurst, Janet (Inventor)

    2014-01-01

    A method of purifying a nanomaterial and the resultant purified nanomaterial in which a salt, such as ferric chloride, at or near its liquid phase temperature, is used to penetrate and wet the internal surfaces of a nanomaterial to dissolve impurities that may be present, for example, from processes used in the manufacture of the nanomaterial.

  14. First TPC-Energy Benchmark: Lessons Learned in Practice

    Science.gov (United States)

    Young, Erik; Cao, Paul; Nikolaiev, Mike

    TPC-Energy specification augments the existing TPC Benchmarks with Energy Metrics. The TPC-Energy specification is designed to help hardware buyers identify energy efficient equipment that meets both their computational and budgetary requirements. In this paper we discuss our experience is publishing industry's first-ever TPC-Energy metric publication.

  15. TPC track distortions III: fiat lux

    CERN Document Server

    Boyko, I; Dydak, F; Elagin, A; Gostkin, M; Guskov, A; Koreshev, V; Nefedov, Y; Nikolaev, K; Veenhof, R; Wotschack, J; Zhemchugov, A

    2005-01-01

    We present a comprehensive overview and final summary of all four types of static track distortions seen in the HARP TPC, in terms of physical origins, mathematical modelling, and correction algorithms. 'Static'™ distortions are defined as not depending on the event time within the 400 ms long accelerator spill. Calculated static distortions are compared with measurements from cosmic-muon tracks. We characterize track distortions by the r phi residuals of cluster positions with respect to the transverse projection of a helical trajectory constrained by hits in the RPC overlap regions. This method provides a fixed TPC-external reference system (by contrast to the co-moving coordinate system associated with a fit) which solely permits to identify individually, and measure quantitatively, the static TPC track distortions arising from (i) the inhomogeneity of the solenoidal magnetic field, (ii) the inhomogeneity of the electric field from the high-voltage mismatch between the inner and outer TPC field cages, (...

  16. Readout scheme of the upgraded ALICE TPC

    CERN Document Server

    Appelshaeuser, Harald; Ivanov, Marian; Lippmann, Christian; Wiechula, Jens

    2016-01-01

    In this document, we present the updated readout scheme for the ALICE TPC Upgrade. Two major design changes are implemented with respect to the concept that was presented in the TPC Upgrade Technical Design Report: – The SAMPA front-end ASIC will be used in direct readout mode. – The ADC sampling frequency will be reduced from 10 to 5 MHz. The main results from simulations and a description of the new readout scheme is outlined.

  17. Large bulk Micromegas detectors for TPC applications

    CERN Document Server

    Anvar, S; Boyer, M; Beucher, J; Calvet, D; Colas, P; De La Broise, X; Delagnes, E; Delbart, A; Druillole, F; Emery, S; Giganti, C; Giomataris, I; Mazzucato, E; Monmarthe, E; Nizery, F; Pierre, F; Ritou, J L; Sarrat, A; Zito, M; Catanesi, M G; Radicioni, E; De Oliveira, R; Blondel, A; Di Marco, M; Ferrere, D; Perrin, E; Ravonel, M; Jover, G; Lux, T; Rodriguez, A Y; Sanchez, F; Cervera, A; Hansen, C; Monfregola, L

    2009-01-01

    A large volume TPC will be used in the near future in a variety of experiments including T2K. The bulk Micromegas detector for this TPC is built using a novel production technique particularly suited for compact, thin and robust low mass detectors. The capability to pave a large surface with a simple mounting solution and small dead space is of particular interest for these applications. We have built several large bulk Micromegas detectors () and we have tested one in the former HARP field cage with a magnetic field. Prototypes cards of the T2K front end electronics, based on the AFTER ASIC chip, have been used in this TPC test for the first time. Cosmic ray data have been acquired in a variety of experimental conditions. Good detector performances, space point resolution and energy loss measurement have been achieved.

  18. TPC track distortions IV: post tenebras lux

    CERN Document Server

    Ammosov, V; Boyko, I; Chelkov, G; Dedovitch, D; Dydak, F; Elagin, A; Gostkin, M; Guskov, A; Koreshev, V; Krumshtein, Z; Nefedov, Y; Nikolaev, K; Wotschack, J; Zhemchugov, A

    2007-01-01

    We present a comprehensive discussion and summary of static and dynamic track distortions in the HARP TPC in terms of physical origin, mathematical modelling and correction algorithms. `Static' distortions are constant with time, while `dynamic' distortions are distortions that occur only during the 400 ms long accelerator spill. The measurement of dynamic distortions, their mathematical modelling and the correction algorithms build on our understanding of static distortions. In the course of corroborating the validity of our static distortion corrections, their reliability and precision was further improved. Dynamic TPC distortions originate dominantly from the `stalactite' effect: a column of positive-ion charge starts growing at the begin of the accelerator spill, and continues growing with nearly constant velocity out from the sense-wire plane into the active TPC volume. However, the `stalactite' effect is not able to describe the distortions that are present already at the start of the spill and which ha...

  19. TPC1 - SV Channels Gain Shape

    Institute of Scientific and Technical Information of China (English)

    Rainer Hedrich; Irene Marten

    2011-01-01

    T The most prominent ion channel localized in plant vacuoles is the slow activating SV type. Slow vacuolar (SV)channels were discovered by patch clamp studies as early as 1986. In the following two decades, numerous studies revealed that these calcium- and voltage-activated, nonselective cation channels are expressed in the vacuoles of all plants and every plant tissue. The voltage-dependent properties of the SV channel are susceptible to modulation by calcium, pH, redox state, as well as regulatory proteins. In Arabidopsis, the SV channel is encoded by the AtTPC1 gene, and even though its gene product represents the by far largest conductance of the vacuolar membrane, tpc1-loss-of-function mutants appeared not to be impaired in major physiological functions such as growth, development, and reproduction. In contrast, the fou2 gain-of-function point mutation D454N within TPC1 leads to a pronounced growth phenotype and increased synthesis of the stress hormone jasmonate. Since the TPC1 gene is present in all land plants, it likely encodes a very general function. In this review, we will discuss major SV channel properties and their impact on plant cell physiology.

  20. Estimation of Weighting Potential for a TPC

    CERN Document Server

    Mukhopadhyay, S; Veenhof, R

    2007-01-01

    In this work, we have computed the three dimensional weighting potential, field and pad response function (PRF) of typical time projection chambers (TPC) using a recently formulated and developed boundary element method solver, namely, the nearly exact BEM solver. A realistic geometry of the device is found to have significant influence on the estimation of signal generation.

  1. Performance study of the neutron-TPC

    Science.gov (United States)

    Huang, Meng; Li, Yulan; Niu, Libo; Deng, Zhi; Cheng, Xiaolei; He, Li; Zhang, Hongyan; Fu, Jianqiang; Yan, Yangyang; Cai, Yiming; Li, Yuanjing

    2017-02-01

    Fast neutron spectrometers will play an important role in the future of the nuclear industry and nuclear physics experiments, in tasks such as fast neutron reactor monitoring, thermo-nuclear fusion plasma diagnostics, nuclear reaction cross-section measurement, and special nuclear material detection. Recently, a new fast neutron spectrometer based on a GEM (Gas Electron Multiplier amplification)-TPC (Time Projection Chamber), named the neutron-TPC, has been under development at Tsinghua University. It is designed to have a high energy resolution, high detection efficiency, easy access to the medium material, an outstanding n/γ suppression ratio, and a wide range of applications. This paper presents the design, test, and experimental study of the neutron-TPC. Based on the experimental results, the energy resolution (FWHM) of the neutron-TPC can reach 15.7%, 10.3% and 7.0% with detection efficiency higher than 10‑5 for 1.2 MeV, 1.81 MeV and 2.5 MeV neutrons respectively. Supported by National Natural Science Foundation of China (11275109)

  2. ALICE Time Projection Chamber (TPC) Readout Sector in Lab

    CERN Multimedia

    2003-01-01

    The Time Projection Chamber (TPC) is the main particle tracking detector in ALICE. Charged particles crossing the gas of the TPC knock electrons out of their atoms, which drift in the eletric field. By measuring the arrival of electrons at the end of the chamber, at segments such as the one shown here, the TPC will reconstruct the paths of the original charged particles.

  3. Optimization of the resolution of a track drift chamber (TPC); Optimierung der Aufloesung einer Spurendriftkammer (TPC)

    Energy Technology Data Exchange (ETDEWEB)

    Schoenfelder, S.

    1993-09-01

    In the framework of this diploma thesis it was studied, how the position resolution of a TPC can be improved. For this first properties like electron drift velocity as well transverse and longitudinal diffusion of different gases in a test TPC were determined. Starting from the measurement results it is possible, to select chamber gases with suitable drift velocity and low diffusion and to improve by this the position resolution of a TPC. The second essential point of approach for the improvement of the position resolution of a TPC was the variation of the TPC geometry. Here the pad width, the gap distance, and the sampling frequency was varied and the influence on the resolution studied. The essential result hereby was the development of a TPC with a gap distance of 1 mm, which reduced the pad response width to 1.4 mm and by this contributed to a distinct improvement of the position resolution. In a last step a gas microstrip chamber with TPC pad readout was designed, in which the amplification wire planes of the TPC were replaced by a microstrip structure on a glass substrae. Hereby the proof-of-principle of such a facility was brought and first measurements on the position resolution and on the time behaviour of the gas amplification performed. Looking out to the future further measurements with this facility, like for example variation of the substrate material, the substrate thickness, or the manufacturing process of the microstrip structure for the improvement of the time stability of the gas amplification and the position resolution would be desirable. (HSI)

  4. Purified Humanism

    DEFF Research Database (Denmark)

    Nickelsen, Niels Christian Mossfeldt

    2016-01-01

    Abstract. The aim of the Leicester Conference is to help managers by way of experiential learning to acquire the prerequisites to influence effectively organizational change. For some time there has been an ongoing debate on the innovative potential of social psychological experiments...... and techniques. This article discusses the analytical possibilities of the notion “purified humanism” as part of an alternative analysis of the effective mechanisms of a widely used social psychological experiment. The article unfolds a number of ideas in relation to the socio-material provocations and maneuvers...

  5. Activity report of ILD-TPC Asia group

    CERN Document Server

    Kato, Y; Gros, P; Tian, J; Kawada, S; Fujii, K; Matsuda, T; Sugiyama, A; Nitoh, O; Watanabe, T; Fusayasu, T; Takahashi, T; Kobayashi, M

    2014-01-01

    The purpose of ILD-TPC Asia group is realization of high precision Time Projection Chamber (TPC) with Gas Electron Multiplier (GEM) as a central tracker in International Linear Collider (ILC). We have been studying the many R&D items to build the real detector as a member of LCTPC collaboration. This paper describes the our efforts for realization of the ILD-TPC, the result of test beam using large prototype TPC, local field distortion, positive ion effects and gate devices, and cooling electronics which are key items to build ILD-TPC.

  6. Purified humanism

    DEFF Research Database (Denmark)

    Nickelsen, Niels Christian Mossfeldt

    2016-01-01

    Abstract. The aim of the Leicester Conference is to help managers by way of experiential learning to acquire the prerequisites to influence effectively organizational change. For some time there has been an ongoing debate on the innovative potential of social psychological experiments...... and culturally specific attitudes in relation to leadership and the question of authority among participants. Keywords: The Leicester Conference, experiential learning, authority, socio-materiality, social techniques...... and techniques. This article discusses the analytical possibilities of the notion “purified humanism” as part of an alternative analysis of the effective mechanisms of a widely used social psychological experiment. The article unfolds a number of ideas in relation to the socio-material provocations and maneuvers...

  7. GEM Module Design for the ILD TPC

    CERN Document Server

    Benke, T; Münnich, A; Zenker, K

    2013-01-01

    A Time Projection Chamber (TPC) using micro-pattern gas detectors is planned as the main tracking device for a detector at the next Linear Collider. A novel support structure for Gas Electron Multipliers (GEMs), which minimizes the material and improves the flatness of the foils, has been developed and tested with multiple GEM modules in a large TPC prototype at DESY. Reducing dead material at the GEM module boundaries improves the field homogeneity. In addition, it was shown in simulation that a field shaping ring at the border of the module can improve the charge collection in regions with non-homogeneous fields. This shaping wire was integrated into the module design and a successful test beam campaign with three modules has been carried out. First results regarding resolution and field distortions will be discussed.

  8. Bulk micromegas detectors for large TPC applications

    CERN Document Server

    Bouchez, J; Cavata, Ch; Colas, P; De La Broise, X; Delbart, A; Giganon, Arnaud; Giomataris, Ioanis; Graffin, P; Mols, J Ph; Pierre, F; Ritou, J L; Sarrat, A; Virique, E; Zito, M; Radicioni, E; De Oliveira, R; Dumarchez, J; Abgrall, N; Bene, P; Blondel, A; Cervera-Villanueva, Anselmo; Ferrère, D; Maschiocchi, F; Perrin, E; Richeux, J P; Schroeter, R; Jover, G; Lux,; Rodriguez, A Y; Sánchez, F

    2007-01-01

    A large volume TPC will be used in the near future in a variety of experiments including T2K. The bulk Micromegas detector for this TPC is built using a novel production technique particularly suited for compact and robust low mass detectors. The capability to pave a large surface with a simple mounting solution and small dead space between modules is of particular interest for these applications. We have built several large bulk Micromegas detectors and we have tested them in the former HARP field cage setup with a magnetic field. Cosmic ray data have been acquired in a variety of experimental conditions. Good detector performances and space point resolution have been achieved.

  9. Strange particle measurements from the EOS TPC

    Energy Technology Data Exchange (ETDEWEB)

    Justice, M.

    1995-02-01

    A high statistics sample of {Lambda}`s produced in 2 GeV/nucleon {sup 5}8Ni + {sup nat}Cu collisions has been obtained with the EOS Time Projection Chamber at the Bevalac. The coverage of the EOS TPC is essentially 100% for y > y{sub cm} and extends down to P{sub T} = 0 where interesting effects such as collective radial expansion may be important. In addition, the detection of a majority of the charged particles in the TPC, along with the presence of directed flow for protons and heavier fragments at this beam energy, allows for the correlation of A production with respect to the event reaction plane. Our preliminary analysis indicates the first observation of a sidewards flow signature for A`s. Comparisons with the cascade code ARC are made.

  10. Highly integrated electronics for the star TPC

    Energy Technology Data Exchange (ETDEWEB)

    Arthur, A.A.; Bieser, F.; Hearn, W.; Kleinfelder, S.; Merrick, T.; Millaud, J.; Noggle, T.; Rai, G.; Ritter, H.G.; Wieman, H. [Lawrence Berkeley Laboratory, CA (United States)

    1991-12-31

    The concept for the STAR TPC front-end electronics is presented and the progress toward the development of a fully integrated solution is described. It is the goal of the R+D program to develop the complete electronics chain for the STAR central TPC detector at RHIC. It is obvious that solutions chosen e.g. for ALEPH are not adequate for the 150000 channels that need to be instrumented for readout. It will be necessary to perform all the signal processing, digitization and multiplexing directly on the detector in order to reduce per channel cost and the amount of cabling necessary to read out the information. We follow the approach chosen by the EOS TPC project, where the readout electronics on the detector consists of an integrated preamplifier, a hybrid shaping amplifier, an integrated switched capacitor array and a highly multiplexed ADC. The STAR electronics will be further integrated so that approximately 16 channels of the preamplifier, the shaper, the analog store and the ADC will be contained in two integrated circuits located directly on the pad plane.

  11. Momentum scale in the HARP TPC

    CERN Document Server

    Catanesi, M G; Edgecock, R; Ellis, M; Soler, F J P; Gössling, C; Bunyatov, S; Krasnoperov, A; Popov, B; Serdiouk, V; Tereschenko, V; Di Capua, E; Vidal-Sitjes, G; Artamonov, A; Giani, S; Gilardoni, S; Gorbunov, P; Grant, A; Grossheim, A; Ivanchenko, V; Kayis-Topaksu, A; Panman, J; Papadopoulos, I; Chernyaev, E; Tsukerman, I; Veenhof, R; Wiebusch, C; Zucchelli, P; Blondel, A; Borghi, S; Morone, M C; Prior, G; Schroeter, R; Meurer, C; Gastaldi, Ugo; Mills, G B; Graulich, J S; Grégoire, G; Bonesini, M; Ferri, F; Kirsanov, M; Bagulya, A; Grichine, V; Polukhina, N; Palladino, V; Coney, L; Schmitz, D; Barr, G; De Santo, A; Bobisut, F; Gibin, D; Guglielmi, A; Mezzetto, M; Dumarchez, J; Dore, U; Orestano, D; Pastore, F; Tonazzo, A; Tortora, L; Booth, C; Howlett, L; Bogomilov, M; Chizhov, M; Kolev, D; Tsenov, R; Piperov, S; Temnikov, P; Apollonio, M; Chimenti, P; Giannini, G; Burguet-Castell, J; Cervera-Villanueva, A; Gómez-Cadenas, J J; Martín-Albo, J; Novella, P; Sorel, M

    2007-01-01

    Recently a claim was made that the reconstruction of the large angle tracks in the HARP TPC was affected by a momentum bias as large as 15% at 500 MeV/c transverse momentum. In the following we recall the main issues with the momentum measurement in the HARP TPC, and describe the cross-checks made to validate the momentum scale. Proton-proton elastic scattering data off the hydrogen target are used to alibrate the momentum of charged particles with a precision evaluated to be 3.5%. A full description of the time development of the dynamic distortions in the TPC during physics spills is now available together with a correction algorithm. This allows a new cross-check using an enlarged data set made by comparing positive and negative pion elasticscattering data collected with negative polarity of the solenoid magnet. These data confirm the absence of a bias in the sagitta measurement. The dE/dx versus momentum curves are revisited, and shown to provide a confirmation that the HARP momentum calibration is correc...

  12. Highly integrated electronics for the star TPC

    Energy Technology Data Exchange (ETDEWEB)

    Arthur, A.A.; Bieser, F.; Hearn, W.; Kleinfelder, S.; Merrick, T.; Millaud, J.; Noggle, T.; Rai, G.; Ritter, H.G.; Wieman, H. [Lawrence Berkeley Laboratory, CA (United States)

    1991-12-31

    The concept for the STAR TPC front-end electronics is presented and the progress toward the development of a fully integrated solution is described. It is the goal of the R+D program to develop the complete electronics chain for the STAR central TPC detector at RHIC. It is obvious that solutions chosen e.g. for ALEPH are not adequate for the 150000 channels that need to be instrumented for readout. It will be necessary to perform all the signal processing, digitization and multiplexing directly on the detector in order to reduce per channel cost and the amount of cabling necessary to read out the information. We follow the approach chosen by the EOS TPC project, where the readout electronics on the detector consists of an integrated preamplifier, a hybrid shaping amplifier, an integrated switched capacitor array and a highly multiplexed ADC. The STAR electronics will be further integrated so that approximately 16 channels of the preamplifier, the shaper, the analog store and the ADC will be contained in two integrated circuits located directly on the pad plane.

  13. How to Advance TPC Benchmarks with Dependability Aspects

    Science.gov (United States)

    Almeida, Raquel; Poess, Meikel; Nambiar, Raghunath; Patil, Indira; Vieira, Marco

    Transactional systems are the core of the information systems of most organizations. Although there is general acknowledgement that failures in these systems often entail significant impact both on the proceeds and reputation of companies, the benchmarks developed and managed by the Transaction Processing Performance Council (TPC) still maintain their focus on reporting bare performance. Each TPC benchmark has to pass a list of dependability-related tests (to verify ACID properties), but not all benchmarks require measuring their performances. While TPC-E measures the recovery time of some system failures, TPC-H and TPC-C only require functional correctness of such recovery. Consequently, systems used in TPC benchmarks are tuned mostly for performance. In this paper we argue that nowadays systems should be tuned for a more comprehensive suite of dependability tests, and that a dependability metric should be part of TPC benchmark publications. The paper discusses WHY and HOW this can be achieved. Two approaches are introduced and discussed: augmenting each TPC benchmark in a customized way, by extending each specification individually; and pursuing a more unified approach, defining a generic specification that could be adjoined to any TPC benchmark.

  14. Inbetriebnahme und Kalibrierung der ALICE-TPC

    CERN Document Server

    Wiechula, Jens

    2008-01-01

    ALICE (A Large Ion Collider Experiment), is the dedicated heavy-ion experiment at the Large Hadron Collider (LHC) at CERN. It is optimised to reconstruct and identify the particles created in a lead-lead collision with a centre of mass energy of 5.5TeV. The main tracking detector is a large-volume time-projection chamber (TPC). With an active volume of about 88m^3 and a total readout area of 32.5m^2 it is the most challenging TPC ever build. A central electrode divides the 5m long detector into two drift regions. Each readout side is subdivided into 18 inner and 18 outer multi-wire proportional read-out chambers. The readout area is subdivide into 557568 pads, where each pad is read out by and electronics chanin. A complex calibration is needed in order to reach the design position-resolution of the reconstructed particle tracks of about 200um. One part of the calibration lies in understanding the electronic-response. The work at hand presents results of the pedestal and noise behaviour of the front-end elect...

  15. Team Primacy Concept (TPC) Based Employee Evaluation and Job Performance

    Science.gov (United States)

    Muniute, Eivina I.; Alfred, Mary V.

    2007-01-01

    This qualitative study explored how employees learn from Team Primacy Concept (TPC) based employee evaluation and how they use the feedback in performing their jobs. TPC based evaluation is a form of multirater evaluation, during which the employee's performance is discussed by one's peers in a face-to-face team setting. The study used Kolb's…

  16. CAMAC interface for TPC data-acquisition electronics

    Energy Technology Data Exchange (ETDEWEB)

    Sidman, S.; Olson, S.; Jared, R.

    1983-06-01

    The Time Projection Chamber (TPC) is a detector used for high-energy physics research at the Stanford PEP Accelerator. TPC requires about 17,000 channels of data acquisition, which samples on command the input to each channel at a 10 MHz rate. This high data rate is made possible by means of Charge Coupled Devices (CCDs), intelligent digitizers, and a sophisticated trigger system. The TPC-CAMAC interface described here was developed to allow experiments of smaller scale than the complete TPC to use the standard data acquisition portion of the TPC electronics, namely the amplifier, CCD and digitizer bins. These three bins, when properly interconnected and controlled by the interface control bin, form a transient digitizer with a depth of 455 samples and a maximum width of 256 channels per bin set.

  17. Isoforms of murine and human serum amyloid P component

    DEFF Research Database (Denmark)

    Nybo, Mads; Hackler, R; Kold, B

    1998-01-01

    Isoelectric focusing (IEF) and immunofixation of murine serum amyloid P component (SAP), purified and in serum, showed a distinct and strain-dependent isoform pattern with up to seven bands (pI 5.1-5.7). Neuraminidase treatment caused a shift of the isoforms to more basic pI values, but did...... of isoforms of human SAP required the presence of urea and higher SAP concentrations. TEF and immunofixation of SAP monomers showed five to eight isoforms, ranging from pI 4.7-5.7. IEF of SAP in human serum resulted in a less distinct pattern and more acidic isoforms. As with murine SAP, neuraminidase...

  18. Prototype endplate piece of the ALEPH Time Projection Chamber (TPC)

    CERN Multimedia

    1980-01-01

    Prototype piece of the ALEPH Time Projection Chamber (TPC), a large-volume tracking chamber. It is one of the endplates of a “Kind” sector, the smallest of the three types of sectors. The patterns etched into the copper form the cathode pads that measured particle track coordinates in the r-phi direction. The ALEPH TPC was the largest TPC at the time. It included a laser calibration system, a gating system to prevent space charge buildup, and a new radial pad geometry to improve resolution. the ALEPH TPC allowed for precise momentum measurements of the high-momentum particles from W and Z decays.The following institutes participated: CERN, Athens, Glasgow, Mainz, MPI Munich, INFN-Pisa, INFN-Trieste, Wisconsin.

  19. A radial TPC for heavy ions

    CERN Document Server

    Garabatos, C

    2000-01-01

    The CERES experiment at the CERN SPS has been recently upgraded with a TPC with radial drift field, the first one of its sort. Constructed during 1998, it has been successfully operated in commissioning and physics runs, with muon, proton, and heavy-ion beams. A high voltage electrode of about 0.5 m radius is surrounded by sixteen 2 m long readout chambers, placed at a radius of 1.3 m, with chevron-shaped readout pads. The field cage is enclosed by two low-mass voltage degraders at each end of the cylindrical structure. A Ne-CO/sub 2/ [80-20] gas mixture allows for a safe operation and good transport properties under drift fields ranging from 200 to 600 V/cm. A spatial resolution better than 700 microns and 350 microns in r and rdelta (phi), respectively, has been achieved in a highly inhomogeneous magnetic field. Details of its construction as well as results of the operation and performance in a high multiplicity environment are presented. (0 refs).

  20. Imaging in (high pressure) Micromegas TPC detectors

    Science.gov (United States)

    Luzón, G.; Cebrián, S.; Castel, J.; Dafni, Th.; Galán, J.; Garza, J. G.; Irastorza, I. G.; Iguaz, F. J.; Mirallas, H.; Ruíz-Choliz, E.

    2016-11-01

    The T-REX project of the group of the University of Zaragoza includes a number of R&D and prototyping activities to explore the applicability of gaseous Time Projection Chambers (TPCs) with Micromesh Gas Structures (Micromegas) in rare event searches where the pattern recognition of the signal is crucial for background discrimination. In the CAST experiment (CERN Axion Solar Telescope) a background level as low as 0.8 × 10-6 counts keV-1 cm-2 s-1 was achieved. Prototyping and simulations promise a 105 better signal-to-noise ratio than CAST for the future IAXO (International Axion Observatory) using x-ray telescopes. A new strategy is also explored in the search of WIMPS based on high gas pressure: the TREX-DM experiment, a low energy threshold detector. In both cases, axion and WIMP searches, the image of the expected signal is quite simple: a one cluster deposition coming from the magnet bore in the case of axions and, if possible, with a tadpole form in the case of WIMPs. It is the case of double beta decay (DBD) where imaging and pattern recognition play a major role. Results obtained in Xe + trimethylamine (TMA) mixture point to a reduction in electron diffusion which improves the quality of the topological pattern, with a positive impact on the discrimination capability, as shown in TREX-ββ prototype. Microbulk Micromegas are able to image the DBD ionization signature with high quality while, at the same time, measuring its energy deposition with a resolution of at least a ~ 3% FWHM at the transition energy Qββ and even better (up to ~ 1% FWHM) as extrapolated from low energy events. That makes Micromegas-based HPXe TPC a very competitive technique for the next generation DBD experiments (as PANDAX-III). Here, it will be shown the last results of the TREX project detectors and software concerning Axions, Dark matter and double beta decay.

  1. On the cellular site of two-pore channel TPC1 action in the Poaceae.

    Science.gov (United States)

    Dadacz-Narloch, Beata; Kimura, Sachie; Kurusu, Takamitsu; Farmer, Edward E; Becker, Dirk; Kuchitsu, Kazuyuki; Hedrich, Rainer

    2013-11-01

    The slow vacuolar (SV) channel has been characterized in different dicots by patch-clamp recordings. This channel represents the major cation conductance of the largest organelle in most plant cells. Studies with the tpc1-2 mutant of the model dicot plant Arabidopsis thaliana identified the SV channel as the product of the TPC1 gene. By contrast, research on rice and wheat TPC1 suggested that the monocot gene encodes a plasma membrane calcium-permeable channel. To explore the site of action of grass TPC1 channels, we expressed OsTPC1 from rice (Oryza sativa) and TaTPC1 from wheat (Triticum aestivum) in the background of the Arabidopsis tpc1-2 mutant. Cross-species tpc1 complementation and patch-clamping of vacuoles using Arabidopsis and rice tpc1 null mutants documented that both monocot TPC1 genes were capable of rescuing the SV channel deficit. Vacuoles from wild-type rice but not the tpc1 loss-of-function mutant harbor SV channels exhibiting the hallmark properties of dicot TPC1/SV channels. When expressed in human embryonic kidney (HEK293) cells OsTPC1 was targeted to Lysotracker-Red-positive organelles. The finding that the rice TPC1, just like those from the model plant Arabidopsis and even animal cells, is localized and active in lyso-vacuolar membranes associates this cation channel species with endomembrane function. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

  2. Analysis and optimization of energy resolution of neutron-TPC

    Institute of Scientific and Technical Information of China (English)

    黄孟; 李玉兰; 牛莉博; 李金; 李元景

    2015-01-01

    Neutron-TPC (nTPC) is a fast neutron spectrometer based on GEM-TPC (Gas Electron Multiplier-Time Pro-jection Chamber) and expected to be used in nuclear physics, nuclear reactor operation monitoring, and thermo-nuclear fusion plasma diagnostics. By measuring the recoiled proton energy and slopes of the proton tracks, the incident neutron energy can be deduced. It has higher n/γseparation ability and higher detection efficiency than conventional neutron spectrometers. In this paper, neutron energy resolution of nTPC is studied using the analytical method. It is found that the neutron energy resolution is determined by 1) the proton energy resolu-tion (σEp/Ep), and 2) standard deviation of slopes of the proton tracks caused by multiple Coulomb scattering (σk(scat ering)) and by the track fitting accuracy (σk(fit)). Suggestions are made for optimizing energy resolution of nTPC. Proper choices of the cut parameters of reconstructed proton scattering angles (θcut), the number of fitting track points (N ), and the working gas help to improve the neutron energy resolution.

  3. Free electron lifetime achievements in liquid Argon imaging TPC

    Energy Technology Data Exchange (ETDEWEB)

    Baibussinov, B; Ceolin, M Baldo; Centro, S; Cieslik, K; Farnese, C; Fava, A; Gibin, D; Guglielmi, A; Meng, G; Pietropaolo, F; Varanini, F; Ventura, S [INFN, Sezione di Padova via Marzolo 8, I-35131 Padova (Italy); Calligarich, E [INFN, Sezione di Pavia via Bassi 6, I-27100 Pavia (Italy); Rubbia, C, E-mail: Carlo.Rubbia@cern.c [Laboratori Nazionali del Gran Sasso dell' INFN I-67010 Assergi (Italy)

    2010-03-15

    A key feature for the success of the liquid Argon imaging TPC (LAr-TPC) technology is the industrial purification against electro-negative impurities, especially Oxygen and Nitrogen remnants, which have to be continuously kept at an exceptionally low level by filtering and recirculating liquid Argon. Improved purification techniques have been applied to a 120 liters LAr-TPC test facility in the INFN-LNL laboratory. Through-going muon tracks have been used to determine the free electron lifetime in liquid Argon against electro-negative impurities. The short path length here observed (30 cm) is compensated by the high accuracy in the observation of the specific ionization of cosmic ray muons at sea level as a function of the drift distance. A free electron lifetime of tau {approx} (21.4{sup +7.3}{sub -4.3}) ms, namely > 15.8 ms at 90% C.L. has been observed over several weeks under stable conditions, corresponding to a residual Oxygen equivalent of {approx} 15 ppt (part per trillion). At 500 V/cm, the free electron speed is 1.5 mm/mus. In a LAr-TPC a free electron lifetime in excess of 15 ms corresponds for instance to an attenuation of less than 20% after a drift path of 5 m, opening the way to the operation of the LAr-TPC with exceptionally long drift distances.

  4. Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors

    DEFF Research Database (Denmark)

    Khan, N.; Jeffers, M.; Kumar, S.;

    2008-01-01

    ) against a panel of rhHDAC (recombinant human HDAC) isoforms. Eight rhHDACs were expressed using a baculoviral system, and a Fluor de Lystrade mark (Biomol International) HDAC assay was optimized for each purified isoform. The potency and selectivity of ten HDACs on class I isoforms (rhHDAC1, rhHDAC2, rh...

  5. Resolution studies of a GEM-based TPC

    Energy Technology Data Exchange (ETDEWEB)

    Killenberg, M.

    2006-12-15

    Currently there are four different concept studies trying to optimise the detector for the requirements at the ILC. In three of these detector concepts a time projection chamber (TPC) is foreseen as the main tracking device. To achieve the intended spatial resolution of 100 {mu}m, micro pattern gas detectors (MPGD) are considered for gas amplification. The two different MPGDs discussed for the ILC TPC are Micro-Mesh Gaseous Detectors (Micromegas) and Gas Electron Multiplier foils (GEMs). The current thesis shows resolution studies with a TPC prototype equipped with a triple GEM readout structure. A hodoscope made up of silicon strip sensors gives a precision reference track, allowing an unbiased measurement of the spatial resolution. High statistics measurements have been conducted at the DESY test beam facility, which provides positrons with a tunable energy between 1 GeV and 6 GeV. Using the independent measurement of the hodoscope allows systematic studies of the homogeneity of the TPC's electric field. The fluctuations of the field in the chamber's central region were found to be {delta}E/E=8.10{sup -3}. Field distortions have been determined and corrected, reducing the remaining deviations to a level well below the spatial resolution of the TPC. One important task is to reduce the number of ions drifting back into the sensitive volume. Special GEM settings with minimised ion backdrift have been examined with respect to their influence on the spatial resolution and it was found that the spatial resolution is not degraded using these special settings. The TPC prototype has been operated in a 4 T magnetic field, provided by a superconducting solenoid located at DESY Hamburg. Again the spatial resolution measured with the ion backdrift optimised settings is compared to that achieved with nonoptimised settings. In both cases the measured resolution is approximately 130 {mu}m. (orig.)

  6. Magnetic field map for a large TPC prototype

    Energy Technology Data Exchange (ETDEWEB)

    Grefe, Christian

    2008-12-15

    A new e{sup +}e{sup -} linear collider with an energy of up to 1000 GeV is currently being planned: the International Linear Collider (ILC). It will allow high precision measurements of the Higgs boson and physics beyond the Standard Model. In the Large Detector Concept (LDC) -which is one of the proposed detector concepts for the ILC- a Time Projection Chamber (TPC) is intended as the main tracking device. Within the EUDET project a large TPC prototype is currently being built as an infrastructure to test different gas amplification and readout technologies. The prototype will be operated in a 1T superconducting solenoid magnet -the PCMAG- at the DESY testbeam area. In order to reach the best possible track reconstruction the magnetic field has to be known very precisely throughout the TPC volume. The magnetic field of PCMAG has been measured in July 2007. In this work the creation of a high precision field map from the measurements is presented. The magnet and modelling techniques for its magnetic field are described. A model of the magnet has been created as a best fit from the measurements and its limitations are investigated. The field map will be included in the reconstruction software for the TPC prototype. (orig.)

  7. Development of a TPC for an ILC Detector

    CERN Document Server

    Diener, R

    2012-01-01

    The ILD concept, one of two proposed detector concepts for the planned International Linear Collider (ILC), foresees a Time Projection Chamber (TPC) as the main tracking detector. The LCTPC (Linear Collider TPC) collaboration pursues R&D to develop such a TPC based on the best state-of-the-art technology. After tests with smaller prototypes, the current efforts focus on studies using a large prototype with a diameter of 770 mm and a length of 610 mm. This prototype can accommodate seven read-out modules of a size comparable to the ones that would be used in the final TPC. Several prototypes of modules using Micromegas or GEM structures as gas amplification were constructed and tested. Besides the traditional pad read-out, a pixel read-out based on the TimePix chip is studied in these tests with up to 8 TimePix chips on a board. The current status and future plans of the R&D are presented.

  8. Prototype of the ALICE Time Projection Chamber (TPC) Field-Cage

    CERN Multimedia

    2003-01-01

    The ALICE Time Projection Chamber (TPC) is the main particle tracking detector in ALICE. Charged particles crossing the gas of the TPC knock electons out of their atoms, which then drift in in the electric field. By measuring the arrival of electrons at the end of the chamber, the TPC will reconstruct the paths of the original charged particles.

  9. Adaptive molecular evolution of the two-pore channel 1 gene TPC1 in the karst-adapted genus Primulina (Gesneriaceae).

    Science.gov (United States)

    Tao, Junjie; Feng, Chao; Ai, Bin; Kang, Ming

    2016-12-01

    Limestone karst areas possess high floral diversity and endemism. The genus Primulina, which contributes to the unique calcicole flora, has high species richness and exhibit specific soil-based habitat associations that are mainly distributed on calcareous karst soils. The adaptive molecular evolutionary mechanism of the genus to karst calcium-rich environments is still not well understood. The Ca(2+)-permeable channel TPC1 was used in this study to test whether its gene is involved in the local adaptation of Primulina to karst high-calcium soil environments. Specific amplification and sequencing primers were designed and used to amplify the full-length coding sequences of TPC1 from cDNA of 76 Primulina species. The sequence alignment without recombination and the corresponding reconstructed phylogeny tree were used in molecular evolutionary analyses at the nucleic acid level and amino acid level, respectively. Finally, the identified sites under positive selection were labelled on the predicted secondary structure of TPC1. Seventy-six full-length coding sequences of Primulina TPC1 were obtained. The length of the sequences varied between 2220 and 2286 bp and the insertion/deletion was located at the 5' end of the sequences. No signal of substitution saturation was detected in the sequences, while significant recombination breakpoints were detected. The molecular evolutionary analyses showed that TPC1 was dominated by purifying selection and the selective pressures were not significantly different among species lineages. However, significant signals of positive selection were detected at both TPC1 codon level and amino acid level, and five sites under positive selective pressure were identified by at least three different methods. The Ca(2+)-permeable channel TPC1 may be involved in the local adaptation of Primulina to karst Ca(2+)-rich environments. Different species lineages suffered similar selective pressure associated with calcium in karst environments, and

  10. Handbook of purified gases

    CERN Document Server

    Schoen, Helmut

    2015-01-01

    Technical gases are used in almost every field of industry, science and medicine and also as a means of control by government authorities and institutions and are regarded as indispensable means of assistance. In this complete handbook of purified gases the physical foundations of purified gases and mixtures as well as their manufacturing, purification, analysis, storage, handling and transport are presented in a comprehensive way. This important reference work is accompanied with a large number of Data Sheets dedicated to the most important purified gases.  

  11. Analytical solutions for space charge fields in TPC drift volumes

    CERN Document Server

    Rossegger, S; Schnizer, B

    2011-01-01

    At high particle rates and high multiplicities, Time Projection Chambers can suffer from field distortions due to slow moving ions that accumulate within the drift volume. These variations modify the electron trajectory along the drift path, affecting the tracking performance of the detector. In order to calculate the track distortions due to an arbitrary space charge distribution in a TPC, novel representations of the Green's function for a TPC-like geometry were worked out. This analytical approach permits accurate predictions of track distortions due to an arbitrary space charge distribution (by solving the Langevin equation) as well as the possibility to benchmark common numerical methods to calculate such space charge fields. (C) 2011 Elsevier B.V. All rights reserved.

  12. The ICARUS 50 1 LAr TPC in the CERN $\

    CERN Document Server

    Arneodo, F; De Mitri, I; Mazza, D; Parlati, S; Petrera, S; Piano Mortari, G; Verdecchia, M; Benetti, P; Borio di Tigliole, A A; Calligarich, E; Cesana, E; Dolfini, R; Mauri, F; Montanari, C; Rappoldi, A; Raselli, G L; Terrani, M; Torre, P; Vignoli, C; Bettini, A; Carpanese, C; Centro, Sandro; Pepato, Adriano; Pietropaolo, F; Ventura, Sandro; Bonesini, M; Boschetti, B; Calvi, M; Curioni, A; Cavalli, D; Ferrari, A; Ferrari, P; Negri, P; Paganoni, M; Pullia, Antonio; Sala, P R; Ragazzi, S; Redaelli, N G; Tabarelli de Fatis, T; Terranova, F; Tonazzo, A; Casagrande, Federico; Cennini, P; Puccini, A; Rubbia, André; Rubbia, Carlo; Revol, Jean Pierre Charles; Cline, D; Matthey, C; Otwinowski, S; Park, J; Wang, H; Woo, J; Givoletti, J; Lamarina, A; Periale, R; Picchi, P; Mannocchi, G; Periale, L; Suzuki, S; Sergiampietri, F

    1999-01-01

    The 50 litre liquid Argon TPC is a detector built and successfully operated at CERN for R&D purposes within the ICARUS programme. In the year 1997 it has been exposed at the CERN neutrino beam for the entire SPS neutrino run period as proposed and approved at the SPSLC of January 1997. The detector, complemented with scintillators acting as veto, trigger counters and pre-shower counters, was installed in front of the NOMAD detector. The year 1997 was scheduled to be the last for the operation of the West Area Neutrino Facility. It was important to take this last opportunity for a parasitic exposure, which did not interfere with running experiments, of an already existing and operating liquid Argon TPC. As we had expected, the collected data brought important information for a better understanding of the performance of liquid Argon TPCs which should be useful for the entire ICARUS program. (9 refs).

  13. ARIADNE, a Photographic LAr TPC at the CERN Neutrino Platform

    CERN Document Server

    Mavrokoridis, K; Nessi, M; Roberts, A; Smith, N A; Touramanis, C; CERN. Geneva. SPS and PS Experiments Committee; SPSC

    2016-01-01

    This letter of intent describes a novel and innovative two-phase LAr TPC with photographic capabilities as an attractive alternative readout method to the currently accepted segmented THGEMs which will require many thousands of charge readout channels for kton-scale two-phase TPCs. These colossal LAr TPCs will be used for the future long-baseline-neutrino-oscillation experiments. Optical readout also presents many other clear advantages over current readout techniques such as ease of scalability, upgrade, installation and maintenance, and cost effectiveness. This technology has already been demonstrated at the Liverpool LAr facility with the photographic capturing of cosmic muon tracks and single gammas using a 40-litre prototype. We have now secured ERC funding to develop this further with the ARIADNE programme. ARIADNE will be a 1-ton two-phase LAr TPC utilizing THGEM and EMCCD camera readouts in order to photograph interactions, allowing for track reconstruction and particle identification. We are request...

  14. Upgrade of the ALICE-TPC read-out electronics

    Energy Technology Data Exchange (ETDEWEB)

    Junique, A; Mager, M; Musa, L; Rehman, A Ur, E-mail: Magnus.Mager@cern.ch [CERN, Geneva (Switzerland)

    2010-12-15

    The ALICE experiment at CERN LHC employs a large volume time projection chamber (TPC) as its main tracking device. Instigated by analyses indicating that the high level trigger is capable of sifting events with rare physics probes, it is endeavoured to read out the TPC an order of magnitude faster then was reckoned during the design of its read-out electronics. Based on an analysis of the read-out performance of the current system, an upgrade of the front-end read-out network is proposed. The performance of the foreseen architecture is simulated with raw data from real 7 TeV pp collisions. Events are superimposed in order to emulate the future ALICE running conditions: high multiplicity events generated either by PbPb collisions or by the superposition (pile-up) of a large number of pp collisions. The first prototype of the main building block has been produced and characterised, demonstrating the feasibility of the approach.

  15. Upgrade of the ALICE-TPC read-out electronics

    CERN Document Server

    Junique, A; Musa , L; Rehman , A U

    2010-01-01

    The ALICE experiment at CERN LHC employs a large volume time projection chamber (TPC) as its main tracking device. Instigated by analyses indicating that the high level trigger is capable of sifting events with rare physics probes, it is endeavoured to read out the TPC an order of magnitude faster then was reckoned during the design of its read-out electronics. Based on an analysis of the read-out performance of the current system, an upgrade of the front-end read-out network is proposed. The performance of the foreseen architecture is simulated with raw data from real 7 TeV pp collisions. Events are superimposed in order to emulate the future ALICE running conditions: high multiplicity events generated either by PbPb collisions or by the superposition (pile-up) of a large number of pp collisions. The first prototype of the main building block has been produced and characterised, demonstrating the feasibility of the approach

  16. Non-Uniform Electromagnetic Fields in the SAMURAI TPC

    Science.gov (United States)

    Estee, J.; Barney, J.; Chajecki, Z.; Chan, C. F.; Dunn, J. W.; Gilbert, J.; Lu, F.; Lynch, W. G.; Shane, R.; Tsang, M. B.; McIntosh, A. B.; Yennello, S. J.; Famiano, M.; Isobe, T.; Sakurai, H.; Taketani, A.; Murakami, T.; Samurai-Tpc Collaboration

    2011-10-01

    A Time Projection Chamber (TPC) is being developed for the SAMURAI dipole magnet at RIKEN. The main scientific objective for the TPC is to provide constraints on the nuclear symmetry at supra-saturation density. The poster presentation will discuss the design of the TPC field cage and the external electrodes that shape the high electric fields near the cathode. Garfield calculations of the electric field as well as TOSCA calculations of the magnetic field of the SAMURAI dipole will be shown and the impact of the non-uniformity of both fields on electron transport will be discussed. These non-uniformities can introduce components into the electron drift velocity in directions other than the expected vertical direction. This poster presentation will discuss the initial design of a laser calibration system, which will be used to calibrate away the influence of these non-uniformities in the electric and magnetic fields. This work is supported by the DOE under Grant DE-SC0004835.

  17. Pad Plane Design and Readout for SAMURAI TPC

    Science.gov (United States)

    Barney, J.; Chajecki, Z.; Chan, C. F.; Dunn, J. W.; Estee, J.; Gilbert, J.; Lu, F.; Lynch, W. G.; Shane, R.; Tsang, M. B.; McIntosh, A. B.; Yenello, S. J.; Famiano, M.; Isobe, T.; Sakurai, H.; Taketani, A.; Murakami, T.; Samurai-Tpc Collaboration

    2011-10-01

    The SAMURAI TPC is being built at Michigan State University to be used in the SAMURAI spectrometer at RIKEN in Japan, as part of the Symmetry Energy project, which focuses on obtaining constraints on the symmetry energy at supra-saturation densities. The presentation will discuss the development of the TPC as well as design for readout plane design for the TPC. These involve enabling the use of existing and future front end electronics (FEE), making the most of limited space, designing a circuit board for the pad plane, and techniques to glue the pad plane. The pad plane has been designed to work with either STAR or AGET electronics. The pad plane is made of a circuit board designed to minimize crosstalk and capacitance. The board must be built in smaller pieces and tiled, using alignment pins and precision gluing. Prototypes for the pad plane to FEE connection, pad plane gluing and STAR card mounting will be presented. Supported by the Department of Energy under Grant DE-SC0004835.

  18. Materials Testing and Performance Optimization for the SAMURAI-TPC

    Science.gov (United States)

    Long, K. D.; Lynch, W. G.; Barney, J.; Chajecki, Z.; Estee, J.; Shane, R.; Tangwanchareon, S.; Tsang, M. B.; Yurkon, J.

    2012-10-01

    The SAMURAI time-projection chamber (TPC) will be used to make measurements of pion spectra from heavy ion collisions at RIBF in Japan. Such research provides an opportunity to study supra-saturation density neutron-rich matter in the laboratory, and is critical to understanding the structure of neutron stars. It will provide a complete, 3D picture of the ionization deposited in a gas volume, from which particle types and momenta can be determined. The gas-containment volume is composed of surfaces of aluminum and plastic, as well as halogen-free printed circuit board. During multiplication of the ionized electrons at the anode wire plane of the TPC, UV photons are produced. These cause unwanted discharges when they interact with oxidized aluminum surfaces, which have low work functions. This problem can be addressed by application of a suitable conductive paint or epoxy. Paints were investigated to insure they did not contain any materials capable of inhibiting the performance of the detector gas. These investigations were cross-checked by tests carried out using an existing BRAHMS-TPC. Details on these tests and the materials chosen will be shown. The design and optimization of the gating grid, used to limit data collection to triggered events, will also be discussed.

  19. A continuous read-out TPC for the ALICE upgrade

    Science.gov (United States)

    Lippmann, C.

    2016-07-01

    The largest gaseous Time Projection Chamber (TPC) in the world, the ALICE TPC, will be upgraded based on Micro Pattern Gas Detector technology during the second long shutdown of the CERN Large Hadron Collider in 2018/19. The upgraded detector will operate continuously without the use of a triggered gating grid. It will thus be able to read all minimum bias Pb-Pb events that the LHC will deliver at the anticipated peak interaction rate of 50 kHz for the high luminosity heavy-ion era. New read-out electronics will send the continuous data stream to a new online farm at rates up to 1 TByte/s. A fractional ion feedback of below 1% is required to keep distortions due to space charge in the TPC drift volume at a tolerable level. The new read-out chambers will consist of quadruple stacks of Gas Electron Multipliers (GEM), combining GEM foils with a different hole pitch. Other key requirements such as energy resolution and operational stability have to be met as well. A careful optimisation of the performance in terms of all these parameters was achieved during an extensive R&D program. A working point well within the design specifications was identified with an ion backflow of 0.63%, a local energy resolution of 11.3% (sigma) and a discharge probability comparable to that of standard triple GEM detectors.

  20. Slow Control System for the NIFFTE Collaboration TPC

    Science.gov (United States)

    Ringle, Erik; Niffte Collaboration Collaboration

    2011-10-01

    As world energy concerns continue to dominate public policy in the 21st century, the need for cleaner and more efficient nuclear power is necessary. In order to effectively design and implement plans for generation IV nuclear reactors, more accurate fission cross-section measurements are necessary. The Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) collaboration, in an effort to meet this need, has constructed a Time Projection Chamber (TPC) which aims to reduce the uncertainty of the fission cross-section to less than 1%. Using the Maximum Integration Data Acquisition System (MIDAS) framework, slow control measurements are integrated into a single interface to facilitate off-site monitoring. The Hart Scientific 1560 Black Stack will be used with two 2564 Thermistor Scanner Modules to monitor internal temperature of the TPC. A Prologix GPIB to Ethernet controller will be used to interface the hardware with MIDAS. This presentation will detail the design and implementation of the slow control system for the TPC. This work was supported by the U.S. Department of Energy Division of Energy Research.

  1. Free electron lifetime achievements in Liquid Argon Imaging TPC

    CERN Document Server

    Baibussinov, B; Calligarich, E; Centro, S; Cieslik, K; Farnese, C; Fava, A; Gibin, D; Guglielmi, A; Meng, G; Pietropaolo, F; Rubbia, C; Varanini, F; Ventura, S

    2010-01-01

    A key feature for the success of the Liquid Argon TPC technology is the industrial purification against electro-negative impurities, especially Oxygen and Nitrogen remnants, which have to be initially and continuously kept at an exceptional purity. New purification techniques have been applied to a 120 litres LAr-TPC test facility in the INFN-LNL laboratory. Through-going muon tracks have been used to monitor the LAr purity. The short path length used (30 cm) is compensated by the high accuracy in the observation of the specific ionization of cosmic rays muons at sea level. A free electron lifetime of (21.4+7.3-4.3) ms, namely > 15.8 ms at 90 % C.L. has been observed under stable conditions over several weeks, corresponding to about 15 ppt (part per trillion) of Oxygen equivalent. At 500 V/cm, where the electron speed is approximately of 1.5 mm/us, the free electron lifetime >15 ms corresponds to an attenuation <15 % for a drift path of 5 m, opening the way to reliable operation of LAr TPC for exceptionall...

  2. Future Upgrade and Physics Perspectives of the ALICE TPC

    CERN Document Server

    INSPIRE-00033137

    2014-01-01

    The ALICE experiment at the Large Hadron Collider (LHC) proposes major detector upgrades to fully exploit the increase of the luminosity of the LHC in RUN~3 and to extend the physics reach for rare probes at low transverse momentum. The Time Projection Chamber (TPC) is one of the main tracking and PID devices in the central barrel of ALICE. The maximum trigger rate of the TPC is currently limited to about 3.5 kHz by the operation of a gating grid system. In order to make full use of the luminosity in RUN 3, the TPC is foreseen to be operated in an ungated mode with continuous readout. The existing MWPC readout will be replaced by a Micro-Pattern Gaseous Detector (MPGD) based readout, which provides intrinsic ion capture capability without gating. Extensive detector R\\&D employing Gas Electron Multiplier (GEM) and Micro-Mesh Gaseous detector (Micromegas) technologies, and simulation studies to advance the techniques for the corrections of space-charge distortions have been performed since 2012. In this pap...

  3. The Readout Control Unit of the ALICE TPC

    CERN Document Server

    Lien, J A; Musa, L

    2004-01-01

    The ALICE Time Projection Chamber (TPC) is the main tracking detector of the central barrel of the ALICE (A Large Ion Collider) Experiment at the Large Hadron Collider (LHC), being constructed at CERN, Geneva. It is a 88 m$^{3}$ cylinder filled with gas and divided into two drift regions by the central electrode located at its axial center. The readout chambers of the TPC are multi-wire proportional chambers with cathode pad readout. About 570 000 pads are read-out by an electronics chain of amplification, digitalization and pre-processing. One of the challenges in designing the TPC for ALICE is the design of Front End Electronics (FEE) to cope with the data rates and the channel occupancy. The Readout Control Unit (RCU), which is presented in this work, is designed to control and monitor the Front End Electronics, and to collect and ship data to the High Level Trigger and the Data Acquisition System, via the Detector Data Link (DDL - optical fibre). The RCU must be capable of reading out up to 200 Mbytes/s f...

  4. Purified water quality study

    Energy Technology Data Exchange (ETDEWEB)

    Spinka, H.; Jackowski, P.

    2000-04-03

    Argonne National Laboratory (HEP) is examining the use of purified water for the detection medium in cosmic ray sensors. These sensors are to be deployed in a remote location in Argentina. The purpose of this study is to provide information and preliminary analysis of available water treatment options and associated costs. This information, along with the technical requirements of the sensors, will allow the project team to determine the required water quality to meet the overall project goals.

  5. SAMURAI-TPC: Field Cage Design and Prototyping

    Science.gov (United States)

    Lu, F.; Barney, J.; Chajecki, Z.; Chan, C. F.; Dunn, J. W.; Estee, J.; Gilbert, J.; Lynch, W. G.; Shane, R.; Tsang, M. B.; McIntosh, A. B.; Yenello, S. J.; Famiano, M.; Isobe, T.; Sakurai, H.; Taketani, A.; Murakami, T.; Samurai-Tpc Collaboration

    2011-10-01

    The SAMURAI-TPC is a time-projection chamber to be used in conjunction with the SAMURAI spectrometer being built at the Radioactive Isotope Beam Facility at RIKEN, Japan. It will be used to measure charged pions, protons and light ions. The pi+/pi- ratios from heavy-ion collisions should provide constraints on the asymmetry term in the nuclear equation of state at densities about twice saturation density. In this talk, the design and operation of the field cage, an essential part of the detector, will be discussed, along with the results of prototype testing. This work is supported by the Department of Energy under Grant DE-SC0004835.

  6. A 4. pi. tracking TPC magnetic spectrometer for RHIC

    Energy Technology Data Exchange (ETDEWEB)

    Danby, G.; Eiseman, S.E.; Etkin, A.; Foley, K.J.; Hackenburg, R.W.; Longacre, R.S.; Love, W.A.; Morris, T.W.; Platner, E.D.; Saulys, A.C.; Van Dijk, J.H. (Brookhaven National Lab., Upton, NY (USA)); Lindenbaum, S.J. (Brookhaven National Lab., Upton, NY (USA) City Coll., New York, NY (USA)); Chan, C.S.; Kramer, M.A.; Zhao, K. (City Coll., New York, NY (USA)); Biswas, N.; Kenney, P.; Piekarz, J. (Notre Dame Univ

    1990-01-01

    The primary physics objective of the 4{pi} TPC magnetic spectrometer proposal is to search for the Quark-Gluon Plasma. In previous workshops we have discussed what the possible hadronic signatures of such a state of matter would be. Succinctly, the QGP is a direct prediction of non-perturbative QCD. Therefore the question of the existence of this new state of matter bears directly on the validity of non-perturbative QCD. However, since non-perturbative QCD has never been established, it is apparent that what may await us is a host of new phenomena that will go beyond the standard model.

  7. Electron transmission efficiency of gating-GEM foil for TPC

    Institute of Scientific and Technical Information of China (English)

    XIE Wen-Qing; HUANG Meng; LI Ting; TIAN Yang; LI Yu-Lan; LI Yuan-Jing

    2012-01-01

    In a TPC,ion feedback from the readout detector can cause a space-charge effect and distort the electrical field in the drift region.Gating is one of the effective methods to solve this problem,which can block ions at the expense of losing a certain amount of primary electrons.Compared with the traditional design with a wire structure,gating based on GEM foil is more attractive because of its simplicity.In this paper,the factors influencing the electron transmission efficiency are studied with simulations and experiments.After optimizing all these parameters,an electron transmission efficiency over 80% is obtained.

  8. Amyloid-beta Isoform Metabolism Quantitation by Stable Isotope Labeled Kinetics

    OpenAIRE

    Mawuenyega, Kwasi G.; Kasten, Tom; Sigurdson, Wendy; Bateman, Randall J.

    2013-01-01

    Abundant evidence suggests a central role for the amyloid-β (Aβ) peptide in Alzheimer’s disease (AD) pathogenesis. Production and clearance of different Aβ isoforms have been established as targets of proposed disease-modifying therapeutic treatments of AD. However, previous studies used multiple sequential purification steps to isolate the isoforms individually and quantitate them based on a common mid-domain peptide. We created a method to simultaneously purifyisoforms and quantitate th...

  9. Cold Electronics Development for the LBNE LAr TPC

    Science.gov (United States)

    Thorn, C.; De Geronimo, Gianluigi; D'Andragora, Alessio; Li, Shaorui; Nambiar, Neena; Rescia, Sergio; Vernon, Emerson; Chen, Hucheng; Lanni, Francesco; Makowiecki, Don; Radeka, Veljko; Yu, Bo

    The LBNE Project is developing a design for multiple 20 kiloton liquid argon (LAr) time projection chambers to be used as the far detector for the Long Baseline Neutrino Experiment. An essential component of this design is a complete electronic readout system designed to operate in LAr (at 90K). This system is being implemented as a CMOS ASIC, in 180 nm commercial technology, that will provide low-noise readout of the signals induced on the TPC wires, digitization of those signals at 2 MS/s, zero-suppression, buffering and output multiplexing to a small number of cryostat feed-throughs. A resolution better than 1000 rms electrons at 200 pF input capacitance for an input range of 300 fC is required, along with low power (analog-only frontend has been successfully completed and fully evaluated, and will be used in the MicroBooNE LAr TPC. A prototype of the digital section has been fabricated and is being evaluated. The results demonstrate that CMOS transistors have lower noise and much improved dc characteristics at LAr temperature. We will describe the progress to date and plans for the remaining development.

  10. Software and Parameters for Detailed TPC Studies in the CLIC CDR

    CERN Document Server

    Killenberg, M.

    2011-01-01

    For the TPC occupancy and time stamping studies in the CLIC CDR the MarlinTPC software package has been used in combination with Mokka for the full detector simulation. This document describes the working principle of the Marlin processors used for digitisation and reconstruction, and lists the parameters for reference.

  11. Heavy ion reaction measurements with the EOS TPC (looking for central collisions with missing energy)

    Energy Technology Data Exchange (ETDEWEB)

    Wieman, H.H.; EOS Collaboration

    1994-05-01

    The EOS TPC was constructed for complete event measurement of heavy ion collisions at the Bevalac. We report here on the TPC design and some preliminary measurements of conserved event quantities such as total invariant mass, total momentum, total A and Z.

  12. Natural Air Purifier

    Science.gov (United States)

    1993-01-01

    NASA environmental research has led to a plant-based air filtering system. Dr. B.C. Wolverton, a former NASA engineer who developed a biological filtering system for space life support, served as a consultant to Terra Firma Environmental. The company is marketing the BioFilter, a natural air purifier that combines activated carbon and other filter media with living plants and microorganisms. The filter material traps and holds indoor pollutants; plant roots and microorganisms then convert the pollutants into food for the plant. Most non-flowering house plants will work. After pollutants have been removed, the cleansed air is returned to the room through slits in the planter. Terra Firma is currently developing a filter that will also disinfect the air.

  13. The liquid argon TPC for the ICARUS experiment

    CERN Document Server

    Arneodo, F

    1997-01-01

    The ICARUS project aims at the realisation of a large liquid argon TPC to be run at the Underground Laboratories of Gran Sasso in Italy. An intense R&D; activity has put on firm grounds this new detector technology and experimentally confirmed its feasibility on a few ton scale. Based on these solid achievements, the collaboration is now confident of being able to build and safely operate a multi-kton detector. The reseach program of the experiment involves the systematic study of a wide spectrum of physical phenomena covering many orders of magnitude in the energy deposited in the detector: from the few MeV of solar neutrino interactions, to the about one GeV of the proton decay and atmospheric neutrinos, up to the higher energies of neutrinos from accelerators.

  14. NEXT, a HPGXe TPC for neutrinoless double beta decay searches

    CERN Document Server

    Granena, F; Nova, F; Rico, J; Sánchez, F; Nygren, D R; Barata, J A S; Borges, F I G M; Conde, C A N; Dias, T H V T; Fernandes, L M P; Freitas, E D C; Lopes, J A M; Monteiro, C M B; Santos, J M F dos; Santos, F P; Tavora, L M N; Veloso, J F C A; Calvo, E; Gil-Botella, I; Novella, P; Palomares, C; Verdugo, A; Giomataris, Yu; Ferrer-Ribas, E; Hernando-Morata, J A; Martínez, D; Cid, X; Ball, M; Carcel, S; Cervera-Villanueva, Anselmo; Díaz, J; Gil, A; Gómez-Cadenas, J J; Martín-Albo, J; Monrabal, F; Munoz-Vidal, J; Serra, L; Sorel, M; Yahlali, N; Bosch, R Esteve; Lerche, C W; Martinez, J D; Mora, F J; Sebastiá, A; Tarazona, A; Toledo, J F; Lazaro, M; Perez, J L; Ripoll, L; Carmona, J M; Cebrián, S; Dafni, T; Galan, J; Gomez, H; Iguaz, F J; Irastorza, I G; Luzón, G; Morales, J; Rodríguez, A; Ruz, J; Tomas, A; Villar, J A

    2009-01-01

    We propose a novel detection concept for neutrinoless double-beta decay searches. This concept is based on a Time Projection Chamber (TPC) filled with high-pressure gaseous xenon, and with separated-function capabilities for calorimetry and tracking. Thanks to its excellent energy resolution, together with its powerful background rejection provided by the distinct double-beta decay topological signature, the design discussed in this Letter Of Intent promises to be competitive and possibly out-perform existing proposals for next-generation neutrinoless double-beta decay experiments. We discuss the detection principles, design specifications, physics potential and R&D plans to construct a detector with 100 kg fiducial mass in the double-beta decay emitting isotope Xe(136), to be installed in the Canfranc Underground Laboratory.

  15. Topological signature in the NEXT high pressure xenon TPC

    CERN Document Server

    ,

    2016-01-01

    The NEXT experiment aims to observe the neutrinoless double beta decay of Xe-136 in a high-pressure xenon gas TPC using electroluminescence to amplify the signal from ionization. One of the main advantages of this technology is the possibility to use the topology of events with energies close to Qbb as an extra tool to reject background. In these proceedings we show with data from prototypes that an extra background rejection factor of 24.3 +- 1.4 (stat.)% can be achieved, while maintaining an efficiency of 66.7 +- 1.% for signal events. The performance expected in NEW, the next stage of the experiment, is to improve to 12.9% +- 0.6% background acceptance for 66.9% +- 0.6% signal efficiency.

  16. NEXT: R and D towards a xenon high pressure TPC

    Energy Technology Data Exchange (ETDEWEB)

    Lux, Thorsten [Universitat Autonoma de Barcelona, Barcelona (Spain); Sanchez, Federico [IFAE, Barcelona (Spain); Gomez-Cadenas, J.J.; Martin-Albo, Justo; Ball, Markus; Novella, Pau; Monrabal, Francesc; Cervera, Anselmo [IFIC, Valencia (Spain); Garcia Irastorza, Igor [Universidad de Zaragoza, Zaragoza (Spain)

    2008-07-01

    An open question within the Standard Model is the nature of the neutrino. Is it a Majorana or a Dirac particle? The only way to answer this, is the search for neutrino-less double beta decays. Various experimental approaches are investigated for this reason e.g. diodes, bolometers, liquid Xenon. The key points for all of them is the high requirements on the energy resolution to distinguish between the decay with two neutrinos and the neutrino-less decay and the external background suppression. Recently some Spanish groups started a R and D program to investigate the possibility to use a pressurized Xenon TPC with MPGD readout (MM, LEM (GEM)). In the presentation the choice of gaseous Xe is motivated and an overview about the R and D plans is given.

  17. A TPC for sPHENIX at RHIC

    Science.gov (United States)

    Ramasubramanian, Niveditha; Dehmelt, Klaus; Sphenix Collaboration

    2016-09-01

    The sPHENIX detector is being proposed at the Relativistic Heavy Ion Collider to measure jets and upsilons for advancing our understanding of the quark gluon plasma formed in heavy ion collisions. It is also expected to form the basis of a day-1 detector for a future U.S. Electron Ion Collider. sPHENIX is based on a superconducting solenoidal magnet formerly used by the BaBar experiment, and of charged particle tracking, electromagnetic as well as hadronic calorimetry. It covers a large acceptance, 2 π in azimuth and pseudorapidities of | η | < 1, and allows to acquire data at a rate of up to 15 kHz. Furthermore, a Gas Electron Multiplier based Time Projection Chamber has been proposed to improve tracking resolution in a high multiplicity environment. In this talk we will present the current design and status of ongoing R&D and simulation studies for tracking with a TPC.

  18. Readout of TPC Tracking Chambers with GEMs and Pixel Chip

    Energy Technology Data Exchange (ETDEWEB)

    Kadyk, John; Kim, T.; Freytsis, M.; Button-Shafer, J.; Kadyk, J.; Vahsen, S.E.; Wenzel, W.A.

    2007-12-21

    Two layers of GEMs and the ATLAS Pixel Chip, FEI3, have been combined and tested as a prototype for Time Projection Chamber (TPC) readout at the International Linear Collider (ILC). The double-layer GEM system amplifies charge with gain sufficient to detect all track ionization. The suitability of three gas mixtures for this application was investigated, and gain measurements are presented. A large sample of cosmic ray tracks was reconstructed in 3D by using the simultaneous timing and 2D spatial information from the pixel chip. The chip provides pixel charge measurement as well as timing. These results demonstrate that a double GEM and pixel combination, with a suitably modified pixel ASIC, could meet the stringent readout requirements of the ILC.

  19. Absolute quantitation of protein posttranslational modification isoform.

    Science.gov (United States)

    Yang, Zhu; Li, Ning

    2015-01-01

    -site-independent peptides in the total cellular protein and their peptide yields. The PTM occupancy determination is achieved by measuring the absolute amounts of both PTM and non-PTM peptides from the highly purified protein sample expressed in transgenic organisms or directly isolated from an organism using affinity purification. The absolute amount of each PTM isoform in the total cellular protein extract is finally calculated from these two variables. Following this approach, the ion intensities given by mass spectrometers are used to calculated the peptide amounts, from which the amounts of protein isoforms are then deduced. In this chapter, we describe the principles underlying the experimental design and procedures used in AQUIP method. This quantitation method basically employs stable isotope-labeled peptide standards and affinity purification from a tagged recombinant protein of interest. Other quantitation strategies and purification techniques related to this method are also discussed.

  20. SAMURAI-TPC: A Time Projection Chamber for Constraining the Asymmetry Energy at High Density

    Science.gov (United States)

    McIntosh, A. B.; Maass, N.; Yennello, S. J.; Barney, J.; Chajecki, Z.; Chan, C. F.; Dunn, J. W.; Estee, J.; Gilbert, J.; Lu, F.; Lynch, W. G.; Shane, R.; Tsang, M. B.; Famiano, M.; Isobe, T.; Sakurai, H.; Taketani, A.; Murakami, T.; Samurai-Tpc Collaboration

    2011-10-01

    The SAMURAI-TPC is a time projection chamber designed to measure pions and light charged particles. By measuring pion yield ratios and particle flow in heavy ion collisions around E = 200A MeV, we expect to constrain the behavior of the nuclear asymmetry energy around twice saturation density. In this talk, the design and construction of the TPC components will be discussed. Upon completion, the SAMURAI-TPC will be installed in the SAMURAI spectrometer at the Radioactive Isotope Beam Facility at RIKEN, Japan. This work is supported by the Department of Energy (DE-SC0004835).

  1. Device for purifying drilling mud

    Energy Technology Data Exchange (ETDEWEB)

    Surkov, V.T.; Dorosh, M.M.; Khariv, I.Yu.; Makedonov, N.I.

    1982-01-01

    A device is proposed for purifying drilling mud which includes a dynamic filter made in the form of a spiral-shaped tube with input and output sleeves, and a container for purified solution with outlet sleeve. It is distinguished by the fact that in order to simplify the design, the spiral-shaped tube is perforated from the inside and is installed in the container for the purified solution.

  2. Sealed operation, and circulation and purification of gas in the HARPO TPC

    CERN Document Server

    Frotin, M; Attié, D; Bernard, D; Dauvois, V; Delbart, A; Durand, D; Geerebaert, Y; Legand, S; Magnier, P; Poilleux, P; Semeniouk, I

    2015-01-01

    HARPO is a time projection chamber (TPC) demonstrator of a gamma-ray telescope and polarimeter in the MeV-GeV range, for a future space mission. We present the evolution of the TPC performance over a five month sealed-mode operation, by the analysis of cosmic-ray data, followed by the fast and complete recovery of the initial gas properties using a lightweight gas circulation and purification system.

  3. Optically read out GEM-based TPC operation and preliminary scintillation studies

    CERN Document Server

    Galgoczi, Gabor

    2016-01-01

    The main goal of this project was to realise the reconstruction of tracks in an optically read out GEM (Gas Electron Multiplier) based Time Projection Chamber (TPC). Secondary goal was to initialise a series of systematic studies on the scintillation of particles in Ar/CF4 (80-20%) mixture. Track reconstruction is needed for primary scintillation studies as only tracks fully contained can be considered. A vetoing and trigerring logic was built for the TPC from NIM modules.

  4. Behavior of TPC`s in a high particle flux environment

    Energy Technology Data Exchange (ETDEWEB)

    Etkin, A.; Eisemann, S.E.; Foley, K.J.; Hackenburg, R.W.; Longacre, R.S.; Love, W.A.; Morris, T.W.; Platner, E.D.; Saulys, A.C. [Brookhaven National Lab., Upton, NY (United States); Lindenbaum, S.J. [Brookhaven National Lab., Upton, NY (United States)]|[City Coll., New York, NY (United States); Chan, C.S.; Kramer, M.A.; Zhao, K.H.; Zhu, Y. [City Coll., New York, NY (United States); Hallman, T.J.; Madansky, L. [Johns Hopkins Univ., Baltimore, MD (United States); Ahmad, S.; Bonner, B.E.; Buchanan, J.A.; Chiou, C.N.; Clement, J.M.; Mutchler, G.S.; Roberts, J.B. [Rice Univ., Houston, TX (United States). Bonner Nuclear Labs.

    1991-12-13

    TPC`s (Time Projection Chamber) used in E-810 at the AGS (Alternating Gradient Synchrotron) were exposed to fluxes equivalent to more than 10{sup 7} minimum ionizing particles per second to find if such high fluxes cause gain changes or distortions of the electric field. Initial results of these and other tests are presented and the consequences for the RHIC (Relativistic Heavy Ion Collider) TPC-based experiments are discussed.

  5. Behavior of TPC`s in a high particle flux environment

    Energy Technology Data Exchange (ETDEWEB)

    Etkin, A.; Eiseman, S.E.; Foley, K.J.; Hackenburg, R.W.; Longacre, R.S.; Love, W.A.; Morris, T.W.; Platner, E.D.; Saulys, A.C. [Brookhaven National Lab., Upton, NY (United States); Lindenbaum, S.J. [Brookhaven National Lab., Upton, NY (United States)]|[City Coll., New York, NY (United States); Chan, C.S.; Kramer, M.A.; Zhao, K.H.; Zhu, Y. [City Coll., New York, NY (United States); Hallman, T.J.; Madansky, L. [Johns Hopkins Univ., Baltimore, MD (United States); Ahmad, S.; Bonner, B.E.; Buchanan, J.A.; Chiou, C.N.; Clement, J.M.; Mutchler, G.S.; Roberts, J.B. [Rice Univ., Houston, TX (United States)

    1992-07-08

    TPC`s (Time Projection Chamber) used in E-810 at the AGS (Alternating Gradient Synchrotron) were exposed to fluxes equivalent to more than 10{sup 7} minimum ionizing particles per second to find if such high fluxes cause gain changes or distortions of the electric field. Initial results of these and other tests are presented and the consequences for the RHIC (Relativistic Heavy Ion collider) TPC-based experiments are discussed.

  6. Behavior of TPC`s in a high particle flux environment

    Energy Technology Data Exchange (ETDEWEB)

    Etkin, A.; Eiseman, S.E.; Foley, K.J.; Hackenburg, R.W.; Longacre, R.S.; Love, W.A.; Morris, T.W.; Platner, E.D.; Saulys, A.C.; Lindenbaum, S.J. [Brookhaven National Lab., Upton, NY (United States); Chan, C.S.; Kramer, M.A.; Zhao, K.H.; Zhu, Y. [City College of New York, New York (United States); Hallman, T.J.; Madansky, L. [Johns Hopkins Univ., Baltimore, MD (United States); Ahmad, S.; Bonner, B.E.; Buchanan, J.A.; Chiou, C.N.; Clement, J.M.; Mutchler, G.S.; Roberts, J.B. [Bonner Nuclear Lab., Houston, TX (United States)

    1991-12-31

    TPC`s (Time Projection Chamber) used in E-810 at the TAGS (Alternating Gradient Synchrotron) were exposed to fluxes equivalent to more than 10 minimum ionizing particles per second to find if such high fluxes cause gain changes or distortions of the electric field. Initial results of these and other tests are presented and the consequences for the RHIC (Relativistic Heavy Ion Collider) TPC-based experiments are discussed.

  7. Behavior of TPC`s in a high particle flux environment

    Energy Technology Data Exchange (ETDEWEB)

    Etkin, A.; Eiseman, S.E.; Foley, K.J.; Hackenburg, R.W.; Longacre, R.S.; Love, W.A.; Morris, T.W.; Platner, E.D.; Saulys, A.C. [Brookhaven National Lab., Upton, NY (United States); Lindenbaum, S.J. [Brookhaven National Lab., Upton, NY (United States)]|[City Coll., New York, NY (United States); Chan, C.S.; Kramer, M.A.; Zhao, K.H.; Zhu, Y. [City Coll., New York, NY (United States); Hallman, T.J.; Madansky, L. [Johns Hopkins Univ., Baltimore, MD (United States); Ahmad, S.; Bonner, B.E.; Buchanan, J.A.; Chiou, C.N.; Clement, J.M.; Mutchler, G.S.; Roberts, J.B. [Rice Univ., Houston, TX (United States). Bonner Nuclear Labs.

    1991-12-31

    TPC`s (Time Projection Chamber) used in E-810 at the AGS (Alternating Gradient Synchrotron) were exposed to fluxes equivalent to more than 10{sup 7} minimum ionizing particles per second to find if such high fluxes cause gain changes or distortions of the electric field. Initial results of these and other tests are presented and the consequences for the RHIC (Relativistic Heavy Ion Collider) TPC-based experiments are discussed.

  8. ALICE TPC upgrade for High-Rate operations

    CERN Document Server

    ,

    2015-01-01

    A new type of Time Projection Chamber (TPC) has been proposed for the upgrade of the ALICE (A Large Ion Collider Experiment at CERN) so as to cater to the high luminosity environment expected at the Large Hadron Collider (LHC) facility in future. This device will rely on the intrinsic ion back flow (IBF) suppression of Micro-Pattern Gas Detectors (MPGD) based technology in particular the Gas Electron Multiplier (GEM). GEM is to minimise the space charge effect in the main drift volume and thus will not require the standard gating grid and the resulting intrinsic dead time. It will thus be possible to read all minimum bias Pb--Pb events that the Large Hadron Collider (LHC) will deliver at the anticipated peak interaction rate of 50 kHz for the high luminosity heavy-ion era in Run 3. New read-out electronics will send the continuous data stream to a new online farm at rates up to 1~TByte/s. The new read-out chambers will consist of stacks of 4 GEM foils combining different hole pitches. In addition to a low ion...

  9. AN OVERVIEW ON BLOOD PURIFIER

    Directory of Open Access Journals (Sweden)

    Sabia Chauhan

    2013-09-01

    Full Text Available Blood is a connective tissue which protects us from different problems. Without blood body cannot functions at all and blood doesn’t purify itself. When blood does not purifies itself that times kidney, liver and lymphatic system work together that they help’s to purifiers the blood. Causes which are included in blood impurities are modern life style, junk food, alcohol etc. If the blood becomes impure it causes different problems e.g. acne, rashes, allergic etc. There is not any proper synthetic medication for blood impurities. Only herbal formulations are used for the blood purifier. In this review article we discussed about the market formulations and the different plants which are used in them with their different activities which are helpful in purifies the blood and also protects from other problems.

  10. Two pore channel 2 (TPC2) inhibits autophagosomal-lysosomal fusion by alkalinizing lysosomal pH.

    Science.gov (United States)

    Lu, Yingying; Hao, Bai-Xia; Graeff, Richard; Wong, Connie W M; Wu, Wu-Tian; Yue, Jianbo

    2013-08-16

    Autophagy is an evolutionarily conserved lysosomal degradation pathway, yet the underlying mechanisms remain poorly understood. Nicotinic acid adenine dinucleotide phosphate (NAADP), one of the most potent Ca(2+) mobilizing messengers, elicits Ca(2+) release from lysosomes via the two pore channel 2 (TPC2) in many cell types. Here we found that overexpression of TPC2 in HeLa or mouse embryonic stem cells inhibited autophagosomal-lysosomal fusion, thereby resulting in the accumulation of autophagosomes. Treatment of TPC2 expressing cells with a cell permeant-NAADP agonist, NAADP-AM, further induced autophagosome accumulation. On the other hand, TPC2 knockdown or treatment of cells with Ned-19, a NAADP antagonist, markedly decreased the accumulation of autophagosomes. TPC2-induced accumulation of autophagosomes was also markedly blocked by ATG5 knockdown. Interestingly, inhibiting mTOR activity failed to increase TPC2-induced autophagosome accumulation. Instead, we found that overexpression of TPC2 alkalinized lysosomal pH, and lysosomal re-acidification abolished TPC2-induced autophagosome accumulation. In addition, TPC2 overexpression had no effect on general endosomal-lysosomal degradation but prevented the recruitment of Rab-7 to autophagosomes. Taken together, our data demonstrate that TPC2/NAADP/Ca(2+) signaling alkalinizes lysosomal pH to specifically inhibit the later stage of basal autophagy progression.

  11. Building a Class-1 Glove Box for Use with the NIFFTE TPC

    Science.gov (United States)

    Lynn, William

    2012-10-01

    The Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) uses a Time Projection Chamber (TPC) to measure the neutron-induced fission cross sections of actinides with unprecedented accuracy which will aid in the development of the next generation nuclear reactors. Charged particles, including fission fragments, create a trail of electrons within a fill gas through ionization, which then drift in an electric field towards the read-out electronics. Using a MicroMegas mesh, the signal is amplified and then detected by the TPC pad plane. Due to the delicate nature of the MicroMegas mesh, precaution must be taken to prevent damage to the mesh from airborne contaminants which can cause the mesh to short. To avoid radiological contamination, a glove box was chosen for the task of handling and installing actinide targets into the TPC. To protect the TPC electronics, a decision was made to modify the existing glove box to create a Class-1 cleanroom environment. Variables such as glove type, filter, and cleaning agent were tested independently to determine maximum cleanliness, and a procedure for creating an acceptable Class-1 environment inside the glove box for the TPC was developed.

  12. Noise Characterization and Filtering in the MicroBooNE Liquid Argon TPC

    Energy Technology Data Exchange (ETDEWEB)

    Acciarri, R.; et al.

    2017-05-20

    The low-noise operation of readout electronics in a liquid argon time projection chamber (LArTPC) is critical to properly extract the distribution of ionization charge deposited on the wire planes of the TPC, especially for the induction planes. This paper describes the characteristics and mitigation of the observed noise in the MicroBooNE detector. The MicroBooNE's single-phase LArTPC comprises two induction planes and one collection sense wire plane with a total of 8256 wires. Current induced on each TPC wire is amplified and shaped by custom low-power, low-noise ASICs immersed in the liquid argon. The digitization of the signal waveform occurs outside the cryostat. Using data from the first year of MicroBooNE operations, several excess noise sources in the TPC were identified and mitigated. The residual equivalent noise charge (ENC) after noise filtering varies with wire length and is found to be below 400 electrons for the longest wires (4.7 m). The response is consistent with the cold electronics design expectations and is found to be stable with time and uniform over the functioning channels. This noise level is significantly lower than previous experiments utilizing warm front-end electronics.

  13. Enhanced trigger for the NIFFTE fissionTPC in presence of high-rate alpha backgrounds

    Science.gov (United States)

    Bundgaard, Jeremy; Niffte Collaboration

    2015-10-01

    Nuclear physics and nuclear energy communities call for new, high precision measurements to improve existing fission models and design next generation reactors. The Neutron Induced Fission Fragment Tracking experiment (NIFFTE) has developed the fission Time Projection Chamber (fissionTPC) to measure neutron induced fission with unrivaled precision. The fissionTPC is annually deployed to the Weapons Neutron Research facility at Los Alamos Neutron Science Center where it operates with a neutron beam passing axially through the drift volume, irradiating heavy actinide targets to induce fission. The fissionTPC was developed at the Lawrence Livermore National Laboratory's TPC lab, where it measures spontaneous fission from radioactive sources to characterize detector response, improve performance, and evolve the design. To measure 244Cm, we've developed a fission trigger to reduce the data rate from alpha tracks while maintaining a high fission detection efficiency. In beam, alphas from 239Pu are a large background when detecting fission fragments; implementing the fission trigger will greatly reduce this background. The implementation of the cathode fission trigger in the fissionTPC will be presented along with a detailed study of its efficiency.

  14. A study with a small prototype TPC for the international linear collider experiment

    Energy Technology Data Exchange (ETDEWEB)

    Ackermann, K. [Max-Planck-Institute for Physics, Munich (Germany); Arai, S. [Tokyo University of Agriculture and Technology, Tokyo (Japan); Arogancia, D.C.; Bacala, A.M. [Mindanao State University, Iligan City (Philippines); Ball, M.; Behnke, T. [DESY, Hamburg (Germany); Bito, H. [Tokyo University of Agriculture and Technology, Tokyo (Japan); Eckardt, V. [Max-Planck-Institute for Physics, Munich (Germany); Fujii, K. [KEK, IPNS, Ibaraki (Japan); Fusayasu, T. [Nagasaki Institute of Applied Science, Nagasaki (Japan); Ghodbane, N. [DESY, Hamburg (Germany); Gooc, H.C Jr. [Mindanao State University, Iligan City (Philippines); Kijima, T. [Tokyo University of Agriculture and Technology, Tokyo (Japan); Hamann, M. [DESY, Hamburg (Germany); Habu, M. [Tokyo University of Agriculture and Technology, Tokyo (Japan); Heuer, R.-D. [Universitaet Hamburg, Hamburg (Germany); Hiramatsu, K. [Kinki University, Osaka (Japan); Ikematsu, K. [KEK, IPNS, Ibaraki (Japan); Kaukher, A. [Universitaet Rostock, Rostock (Germany); Kuroiwa, H. [Saga University, Saga (Japan)

    2010-11-01

    A time projection chamber (TPC) is a strong candidate for the central tracker of the international linear collider (ILC) experiment and we have been conducting a series of cosmic ray experiments under a magnetic field up to 4 T, using a small prototype TPC with a replaceable readout device: multi-wire proportional chamber (MWPC) or gas electron multiplier (GEM). We first confirmed that the MWPC readout could not be a fall-back option of the ILC-TPC under a strong axial magnetic field of 4 T since its spatial resolution suffered severely from the so called ExB effect in the vicinity of the wire planes. The GEM readout, on the other hand, was found to be virtually free from the ExB effect and gave the resolution determined by the transverse diffusion of the drift electrons (diffusion limited). Furthermore, GEMs allow a wider choice of gas mixtures than MWPCs. Among the gases we tried so far a mixture of Ar-CF{sub 4}-isobutane seems promising as the operating gas of the ILC-TPC because of its small diffusion constant especially under a strong magnetic field. We report the spatial resolution obtained with the GEM readout in this gas mixture. Also presented is the spatial resolution of a GEM-based ILC-TPC estimated from the measurement with the prototype.

  15. Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

    Directory of Open Access Journals (Sweden)

    Heger Christopher D

    2010-12-01

    Full Text Available Abstract Background Prolactin is a polypeptide hormone responsible for proliferation and differentiation of the mammary gland. More recently, prolactin's role in mammary carcinogenesis has been studied with greater interest. Studies from our laboratory and from others have demonstrated that three specific isoforms of the prolactin receptor (PRLR are expressed in both normal and cancerous breast cells and tissues. Until now, reliable isoform specific antibodies have been lacking. We have prepared and characterized polyclonal antibodies against each of the human PRLR isoforms that can effectively be used to characterize human breast cancers. Methods Rabbits were immunized with synthetic peptides of isoform unique regions and immune sera affinity purified prior to validation by Western blot and immunohistochemical analyses. Sections of ductal and lobular carcinomas were stained with each affinity purified isoform specific antibody to determine expression patterns in breast cancer subclasses. Results We show that the rabbit antibodies have high titer and could specifically recognize each isoform of PRLR. Differences in PRLR isoform expression levels were observed and quantified using histosections from xenografts of established human breast cancer cells lines, and ductal and lobular carcinoma human biopsy specimens. In addition, these results were verified by real-time PCR with isoform specific primers. While nearly all tumors contained LF and SF1b, the majority (76% of ductal carcinoma biopsies expressed SF1a while the majority of lobular carcinomas lacked SF1a staining (72% and 27% had only low levels of expression. Conclusions Differences in the receptor isoform expression profiles may be critical to understanding the role of PRL in mammary tumorigenesis. Since these antibodies are specifically directed against each PRLR isoform, they are valuable tools for the evaluation of breast cancer PRLR content and have potential clinical importance in

  16. Noise Filtering and Signal Calibration in the MicroBooNE LArTPC

    Science.gov (United States)

    Joshi, Jyoti; Qian, Xin; MicroBooNE Collaboration Collaboration

    2017-01-01

    In large liquid argon time projection chambers (LArTPCs), TPC signal processing, which recovers the number of ionized electrons arriving at anode wire planes from the raw digitized induction signals, is a crucial step towards automated event reconstruction. The first stage of signal processing is the identification and removal of any excess TPC noise with minimal impact on the true signal. In this talk, first I will describe the characterization and software filtering techniques of various TPC noise observed in the raw digital signal data in MicroBooNE. I will then describe a novel drifted-charge extraction method based on 2D deconvolution technique. These techniques significantly enhance the performance of the induction wire planes in MicroBooNE.

  17. TPC track distortions: correction maps for magnetic and static electric inhomogeneities

    CERN Document Server

    Dydak, F; Nefedov, Y

    2003-01-01

    Inhomogeneities of the magnetic and electric fields in the active TPC volume lead to displacements of cluster coordinates, and therefore to track distortions. In case of good data taking conditions, the largest effects are expected from the inhomogeneity of the solenoidal magnetic field, and from a distortion of the electric field arising from a high voltage misalignment between the outer and inner field cages. Both effects are stable over the entire HARP data taking. The displacements are large compared to the azimuthal coordinate resolution but can be corrected with sufficient precision, except at small TPC radius. The high voltage misalignment between the outer and inner field cages is identified as the likely primary cause of sagitta distortions of TPC tracks. The position and the length of the target plays an important role. Based on a detailed modelling of the magnetic and static electric field inhomogeneities, precise correction maps for both effects have been calculated. Predictions from the correctio...

  18. The NEXT experiment: A high pressure xenon gas TPC for neutrinoless double beta decay searches

    CERN Document Server

    Lorca, D; Monrabal, F

    2012-01-01

    Neutrinoless double beta decay is a hypothetical, very slow nuclear transition in which two neutrons undergo beta decay simultaneously and without the emission of neutrinos. The importance of this process goes beyond its intrinsic interest: an unambiguous observation would establish a Majorana nature for the neutrino and prove the violation of lepton number. NEXT is a new experiment to search for neutrinoless double beta decay using a radiopure high-pressure xenon gas TPC, filled with 100 kg of Xe enriched in Xe-136. NEXT will be the first large high-pressure gas TPC to use electroluminescence readout with SOFT (Separated, Optimized FuncTions) technology. The design consists in asymmetric TPC, with photomultipliers behind a transparent cathode and position-sensitive light pixels behind the anode. The experiment is approved to start data taking at the Laboratorio Subterr\\'aneo de Canfranc (LSC), Spain, in 2014.

  19. Design of a Radial TPC for Antihydrogen Gravity Measurement with ALPHA-g

    CERN Document Server

    Capra, Andrea; Bishop, Daryl; Fujiwara, Makoto C.; Freeman, Skyler; Gill, David; Grant, Matthew; Henderson, Robert; Kurchaninov, Leonid; Lu, Philip; Menary, Scott; Olchanski, Konstantin; Retiere, Fabrice

    2016-01-01

    The gravitational interaction of antimatter and matter has never been directly probed. ALPHA-g is a novel experiment that aims to perform the first measurement of the antihydrogen gravitational mass. A fundamental requirement for this new apparatus is a position sensitive particle detector around the antihydrogen trap which provides information about antihydrogen annihilation location. The proposed detector is a radial Time Projection Chamber, or \\textit{rTPC}, whose concept is being developed at TRIUMF. A simulation of the detector and the development of the reconstruction software, used to determine the antihydrogen annihilation point, is presented alongside with the expected performance of the rTPC.

  20. TPC-BASED STBC MULTIUSER DETECTION WITH LSE-RLS ALGORITHM

    Institute of Scientific and Technical Information of China (English)

    Du Yinggang; Chan Kam Tai

    2006-01-01

    The Bit Error Rate (BER) performance of a Turbo Product Code (TPC) based Space-Time Block Coding (STBC) multiuser wireless system in the frequency-selective channels has been investigated. Both of the good error correcting capability of TPC and the large diversity gain of STBC can be achieved simultaneously. A Least Square Error-Recursive Least Square (LSE-RLS) algorithm is applied to estimate the channel and cancel the interference. Simulations show that the proposed system can obtain about 2.7dB gain in Es/No at the BER of 10-3.

  1. An optical readout TPC (O-TPC) for studies in nuclear astrophysics with gamma-ray beams at HI{gamma}S{sup 1}

    Energy Technology Data Exchange (ETDEWEB)

    Gai, M; Zimmerman, W R; Kading, T J; Seo, P-N; Young, A H [LNS at Avery Point, University of Connecticut, Groton, CT 06340-6097 (United States); Ahmed, M W; Stave, S C; Henshaw, S S; Martel, P P; Weller, H R [TUNL, Dept. of Physics, Duke University, Durham, NC 27708 (United States); Breskin, A; Chechik, R [Dept. of Particle Physics, Weizmann Institute of Science, 76100 Rehovot (Israel); Bromberger, B; Dangendorf, V; Tittelmeier, K [Physikalisch-Technische Bundesanstalt, 38116 Braunschweig (Germany); Delbar, Th [Universite Catholique de Louvain, 1348 Louvain-la-Neuve (Belgium); III, R H France [Georgia College and State University, CBX 82, Milledgeville, GA 31061 (United States); McDonald, J E R, E-mail: moshe.gai@yale.edu [Dept. of Physics, Yale University, New Haven, CT 06520-8124 (United States)

    2010-12-15

    We report on the construction, tests, calibrations and commissioning of an Optical Readout Time Projection Chamber (O-TPC) detector operating with a CO{sub 2}(80%) + N{sub 2}(20%) gas mixture at 100 and 150 Torr. It was designed to measure the cross sections of several key nuclear reactions involved in stellar evolution. In particular, a study of the rate of formation of oxygen and carbon during the process of helium burning will be performed by exposing the chamber gas to intense nearly mono-energetic gamma-ray beams at the High Intensity Gamma Source (HI{gamma}S) facility. The O-TPC has a sensitive target-drift volume of 30x30x21 cm{sup 3}. Ionization electrons drift towards a double parallel-grid avalanche multiplier, yielding charge multiplication and light emission. Avalanche-induced photons from N{sub 2} emission are collected, intensified and recorded with a Charge Coupled Device (CCD) camera, providing two-dimensional track images. The event's time projection (third coordinate) and the deposited energy are recorded by photomultipliers and by the TPC charge-signal, respectively. A dedicated VME-based data acquisition system and associated data analysis tools were developed to record and analyze these data. The O-TPC has been tested and calibrated with 3.183 MeV alpha-particles emitted by a {sup 148}Gd source placed within its volume with a measured energy resolution of 3.0%. Tracks of alpha and {sup 12}C particles from the dissociation of {sup 16}O and of three alpha-particles from the dissociation of {sup 12}C have been measured during initial in-beam test experiments performed at the HI{gamma}S facility at Duke University. The full detection system and its performance are described and the results of the preliminary in-beam test experiments are reported.

  2. Measurement of the drift field in the ARGONTUBE LAr TPC with 266~nm pulsed laser beams

    CERN Document Server

    Ereditato, A; Janos, S; Kreslo, I; Luethi, M; von Rohr, C Rudolf; Schenk, M; Strauss, T; Weber, M S; Zeller, M

    2014-01-01

    ARGONTUBE is a liquid argon time projection chamber (LAr TPC) with a drift field generated in-situ by a Greinacher voltage multiplier circuit. We present results on the measurement of the drift field distribution based on a simplified model of a multi-stage Greinacher circuit, by using straight ionization tracks generated by an intense UV laser beam.

  3. Measuring Cross-Section and Estimating Uncertainties with the fissionTPC

    Energy Technology Data Exchange (ETDEWEB)

    Bowden, N. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Manning, B. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Sangiorgio, S. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Seilhan, B. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2015-01-30

    The purpose of this document is to outline the prescription for measuring fission cross-sections with the NIFFTE fissionTPC and estimating the associated uncertainties. As such it will serve as a work planning guide for NIFFTE collaboration members and facilitate clear communication of the procedures used to the broader community.

  4. Signal Processing in the MicroBooNE LArTPC

    CERN Document Server

    Joshi, Jyoti

    2015-01-01

    The MicroBooNE experiment is designed to observe interactions of neutrinos with a Liquid Argon Time Projection Chamber (LArTPC) detector from the on-axis Booster Neutrino Beam (BNB) and off-axis Neutrinos at the Main Injector (NuMI) beam at Fermi National Accelerator Laboratory. The detector consists of a $2.5~m\\times 2.3~m\\times 10.4~m$ TPC including an array of 32 PMTs used for triggering and timing purposes. The TPC is housed in an evacuable and foam insulated cryostat vessel. It has a 2.5 m drift length in a uniform field up to 500 V/cm. There are 3 readout wire planes (U, V and Y co-ordinates) with a 3-mm wire pitch for a total of 8,256 signal channels. The fiducial mass of the detector is 60 metric tons of LAr. In a LArTPC, ionization electrons from a charged particle track drift along the electric field lines to the detection wire planes inducing bipolar signals on the U and V (induction) planes, and a unipolar signal collected on the (collection) Y plane. The raw wire signals are processed by speciali...

  5. Lessons from the operation of the `Penning-Fluorescent' TPC and prospects

    CERN Document Server

    Gonzalez-Diaz, Diego; Castel, J; Cebrian, S; Dafni, T; Garcia, J A; Gomez, H; Herrera, D C; Iguaz, F J; Irastorza, I G; Lagraba, A; Luzon, G; Rodriguez, A; Ruiz-Choliz, E; Ferrer-Ribas, A Tomas E; Giomataris, I

    2015-01-01

    We have recently reported the development of a new type of high-pressure Xenon time projection chamber operated with an ultra-low diffusion mixture and that simultaneously displays Penning effect and fluorescence in the near-visible region (300 nm). The concept, dubbed `Penning-Fluorescent' TPC, allows the simultaneous reconstruction of primary charge and scintillation with high topological and calorimetric fidelity.

  6. Particle Identification in Jets and High-Multiplicity pp Events with the ALICE TPC

    CERN Document Server

    AUTHOR|(SzGeCERN)683272; Vogelsang, Werner

    The spectra of identified particles in a collision experiment comprise crucial information about the underlying physical processes. The ALICE experiment has powerful Particle IDentification (PID) capabilities, which are unique at the Large Hadron Collider (LHC). In this thesis, a statistical PID method based on the specific energy loss d$E$/d$x$ in the ALICE Time Projection Chamber (TPC) is developed: the TPC Multi-Template Fit (MTF). The MTF allows for the extraction of identified charged particle spectra in a wide momentum range, which extends from about 150 MeV/$c$ to above 20 GeV/$c$. The TPC PID requires a detailed modelling of the TPC d$E$/d$x$ response for momenta above 2-3 GeV/$c$. A framework is developed that allows for the determination of the model parameters and for evaluating the PID information of charged particles. With the MTF, the transverse momentum $p_{\\mathrm{T}}$ spectra of charged pions, kaons and protons at mid-rapidity ($|\\eta| < 0.9$) are measured for pp collisions at $\\sqrt{s}$ ...

  7. Study of electron recombination in liquid argon with the ICARUS TPC

    Energy Technology Data Exchange (ETDEWEB)

    Amoruso, S.; Antonello, M.; Aprili, P.; Arneodo, F.; Badertscher, A.; Baiboussinov, B.; Baldo Ceolin, M.; Battistoni, G.; Bekman, B.; Benetti, P.; Bischofberger, M.; Borio di Tigliole, A.; Brunetti, R.; Bruzzese, R.; Bueno, A.; Buzzanca, M.; Calligarich, E.; Campanelli, M.; Carbonara, F.; Carpanese, C.; Cavalli, D.; Cavanna, F.; Cennini, P.; Centro, S.; Cesana, A.; Chen, C.; Chen, D.; Chen, D.B.; Chen, Y.; Cieslik, K.; Cline, D.; Cocco, A.G.; Dai, Z.; De Vecchi, C.; Dabrowska, A.; Di Cicco, A.; Dolfini, R.; Ereditato, A.; Felcini, M.; Ferrari, A.; Ferri, F.; Fiorillo, G.; Galli, S.; Ge, Y.; Gibin, D.; Gigli Berzolari, A.; Gil-Botella, I.; Graczyk, K.; Grandi, L.; Guglielmi, A.; He, K.; Holeczek, J.; Huang, X.; Juszczak, C.; Kielczewska, D.; Kisiel, J.; Kozlowski, T.; Laffranchi, M.; Lagoda, J.; Li, Z.; Lu, F.; Ma, J.; Mangano, G.; Markiewicz, M.; Martinez de la Ossa, A.; Matthey, C.; Mauri, F.; Meng, G.; Messina, M.; Montanari, C.; Muraro, S.; Navas-Concha, S.; Otwinowski, S.; Ouyang, Q.; Palamara, O.; Pascoli, D.; Periale, L.; Piano Mortari, G.B.; Piazzoli, A.; Picchi, P.; Pietropaolo, F.; Polopek, W.; Rancati, T.; Rappoldi, A.; Raselli, G.L.; Rico, J.; Rondio, E.; Rossella, M.; Rubbia, A.; Rubbia, C.; Sala, P.R. E-mail: paola.sala@cern.ch; Santorelli, R.; Scannicchio, D.; Segreto, E.; Seo, Y.; Sergiampietri, F.; Sobczyk, J.; Spinelli, N.; Stepaniak, J.; Sulej, R.; Szarska, M.; Szeptycka, M.; Terrani, M.; Velotta, R.; Ventura, S.; Vignoli, C.; Wang, H.; Wang, X.; Woo, J.; Xu, G.; Xu, Z.; Zalewska, A.; Zhang, C.; Zhang, Q.; Zhen, S.; Zipper, W

    2004-05-11

    Electron recombination in liquid argon (LAr) is studied by means of charged particle tracks collected in various ICARUS liquid argon TPC prototypes. The dependence of the recombination on the particle stopping power has been fitted with a Birks functional dependence. The simulation of the process of electron recombination in Monte Carlo calculations is discussed. A quantitative comparison with previously published data is carried out.

  8. Front-end electronics and readout system for the ILD TPC

    CERN Document Server

    Hedberg, V; Lundberg, B; Mjörnmark, U; Oskarsson, A; Österman, L; De Lentdecker, G; Yang, Y; Zhang, F

    2015-01-01

    A high resolution TPC is the main option for a central tracking detector at the future International Linear Collider (ILC). It is planned that the MPGD (Micro Pattern Gas Detector) technology will be used for the readout. A Large Prototype TPC at DESY has been used to test the performance of MPGDs in an electron beam of energies up to 6 GeV. The first step in the technology development was to demonstrate that the MPGDs are able to achieve the necessary performance set by the goals of ILC. For this ’proof of principle’ phase, the ALTRO front-end electronics from the ALICE TPC was used, modified to adapt to MPGD readout. The proof of principle has been verified and at present further improvement of the MPGD technology is going on, using the same readout electronics. The next step is the ’feasibility phase’, which aims at producing front-end electronics comparable in size (few mm2) to the readout pads of the TPC. This development work is based on the succeeding SALTRO16 chip, which combines the analogue ...

  9. Process for purifying geothermal steam

    Science.gov (United States)

    Li, C.T.

    Steam containing hydrogen sulfide is purified and sulfur recovered by passing the steam through a reactor packed with activated carbon in the presence of a stoichiometric amount of oxygen which oxidizes the hydrogen sulfide to elemental sulfur which is adsorbed on the bed. The carbon can be recycled after the sulfur has been recovered by vacuum distillation, inert gas entrainment or solvent extraction. The process is suitable for the purification of steam from geothermal sources which may also contain other noncondensable gases.

  10. Conotoxins Are Purified and Cloned

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    @@ A group of CAS scientists have succeeded in purifying many conotoxins and cloning more than 100 new genes from six species of cone snails living in waters off the coast of the South China Sea, paving the way for the development of new drugs to relieve neuropathic pains. The work has been honored with a first prize from the 2005 Awards for S&T Progress in Shanghai.

  11. Design of a neutron-TPC prototype and its performance evaluation based on an alpha-particle test

    Science.gov (United States)

    Huang, Meng; Li, Yu-Lan; Niu, Li-Bo; Li, Jin; Deng, Zhi; He, Li; Zhang, Hong-Yan; Cheng, Xiao-Lei; Fu, Jian-Qiang; Li, Yuan-Jing

    2015-08-01

    A neutron-TPC (nTPC) is being developed for use as a fast neutron spectrometer in the fields of nuclear physics, nuclear reactor operation monitoring, and thermo-nuclear fusion plasma diagnostics. An nTPC prototype based on a GEM-TPC (Time Projection Chamber with Gas Electron Multiplier amplification) has been assembled and tested using argon-hydrocarbon mixture as the working gas. By measuring the energy deposition of the recoil proton in the sensitive volume and the angle of the proton track, the incident neutron energy can be deduced. A Monte Carlo simulation was carried out to analyze the parameters affecting the energy resolution of the nTPC, and gave an optimized resolution under ideal conditions. An alpha particle experiment was performed to verify its feasibility, and to characterize its performance, including energy resolution and spatial resolution. Based on the experimental measurement and analysis, the energy resolution (FWHM) of the nTPC prototype is predicted to be better than 3.2% for 5 MeV incident neutrons, meeting the performance requirement (FWHM<5%) for the nTPC prototype.

  12. Protein chemical characterization of Gc globulin (vitamin D-binding protein) isoforms; Gc-1f, Gc-1s and Gc-2

    DEFF Research Database (Denmark)

    Christiansen, Maja; Jørgensen, Charlotte S; Laursen, Inga;

    2007-01-01

    exchange chromatography. The separated isoforms and several commercial preparations of individual isoforms were characterized by mass spectrometry. This revealed that the major isoforms were non-glycosylated. Compared to the Gc-1f isoform the other dominating isoforms represented an Asp/Glu substitution......Gc globulin, also called vitamin D-binding protein, is a plasma protein involved in the extracellular actin-scavenger system, vitamin D transport and possibly also other biological activities. Low levels of Gc globulin have been found to correlate with multiple organ failure and non......-survival of patients with fulminant hepatic failure and trauma. Here, we characterize the dominant isoforms of plasma-derived Gc globulin from Cohn fraction IV paste with respect to amino acid sequence and posttranslational modifications. Gc globulin was purified in large scale and the isoforms separated by ion...

  13. Histamine H3-receptor isoforms.

    Science.gov (United States)

    Bakker, R A

    2004-10-01

    Increasing evidence supports a role for HA as a neurotransmitter and neuromodulator in various brain functions, including emotion, cognition, and feeding. The recent cloning of the histamine H3 receptor allowed for the subsequent cloning of a variety of H3 receptor isoforms from different species as well as the H4 receptor. As a result a wide variety of H3-receptor isoforms are now known that display differential brain expression patterns and signalling properties. These recent discoveries are discussed in view of the growing interest of the H3 receptor as a target for the development of potential therapeutics.

  14. Purification and characterization of soluble (cytosolic) and bound (cell wall) isoforms of invertases in barley (Hordeum vulgare) elongating stem tissue

    Science.gov (United States)

    Karuppiah, N.; Vadlamudi, B.; Kaufman, P. B.

    1989-01-01

    Three different isoforms of invertases have been detected in the developing internodes of barley (Hordeum vulgare). Based on substrate specificities, the isoforms have been identified to be invertases (beta-fructosidases EC 3.2.1.26). The soluble (cytosolic) invertase isoform can be purified to apparent homogeneity by diethylaminoethyl cellulose, Concanavalin-A Sepharose, organo-mercurial Sepharose, and Sephacryl S-300 chromatography. A bound (cell wall) invertase isoform can be released by 1 molar salt and purified further by the same procedures as above except omitting the organo-mercurial Sepharose affinity chromatography step. A third isoform of invertase, which is apparently tightly associated with the cell wall, cannot be isolated yet. The soluble and bound invertase isoforms were purified by factors of 60- and 7-fold, respectively. The native enzymes have an apparent molecular weight of 120 kilodaltons as estimated by gel filtration. They have been identified to be dimers under denaturing and nondenaturing conditions. The soluble enzyme has a pH optimum of 5.5, Km of 12 millimolar, and a Vmax of 80 micromole per minute per milligram of protein compared with cell wall isozyme which has a pH optimum of 4.5, Km of millimolar, and a Vmax of 9 micromole per minute per milligram of protein.

  15. Methods for Purifying Enzymes for Mycoremediation

    Science.gov (United States)

    Cullings, Kenneth W. (Inventor); DeSimone, Julia C. (Inventor); Paavola, Chad D. (Inventor)

    2014-01-01

    A process for purifying laccase from an ectomycorrhizal fruiting body is disclosed. The process includes steps of homogenization, sonication, centrifugation, filtration, affinity chromatography, ion exchange chromatography, and gel filtration. Purified laccase can also be separated into isomers.

  16. Evaluation of a portable air purifier.

    OpenAIRE

    Lawrence, J.C.; Lilly, H. A.; Wilkins, M. D.

    1981-01-01

    A portable air purifier significantly reduced mal odour in a small room. If the atmosphere was deliberately contaminated with Serratia marcescens the unit rapidly removed this organism. However, if incorrectly sited, the purifier could disperse organisms into the atmosphere.

  17. Energy calibration of a GEM-TPC prototype with 83mKr

    Science.gov (United States)

    Schmitz, Roman

    2012-05-01

    A GEM-TPC has been built as a 10% scale prototype for the P¯ANDA experiment and as a full-scale prototype for the CBELSA/TAPS experiment (Fabbietti L, et al: Nucl Instrum Methods A 628:204, 2011). The intrinsic suppression of ion backflow into the drift volume makes it suitable for high rate/background environments. The GEM-TPC has been used as an inner tracking update for the FOPI experiment at GSI where cosmic and beam tests were performed. For calibration purposes a 83Rb source has been produced at the Bonn HISKP Cyclotron. Using the isomeric 83mKr evaporated by the source, a relative channel-wise gain calibration as well as gain estimation for different high voltage settings could be performed.

  18. On distortions of TPC coordinates: inhomogeneities of electric and magnetic field

    CERN Document Server

    Dydak, F

    2003-01-01

    After a general discussion of electron drift in a gas volume with electric and magnetic fields, distortions in the r and r phi coordinates arising from inhomogeneities of the electric and magnetic fields in the HARP TPC are calculated. Inhomogeneities of the electric field arise from i) positive ions released by cosmic rays, ii) positive ions released by interaction secondaries, iii) positive ions released by beam muons, iv) positive ions released from beam particles downstream of the inner field cage, and v) a high voltage misalignment between the outer and inner field cages. Also, distortions arising from the inhomogeneity of the magnetic field are calculated. These effects resolve the controversy on unphysical numbers of 'wrong-charge' TPC tracks. The bad news are that effects are too big to be neglected. The good news are that, with enough sweat and tears, they can be adequately corrected.

  19. Tests of gases in a mini-TPC with pixel chip readout

    Energy Technology Data Exchange (ETDEWEB)

    Vahsen, S. [University of Hawaii, 2505 Correa Road, Honolulu, HI 96822 (United States); Oliver-Mallory, K.; Lopez-Thibodeaux, M. [Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States); Kadyk, J., E-mail: jakadyk@lbl.gov [Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States); Garcia-Sciveres, M. [Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States)

    2014-02-21

    Gases for potential use as targets for directional dark matter detection were tested in a prototype detector using two sequential Gas Electron Multipliers, or GEMs. The sensitive volume consists of a mini-TPC of 12 cm length and 7.5 cm diameter. An FEI3 pixel chip, developed for the ATLAS experiment, was used to produce spatial measurements with high resolution. An Fe55 source produced photoelectrons by X-ray conversions in the sensitive volume, and images of these were recorded by the chip. Spatial resolution plots are shown for the gases, which include the practical electron range of the photoelectrons and the effects of diffusion in the mini-TPC. Avalanche gain and gain resolution measurements were made for the four gases tested, at atmospheric and sub-atmospheric pressures: Ar(70)/CO{sub 2}(30), CF{sub 4}, He(80)/CF{sub 4}(20) and He(80)/isobutane(20)

  20. Magnetic Field Requirements for a Detector at the Linear Collider Using a TPC as Main Tracking Device

    CERN Document Server

    Klempt, W

    2010-01-01

    This note describes the requirements to the magnetic field which occur in an ILD like detector at ILC or CLIC. In particular we describe requirements introduced by choosing a TPC as main tracking detector.

  1. TPB-coated Light Guides for Liquid Argon TPC Light Detection Systems

    CERN Document Server

    Ignarra, C M

    2013-01-01

    Light detection systems in Liquid Argon Time Projection Chambers (LArTPCs) require the detection of the 128 nm light produced during argon scintillation. Most detectors use Tetraphenyl Butadiene (TPB) to shift the wavelength of the light into a range visible to Photomultiplier Tubes (PMTs). These proceedings summarize characterizations of light-guides coated with a matrix of TPB in UV transmitting acrylic which are more compact than existing LArTPC light collection systems.

  2. Simulation of double beta decay in the ''SeXe'' TPC

    Science.gov (United States)

    Mauger, F.

    2007-04-01

    In 2004, the NEMO collaboration has started some preliminary studies for a next-generation double beta decay experiment: SuperNEMO. The possibility to use a large gaseous TPC has been investigated using simulation and extrapolation of former experiments. In this talk, I report on the reasons why such techniques have not been selected in 2004 and led the NEMO collaboration to reuse the techniques implemented within the NEMO3 detector.

  3. Simulation of double beta decay in the 'SeXe' TPC

    Energy Technology Data Exchange (ETDEWEB)

    Mauger, F [LPC Caen and University of Caen, ENSICAEN, 6 Bd Marechal Juin, 14050 CAEN CEDEX 4 (France)

    2007-04-15

    In 2004, the NEMO collaboration has started some preliminary studies for a next-generation double beta decay experiment: SuperNEMO. The possibility to use a large gaseous TPC has been investigated using simulation and extrapolation of former experiments. In this talk, I report on the reasons why such techniques have not been selected in 2004 and led the NEMO collaboration to reuse the techniques implemented within the NEMO3 detector.

  4. Electron lifetime measurement using cosmic ray muons at the MicroBooNE LArTPC

    Science.gov (United States)

    Meddage, Varuna Crishan; MicroBooNE Collaboration

    2017-01-01

    MicroBooNE, a 170 ton liquid argon time projection chamber (LArTPC) located on the Fermilab's Booster Neutrino Beamline (BNB), is designed to both probe neutrino physics phenomena and further develop the LArTPC detector technology. MicroBooNE is the largest currently operating LArTPC detector and began collecting data in Fall 2015. LArTPCs are imaging detectors that offer exceptional capabilities for studying neutrinos. A fundamental requirement for the performance of such detectors is to maintain electronegative contaminants such as oxygen and water at extremely low concentrations, which otherwise can absorb the ionization electrons. The impurity levels in liquid argon can be estimated from the drift electron lifetime as they are inversely proportional to each other. This talk presents a measurement of the drift electron lifetime using cosmic ray muon data collected by MicroBooNE. An interpretation of the observed drift electron lifetime as a function of time indicates that the electron attenuation due to impurities in the liquid argon is negligible during normal operations, implying that the argon purification and gas recirculation system in MicroBooNE is performing successfully.

  5. Search for H-dibaryon at J-PARC with a Large Acceptance TPC

    Directory of Open Access Journals (Sweden)

    Sako H.

    2014-03-01

    Full Text Available H-dibaryon has been predicted as a stable 6-quark color-singlet state. It has been searched for by many experiments but has never been discovered. Recent lattice QCD calculations predict H-dibaryon as a weakly bound or a resonant state close to the LL threshold. E224 and E522 experiments at KEK observed peaks in LL invariant mass spectra near the threshold in (K-, K+ reactions, which were statistically not significant. Therefore, we proposed a new experiment E42 at J-PARC. It will measure decay products of ΛΛ and Λπ-p in a (K-, K+ reaction. We design a large acceptance spectrometer based on a Time Projection Chamber (TPC immersed in a dipole magnetic field. The TPC surrounds a target to cover nearly 4π acceptance, and accepts K- beams up to 106 counts per second. To suppress drift field distortion at high beam rates, we adopt Gas Electron Multipliers (GEMs for electron amplification and a gating grid. We show an overview of the experiment, the design of the spectrometer, and the R&D status of the TPC prototype.

  6. Optical Readout of a Two Phase Liquid Argon TPC using CCD Camera and TGEMs

    CERN Document Server

    Mavrokoridis, K; Carroll, J; Lazos, M; McCormick, K J; Smith, N A; Touramanis, C; Walker, J

    2014-01-01

    This paper presents a preliminary study into the use of CCDs to image secondary scintillation light generated by Thick Gas Electron Multipliers (TGEMs) in a two phase LAr TPC. A Sony ICX285AL CCD chip was mounted above a double TGEM in the gas phase of a 40 litre two-phase LAr TPC with the majority of the camera electronics positioned externally via a feedthrough. An Am-241 source was mounted on a rotatable motion feedthrough allowing the positioning of the alpha source either inside or outside of the field cage. Developed for and incorporated into the TPC design was a novel high voltage feedthrough featuring LAr insulation. Furthermore, a range of webcams were tested for operation in cryogenics as an internal detector monitoring tool. Of the range of webcams tested the Microsoft HD-3000 (model no:1456) webcam was found to be superior in terms of noise and lowest operating temperature. In ambient temperature and atmospheric pressure 1 ppm pure argon gas, the TGEM gain was approximately 1000 and using a 1 msec...

  7. Status of the R&D activities for the upgrade of the ALICE TPC

    CERN Document Server

    Deisting, Alexander

    2016-01-01

    In order to cope with the high interaction rates provided by the LHC after the long shut-down 2, the ALICE TPC needs to be upgraded. After the upgrade the TPC will run in a continuous mode, without any degradation of the momentum and $\\textrm{d}E/\\textrm{d}x$ resolution compared to the performance of the present TPC. Since readout by MWPCs is no longer feasible with these requirements, new technologies have to be employed. In the new readout the electron amplification is provided by a stack of four GEM foils. Here foils with a standard hole pitch of 140um as well as large pitch foils (280um) are used. Their high voltage settings and orientation have been optimised to provide an energy resolution of $\\sigma_{{E}}/{E}\\leq12\\%$ at the photopeak of $^{55}\\textrm{Fe}$. At the same settings the Ion BackFlow into the drift volume is less than 1% of the effective number of ions produced during gas amplification and the primary ionisations. This is necessary to prevent the accumulation of space charge, which eventuall...

  8. Simulation and Calibration of the ALICE TPC including innovative Space Charge Calculations

    CERN Document Server

    Rossegger, S; Riegler, W; Betev, L

    2009-01-01

    ALICE is one of the four main particle detectors located around the LHC accelerator at CERN. It is particularly designed to study the physics of the quark-gluon plasma by means of nucleus--nucleus collisions at center-of-mass energies up to 5.5 TeV per nucleon pair. A Time-Projection Chamber (TPC) was chosen to be its central-sub-detector due to its low mass properties and its capabilities to provide a robust and accurate Particle Identification even within ultra-high multiplicity environments (up to 8000 tracks per unit of eta). To achieve the required physics performance, the space point resolution of the TPC must be in the order of 0.2 mm. Due to its gigantic size of 5~m in diameter and 5~m in length, corrections for static as well as dynamic effects are indispensable in order to accomplish the design goal. The research presented covers all major issues relevant for the final calibration and therefore the enhancement of the TPC performance in terms of resolution. The main focus was to distinguish between t...

  9. Gamma-ray imaging with a large micro-TPC and a scintillation camera

    Energy Technology Data Exchange (ETDEWEB)

    Hattori, K. [Graduate School of Science, Department of Physics, Kyoto University Kitashirakawa, Sakyo, Kyoto 606-8502 (Japan)], E-mail: hattori@cr.scphys.kyoto-u.ac.jp; Kabuki, S.; Kubo, H.; Kurosawa, S.; Miuchi, K. [Graduate School of Science, Department of Physics, Kyoto University Kitashirakawa, Sakyo, Kyoto 606-8502 (Japan); Nagayoshi, T. [Advanced Research Institute for Science and Engineering, Waseda University, 17 Kikui-cho, Shinjuku 162-0044, Tokyo (Japan); Nishimura, H.; Okada, Y. [Graduate School of Science, Department of Physics, Kyoto University Kitashirakawa, Sakyo, Kyoto 606-8502 (Japan); Orito, R. [Graduate School of Science and Technology, Department of Physics, Kobe University, 1-1 Rokkoudai, Nada, Kobe 657-8501 (Japan); Sekiya, H.; Takada, A. [Graduate School of Science, Department of Physics, Kyoto University Kitashirakawa, Sakyo, Kyoto 606-8502 (Japan); Takeda, A. [Kamioka Observatory, ICRR, University of Tokyo, 456 Higashi-mozumi, Hida-shi, Gifu 505-1205 (Japan); Tanimori, T.; Ueno, K. [Graduate School of Science, Department of Physics, Kyoto University Kitashirakawa, Sakyo, Kyoto 606-8502 (Japan)

    2007-10-21

    We report on the development of a large Compton camera with the full reconstruction of the Compton process based on a prototype. This camera consists of two kinds of detectors. One is a gaseous time projection chamber (micro-TPC) for measuring the energy and the track of a Compton recoil electron. The micro-TPC is based on a {mu}-PIC and a GEM, which are micro-pattern gas detectors (MPGDs). The size of the micro-TPC was 10cmx10cmx8cm in the case of the prototype, and we enlarged it to 23cmx28cmx15cm. The other detector part is a NaI (Tl) Anger camera for measuring the scattered gamma-ray. With these informations, we can completely reconstruct a Compton event, and determine the direction of the incident gamma-ray, event by event. We succeeded in reconstructing events of incident 662 keV gamma-rays. The measured angular resolutions of the 'angular resolution measure' (ARM) and the 'scatter plane deviation' (SPD) were 9.3{sup 0} and 158{sup 0} (FWHM), respectively.

  10. AGS silicon gold collisions measured in the E-810 TPC (Time Projection Chamber)

    Energy Technology Data Exchange (ETDEWEB)

    Bonner, B.E.; Buchanan, J.A.; Chiou, C.N.; Clement, J.M.; Corcoran, M.D.; Kruk, J.W.; Miettinen, H.E.; Mutchler, G.S.; Nessi, M.; Nessi-Tedaldi, F.; Roberts, J.B. (Rice Univ., Houston, TX (USA)); Chan, C.S.; Kramer, M.A. (City Coll., New York, NY (USA)); Etkin, A.; Foley, K.J.; Hackenburg, R.W.; Longacre, R.S.; Love, W.A.; Morris, T.W.; Platner, E.D.; Saulys, A.C. (Brookhaven National Lab., Upton, NY (USA)); Hallma

    1990-03-26

    The tracking detector of AGS Experiment 810 is a three-piece Time Projection Chamber (TPC) intended to measure all charged tracks in the forward hemisphere of the nucleon-nucleon center of mass system, i.e. forward of an angle of about 20 degrees in the lab. Each module of the TPC contains twelve rows of short anode wires which give 3-D space points on each track, but no dE/dx information useable for particle identification. The TPC was operated in a beam of silicon ions at the end of June 1989 and this talk reports the results of analysis of the data taken with a thin gold target in that run. We have gathered a similar amount of data from thin copper and silicon targets, the analysis of which is in a less advanced state. The results of our investigation of the neutral strange particle decays appear in a separate contribution by Al Saulys. This paper presents the current state of the analysis of the charged tracks from the silicon gold collisions. 1 ref., 15 figs.

  11. Characterization of the Human LPIN1-encoded Phosphatidate Phosphatase Isoforms*

    Science.gov (United States)

    Han, Gil-Soo; Carman, George M.

    2010-01-01

    The human LPIN1 gene encodes the protein lipin 1, which possesses phosphatidate (PA) phosphatase (3-sn-phosphatidate phosphohydrolase; EC 3.1.3.4) activity (Han, G.-S., Wu, W.-I., and Carman, G. M. (2006) J. Biol. Chem. 281, 9210–9218). In this work, we characterized human lipin 1 α, β, and γ isoforms that were expressed in Escherichia coli and purified to near homogeneity. PA phosphatase activities of the α, β, and γ isoforms were dependent on Mg2+ or Mn2+ ions at pH 7.5 at 37 °C. The activities were inhibited by concentrations of Mg2+ and Mn2+ above their optimums and by Ca2+, Zn2+, N-ethylmaleimide, propranolol, and the sphingoid bases sphingosine and sphinganine. The activities were thermally labile at temperatures above 40 °C. The α, β, and γ activities followed saturation kinetics with respect to the molar concentration of PA (Km values of 0.35, 0.24, and 0.11 mm, respectively) but followed positive cooperative (Hill number ∼2) kinetics with respect to the surface concentration of PA (Km values of 4.2, 4.5, and 4.3 mol %, respectively) in Triton X-100/PA-mixed micelles. The turnover numbers (kcat) for the α, β, and γ isoforms were 68.8 ± 3.5, 42.8 ± 2.5, and 5.7 ± 0.2 s−1, respectively, whereas their energy of activation values were 14.2, 15.5, and 18.5 kcal/mol, respectively. The isoform activities were dependent on PA as a substrate and required at least one unsaturated fatty acyl moiety for maximum activity. PMID:20231281

  12. Inference of Isoforms from Short Sequence Reads

    Science.gov (United States)

    Feng, Jianxing; Li, Wei; Jiang, Tao

    Due to alternative splicing events in eukaryotic species, the identification of mRNA isoforms (or splicing variants) is a difficult problem. Traditional experimental methods for this purpose are time consuming and cost ineffective. The emerging RNA-Seq technology provides a possible effective method to address this problem. Although the advantages of RNA-Seq over traditional methods in transcriptome analysis have been confirmed by many studies, the inference of isoforms from millions of short sequence reads (e.g., Illumina/Solexa reads) has remained computationally challenging. In this work, we propose a method to calculate the expression levels of isoforms and infer isoforms from short RNA-Seq reads using exon-intron boundary, transcription start site (TSS) and poly-A site (PAS) information. We first formulate the relationship among exons, isoforms, and single-end reads as a convex quadratic program, and then use an efficient algorithm (called IsoInfer) to search for isoforms. IsoInfer can calculate the expression levels of isoforms accurately if all the isoforms are known and infer novel isoforms from scratch. Our experimental tests on known mouse isoforms with both simulated expression levels and reads demonstrate that IsoInfer is able to calculate the expression levels of isoforms with an accuracy comparable to the state-of-the-art statistical method and a 60 times faster speed. Moreover, our tests on both simulated and real reads show that it achieves a good precision and sensitivity in inferring isoforms when given accurate exon-intron boundary, TSS and PAS information, especially for isoforms whose expression levels are significantly high.

  13. The impact of tropomyosins on actin filament assembly is isoform specific.

    Science.gov (United States)

    Janco, Miro; Bonello, Teresa T; Byun, Alex; Coster, Adelle C F; Lebhar, Helene; Dedova, Irina; Gunning, Peter W; Böcking, Till

    2016-07-01

    Tropomyosin (Tpm) is an α helical coiled-coil dimer that forms a co-polymer along the actin filament. Tpm is involved in the regulation of actin's interaction with binding proteins as well as stabilization of the actin filament and its assembly kinetics. Recent studies show that multiple Tpm isoforms also define the functional properties of distinct actin filament populations within a cell. Subtle structural variations within well conserved Tpm isoforms are the key to their functional specificity. Therefore, we purified and characterized a comprehensive set of 8 Tpm isoforms (Tpm1.1, Tpm1.12, Tpm1.6, Tpm1.7, Tpm1.8, Tpm2.1, Tpm3.1, and Tpm4.2), using well-established actin co-sedimentation and pyrene fluorescence polymerization assays. We observed that the apparent affinity (Kd(app)) to filamentous actin varied in all Tpm isoforms between ∼0.1-5 μM with similar values for both, skeletal and cytoskeletal actin filaments. The data did not indicate any correlation between affinity and size of Tpm molecules, however high molecular weight (HMW) isoforms Tpm1.1, Tpm1.6, Tpm1.7 and Tpm2.1, showed ∼3-fold higher cooperativity compared to low molecular weight (LMW) isoforms Tpm1.12, Tpm1.8, Tpm3.1, and Tpm4.2. The rate of actin filament elongation in the presence of Tpm2.1 increased, while all other isoforms decreased the elongation rate by 27-85 %. Our study shows that the biochemical properties of Tpm isoforms are finely tuned and depend on sequence variations in alternatively spliced regions of Tpm molecules.

  14. Kinetic properties of alternatively spliced isoforms of laccase-2 from Tribolium castaneum and Anopheles gambiae.

    Science.gov (United States)

    Gorman, Maureen J; Sullivan, Lucinda I; Nguyen, Thi D T; Dai, Huaien; Arakane, Yasuyuki; Dittmer, Neal T; Syed, Lateef U; Li, Jun; Hua, Duy H; Kanost, Michael R

    2012-03-01

    Laccase-2 is a highly conserved multicopper oxidase that functions in insect cuticle pigmentation and tanning. In many species, alternative splicing gives rise to two laccase-2 isoforms. A comparison of laccase-2 sequences from three orders of insects revealed eleven positions at which there are conserved differences between the A and B isoforms. Homology modeling suggested that these eleven residues are not part of the substrate binding pocket. To determine whether the isoforms have different kinetic properties, we compared the activity of laccase-2 isoforms from Tribolium castaneum and Anopheles gambiae. We partially purified the four laccases as recombinant enzymes and analyzed their ability to oxidize a range of laccase substrates. The predicted endogenous substrates tested were dopamine, N-acetyldopamine (NADA), N-β-alanyldopamine (NBAD) and dopa, which were detected in T. castaneum previously and in A. gambiae as part of this study. Two additional diphenols (catechol and hydroquinone) and one non-phenolic substrate (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)) were also tested. We observed no major differences in substrate specificity between the A and B isoforms. Dopamine, NADA and NBAD were oxidized with catalytic efficiencies ranging from 51 to 550 min⁻¹ mM⁻¹. These results support the hypothesis that dopamine, NADA and NBAD are endogenous substrates for both isoforms of laccase-2. Catalytic efficiencies associated with dopa oxidation were low, ranging from 8 to 30 min⁻¹ mM⁻¹; in comparison, insect tyrosinase oxidized dopa with a catalytic efficiency of 201 min⁻¹ mM⁻¹. We found that dopa had the highest redox potential of the four endogenous substrates, and this property of dopa may explain its poor oxidation by laccase-2. We conclude that laccase-2 splice isoforms are likely to oxidize the same substrates in vivo, and additional experiments will be required to discover any isoform-specific functions.

  15. Quality assurance of GEM foils in the framework of the TPC upgrade in the ALICE experiment

    CERN Document Server

    Ozcelik, Melih Arslan

    2016-01-01

    In the framework of the TPC upgrade of the ALICE Experiment, new readout chambers will be installed during the LHC long shutdown 2, which is scheduled to start in July 2018. The current MWPCs (Multi Wire Proportional Chambers) will be replaced by readout chambers consisting of GEM (Gas Electron Multipliers) foils in order to meet the increasing readout rate requirements. QA (Quality Assurance) tests on the GEMs are performed to classify the foils. In this report we present the work done during the CERN Summer Student Programme 2016.

  16. RCU2-The ALICE TPC readout electronics consolidation for Run2

    CERN Document Server

    Alme, J; Christiansen, P; Yang, S; Lien, J; Velure, A; Rehman, A Ur; Torgersen, C; David, E; Gunji, T; Osterman, L; Ullaland, K; Roed, K; Tarantola, A; Langoy, R; Appelshaeuser, H; Oskarsson, A; Alt, T; Costa, F; Bratrud, L; Zhao, C; Lippmann, C; Torsvik, I Nikolai; Kiss, T

    2013-01-01

    This paper presents the solution for optimization of the ALICE TPC readout for running at full energy in the Run2 period after 2014. For the data taking with heavy ion beams an event readout rate of 400 Hz with a low dead time is envisaged for the ALICE central barrel detectors during these three years. A new component, the Readout Control Unit 2 (RCU2), is being designed to increase the present readout rate by a factor of up to 2.6. The immunity to radiation induced errors will also be significantly improved by the new design.

  17. Silicon ion interactions measured in the E-810 TPC at the AGS

    Energy Technology Data Exchange (ETDEWEB)

    Love, W.A.; Eiseman, S.E.; Etkin, A.; Foley, K.J.; Hackenburg, R.W.; Longacre, R.S.; Morris, T.W.; Platner, E.D.; Saulys, A.C. (Brookhaven National Lab., Upton, NY (USA)); Bonner, B.E.; Buchanan, J.A.; Chiou, C.N.; Clement, J.M.; Corcoran, M.D.; Kruk, J.W.; Mutchler, G.S.; Nessi, M.; Nessi-Tedaldi, F.; Roberts, J.B. (Rice Univ., Houston, TX (USA)); Chan, C.S.; Kramer, M.A.; Zhao, K. (City Univ. of New York, NY (USA)); Hallman, T.J.; Madansky, L. (Johns Hopkins Univ., Baltimore, MD (USA)); Lindenbaum, S.J. (Brookhaven National Lab., Upton, NY (USA) City Univ. of New York, NY (USA)); E-810 Collaboration

    1991-04-01

    The tracking detector of AGS Experiment 810 is a three-piece Time Projection Chamber (TPC) which measures all charged tracks in the forward hemisphere of the nucleon-nucleon center of mass system, i.e. forward of 20 degrees in the lab. A cut at multiplicity 50 was used to select more central collisions, yielding 2291 events from the gold sample, 2170 from the copper. This corresponds to cross sections of 0.59 and 0.20 barns, respectively, defining the 'central' sample for the charged particle distributions presented here. (orig./HSI).

  18. Hydrogen purifier module with membrane support

    Science.gov (United States)

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

    2012-07-24

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

  19. Subunit NDUFV3 is present in two distinct isoforms in mammalian complex I.

    Science.gov (United States)

    Bridges, Hannah R; Mohammed, Khairunnisa; Harbour, Michael E; Hirst, Judy

    2017-03-01

    Complex I (NADH:ubiquinone oxidoreductase) is the first enzyme of the electron transport chain in mammalian mitochondria. Extensive proteomic and structural analyses of complex I from Bos taurus heart mitochondria have shown it comprises 45 subunits encoded on both the nuclear and mitochondrial genomes; 44 of them are different and one is present in two copies. The bovine heart enzyme has provided a model for studying the composition of complex I in other mammalian species, including humans, but the possibility of additional subunits or isoforms in other species or tissues has not been explored. Here, we describe characterization of the complexes I purified from five rat tissues and from a rat hepatoma cell line. We identify a~50kDa isoform of subunit NDUFV3, for which the canonical isoform is only ~10kDa in size. We combine LC-MS and MALDI-TOF mass spectrometry data from two different purification methods (chromatography and immuno-purification) with information from blue native PAGE analyses to show the long isoform is present in the mature complex, but at substoichiometric levels. It is also present in complex I in cultured human cells. We describe evidence that the long isoform is more abundant in both the mitochondria and purified complexes from brain (relative to in heart, liver, kidney and skeletal muscle) and more abundant still in complex I in cultured cells. We propose that the long 50kDa isoform competes with its canonical 10kDa counterpart for a common binding site on the flavoprotein domain of complex I.

  20. Biochemical Characteristics of Three Laccase Isoforms from the Basidiomycete Pleurotus nebrodensis.

    Science.gov (United States)

    Yuan, Xianghe; Tian, Guoting; Zhao, Yongchang; Zhao, Liyan; Wang, Hexiang; Ng, Tzi Bun

    2016-02-06

    The characterization of three laccase isoforms from Pleurotus nebrodensis is described. Isoenzymes Lac1, Lac2 and Lac3 were purified to homogeneity using ion exchange chromatography on DEAE-cellulose, CM-cellulose and Q-Sepharose and a gel filtration step on Superdex 75. The molecular weights of the purified laccases were estimated to be 68, 64 and 51 kDa, respectively. The isoenzymes demonstrated the same optimum pH at 3.0 but slightly different temperature optima: 50-60 °C for Lac1 and Lac3 and 60 °C for Lac2. Lac2 was always more stable than the other two isoforms and exposure to 50 °C for 120 min caused 30% loss in activity. Lac2 was relatively less stable than the other two isoforms when exposed to the pH range of 3.0-8.0 for 24 h, but inactivation only occurred initially, with around 70% residual activity being maintained during the whole process. Oxidative ability towards aromatic compounds varied substantially among the isoforms and each of them displayed preference toward some substrates. Kinetic constants (Km, Kcat) were determined by using a 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assay, with Lac3 showing the best affinity and Lac2 displaying the highest catalytic efficiency. Amino acid sequences from peptides derived from digestion of isoenzymes showed great consistency with laccases in the databases.

  1. Chromatofocusing profile of purified human alpha-fetoprotein and albumin differs from those of crude samples: effect of protein concentration of the elution of the sample.

    Science.gov (United States)

    Leal, J A; Eddy, K B; Keel, B A

    1991-02-01

    Chromatofocusing was utilized to characterize charge microheterogeneity of purified human alpha-fetoprotein (AFP) and human serum albumin (HSA). Crude cord blood samples yielded three isoforms: AFP-IA, IB, and II, with pIs 4.57 (52%), 4.27 (43%), and less than 4.00 (5%), respectively. In contrast, 10 micrograms of purified AFP or 250,000 cpm of 125I-AFP eluted entirely as isoform AFP-II. 125I-AFP focused in the presence of crude cord blood, amniotic fluid, adult male serum, or 25 mg purified HSA resulted in elution profiles similar to those of crude cord blood. Pure AFP focused along with 0.1, 1.0, 5.0, or 10 mg HSA showed a gradual shift from AFP-II to AFP-I. With greater than or equal to 5 mg HSA, isoform I was further resolve into AFP-IA and IB. Similarly, 250,000 cpm of 125I-HSA, which also eluted entirely as isoform II, showed a gradual shift to isoform I when increasing concentrations of unlabeled HSA were added. The resolution of isoform HSA-I in HSA-IA, IB, and IC was again improved with greater than or equal to 5 mg unlabeled HSA. When carrier proteins of varying pI values were chromatofocused along with purified AFP, it was observed that only those proteins with pIs in the range of AFP caused significant alteration in the relative distribution of AFP. We conclude that sample protein concentration and composition must be carefully considered when chromatofocusing is being used for purified samples and when the elution profiles of samples from different origins and varying protein concentrations are being compared.

  2. The FIDIAS project: Development of a Micromegas TPC for the detection of low-energy heavy ions

    Science.gov (United States)

    Iguaz, Francisco José; Panebianco, Stefano; Axiotis, Michael; Druillole, Frédéric; Fanourakis, George; Geralis, Theodoros; Giomataris, Ioannis; Harissopulos, Sotirios; Lagoyannis, Anastasios; Papaevangelou, Thomas

    2014-01-01

    Time Projection Chambers are widely used since many years for tracking and identification of charged particles in high energy physics. We present here a new R&D project, called FIDIAS, meant to investigate the feasibility of a Micromegas TPC for low energy heavy ions detection. In this framework, a TPC prototype based on Micromegas bulk technique has been extensively tested with spontaneous fission source. A deep analysis of the experimental results has been realized leading to a full characterization of the prototype in terms of gain, energy resolution and track reconstruction as a function of three working gas: helium, neon and argon. The encouraging results have also been compared to simulations, showing the Micromegas TPC is a very well suited detector for the detection of heavy ions in nuclear reactions at low energy.

  3. The FIDIAS project: Development of a Micromegas TPC for the detection of low-energy heavy ions

    Energy Technology Data Exchange (ETDEWEB)

    Iguaz, Francisco José [CEA, Centre de Saclay, Institut de Recherche sur les lois Fondamentales de l' Univers, 91191 Gif-sur-Yvette (France); Laboratorio de Fisica Nuclear y Astroparticulas, Universidad de Zaragoza, 50009 Zaragoza (Spain); Panebianco, Stefano, E-mail: stefano.panebianco@cea.fr [CEA, Centre de Saclay, Institut de Recherche sur les lois Fondamentales de l' Univers, 91191 Gif-sur-Yvette (France); Axiotis, Michael [Institute of Nuclear Physics, NCRS Demokritos, 15310 Aghia Paraskevi, Athens (Greece); Druillole, Frédéric [CEA, Centre de Saclay, Institut de Recherche sur les lois Fondamentales de l' Univers, 91191 Gif-sur-Yvette (France); Fanourakis, George; Geralis, Theodoros [Institute of Nuclear Physics, NCRS Demokritos, 15310 Aghia Paraskevi, Athens (Greece); Giomataris, Ioannis [CEA, Centre de Saclay, Institut de Recherche sur les lois Fondamentales de l' Univers, 91191 Gif-sur-Yvette (France); Harissopulos, Sotirios; Lagoyannis, Anastasios [Institute of Nuclear Physics, NCRS Demokritos, 15310 Aghia Paraskevi, Athens (Greece); Papaevangelou, Thomas [CEA, Centre de Saclay, Institut de Recherche sur les lois Fondamentales de l' Univers, 91191 Gif-sur-Yvette (France)

    2014-01-21

    Time Projection Chambers are widely used since many years for tracking and identification of charged particles in high energy physics. We present here a new R and D project, called FIDIAS, meant to investigate the feasibility of a Micromegas TPC for low energy heavy ions detection. In this framework, a TPC prototype based on Micromegas bulk technique has been extensively tested with spontaneous fission source. A deep analysis of the experimental results has been realized leading to a full characterization of the prototype in terms of gain, energy resolution and track reconstruction as a function of three working gas: helium, neon and argon. The encouraging results have also been compared to simulations, showing the Micromegas TPC is a very well suited detector for the detection of heavy ions in nuclear reactions at low energy.

  4. A Time Projection Chamber for High-Rate Experiments: Towards an Upgrade of the ALICE TPC

    CERN Document Server

    Ketzer, Bernhard

    2013-01-01

    A Time Projection Chamber (TPC) is a powerful detector for 3-dimensional tracking and particle identification for ultra-high multiplicity events. It is the central tracking device of many experiments, e.g. the ALICE experiment at CERN. The necessity of a switching electrostatic gate, which prevents ions produced in the amplification region o MWPCs from entering the drift volume, however, restricts its application to trigger rates of the order of 1 kHz. Charge amplification by Gas Electron Multiplier (GEM) foils instead of proportional wires offers an intrinsic suppression of the ion backflow, although not to the same level as a gating grid. Detailed Monte Carlo simulations have shown that the distortions due to residual space charge from back-drifting ions can be limited to a few cm, and thus can be corrected using standard calibration techniques. A prototype GEM-TPC has been built with the largest active volume to date for a detector of this type. It has been commissioned with cosmics and particle beams at t...

  5. Identifying Charged Hadrons on the Relativistic Rise Using the ALICE TPC at LHC

    CERN Document Server

    Gros, Philippe

    2011-01-01

    The chain from hadron collisions to the physics results requires several important links. First the outcome of the collision is measured by the detectors. Then, the signal from the detector is processed and transformed into information relevant for the study of the physics processes. The data is made available to physicists to be analysed and used to improve theories. This thesis presents work done on no most of these steps for the ALICE experiment at LHC. First a study of the main processes in the TPC detector for ALICE was done using simulation and test beam data. The results are shown in paper I. The study was deepened with the analysis of test beam data from a TPC prototype for the ILC, as shown in paper III. Concurrently, a study on the Grid – computing framework for distributed computing and storage resources – was performed. This involved the development of an interface module between the ALICE software AliEn and the ARC software developped in the Nordic countries. This work is presented in paper I...

  6. A TPC-like readout method for high precision muon-tracking using GEM-detectors

    Energy Technology Data Exchange (ETDEWEB)

    Flierl, Bernhard; Biebel, Otmar; Bortfeldt, Jonathan; Hertenberger, Ralf; Klitzner, Felix; Loesel, Philipp; Mueller, Ralph [Ludwig-Maximilians-Universitaet Muenchen (Germany); Zibell, Andre [Julius-Maximilians-Universitaet Wuerzburg (Germany)

    2016-07-01

    Gaseous electron multiplier (GEM) detectors are well suited for tracking of charged particles. Three dimensional tracking in a single layer can be achieved by application of a time-projection-chamber like readout mode (μTPC), if the drift time of the electrons is measured and the position dependence of the arrival time is used to calculate the inclination angle of the track. To optimize the tracking capabilities for ion tracks drift gas mixtures with low drift velocity have been investigated by measuring tracks of cosmic muons in a compact setup of four GEM-detectors of 100 x 100 x 6 mm{sup 3} active volume each and an angular acceptance of -25 to 25 . The setup consists of three detectors with two-dimensional strip readout layers of 0.4 mm pitch and one detector with a single strip readout layer of 0.25 mm pitch. All strips are readout by APV25 frontend boards and the amplification stage in the detectors consists of three GEM-foils. Tracks are reconstructed by the μTPC-method in one of the detectors and are then compared to the prediction from the other three detectors defined by the center of charge in every detector. We report our study of Argon and Helium based noble gas mixtures with carbon-dioxide as quencher.

  7. NEXT: Searching for the neutrinoless double beta decay with a gas-xenon TPC

    Energy Technology Data Exchange (ETDEWEB)

    Novella, P, E-mail: pau.novella@ciemat.e [CIEMAT, Av. Complutense 22, 28040 Madrid (Spain)

    2010-01-01

    Although different techniques are used to search for the neutrinoless double beta decay, the common challenges for all the existing or planned experiments are to achieve a good energy resolution and large background rejection factors. The NEXT collaboration addresses these two challenges with a high-pressure gas-Xenon TPC. Natural Xenon consists of almost 9% of {sup 136}Xe, a {beta}{beta}{sup 0{nu}} candidate emitter, and can be easily enriched. When used as a calorimeter, {sup 136}Xe yields an excellent energy resolution. This fact, combined with the expected long life of the {beta}{beta}{sup 2{nu}} mode, accounts for negligible intrinsic backgrounds up to masses of 1 ton. Furthermore, external backgrounds can be rejected with high efficiency by means of the electron tracking capabilities of the TPC. A detector containing about 100 kg of enriched Xenon is expected to be installed at Canfranc Underground Laboratory (LSC) within the next 5 years, with the twofold aim of exploring the degenerated hierarchy of the neutrino mass and providing deep understanding of the experimental techniques which allow extrapolation to larger detectors.

  8. Gas Gain Measurements from a Negative Ion TPC X-ray Polarimeter

    CERN Document Server

    Prieskorn, Z; Kaaret, P E; Black, J K; Jahoda, K

    2011-01-01

    Gas-based time projection chambers (TPCs) have been shown to be highly sensitive X-ray polarimeters having excellent quantum efficiency while at the same time achieving large modulation factors. To observe polarization of the prompt X-ray emission of a Gamma-ray burst (GRB), a large area detector is needed. Diffusion of the electron cloud in a standard TPC could be prohibitive to measuring good modulation when the drift distance is large. Therefore, we propose using a negative ion TPC (NITPC) with Nitromethane (CH3NO2) as the electron capture agent. The diffusion of negative ions is reduced over that of electrons due to the thermal coupling of the negative ions to the surrounding gas. This allows for larger area detectors as the drift distance can be increased without degrading polarimeter modulation. Negative ions also travel ~200 times slower than electrons, allowing the readout electronics to operate slower, resulting in a reduction of instrument power. To optimize the NITPC design, we have measured gas ga...

  9. Isolation and characterization of patatin isoforms

    NARCIS (Netherlands)

    Pots, A.M.; Gruppen, H.; Hessing, M.; Boekel, M.A.J.S. van; Voragen, A.G.J.

    1999-01-01

    Patatin has, so far, been considered a homogeneous group of proteins. A comparison of the isoforms in terms of structural properties or stability has not been reported. A method to obtain various isoform fractions as well as a comparison of the physicochemical properties of these pools is presented.

  10. Ceramic materials purified by experimental method

    Science.gov (United States)

    1965-01-01

    Crystalline ceramic materials are purified for use as high-temperature electrical insulators. Any impurities migrate to the cathode when a dc voltage is applied across the material while it is heated in an inert gas atmosphere.

  11. Building a large-area GEM-based readout chamber for the upgrade of the ALICE TPC

    CERN Document Server

    Gasik, Piotr

    2016-01-01

    A large Time Projection Chamber (TPC) is the main device for tracking and charged-particle identification in the ALICE experiment at the CERN LHC. After the second long shutdown in 2019-2020, the LHC will deliver Pb beams colliding at an interaction rate up to 50 kHz, which is about a factor of 100 above the present read-out rate of the TPC. To fully exploit the LHC potential the TPC will be upgraded based on the Gas Electron Multiplier (GEM) technology. A prototype of an ALICE TPC Outer Read-Out Chamber (OROC) was equipped with twelve large-size GEM foils as amplification stage to demonstrate the feasibility of replacing the current Multi Wire Proportional Chambers with the new technology. With a total area of $\\sim$0.76 m$^2$ it is the largest GEM-based detector built to date. The GEM OROC was installed within a test field cage and commissioned with radioactive sources.

  12. Building a large-area GEM-based readout chamber for the upgrade of the ALICE TPC

    Science.gov (United States)

    Gasik, P.

    2017-02-01

    A large Time Projection Chamber (TPC) is the main device for tracking and charged-particle identification in the ALICE experiment at the CERN LHC. After the second long shutdown in 2019-2020, the LHC will deliver Pb beams colliding at an interaction rate up to 50 kHz, which is about a factor of 100 above the present read-out rate of the TPC. To fully exploit the LHC potential the TPC will be upgraded based on the Gas Electron Multiplier (GEM) technology. A prototype of an ALICE TPC Outer Read-Out Chamber (OROC) was equipped with twelve large-size GEM foils as amplification stage to demonstrate the feasibility of replacing the current Multi Wire Proportional Chambers with the new technology. With a total area of ∼0.76 m2 it is the largest GEM-based detector built to date. The GEM OROC was installed within a test field cage and commissioned with radioactive sources.

  13. Probing the fourth neutrino existence by neutral current oscillometry in the spherical gaseous TPC

    CERN Document Server

    Vergados, J D; Novikov, Yu N

    2011-01-01

    It is shown that, if the "new neutrino" implied by the Reactor Neutrino Anomaly exists and is in fact characterized by the suggested relatively high mass squared difference and reasonably large mixing angle, it should clearly reveal itself in the oscillometry measurements. For a judicious neutrino source the "new oscillation length L14 is expected shorter than 3m. Thus the needed measurements can be implemented with a gaseous spherical TPC of modest dimensions with a very good energy and position resolution, detecting nuclear recoils following the coherent neutrino-nucleus elastic scattering. The best candidates for oscillometry, yielding both monochromatic neutrinos as well as antineutrinos, are discussed. A sensitivity in the mixing angle theta14, (sin(2\\theta14))^2=0.1 (99 %), can be reached after a few months of data handling.

  14. Neutral Current Coherent Cross Sections -- Implications on Gaseous Spherical TPC's for detecting SN and Earth neutrinos

    CERN Document Server

    Giomataris, Y

    2016-01-01

    The detection of galactic supernova (SN) neutrinos represents one of the future frontiers of low-energy neutrino physics and astrophysics. The neutron coherence of neutral currents (NC) allows quite large cross sections in the case of neutron rich targets, which can be exploited in detecting earth and sky neutrinos by measuring nuclear recoils. They are relatively cheap and easy to maintain. The relevant NC cross sections are not dependent on flavor conversions and, thus, their measurement will provide useful information about the neutrino source. In particular they will yield information about the primary neutrino fluxes and perhaps about the spectrum after flavor conversions in neutrino sphere.They might also provide some clues about the neutrino mass hierarchy. The advantages of large gaseous low threshold and high resolution detectors with time projection counters (TPC) are discussed.

  15. The $\\mu$TPC Method: Improving the Position Resolution of Neutron Detectors Based on MPGDs

    CERN Document Server

    Pfeiffer, Dorothea; Birch, Jens; Hall-Wilton, Richard; Höglund, Carina; Hultman, Lars; Iakovidis, George; Oliveri, Eraldo; Oksanen, Esko; Ropelewski, Leszek; Thuiner, Patrik

    2015-01-01

    Due to the Helium-3 crisis, alternatives to the standard neutron detection techniques are becoming urgent. In addition, the instruments of the European Spallation Source (ESS) require advances in the state of the art of neutron detection. The instruments need detectors with excellent neutron detection efficiency, high-rate capabilities and unprecedented spatial resolution. The Macromolecular Crystallography instrument (NMX) requires a position resolution in the order of 200 um over a wide angular range of incoming neutrons. Solid converters in combination with Micro Pattern Gaseous Detectors (MPGDs) are proposed to meet the new requirements. Charged particles rising from the neutron capture have usually ranges larger than several millimetres in gas. This is apparently in contrast with the requirements for the position resolution. In this paper, we present an analysis technique, new in the field of neutron detection, based on the Time Projection Chamber (TPC) concept. Using a standard Single-GEM with the catho...

  16. MIMAC-He3 : MIcro-tpc MAtrix of Chambers of He3

    CERN Document Server

    Santos, D; Lamy, T; Mayet, F; Moulin, E

    2016-01-01

    The project of a micro-TPC matrix of chambers of He3 for direct detection of non-baryonic dark matter is outlined. The privileged properties of He3 are highlighted. The double detection (ionization - projection of tracks) will assure the electron-recoil discrimination. The complementarity of MIMAC-He3 for supersymmetric dark matter search with respect to other experiments is illustrated. The modular character of the detector allows to have different gases to get A-dependence. The pressure degreee of freedom gives the possibility to work at high and low pressure. The low pressure regime gives the possibility to get the directionality of the tracks. The first measurements of ionization at very few keVs for He3 in He4 gas are described.

  17. Improved TPB-coated Light Guides for Liquid Argon TPC Light Detection Systems

    CERN Document Server

    Moss, Z; Collin, G; Conrad, J M; Jones, B J P; Moon, J; Toups, M; Wongjirad, T

    2014-01-01

    Scintillation light produced in liquid argon (LAr) must be shifted from 128 nm to visible wavelengths in light detection systems used for liquid argon time-projection chambers (LArTPCs). To date, LArTPC light collection systems have employed tetra phenyl butadiene (TPB) coatings on photomultiplier tubes (PMTs) or plates placed in front of the PMTs. Recently, a new approach using TPB-coated light guides was proposed. In this paper, we report on light guides with improved attenuation lengths above 100 cm when measured in air. This is an important step in the development of meter-scale light guides for future LArTPCs. Improvements come from using a new acrylic-based coating, diamond-polished cast UV transmitting acrylic bars, and a hand-dipping technique to coat the bars.

  18. Development and Characterization of a Multi-APD Xenon Electroluminescence TPC

    CERN Document Server

    Lux, T; Ballester, O; Bordoni, S; Gil-Botella, I; Hamer, N; Illa, J; Mañas, G Jover; Martin-Mari, C; Palomares, C; Rico, J; Sanchez, F; Santorelli, R; Verdugo, A

    2014-01-01

    The performance of an electroluminescence (EL) time projection chamber (TPC) with a multi avalanche photodiode (APD) readout was studied in pure xenon at 3.8 bar. Intercalibration and reconstruction methods were developed and applied to the data yielding energy resolutions as good as 5.3$+-$0.1 % FWHM for 59.5 keV gammas from 241-Am. The result was verified with a Monte Carlo (MC) based on Geant4 and Penelope predicting 5.2 % FWHM for the used setup. Point resolutions of about 0.5 mm were obtained by a pitch of 15 mm between the APDs. The results show that a multi-APD readout is a competitive technology for EL detectors filled with pure xenon with possible applications as Compton Cameras.

  19. MIMAC : A micro-tpc matrix for directional detection of dark matter

    CERN Document Server

    Santos, D; Bosson, G; Bouly, J L; Bourrion, O; Fourel, Ch; Grignon, C; Guillaudin, O; Mayet, F; Richer, J P; Delbart, A; Ferrer, E; Giomataris, I; Iguaz, F J; Mols, J P; Golabek, C; Lebreton, L

    2011-01-01

    Directional detection of non-baryonic Dark Matter is a promising search strategy for discriminating WIMP events from background. However, this strategy requires both a precise measurement of the energy down to a few keV and 3D reconstruction of tracks down to a few mm. To achieve this goal, the MIMAC project has been developed. It is based on a gaseous micro-TPC matrix, filled with CF4 and CHF3. The first results on low energy nuclear recoils (H, F) obtained with a low mono-energetic neutron field are presented. The discovery potential of this search strategy is discussed and illustrated by a realistic case accessible to MIMAC.

  20. MIMAC: A micro-tpc matrix for directional detection of dark matter

    Energy Technology Data Exchange (ETDEWEB)

    Santos, D; Billard, J; Bosson, G; Bouly, J L; Bourrion, O; Fourel, Ch; Grignon, C; Guillaudin, O; Mayet, F; Richer, J P [LPSC, Universite Joseph Fourier Grenoble 1, CNRS/IN2P3, Institut Polytechnique de Grenoble (France); Delbart, A; Ferrer, E; Giomataris, I; Iguaz, F J; Mols, J P [IRFU,CEA Saclay, 91191 Gif-sur-Yvette cedex (France); Golabek, C; Lebreton, L, E-mail: Daniel.Santos@lpsc.in2p3.fr [LMDN, IRSN Cadarache, 13115 Saint-Paul-Lez-Durance (France)

    2011-08-10

    Directional detection of non-baryonic Dark Matter is a promising search strategy for discriminating WIMP events from background. However, this strategy requires both a precise measurement of the energy down to a few keV and 3D reconstruction of tracks down to a few mm. To achieve this goal, the MIMAC project has been developed. It is based on a gaseous micro-TPC matrix, filled with CF{sub 4} and CHF{sub 3}. The first results on low energy nuclear recoils ({sup 1}H and {sup 19}F) obtained with a low mono-energetic neutron field are presented. The discovery potential of this search strategy is discussed and illustrated by a realistic case accessible to MIMAC.

  1. MIMAC: A micro-tpc matrix project for directional detection of dark matter

    CERN Document Server

    Santos, D; Bosson, G; Bouly, J L; Bourrion, O; Fourel, Ch; Guillaudin, O; Mayet, F; Richer, J P; Delbart, A; Ferrer, E; Giomataris, I; Iguaz, F J; Mols, J P; Golabek, C; Lebreton, L

    2011-01-01

    Directional detection of non-baryonic DarkMatter is a promising search strategy for discriminating WIMP events from background ones. This strategy requires both a measurement of the recoil energy down to a few keV and 3D reconstruction of tracks down to a few mm. The MIMAC project, based on a micro-TPC matrix, filled with CF4 and CHF3 is being developed. The first results of a chamber prototype of this matrix, on low energy nuclear recoils (1H and 19F) obtained with mono-energetic neutron fields are presented. The discovery potential of this search strategy is illustrated by a realistic case accessible to MIMAC.

  2. MIMAC: MIcro-tpc MAtrix of Chambers for dark matter directional detection

    CERN Document Server

    Santos, D; Bouly, J L; Bourrion, O; Fourel, Ch; Guillaudin, O; Lamblin, J; Mayet, F; Muraz, J F; Richer, J P; Riffard, Q; Lebreton, L; Maire, D; Busto, J; Brunner, J; Fouchez, D

    2013-01-01

    Directional detection of non-baryonic Dark Matter is a promising search strategy for discriminating WIMP events from neutrons, the ultimate background for dark matter direct detection. This strategy requires both a precise measurement of the energy down to a few keV and 3D reconstruction of tracks down to a few mm. The MIMAC (MIcro-tpc MAtrix of Chambers) collaboration has developed in the last years an original prototype detector based on the direct coupling of large pixelized micromegas with a special developed fast self-triggered electronics showing the feasibility of a new generation of directional detectors. The first bi-chamber prototype has been installed at Modane, underground laboratory in June 2012. The first undergournd background events, the gain stability and calibration are shown. The first spectrum of nuclear recoils showing 3D tracks coming from the radon progeny is presented.

  3. MIMAC: A micro-tpc matrix for dark matter directional detection

    CERN Document Server

    Santos, D; Bosson, G; Bouly, J L; Bourrion, O; Fourel, C; Guillaudin, O; Lamblin, J; Muraz, J F; Mayet, F; Richer, J P; Riffard, Q; Ferrer, E; Giomataris, I; Iguaz, F J; Lebreton, L; Maire, D

    2013-01-01

    The dark matter directional detection opens a new field in cosmology bringing the possibility to build a map of nuclear recoils that would be able to explore the galactic dark matter halo giving access to a particle characterization of such matter and the shape of the halo. The MIMAC (MIcro-tpc MAtrix of Chambers) collaboration has developed in the last years an original prototype detector based on the direct coupling of large pixelized micromegas with a devoted fast self-triggered electronics showing the feasibility of a new generation of directional detectors. The discovery potential of this search strategy is discussed and illustrated. In June 2012, the first bi-chamber prototype has been installed at Modane Underground Laboratory (LSM) and the first underground background events, the gain stability and calibration are shown.

  4. MIMAC : A micro-tpc matrix for directional detection of dark matter

    CERN Document Server

    Santos, D; Bosson, G; Bourrion, O; Grignon, C; Guillaudin, O; Mayet, F; Richer, J P; Ferrer, E; Giomataris, I; Iguaz, F J; Mols, J P; Allaoua, A; Golabek, C; Lebreton, L

    2010-01-01

    Directional detection of non-baryonic Dark Matter is a promising search strategy for discriminating WIMP events from background. However, this strategy requires both a precise measurement of the energy down to a few keV and 3D reconstruction of tracks down to a few mm. To achieve this goal, the MIMAC project has been developed. It is based on a gaseous micro-TPC matrix, filled with 3He, CF4 and/or C4H10. The first results on low energy nuclear recoils (1H and 19F) obtained with a low mono-energetic neutron field are presented. The discovery potential of this search strategy is discussed and illustrated by a realistic case accessible to MIMAC.

  5. Amplification and Scintillation Properties of Oxygen-Rich Gas Mixtures for Optical-TPC Applications

    CERN Document Server

    Weissman, L; Chechik, R; Dangendorf, V; Gai, M; Tittelmeier, K; Weller, H R

    2006-01-01

    We studied electron amplification and light emission from avalanches in oxygen-containing gas mixtures. The mixtures investigated in this work included, among others, CO2 and N2O mixed with Triethylamine (TEA) or N2. Double-Step Parallel Gap (DSPG) multipliers and THick Gas Electron Multipliers (THGEM) were investigated. High light yields were measured from CO2+N2 and CO2+TEA, though with different emission spectra. We observed the characteristic wave-length emission of N2 and of TEA and used a polymer wave-length shifter to convert TEA UV-light into the visible spectrum. The results of these measurements indicate the applicability of optical recording of ionizing tracks in a TPC target-detector designed to study the cross section of the 16O(g,a)12C reaction, a central problem in nuclear astrophysics.

  6. Directional detection of Dark Matter with the MIcro-tpc MAtrix of Chambers

    CERN Document Server

    Couturier, C; Naraghi, F; Riffard, Q; Santos, D; Sauzet, N; Colas, P; Ribas, E Ferrer; Giomataris, I; Busto, J; Fouchez, D; Tao, C; Zhou, N

    2016-01-01

    Particles weakly interacting with ordinary matter, with an associated mass of the order of an atomic nucleus (WIMPs), are plausible candidates for Dark Matter. The direct detection of an elastic collision of a target nuclei induced by one of these WIMPs has to be discriminated from the signal produced by the neutrons, which leaves the same signal in a detector. The MIMAC (MIcro-tpc MAtrix of Chambers) collaboration has developed an original prototype detector which combines a large pixelated Micromegas coupled with a fast, self-triggering, electronics. Aspects of the two-chamber module in operation in the Modane Underground Laboratory are presented: calibration, characterization of the $^{222}$Rn progeny. A new test bench combining a MIMAC chamber with the COMIMAC portable quenching line has been set up to characterize the 3D tracks of low energy ions in the MIMAC gas mixture: the preliminary results thereof are presented. Future steps are briefly discussed.

  7. TPC-like readout for thermal neutron detection using a GEM-detector

    CERN Document Server

    Flierl, Bernhard; Hertenberger, Ralf; Zeitelhack, Karl

    2015-01-01

    Spatial resolution of less than 200 um is challenging for thermal neutron detection. A novel readout scheme based on the time-projection-chamber (TPC) concept is used in a gaseous electron multiplier (GEM) detector. Thermal neutrons are captured in a single 2 um thick Boron-10 converter cathode and secondary Helium and Lithium ions are produced with a combined energy of 2.8 MeV. These ions have sufficient energy to form straight tracks of several mm length. With a time resolving 2-dimensional readout of 400 um pitch in both directions, based on APV25 chips, the ions are tracked and their respective origin in the cathode converter foil is reconstructed. Using an Ar-CO2 93:7% gas mixture, a resolution of 100 um (FWHM 235 um) has been observed with a triple GEM-detector setup at the Garching neutron source (FRMII) for neutrons of 4.7 Angstrom.

  8. SAMPA Chip: the New 32 Channels ASIC for the ALICE TPC and MCH Upgrades

    Science.gov (United States)

    Adolfsson, J.; Ayala Pabon, A.; Bregant, M.; Britton, C.; Brulin, G.; Carvalho, D.; Chambert, V.; Chinellato, D.; Espagnon, B.; Hernandez Herrera, H. D.; Ljubicic, T.; Mahmood, S. M.; Mjörnmark, U.; Moraes, D.; Munhoz, M. G.; Noël, G.; Oskarsson, A.; Osterman, L.; Pilyar, A.; Read, K.; Ruette, A.; Russo, P.; Sanches, B. C. S.; Severo, L.; Silvermyr, D.; Suire, C.; Tambave, G. J.; Tun-Lanoë, K. M. M.; van Noije, W.; Velure, A.; Vereschagin, S.; Wanlin, E.; Weber, T. O.; Zaporozhets, S.

    2017-04-01

    This paper presents the test results of the second prototype of SAMPA, the ASIC designed for the upgrade of read-out front end electronics of the ALICE Time Projection Chamber (TPC) and Muon Chamber (MCH). SAMPA is made in a 130 nm CMOS technology with 1.25 V nominal voltage supply and provides 32 channels, with selectable input polarity, and three possible combinations of shaping time and sensitivity. Each channel consists of a Charge Sensitive Amplifier, a semi-Gaussian shaper and a 10-bit ADC; a Digital Signal Processor provides digital filtering and compression capability. In the second prototype run both full chip and single test blocks were fabricated, allowing block characterization and full system behaviour studies. Experimental results are here presented showing agreement with requirements for both the blocks and the full chip.

  9. WA105: A large demonstrator of a liquid argon dual phase TPC

    Science.gov (United States)

    Zambelli, L.; Murphy, S.; WA105 Collaboration

    2017-09-01

    The Liquid argon technology has been chosen for the DUNE underground experiment for the study of neutrino oscillations, neutrino astrophysics and proton decay. This detector has excellent tracking and calorimetric capabilities much superior to currently operating neutrino detectors. WA105 is a large demonstrator of the dual-phase liquid argon TPC based on the GLACIER design, with a 6×6×6 m3 (appr. 300t) active volume. Its construction and operation test scalable solutions for the crucial aspects of this detector: ultra-high argon purity in non-evacuable tanks, long drifts, very high drift voltages, large area MPGD, cold preamplifiers. The TPC will be built inside a tank based on industrial LNG technology. Electrons produced in the liquid argon are extracted in the gas phase. Here, a readout plane based on Large Electron Multipliers (LEM’s) provides amplification before the charge collection onto an anode plane with strip readout. This highly cost effective solution provides excellent imaging capabilities with equal charge sharing on both views. PMTs located at the bottom of the tank containing the liquid argon provide the readout of the scintillation light. This demonstrator is an industrial prototype of the design proposed for a large underground detector. WA105 is under construction at CERN and will be exposed to a charged particle beam (0.5 - 20 GeV/c) in the North Area in 2018. The data will provide necessary calibration of the detector performances and benchmark sophisticated reconstruction algorithms. This project is a crucial milestone for the long baseline neutrino program DUNE.

  10. GEM-based TPC with CCD imaging for directional dark matter detection

    Science.gov (United States)

    Phan, N. S.; Lauer, R. J.; Lee, E. R.; Loomba, D.; Matthews, J. A. J.; Miller, E. H.

    2016-11-01

    The most mature directional dark matter experiments at present all utilize low-pressure gas Time Projection Chamber (TPC) technologies. We discuss some of the challenges for this technology, for which balancing the goal of achieving the best sensitivity with that of cost effective scale-up requires optimization over a large parameter space. Critical for this are the precision measurements of the fundamental properties of both electron and nuclear recoil tracks down to the lowest detectable energies. Such measurements are necessary to provide a benchmark for background discrimination and directional sensitivity that could be used for future optimization studies for directional dark matter experiments. In this paper we describe a small, high resolution, high signal-to-noise GEM-based TPC with a 2D CCD readout designed for this goal. The performance of the detector was characterized using alpha particles, X-rays, gamma-rays, and neutrons, enabling detailed measurements of electron and nuclear recoil tracks. Stable effective gas gains of greater than 1 × 105 were obtained in 100 Torr of pure CF4 by a cascade of three standard CERN GEMs each with a 140 μm pitch. The high signal-to-noise and sub-millimeter spatial resolution of the GEM amplification and CCD readout, together with low diffusion, allow for excellent background discrimination between electron and nuclear recoils down below ∼10 keVee (∼23 keVr fluorine recoil). Even lower thresholds, necessary for the detection of low mass WIMPs for example, might be achieved by lowering the pressure and utilizing full 3D track reconstruction. These and other paths for improvements are discussed, as are possible fundamental limitations imposed by the physics of energy loss.

  11. Prokaryotic expression and characterization of a pea actin isoform (PEAcl) fused to GFP

    Institute of Scientific and Technical Information of China (English)

    ZHANG Shaobin; REN Dongtao; XU Xiaojing; LIU Guoqin

    2004-01-01

    Actins widely exist in eukaryotic cells and play important roles in many living activities. As there are many kinds of actin isoforms in plant cells, it is difficult to purify each actin isoform in sufficient quantities for analyzing its physicochemical properties. In the present study, a pea(Pisum Sativum L.) actin isoform (PEAc1) fused to His-tag at its amino terminus and GFP (green fluorescent protein) at its Carboxyl terminus were expressed in E. Coli in inclusion bodies. The fusion protein (PEAc1-GFP) was highly purified with the yield of above 2 mg/L culture by dissolving inclusions in 8 mol/L urea, renaturing by dialysis in a gradient of urea, and affinity binding to Ni-resin. The purified mono meric PEAc1-GFP could efficiently bind on Dnase Ⅰ and inhibit the latter's enzyme activity. PEAc1-GFP could polymerize into green fluorescent filamentous structures (F-PEAc1-GFP), which could be labeled by TRITC-phalloidin, a specific agent for observing microfilaments. The PEAc1-GFP polymerization curve was identical with that of chicken skeletal muscle actin. The critical concentration for PEAc1-GFP to polymerize into filaments is 0.24 μmol/L. The F-PEAc1-GFP could stimulate myosin Mg-ATPase activity in a protein concentration dependant manner (about 4 folds at1 mg/mL F-PEAc1-GFP). The results above show that the PEAcl fused to GFP retained the assembly characteristic of actin, indicating that gene fusion, prokaryotic expression,denaturation and renaturation, and affinity chromatography is a useful strategy for obtaining plant actin isoform proteins in a large amount.

  12. Preparation and Affinity-Purification of Supervillin Isoform 4 (SV4) Specific Polyclonal Antibodies.

    Science.gov (United States)

    Chen, Xueran; Li, Hao; Wang, Hongzhi; Yang, Haoran; Ye, Fang; Liang, Chaozhao; Fang, Zhiyou

    2016-04-01

    Human Supervillin isoform 4 (SV4), a bigger splicing isoform of Supervillin, contains extra coding exons 3, 4 and 5 (E345), compared to Supervillin isoform 1. Although previous studies have shown that SV4 associated with membrane and cytoskeleton, regulated cell migration and cell survival, its functions are still largely unknown. To broaden our understanding, SV4 specific antibody is important for further study in signaling pathway. The His-SV4 (E345) and GST-SV4 (E345) fusion proteins, which contained SV4 specific domain E345, were purified from bacteria. The His-SV4 (E345) proteins were injected in rabbits as immunogen to produce anti-SV4 serum, and SV4 antibodies were purified by GST-SV4 (E345) proteins cross-linked to affinity resins. SV4 antibodies exclusively recognized SV4 protein both in vitro and in vivo through multi-step testing by ELISA, western blot, immunoprecipitation, and immunofluorescence. Taken together, our data demonstrate a novel SV4-specific polyclonal antibody which will provide a useful tool for further characterization of SV4 function.

  13. FSH isoform pattern in classic galactosemia

    OpenAIRE

    Gubbels, C.S.; Thomas, C. M.; Wodzig, K.W.H.; Olthaar, A. J.; Jaeken, J.; Sweep, F.C.; Rubio-Gozalbo, M.E.

    2010-01-01

    Female classic galactosemia patients suffer from primary ovarian insufficiency (POI). The cause for this long-term complication is not fully understood. One of the proposed mechanisms is that hypoglycosylation of complex molecules, a known secondary phenomenon of galactosemia, leads to FSH dysfunction. An earlier study showed less acidic isoforms of FSH in serum samples of two classic galactosemia patients compared to controls, indicating hypoglycosylation. In this study, FSH isoform patterns...

  14. Sample displacement chromatography of plasmid DNA isoforms.

    Science.gov (United States)

    Černigoj, Urh; Martinuč, Urška; Cardoso, Sara; Sekirnik, Rok; Krajnc, Nika Lendero; Štrancar, Aleš

    2015-10-02

    Sample displacement chromatography (SDC) is a chromatographic technique that utilises different relative binding affinities of components in a sample mixture and has been widely studied in the context of peptide and protein purification. Here, we report a use of SDC to separate plasmid DNA (pDNA) isoforms under overloading conditions, where supercoiled (sc) isoform acts as a displacer of open circular (oc) or linear isoform. Since displacement is more efficient when mass transfer between stationary and mobile chromatographic phases is not limited by diffusion, we investigated convective interaction media (CIM) monoliths as stationary phases for pDNA isoform separation. CIM monoliths with different hydrophobicities and thus different binding affinities for pDNA (CIM C4 HLD, CIM-histamine and CIM-pyridine) were tested under hydrophobic interaction chromatography (HIC) conditions. SD efficiency for pDNA isoform separation was shown to be dependent on column selectivity for individual isoform, column efficiency and on ammonium sulfate (AS) concentration in loading buffer (binding strength). SD and negative mode elution often operate in parallel, therefore negative mode elution additionally influences the efficiency of the overall purification process. Optimisation of chromatographic conditions achieved 98% sc pDNA homogeneity and a dynamic binding capacity of over 1mg/mL at a relatively low concentration of AS. SDC was successfully implemented for the enrichment of sc pDNA for plasmid vectors of different sizes, and for separation of linear and and sc isoforms, independently of oc:sc isoform ratio, and flow-rate used. This study therefore identifies SDC as a promising new approach to large-scale pDNA purification, which is compatible with continuous, multicolumn chromatography systems, and could therefore be used to increase productivity of pDNA production in the future. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Fast TPC Online Tracking on GPUs and Asynchronous Data Processing in the ALICE HLT to facilitate Online Calibration

    Science.gov (United States)

    Rohr, David; Gorbunov, Sergey; Krzewicki, Mikolaj; Breitner, Timo; Kretz, Matthias; Lindenstruth, Volker

    2015-12-01

    ALICE (A Large Heavy Ion Experiment) is one of the four major experiments at the Large Hadron Collider (LHC) at CERN, which is today the most powerful particle accelerator worldwide. The High Level Trigger (HLT) is an online compute farm of about 200 nodes, which reconstructs events measured by the ALICE detector in real-time. The HLT uses a custom online data-transport framework to distribute data and workload among the compute nodes. ALICE employs several calibration-sensitive subdetectors, e.g. the TPC (Time Projection Chamber). For a precise reconstruction, the HLT has to perform the calibration online. Online- calibration can make certain Offline calibration steps obsolete and can thus speed up Offline analysis. Looking forward to ALICE Run III starting in 2020, online calibration becomes a necessity. The main detector used for track reconstruction is the TPC. Reconstructing the trajectories in the TPC is the most compute-intense step during event reconstruction. Therefore, a fast tracking implementation is of great importance. Reconstructed TPC tracks build the basis for the calibration making a fast online-tracking mandatory. We present several components developed for the ALICE High Level Trigger to perform fast event reconstruction and to provide features required for online calibration. As first topic, we present our TPC tracker, which employs GPUs to speed up the processing, and which bases on a Cellular Automaton and on the Kalman filter. Our TPC tracking algorithm has been successfully used in 2011 and 2012 in the lead-lead and the proton-lead runs. We have improved it to leverage features of newer GPUs and we have ported it to support OpenCL, CUDA, and CPUs with a single common source code. This makes us vendor independent. As second topic, we present framework extensions required for online calibration. The extensions, however, are generic and can be used for other purposes as well. We have extended the framework to support asynchronous compute

  16. Methods and results for calibration and track separation of a GEM based TPC using an UV-laser

    Energy Technology Data Exchange (ETDEWEB)

    Ball, Markus

    2008-12-15

    In the last 30 years high energy physics could write an impressive story of success. Since the introduction of the Standard Model (SM), it has met every experimental test. However the final confirmation has to prove the mechanism of electroweak symmetry breaking, which could not be confirmed yet. The most favored theory, which includes the introduction of a Higgs field, could not be verified experimentally. Furthermore there is clear evidence, that the SM is only a low energy description of nature and its principles, as the SM describes only 4 % of the known matter in the universe. There are two different approaches in accelerator driven high energy physics to clarify the open questions. The Large Hadron Collider (LHC) have a good opportunity to measure some of the missing pieces with its high center of mass energy. The International Linear Collider (ILC) will then measure their parameters with high precision. To guarantee this high precision the detectors have to be able to identify every single particle and determine its properties with high accuracy. These high requirements to the single detectors as well as the interconnectivity between all detectors are summarised by the concept of particle flow (PFLOW). This means that all particles must be separable, which includes in particular the main tracking device. A possible candidate for the central tracking device is a Time Projection Chamber (TPC). In this work a TPC with Gas Electron Multipliers (GEM) as gas amplification system was used. The GEMs replace the conventional wire amplification system of the TPC. In this PhD work a method to determine the drift velocity of a TPC was developed and tested using an ultraviolet laser. To ensure a high accuracy of the method all relevant gas parameters were measured with a slow control system. Furthermore the laser was used to investigate the separation capability of nearby tracks. Therefore an existing TPC prototype, which was developed to operate in a 5 T magnet facility

  17. Screening of lectins from South American plants used as affinity ligands to purify rhEPO

    Directory of Open Access Journals (Sweden)

    G.I. Amadeo

    2003-03-01

    Full Text Available Two groups of isoforms of rhEPO, at a concentration of 300 µg/ml, were tested as putative inhibitors of the lectinic hemagglutination reaction in order to obtain affinity ligand(s for hormone purification: groups I (pI: 3.80; 3.89; 3.95; 4.07, 4.15 and 4.26 and groups II (pI: 4.15, 4.26; 4.38; 4.51; 4.72 and 4.93 Crude extracts from the vegetable materials Abrus precatorious (Abrin, Artocarpus incisa (Frutalin, Artocarpus integrifolia (Jacalin, Canavalia ensiformes (ConA, Canavalia brasiliensis (Conbr, Cratylia floribunda, Dioclea altissima (DAL, Dioclea grandiflora (DGL, Erythrina vellutina (EVL, Erythrina cristagalli, Lutaelburgia auriculata (lectin not fully characterized yet, Lycopersicum esculentum (LEA, Phaseolus vulgaris (PHA, Ricinus communis (Ricin and Triticum vulgaris (WGA were used. Only some of the galactose-specific lectins and the GlcNAc-specific lectins showed rapid full inhibition of the hemagglutination reaction for the less acidic isoforms and the total isoforms of rhEPO, respectively. On this basis, the selected lectins were purified by affinity chromatoghraphy and covalently coupled to cyanogen bromide activated Sepharose® (Amersham-Pharmacia. CHO.K1 cell culture supernatant containing rhEPO was loaded onto the lectin resins and the recoveries were calculated by using specific elutions.

  18. Bacteria that purify sludge; Des bacteries epuratrices

    Energy Technology Data Exchange (ETDEWEB)

    Peignen-Seraline, P.; Manem, J. [Cirsee, Lyonnaise des Eaux, 92 - Nanterre (France)

    1997-03-01

    Inherent in water purification processes, the formation of sludges is intensively studied. Recently, original bacteria have been observed by searchers: some of them purify water making ``tassels``, others separate them and some of them even participate in the elimination of the first. This research study is described into details and will probably be used in the future at the industrial scale. (O.M.)

  19. HPLC separation of human serum albumin isoforms based on their isoelectric points.

    Science.gov (United States)

    Turell, Lucía; Botti, Horacio; Bonilla, Lucía; Torres, María José; Schopfer, Francisco; Freeman, Bruce A; Armas, Larissa; Ricciardi, Alejandro; Alvarez, Beatriz; Radi, Rafael

    2014-01-01

    Human serum albumin (HSA) is the most abundant protein in plasma. Cys34, the only free Cys residue, is the predominant plasma thiol and a relevant sacrificial antioxidant. Both in vivo circulating HSA and pharmaceutical preparations are heterogeneous with respect to the oxidation state of Cys34. In this work, we developed an external pH gradient chromatofocusing procedure that allows the analysis of the oxidation status of HSA in human plasma and biopharmaceutical products based on the different apparent isoelectric points and chemical properties of the redox isoforms. Specifically, reduced-mercury blocked HSA (HSA-SHg(+)), HSA with Cys34 oxidized to sulfenic acid (HSA-SOH) and HSA oxidized to sulfinate anion (HSA-SO2(-)) can be separated with resolutions of 1.4 and 3.1 (first and last pair) and hence quantified and purified. In addition, an N-terminally degraded isoform (HSA3-585) in different redox states can be resolved as well. Confirmation of the identity of the chromatofocusing isolated isoforms was achieved by high resolution whole protein MS. It is proposed that the chromatofocusing procedure can be used to produce more exact and complete descriptions of the redox status of HSA in vivo and in vitro. Finally, the scalability capabilities of the chromatofocusing procedure allow for the preparation of highly pure standards of several redox isoforms of HSA. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Production of raw-starch-digesting α-amylase isoform from Bacillus sp. under solid-state fermentation and biochemical characterization.

    Science.gov (United States)

    Božić, Nataša; Slavić, Marinela Šokarda; Gavrilović, Anja; Vujčić, Zoran

    2014-07-01

    α-Amylase production by solid-state fermentation of different Bacillus sp. was studied previously on different fermentation media. However, no study has been reported on the influence of selected media on expression of desired amylase isoforms such as raw-starch-digesting amylase (RSDA). In this paper, the influence of different inexpensive and available agro-resources as solid media (corn, wheat and triticale) on α-amylase isoform induction from three wild-type Bacillus sp., selected among one hundred strains tested, namely 9B, 12B and 24A was investigated. For all three strains, tested amylases were detected in the multiple forms; however, number and intensity of each form differed depending on the solid media used for growth. To determine which isoform from Bacillus sp. 12B was RSDA, the suspected isoform was purified. The optimum pH for the purified α-amylase isoform was 6.0-8.0, while the optimum temperature was 60-90 °C. Isoform was considerably thermostable and Ca(2+)-independent, and actually the only α-amylase active towards raw starch. Purification and characterization of RSDA showed that not all of the solid media tested induced RSDA. From an economic point of view, it might be significant to obtain pure isoenzyme for potential use in the raw-starch hydrolysis, since it was 5 times more efficient in raw corn starch hydrolysis than the crude amylase preparation.

  1. [Isolation of isoforms of apolipoprotein CIII from human serum by chromatofocusing].

    Science.gov (United States)

    Fang, D; Gong, R; O, K

    1999-03-01

    This study aimed to isolate isoforms of apolipoprotein (apo) C III from human serum. 24-hour fasting serum from normal and hyperlipidemic subjects was pooled and subjected to ultracentrifugation at plasma density for 20 hours. Very low density lipoprotein (VLDL) was collected at density of d < 1.006 g/ml, and it was delipidated by ethanol and ether. The delipidated apo-VLDL was dissolved in a solution containing 7.2 mol/L urea and 20 mmol/L dithiothreitol. The insoluble apo B was removed by centrifugation. The soluble apo-VLDL was applied to PBE94 column, and eluted with elution buffer containing polybuffer 74 and 8 mol/L urea (1:8, pH4.0). After pooled, the eluted peaks of apolipoproteins were applied to column chromatography of hydroxylapatite to remove the polybuffer. The purified isoforms of apoC III and the purified apo C I, C II and E, were characterized by isoelectrofocusing and west blot. The results showed that the purified apoC III1, C III2, and C II were pure.

  2. A hardware implementation of Region-of-Interest selection in LAr-TPC for data reduction and triggering

    Energy Technology Data Exchange (ETDEWEB)

    Baibussinov, B; Centro, S; Cieslik, K; Dequal, D; Farnese, C; Fava, A; Gibin, D; Guglielmi, A; Meng, G; Pietropaolo, F; Scantamburlo, E; Varanini, F; Ventura, S [INFN - Sezione di Padova, Via Marzolo 8 35131 Padova (Italy); Rubbia, C, E-mail: francesco.pietropaolo@pd.infn.it [Laboratori Nazionali del Gran Sasso dell' INFN, I-67010 Assergi (Italy)

    2010-12-15

    Large Liquid Argon TPC detectors in the range of multi-kton mass for neutrino and astroparticle physics require the extraction and treatment of signals from some 10{sup 5} wires. In order to increase the throughput of the DAQ system an on-line lossless data compression has been realized reducing by almost a factor 4 the data flow. Moreover a new efficient on-line identification algorithm of wire hits was studied, implemented on the ICARUS digital read-out boards and fully tested on the ICARINO LAr-TPC facility operated at LNL INFN Laboratory with cosmic-rays. This system permits the extraction of the event Region-of-Interest maximizing the global throughput of the DAQ and the realization of a trigger based on charge deposition on the wires. Capability to trigger isolated low energy events down to few MeV visible energy was also demonstrated.

  3. Experimental observation of an extremely high electron lifetime with the ICARUS-T600 LAr-TPC

    CERN Document Server

    Antonello, M; Benetti, P; Boffelli, F; Bubak, A; Calligarich, E; Centro, S; Cesana, A; Cieslik, K; Cline, D B; Cocco, A G; Dabrowska, A; Dermenev, A; Dolfini, R; Falcone, A; Farnese, C; Fava, A; Ferrari, A; Fiorillo, G; Gibin, D; Gninenko, S; Guglielmi, A; Haranczyk, M; Holeczek, J; Kirsanov, M; Kisiel, J; Kochanek, I; Lagoda, J; Mania, S; Menegolli, A; Meng, G; Montanari, C; Otwinowski, S; Picchi, P; Pietropaolo, F; Plonski, P; Rappoldi, A; Raselli, G L; Rossella, M; Rubbia, C; Sala, P; Scaramelli, A; Segreto, E; Sergiampietri, F; Stefan, D; Sulej, R; Szarska, M; Terrani, M; Torti, M; Varanini, F; Ventura, S; Vignoli, C; Wang, H; Yang, X; Zalewska, A; Zani, A; Zaremba, K

    2014-01-01

    The ICARUS T600 detector, the largest liquid Argon Time Projection Chamber (LAr-TPC) realized after many years of RD activities, was installed and successfully operated for 3 years at the INFN Gran Sasso underground Laboratory. One of the most important issues was the need of an extremely low residual electronegative impurity content in the liquid Argon, in order to transport the free electrons created by the ionizing particles with a very small attenuation along the drift path. The solutions adopted for the Argon re-circulation and purification systems have permitted to reach impressive results in terms of Argon purity and a free electron lifetime exceeding 15 ms, corresponding to about 20 parts per trillion of equivalent O2 contamination, a milestone for any future project involving LAr-TPC's and the development of higher detector mass scales.

  4. A hardware implementation of Region-of-Interest selection in LAr-TPC for data reduction and triggering

    CERN Document Server

    Baibussinov, B; Cieslik, K.; Dequal, D.; Farnese, C.; Fava, A.; Gibin, D.; Guglielmi, A.; Meng, G.; Pietropaolo, F.; Rubbia, C.; Scantamburlo, E.; Varanini, F.; Ventura, S.; 10.1088/1748-0221/5/12/P12006

    2010-01-01

    Large Liquid Argon TPC detectors in the range of multikton mass for neutrino and astroparticle physics require the extraction and treatment of signals from some 105 wires. In order to enlarge the throughtput of the DAQ system an on-line lossless data compression has been realized reducing almost a factor 4 the data flow. Moreover a trigger system based on a new efficient on-line identification algorithm of wire hits was studied, implemented on the actual ICARUS digital read- out boards and fully tested on the ICARINO LAr-TPC facility operated at LNL INFN Laboratory with cosmic-rays. Capability to trigger isolated low energy events down to 1 MeV visible energy was also demonstrated.

  5. TPC码在PCM-FM遥测系统中的性能仿真%Performance simulation of TPC code in PCM-FM telemetry system

    Institute of Scientific and Technical Information of China (English)

    张定云; 向冰

    2012-01-01

    Using Matlab to build PCM-FM telemetry system and TPC code simulation model. The performance simulation of TPC code in PCM-FM telemetry system is introduced. The simulation result shows that the TPC code of code-rate is 0. 5 can gain 5 Db. Parameters of TPC decoding are simulated in this paper.%利用Matlab搭建PCM-FM遥测系统和TPC编解码仿真模型,通过仿真验证在PCM-FM遥测系统中使用TPC码后对系统的误码率改善情况.由仿真结果可知,在TPC码码率为0.5时可获得5 dB左右的增益,同时还对影响TPC增益的各个解码参数进行了详细仿真.

  6. A crustin isoform from black tiger shrimp, Penaeus monodon exhibits broad spectrum anti-bacterial activity

    Directory of Open Access Journals (Sweden)

    Debashis Banerjee

    2015-11-01

    Full Text Available Crustaceans have a powerful non-specific immune mechanism that responds to pathogen invasion and together with cellular responses, generates powerful humoral factors such as antimicrobial peptides. Crustins are a diverse class of antimicrobial peptides that are expressed by the circulating haemocytes of crustaceans. Several isoforms of this molecule are reported and in this study, one isoform from the black tiger shrimp, Penaeus monodon was cloned and expressed in Escherichia coli SG 13009. The purified recombinant crustin peptide had a molecular weight of 22 kDa and exhibited potent anti-bacterial activity in vitro against several Gram positive and Gram negative bacteria that included pathogens of aquatic animals and humans. The recombinant crustin showed a minimal inhibitory concentration of 0.5 μg ml−1 against the vibrio pathogens of shrimp, which suggests its promise for application in aquaculture.

  7. Characterization of a medium size Xe/TMA TPC instrumented with microbulk Micromegas, using low-energy \\gamma-rays

    CERN Document Server

    Alvarez, V; Carcel, S; Castel, J; Cebrian, S; Cervera, A; Conde, C A N; Dafni, T; Dias, T H V T; Diaz, J; Egorov, M; Esteve, R; Evtoukhovitch, P; Fernandes, L M P; Ferrario, P; Ferreira, A L; Freitas, E D C; Gehman, V M; Gil, A; Goldschmidt, A; Gomez, H; Gomez-Cadenas, J J; Gonzalez-Diaz, D; Gutierrez, R M; Hauptman, J; Morata, J A Hernando; Herrera, D C; Iguaz, F J; Irastorza, I G; Jinete, M A; Labarga, L; Laing, A; Liubarsky, I; Lopes, J A M; Lorca, D; Losada, M; Luzon, G; Mari, A; Martin-Albo, J; Martinez, A; Martinez-Lema, G; Miller, T; Moiseenko, A; Monrabal, F; Monteiro, C M B; Mora, F J; Moutinho, L M; Vidal, J Munoz; da Luz, H Natal; Navarro, G; Nebot-Guinot, M; Nygren, D; Oliveira, C A B; Palma, R; Perez, J; Aparicio, J L Perez; Renner, J; Ripoll, L; Rodriguez, A; Rodriguez, J; Santos, F P; Santos, J M F dos; Segui, L; Serra, L; Shuman, D; Simon, A; Sofka, C; Sorel, M; Toledo, J F; Tomas, A; Torrent, J; Tsamalaidze, Z; Vazquez, D; Veloso, J F C A; Villar, J A; Webb, R C; White, J T; Yahlali, N; Aznar, F; Calvet, D; Druillole, F; Ferrer-Ribas, E; Garcia, J A; Giomataris, I; Gracia, J; Coguie, A Le; Mols, J P; Pons, P; Ruiz, E

    2013-01-01

    NEXT-MM is a general-purpose high pressure (10 bar, $\\sim25$ l active volume) Xenon-based TPC, read out in charge mode with an 8 cm $\\times$8 cm-segmented 700 cm$^2$ plane (1152 ch) of the latest microbulk-Micromegas technology. It has been recently commissioned at University of Zaragoza as part of the R&D of the NEXT $0\

  8. A prototype of a directional detector for non-baryonic dark matter search: MIMAC (Micro-TPC Matrix of Chambers)

    Energy Technology Data Exchange (ETDEWEB)

    Grignon, C; Bernard, G; Billard, J; Bosson, G; Bourrion, O; Guillaudin, O; Koumeir, C; Mayet, F; Santos, D [LPSC, Universite Joseph Fourier Grenoble 1, CNRS/IN2P3, Institut Polytechnique de Grenoble, 53 avenue des Martyrs, 38026 Grenoble (France); Colas, P; Ferrer, E; Giomataris, I [IRFU/DSM/CEA, CEA Saclay, 91191 Gif-sur-Yvette Cedex (France); Allaoua, A; Lebreton, L, E-mail: cyril.grignon@lpsc.in2p3.f [IRSN 13115 Saint-Paul-Lez-Durance (France)

    2009-11-15

    We have developed a micro-tpc using a pixelized bulk micromegas coupled to dedicated acquisition electronics as a read-out allowing to reconstruct the three dimensional track of a few keV recoils. The prototype has been tested with the Amande facility at the IRSN-Cadarache providing monochromatic neutrons. The first results concerning discrimination of a few keV electrons and proton recoils are presented.

  9. A prototype of a directional detector for non-baryonic dark matter search: MIMAC (Micro-TPC Matrix of Chambers)

    CERN Document Server

    Grignon, C; Billard, J; Bosson, G; Bourrion, O; Guillaudin, O; Koumeir, C; Mayet, F; Santos, D; Colas, P; Ferrer, E; Giomataris, Yu; Allaoua, A; Lebreton, L

    2009-01-01

    We have developed a micro-tpc using a pixelized bulk micromegas coupled to dedicated acquisition electronics as a read-out allowing to reconstruct the three dimensional track of a few keV recoils. The prototype has been tested with the Amande facility at the IRSN-Cadarache providing monochromatic neutrons. The first results concerning discrimination of a few keV electrons and proton recoils are presented.

  10. TREX-DM: a low background Micromegas-based TPC for low mass WIMP detection

    CERN Document Server

    Iguaz, F J; Aznar, F; Castel, J F; Cebrian, S; Dafni, T; Garcia, J A; Gomez, H; Gonzalez-Diaz, D; Irastorza, I G; Lagraba, A; Luzon, G; Peiro, A; Rodriguez, A

    2015-01-01

    Dark Matter experiments are recently focusing their detection techniques in low-mass WIMPs, which requires the use of light elements and low energy threshold. In this context, we present the TREX-DM experiment, a low background Micromegas-based TPC for low-mass WIMP detection. Its main goal is the operation of an active detection mass $\\sim$0.300 kg, with an energy threshold below 0.4 keVee and fully built with previously selected radiopure materials. This article describes the actual setup, the first results of the comissioning in Ar+2\\%iC$_4$H$_{10}$ at 1.2 bar and the future updates for a possible physics run at the Canfranc Underground Laboratory in 2016. A first background model is also presented, based on Geant4 simulations and a muon/electron discrimination method. In a conservative scenario, TREX-DM could be sensitive to DAMA/LIBRA and other hints of positive WIMPs signals, with some space for improvement with a neutron/electron discrimination method or the use of other light gases.

  11. TREX-DM: a low background Micromegas-based TPC for low-mass WIMP detection

    CERN Document Server

    Iguaz, F J; Aznar, F; Castel, J F; Cebrian, S; Dafni, T; Garcia, J A; Irastorza, I G; Lagraba, A; Luzon, G; Peiro, A

    2016-01-01

    Dark Matter experiments are recently focusing their detection techniques in low-mass WIMPs, which requires the use of light elements and low energy threshold. In this context, we describe the TREX-DM experiment, a low background Micromegas-based TPC for low-mass WIMP detection. Its main goal is the operation of an active detection mass $\\sim$0.3 kg, with an energy threshold below 0.4 keVee and fully built with previously selected radiopure materials. This work describes the commissioning of the actual setup situated in a laboratory on surface and the updates needed for a possible physics run at the Canfranc Underground Laboratory (LSC) in 2016. A preliminary background model of TREX-DM is also presented, based on a Geant4 simulation, the simulation of the detector's response and two discrimination methods: a conservative muon/electron and one based on a neutron source. Based on this background model, TREX-DM could be competitive in the search for low-mass WIMPs. In particular it could be sensitive, e.g., to t...

  12. ArgonCube: a Modular Approach for Liquid Argon TPC Neutrino Detectors for Near Detector Environments

    CERN Document Server

    Auger, M; Sinclair, JR

    2017-01-01

    Liquid Argon Time Projection Chambers (LAr TPCs) are an ideal detector candidate for future neutrino oscillation physics experiments, underground neutrino observatories and proton decay searches. A large international project based on this technology is currently under consideration at the future LBNF/DUNE facility in the United States. That particular endeavor would be on the very large mass scale of 40~kt. Following diverse and long standing R\\&D work conducted over several years, with contributions from international collaborators, we propose a novel LAr TPC based on a fully-modular, innovative design, ArgonCube. ArgonCube will demonstrate that LAr TPCs are a viable detector technology for high-energy and high-multiplicity environments, such as the DUNE near detector. Necessary R\\&D work is proceeding along two main pathways; the first, aimed at the demonstration of modular detector design and the second, at the exploration of new signal readout methods. This two-pronged approach has provided a hig...

  13. GEM-based TPC with CCD Imaging for Directional Dark Matter Detection

    CERN Document Server

    Phan, N S; Lee, E R; Loomba, D; Matthews, J A J; Miller, E H

    2015-01-01

    Directional dark matter detection will require scale-ups to large volumes if low-pressure gas Time Projection Chambers (TPCs) are the only viable technology. We discuss some of the challenges for this technology, where balancing the goal of achieving the best sensitivity with that of cost effective scale-up requires an optimization over a large parameter space. Critical for this are the precision measurements of the fundamental properties of both electron and nuclear recoil tracks down to the lowest energies. Such measurements would provide a benchmark for background discrimination and directional sensitivity that could be used for future optimization studies for directional dark matter experiments. In this paper we describe a small, high resolution, high signal-to-noise GEM-based TPC with a 2D CCD readout designed for this goal. The performance of the detector was characterized using X-rays, gamma-rays, and neutrons, enabling detailed measurements of electron and nuclear recoil tracks. Stable effective gas g...

  14. Analysis of the liquid argon purity in the ICARUS T600 TPC

    Energy Technology Data Exchange (ETDEWEB)

    Amoruso, S.; Antonello, M.; Aprili, P.; Arneodo, F.; Badertscher, A.; Baiboussinov, B.; Baldo Ceolin, M.; Battistoni, G.; Bekman, B.; Benetti, P.; Bernardini, E.; Bischofberger, M.; Borio di Tigliole, A.; Brunetti, R.; Bruzzese, R.; Bueno, A.; Buzzanca, M.; Calligarich, E.; Campanelli, M.; Carbonara, F.; Carpanese, C.; Cavalli, D.; Cavanna, F.; Cennini, P.; Centro, S.; Cesana, A.; Chen, C.; Chen, D.; Chen, D.B.; Chen, Y.; Cieslik, X.; Cline, D.; Cocco, A.G.; Dai, Z.; De Vecchi, C.; Dabrowska, A.; Di Cicco, A.; Dolfini, R.; Ereditato, A.; Felcini, M.; Ferrari, A.; Ferri, F.; Fiorillo, G.; Galli, S.; Ge, Y.; Gibin, D.; Gigli Berzolari, A.; Gil-Botella, I.; Graczyk, K.; Grandi, L.; Guglielmi, A.; He, K.; Holeczek, J.; Huang, X.; Juszczak, C.; Kielczewska, D.; Kisiel, J.; Kozlowski, T.; Laffranchi, M.; Lagoda, J.; Li, Z.; Lu, F.; Ma, J.; Mangano, G.; Markiewicz, M.; Martinez de la Ossa, A.; Matthey, C.; Mauri, F.; Meng, G.; Messina, M.; Montanari, C.; Muraro, S.; Navas-Concha, S. E-mail: navas@ugr.es; Nurzia, G.; Otwinowski, S.; Ouyang, Q.; Palamara, O.; Pascoli, D.; Periale, L.; Piano Mortari, G.B.; Piazzoli, A.; Picchi, P.; Pietropaolo, F.; Polchlopek, W.; Rancati, T.; Rappoldi, A.; Raselli, G.L.; Rico, J.; Rondio, E.; Rossella, M.; Rubbia, A.; Rubbia, C.; Sala, P.; Santorelli, R.; Scannicchio, D.; Segreto, E.; Seo, Y.; Sergiampietri, F.; Sobczyk, J.; Spinelli, N.; Stepaniak, J.; Szarska, M.; Szeptycka, M.; Szleper, M.; Terrani, M.; Velotta, R.; Ventura, S.; Vignoli, C.; Wang, H.; Wang, X.; Woo, J.; Xu, G.; Xu, Z.; Zalewska, A.; Zalipska, J.; Zhang, C.; Zhang, Q.; Zhen, S.; Zipper, W

    2004-01-01

    The results reported in this paper are based on the analysis of the data recorded with the first half-module of the ICARUS T600 liquid argon Time Projection Chamber (LAr TPC), during a technical run that took place on surface in Pavia (Italy). We include results from the linearity, uniformity and calibration of the electronics, measurements on the electron drift velocity in LAr at different electric fields, as well as the LAr purity achievement of the detector. Two complementary techniques were used to measure the drift electron lifetime inside the active volume: the first, from the data of a purity monitor, gives a measurement localized in space; the second, based on the study of the signals produced by long minimum ionizing tracks crossing the detector, provides a LAr volume averaged value. Both methods yield consistent results over the whole data taking period and are compatible with an uniform LAr purity over the whole volume. The maximal drift electron lifetime value was recorded before the run stop and was about 1.8 ms. From an interpretation of the observed drift electron lifetime as a function of time, we conclude that the adopted technology would allow for drift distances exceeding 3 m.

  15. Micromegas-TPC operation at high pressure in Xenon-trimethylamine mixtures

    CERN Document Server

    Herrera, D C; Dafni, T; Ferrer-Ribas, E; Giomataris, I; Gonzalez-Diaz, D; Gómez, H; Iguaz, F J; Irastorza, I G; Luzon, G; Rodríguez, A; Segui, L; Tomás, A

    2013-01-01

    We present in this work measurements performed with a small Micromegas-TPC using a xenon-trimethylamine (Xe-TMA) Penning-mixture as filling gas. Measurements of gas gain and energy resolutions for 22.1 keV X-rays are presented, spanning several TMA concentrations and pressures between 1 and 10 bar. Across this pressure range, the best energy resolution and largest increase in gain at constant field (a standard figure for characterizing Penning-like energy transfers) is observed to be in the 1.5%-2.5% TMA region. A gain increase (at constant field) up to a factor 100 and a best energy resolution improved by up to a factor 3 with respect to the one previously reported in pure Xe -operated Micromegas, can be obtained. In virtue of the VUV-quenching properties of the mixture, the overall maximum gain achievable is also notably increased (up to 400 at 10bar), a factor x 3 higher than in pure Xe. In addition, preliminary measurements of the electron drift velocity in a modified setup have been performed and show go...

  16. Michel Electron Reconstruction Using Cosmic-Ray Data from the MicroBooNE LArTPC

    Energy Technology Data Exchange (ETDEWEB)

    Acciarri, R.; et al.

    2017-04-10

    The MicroBooNE liquid argon time projection chamber (LArTPC) has been taking data at Fermilab since 2015 collecting, in addition to neutrino beam, cosmic-ray muons. Results are presented on the reconstruction of Michel electrons produced by the decay at rest of cosmic-ray muons. Michel electrons are abundantly produced in the TPC, and given their well known energy spectrum can be used to study MicroBooNE's detector response to low-energy electrons (electrons with energies up to ~50 MeV). We describe the fully-automated algorithm developed to reconstruct Michel electrons, with which a sample of ~14,000 Michel electron candidates is obtained. Most of this article is dedicated to studying the impact of radiative photons produced by Michel electrons on the accuracy and resolution of their energy measurement. In this energy range, ionization and bremsstrahlung photon production contribute similarly to electron energy loss in argon, leading to a complex electron topology in the TPC. By profiling the performance of the reconstruction algorithm on simulation we show that the ability to identify and include energy deposited by radiative photons leads to a significant improvement in the energy measurement of low-energy electrons. The fractional energy resolution we measure improves from over 30% to ~20% when we attempt to include radiative photons in the reconstruction. These studies are relevant to a large number of analyses which aim to study neutrinos by measuring electrons produced by $\

  17. DarkSide-20k: A 20 Tonne Two-Phase LAr TPC for Direct Dark Matter Detection at LNGS

    Energy Technology Data Exchange (ETDEWEB)

    Aalseth, C.E.; et al.

    2017-07-25

    Building on the successful experience in operating the DarkSide-50 detector, the DarkSide Collaboration is going to construct DarkSide-20k, a direct WIMP search detector using a two-phase Liquid Argon Time Projection Chamber (LArTPC) with an active (fiducial) mass of 23 t (20 t). The DarkSide-20k LArTPC will be deployed within a shield/veto with a spherical Liquid Scintillator Veto (LSV) inside a cylindrical Water Cherenkov Veto (WCV). Operation of DarkSide-50 demonstrated a major reduction in the dominant $^{39}$Ar background when using argon extracted from an underground source, before applying pulse shape analysis. Data from DarkSide-50, in combination with MC simulation and analytical modeling, shows that a rejection factor for discrimination between electron and nuclear recoils of $\\gt3\\times10^9$ is achievable. This, along with the use of the veto system, is the key to unlocking the path to large LArTPC detector masses, while maintaining an "instrumental background-free" experiment, an experiment in which less than 0.1 events (other than $\

  18. Why CLEAN when you can PURIFY?

    CERN Document Server

    Carrillo, Rafael E; Wiaux, Yves

    2013-01-01

    We extend previously proposed radio-interferometric imaging approaches based on convex optimization to handle continuous visibilities and large-scale optimization problems. We propose a general algorithmic framework based on the simultaneous-direction method of multipliers to solve sparse imaging problems. The algorithm offers a parallel implementation structure, thus providing a significant gain in terms of speed and scalability to very high dimensions. We implement various state-of-the-art sparsity regularization priors, including our recent average sparsity approach SARA, in a new imaging software dubbed PURIFY. We evaluate through realistic simulations the performance of the software in terms of reconstruction quality and computational speed. Simulation results confirm both the superiority of SARA for continuous Fourier measurements and the fact that the new algorithmic structure offers a promising path to handle large-scale problems. Code is available at https://github.com/basp-group/purify

  19. Antiangiogenic VEGF Isoform in Inflammatory Myopathies

    Directory of Open Access Journals (Sweden)

    Nila Volpi

    2013-01-01

    Full Text Available Objective. To investigate expression of vascular endothelial growth factor (VEGF antiangiogenic isoform A-165b on human muscle in idiopathic inflammatory myopathies (IIM and to compare distribution of angiogenic/antiangiogenic VEGFs, as isoforms shifts are described in other autoimmune disorders. Subjects and Methods. We analyzed VEGF-A165b and VEGF-A by western blot and immunohistochemistry on skeletal muscle biopsies from 21 patients affected with IIM (polymyositis, dermatomyositis, and inclusion body myositis and 6 control muscle samples. TGF-β, a prominent VEGF inductor, was analogously evaluated. Intergroup differences of western blot bands density were statistically examined. Endomysial vascularization, inflammatory score, and muscle regeneration, as pathological parameters of IIM, were quantitatively determined and their levels were confronted with VEGF expression. Results. VEGF-A165b was significantly upregulated in IIM, as well as TGF-β. VEGF-A was diffusely expressed on unaffected myofibers, whereas regenerating/atrophic myofibres strongly reacted for both VEGF-A isoforms. Most inflammatory cells and endomysial vessels expressed both isoforms. VEGF-A165b levels were in positive correlation to inflammatory score, endomysial vascularization, and TGF-β. Conclusions. Our findings indicate skeletal muscle expression of antiangiogenic VEGF-A165b and preferential upregulation in IIM, suggesting that modulation of VEGF-A isoforms may occur in myositides.

  20. Isolation of dipeptidyl peptidase IV (DP 4) isoforms from porcine kidney by preparative isoelectric focusing to improve crystallization.

    Science.gov (United States)

    Wagner, Leona; Wermann, Michael; Rosche, Fred; Rahfeld, Jens-Ulrich; Hoffmann, Torsten; Demuth, Hans-Ulrich

    2011-07-01

    Abstract In the present studies we resolved the post-translational microheterogeneity of purified porcine dipeptidyl peptidase IV (DP 4) from kidney cortex. Applying SDS-homogeneous DP 4 onto an analytical agarose isoelectric focusing (IEF) gel, pH 4-6, activity staining resulted in at least 17 isoforms between pH 4.8-6.0. These could be separated into fractions with only two to six isoforms by means of preparative liquid-phase IEF, using a Rotofor cell. Starting off with three parallel Rotofor runs under the same conditions at pH 5-6, the fractions were pooled according to the specific activity of DP 4, pH and analytical IEF profile, and further refractionated without any additional ampholytes. Since excessive dilution of ampholytes and proteins was kept to the minimum, a second refractionation step could be introduced, resulting in pH gradients between 0.022 and 0.028 pH increments per fraction. By performing two consecutive refractionation steps, the high resolution necessary for the separation of DP 4 isoforms could be achieved. This represents an alternative method if isolation of isoforms with similar pI's results in precipitation and denaturation in presence of a narrow pH range. Furthermore, it demonstrates that preparative IEF is a powerful tool to resolve post-translational microheterogeneity of a purified protein required for crystallization processing.

  1. Molecular cloning and biochemical characterization of three phosphoglycerate kinase isoforms from developing sunflower (Helianthus annuus L.) seeds.

    Science.gov (United States)

    Troncoso-Ponce, M A; Rivoal, J; Venegas-Calerón, M; Dorion, S; Sánchez, R; Cejudo, F J; Garcés, R; Martínez-Force, E

    2012-07-01

    Three cDNAs encoding different phosphoglycerate kinase (PGK, EC 2.7.2.3) isoforms, two cytosolic (HacPGK1 and HacPGK2) and one plastidic (HapPGK), were cloned and characterized from developing sunflower (Helianthus annuus L.) seeds. The expression profiles of these genes showed differences in heterotrophic tissues, such as developing seeds and roots, where HacPGK1 was predominant, while HapPGK was highly expressed in photosynthetic tissues. The cDNAs were expressed in Escherichia coli, and the corresponding proteins purified to electrophoretic homogeneity, using immobilized metal ion affinity chromatography, and biochemically characterized. Despite the high level of identity between sequences, the HacPGK1 isoform showed strong differences in terms of specific activity, temperature stability and pH sensitivity in comparison to HacPGK2 and HapPGK. A polyclonal immune serum was raised against the purified HacPGK1 isoform, which showed cross-immunoreactivity with the other PGK isoforms. This serum allowed the localization of high expression levels of PGK isozymes in embryo tissues.

  2. A new approach to quantitate carbohydrate-deficient transferrin isoforms in alcohol abusers: partial iron saturation in isoelectric focusing/immunoblotting and laser densitometry.

    Science.gov (United States)

    Bean, P; Peter, J B

    1993-12-01

    Carbohydrate-deficient transferrin (Tf) represents a significant advance over previous markers of alcohol abuse. Isoelectric focusing (IEF) analysis of affinity-purified Tf, under conditions of total iron saturation, identifies a major isoform at pI 5.4 in both normal consumers and alcohol abusers; three additional Tf isoforms (pI 5.6, 5.7, and 5.8) are associated with alcohol abuse. Under conditions of partial iron saturation, IEF analysis of affinity-purified Tf reveals up to seven isoforms (pI range 5.3-6.0) common to normal consumers and alcohol abusers; three additional transferrin isoforms (pI range 6.1-6.3) are present in 68% (15/22) of the alcohol abuser specimens, but in only 8% (1/12) of the specimens from normal consumers and in none of the three specimens from abstainers. These three diagnostic bands comigrate with a set of defined Tf isoforms: human iron-free Tf containing two sialic acid residues, human sialic acid-free Tf with one iron molecule, and human sialic acid-free, iron-free Tf. Serum specimens from normal consumers and alcohol abusers, analyzed for Tf isoforms by an IEF-immunoblot method under conditions of partial iron saturation, expressed Tf isoforms similar to those found using affinity-purified Tf in standard IEF. Visual examination of the immunoblots reveals the diagnostic bands in 67% (32/48) of patients with histories of sustained alcohol abuse compared with only 17% (8/48) of the normal consumers. Scanning densitometry and volume integration analysis of the immunoblots representative of normal consumer and alcohol abuser populations results in mean (+/- SE) values of 4.1 +/- 0.8 and 19.3 +/- 3.6 units, respectively (p < 0.0002).

  3. Steroidogenesis in amlodipine treated purified Leydig cells

    Energy Technology Data Exchange (ETDEWEB)

    Latif, Rabia, E-mail: rabialatif08@hotmail.com [Department of Physiology, Army Medical College, National University of Sciences and Technology, Islamabad (Pakistan); Lodhi, Ghulam Mustafa, E-mail: drmustafa786@gmail.com [Department of Physiology, Wah Medical College, Wah (Pakistan); Hameed, Waqas, E-mail: waqham@hotmail.com [Department of Physiology, Rehman Medical College, Peshawar (Pakistan); Aslam, Muhammad, E-mail: professormaslam@yahoo.com [Department of Physiology, Shifa College of Medicine, Islamabad (Pakistan)

    2012-01-01

    Drugs have been shown to adversely affect male fertility and recently anti-hypertensive drugs were added to the list. The anti-fertility effects of amlodipine, a calcium channel blocker, are well-illustrated in in vivo experiments but lack an in vitro proof. The present study was designed to experimentally elucidate the effects of amlodipine on Leydig cell steroidogenesis and intracellular calcium in vitro. Leydig cells of Sprague–Dawley rats were isolated and purified by Percoll. Cells were incubated for 3 h with/without amlodipine in the presence/absence of LH, dbcAMP, Pregnenolone and 25-Hydroxycholesterol. Cytosolic calcium was measured in purified Leydig cells by fluorometric technique. The results showed significantly reduced (P < 0.05) steroidogenesis and intracellular calcium in amlodipine exposed rats. The site of amlodipine induced steroidogenic inhibition seems to be prior to the formation of Pregnenolone at the level of StAR protein. -- Highlights: ► Inhibition of steroidogenesis in isolated and purified Leydig cells by amlodipine. ► Site of inhibition was before Pregnenolone formation, at the level of StAR protein. ► Inhibition of LH stimulated rise in cytosolic calcium by amlodipine.

  4. Utilization of purified cellulose in fiber studies.

    Science.gov (United States)

    Penner, M H; Liaw, E T

    1990-01-01

    Purified cellulose-type fiber products are widely used in experimental nutrition. Their use in a broad spectrum of studies may potentially lead to the acceptance of the misconception that the various commercially available cellulose products are equivalent. In this paper we have attempted to show that this is not the case. The comparative structural data of Table 2 and the compositional data of Olsen et al provide examples which indicate that purified cellulose preparations should not necessarily be considered equivalent. Unfortunately, our current lack of understanding of how fibers are metabolized and how they may affect specific physiological parameters makes it difficult to determine which, if any, of the measurable structural and chemical properties will be of relevance for a given in vivo study. At present, it appears that researchers utilizing/evaluating the consequences of consuming a purified cellulose-type fiber would be prudent to provide at least a limited amount of data on the properties of the cellulose preparation used in their studies. The characterization of the cellulose product may be done by a variety of methods depending on the expertise of the laboratory. The methods and results discussed in this paper provide an example of the type of information which may be obtained from an in vitro characterization of cellulose products.

  5. Some Properties of Purified and Non-purified Rumen Tissue Arginase in Cattle

    OpenAIRE

    ERİŞİR, Mine; OZAN, Sema Temizer

    1998-01-01

    Some biochemical properties of purified and non-purified rumen tissue arginase were compared. Homogenization, heating, treatment with aceton, precipitation with ammonium sulfate, dialysis, several centrifugations, gel filtration on sephadex G-200 processes were utilized in the purification procedure of the enzyme. It was found that pre-incubation temperature (60 °C) of arginase and Km (4mM) to its substrate, L-arginine, did not change before and after purification. While pre-incubation peri...

  6. Ikaros isoforms:The saga continues

    Institute of Scientific and Technical Information of China (English)

    Laura; A; Perez-Casellas; Aleksandar; Savic; Sinisa; Dovat

    2011-01-01

    Through alternate splicing,the Ikaros gene produces multiple proteins.Ikaros is essential for normal hematopoiesis and possesses tumor suppressor activity.Ikaros isoforms interact to form dimers and potentially multimeric complexes.Diverse Ikaros complexes produced by the presence of different Ikaros isoforms are hypothesized to confer distinct functions.Small dominantnegative Ikaros isoforms have been shown to inhibit the tumor suppressor activity of full-length Ikaros.Here,we describe how Ikaros activity is regulated by the coordinated expression of the largest Ikaros isoforms IK-1 and IK-H.Although IK-1 is described as full-length Ikaros,IK-H is the longest Ikaros isoform.IK-H,which includes residues coded by exon 3B (60 bp that lie between exons 3 and 4),is abundant in human but not murine hematopoietic cells.Specific residues that lie within the 20 amino acids encoded by exon 3B give IK-H DNA-binding characteristics that are distinct from those of IK-1.Moreover,IK-H can potentiate or inhibit the ability of IK-1 to bind DNA.IK-H binds to the regulatory regions of genes that are upregulated by Ikaros,but not genes that are repressed by Ikaros.Although IK-1 localizes to pericentromeric heterochromatin,IK-H can be found in both pericentromeric and non-pericentromeric locations.Anti-silencing activity of gamma satellite DNA has been shown to depend on the binding of IK-H,but not other Ikaros isoforms.The unique features of IK-H,its influence on Ikaros activity,and the lack of IK-H expression in mice suggest that Ikaros function in humans may be more complex and possibly distinct from that in mice.

  7. Air Purifiers Eliminate Pathogens, Preserve Food

    Science.gov (United States)

    2009-01-01

    NASA-funded researchers produced an ethylene reduction device for a plant growth unit. KES Science & Technology Inc., a Kennesaw, Georgia-based company specializing in sustaining perishable foods, licensed the ethylene scrubbing technology. KES partnered with Akida Holdings, of Jacksonville, Florida, which now markets the NASA-developed technology as AiroCide. According to the company, it is the only air purifier that completely destroys airborne bacteria, mold, fungi, mycotoxins, viruses, volatile organic compounds (like ethylene), and odors. What?s more, the devices have no filters that need changing and produce no harmful byproducts, such as the ozone created by some filtration systems.

  8. New isoforms of rat Aquaporin-4

    DEFF Research Database (Denmark)

    Moe, Svein Erik; Sorbo, Jan Gunnar; Søgaard, Rikke;

    2008-01-01

    an intracellular localization when expressed in cell lines and do not transport water when expressed in Xenopus oocytes. In contrast, the largest of the new isoforms, AQP4e, which contains a novel N-terminal domain, is localized at the plasma membrane in cell lines and functions as a water transporter in Xenopus...

  9. p53 isoforms change p53 paradigm

    OpenAIRE

    2014-01-01

    Although p53 defines cellular responses to cancer treatment it is not clear how p53 can be used to control cell fate outcome. Data demonstrate that so-called p53 does not exist as a single protein, but is in fact a group of p53 protein isoforms whose expression can be manipulated to control the cellular response to treatment.

  10. Isoforms of receptors of fibroblast growth factors.

    Science.gov (United States)

    Gong, Siew-Ging

    2014-12-01

    The breadth and scope of Fibroblast Growth Factor signaling is immense, with documentation of its role in almost every organism and system studied so far. FGF ligands signal through a family of four distinct tyrosine kinase receptors, the FGF receptors (FGFRs). One contribution to the diversity of function and signaling of FGFs and their receptors arises from the numerous alternative splicing variants that have been documented in the FGFR literature. The present review discusses the types and roles of alternatively spliced variants of the FGFR family members and the significant impact of alternative splicing on the physiological functions of five broad classes of FGFR isoforms. Some characterized known regulatory mechanisms of alternative splicing and future directions in studies of FGFR alternative splicing are also discussed. Presence, absence, and/or the combination of specific exons within each FGFR protein impart upon each individual isoform its unique function and expression pattern during normal function and in diseased states (e.g., in cancers and birth defects). A better understanding of the diversity of FGF signaling in different developmental contexts and diseased states can be achieved through increased knowledge of the presence of specific FGFR isoforms and their impact on downstream signaling and functions. Modern high-throughput techniques afford an opportunity to explore the distribution and function of isoforms of FGFR during development and in diseases.

  11. Study on soot purifying of molding shop in coking factory

    Institute of Scientific and Technical Information of China (English)

    LI Duo-song; ZHANG Hui; BAI Xiang-yu

    2006-01-01

    Exhaust gas in molding shop was complicated in component and characteristic in Iow thickness asphalt smoke, mass steam-gas and dust. It was difficult to purify the soot with common purifier. So we must consider them roundly and develop new multifunction purifier. PFP multifunction soot purifier was made on the base of design optimization and was installed at Shenhuo Coking Factory in 2004. The combined effects of multi- mechanism in purifier make purifying ratio keep in high level. The remove ratio of smut reaches at 92.8%, and asphalt smoke at 83.7%.

  12. Apolipoprotein (A) Isoform Distribution and Plasma Lipoprotein (a ...

    African Journals Online (AJOL)

    Apolipoprotein (A) Isoform Distribution and Plasma Lipoprotein (a) Levels In Nigerian Subjects With and Without Coronary Heart Disease. ... Plasma lipoprotein (a) Concentrations and apo(a) isoforms were determined in 101 ... Article Metrics.

  13. Long-chain acyl-CoA synthetase isoforms differ in preferences for eicosanoid species and long-chain fatty acids.

    Science.gov (United States)

    Klett, Eric L; Chen, Shufen; Yechoor, Alekhya; Lih, Fred B; Coleman, Rosalind A

    2017-02-16

    Because the signaling eicosanoids, epoxyeicosatrienoic acids (EETs) and hydroxyeicosatetraenoic acids (HETEs), are esterified to membrane phospholipids, we asked which long-chain acyl-CoA synthetase (ACSL) isoforms would activate these molecules and whether the apparent fatty acid substrate preferences of each ACSL isoform might differ depending on whether it was assayed in mammalian cell membranes or as a purified bacterial recombinant protein. We found that all five ACSL isoforms were able to use EETs and HETEs as substrates and showed by LC-MS that ACSLs produce EET-CoAs. We found differences in substrate preference between ACS assays performed in COS7 cell membranes and recombinant purified proteins. Similarly, preferences and Michaelis-Menten kinetics for long-chain fatty acids were distinctive. Substrate preferences identified for the purified ACSLs did not correspond to those observed in ACSL-deficient mouse models. Taken together, these data support the concept that each ACSL isoform exhibits a distinct substrate preference, but apparent substrate specificities depend upon multiple factors including membrane character, co-activators, inhibitors, protein interactions, and post-translational modification.

  14. 基于回归模型的高端容错计算机TPC-C性能估算研究%A Regression-Based Prediction Model for TPC-C Performance of High-End Fault-Tolerant Computers

    Institute of Scientific and Technical Information of China (English)

    刘迪; 翟季冬; 陈文光

    2013-01-01

    高端容错计算机的TPC-C性能测试由于成本高昂且时间漫长,导致市场上只有少部分产品进行了该项测试,无法满足生产商和购买者的需求,高端容错计算机领域需要一种简便快捷、低成本的TPC-C性能估算方法.文中分析了影响TPC-C性能的各种因素,以近5年来发布了TPC-C测试结果的服务器为样本,利用数理统计的方法,在服务器TPC-C性能与硬件指标之间建立了线性回归模型.优化后的模型估算精度达到95%以上,在一定程度上解释了服务器的硬件指标与TPC-C性能之间的因果关系,具备了方便准确地估算TPC-C性能的现实意义.文中所提出的将数理统计的方法用于TPC-C性能估算的思路以及搜集的大量相关数据,对今后该项研究具有重要意义.%As the TPC-C performance test of high-end fault-tolerant computer cost expensively and long time,only a small part of the products on the market do the test,which can not meet the needs of producers and buyers.A simple,efficient and low-cost method for estimating the TPC-C performance is widely needed.This paper analyzes the various factor affecting the TPC-C performance,establishes a linear regression model between the TPC-C performance and hardware indicators using the method of mathematical statistics.The model takes the TPC-C test results released in the past five years as the sample.After tuning,the estimating accuracy of the model is more than 95%.The model explains the causal relationship between the server's hardware indicators and TPC-C performance to some extent,and estimates the TPC-C performance conveniently and accurately.The idea that using mathematical statistics for TPC-C performance estimation and the large volume of data collected has important significance for future study.

  15. Gating of the two-pore cation channel AtTPC1 in the plant vacuole is based on a single voltage-sensing domain.

    Science.gov (United States)

    Jaślan, D; Mueller, T D; Becker, D; Schultz, J; Cuin, T A; Marten, I; Dreyer, I; Schönknecht, G; Hedrich, R

    2016-09-01

    The two-pore cation channel TPC1 operates as a dimeric channel in animal and plant endomembranes. Each subunit consists of two homologous Shaker-like halves, with 12 transmembrane domains in total (S1-S6, S7-S12). In plants, TPC1 channels reside in the vacuolar membrane, and upon voltage stimulation, give rise to the well-known slow-activating SV currents. Here, we combined bioinformatics, structure modelling, site-directed mutagenesis, and in planta patch clamp studies to elucidate the molecular mechanisms of voltage-dependent channel gating in TPC1 in its native plant background. Structure-function analysis of the Arabidopsis TPC1 channel in planta confirmed that helix S10 operates as the major voltage-sensing site, with Glu450 and Glu478 identified as possible ion-pair partners for voltage-sensing Arg537. The contribution of helix S4 to voltage sensing was found to be negligible. Several conserved negative residues on the luminal site contribute to calcium binding, stabilizing the closed channel. During evolution of plant TPC1s from two separate Shaker-like domains, the voltage-sensing function in the N-terminal Shaker-unit (S1-S4) vanished.

  16. Ozone Air Purifiers: Can They Improve Asthma Symptoms?

    Science.gov (United States)

    ... daughter has asthma. Would she benefit from an ozone air purifier in her room? Answers from James ... Li, M.D., Ph.D. Despite manufacturers' claims, ozone air purifiers don't remove asthma triggers from ...

  17. Soluble expression and characterization of a GFP-fused pea actin isoform(PEAc1)

    Institute of Scientific and Technical Information of China (English)

    Ai Xiao LIU; Shao Bin ZHANG; Xiao Jing XU; Dong Tao REN; Guo Qin LIU

    2004-01-01

    A pea actin isoform PEAc1 with green fluorescent protein (GFP) fusion to its C-terminus and His-tag to its Nterminus, was expressed in prokaryotic cells in soluble form, and highly purified with Ni-Chelating SepharoseTM Fast Flow column. The purified fusion protein (PEAc1-GFP) efficiently inhibited DNase I activities before polymerization,and activated the myosin Mg-ATPase activities after polymerization. The PEAc 1-GFP also polymerized into green fluorescent filamentous structures with a critical concentration of 0.75 μM. These filamentous structures were labeled by TRITC-phalloidin, a specific agent for staining actin microfilaments, and identified as having 9 nm diameters by negative staining. These results indicated that PEAc1 preserved the essential characteristics of actin even with His-tag and GFP fusion, suggesting a promising potential to use GFP fusion protein in obtainning soluble plant actin isoform to analyze its physical and biochemical properties in vitro. The PEAc 1-GFP was also expressed in tobacco BY2 cells,which offers a new pathway for further studying its distribution and function in vivo.

  18. Isolating and Purifying Clostridium difficile Spores

    Science.gov (United States)

    Edwards, Adrianne N.; McBride, Shonna M.

    2016-01-01

    Summary The ability for the obligate anaerobe, Clostridium difficile, to form a metabolically dormant spore is critical for the survival of this organism outside of the host. This spore form is resistant to a myriad of environmental stresses, including heat, desiccation and exposure to disinfectants and antimicrobials. These intrinsic properties of spores allow C. difficile to survive long-term in an oxygenated environment, to be easily transmitted from host-to-host and to persist within the host following antibiotic treatment. Because of the importance of the spore form to the C. difficile lifecycle and treatment and prevention of C. difficile infection (CDI), the isolation and purification of spores are necessary to study the mechanisms of sporulation and germination, investigate spore properties and resistances, and for use in animal models of CDI. This chapter provides basic protocols, in vitro growth conditions and additional considerations for purifying C. difficile spores for a variety of downstream applications. PMID:27507337

  19. 21 CFR 880.6710 - Medical ultraviolet water purifier.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Medical ultraviolet water purifier. 880.6710... Miscellaneous Devices § 880.6710 Medical ultraviolet water purifier. (a) Identification. A medical ultraviolet water purifier is a device intended for medical purposes that is used to destroy bacteria in water...

  20. Functional studies of sodium pump isoforms

    DEFF Research Database (Denmark)

    Clausen, Michael Jakob

    The Na+,K+-ATPase is an essential ion pump found in all animal cells. It uses the energy from ATP hydrolysis to export three Na+ and import two K+, both against their chemical gradients and for Na+ also against the electrical potential. Mammals require four Na+,K+-ATPase isoforms that each have u...... synthesized cohorts of pumps from the Golgi apparatus to the plasma membrane....

  1. FSH isoform pattern in classic galactosemia.

    Science.gov (United States)

    Gubbels, Cynthia S; Thomas, Chris M G; Wodzig, Will K W H; Olthaar, André J; Jaeken, Jaak; Sweep, Fred C G J; Rubio-Gozalbo, M Estela

    2011-04-01

    Female classic galactosemia patients suffer from primary ovarian insufficiency (POI). The cause for this long-term complication is not fully understood. One of the proposed mechanisms is that hypoglycosylation of complex molecules, a known secondary phenomenon of galactosemia, leads to FSH dysfunction. An earlier study showed less acidic isoforms of FSH in serum samples of two classic galactosemia patients compared to controls, indicating hypoglycosylation. In this study, FSH isoform patterns of five classic galactosemia patients with POI were compared to the pattern obtained in two patients with a primary glycosylation disorder (phosphomannomutase-2-deficient congenital disorders of glycosylation, PMM2-CDG) and POI, and in five postmenopausal women as controls. We used FPLC chromatofocussing with measurement of FSH concentration per fraction, and discovered that there were no significant differences between galactosemia patients, PMM2-CDG patients and postmenopausal controls. Our results do not support that FSH dysfunction due to a less acidic isoform pattern because of hypoglycosylation is a key mechanism of POI in this disease.

  2. TREX-DM: a low-background Micromegas-based TPC for low-mass WIMP detection

    Energy Technology Data Exchange (ETDEWEB)

    Iguaz, F.J.; Garza, J.G.; Castel, J.F.; Cebrian, S.; Dafni, T.; Garcia, J.A.; Irastorza, I.G.; Lagraba, A.; Luzon, G.; Peiro, A. [Universidad de Zaragoza, Grupo de Fisica Nuclear y Astroparticulas, Zaragoza (Spain); Aznar, F. [Universidad de Zaragoza, Grupo de Fisica Nuclear y Astroparticulas, Zaragoza (Spain); Universidad de Zaragoza, Centro Universitario de la Defensa, Zaragoza (Spain)

    2016-10-15

    If Dark Matter is made of Weakly Interacting Massive Particles (WIMPs) with masses below ∝20 GeV, the corresponding nuclear recoils in mainstream WIMP experiments are of energies too close, or below, the experimental threshold. Gas Time Projection Chambers (TPCs) can be operated with a variety of target elements, offer good tracking capabilities and, on account of the amplification in gas, very low thresholds are achievable. Recent advances in electronics and in novel radiopure TPC readouts, especially micro-mesh gas structure (Micromegas), are improving the scalability and low-background prospects of gaseous TPCs. Here we present TREX-DM, a prototype to test the concept of a Micromegas-based TPC to search for low-mass WIMPs. The detector is designed to host an active mass of ∝0.300 kg of Ar at 10 bar, or alternatively ∝0.160 kg of Ne at 10 bar, with an energy threshold below 0.4 keVee, and is fully built with radiopure materials. We will describe the detector in detail, the results from the commissioning phase on surface, as well as a preliminary background model. The anticipated sensitivity of this technique may go beyond current experimental limits for WIMPs of masses of 2-8 GeV. (orig.)

  3. The WA105-3x1x1 m3 dual phase LAr-TPC demonstrator

    CERN Document Server

    Murphy, Sebastien

    2016-11-15

    The dual phase Liquid Argon Time Projection Chamber (LAr TPC) is the state-of-art technology for neutrino detection thanks to its superb 3D tracking and calorimetry performance. Its main feature is the charge amplification in gas argon which provides excellent signal-to-noise ratio. Electrons produced in the liquid argon are extracted in the gas phase. Here, a readout plane based on Large Electron Multiplier detectors provides amplification of the charges before its collection onto an anode with strip readout. The charge amplification enables constructing fully homoge- nous giant LAr-TPCs with tuneable gain, excellent charge imaging performance and increased sensitivity to low energy events. Following a staged approach the WA105 collaboration is con- structing a dual phase LAr-TPC with an active volume of 3x1x1m3 that will soon be tested with cosmic rays. Its construction and operation aims to test scalable solutions for the crucial aspects of this technology: ultra high argon purity in non-evacuable tank, la...

  4. First results of a large-area cryogenic gaseous photomultiplier coupled to a dual-phase liquid xenon TPC

    CERN Document Server

    Arazi, L; Erdal, E; Israelashvili, I; Rappaport, M L; Shchemelinin, S; Vartsky, D; Santos, J M F dos; Breskin, A

    2015-01-01

    We discuss recent advances in the development of cryogenic gaseous photomultipliers (GPM), for possible use in dark matter and other rare-event searches using noble-liquid targets. We present results from a 10 cm diameter GPM coupled to a dual-phase liquid xenon (LXe) TPC, demonstrating - for the first time - the feasibility of recording both primary ("S1") and secondary ("S2") scintillation signals. The detector comprised a triple Thick Gas Electron Multiplier (THGEM) structure with cesium iodide photocathode on the first element; it was shown to operate stably at 180 K with gains above 10^5, providing high single-photon detection efficiency even in the presence of large alpha particle-induced S2 signals comprising thousands of photoelectrons. S1 scintillation signals were recorded with a time resolution of 1.2 ns (RMS). The energy resolution ({\\sigma}/E) for S2 electroluminescence of 5.5 MeV alpha particles was ~9%, which is comparable to that obtained in the XENON100 TPC with PMTs. The results are discusse...

  5. TREX-DM: a low-background Micromegas-based TPC for low-mass WIMP detection

    Science.gov (United States)

    Iguaz, F. J.; Garza, J. G.; Aznar, F.; Castel, J. F.; Cebrián, S.; Dafni, T.; García, J. A.; Irastorza, I. G.; Lagraba, A.; Luzón, G.; Peiró, A.

    2016-10-01

    If Dark Matter is made of Weakly Interacting Massive Particles (WIMPs) with masses below {˜ }20 GeV, the corresponding nuclear recoils in mainstream WIMP experiments are of energies too close, or below, the experimental threshold. Gas Time Projection Chambers (TPCs) can be operated with a variety of target elements, offer good tracking capabilities and, on account of the amplification in gas, very low thresholds are achievable. Recent advances in electronics and in novel radiopure TPC readouts, especially micro-mesh gas structure (Micromegas), are improving the scalability and low-background prospects of gaseous TPCs. Here we present TREX-DM, a prototype to test the concept of a Micromegas-based TPC to search for low-mass WIMPs. The detector is designed to host an active mass of {˜ }0.300 kg of Ar at 10 bar, or alternatively {˜ }0.160 kg of Ne at 10 bar, with an energy threshold below 0.4 keVee, and is fully built with radiopure materials. We will describe the detector in detail, the results from the commissioning phase on surface, as well as a preliminary background model. The anticipated sensitivity of this technique may go beyond current experimental limits for WIMPs of masses of 2-8 GeV.

  6. Search for anomalies in the neutrino sector with muon spectrometers and large LArTPC imaging detectors at CERN

    CERN Document Server

    Antonello, A; Baibussinov, B; Bilokon, H; Boffelli, F; Bonesini, M; Calligarich, E; Canci, N; Centro, S; Cesana, A; Cieslik, K; Cline, D B; Cocco, A G; Dequal, D; Dermenev, A; Dolfini, R; De Gerone, M; Dussoni, S; Farnese, C; Fava, A; Ferrari, A; Fiorillo, G; Garvey, G T; Gatti, F; Gibin, D; Gninenko, S; Guber, F; Guglielmi, A; Haranczyk, M; Holeczek, J; Ivashkin, A; Kirsanov, M; Kisiel, J; Kochanek, I; Kurepin, A; Lagoda, J; Lucchini, G; Louis, W C; Mania, S; Mannocchi, G; Marchini, S; Matveev, V; Menegolli, A; Meng, G; Mills, G B; Montanari, C; Nicoletto, M; Otwinowski, S; Palczewki, T J; Passardi, G; Perfetto, F; Picchi, P; Pietropaolo, F; Plonski, P; Rappoldi, A; Raselli, G L; Rossella, M; Rubbia, C; Sala, P; Scaramelli, A; Segreto, E; Stefan, D; Stepaniak, J; Sulej, R; Suvorova, O; Terrani, M; Tlisov, D; Van de Water, R G; Trinchero, G; Turcato, M; Varanini, F; Ventura, S; Vignoli, C; Wang, H G; Yang, X; Zani, A; Zaremba, K; Benettoni, M; Bernardini, P; Bertolin, A; Brugnera, R; Calabrese, M; Cecchetti, A; Cecchini, S; Collazuol, G; Creti, P; Corso, F Dal; Del Prete, A; De Mitri, I; De Robertis, G; De Serio, M; Esposti, L Degli; Di Ferdinando, D; Dore, U; Dusini, S; Fabbricatore, P; Fanin, C; Fini, R A; Fiore, G; Garfagnini, A; Giacomelli, G; Giacomelli, R; Guandalini, C; Guerzoni, M; Kose, U; Laurenti, G; Laveder, M; Lippi, I; Loddo, F; Longhin, A; Loverre, P; Mancarella, G; Mandrioli, G; Margiotta, A; Marsella, G; Mauri, N; Medinaceli, E; Mengucci, A; Mezzetto, M; Michinelli, R; Muciaccia, M T; Orecchini, D; Paoloni, A; Papadia, G; Pastore, A; Patrizii, L; Pozzato, M; Rosa, G; Sahnounm, Z; Simone, S; Sioli, M; Sirri, G; Spurio, M; Stanco, L; Surdo, A; Tenti, M; Togo, V; Ventura, M; Zago, M

    2012-01-01

    A new experiment with an intense ~2 GeV neutrino beam at CERN SPS is proposed in order to definitely clarify the possible existence of additional neutrino states, as pointed out by neutrino calibration source experiments, reactor and accelerator experiments and measure the corresponding oscillation parameters. The experiment is based on two identical LAr-TPCs complemented by magnetized spectrometers detecting electron and muon neutrino events at Far and Near positions, 1600 m and 300 m from the proton target, respectively. The ICARUS T600 detector, the largest LAr-TPC ever built with a size of about 600 ton of imaging mass, now running in the LNGS underground laboratory, will be moved at the CERN Far position. An additional 1/4 of the T600 detector (T150) will be constructed and located in the Near position. Two large area spectrometers will be placed downstream of the two LAr-TPC detectors to perform charge identification and muon momentum measurements from sub-GeV to several GeV energy range, greatly comple...

  7. Transport of platinum bonded nucleotides into proteoliposomes, mediated by Drosophila melanogaster thiamine pyrophosphate carrier protein (DmTpc1).

    Science.gov (United States)

    Carrisi, Chiara; Antonucci, Daniela; Lunetti, Paola; Migoni, Danilo; Girelli, Chiara R; Dolce, Vincenza; Fanizzi, Francesco P; Benedetti, Michele; Capobianco, Loredana

    2014-01-01

    The results of the present study suggest that DmTpc1 is actively implicated in the specific uptake of free cytoplasmic Pt bonded nucleotides, and therefore could be linked to the mechanism of action of some platinum-based antitumor drugs. Although DmTpc1 has a low affinity for model [Pt(dien)(N7-5'-dGTP)] and cis-[Pt(NH3)2(py)(N7-5'-dGTP)] compared to dATP it's well known that DNA platination level of few metal atoms per double-stranded molecule may account for the pharmacological activity of platinum based antitumor drugs. This is the first investigation where it has been demonstrated that a mitochondrial carrier is directly involved in the transport of metalated purines related with the cisplatin mechanism of action. Moreover it is shown as a lower hindrance of nucleotide bonded platinum complexes could strongly enhance mitochondrial uptake. Furthermore, a new application of ICP-AES addressed to measure the transport of metalated nucleobases, by using a recombinant protein reconstituted into liposomes, has been here, for the first time, developed and compared with a standard technique such as the liquid scintillation counting.

  8. The ALICE TPC, a large 3-dimensional tracking device with fast readout for ultra-high multiplicity events

    CERN Document Server

    INSPIRE-00249819; Appelshauser, H.; Bablok, S.; Bialas, N.; Bolgen, R.; Bonnes, U.; Bramm, R.; Braun-Munzinger, P.; Campagnolo, R.; Christiansen, P.; Dobrin, A.; Engster, C.; Fehlker, D.; Foka, Y.; Frankenfeld, U.; Gaardhoje, J.J.; Garabatos, C.; Glassel, P.; Gonzalez Gutierrez, C.; Gros, P.; Gustafsson, H.A.; Helstrup, H.; Hoch, M.; Ivanov, M.; Janik, R.; Junique, A.; Kalweit, A.; Keidel, R.; Kniege, S.; Kowalski, M.; Larsen, D.T.; Lesenechal, Y.; Lenoir, P.; Lindegaard, N.; Lippmann, C.; Mager, M.; Mast, M.; Matyja, A.; Munkejord, M.; Musa, L.; Nielsen, B.S.; Nikolic, V.; Oeschler, H.; Olsen, E.K.; Oskarsson, A.; Osterman, L.; Pikna, M.; Rehman, A.; Renault, G.; Renfordt, R.; Rossegger, S.; Rohrich, D.; Roed, K.; Richter, M.; Rueshmann, G.; Rybicki, A.; Sann, H.; Schmidt, H.R.; Siska, M.; Sitar, B.; Soegaard, C.; Soltveit, H.K.; Soyk, D.; Stachel, J.; Stelzer, H.; Stenlund, E.; Stock, R.; Strmen, P.; Szarka, I.; Ullaland, K.; Vranic, D.; Veenhof, R.; Westergaard, J.; Wiechula, J.; Windelband, B.

    2010-01-01

    The design, construction, and commissioning of the ALICE Time-Projection Chamber (TPC) is described. It is the main device for pattern recognition, tracking, and identification of charged particles in the ALICE experiment at the CERN LHC. The TPC is cylindrical in shape with a volume close to 90 m^3 and is operated in a 0.5 T solenoidal magnetic field parallel to its axis. In this paper we describe in detail the design considerations for this detector for operation in the extreme multiplicity environment of central Pb--Pb collisions at LHC energy. The implementation of the resulting requirements into hardware (field cage, read-out chambers, electronics), infrastructure (gas and cooling system, laser-calibration system), and software led to many technical innovations which are described along with a presentation of all the major components of the detector, as currently realized. We also report on the performance achieved after completion of the first round of stand-alone calibration runs and demonstrate result...

  9. Measurement of 1.7 to 74 MeV polarised gamma rays with the HARPO TPC

    CERN Document Server

    Geerebaert, Y; Amano, S; Attié, D; Bernard, D; Bruel, P; Calvet, D; Colas, P; Daté, S; Delbart, A; Frotin, M; Giebels, B; Götz, D; Hashimoto, S; Horan, D; Kotaka, T; Louzir, M; Minamiyama, Y; Miyamoto, S; Ohkuma, H; Poilleux, P; Semeniouk, I; Sizun, P; Takemoto, A; Yamaguchi, M; Wang, S

    2016-01-01

    Current {\\gamma}-ray telescopes based on photon conversions to electron-positron pairs, such as Fermi, use tungsten converters. They suffer of limited angular resolution at low energies, and their sensitivity drops below 1 GeV. The low multiple scattering in a gaseous detector gives access to higher angular resolution in the MeV-GeV range, and to the linear polarisation of the photons through the azimuthal angle of the electron-positron pair. HARPO is an R&D program to characterise the operation of a TPC (Time Projection Chamber) as a high angular-resolution and sensitivity telescope and polarimeter for {\\gamma} rays from cosmic sources. It represents a first step towards a future space instrument. A 30 cm cubic TPC demonstrator was built, and filled with 2 bar argon-based gas. It was put in a polarised {\\gamma}-ray beam at the NewSUBARU accelerator in Japan in November 2014. Data were taken at different photon energies from 1.7 MeV to 74 MeV, and with different polarisation configurations. The electronic...

  10. Optical Readout Time Projection Chamber (O-TPC) for a Study of Oxygen Formation In Stellar Helium Burning

    CERN Document Server

    Gai, M; Chechik, R; Dangendorf, V; Weller, H R; Gai, Moshe; Breskin, Amos; Chechik, Rachel; Dangendorf, Volker; Weller, Henry R.

    2005-01-01

    We are developing an Optical Readout Time Projection Chamber (O-TPC) detector for the study of the 12C(a,g)16O reaction that determines the ratio of carbon to oxygen in helium burning. This ratio is crucial for understanding the final fate of a progenitor star and the nucleosynthesis of elements prior to a Type II supernova; an oxygen rich star is predicted to collapse to a black hole, and a carbon rich star to a neutron star. Type Ia supernovae (SNeIa) are used as standard candles for measuring cosmological distances with the use of an empirical light curve-luminosity stretching factor. It is essential to understand helium burning that yields the carbon/oxygen white dwarf and thus the initial stage of SNeIa. The O-TPC is intended for use with high intensity photon beams extracted from the HIgS/TUNL facility at Duke University to study the 16O(g,a)12C reaction, and thus the direct reaction at energies as low as 0.7 MeV. We are conducting a systematical study of the best oxygen containing gas with light emitti...

  11. HARPO: beam characterization of a TPC for gamma-ray polarimetry and high angular-resolution astronomy in the MeV-GeV range

    CERN Document Server

    Wang, Shaobo; Bruel, Philippe; Frotin, Mickael; Geerebaert, Yannick; Giebels, Berrie; Gros, Philippe; Horan, Deirdre; Louzir, Marc; Poilleux, Patrick; Semeniouk, Igor; Attié, David; Calvet, Denis; Colas, Paul; Delbart, Alain; Sizun, Patrick; Götz, Diego; Amano, Sho; Kotaka, Takuya; Hashimoto, Satoshi; Minamiyama, Yasuhito; Takemoto, Akinori; Yamaguchi, Masashi; Miyamoto, Shuji; Daté, Schin; Ohkuma, Haruo

    2015-01-01

    A time projection chamber (TPC) can be used to measure the polarization of gamma rays with excellent angular precision and sensitivity in the MeV-GeV energy range through the conversion of photons to e+e- pairs. The Hermetic ARgon POlarimeter (HARPO) prototype was built to demonstrate this concept. It was recently tested in the polarized photon beam at the NewSUBARU facility in Japan. We present this data-taking run, which demonstrated the excellent performance of the HARPO TPC.

  12. Single molecule DNA compaction by purified histones

    Institute of Scientific and Technical Information of China (English)

    RAN ShiYong; WANG XiaoLing; FU WenBo; WANG WeiChi; LI Ming

    2008-01-01

    The compaction of single DNA molecules by purified histones is studied using magnetic tweezers, The compaction rate increases rapidly when the histone concentration is increased from 0.002 to 0.2 mmol/L, and saturates when the concentration is beyond 0.2 mmol/L, The time course of compaction is exponential at low histone concentrations. It becomes sigmoidal at high concentrations. Cooperativity between the histones bound to DNA is proposed to be responsible for the transition. The histones are loaded onto DNA randomly at low concentrations. They tend to bind DNA cooperatively at high con-centrations because the structural torsions of DNA induced by the bound histones become overlapping so that the binding of one histone facilitates the binding of others. Under very large forces, the com-pacted histone-DNA complex can be disrupted in a discrete manner with a step size of ~60 nm. But the histones cannot be completely stripped off DNA, as is revealed by the lowered B-S transition plateau of the histone-bound DNA.

  13. The three isoforms of the light-harvesting complex II Spectroscopic features, trimer formation, and functional roles

    CERN Document Server

    Standfuss, Jorg

    2004-01-01

    The major light-harvesting complex (LHC-II) of higher plants plays a crucial role in capturing light energy for photosynthesis and in regulating the flow of energy within the photosynthetic apparatus. Native LHC-II isolated from plant tissue consists of three isoforms, Lhcb1, Lhcb2, and Lhcb3, which form homo- and heterotrimers. All three isoforms are highly conserved among different species, suggesting distinct functional roles. We produced the three LHC-II isoforms by heterologous expression of the polypeptide in Escherichia coli and in vitro refolding with purified pigments. Although Lhcb1 and Lhcb2 are very similar in polypeptide sequence and pigment content, Lhcb3 is clearly different because it lacks an N-terminal phosphorylation site and has a higher chlorophyll a/b ratio, suggesting the absence of one chlorophyll b. Low temperature absorption and fluorescence emission spectra of the pure isoforms revealed small but significant differences in pigment organization. The oligomeric state of the pure isofo...

  14. Human invariant chain isoform p35 restores thymic selection and antigen presentation in CD74-deficient mice.

    Science.gov (United States)

    Genève, Laetitia; Chemali, Magali; Desjardins, Michel; Labrecque, Nathalie; Thibodeau, Jacques

    2012-10-01

    The invariant chain (Ii) has pleiotropic functions and is a key factor in antigen presentation. Ii associates with major histocompatibility complex class II molecules in the endoplasmic reticulum (ER) and targets the complex in the endocytic pathway to allow antigenic peptide loading. The human Iip35 isoform includes a cytoplasmic extension containing a di-arginine motif causing ER retention. This minor isoform does not exist in mice and its function in humans has not been thoroughly investigated. We have recently generated transgenic mice expressing Iip35 and these were crossed with Ii-deficient mice to generate animals (Tgp35/mIiKO) expressing exclusively the human isoform. In these mice, we show that Iip35 is expressed in antigen presenting cells and is inducible by interferon gamma (IFN-γ). Despite the low constitutive expression of the protein and some minor differences in the Vβ repertoire of Tgp35/mIiKO mice, Iip35 restored thymic selection of CD4(+) T cells and of invariant natural killer T cells. In vitro functional assays using purified primary macrophages treated with IFN-γ showed that Iip35 allows presentation of an Ii-dependent ovalbumin T-cell epitope. Altogether, our results suggest that Iip35 is functional and does not require co-expression of other isoforms for antigen presentation.

  15. Analysis of the Fragile X mental retardation protein isoforms 1, 2 and 3 interactions with the G-quadruplex forming semaphorin 3F mRNA.

    Science.gov (United States)

    Evans, Timothy L; Blice-Baum, Anna C; Mihailescu, Mihaela-Rita

    2012-02-01

    Fragile X syndrome, the most prevalent inheritable mental retardation, is caused by the loss of fragile X mental retardation protein (FMRP) expression. FMRP is an RNA-binding protein with nucleo-cytoplasmic shuttle activity, proposed to act as a translation regulator of specific mRNAs in the brain. It has been shown that FMRP uses its arginine-glycine-glycine (RGG) box domain to bind a subset of mRNA targets that form a G-quadruplex structure. FMRP has also been shown to undergo the post-translational modifications of arginine methylation and phosphorylation, as well as alternative splicing, resulting in multiple isoforms. The alternative splice isoforms investigated in this study, isoform 1 (ISO1), isoform 2 (ISO2), and isoform 3 (ISO3), are created by the alternative splicing acceptor site at exon 15. FMRP ISO2 and ISO3 are truncated by 12 and 13 residues, respectively, relative to the longest FMRP isoform ISO1. These truncations, which are in the close proximity of the RGG box domain, preserve the integrity of the RGG box in all three isoforms, but eliminate the in vivo phosphorylation sites, present only on FMRP ISO1. We have expressed and purified recombinant FMRP ISO1, ISO2 and ISO3 in Escherichia coli, free of post-translational modifications, and by using fluorescence spectroscopy, we show that each FMRP isoform binds G-quadruplex RNA, albeit with different binding affinities, suggesting that naturally occurring sequence modifications in the proximity of the RGG box modulate its G-quadruplex RNA binding ability.

  16. Expression of Contractile Protein Isoforms in Microgravity

    Science.gov (United States)

    Anderson, Page A. W.

    1996-01-01

    The general objective of this experiment is to determine the effect of space flight parameters, including microgravity, on ontogenesis and embryogenesis of Japanese quail. Nine U.S. and two Russian investigators are cooperating in this study. Specific objectives of the participating scientists include assessing the gross and microscopic morphological and histological development of the embryo, as well as the temporal and spacial development of specific cells, tissues, and organs. Temporally regulated production of specific proteins is also being investigated. Our objective is to determine the effects of microgravity on developmentally programmed expression of Troponin T and I isoforms known to regulate cardiac and skeletal muscle contraction.

  17. HARPO—A gaseous TPC for high angular resolution γ-ray astronomy and polarimetry from the MeV to the TeV

    Energy Technology Data Exchange (ETDEWEB)

    Bernard, D., E-mail: denis.bernard@in2p3.fr [LLR, Ecole Polytechnique, CNRS/IN2P3, 91128 Palaiseau (France)

    2013-08-01

    We propose a “thin” detector as a high-angular-precision telescope and polarimeter for cosmic γ-rays above the pair-creation threshold. We have built a demonstrator based on a gaseous TPC. We are presently characterizing the detector with charged cosmic rays in the laboratory. Here we present some of its properties.

  18. Near-Intrinsic Energy Resolution for 30 to 662 keV Gamma Rays in a High Pressure Xenon Electroluminescent TPC

    CERN Document Server

    Álvarez, V; Cárcel, S; Castel, J; Cebrián, S; Cervera, A; Conde, C A N; Dafni, T; Dias, T H V T; Díaz, J; Egorov, M; Esteve, R; Evtoukhovitch, P; Fernandes, L M P; Ferrario, P; Ferreira, A L; Freitas, E D C; Gehman, V M; Gil, A; Goldschmidt, A; Gómez, H; Gómez-Cadenas, J J; González-Díaz, D; Gutiérrez, R M; Hauptman, J; Morata, J A Hernando; Herrera, D C; Iguaz, F J; Irastorza, I G; Jinete, M A; Labarga, L; Laing, A; Liubarsky, I; Lopes, J A M; Lorca, D; Losada, M; Luzón, G; Marí, A; Martín-Albo, J; Martínez, A; Miller, T; Moiseenko, A; Monrabal, F; Monteiro, C M B; Mora, F J; Moutinho, L M; Vidal, J Muñoz; da Luz, H Natal; Navarro, G; Nebot, M; Nygren, D; Oliveira, C A B; Palma, R; Pérez, J; Aparicio, J L Pérez; Renner, J; Ripoll, L; Rodríguez, A; Rodríguez, J; Santos, F P; Santos, J M F dos; Segui, L; Serra, L; Shuman, D; Simón, A; Sofka, C; Sorel, M; Toledo, J F; Tomás, A; Torrent, J; Tsamalaidze, Z; Vázquez, D; Veloso, J F C A; Villar, J A; Webb, R; White, J T; Yahlali, N

    2012-01-01

    We present the design, data and results from the NEXT prototype for Double Beta and Dark Matter (NEXT-DBDM) detector, a high-pressure gaseous natural xenon electroluminescent time projection chamber (TPC) that was built at the Lawrence Berkeley National Laboratory. It is a prototype of the planned NEXT-100 $^{136}$Xe neutrino-less double beta decay ($0\

  19. Measurement of negative particle multiplicity in S-Pb collisions at 200 GeV/c per nucleon with the NA36 TPC

    CERN Document Server

    Andersen, E; Brom, J M; Cherney, M; de la Cruz, B; Fernández, C; Garabatos, C; Garzón, J A; Geist, Walter M; Greiner, D E; Gruhn, Charles R; Hafidouni, M; Hrubec, Josef; Jones, P G; Judd, E G; Kuipers, J P M; Ladrem, M; Ladrón de Guevara, P; Løvhøiden, G; MacNaughton, J N; Mosquera, J; Natkaniec, Z; Nelson, J M; Neuhofer, Günther; Pérez de los Heros, C; Pló, M; Porth, Paul; Powell, B; Ramil, A; Rohringer, Herbert; Sakrejda, I; Thorsteinsen, T F; Traxler, J; Voltolini, C; Wozniak, K; Yañez, A; Zybert, R

    2001-01-01

    A high statistics study of the negative multiplicity distribution from S-Pb collisions at 200 GeV/c per nucleon is presented. The NA36 TPC was used to detect charged particles; corrections are based upon the maximum entropy method.

  20. Androgen receptor isoforms in human and rat prostate

    Institute of Scientific and Technical Information of China (English)

    Shu-JieXIA; Gang-YaoHAO; Xiao-DaTANG

    2000-01-01

    Aim: To investigate the androgen receptor (AR) isoforms and its variability of expression in human and rat prostatic tissues. Methods: Human benign prostatic hyperplasia (BPH) and prostatic cancer tissues were obtained from patients undergoing prostatectomy, and rat ventral prostate was incised 3 days after castration. Forty-one AR-positive BPH specimens, 3 prostatic cancer specimens, and 6 rat prostates were used. After processing at 4℃, the tissues were examined by means of high resolution isoelectric focusing (IEF) technique to determine their AR isoforms. Results:From the prostatic specimens, 3 types of AR isoforms were detected with pI values at 6.5, 6.0, and 5.3. In human BPH tissues, 15/41 (36.6%) specimens showed all the three types of isoforms, while 19/41 (46.3%) showed 2 isoforms at various combinations and 7/41(17.1%), 1 isoform. For the 3 prostatic cancer specimens, one showed 3 isoforms, one, 2 isoforms, and the other failed to show any isoform. All rat prostatic tissues showed 2 isoforms at different combinations. Binding of 3H-dihydrotestosterone (DHT) to the isoforms was inhibited by the addition of 100-fold excess of DHT or testosterone, but not progesterone, oestradiol or diethylstilboestrol. Conclusion: AR isoforms are different in different patients. Although their genesis is not clear, the therapeutic implication of the present observation appears to be interesting, that may help clarifying the individual differences in the response to hormonal therapy.(Asian J Androl 2000 Dec;2:307-310)

  1. Measurement of a 127 keV Neutron Field with the Future Standard Spectrometer micro-TPC

    CERN Document Server

    Maire, D; Bosson, G; Bourrion, O; Guillaudin, O; Lamblin, J; Lebreton, L; Mayet, F; Médard, J; Muraz, J F; Petit, M; Richer, J P; Riffard, Q; Santos, D

    2013-01-01

    In order to measure energy of neutron fields, with energy ranging from 8 keV to 1 MeV, a new primary standard is being developed at the IRSN (Institute for Radioprotection and Nuclear Safety). This project, micro-TPC (Micro Time Projection Chamber), carried out in collaboration with the LPSC (Laboratoire de Physique Subatomique et de Cosmologie), is based on the nucleus recoil detector principle. The instrument will be presented with the associated method to measure the neutron energy. This article will emphasize the proton energy calibration procedure and energy measurements of a neutron field produced at 127 keV on the IRSN facility AMANDE. Finally the COMIMAC device, dedicated to the calibration, will be described. This original device, developed at the LPSC, is able to produce proton and electron beams with an accurate energy ranging from 1 keV to 50 keV.

  2. Muon momentum measurement in ICARUS-T600 LAr-TPC via multiple scattering in few-GeV range

    Science.gov (United States)

    Antonello, M.; Baibussinov, B.; Bellini, V.; Benetti, P.; Boffelli, F.; Bubak, A.; Calligarich, E.; Centro, S.; Cervi, T.; Cesana, A.; Cieslik, K.; Cocco, A. G.; Dabrowska, A.; Dermenev, A.; Falcone, A.; Farnese, C.; Fava, A.; Ferrari, A.; Gibin, D.; Gninenko, S.; Guglielmi, A.; Haranczyk, M.; Holeczek, J.; Janik, M.; Kirsanov, M.; Kisiel, J.; Kochanek, I.; Lagoda, J.; Menegolli, A.; Meng, G.; Montanari, C.; Otwinowski, S.; Picchi, P.; Pietropaolo, F.; Plonski, P.; Rappoldi, A.; Raselli, G. L.; Rossella, M.; Rubbia, C.; Sala, P.; Scaramelli, A.; Sergiampietri, F.; Spanu, M.; Stefan, D.; Sulej, R.; Szarska, M.; Terrani, M.; Torti, M.; Tortorici, F.; Varanini, F.; Ventura, S.; Vignoli, C.; Wang, H.; Yang, X.; Zalewska, A.; Zani, A.; Zaremba, K.

    2017-04-01

    The measurement of muon momentum by Multiple Coulomb Scattering is a crucial ingredient to the reconstruction of νμ CC events in the ICARUS-T600 liquid argon TPC in absence of magnetic field, as in the search for sterile neutrinos at Fermilab where ICARUS will be exposed to ~ 1 GeV Booster neutrino beam. A sample of ~ 1000 stopping muons produced by charged current interactions of CNGS νμ in the surrounding rock at the INFN Gran Sasso underground Laboratory provides an ideal benchmark in the few-GeV range since their momentum can be directly and independently obtained by the calorimetric measurement. Stopping muon momentum in the 0.5-4.5 GeV/c range has been reconstructed via Multiple Coulomb Scattering with resolution ranging from 10 to 25% depending on muon energy, track length and uniformity of the electric field in the drift volume.

  3. Pattern recognition of $^{136}$Xe double beta decay events and background discrimination in a high pressure Xenon TPC

    CERN Document Server

    Cebrian, S; Gomez, H; Herrera, D C; Iguaz, F J; Irastorza, I G; Luzon, G; Segui, L; Tomas, A

    2013-01-01

    High pressure gas detectors offer advantages for the detection of rare events, where background reduction is crucial. For the neutrinoless double beta decay of 136Xe a high pressure xenon gas Time Projection Chamber (TPC) combines a good energy resolution and a detailed topological information of each event. The ionization topology of the double beta decay event of 136Xe in gaseous xenon has a characteristic shape defined by the two straggling electron tracks ending up in two higher ionization charge density blobs. With a properly pixelized readout, this topological information is invaluable to perform powerful background discrimination. In this study we carry out detailed simulations of the signal topology, as well as the competing topologies from gamma events that typically compose the background at these energies. We define observables based on graph theory concepts and develop automated discrimination algorithms which reduce the background level in around three orders of magnitude while keeping signal eff...

  4. Studies of THGEM-based detector at low-pressure Hydrogen/Deuterium, for AT-TPC applications

    CERN Document Server

    Cortesi, Marco; Mittig, Wolfgang; Bazin, Daniel; Beceiro-Novo, Saul; Stolz, Andreas

    2015-01-01

    We study the performance of single- and double- THick Gas Electron Multiplier (THGEM) detectors in pure Hydrogen and Deuterium at low pressures, in the range of 100-450 Torr. The effect of the pressure on the maximum achievable gain, ion-back flow and long-term gain stability are investigated for single and double cascade detectors. In particular, it was found that maximum achievable gains above 10^4, from single-photoelectrons avalanche, can be achieved for pressures of 200 Torr and above; for lower pressure the gains are limited by avalanche-induced secondary effects to a values of around 103. The results of this work are relevant in the field of avalanche mechanism in low-pressure, low-mass noble gases, in particular for applications of THGEM end-cap readout for active-target Time Projection Chambers (TPC) in the field of nuclear physics and nuclear astrophysics.

  5. The landscape of isoform switches in human cancers

    DEFF Research Database (Denmark)

    Vitting-Seerup, Kristoffer; Sandelin, Albin Gustav

    2017-01-01

    developed methods for identification and visualization of isoform switches with predicted functional consequences. Using these methods, we characterized isoform switching in RNA-seq data from >5,500 cancer patients covering 12 solid cancer types. Isoform switches with potential functional consequences were...... highly predictive of patient survival independent of cancer types. Our data constitute an important resource for cancer researchers, available through interactive web tools. Moreover, our methods, available as an R package, enable systematic analysis of isoform switches from other RNA-seq datasets...

  6. Membrane purifier prototype for hydrogen purification from towngas

    Energy Technology Data Exchange (ETDEWEB)

    Li, A.; Boyd, T.; Gulamhusein, A.; Grace, J.R.; Lim, C.J. [Membrane Reactor Technologies Ltd., Vancouver, BC (Canada)

    2007-07-01

    A prototype membrane purifier designed to purify hydrogen from towngas was described. The purifier was developed as a result of growing demands for metallic membrane purifiers. This paper provided details of a simulation conducted to observe the membrane's ability to purify towngas. The purifier was comprised of a cast-pressured vessel and a membrane module stack able to accommodate up to 6 palladium-silver (Pd-Ag) alloy membrane modules. Performance of the purifier was tested with both a hydrogen and a hydrogen and nitrogen oxide (N{sub 2}) mixture at 400 degrees C under various operating conditions. Results of the study showed that the purifier remained stable during temperature cycling. The hydrogen permeation rate followed Sievert's law in the tested temperature range. The hydrogen permeation rate increased when towngas flow rates and feeding pressures were increased. Hydrogen diffusion through the alloy membrane determined permeation rates through the membrane modules. No N{sub 2} leaks were detected using the mixtures. 1 ref., 1 tab., 6 figs.

  7. 75 FR 73035 - Purified Carboxymethylcellulose From Finland; Notice of Final Results of Antidumping Duty...

    Science.gov (United States)

    2010-11-29

    ... International Trade Administration Purified Carboxymethylcellulose From Finland; Notice of Final Results of... order on purified carboxymethylcellulose from Finland. See Purified Carboxymethylcellulose from Finland... order covering purified carboxymethylcellulose from Finland. See Preliminary Results. The...

  8. Functional refolding and characterization of two Tom40 isoforms from human mitochondria.

    Science.gov (United States)

    Mager, Frauke; Gessmann, Dennis; Nussberger, Stephan; Zeth, Kornelius

    2011-07-01

    Tom40 proteins represent an essential class of molecules which facilitate translocation of unfolded proteins from the cytosol into the mitochondrial intermembrane space. They are part of a high-molecular mass complex that forms the protein-conducting channel in outer mitochondrial membranes. This study concerns the recombinant expression, purification and folding of amino-terminally truncated variants of the two human Tom40 isoforms for structural biology experiments. Both CD and FTIR secondary structure analysis revealed a dominant beta-sheet structure and a short alpha-helical part for both proteins together with a high thermal stability. Two secondary structure elements can be denatured independently. Reconstitution of the recombinant protein into planar lipid bilayers demonstrated ion channel activity similar to Tom40 purified from Neurospora crassa mitochondrial membranes, but conductivity fingerprints differ from the structurally closely related VDAC proteins.

  9. Isoforms of Melanopsin Mediate Different Behavioral Responses to Light

    Science.gov (United States)

    Jagannath, Aarti; Hughes, Steven; Abdelgany, Amr; Pothecary, Carina A.; Di Pretoro, Simona; Pires, Susana S.; Vachtsevanos, Athanasios; Pilorz, Violetta; Brown, Laurence A.; Hossbach, Markus; MacLaren, Robert E.; Halford, Stephanie; Gatti, Silvia; Hankins, Mark W.; Wood, Matthew J.A.; Foster, Russell G.; Peirson, Stuart N.

    2015-01-01

    Summary Melanopsin (OPN4) is a retinal photopigment that mediates a wide range of non-image-forming (NIF) responses to light [1, 2] including circadian entrainment [3], sleep induction [4], the pupillary light response (PLR) [5], and negative masking of locomotor behavior (the acute suppression of activity in response to light) [6]. How these diverse NIF responses can all be mediated by a single photopigment has remained a mystery. We reasoned that the alternative splicing of melanopsin could provide the basis for functionally distinct photopigments arising from a single gene. The murine melanopsin gene is indeed alternatively spliced, producing two distinct isoforms, a short (OPN4S) and a long (OPN4L) isoform, which differ only in their C terminus tails [7]. Significantly, both isoforms form fully functional photopigments [7]. Here, we show that different isoforms of OPN4 mediate different behavioral responses to light. By using RNAi-mediated silencing of each isoform in vivo, we demonstrated that the short isoform (OPN4S) mediates light-induced pupillary constriction, the long isoform (OPN4L) regulates negative masking, and both isoforms contribute to phase-shifting circadian rhythms of locomotor behavior and light-mediated sleep induction. These findings demonstrate that splice variants of a single receptor gene can regulate strikingly different behaviors. PMID:26320947

  10. Tocopherol isoforms in parenteral lipid emulsions and neutrophil activation.

    NARCIS (Netherlands)

    Wanten, G.J.A.; Beunk, J.; Naber, A.H.J.; Swinkels, D.W.

    2002-01-01

    BACKGROUND AND AIMS: Tocopherol is a lipid-soluble anti-oxidant that exists in several isoforms. Patients on total parenteral nutrition depend on lipid emulsions for their tocopherol intake. In the present study, we analysed the content of tocopherol isoforms in various lipid emulsions. We also

  11. Direct interaction of the mouse cytomegalovirus m152/gp40 immunoevasin with RAE-1 isoforms.

    Science.gov (United States)

    Zhi, Li; Mans, Janet; Paskow, Michael J; Brown, Patrick H; Schuck, Peter; Jonjić, Stipan; Natarajan, Kannan; Margulies, David H

    2010-03-23

    Cytomegaloviruses (CMVs) are ubiquitous species-specific viruses that establish acute, persistent, and latent infections. Both human and mouse CMVs encode proteins that inhibit the activation of natural killer (NK) cells by downregulating cellular ligands for the NK cell activating receptor, NKG2D. The MCMV glycoprotein m152/gp40 downregulates the surface expression of RAE-1 to prevent NK cell control in vivo. So far, it is unclear if there is a direct interaction between m152 and RAE-1 and, if so, if m152 interacts differentially with the five identified RAE-1 isoforms, which are expressed as two groups in MCMV-susceptible or -resistant mouse strains. To address these questions, we expressed and purified the extracellular domains of RAE-1 and m152 and performed size exclusion chromatography binding assays as well as analytical ultracentrifugation and isothermal titration calorimetry to characterize these interactions quantitatively. We further evaluated the role of full-length and naturally glycosylated m152 and RAE-1 in cotransfected HEK293T cells. Our results confirmed that m152 binds RAE-1 directly, relatively tightly (K(d) RAE-1 isoforms, corresponding to the susceptibility to downregulation by m152. A PLWY motif found in RAE-1beta, although contributing to its affinity for m152, does not influence the affinity of RAE-1gamma or RAE-1delta, suggesting that other differences contribute to the RAE-1-m152 interaction. Molecular modeling of the different RAE-1 isoforms suggests a potential site for the m152 interaction.

  12. Evidence for leptin receptor isoforms heteromerization at the cell surface.

    Science.gov (United States)

    Bacart, Johan; Leloire, Audrey; Levoye, Angélique; Froguel, Philippe; Jockers, Ralf; Couturier, Cyril

    2010-06-01

    Leptin mediates its metabolic effects through several leptin receptor (LEP-R) isoforms. In humans, long (LEPRb) and short (LEPRa,c,d) isoforms are generated by alternative splicing. Most of leptin's effects are believed to be mediated by the OB-Rb isoform. However, the role of short LEPR isoforms and the possible existence of heteromers between different isoforms are poorly understood. Using BRET1 and optimized co-immunoprecipitation, we observed LEPRa/b and LEPRb/c heteromers located at the plasma membrane and stabilized by leptin. Given the widespread coexpression of LEPRa and LEPRb, our results suggest that LEPRa/b heteromers may represent a major receptor species in most tissues.

  13. Localization and functional characterization of the human NKCC2 isoforms

    DEFF Research Database (Denmark)

    Carota, I; Theilig, F; Oppermann, M;

    2010-01-01

    AIM: Salt reabsorption across the apical membrane of cells in the thick ascending limb (TAL) of Henle is primarily mediated by the bumetanide-sensitive Na(+)/K(+)/2Cl(-) cotransporter NKCC2. Three full-length splice variants of NKCC2 (NKCC2B, NKCC2A and NKCC2F) have been described. The NKCC2...... isoforms have specific localizations and transport characteristics, as assessed for rabbit, rat and mouse. In the present study, we aimed to address the localization and transport characteristics of the human NKCC2 isoforms. METHODS: RT-PCR, in situ hybridization and uptake studies in Xenopus oocytes were...... performed to characterize human NKCC2 isoforms. RESULTS: All three classical NKCC2 isoforms were detected in the human kidney; in addition, we found splice variants with tandem duplicates of the variable exon 4. Contrary to rodents, in which NKCC2F is the most abundant NKCC2 isoform, NKCC2A was the dominant...

  14. Development and characterization of human monoclonal antibodies that neutralize multiple TGFβ isoforms

    National Research Council Canada - National Science Library

    Bedinger, Daniel; Lao, Llewelyn; Khan, Shireen; Lee, Steve; Takeuchi, Toshihiko; Mirza, Amer M

    .... There are 3 main isoforms, TGFβ1, 2, and 3. As multiple TGFβ isoforms are involved in disease processes, maximal therapeutic efficacy may require neutralization of 2 or more of the TGFβ isoforms...

  15. 14-3-3 isoforms bind directly exon B of the 5'-UTR of human surfactant protein A2 mRNA.

    Science.gov (United States)

    Noutsios, Georgios T; Ghattas, Paul; Bennett, Stephanie; Floros, Joanna

    2015-07-15

    Human surfactant protein (SP) A (SP-A), an innate immunity molecule, is encoded by two genes, SFTPA1 and SFTPA2. The 5'-untranslated splice variant of SP-A2 (ABD), but not SP-A1 (AD), contains exon B (eB). eB is an enhancer for transcription and translation and contains cis-regulatory elements. Specific trans-acting factors, including 14-3-3, bind eB. The 14-3-3 protein family contains seven isoforms that have been found by mass spectrometry in eB electromobility shift assays (Noutsios et al. Am J Physiol Lung Cell Mol Physiol 304: L722-L735, 2013). We used four different approaches to investigate whether 14-3-3 isoforms bind directly to eB. 1) eB RNA pulldown assays showed that 14-3-3 isoforms specifically bind eB. 2) RNA electromobility shift assay complexes were formed using purified 14-3-3 isoforms β, γ, ε, η, σ, and τ, but not isoform ζ, with wild-type eB RNA. 3 and 4) RNA affinity chromatography assays and surface plasmon resonance analysis showed that 14-3-3 isoforms β, γ, ε, η, σ, and τ, but not isoform ζ, specifically and directly bind eB. Inhibition of 14-3-3 isoforms γ, ε, η, and τ/θ with shRNAs in NCI-H441 cells resulted in downregulation of SP-A2 levels but did not affect SP-A1 levels. However, inhibition of 14-3-3 isoform σ was correlated with lower levels of SP-A1 and SP-A2. Inhibition of 14-3-3 isoform ζ/δ, which does not bind eB, had no effect on expression levels of SP-A1 and SP-A2. In conclusion, the 14-3-3 protein family affects differential regulation of SP-A1 and SP-A2 by binding directly to SP-A2 5'-UTR mRNA.

  16. Study of gas purifiers for the CMS RPC detector

    CERN Document Server

    Benussi, L; Colafranceschi, S; Fabbri, F L; Felli, F; Ferrini, M; Giardoni, M; Greci, T; Paolozzi, A; Passamonti, L; Piccolo, D; Pierluigi, D; Russo, A; Saviano, G; Buontempo, S; Cimmino, A; de Gruttola, M; Fabozzi, F; Iorio, A O M; Lista, L; Paolucci, P; Baesso, P; Belli, G; Pagano, D; Ratti, S P; Vicini, A; Vitulo, P; Viviani, C; Guida, R; Sharma, A

    2012-01-01

    The CMS RPC muon detector utilizes a gas recirculation system called closed loop (CL) to cope with large gas mixture volumes and costs. A systematic study of CL gas purifiers has been carried out over 400 days between July 2008 and August 2009 at CERN in a low-radiation test area, with the use of RPC chambers with currents monitoring, and gas analysis sampling points. The study aimed to fully clarify the presence of pollutants, the chemistry of purifiers used in the CL, and the regeneration procedure. Preliminary results on contaminants release and purifier characterization are reported.

  17. Engineering of insulin receptor isoform-selective insulin analogues.

    Directory of Open Access Journals (Sweden)

    Tine Glendorf

    Full Text Available BACKGROUND: The insulin receptor (IR exists in two isoforms, A and B, and the isoform expression pattern is tissue-specific. The C-terminus of the insulin B chain is important for receptor binding and has been shown to contact the IR just adjacent to the region where the A and B isoforms differ. The aim of this study was to investigate the importance of the C-terminus of the B chain in IR isoform binding in order to explore the possibility of engineering tissue-specific/liver-specific insulin analogues. METHODOLOGY/PRINCIPAL FINDINGS: Insulin analogue libraries were constructed by total amino acid scanning mutagenesis. The relative binding affinities for the A and B isoform of the IR were determined by competition assays using scintillation proximity assay technology. Structural information was obtained by X-ray crystallography. Introduction of B25A or B25N mutations resulted in analogues with a 2-fold preference for the B compared to the A isoform, whereas the opposite was observed with a B25Y substitution. An acidic amino acid residue at position B27 caused an additional 2-fold selective increase in affinity for the receptor B isoform for analogues bearing a B25N mutation. Furthermore, the combination of B25H with either B27D or B27E also resulted in B isoform-preferential analogues (2-fold preference even though the corresponding single mutation analogues displayed no differences in relative isoform binding affinity. CONCLUSIONS/SIGNIFICANCE: We have discovered a new class of IR isoform-selective insulin analogues with 2-4-fold differences in relative binding affinities for either the A or the B isoform of the IR compared to human insulin. Our results demonstrate that a mutation at position B25 alone or in combination with a mutation at position B27 in the insulin molecule confers IR isoform selectivity. Isoform-preferential analogues may provide new opportunities for developing insulin analogues with improved clinical benefits.

  18. Immunodiffusion Studies of Purified Equine Infectious Anemia Virus

    Science.gov (United States)

    Nakajima, Hideo; Ushimi, Chuzo

    1971-01-01

    Antigenicity of purified equine infectious anemia (EIA) virus was examined by immunodiffusion against sera obtained from horses experimentally infected with EIA virus. The purified virus reacted with the infected horse serum, and virus-specific precipitating antibody was demonstrated. Furthermore, it was found that purified EIA virus reacted against the serum of horses infected with all strains of EIA virus which were antigenically different from one another. From the result, group-specific components of the virus rather than strain-specific ones were considered to be involved in the reaction. Serological reactivity was lost by adding antiserum from the infected horse to the antigen. The precipitating antibody usually appeared in the serum 1 to 2 weeks after the first febrile attack of EIA and remained for a longer period. Some characteristics of the purified antigen and specificity of the reaction for EIA are described. Images PMID:16557982

  19. Recent advances with a hybrid micro-pattern gas detector operated in low pressure H2 and He, for AT-TPC applications

    CERN Document Server

    Cortesi, Marco; Bazin, Daniel; Beceiro-Novo, Saul; Yurkon, John; Tanani, Rim Soussi; Wolff, Michael; Stolz, Andreas

    2015-01-01

    In view of a possible application as a charge-particle track readout for an Active-Target Time Projection Chamber (AT-TPC), the operational properties and performances of a hybrid Micro-Pattern Gaseous Detector (MPGD) were investigated in pure low-pressure Hydrogen (H2) and Helium (He). The detector consists of a MICROMesh GAseous Structure (MICROMEGAS) coupled to a single- or multi-cascade THick Gaseous Electron Multiplier (THGEM) as a pre-amplification stage. This study reports of the effective gain dependence of the hybrid-MPGD at relevant pressure (in the range of 200-760 torr) for different detector arrangements. The results of this work are relevant in the field of avalanche mechanism in low-pressure, low-mass noble gases, in particularly for applications of MPGD end-cap readout for active-target Time Projection Chambers (TPC) in the field of nuclear physics and nuclear astrophysics.

  20. Optimization of ultrasound-assisted extraction of total monomeric anthocyanin (TMA) and total phenolic content (TPC) from eggplant (Solanum melongena L.) peel.

    Science.gov (United States)

    Dranca, Florina; Oroian, Mircea

    2016-07-01

    The present study describes the extraction of total monomeric anthocyanin (TMA) and total phenolic content (TPC) from eggplant peel using ultrasonic treatments and methanol and 2-propanol as extraction solvents. The extraction yields were optimized by varying the solvent concentration, ultrasonic frequency, temperature and time of ultrasonic treatment. Box-Behnken design was used to investigate the effect of process variables on the ultrasound-assisted extraction. The results showed that for TPC extraction the optimal condition were obtained with a methanol concentration of 76.6%, 33.88 kHz ultrasonic frequency, a temperature of 69.4 °C and 57.5 min extraction time. For TMA the optimal condition were the following: 54.4% methanol concentration, 37 kHz, 55.1 °C and process time of 44.85 min. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Comparative study of the effects of solid-state fermentation with three filamentous fungi on the total phenolics content (TPC), flavonoids, and antioxidant activities of subfractions from oats (Avena sativa L.).

    Science.gov (United States)

    Cai, Shengbao; Wang, Ou; Wu, Wei; Zhu, Songjie; Zhou, Feng; Ji, Baoping; Gao, Fengyi; Zhang, Di; Liu, Jia; Cheng, Qian

    2012-01-11

    The aim of present work was to investigate the effect of solid-state fermentation with filamentous fungi (Aspergillus oryzae var. effuses, Aspergillus oryzae, and Aspergillus niger) on total phenolics content (TPC), flavonoids, and antioxidant activities of four subfractions of oat, namely, n-hexane, ethyl acetate (EA), n-butanol, and water, and compare them to their corresponding subfractions of unfermented oat. The TPC and total flavonoids increased dramatically, especially in EA subfractions (p < 0.05). The levels of antioxidant activity of subfractions were also significantly enhanced (p < 0.05). The highest antioxidant activities were also found in the EA subfractions. The polyphenols in EA were analyzed by high-performance liquid chromatography at 280 nm. Most polyphenols were increased remarkably, especially ferulic and caffeic acids. There was a clear correlation between the TPC and antioxidant activity. In conclusion, fungi fermentation is a potential bioprocess for increasing the TPC, flavonoids, and antioxidant activities of oat-based food.

  2. Adiponectin isoforms: a potential therapeutic target in rheumatoid arthritis?

    Science.gov (United States)

    Frommer, Klaus W; Schäffler, Andreas; Büchler, Christa; Steinmeyer, Jürgen; Rickert, Markus; Rehart, Stefan; Brentano, Fabia; Gay, Steffen; Müller-Ladner, Ulf; Neumann, Elena

    2012-10-01

    Several clinical studies have suggested the adipocytokine adiponectin is involved in the progression of rheumatoid arthritis (RA). From this point of view, adiponectin might present a new therapeutic target. However, as adiponectin also exerts beneficial effects in the human organism, a strategy that would allow its detrimental effects to be abolished while maintaining the positive effects would be highly favourable. To elucidate such a strategy, the authors analysed whether the different adiponectin isoforms induce diverging effects, especially with regard to rheumatoid arthritis synovial fibroblasts (RASF), a central cell type in RA pathogenesis capable of invading into and destroying cartilage. Affymetrix microarrays were used to screen for changes in gene expression of RASF. Messenger RNA levels were quantified by real-time PCR, protein levels by immunoassay. The migration of RASF and primary human lymphocytes was analysed using a two-chamber migration assay. In RASF, the individual adiponectin isoforms induced numerous genes/proteins relevant in RA pathogenesis to clearly different extents. In general, the most potent isoforms were the high molecular weight/middle molecular weight isoforms and the globular isoform, while the least potent isoform was the adiponectin trimer. The chemokines secreted by RASF upon adiponectin stimulation resulted in an increased migration of RASF and lymphocytes. The results clearly suggest a pro-inflammatory and joint-destructive role of all adiponectin isoforms in RA pathophysiology, indicating that in chronic inflammatory joint diseases the detrimental effects outweigh the beneficial effects of adiponectin.

  3. Study of gas purifiers in the CMS RPC detector

    CERN Document Server

    Saviano, Giovanna

    2010-01-01

    The CMS RPC muon detector utilizes a gas recirculation system (Closed Loop) to cope with high gas mixture volumes and costs. A systematic study of Closed Loop gas purifiers has been carried out in 2008 and 2009 at the ISR experimental area of CERN, with the use of RPC chambers with currents monitoring, and gas analysis sampling points. Results on contaminants release and purifier characterization are presented

  4. Nonmuscle myosin II isoforms coassemble in living cells.

    Science.gov (United States)

    Beach, Jordan R; Shao, Lin; Remmert, Kirsten; Li, Dong; Betzig, Eric; Hammer, John A

    2014-05-19

    Nonmuscle myosin II (NM II) powers myriad developmental and cellular processes, including embryogenesis, cell migration, and cytokinesis [1]. To exert its functions, monomers of NM II assemble into bipolar filaments that produce a contractile force on the actin cytoskeleton. Mammalian cells express up to three isoforms of NM II (NM IIA, IIB, and IIC), each of which possesses distinct biophysical properties and supports unique as well as redundant cellular functions [2-8]. Despite previous efforts [9-13], it remains unclear whether NM II isoforms assemble in living cells to produce mixed (heterotypic) bipolar filaments or whether filaments consist entirely of a single isoform (homotypic). We addressed this question using fluorescently tagged versions of NM IIA, IIB, and IIC, isoform-specific immunostaining of the endogenous proteins, and two-color total internal reflection fluorescence structured-illumination microscopy, or TIRF-SIM, to visualize individual myosin II bipolar filaments inside cells. We show that NM II isoforms coassemble into heterotypic filaments in a variety of settings, including various types of stress fibers, individual filaments throughout the cell, and the contractile ring. We also show that the differential distribution of NM IIA and NM IIB typically seen in confocal micrographs of well-polarized cells is reflected in the composition of individual bipolar filaments. Interestingly, this differential distribution is less pronounced in freshly spread cells, arguing for the existence of a sorting mechanism acting over time. Together, our work argues that individual NM II isoforms are potentially performing both isoform-specific and isoform-redundant functions while coassembled with other NM II isoforms.

  5. Cosmic ray tests of a GEM-based TPC prototype operated in Ar–CF{sub 4}–isobutane gas mixtures: II

    Energy Technology Data Exchange (ETDEWEB)

    Kobayashi, M., E-mail: makoto.kobayashi.exp@kek.jp [High Energy Accelerator Research Organization (KEK), Tsukuba 305-0801 (Japan); Yonamine, R. [Graduate University for Advanced Studies, KEK, Tsukuba 305-0801 (Japan); Tomioka, T. [Tokyo University of Agriculture and Technology, Koganei 184-8588 (Japan); Aoza, A. [Saga University, Saga 840-8502 (Japan); Bito, H. [Tokyo University of Agriculture and Technology, Koganei 184-8588 (Japan); Fujii, K. [High Energy Accelerator Research Organization (KEK), Tsukuba 305-0801 (Japan); Higashi, T. [Saga University, Saga 840-8502 (Japan); Hiramatsu, K. [Kinki University, Higashi-Osaka 577-8502 (Japan); Ikematsu, K. [High Energy Accelerator Research Organization (KEK), Tsukuba 305-0801 (Japan); Ishikawa, A. [Saga University, Saga 840-8502 (Japan); Kato, Y. [Kinki University, Higashi-Osaka 577-8502 (Japan); Kuroiwa, H. [Saga University, Saga 840-8502 (Japan); Matsuda, T. [High Energy Accelerator Research Organization (KEK), Tsukuba 305-0801 (Japan); Nitoh, O.; Ohta, H.; Sakai, K. [Tokyo University of Agriculture and Technology, Koganei 184-8588 (Japan); Settles, R.D. [Max Planck Institute for Physics, DE-80805 Munich (Germany); Sugiyama, A.; Tsuji, H. [Saga University, Saga 840-8502 (Japan); Watanabe, T. [Kogakuin University, Hachioji 192-0015 (Japan); and others

    2014-12-11

    The spatial resolution along the pad-row direction was measured with a GEM-based TPC prototype for the future linear collider experiment in order to understand its performance for tracks with finite projected angles with respect to the pad-row normal. The degradation of the resolution due to the angular pad effect was confirmed to be consistent with the prediction of a simple calculation taking into account the cluster-size distribution and the avalanche fluctuation.

  6. TREX-DM: a low-background Micromegas-based TPC for low-mass WIMP detection

    CERN Document Server

    Iguaz, F J; Aznar, F; Castel, J F; Cebrián, S; Dafni, T; García, J A; Irastorza, I G; Lagraba, A; Luzón, G; Peiró, A

    2015-01-01

    If Dark Matter is made of Weakly Interacting Massive Particles (WIMPs) with masses below 10-20 GeV, the corresponding nuclear recoils in mainstream WIMP experiments are of energies too close, or below, the experimental threshold. New detection techniques, focused on the use of light target nuclei together with low energy thresholds, are needed to get competitive sensitivity to the low-mass range of the WIMP parameter space. Gas Time Projection Chambers (TPCs) can be operated with a variety of target elements, and on account of the amplification in gas, very low thresholds are achievable. Recent advances in electronics and in novel radiopure TPC readouts, especially micro-mesh gas structure (Micromegas) are improving the scalability and low-background prospects of Micromegas-based TPCs. If we add their well-known tracking capabilities, they are a good detection option for the search of low-mass WIMPs. Here we present TREX-DM, a prototype built to test this concept. The detector is designed to host an active ma...

  7. Improved background rejection in neutrinoless double beta decay experiments using a magnetic field in a high pressure xenon TPC

    CERN Document Server

    Renner, J; Hernando, J A; Imzaylov, A; Monrabal, F; Muñoz, J; Nygren, D; Gomez-Cadenas, J J

    2015-01-01

    We demonstrate that the application of an external magnetic field could lead to an improved background rejection in neutrinoless double-beta (0nbb) decay experiments using a high pressure xenon (HPXe) TPC. HPXe chambers are capable of imaging electron tracks, a feature that enhances the separation between signal events (the two electrons emitted in the 0nbb decay of 136Xe) and background events, arising chiefly from single electrons of kinetic energy compatible with the end-point of the 0nbb decay (Qbb ). Applying an external magnetic field of sufficiently high intensity (in the range of 0.5-1 Tesla for operating pressures in the range of 5-15 atmospheres) causes the electrons to produce helical tracks. Assuming the tracks can be properly reconstructed, the sign (direction) of curvature can be determined at several points along these tracks, and such information can be used to separate signal (0nbb) events containing two electrons producing a track with two different directions of curvature from background (s...

  8. Expression of Ca²⁺-permeable two-pore channels rescues NAADP signalling in TPC-deficient cells.

    Science.gov (United States)

    Ruas, Margarida; Davis, Lianne C; Chen, Cheng-Chang; Morgan, Anthony J; Chuang, Kai-Ting; Walseth, Timothy F; Grimm, Christian; Garnham, Clive; Powell, Trevor; Platt, Nick; Platt, Frances M; Biel, Martin; Wahl-Schott, Christian; Parrington, John; Galione, Antony

    2015-07-02

    The second messenger NAADP triggers Ca(2+) release from endo-lysosomes. Although two-pore channels (TPCs) have been proposed to be regulated by NAADP, recent studies have challenged this. By generating the first mouse line with demonstrable absence of both Tpcn1 and Tpcn2 expression (Tpcn1/2(-/-)), we show that the loss of endogenous TPCs abolished NAADP-dependent Ca(2+) responses as assessed by single-cell Ca(2+) imaging or patch-clamp of single endo-lysosomes. In contrast, currents stimulated by PI(3,5)P2 were only partially dependent on TPCs. In Tpcn1/2(-/-) cells, NAADP sensitivity was restored by re-expressing wild-type TPCs, but not by mutant versions with impaired Ca(2+)-permeability, nor by TRPML1. Another mouse line formerly reported as TPC-null likely expresses truncated TPCs, but we now show that these truncated proteins still support NAADP-induced Ca(2+) release. High-affinity [(32)P]NAADP binding still occurs in Tpcn1/2(-/-) tissue, suggesting that NAADP regulation is conferred by an accessory protein. Altogether, our data establish TPCs as Ca(2+)-permeable channels indispensable for NAADP signalling.

  9. p53 Family: Role of Protein Isoforms in Human Cancer

    Directory of Open Access Journals (Sweden)

    Jinxiong Wei

    2012-01-01

    Full Text Available TP53, TP63, and TP73 genes comprise the p53 family. Each gene produces protein isoforms through multiple mechanisms including extensive alternative mRNA splicing. Accumulating evidence shows that these isoforms play a critical role in the regulation of many biological processes in normal cells. Their abnormal expression contributes to tumorigenesis and has a profound effect on tumor response to curative therapy. This paper is an overview of isoform diversity in the p53 family and its role in cancer.

  10. The potential of discrimination methods in a high pressure xenon TPC for the search of the neutrinoless double-beta decay of Xe-136

    CERN Document Server

    Iguaz, F J; Castel, J F; Cebrian, S; Dafni, T; Galan, J; Garza, J G; Irastorza, I G; Luzon, G; Mirallas, H; Ruiz-Choliz, E

    2016-01-01

    In the search for the neutrinoless double beta decay of $^{136}$Xe, a high pressure xenon time projection chamber (HPXe-TPC) has two advantages over liquid xenon TPCs: a better energy resolution and the access to topological features, which may provide extra discrimination from background events. The PandaX-III experiment has recently proposed a 200 kg HPXe-TPC based on Micromegas readout planes, to be located at the Jinping Underground Laboratory in China. Its detection concept is based on two results obtained within the T-REX project: Micromegas readouts can be built with extremely low levels of radioactivity; and the operation in xenon-trimethylamine at 10 bar in realistic experimental conditions has proven an energy resolution of 3% FWHM at the region of interest. In this work, two discrimination methods are applied to simulated signal and background data in a generic 200 kg HPXe-TPC, based on two well-known algorithms of graph theory: the identification of connections and the search for the longest path....

  11. The potential of discrimination methods in a high pressure xenon TPC for the search of the neutrinoless double-beta decay of Xe-136

    Science.gov (United States)

    Iguaz, F. J.; Aznar, F.; Castel, J. F.; Cebrián, S.; Dafni, T.; Galán, J.; Garza, J. G.; Irastorza, I. G.; Luzón, G.; Mirallas, H.; Ruiz-Choliz, E.

    2017-09-01

    In the search for the neutrinoless double beta decay of 136Xe, a high pressure xenon time projection chamber (HPXe-TPC) has two advantages over liquid xenon TPCs: a better energy resolution and the access to topological features, which may provide extra discrimination from background events. The PandaX-III experiment has recently proposed a 200 kg HPXe-TPC based on Micromegas readout planes, to be located at the Jinping Underground Laboratory in China. Its detection concept is based on two results obtained within the T-REX project: Micromegas readouts can be built with extremely low levels of radioactivity; and the operation in xenon-trimethylamine at 10 bar in realistic experimental conditions has proven an energy resolution of 3% FWHM at the region of interest. In this work, two discrimination methods are applied to simulated signal and background data in a generic 200 kg HPXe-TPC, based on two well-known algorithms of graph theory: the identification of connections and the search for the longest path. Rejection factors greater than 100 are obtained for small pixel sizes and a signal efficiency of 40%. Moreover, a new observable (the blob charge density) rejects better surface contaminations, which makes the use of a trigger signal (T 0) not imperative in this experiment.

  12. Targeted Proteomics Enables Simultaneous Quantification of Folate Receptor Isoforms and Potential Isoform-based Diagnosis in Breast Cancer.

    Science.gov (United States)

    Yang, Ting; Xu, Feifei; Fang, Danjun; Chen, Yun

    2015-11-17

    The distinct roles of protein isoforms in cancer are becoming increasingly evident. FRα and FRβ, two major isoforms of the folate receptor family, generally have different cellular distribution and tissue specificity. However, the presence of FRβ in breast tumors, where FRα is normally expressed, complicates this situation. Prior to applying any FR isoform-based diagnosis and therapeutics, it is essential to monitor the expression profile of FR isoforms in a more accurate manner. An LC-MS/MS-based targeted proteomics assay was developed and validated in this study because of the lack of suitable methodology for the simultaneous and specific measurement of highly homologous isoforms occurring at low concentrations. FRα and FRβ monitoring was achieved by measuring their surrogate isoform-specific peptides. Five human breast cell lines, isolated macrophages and 60 matched pairs of breast tissue samples were subjected to the analysis. The results indicated that FRβ was overexpressed in tumor-associated macrophages (TAMs) but not epithelial cells, in addition to an enhanced level of FRα in breast cancer cells and tissue samples. Moreover, the levels of the FR isoforms were evaluated according to the histology, histopathological features and molecular subtypes of breast cancer. Several positive associations with PR/ER and HER2 status and metastasis were revealed.

  13. Prediction Model of TPC Reception Iron Amount Based on PCA-GA-BP%基于PCA-GA-BP的TPC受铁量预测模型

    Institute of Scientific and Technical Information of China (English)

    黄颖雷; 刘业峰; 黄辉; 郑秉霖

    2009-01-01

    Prediction problem of torpedo ladle car (TPC) actual reception iron amount under the blast furnace is discussed,which has an important function to coordinate molten iron balance,decrease exchanging bottles,reduce temperature loss and raise TPC utilization.A prediction model of TPC reception iron amount based on PCA-GA-BP is proposed.The principle component analysis is used to select the most relevant process features and to eliminate the correlations of the input variables.Back-propagation neural network is used to characterize the nonlinearity and accuracy.Genetic algorithm is employed to optimize the parameters and structure of the BP neural network by improving GA fitness function.Experiment results through the actual production data of an enterprise show the prediction rationality and validity,and the prediction accuracy of TiC actual reception iron amount is increased.%预测鱼雷罐车(TPC)在高炉的实际受铁量,时协调铁水平衡、减少兑罐次数和温降损失,保证高炉出铁安全,提高TPC利用率具有重要作用.采用主成分分析(PCA)提取过程特征参数,并剔除相关冗余信息;BP神经网络用来逼近受铁量预测这一非线性过程;改进了遗传算法(GA)的适应度函数,并精确给定BP神经网络的权值和阈值,进而建立了基于PCA-GA-BP的TPC受铁量预测模型.采用某钢铁企业实际生产数据运算,结果表明模型合理、有效,提高了鱼雷罐车(TPC)受铁量预测准确性.

  14. Sera and conditioned media contain different isoforms of platelet-derived growth factor (PDGF) which bind to different classes of PDGF receptor.

    Science.gov (United States)

    Bowen-Pope, D F; Hart, C E; Seifert, R A

    1989-02-15

    Platelet-derived growth factor (PDGF) is encoded by separate genes for two possible subunit chains (A-chain and B-chain) which can form three possible dimers (AA, AB, and BB). We have recently presented evidence that multiple forms of PDGF receptor exist which distinguish between these isoforms (Hart, C. H., Forstrom, J. W., Kelley, J. D., Smith, R. A., Ross, R., Murray, M. J., and Bowen-Pope, D. F. (1988) Science 240, 1529-1531). We used this specificity to determine the amount of PDGF from different sources which is able to bind to each class of receptor and found that each source had a characteristic isoform composition. Levels of total PDGF activity in sera from different species ranged more than 15-fold, from less than 1 ng/ml in dog, chicken, pig, and calf, to greater than 13 ng/ml in mouse and human. Despite these differences in PDGF content, the total mitogenic activities of the sera were comparable indicating that the relative importance of PDGF as a serum mitogen may vary considerably between species. Analysis of the total PDGF into the amounts of each isoform revealed great differences in composition. PDGF-BB constitutes only about 15% of the total binding activity in human PDGF purified by the method of Raines and Ross (Raines, E. W., and Ross, R. (1982) J. Biol. Chem. 257, 5154-5160) but is the predominant isoform in whole blood serum from all other species. In contrast to serum, medium conditioned by cultured PDGF-secreting cell types contained no detectable PDGF-BB except in two cases: medium conditioned by vascular endothelial cells and by cells transformed by simian sarcoma virus. The existence of isoform-specific PDGF receptors and the large variation in PDGF isoform composition dependent upon source may provide an important mechanism through which the effects of PDGF can be targeted to different cell types and/or toward eliciting different cell responses.

  15. Expression and functional analysis of two osmotin (PR5) isoforms with differential antifungal activity from Piper colubrinum: prediction of structure-function relationship by bioinformatics approach.

    Science.gov (United States)

    Mani, Tomson; Sivakumar, K C; Manjula, S

    2012-11-01

    Osmotin, a pathogenesis-related antifungal protein, is relevant in induced plant immunity and belongs to the thaumatin-like group of proteins (TLPs). This article describes comparative structural and functional analysis of the two osmotin isoforms cloned from Phytophthora-resistant wild Piper colubrinum. The two isoforms differ mainly by an internal deletion of 50 amino acid residues which separates them into two size categories (16.4 kDa-PcOSM1 and 21.5 kDa-PcOSM2) with pI values 5.6 and 8.3, respectively. Recombinant proteins were expressed in E. coli and antifungal activity assays of the purified proteins demonstrated significant inhibitory activity of the larger osmotin isoform (PcOSM2) on Phytophthora capsici and Fusarium oxysporum, and a markedly reduced antifungal potential of the smaller isoform (PcOSM1). Homology modelling of the proteins indicated structural alterations in their three-dimensional architecture. Tertiary structure of PcOSM2 conformed to the known structure of osmotin, with domain I comprising of 12 β-sheets, an α-helical domain II and a domain III composed of 2 β-sheets. PcOSM1 (smaller isoform) exhibited a distorted, indistinguishable domain III and loss of 4 β-sheets in domain I. Interestingly, an interdomain acidic cleft between domains I and II, containing an optimally placed endoglucanase catalytic pair composed of Glu-Asp residues, which is characteristic of antifungal PR5 proteins, was present in both isoforms. It is well accepted that the presence of an acidic cleft correlates with antifungal activity due to the presence of endoglucanase catalytic property, and hence the present observation of significantly reduced antifungal capacity of PcOSM1 despite the presence of a strong acidic cleft, is suggestive of the possible roles played by other structural features like domain I or/and III, in deciding the antifungal potential of osmotin.

  16. Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes.

    Science.gov (United States)

    Boros, Eszter; Szabó, András; Zboray, Katalin; Héja, Dávid; Pál, Gábor; Sahin-Tóth, Miklós

    2017-02-17

    Chymotrypsin-like elastases (CELAs) are pancreatic serine proteinases that digest dietary proteins. CELAs are typically expressed in multiple isoforms that can vary among different species. The human pancreas does not express CELA1 but secretes two CELA3 isoforms, CELA3A and CELA3B. The reasons for the CELA3 duplication and the substrate preferences of the duplicated isoforms are unclear. Here, we tested whether CELA3A and CELA3B evolved unique substrate specificities to compensate for the loss of CELA1. We constructed a phage library displaying variants of the substrate-like Schistocerca gregaria proteinase inhibitor 2 (SGPI-2) to select reversible high affinity inhibitors of human CELA3A, CELA3B, and porcine CELA1. Based on the reactive loop sequences of the phage display-selected inhibitors, we recombinantly expressed and purified 12 SGPI-2 variants and determined their binding affinities. We found that the primary specificity of CELA3A, CELA3B, and CELA1 was similar; all preferred aliphatic side chains at the so-called P1 position, the amino acid residue located directly N-terminal to the scissile peptide bond. P1 Met was an interesting exception that was preferred by CELA1 but weakly recognized by the CELA3 isoforms. The extended substrate specificity of CELA3A and CELA3B was comparable, whereas CELA1 exhibited unique interactions at several subsites. These observations indicated that the CELA1 and CELA3 paralogs have some different but also overlapping specificities and that the duplicated CELA3A and CELA3B isoforms did not evolve distinct substrate preferences. Thus, increased gene dosage rather than specificity divergence of the CELA3 isoforms may compensate for the loss of CELA1 digestive activity in the human pancreas.

  17. Isoform-specific targeting of ROCK proteins in immune cells

    OpenAIRE

    Zanin-Zhorov, Alexandra; Flynn, Ryan; Waksal, Samuel D.; Blazar, Bruce R.

    2016-01-01

    ABSTRACT Rho-associated kinase 1 (ROCK1) and ROCK2 are activated by Rho GTPase and control cytoskeleton rearrangement through modulating the phosphorylation of their down-stream effector molecules. Although these 2 isoforms share more than 90% homology within their kinase domain the question of whether ROCK proteins function identically in different cell types is not clear. By using both pharmacological inhibition and genetic knockdown approaches recent studies suggest that the ROCK2 isoform ...

  18. Neuronal profilin isoforms are addressed by different signalling pathways.

    Directory of Open Access Journals (Sweden)

    Kai Murk

    Full Text Available Profilins are prominent regulators of actin dynamics. While most mammalian cells express only one profilin, two isoforms, PFN1 and PFN2a are present in the CNS. To challenge the hypothesis that the expression of two profilin isoforms is linked to the complex shape of neurons and to the activity-dependent structural plasticity, we analysed how PFN1 and PFN2a respond to changes of neuronal activity. Simultaneous labelling of rodent embryonic neurons with isoform-specific monoclonal antibodies revealed both isoforms in the same synapse. Immunoelectron microscopy on brain sections demonstrated both profilins in synapses of the mature rodent cortex, hippocampus and cerebellum. Both isoforms were significantly more abundant in postsynaptic than in presynaptic structures. Immunofluorescence showed PFN2a associated with gephyrin clusters of the postsynaptic active zone in inhibitory synapses of embryonic neurons. When cultures were stimulated in order to change their activity level, active synapses that were identified by the uptake of synaptotagmin antibodies, displayed significantly higher amounts of both isoforms than non-stimulated controls. Specific inhibition of NMDA receptors by the antagonist APV in cultured rat hippocampal neurons resulted in a decrease of PFN2a but left PFN1 unaffected. Stimulation by the brain derived neurotrophic factor (BDNF, on the other hand, led to a significant increase in both synaptic PFN1 and PFN2a. Analogous results were obtained for neuronal nuclei: both isoforms were localized in the same nucleus, and their levels rose significantly in response to KCl stimulation, whereas BDNF caused here a higher increase in PFN1 than in PFN2a. Our results strongly support the notion of an isoform specific role for profilins as regulators of actin dynamics in different signalling pathways, in excitatory as well as in inhibitory synapses. Furthermore, they suggest a functional role for both profilins in neuronal nuclei.

  19. Cell-specific expression of TLR9 isoforms in inflammation.

    Science.gov (United States)

    McKelvey, Kelly J; Highton, John; Hessian, Paul A

    2011-02-01

    Toll-like receptors (TLRs) are key pattern recognition receptors during an immune response. With five isoforms of human TLR9 described, we hypothesised that differential expression of TLR9 isoforms in different cell types would result in variable contributions to the overall input from TLR9 during inflammation. We assessed the molecular expression of the TLR9 isoforms, TLR9-A, -C and -D. In normal peripheral blood mononuclear cells, B-lymphocytes express ∼100-fold more TLR9-A transcript than monocytes or T-lymphocytes, which predominantly express the TLR9-C transcript. Switches in isoform predominance accompany B-lymphocyte development. TLR9 protein expression in rheumatoid inflammatory lesions reflected the TLR9 isoform expression by immune cells. Herein we suggest that B-lymphocytes and plasmacytoid dendritic cells contribute the ∼3-fold higher TLR9-A transcript levels observed in inflamed synovium when compared to subcutaneous rheumatoid nodules. In contrast, macrophages and T-lymphocytes contribute the ∼4-fold higher TLR9-C transcript levels seen in nodules, compared to synovia. From protein sequence, predictions of subcellular localisation suggest TLR9-B may locate to the mitochondria, whereas TLR9-D adopts an opposing orientation in the endoplasmic reticulum. Consistent with this, structure models raise the possibility of alternative ligands for the TLR9-B and TLR9-D variants. Our results highlight differences in the expression of human TLR9 isoforms in normal and inflamed tissues, with differing contributions to inflammation.

  20. Yeast ADP/ATP Carrier Isoform 2

    Science.gov (United States)

    Clémençon, Benjamin; Rey, Martial; Trézéguet, Véronique; Forest, Eric; Pelosi, Ludovic

    2011-01-01

    The mitochondrial ADP/ATP carrier, or Ancp, is a member of the mitochondrial carrier family responsible for exchanging ADP and ATP across the mitochondrial inner membrane. ADP/ATP transport involves Ancp switching between two conformational states. These can be analyzed using specific inhibitors, carboxyatractyloside (CATR) and bongkrekic acid (BA). The high resolution three-dimensional structure of bovine Anc1p (bAnc1p), as a CATR-carrier complex, has been solved. However, because the structure of the BA-carrier complex has not yet been determined, the detailed mechanism of transport remains unknown. Recently, sample processing for hydrogen/deuterium exchange experiments coupled to mass spectrometry was improved, providing novel insights into bAnc1p conformational transitions due to inhibitor binding. In this work we performed both hydrogen/deuterium exchange-mass spectrometry experiments and genetic manipulations. Because these are very difficult to apply with bovine Anc1p, we used Saccharomyces cerevisiae Anc isoform 2 (ScAnc2p). Significant differences in solvent accessibility were observed throughout the amino acid sequence for ScAnc2p complexed to either CATR or BA. Interestingly, in detergent solution, the conformational dynamics of ScAnc2p were dissimilar to those of bAnc1p, in particular for the upper half of the cavity, toward the intermembrane space, and the m2 loop, which is thought to be easily accessible to the solvent from the matrix in bAnc1p. Our study then focused on the methionyl residues of the Ancp signature sequence, RRRMMM. All our results indicate that the methionine cluster is involved in the ADP/ATP transport mechanism and confirm that the Ancp cavity is a highly dynamic structure. PMID:21868387

  1. Recombinant erythropoietin in humans has a prolonged effect on circulating erythropoietin isoform distribution

    DEFF Research Database (Denmark)

    Aachmann-Andersen, Niels Jacob; Just Christensen, Søren; Lisbjerg, Kristian

    2014-01-01

    The membrane-assisted isoform immunoassay (MAIIA) quantitates erythropoietin (EPO) isoforms as percentages of migrated isoforms (PMI). We evaluated the effect of recombinant human EPO (rhEPO) on the distribution of EPO isoforms in plasma in a randomized, placebo-controlled, double-blinded, cross...

  2. Structural diversity and evolution of the N-terminal isoform-specific region of ecdysone receptor-A and -B1 isoforms in insects

    Directory of Open Access Journals (Sweden)

    Kubo Takeo

    2010-02-01

    Full Text Available Abstract Background The ecdysone receptor (EcR regulates various cellular responses to ecdysteroids during insect development. Insects have multiple EcR isoforms with different N-terminal A/B domains that contain the isoform-specific activation function (AF-1 region. Although distinct physiologic functions of the EcR isoforms have been characterized in higher holometabolous insects, they remain unclear in basal direct-developing insects, in which only A isoform has been identified. To examine the structural basis of the EcR isoform-specific AF-1 regions, we performed a comprehensive structural comparison of the isoform-specific region of the EcR-A and -B1 isoforms in insects. Results The EcR isoforms were newly identified in 51 species of insects and non-insect arthropods, including direct-developing ametabolous and hemimetabolous insects. The comprehensive structural comparison revealed that the isoform-specific region of each EcR isoform contained evolutionally conserved microdomain structures and insect subgroup-specific structural modifications. The A isoform-specific region generally contained four conserved microdomains, including the SUMOylation motif and the nuclear localization signal, whereas the B1 isoform-specific region contained three conserved microdomains, including an acidic activator domain-like motif. In addition, the EcR-B1 isoform of holometabolous insects had a novel microdomain at the N-terminal end. Conclusions Given that the nuclear receptor AF-1 is involved in cofactor recruitment and transcriptional regulation, the microdomain structures identified in the isoform-specific A/B domains might function as signature motifs and/or as targets for cofactor proteins that play essential roles in the EcR isoform-specific AF-1 regions. Moreover, the novel microdomain in the isoform-specific region of the holometabolous insect EcR-B1 isoform suggests that the holometabolous insect EcR-B1 acquired additional transcriptional

  3. Precision meson spectroscopy. Diffractive production at COMPASS and development of a GEM-based TPC for PANDA

    Energy Technology Data Exchange (ETDEWEB)

    Weitzel, Quirin

    2008-09-24

    . Due to its excellent tracking capabilities for charged particles, a time projection chamber (TPC) has been proposed for the central tracker of PANDA. A continuous operation without ion gate is foreseen, which constitutes a novel development in high-rate particle physics experiments. Gas Electron Multiplier (GEM) foils offer an intrinsic ion back-flow suppression combined with high gains, and will therefore be used for gas amplification. A small-size GEM-TPC test chamber has been constructed during this thesis and commissioned using both X-rays and muons from cosmic-ray air-showers. From the latter data, a spatial resolution down to 140 {mu}m has been achieved. The detector has been operated stably for many months with Ar/CO{sub 2} (70/30) and at typical gas amplification factors of (5-10).10{sup 3}. (orig.)

  4. Strong purifying selection at genes escaping X chromosome inactivation.

    Science.gov (United States)

    Park, Chungoo; Carrel, Laura; Makova, Kateryna D

    2010-11-01

    To achieve dosage balance of X-linked genes between mammalian males and females, one female X chromosome becomes inactivated. However, approximately 15% of genes on this inactivated chromosome escape X chromosome inactivation (XCI). Here, using a chromosome-wide analysis of primate X-linked orthologs, we test a hypothesis that such genes evolve under a unique selective pressure. We find that escape genes are subject to stronger purifying selection than inactivated genes and that positive selection does not significantly affect the evolution of these genes. The strength of selection does not differ between escape genes with similar versus different expression levels in males versus females. Intriguingly, escape genes possessing Y homologs evolve under the strongest purifying selection. We also found evidence of stronger conservation in gene expression levels in escape than inactivated genes. We hypothesize that divergence in function and expression between X and Y gametologs is driving such strong purifying selection for escape genes.

  5. Multiple isoform recovery (MIR)-PCR: a simple method for the isolation of related mRNA isoforms.

    OpenAIRE

    Fagotti, A; Gabbiani, G.; Pascolini, R; Neuville, P

    1998-01-01

    We present a rapid and efficient method for the detection of related transcripts with different expression levels. This approach combines the rapid amplification of cDNA ends (RACE) method with a cDNA subtractive technique. The strategy is based on successive subtractions of prevalent isoforms resulting in enrichment of less expressed transcripts. For each subtraction, a biotinylated primer specific for the prevalent isoform is hybridized on the total cDNA and the hybrid is retained on a stre...

  6. 76 FR 29194 - Purified Carboxymethylcellulose From Mexico and Sweden: Revocation of Antidumping Duty Orders

    Science.gov (United States)

    2011-05-20

    ... International Trade Administration Purified Carboxymethylcellulose From Mexico and Sweden: Revocation of... duty orders on purified carboxymethylcellulose from Mexico and Sweden. Pursuant to section 751(c) of... of the existing antidumping duty orders on purified carboxymethylcellulose from Mexico and...

  7. 75 FR 57815 - Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden

    Science.gov (United States)

    2010-09-22

    ... COMMISSION Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden AGENCY: United... antidumping duty orders on purified carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden... antidumping duty orders on purified carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden...

  8. 76 FR 29191 - Purified Carboxymethylcellulose From Finland and the Netherlands: Continuation of Antidumping...

    Science.gov (United States)

    2011-05-20

    ... International Trade Administration Purified Carboxymethylcellulose From Finland and the Netherlands... from Finland and the Netherlands would likely lead to continuation or recurrence of dumping and... orders on purified carboxymethylcellulose (purified CMC) from Finland and the Netherlands. See Notice...

  9. Distinct functional interactions between actin isoforms and nonsarcomeric myosins.

    Directory of Open Access Journals (Sweden)

    Mirco Müller

    Full Text Available Despite their near sequence identity, actin isoforms cannot completely replace each other in vivo and show marked differences in their tissue-specific and subcellular localization. Little is known about isoform-specific differences in their interactions with myosin motors and other actin-binding proteins. Mammalian cytoplasmic β- and γ-actin interact with nonsarcomeric conventional myosins such as the members of the nonmuscle myosin-2 family and myosin-7A. These interactions support a wide range of cellular processes including cytokinesis, maintenance of cell polarity, cell adhesion, migration, and mechano-electrical transduction. To elucidate differences in the ability of isoactins to bind and stimulate the enzymatic activity of individual myosin isoforms, we characterized the interactions of human skeletal muscle α-actin, cytoplasmic β-actin, and cytoplasmic γ-actin with human myosin-7A and nonmuscle myosins-2A, -2B and -2C1. In the case of nonmuscle myosins-2A and -2B, the interaction with either cytoplasmic actin isoform results in 4-fold greater stimulation of myosin ATPase activity than was observed in the presence of α-skeletal muscle actin. Nonmuscle myosin-2C1 is most potently activated by β-actin and myosin-7A by γ-actin. Our results indicate that β- and γ-actin isoforms contribute to the modulation of nonmuscle myosin-2 and myosin-7A activity and thereby to the spatial and temporal regulation of cytoskeletal dynamics. FRET-based analyses show efficient copolymerization abilities for the actin isoforms in vitro. Experiments with hybrid actin filaments show that the extent of actomyosin coupling efficiency can be regulated by the isoform composition of actin filaments.

  10. Differential activities of glucocorticoid-induced leucine zipper protein isoforms.

    Science.gov (United States)

    Soundararajan, Rama; Wang, Jian; Melters, Daniël; Pearce, David

    2007-12-14

    Glucocorticoid-induced leucine zipper protein (GILZ) is expressed in both epithelial and immune tissues and modulates a variety of cellular functions, including proliferation and epithelial sodium channel (ENaC) activity. A number of reports have described various GILZ activities, focusing on a single isoform with molecular mass of approximately 17 kDa, now termed GILZ1. In GILZ immunoblots using a newly developed antiserum, we detected multiple species in extracts from cultured kidney cells. Mass spectrometric analysis revealed that one of these represented a previously uncharacterized distinct isoform of GILZ, GILZ2. Rapid amplification of cDNA ends was used to clone cDNAs corresponding to four isoforms, which, in addition to GILZ1 and GILZ2, included new isoforms GILZ3 and GILZ4. Heterologous expression of these four GILZ isoforms in cultured cells revealed striking functional differences. Notably, GILZ1 was the only isoform that significantly stimulated ENaC-mediated Na+ current in a kidney collecting duct cell line, although GILZ2 and GILZ3 also stimulated ENaC surface expression in HEK 293 cells. GILZ1 and GILZ3, and to a lesser extent GILZ2, inhibited ERK phosphorylation. Interestingly, GILZ4, which had no effect on either ENaC or ERK, potently suppressed cellular proliferation, as did GILZ1, but not GILZ2 or GILZ3. Finally, rat and mouse tissues all expressed multiple GILZ species but varied in the relative abundance of each. These data suggest that multiple GILZ isoforms are expressed in most cells and tissues and that these play distinct roles in regulating key cellular functions, including proliferation and ion transport. Furthermore, GILZ inhibition of ERK appears to play an essential role in stimulation of cell surface ENaC but not in inhibition of proliferation.

  11. Purified guar galactomannan as an improved pharmaceutical excipient.

    Science.gov (United States)

    Gebert, M S; Friend, D R

    1998-08-01

    The purpose of this study was to assess certain pharmaceutical attributes of guar galactomannan, a hydrocolloid polysaccharide obtained from the endosperm of the leguminous plant Cyamopsis tetragonolobus (L.), following purification using both literature procedures and new processes. Experiments were performed to measure viscosity, hydration rate, tablet hardness, and dissolution profiles of guar galactomannan both before and after purification. The viscosity of an aqueous 1% purified galactomannan solution is typically 40-50% higher than its unpurified guar galactomannan precursor. The hydration rate of an aqueous 1% purified galactomannan solution increases by 100% after purification. These physicochemical changes resulted in improvements in pharmaceutical properties such as better stir speed independence in both tablet and capsule dissolution profiles and improved tablet hardness. For instance, time to 50% dissolution of ranitidine HCl from capsules containing unpurified guar gum was 0.4 and 1.8 hr at 20 and 40 rpm, respectively, using USP Apparatus II. Using the same amount of purified guar gum and the same conditions (20 and 40 rpm), these values were increased to 2.9 and 3.8 hr, respectively. These data demonstrate a reduced effect of changing agitation conditions and the need for less guar gum to sustain the release of a water-soluble drug. Tablet hardness of purified guar gum (particle size < 75 microns) was about 7 kP and the same unpurified guar gum of equal particle size and hydration gave a hardness of less than 1 kP.

  12. Partial characterization of hog renin purified by affinity chromatography.

    Science.gov (United States)

    Devaux, C; Ménard, J; Sicard, P; Corvol, P

    1976-05-01

    A method has been set up to purify renin on a large scale by affinity chromatography using Pepstatin, a potent inhibitor of renin, as a ligand. Pepstatin was covalently coupled to Sepharose via six different spacer 'arms'. The Sepharose-hexamethylenediamino-Pepstatin appeared to be the better derivative for renin purification even at a concentration as low as 160 nmol of Pepstatin/ml of moist gel. Renin was extracted from 100 kg of hog kidneys and semi-purified by ammonium sulfate precipitations and chromatography on DEAE-cellulose. The active fraction (48.5 g of proteins) was applied on a 500-ml affinity column. Renin was eluted in the starting buffer containing 6 M urea. Renin was purified 120-fold by the affinity chromatography step with a 79% recovery. Physico-chemical characterization of highly purified renin was performed. Isoelectrofocusing on a pH gradient from 3 to 6 showed a major peak with an isoelectric point (pI) of 4.95 and a minor peak (pI = 4.70). Polyacrylamide gel electrophoresis, pH 7.8, at different gel concentrations, showed a single peak of renin activity which was found in the major protein band. Molecular size estimated on agarose-acrylamide gel filtration was 40 000. All these physical parameters were similar before and after purification.

  13. Influence of a highly purified senna extract on colonic epithelium

    NARCIS (Netherlands)

    van Gorkom, B A; Karrenbeld, A; van Der Sluis, T; Koudstaal, J; de Vries, E G; Kleibeuker, J H

    2000-01-01

    BACKGROUND: Chronic use of sennoside laxatives often causes pseudomelanosis coli. A recent study suggested that pseudomelanosis coli is associated with an increased colorectal cancer risk. A single high dose of highly purified senna extract increased proliferation rate and reduced crypt length in th

  14. Mechanical Properties, Purifying Techniques and Processing Methods of Metal Yttrium

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The mechanical properties of metal yttrium such as strength, plasticity, hardness and elasticity were introduced. The purifying techniques of yttrium were discussed in detail. The processing methods for metal yttrium including extruding, forging, rolling, wiredrawing and welding were also introduced. Finally, the potential use of yttrium and its alloys were prospected.

  15. Strong Purifying Selection in Transmission of Mammalian Mitochondrial DNA

    Science.gov (United States)

    Stewart, James Bruce; Freyer, Christoph; Elson, Joanna L; Wredenberg, Anna; Cansu, Zekiye; Trifunovic, Aleksandra; Larsson, Nils-Göran

    2008-01-01

    There is an intense debate concerning whether selection or demographics has been most important in shaping the sequence variation observed in modern human mitochondrial DNA (mtDNA). Purifying selection is thought to be important in shaping mtDNA sequence evolution, but the strength of this selection has been debated, mainly due to the threshold effect of pathogenic mtDNA mutations and an observed excess of new mtDNA mutations in human population data. We experimentally addressed this issue by studying the maternal transmission of random mtDNA mutations in mtDNA mutator mice expressing a proofreading-deficient mitochondrial DNA polymerase. We report a rapid and strong elimination of nonsynonymous changes in protein-coding genes; the hallmark of purifying selection. There are striking similarities between the mutational patterns in our experimental mouse system and human mtDNA polymorphisms. These data show strong purifying selection against mutations within mtDNA protein-coding genes. To our knowledge, our study presents the first direct experimental observations of the fate of random mtDNA mutations in the mammalian germ line and demonstrates the importance of purifying selection in shaping mitochondrial sequence diversity. PMID:18232733

  16. Highly thermostable xylanase purified from Rhizomucor miehei NRL 3169.

    Science.gov (United States)

    Fawzi, E M

    2011-03-01

    A thermostable xylanase was purified and characterized from the thermophilic fungus Rhizomucor miehei (Cooney & Emerson) Schipper. The enzyme was purified to homogeneity by ammonium sulfate precipitation, sephadex G-100 gel filtration and diethylaminoethyl cellulose anion exchange chromatography with a 29.1-fold. The enzyme was highly active within a range of pH from 5.0 to 6.5. The optimum temperature of the purified enzyme was 75°C. The enzyme showed high thermal stability at 70°C and 75°C and the half-life of the xylanase at 90°C was 30 min. Km and Vmax values at 50°C of the purified enzyme were 0.055 mg/ml and 113.5 μmol min⁻¹ mg⁻¹ respectively. The enzyme was activated by Ca²+, Cu²+, K+ and Na+. On the other hand, Ag²+, Hg²+, Ba²+, and Zn²+ inhibited the enzyme. The molecular weight of the xylanase was estimated to be 27 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The present study is among the first works to examine and describe a secreted highly thermostable endoxylanase from the Rhizomucor miehei fungus. This enzyme displays a number of biochemical properties that make it a potentially strong candidate for industrial and commercial application in pulp bleaching.

  17. Inhibition of adenylyl and guanylyl cyclase isoforms by the antiviral drug foscarnet.

    Science.gov (United States)

    Kudlacek, O; Mitterauer, T; Nanoff, C; Hohenegger, M; Tang, W J; Freissmuth, M; Kleuss, C

    2001-02-02

    The pyrophosphate (PP(i)) analog foscarnet inhibits viral DNA-polymerases and is used to treat cytomegalovirus and human immunodeficiency vius infections. Nucleotide cyclases and DNA-polymerases catalyze analogous reactions, i.e. a phosphodiester bond formation, and have similar topologies in their active sites. Inhibition by foscarnet of adenylyl cyclase isoforms was therefore tested with (i) purified catalytic domains C1 and C2 of types I and VII (IC1 and VIIC1) and of type II (IIC2) and (ii) membrane-bound holoenzymes (from mammalian tissues and types I, II, and V heterologously expressed in Sf9 cell membranes). Foscarnet was more potent than PP(i) in suppressing forskolin-stimulated catalysis by both, IC1/IIC2 and VIIC1/IIC2. Stimulation of VIIC1/IIC2 by Galpha(s) relieved the inhibition by foscarnet but not that by PP(i). The IC(50) of foscarnet on membrane-bound adenylyl cyclases also depended on their mode of regulation. These findings predict that receptor-dependent cAMP formation is sensitive to inhibition by foscarnet in some, but not all, cells. This was verified with two cell lines; foscarnet blocked cAMP accumulation after A(2A)-adenosine receptor stimulation in PC12 but not in HEK-A(2A) cells. Foscarnet also inhibited soluble and, to a lesser extent, particulate guanylyl cylase. Thus, foscarnet interferes with the generation of cyclic nucleotides, an effect which may give rise to clinical side effects. The extent of inhibition varies with the enzyme isoform and with the regulatory input.

  18. Microbiological stability of homeopathic medicines using purified water as vehicle

    Directory of Open Access Journals (Sweden)

    Virginia Teresa Cegalla

    2011-09-01

    Full Text Available Background: Homeopathic medicines are prepared in homeopathic pharmacies. This leads to freedom of prescription but requires more knowledge of the clinicians to achieve the best results. Preparations made of purified water receive a validity of 24 hours, but there are prescriptions for up to 30 days. This contradiction raises tensions among physicians, pharmacists and patients. Aims: to evaluate the increase in microbiological contamination in homeopathic medicines using purified water as vehicle compared with the microbiological stability of purified water. Contribute to the quality of homeopathic medicine and treatment. Methodology: daily microbiological analysis for one week to assess the growth of heterotrophic bacteria, Pseudomonas, yeasts and molds. The reference used was the USP 32/NF 27 and the Brazilian Pharmacopoeia 5th edition. Results: there was a higher growth of microorganisms on the medicine, compared with purified water. From the 2nd day on, this growth has been beyond the legal limits. Discussion: medicines for oral use are not sterile preparations, but they must remain stable during its shelf life. Our results indicate that contamination occurs from the earliest days of use. This shows the need to change the prescription in relation of the vehicle, to ensure hygiene and avoid potential contamination of the patient. It is necessary to prevent conflict of information between pharmacists and patients, and the contradiction of the doctor's advice, besides the potential risk of responsibility to be attributed to the pharmacy. It is necessary to promote a discussion between pharmacists and clinicians, to spread this information for those that prescribe. Conclusion: there was an increased of microbiological contamination of the medicines dispensed in purified water, which harms the quality of homeopathic medicine and homeopathic treatment.

  19. Chemical origins of isoform selectivity in histone deacetylase inhibitors.

    Science.gov (United States)

    Butler, Kyle V; Kozikowski, Alan P

    2008-01-01

    Histones undergo extensive posttranslational modifications that affect gene expression. Acetylation is a key histone modification that is primarily regulated by two enzymes, one of which is histone deacetylase (HDAC). The activity of HDAC causes transcriptional silencing of DNA. Eleven distinct zinc-dependent histone deacetylase isoforms have been identified in humans. Each isoform has a unique structure and function, and regulates a unique set of genes. HDAC is responsible for the regulation of many genes involved in cancer cell proliferation, and it has been implicated in the pathogenesis of many neurological conditions. HDAC inhibitors are known to be very effective anti-cancer agents, and research has shown them to be potential treatments for many other conditions. Histone deacetylase inhibitors modify the expression of many genes, and it is possible that inhibition of one isoform could cause epigenetic changes that are beneficial to treatment of a disease, while inhibition of another isoform could cause contradictory changes. Selective HDAC inhibitors will be better able to avoid these types of situations than non-specific inhibitors, and may also be less toxic than pan-HDAC inhibitors. Many potent pan-HDAC inhibitors have already been developed, leaving the development of selective inhibitors at the forefront of HDAC drug development. Certain structural moieties may be added to HDAC inhibitors to give isoform selectivity, and these will be discussed in this review. This review will focus on the applications of selective HDAC inhibitors, inhibitors reported to show selectivity, and the relationship between inhibitor structure and selectivity.

  20. Isoform-targeted regulation of cardiac adenylyl cyclase.

    Science.gov (United States)

    Ishikawa, Yoshihiro

    2003-01-01

    Numerous attempts have been made to develop strategies for regulating the intracellular cyclic AMP signal pharmacologically, with an intention to establish either new medical therapeutic methods or experimental tools. In the past decades, many pharmacological reagents have been identified that regulate this pathway at the level of the receptor. G protein, adenylyl cyclase, cyclic AMP, protein kinase A and phosphodiesterase. Since the cloning of adenylyl cyclase isoforms during the 1990s, investigators including ourselves have tried to find reagents that regulate the activity of this enzyme directly in an isoform-dependent manner. The ultimate goal of developing such reagents would be to regulate the cyclic AMP signal in an organ-dependent manner. Ourselves and other workers have reported that such reagents may vary from a simple cation to kinases. In a more recent study, using the results from crystallographic studies and computer-assisted drug design programs, we have identified subtype-selective regulators of adenylyl cyclase. Such regulators are mostly based upon forskolin, a diterpene compound obtained from Coleus forskolii, that acts directly on adenylyl cyclase to increase the intracellular levels of cyclic AMP. Similarly, novel reagents have been identified that inhibit a specific adenylyl cyclase isoform (e.g. type 5 adenylyl cyclase). Such reagents would potentially provide a new therapeutic strategy to treat hypertension, for example, as well as methods to selectively stimulate or inhibit this adenylyl cyclase isoform, which may be reminiscent of overexpression or knocking out of the cardiac adenylyl cyclase isoform by the use of a pharmacological method.

  1. p53 isoform profiling in glioblastoma and injured brain.

    Science.gov (United States)

    Takahashi, R; Giannini, C; Sarkaria, J N; Schroeder, M; Rogers, J; Mastroeni, D; Scrable, H

    2013-06-27

    The tumor suppressor p53 has been found to be the most commonly mutated gene in human cancers; however, the frequency of p53 mutations varies from 10 to 70% across different cancer types. This variability can partly be explained by inactivating mechanisms aside from direct genomic polymorphisms. The p53 gene encodes 12 isoforms, some of which can modulate full-length p53 activity in cancer. In this study, we characterized p53 isoform expression patterns in glioblastoma, gliosis, non-tumor brain and neural progenitor cells by SDS-PAGE, immunoblot, mass spectrometry and reverse transcription-PCR. We found that the most consistently expressed isoform in glioblastoma, Δ40p53, was uniquely expressed in regenerative processes, such as those involving neural progenitor cells and gliosis compared with tumor samples. Isoform profiling of glioblastoma tissues revealed the presence of both Δ40p53 and full-length p53, neither of which were detected in non-tumor cerebral cortex. Upon xenograft propagation of tumors, p53 levels increased. The variability of overall p53 expression and relative levels of isoforms suggest fluctuations in subpopulations of cells with greater or lesser capacity for proliferation, which can change as the tumor evolves under different growth conditions.

  2. MSD与TPC技术调频遥测方法研究%Research on PCM-FM Technology Based on MSD and TPC

    Institute of Scientific and Technical Information of China (English)

    闫冬; 孙晓锋; 孙大元

    2016-01-01

    调频信号具有很好的抗航天器飞行尾焰干扰能力。在同信噪比下,调频信号解调误码率高于调相信号,所以调频信号解调需要提高增益。多符号检测( MSD)和Turbo乘积( TPC)编码的联合算法可以提高调频信号的增益。介绍了MSD和TPC编码的联合算法和设计结构,根据仿真结果对比了差分解调、 MSD解调、 MSD与TPC编码的联合算法在各种信噪比下的性能指标,证明了该联合算法提高了近7 dB的增益。%The PCM⁃FM signal has better anti⁃interference capability for aerospace⁃craft jetting out the flame.In the same signal⁃noise ratio,the BER of PCM⁃FM signal demodulation is higher than that of phase modulated signal,so it is required to improve the gain of PCM⁃FM signal demodulation.The joint algorithm of multi⁃signal detection (MSD) and Turbo product coding (TPC) can improve the gain of PCM⁃FM signal.This paper introduces the joint algorithm of MSD and TPC coding and design structure.The performance index of differential demodulation,MSD demodulation,as well as MSD and TPC coding joint algoritm are compared in various signal⁃noise ratios, and the results show this joint algorithm can increase the gain of 7 dB.

  3. Neutron Fluence and Energy Reconstruction with the LNE-IRSN/MIMAC Recoil Detector MicroTPC at 27 keV

    Energy Technology Data Exchange (ETDEWEB)

    Maire, D.; Lebreton, L.; Querre, Ph. [Institute for Radioprotection and Nuclear Safety - IRSN, site of Cadarache, 13115 Saint Paul lez Durance (France); Bosson, G.; Guillaudin, O.; Muraz, J.F.; Riffard, Q.; Santos, D. [Laboratoire de Physique Subatomique et de Cosmologie - LPSCCNRSIN2P3/ UJF/INP, 38000 Grenoble (France)

    2015-07-01

    The French Institute for Radiation protection and Nuclear Safety (IRSN), designated by the French Metrology Institute (LNE) for neutron metrology, is developing a time projection chamber using a Micromegas anode: microTPC. This work is carried out in collaboration with the Laboratory of Subatomic Physics and Cosmology (LPSC). The aim is to characterize the energy distribution of neutron fluence in the energy range 8 keV - 5 MeV with a primary procedure. The time projection chambers are gaseous detectors able to measure charged particles energy and to reconstruct their track if a pixelated anode is used. In our case, the gas is used as a (n, p) converter in order to detect neutrons down to few keV. Coming from elastic collisions with neutrons, recoil protons lose a part of their kinetic energy by ionizing the gas. The ionization electrons are drifted toward a pixelated anode (2D projection), read at 50 MHz by a self-triggered electronic system to obtain the third track dimension. The neutron energy is reconstructed event by event thanks to proton scattering angle and proton energy measurements. The scattering angle is deduced from the 3D track. The proton energy is obtained by charge collection measurements, knowing the ionization quenching factor (i.e. the part of proton kinetic energy lost by ionizing the gas). The fluence is calculated thanks to the detected events number and the simulation of the detector response. The μTPC is a new reliable detector able to measure energy distribution of the neutron fluence without unfolding procedure or prior neutron calibration contrary to usual gaseous counters. The microTPC is still being developed and measurements have been carried out at the AMANDE facility, with neutrons energies going from 8 keV to 565 keV. After the context and the μ-TPC working principle presentation, measurements of the neutron energy and fluence at 27 keV and 144 keV are shown and compared to the complete detector response simulation. This work

  4. Neutron fluence and energy reconstruction with the IRSN recoil detector μ-TPC at 27 keV, 144 keV and 565 keV

    Energy Technology Data Exchange (ETDEWEB)

    Maire, D.; Lebreton, L.; Richer, J.P. [IRSN, PRP-HOM, SDE, LMDN, 13115 Saint Paul-Lez-Durance (France); Bosson, G.; Bourrion, O.; Guillaudin, O.; Riffard, Q.; Santos, D. [CNRS/IN2P3-UJF-INPG, LPSC, 38000 Grenoble (France)

    2015-07-01

    The French Institute for Radioprotection and Nuclear Safety (IRSN), associated to the French Metrology Institute (LNE), is developing a time projection chamber using a Micromegas anode: μ-TPC. This work is carried out in collaboration with the Laboratory of Subatomic Physics and Cosmology (LPSC). The aim is to characterize with a primary procedure the energy distribution of neutron fluence in the energy range 8 keV - 1 MeV. The time projection chambers are gaseous detectors, which are able to measure charged particles energy and to reconstruct their track if a pixelated anode is used. In our case, the gas is used as a (n, p) converter in order to detect neutrons down to few keV. Coming from elastic collisions with neutrons, recoil protons lose a part of their kinetic energy by ionizing the gas. The ionization electrons are drifted toward a pixelated anode (2D projection), read at 50 MHz by a self-triggered electronic system to obtain the third track dimension. The neutron energy is reconstructed event by event thanks to proton scattering angle and proton energy measurements. The scattering angle is deduced from the 3D track. The proton energy is obtained by charge collection measurements, knowing the ionization quenching factor (i.e. the part of proton kinetic energy lost by ionizing the gas). The fluence is calculated thanks to the detected events number and the simulated detector response. The μ-TPC is a new reliable detector which enables to measure energy distribution of the neutron fluence without deconvolution or neutron calibration contrary to usual gaseous counters. The μ-TPC is still being developed and measurements have been carried out at the AMANDE facility, with neutrons energies going from 8 keV to 565 keV. After the context and the μ-TPC working principle presentation, measurements of the neutron energy and fluence at 27.2 keV, 144 keV and 565 keV are shown and compared to the complete detector simulation. This work shows the first direct

  5. Laminin isoforms in endothelial and perivascular basement membranes.

    Science.gov (United States)

    Yousif, Lema F; Di Russo, Jacopo; Sorokin, Lydia

    2013-01-01

    Laminins, one of the major functional components of basement membranes, are found underlying endothelium, and encasing pericytes and smooth muscle cells in the vessel wall. Depending on the type of blood vessel (capillary, venule, postcapillary venule, vein or artery) and their maturation state, both the endothelial and mural cell phenotype vary, with associated changes in laminin isoform expression. Laminins containing the α4 and α5 chains are the major isoforms found in the vessel wall, with the added contribution of laminin α2 in larger vessels. We here summarize current data on the precise localization of these laminin isoforms and their receptors in the different layers of the vessel wall, and their potential contribution to vascular homeostasis.

  6. Proteogenomic Analysis Identifies a Novel Human SHANK3 Isoform

    Directory of Open Access Journals (Sweden)

    Fahad Benthani

    2015-05-01

    Full Text Available Mutations of the SHANK3 gene have been associated with autism spectrum disorder. Individuals harboring different SHANK3 mutations display considerable heterogeneity in their cognitive impairment, likely due to the high SHANK3 transcriptional diversity. In this study, we report a novel interaction between the Mutated in colorectal cancer (MCC protein and a newly identified SHANK3 protein isoform in human colon cancer cells and mouse brain tissue. Hence, our proteogenomic analysis identifies a new human long isoform of the key synaptic protein SHANK3 that was not predicted by the human reference genome. Taken together, our findings describe a potential new role for MCC in neurons, a new human SHANK3 long isoform and, importantly, highlight the use of proteomic data towards the re-annotation of GC-rich genomic regions.

  7. Assessment of the efficiency of SimPlate™ total plate count color indicator (TPC CI to quantify mesophilic aerobic microorganisms in pasteurized milk Avaliação da eficiência do SimPlate™ Total Plate Count Color Indicator (TPC CI para enumeração de microrganismos aeróbios mesófilos em leite pasteurizado

    Directory of Open Access Journals (Sweden)

    Luís Augusto Nero

    2002-01-01

    Full Text Available The SimPlate™ TPC CI system is a rapid method to count mesophilic aerobic microorganisms (MAM in foods, based on the use of resazurine to indicate bacterial growth. Its efficiency in pasteurized milk was evaluated using 142 pasteurized milk samples (38 type A, 43 type B and 61 type C collected in Londrina, PR. The standard plating method, using Plate Count Agar (PCA was used for comparison. The plates of both systems were incubated at 35ºC and read after 24h and 48h. The occurrence of false-positive and false-negative wells and the predominant microorganisms in them were also evaluated. The results were compared by simple correlation and mean variance analyses. The correlation (r and mean variance values were 0.6811 and 0.7583 for the results obtained after 24h, respectively, and 0.9126 and 0.0842 for the results obtained after 48h, respectively. These results indicate that the performance of the system increases when the plates are incubated for 48h. When the three types of milk were evaluated separately, these values were 0.9285 and 0.0817 for type A milk, 0.9231 and 0.0466 for type B milk and 0.7209 and 0.1082 for type C milk. These results indicate that the better the quality of the milk the better the performance of SimPlate™ TPC CI. False-negative wells, found more frequently in samples with high MAM counts, were caused by Gram positive microorganisms, poorly detected by the SimPlate™ TPC CI system because they grew slowly and had low reduction capacity. The results indicated a higher efficiency of the SimPlate™ TPC CI system in the reading at 48h.O sistema SimPlate™ TPC CI é um método rápido para enumeração de microrganismos aeróbios mesófilos (MAM em alimentos que utiliza a resazurina como substância indicadora de crescimento bacteriano. Para avaliar sua eficiência em leite pasteurizado, 142 amostras (38 de leite tipo A, 43 de leite tipo B e 61 de leite tipo C foram colhidas em Londrina, PR, e analisadas pelo Sim

  8. Rôles respectifs des isoformes de ferrédoxine-NADP-oxydoréductase dans la cyanobactérie Synechocystis sp. PCC 6803

    OpenAIRE

    Korn, Anja

    2010-01-01

    In photosynthetic organisms, ferredoxin:NADP oxidoreductase (FNR) provides NADPH for CO2 assimilation, but it also utilizes NADPH to provide reduced ferredoxin (Fd). The cyanobacterium Synechocystis sp. strain PCC6803 contains two FNR isoforms, a small (FNRS, 34 kDa) and a large one (FNRL, 46 kDa) that is associated with the phycobilisome (PBS), a light-harvesting complex. We purified a PBS subcomplex comprising FNRL (FNRL-PC) and compared the enzymatic properties of FNRL-PC to FNRS. FNRL-PC ...

  9. TPC data compression

    Energy Technology Data Exchange (ETDEWEB)

    Berger, Jens; Frankenfeld, Ulrich; Lindenstruth, Volker; Plamper, Patrick; Roehrich, Dieter; Schaefer, Erich; W. Schulz, Markus; M. Steinbeck, Timm; Stock, Reinhard; Sulimma, Kolja; Vestboe, Anders; Wiebalck, Arne E-mail: wiebalck@kip.uni-heidelberg.de

    2002-08-21

    In the collisions of ultra-relativistic heavy ions in fixed-target and collider experiments, multiplicities of several ten thousand charged particles are generated. The main devices for tracking and particle identification are large-volume tracking detectors (TPCs) producing raw event sizes in excess of 100 Mbytes per event. With increasing data rates, storage becomes the main limiting factor in such experiments and, therefore, it is essential to represent the data in a way that is as concise as possible. In this paper, we present several compression schemes, such as entropy encoding, modified vector quantization, and data modeling techniques applied on real data from the CERN SPS experiment NA49 and on simulated data from the future CERN LHC experiment ALICE.

  10. TPC data compression

    CERN Document Server

    Berger, Jens; Lindenstruth, Volker; Plamper, Patrick; Röhrich, Dieter; Schafer, Erich; Schulz, M W; Steinbeck, T M; Stock, Reinhard; Sulimma, Kolja; Vestbo, Anders S; Wiebalck, Arne

    2002-01-01

    In the collisions of ultra-relativistic heavy ions in fixed-target and collider experiments, multiplicities of several ten thousand charged particles are generated. The main devices for tracking and particle identification are large-volume tracking detectors (TPCs) producing raw event sizes in excess of 100 Mbytes per event. With increasing data rates, storage becomes the main limiting factor in such experiments and, therefore, it is essential to represent the data in a way that is as concise as possible. In this paper, we present several compression schemes, such as entropy encoding, modified vector quantization, and data modeling techniques applied on real data from the CERN SPS experiment NA49 and on simulated data from the future CERN LHC experiment ALICE.

  11. TPC data compression

    Science.gov (United States)

    Berger, Jens; Frankenfeld, Ulrich; Lindenstruth, Volker; Plamper, Patrick; Röhrich, Dieter; Schäfer, Erich; Schulz, Markus W.; Steinbeck, Timm M.; Stock, Reinhard; Sulimma, Kolja; Vestbø, Anders; Wiebalck, Arne

    2002-08-01

    In the collisions of ultra-relativistic heavy ions in fixed-target and collider experiments, multiplicities of several ten thousand charged particles are generated. The main devices for tracking and particle identification are large-volume tracking detectors (TPCs) producing raw event sizes in excess of 100 Mbytes per event. With increasing data rates, storage becomes the main limiting factor in such experiments and, therefore, it is essential to represent the data in a way that is as concise as possible. In this paper, we present several compression schemes, such as entropy encoding, modified vector quantization, and data modeling techniques applied on real data from the CERN SPS experiment NA49 and on simulated data from the future CERN LHC experiment ALICE.

  12. Autocrine VEGF isoforms differentially regulate endothelial cell behavior

    Directory of Open Access Journals (Sweden)

    Hideki Yamamoto

    2016-09-01

    Full Text Available Vascular endothelial growth factor A (VEGF is involved in all the essential biology of endothelial cells, from proliferation to vessel function, by mediating intercellular interactions and monolayer integrity. It is expressed as three major alternative spliced variants. In mice, these are VEGF120, VEGF164, and VEGF188, each with different affinities for extracellular matrices and cell surfaces, depending on the inclusion of heparin-binding sites, encoded by exons 6 and 7. To determine the role of each VEGF isoform in endothelial homeostasis, we compared phenotypes of primary endothelial cells isolated from lungs of mice expressing single VEGF isoforms in normoxic and hypoxic conditions. The differential expression and distribution of VEGF isoforms affect endothelial cell functions, such as proliferation, adhesion, migration and integrity, which are dependent on the stability of and affinity to VEGF receptor 2 (VEGFR2. We found a correlation between autocrine VEGF164 and VEGFR2 stability, which is also associated with increased expression of proteins involved in cell adhesion. Endothelial cells expressing only VEGF188, which localizes to extracellular matrices or cell surfaces, presented a mesenchymal morphology and weakened monolayer integrity. Cells expressing only VEGF120 lacked stable VEGFR2 and dysfunctional downstream processes, rendering the cells unviable. Endothelial cells expressing these different isoforms in isolation also had differing rates of apoptosis, proliferation, and signaling via nitric oxide (NO synthesis. These data indicate that autocrine signaling of each VEGF isoform has unique functions on endothelial homeostasis and response to hypoxia, due to both distinct VEGF distribution and VEGFR2 stability, which appears to be, at least partly, affected by differential NO production. This study demonstrates that each autocrine VEGF isoform has a distinct effect on downstream functions, namely VEGFR2-regulated endothelial cell

  13. Identification and characterization of novel NuMA isoforms

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Jin, E-mail: petersdu2112@hotmail.com [Key Laboratory for Cell Proliferation and Regulation of the Ministry of Education, Beijing Normal University, Beijing (China); Xu, Zhe [Department of Clinical Laboratory Diagnosis, Beijing Tiantan Hospital, Capital Medical University, Beijing (China); Core Laboratory for Clinical Medical Research, Beijing Tiantan Hospital, Capital Medical University, Beijing (China); He, Dacheng [Key Laboratory for Cell Proliferation and Regulation of the Ministry of Education, Beijing Normal University, Beijing (China); Lu, Guanting, E-mail: guantlv@126.com [Beijing DnaLead Science and Technology Co., LTD, Beijing (China)

    2014-11-21

    Highlights: • Seven NuMA isoforms generated by alternative splicing were categorized into 3 groups: long, middle and short. • Both exons 15 and 16 in long NuMA were “hotspot” for alternative splicing. • Lower expression of short NuMA was observed in cancer cells compared with nonneoplastic controls. • Distinct localization pattern of short isoforms indicated different function from that of long and middle NuMA. - Abstract: The large nuclear mitotic apparatus (NuMA) has been investigated for over 30 years with functions related to the formation and maintenance of mitotic spindle poles during mitosis. However, the existence and functions of NuMA isoforms generated by alternative splicing remains unclear. In the present work, we show that at least seven NuMA isoforms (categorized into long, middle and short groups) generated by alternative splicing from a common NuMA mRNA precursor were discovered in HeLa cells and these isoforms differ mainly at the carboxyl terminus and the coiled-coil domains. Two “hotspot” exons with molecular mass of 3366-nt and 42-nt tend to be spliced during alternative splicing in long and middle groups. Furthermore, full-length coding sequences of long and middle NuMA obtained by using fusion PCR were constructed into GFP-tagged vector to illustrate their cellular localization. Long NuMA mainly localized in the nucleus with absence from nucleoli during interphase and translocated to the spindle poles in mitosis. Middle NuMA displayed the similar cell cycle-dependent distribution pattern as long NuMA. However, expression of NuMA short isoforms revealed a distinct subcellular localization. Short NuMA were present in the cytosol during the whole cycle, without colocalization with mitotic apparatus. These results have allowed us tentatively to explore a new research direction for NuMA’s various functions.

  14. Isolation and structural characterization of different isoforms of the hypusine-containing protein eIF-5A from HeLa cells.

    Science.gov (United States)

    Klier, H; Csonga, R; Joäo, H C; Eckerskorn, C; Auer, M; Lottspeich, F; Eder, J

    1995-11-14

    Posttranslational modification of a specific lysine residue in eukaryotic initiation factor 5A (eIF-5A) is essential for cell viability and proliferation. The product of this modification is hypusine, an amino acid unique to eIF-5A. We have purified and characterized one major and three minor isoforms of human eIF-5A from HeLa cells. The main form, which accounts for approximately 95% of the total eIF-5A, carries hypusine at position 50 and is amino-terminally acetylated as determined by amino acid composition analysis and electrospray ionization mass spectrometry. Analytical gel filtration indicates that this protein variant possesses a native apparent molecular weight that lies between that expected for a monomeric and dimeric form. Nevertheless, several experiments confirm this protein to be monomeric. It is further shown that eIF-5A have well-defined secondary structure. Both the far-UV circular dichroism spectrum as well as secondary structure predictions using different algorithms suggest this protein to have predominantly beta-sheet structure. Two plausible models for the packing of the secondary structure elements are presented. In contrast to the main form, all three minor isoforms of eIF-5A are characterized by acetylation of the epsilon-amino group of lysine at position 47. The minor isoforms are distinguishable by their state of modification of the lysine residue at position 50. Whereas the main form occurs in both the cytoplasmic and the nuclear fraction of HeLa cells, the minor isoforms were not detectable in the preparation of the nuclear fraction. Therefore, acetylation of lysine at position 47 might play a controlling role in the distribution of the minor isoforms to the nucleus.

  15. Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.

    Directory of Open Access Journals (Sweden)

    Jinte Middeldorp

    Full Text Available The human GFAP splice variants GFAPDelta164 and GFAPDeltaexon6 both result in a GFAP protein isoform with a unique out-of-frame carboxy-terminus that can be detected by the GFAP+1 antibody. We previously reported that GFAP+1 was expressed in astrocytes and in degenerating neurons in Alzheimer's disease brains. In this study we aimed at further investigating the neuronal GFAP+1 expression and we started by affinity purifying the GFAP+1 antibody. The purified antibody resulted in a loss of neuronal GFAP+1 signal, although other antibodies directed against the amino- and carboxy-terminus of GFAPalpha still revealed GFAP-immunopositive neurons, as described before. With an in-depth analysis of a western blot, followed by mass spectrometry we discovered that the previously detected neuronal GFAP+1 expression was due to cross-reactivity of the antibody with neurofilament-L (NF-L. This was confirmed by double-label fluorescent immunohistochemistry and western blotting with the unpurified GFAP+1 antibody and an antibody against NF-L. Our data imply that NF-L can accumulate in some tangle-like structures in Alzheimer brains. More importantly, the purified GFAP+1 antibody clearly revealed a specific subtype of astrocytes in the adult human brain. These large astrocytes are present throughout the brain, e.g., along the subventricular zone, in the hippocampus, in the striatum and in the spinal cord of controls, Alzheimer, and Parkinson patients. The presence of a specific GFAP-isoform suggests a specialized function of these astrocytes.

  16. Specific Human Astrocyte Subtype Revealed by Affinity Purified GFAP+1 Antibody; Unpurified Serum Cross-Reacts with Neurofilament-L in Alzheimer

    Science.gov (United States)

    Middeldorp, Jinte; van den Berge, Simone A.; Aronica, Eleonora; Speijer, Dave; Hol, Elly M.

    2009-01-01

    The human GFAP splice variants GFAPΔ164 and GFAPΔexon6 both result in a GFAP protein isoform with a unique out-of-frame carboxy-terminus that can be detected by the GFAP+1 antibody. We previously reported that GFAP+1 was expressed in astrocytes and in degenerating neurons in Alzheimer's disease brains. In this study we aimed at further investigating the neuronal GFAP+1 expression and we started by affinity purifying the GFAP+1 antibody. The purified antibody resulted in a loss of neuronal GFAP+1 signal, although other antibodies directed against the amino- and carboxy-terminus of GFAPα still revealed GFAP-immunopositive neurons, as described before. With an in-depth analysis of a western blot, followed by mass spectrometry we discovered that the previously detected neuronal GFAP+1 expression was due to cross-reactivity of the antibody with neurofilament-L (NF-L). This was confirmed by double-label fluorescent immunohistochemistry and western blotting with the unpurified GFAP+1 antibody and an antibody against NF-L. Our data imply that NF-L can accumulate in some tangle-like structures in Alzheimer brains. More importantly, the purified GFAP+1 antibody clearly revealed a specific subtype of astrocytes in the adult human brain. These large astrocytes are present throughout the brain, e.g., along the subventricular zone, in the hippocampus, in the striatum and in the spinal cord of controls, Alzheimer, and Parkinson patients. The presence of a specific GFAP-isoform suggests a specialized function of these astrocytes. PMID:19888461

  17. Properties of purified recombinant human polyamine oxidase, PAOh1/SMO.

    Science.gov (United States)

    Wang, Yanlin; Murray-Stewart, Tracy; Devereux, Wendy; Hacker, Amy; Frydman, Benjamin; Woster, Patrick M; Casero, Robert A

    2003-05-16

    The discovery of an inducible oxidase whose apparent substrate preference is spermine indicates that polyamine catabolism is more complex than that originally proposed. To facilitate the study of this enzyme, the purification and characterization of the recombinant human PAOh1/SMO polyamine oxidase are reported. Purified PAOh1/SMO oxidizes both spermine (K(m)=1.6 microM) and N(1)-acetylspermine (K(m)=51 microM), but does not oxidize spermidine. The purified human enzyme also does not oxidize eight representative antitumor polyamine analogues; however, specific oligamine analogues were found to be potent inhibitors of the oxidation of spermine by PAOh1/SMO. The results of these studies are consistent with the hypothesis that PAOh1/SMO represents a new addition to the polyamine metabolic pathway that may represent a new target for antineoplastic drug development.

  18. Thermal inactivation kinetics of partially purified mango pectin methylesterase

    Directory of Open Access Journals (Sweden)

    Claudio Alonso DÍAZ-CRUZ

    2016-01-01

    Full Text Available Abstract Kinetic parameters of thermal inactivation of pectin methylesterase (PME in a partially purified mango enzyme extract were determined. The PME of mango partially purified by salting out showed different patterns of thermal inactivation, indicating the presence of a thermostable fraction at 70 °C and a thermolabile fraction at lower temperatures. The inactivation of the thermostable fraction exhibited a linear behavior that yielded a z-value of 9.44 °C and an activation energy (Ea of 245.6 kJ mol-1 K-1 using the Arrhenius model. The thermostable mango PME fraction represented 17% of total crude enzyme extract, which emphasizes the importance of residual enzyme activity after heat treatment.

  19. 42 CFR 84.171 - Non-powered air-purifying particulate respirators; required components.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Non-powered air-purifying particulate respirators... PROTECTIVE DEVICES Non-Powered Air-Purifying Particulate Respirators § 84.171 Non-powered air-purifying particulate respirators; required components. (a) Each non-powered air-purifying particulate...

  20. 42 CFR 84.170 - Non-powered air-purifying particulate respirators; description.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Non-powered air-purifying particulate respirators... DEVICES Non-Powered Air-Purifying Particulate Respirators § 84.170 Non-powered air-purifying particulate respirators; description. (a) Non-powered air-purifying particulate respirators utilize the wearer's...

  1. Isolation of Microorganisms Able To Metabolize Purified Natural Rubber

    OpenAIRE

    Heisey, R. M.; Papadatos, S

    1995-01-01

    Bacteria able to grow on purified natural rubber in the absence of other organic carbon sources were isolated from soil. Ten isolates reduced the weight of vulcanized rubber from latex gloves by >10% in 6 weeks. Scanning electron microscopy clearly revealed the ability of the microorganisms to colonize, penetrate, and dramatically alter the physical structure of the rubber. The rubber-metabolizing bacteria were identified on the basis of fatty acid profiles and cell wall characteristics. Seve...

  2. Survey of Air Purifier Market Acceptance in China

    OpenAIRE

    Yang, Shan

    2016-01-01

    In recent years, air cleaner products have drawn a wide attention due to the extensive concern of air pollution in China. The study aims at research market acceptance of air purifiers. Meanwhile, an outlook of present market and competitive environment were introduced for driving forces of the research as background knowledge. In this thesis, a theoretical framework was designed to express the theory of customer acceptance, which provided theoretical support for the analysis process in th...

  3. Proteomic analysis of purified Newcastle disease virus particles

    Directory of Open Access Journals (Sweden)

    Ren Xiangpeng

    2012-05-01

    Full Text Available Abstract Background Newcastle disease virus (NDV is an enveloped RNA virus, bearing severe economic losses to the poultry industry worldwide. Previous virion proteomic studies have shown that enveloped viruses carry multiple host cellular proteins both internally and externally during their life cycle. To address whether it also occurred during NDV infection, we performed a comprehensive proteomic analysis of highly purified NDV La Sota strain particles. Results In addition to five viral structural proteins, we detected thirty cellular proteins associated with purified NDV La Sota particles. The identified cellular proteins comprised several functional categories, including cytoskeleton proteins, annexins, molecular chaperones, chromatin modifying proteins, enzymes-binding proteins, calcium-binding proteins and signal transduction-associated proteins. Among these, three host proteins have not been previously reported in virions of other virus families, including two signal transduction-associated proteins (syntenin and Ras small GTPase and one tumor-associated protein (tumor protein D52. The presence of five selected cellular proteins (i.e., β-actin, tubulin, annexin A2, heat shock protein Hsp90 and ezrin associated with the purified NDV particles was validated by Western blot or immunogold labeling assays. Conclusions The current study presented the first standard proteomic profile of NDV. The results demonstrated the incorporation of cellular proteins in NDV particles, which provides valuable information for elucidating viral infection and pathogenesis.

  4. Proteomic analysis of purified coronavirus infectious bronchitis virus particles

    Directory of Open Access Journals (Sweden)

    Shu Dingming

    2010-06-01

    Full Text Available Abstract Background Infectious bronchitis virus (IBV is the coronavirus of domestic chickens causing major economic losses to the poultry industry. Because of the complexity of the IBV life cycle and the small number of viral structural proteins, important virus-host relationships likely remain to be discovered. Toward this goal, we performed two-dimensional gel electrophoresis fractionation coupled to mass spectrometry identification approaches to perform a comprehensive proteomic analysis of purified IBV particles. Results Apart from the virus-encoded structural proteins, we detected 60 host proteins in the purified virions which can be grouped into several functional categories including intracellular trafficking proteins (20%, molecular chaperone (18%, macromolcular biosynthesis proteins (17%, cytoskeletal proteins (15%, signal transport proteins (15%, protein degradation (8%, chromosome associated proteins (2%, ribosomal proteins (2%, and other function proteins (3%. Interestingly, 21 of the total host proteins have not been reported to be present in virions of other virus families, such as major vault protein, TENP protein, ovalbumin, and scavenger receptor protein. Following identification of the host proteins by proteomic methods, the presence of 4 proteins in the purified IBV preparation was verified by western blotting and immunogold labeling detection. Conclusions The results present the first standard proteomic profile of IBV and may facilitate the understanding of the pathogenic mechanisms.

  5. Crystallization and preliminary X-ray crystallographic analysis of latent isoform PPO4 mushroom (Agaricus bisporus) tyrosinase.

    Science.gov (United States)

    Mauracher, Stephan Gerhard; Molitor, Christian; Al-Oweini, Rami; Kortz, Ulrich; Rompel, Annette

    2014-02-01

    Tyrosinase exhibits catalytic activity for the ortho-hydroxylation of monophenols to diphenols as well as their subsequent oxidation to quinones. Owing to polymerization of these quinones, brown-coloured high-molecular-weight compounds called melanins are generated. The latent precursor form of polyphenol oxidase 4, one of the six tyrosinase isoforms from Agaricus bisporus, was purified to homogeneity and crystallized. The obtained crystals belonged to space group C121 (two molecules per asymmetric unit) and diffracted to 2.78 Å resolution. The protein only formed crystals under low-salt conditions using the 6-tungstotellurate(VI) salt Na6[TeW6O24] · 22H2O as a co-crystallization agent.

  6. A Prototype Combination TPC Cherenkov Detector with GEM Readout for Tracking and Particle Identification and its Potential Use at an Electron Ion Collider

    CERN Document Server

    Woody, Craig; Majka, Richard; Phipps, Michael; Purschke, Martin; Smirnov, Nikolai

    2015-01-01

    A prototype detector is being developed which combines the functions of a Time Projection Chamber for charged particle tracking and a Cherenkov detector for particle identification. The TPC consists of a 10x10x10 cm3 drift volume where the charge is drifted to a 10x10 cm2 triple GEM detector. The charge is measured on a readout plane consisting of 2x10 mm2 chevron pads which provide a spatial resolution ~ 100 microns per point in the chevron direction along with dE/dx information. The Cherenkov portion of the detector consists of a second 10x10 cm2 triple GEM with a photosensitive CsI photocathode on the top layer. This detector measures Cherenkov light produced in the drift gas of the TPC by high velocity particles which are above threshold. CF4 or CF4 mixtures will be used as the drift gas which are highly transparent to UV light and can provide excellent efficiency for detecting Cherenkov photons. The drift gas is also used as the operating gas for both GEM detectors. The prototype detector has been constr...

  7. Accurate gamma and MeV-electron track reconstruction with an ultra-low diffusion Xenon/TMA TPC at 10 atmospheres

    CERN Document Server

    González-Díaz, Diego; Borges, F.I.G.; Camargo, M.; Cárcel, S.; Cebrián, S.; Cervera, A.; Conde, C.A.N.; Dafni, T.; Díaz, J.; Esteve, R.; Fernandes, L.M.P.; Ferrario, P.; Ferreira, A.L.; Freitas, E.D.C.; Gehman, V.M.; Goldschmidt, A.; Gómez-Cadenas, J.J.; Gutiérrez, R.M.; Hauptman, J.; Hernando Morata, J.A.; Herrera, D.C.; Irastorza, I.G.; Labarga, L.; Laing, A.; Liubarsky, I.; Lopez-March, N.; Lorca, D.; Losada, M.; Luzón, G.; Marí, A.; Martín-Albo, J.; Martínez-Lema, G.; Martínez, A.; Miller, T.; Monrabal, F.; Monserrate, M.; Monteiro, C.M.B.; Mora, F.J.; Moutinho, L.M.; Muñoz Vidal, J.; Nebot-Guinot, M.; Nygren, D.; Oliveira, C.A.B.; Pérez, J.; Pérez Aparicio, J.L.; Querol, M.; Renner, J.; Ripoll, L.; Rodríguez, J.; Santos, F.P.; dos Santos, J.M.F.; Serra, L.; Shuman, D.; Simón, A.; Sofka, C.; Sorel, M.; Toledo, J.F.; Torrent, J.; Tsamalaidze, Z.; Veloso, J.F.C.A.; Villar, J.A.; Webb, R.; White, J.T.; Yahlali, N.; Azevedo, C.; Aznar, F.; Calvet, D.; Castel, J.; Ferrer-Ribas, E.; García, J.A.; Giomataris, I.; Gómez, H.; Iguaz, F.J.; Lagraba, A.; Le Coguie, A.; Mols, J.P.; Şahin, Ö.; Rodríguez, A.; Ruiz-Choliz, E.; Segui, L.; Tomás, A.; Veenhof, R.

    2015-01-01

    We report the performance of a 10 atm Xenon/trimethylamine time projection chamber (TPC) for the detection of X-rays (30 keV) and gamma-rays (0.511-1.275 MeV) in conjunction with the accurate tracking of the associated electrons. When operated at such a high pressure and in 1%-admixtures, trimethylamine (TMA) endows Xenon with an extremely low electron diffusion (1.3 +-0.13 mm-sigma (longitudinal), 0.8 +-0.15 mm-sigma (transverse) along 1 m drift) besides forming a convenient Penning-Fluorescent mixture. The TPC, that houses 1.1 kg of gas in its active volume, operated continuously for 100 live-days in charge amplification mode. The readout was performed through the recently introduced microbulk Micromegas technology and the AFTER chip, providing a 3D voxelization of 8mm x 8mm x 1.2mm for approximately 10 cm/MeV-long electron tracks. This work was developed as part of the R&D program of the NEXT collaboration for future detector upgrades in the search of the 0bbnu decay in 136Xe, specifically those based ...

  8. Firmware Development and Integration for ALICE TPC and PHOS Front-end Electronics A Trigger Based Readout and Control System operating in a Radiation Environment

    CERN Document Server

    AUTHOR|(CDS)2068589; Rohrich, Dieter

    2008-01-01

    The readout electronics in PHOS and TPC - two of the major detectors of the ALICE experiment at the LHC - consist of a set of Front End Cards (FECs) that digitize, process and buffer the data from the detector sensors. The FECs are connected to a Readout Control Unit (RCU) via two sets of custom made PCB backplanes. For PHOS, 28 FECs are connected to one RCU, while for TPC the number is varying from 18 to 25 FECs depending on location. The RCU is in charge of the data readout, including reception and distribution of triggers and in moving the data from the FECs to the Data Acquisition System. In addition it does low level control tasks. The RCU consists of an RCU Motherboard that hosts a Detector Control System (DCS) board and a Source Interface Unit. The DCS board is an embedded computer running Linux that controls the readout electronics. All the mentioned devices are implemented in commercial grade SRAM based Field Programmable Gate Arrays (FPGAs). Even if these devices are not very radiation tolerant, the...

  9. First Demonstration of Imaging Cosmic Muons in a Two-Phase Liquid Argon TPC using an EMCCD Camera and a THGEM

    CERN Document Server

    Mavrokoridis, K; McCormick, K J; Paudyal, P; Roberts, A; Smith, N A; Touramanis, C

    2015-01-01

    Colossal two-phase Liquid Argon Time Projection Chambers (LAr TPCs) are a proposed option for future long-baseline neutrino experiments. This study illustrates the feasibility of using an EMCCD camera to capture light induced by single cosmic events in a two-phase LAr TPC employing a THGEM. An Andor iXon Ultra 897 EMCCD camera was externally mounted via a borosilicate glass viewport on the Liverpool two-phase LAr TPC. The camera successfully captured the secondary scintillation light produced at the THGEM holes that had been induced by cosmic events. The light collection capability of the camera for various EMCCD gains was assessed. For a THGEM gain of 64 and an EMCCD gain of 1000, clear images were captured with an average signal-to-noise ratio of 6. Preliminary 3D reconstruction of straight cosmic muon tracks has been performed by combining the camera images, PMT signals and THGEM charge data. Reconstructed cosmic muon tracks were used to determine THGEM gain and to calibrate the intensity levels of the EMC...

  10. Design of the cryogenic systems for the Near and Far LAr-TPC detectors of the Short-Baseline Neutrino program (SBN) at Fermilab

    Energy Technology Data Exchange (ETDEWEB)

    Geynisman, M. [Fermilab; Bremer, J. [CERN; Chalifour, M. [CERN; Delaney, M. [Fermilab; Dinnon, M. [Fermilab; Doubnik, R. [Fermilab; Hentschel, S. [Fermilab; Kim, M. J. [Fermilab; Montanari, C. [INFN, Pavia; Monatanari, D. [Fermilab; Nichols, T. [Fermilab; Norris, B. [Fermilab; Sarychev, M. [Fermilab; Schwartz, F. [Fermilab; Tillman, J. [Fermilab; Zuckerbrot, M. [Fermilab

    2017-08-31

    The Short-Baseline Neutrino (SBN) physics program at Fermilab and Neutrino Platform (NP) at CERN are part of the international Neutrino Program leading to the development of Long-Baseline Neutrino Facility/Deep Underground Neutrino Experiment (LBNF/DUNE) science project. The SBN program consisting of three Liquid Argon Time Projection Chamber (LAr-TPC) detectors positioned along the Booster Neutrino Beam (BNB) at Fermilab includes an existing detector known as MicroBooNE (170-ton LAr-TPC) plus two new experiments known as SBN’s Near Detector (SBND, ~260 tons) and SBN’s Far Detector (SBN-FD, ~760 tons). All three detectors have distinctly different design of their cryostats thus defining specific requirements for the cryogenic systems. Fermilab has already built two new facilities to house SBND and SBN-FD detectors. The cryogenic systems for these detectors are in various stages of design and construction with CERN and Fermilab being responsible for delivery of specific sub-systems. This contribution presents specific design requirements and typical implementation solutions for each sub-system of the SBND and SBN-FD cryogenic systems.

  11. Systematically differentiating functions for alternatively spliced isoforms through integrating RNA-seq data.

    Science.gov (United States)

    Eksi, Ridvan; Li, Hong-Dong; Menon, Rajasree; Wen, Yuchen; Omenn, Gilbert S; Kretzler, Matthias; Guan, Yuanfang

    2013-01-01

    Integrating large-scale functional genomic data has significantly accelerated our understanding of gene functions. However, no algorithm has been developed to differentiate functions for isoforms of the same gene using high-throughput genomic data. This is because standard supervised learning requires 'ground-truth' functional annotations, which are lacking at the isoform level. To address this challenge, we developed a generic framework that interrogates public RNA-seq data at the transcript level to differentiate functions for alternatively spliced isoforms. For a specific function, our algorithm identifies the 'responsible' isoform(s) of a gene and generates classifying models at the isoform level instead of at the gene level. Through cross-validation, we demonstrated that our algorithm is effective in assigning functions to genes, especially the ones with multiple isoforms, and robust to gene expression levels and removal of homologous gene pairs. We identified genes in the mouse whose isoforms are predicted to have disparate functionalities and experimentally validated the 'responsible' isoforms using data from mammary tissue. With protein structure modeling and experimental evidence, we further validated the predicted isoform functional differences for the genes Cdkn2a and Anxa6. Our generic framework is the first to predict and differentiate functions for alternatively spliced isoforms, instead of genes, using genomic data. It is extendable to any base machine learner and other species with alternatively spliced isoforms, and shifts the current gene-centered function prediction to isoform-level predictions.

  12. Systematically differentiating functions for alternatively spliced isoforms through integrating RNA-seq data.

    Directory of Open Access Journals (Sweden)

    Ridvan Eksi

    Full Text Available Integrating large-scale functional genomic data has significantly accelerated our understanding of gene functions. However, no algorithm has been developed to differentiate functions for isoforms of the same gene using high-throughput genomic data. This is because standard supervised learning requires 'ground-truth' functional annotations, which are lacking at the isoform level. To address this challenge, we developed a generic framework that interrogates public RNA-seq data at the transcript level to differentiate functions for alternatively spliced isoforms. For a specific function, our algorithm identifies the 'responsible' isoform(s of a gene and generates classifying models at the isoform level instead of at the gene level. Through cross-validation, we demonstrated that our algorithm is effective in assigning functions to genes, especially the ones with multiple isoforms, and robust to gene expression levels and removal of homologous gene pairs. We identified genes in the mouse whose isoforms are predicted to have disparate functionalities and experimentally validated the 'responsible' isoforms using data from mammary tissue. With protein structure modeling and experimental evidence, we further validated the predicted isoform functional differences for the genes Cdkn2a and Anxa6. Our generic framework is the first to predict and differentiate functions for alternatively spliced isoforms, instead of genes, using genomic data. It is extendable to any base machine learner and other species with alternatively spliced isoforms, and shifts the current gene-centered function prediction to isoform-level predictions.

  13. ApoE isoform-dependent changes in hippocampal synaptic function

    Directory of Open Access Journals (Sweden)

    Sullivan Patrick M

    2009-05-01

    Full Text Available Abstract The lipoprotein receptor system in the hippocampus is intimately involved in the modulation of synaptic transmission and plasticity. The association of specific apoE isoform expression with human neurodegenerative disorders has focused attention on the role of these apoE isoforms in lipoprotein receptor-dependent synaptic modulation. In the present study, we used the apoE2, apoE3 and apoE4 targeted replacement (TR mice along with recombinant human apoE isoforms to determine the role of apoE isoforms in hippocampus area CA1 synaptic function. While synaptic transmission is unaffected by apoE isoform, long-term potentiation (LTP is significantly enhanced in apoE4 TR mice versus apoE2 TR mice. ApoE isoform-dependent differences in LTP induction require NMDA-receptor function, and apoE isoform expression alters activation of both ERK and JNK signal transduction. Acute application of specific apoE isoforms also alters LTP induction while decreasing NMDA-receptor mediated field potentials. Furthermore, acute apoE isoform application does not have the same effects on ERK and JNK activation. These findings demonstrate specific, isoform-dependent effects of human apoE isoforms on adult hippocampus synaptic plasticity and highlight mechanistic differences between chronic apoE isoform expression and acute apoE isoform exposure.

  14. Direct binding of specific AUF1 isoforms to tandem zinc finger domains of tristetraprolin (TTP) family proteins.

    Science.gov (United States)

    Kedar, Vishram P; Zucconi, Beth E; Wilson, Gerald M; Blackshear, Perry J

    2012-02-17

    Tristetraprolin (TTP) is the prototype of a family of CCCH tandem zinc finger proteins that can bind to AU-rich elements in mRNAs and promote their decay. TTP binds to mRNA through its central tandem zinc finger domain; it then promotes mRNA deadenylation, considered to be the rate-limiting step in eukaryotic mRNA decay. We found that TTP and its related family members could bind to certain isoforms of another AU-rich element-binding protein, HNRNPD/AUF1, as well as a related protein, laAUF1. The interaction domain within AUF1p45 appeared to be a C-terminal "GY" region, and the interaction domain within TTP was the tandem zinc finger domain. Surprisingly, binding of AUF1p45 to TTP occurred even with TTP mutants that lacked RNA binding activity. In cell extracts, binding of AUF1p45 to TTP potentiated TTP binding to ARE-containing RNA probes, as determined by RNA gel shift assays; AUF1p45 did not bind to the RNA probes under these conditions. Using purified, recombinant proteins and a synthetic RNA target in FRET assays, we demonstrated that AUF1p45, but not AUF1p37, increased TTP binding affinity for RNA ∼5-fold. These data suggest that certain isoforms of AUF1 can serve as "co-activators" of TTP family protein binding to RNA. The results raise interesting questions about the ability of AUF1 isoforms to regulate the mRNA binding and decay-promoting activities of TTP and its family members as well as the ability of AUF1 proteins to serve as possible physical links between TTP and other mRNA decay proteins and structures.

  15. Isolation and characterization of multiple abundant lipid transfer protein isoforms in developing sesame (Sesamum indicum L.) seeds.

    Science.gov (United States)

    Choi, Ah Mi; Lee, Saet Buyl; Cho, Sung Ho; Hwang, Inhwan; Hur, Cheol-Goo; Suh, Mi Chung

    2008-02-01

    Sesame (Sesamum indicum) is an important oilseed crop; approximately 50% of the seed dry weight is storage oil. In a previous report, developing sesame seed expressed sequence tags (ESTs) revealed that ESTs encoding lipid transfer protein (LTPs) were one of the most abundant groups of sesame ESTs. LTP functions in the transfer of wax or cutin monomers and in the defense response against pathogen attack. To study the biological role of the abundant LTP isoforms in developing seeds, 122 ESTs out of 3328 sesame ESTs were analyzed against Arabidopsis and rice proteome databases. LTP fraction, which was partially purified from developing sesame seeds, actively transferred fluorescent phospholipids and bound to fatty acids. Full-length cDNAs of five out of 21 LTP isoforms were isolated and named SiLTP1-SiLTP5. The predicted amino acid sequences of the five SiLTPs harbor typical characteristics of LTPs, including conserved arrangement of cysteine residues. Northern blot analysis revealed that the five SiLTP isoforms were most abundantly expressed in developing seeds, but were also detected in flower tissues. Also, SiLTP3 and SiLTP4 transcripts were expressed in leaves and seed-pot walls, respectively. In addition, SiLTP2 and SiLTP4 transcripts were significantly induced in 6-day-old sesame seedlings by application of NaCl, mannitol, and abscisic acid (ABA). Transient expression of green fluorescent protein (GFP)-fusion constructs in Arabidopsis protoplasts revealed that SiLTP1 and SiLTP2 were secreted by different pathways. Taken together, the abundant LTPs in developing sesame seeds are involved in lipid transfer into the extracellular matrix. Possible biological roles of SiLTPs related to organ-specific expression and abiotic stresses are discussed.

  16. Herman Feshbach Prize in Theoretical Nuclear Physics Xiangdong Ji, University of Maryland PandaX-III: high-pressure gas TPC for Xe136 neutrinoless double beta decay at CJPL

    Science.gov (United States)

    Ji, Xiangdong; PandaX-III Collaboration

    2016-03-01

    The PandaX-III in China's Jinping Underground Lab is a new neutrinoless double beta decay experiment using Xe136 high-pressure gas TPC. The first phase of the experiment uses a 4 m3 gas detector with symmetric Micromegas charge readout planes. The gas TPC allows full reconstruction of the event topology, capable of distinguishing the two electron events from gamma background with high confidence level. The energy resolution can reach about 3% FWHM at the beta decay Q-value. The detector construction and the experimental lab is currently under active development. In this talk, the current status and future plan are reported.

  17. Conformational Flexibility Differentiates Naturally Occurring Bet v 1 Isoforms.

    Science.gov (United States)

    Grutsch, Sarina; Fuchs, Julian E; Ahammer, Linda; Kamenik, Anna S; Liedl, Klaus R; Tollinger, Martin

    2017-06-03

    The protein Bet v 1 represents the main cause for allergic reactions to birch pollen in Europe and North America. Structurally homologous isoforms of Bet v 1 can have different properties regarding allergic sensitization and Th2 polarization, most likely due to differential susceptibility to proteolytic cleavage. Using NMR relaxation experiments and molecular dynamics simulations, we demonstrate that the initial proteolytic cleavage sites in two naturally occurring Bet v 1 isoforms, Bet v 1.0101 (Bet v 1a) and Bet v 1.0102 (Bet v 1d), are conformationally flexible. Inaccessible cleavage sites in helices and strands are highly flexible on the microsecond-millisecond time scale, whereas those located in loops display faster nanosecond-microsecond flexibility. The data consistently show that Bet v 1.0102 is more flexible and conformationally heterogeneous than Bet v 1.0101. Moreover, NMR hydrogen-deuterium exchange measurements reveal that the backbone amides in Bet v 1.0102 are significantly more solvent exposed, in agreement with this isoform's higher susceptibility to proteolytic cleavage. The differential conformational flexibility of Bet v 1 isoforms, along with the transient exposure of inaccessible sites to the protein surface, may be linked to proteolytic susceptibility, representing a potential structure-based rationale for the observed differences in Th2 polarization and allergic sensitization.

  18. Protein tyrosine phosphatase PTPRR isoforms in cellular signaling and trafficking

    NARCIS (Netherlands)

    Dilaver, Gönül

    2005-01-01

    Previous work has revealed the existence of two Protein Tyrosine Phosphatases in mouse, PTPBR7 and PTP-SL, that were in part identical, suggesting that they originated from the same gene, termed Ptprr (1,5,6). In this thesis, I report on the characterization of the various PTPRR isoforms in neuronal

  19. Murine Sirt3 protein isoforms have variable half-lives

    Science.gov (United States)

    Sirt3 is a NAD+-dependent protein deacetylase mainly localized in mitochondria. Recent studies indicate that the murine Sirt3 gene expresses different transcript variants resulting in three possible Sirt3 protein isoforms with variable lengths at the N-terminus: M1 (aa 1-334), M2 (aa 15-334), and M3...

  20. Isoforms of transferrin in psoriasis patients abusing alcohol

    NARCIS (Netherlands)

    P. Hoefkens (Peter); E.M. Higgins; R.J. Ward (Roberta); H.G. van Eijk (Henk)

    1997-01-01

    textabstractThe different isoforms of transferrin have been quantified by isoelectric focusing in the sera of psoriasis patients with and without a history of abusing alcohol. In both male and female psoriasis subjects abusing alcohol, there were significant increases in the 2-sial

  1. Differential water permeability and regulation of three aquaporin 4 isoforms

    DEFF Research Database (Denmark)

    Fenton, Robert A.; Moeller, Hanne B; Zelenina, Marina

    2010-01-01

    Aquaporin 4 (AQP4) is expressed in the perivascular glial endfeet and is an important pathway for water during formation and resolution of brain edema. In this study, we examined the functional properties and relative unit water permeability of three functional isoforms of AQP4 expressed in the b...

  2. Bacteria-Induced Dscam Isoforms of the Crustacean, Pacifastacus leniusculus.

    Directory of Open Access Journals (Sweden)

    Apiruck Watthanasurorot

    2011-06-01

    Full Text Available The Down syndrome cell adhesion molecule, also known as Dscam, is a member of the immunoglobulin super family. Dscam plays an essential function in neuronal wiring and appears to be involved in innate immune reactions in insects. The deduced amino acid sequence of Dscam in the crustacean Pacifastacus leniusculus (PlDscam, encodes 9(Ig-4(FNIII-(Ig-2(FNIII-TM and it has variable regions in the N-terminal half of Ig2 and Ig3 and the complete Ig7 and in the transmembrane domain. The cytoplasmic tail can generate multiple isoforms. PlDscam can generate more than 22,000 different unique isoforms. Bacteria and LPS injection enhanced the expression of PlDscam, but no response in expression occurred after a white spot syndrome virus (WSSV infection or injection with peptidoglycans. Furthermore, PlDscam silencing did not have any effect on the replication of the WSSV. Bacterial specific isoforms of PlDscam were shown to have a specific binding property to each tested bacteria, E. coli or S. aureus. The bacteria specific isoforms of PlDscam were shown to be associated with bacterial clearance and phagocytosis in crayfish.

  3. Distinct Functions of Endophilin Isoforms in Synaptic Vesicle Endocytosis

    Directory of Open Access Journals (Sweden)

    Jifeng Zhang

    2015-01-01

    Full Text Available Endophilin isoforms perform distinct characteristics in their interactions with N-type Ca2+ channels and dynamin. However, precise functional differences for the endophilin isoforms on synaptic vesicle (SV endocytosis remain unknown. By coupling RNA interference and electrophysiological recording techniques in cultured rat hippocampal neurons, we investigated the functional differences of three isoforms of endophilin in SV endocytosis. The results showed that the amplitude of normalized evoked excitatory postsynaptic currents in endophilin1 knockdown neurons decreased significantly for both single train and multiple train stimulations. Similar results were found using endophilin2 knockdown neurons, whereas endophilin3 siRNA exhibited no change compared with control neurons. Endophilin1 and endophilin2 affected SV endocytosis, but the effect of endophilin1 and endophilin2 double knockdown was not different from that of either knockdown alone. This result suggested that endophilin1 and endophilin2 functioned together but not independently during SV endocytosis. Taken together, our results indicate that SV endocytosis is sustained by endophilin1 and endophilin2 isoforms, but not by endophilin3, in primary cultured hippocampal neurons.

  4. Identification and characterization of a novel isoform of hepatopoietin

    Institute of Scientific and Technical Information of China (English)

    Jun Lu; Wang-Xiang Xu; Yi-Qun Zhan; Xiao-Lin Cui; Wei-Min Cai; Fu-Chu He; Xiao-Ming Yang

    2002-01-01

    AIM: To isolate a novel isoform of human HPO (HPO-205)human fetal liver Marathon-reedy cDNA andcharacterize its primary biological function.METHODS: 5'-RACE (rapid amplification of cDNA 5' ends)was used to isolate a novel isoform of hHPO in this paperThe constructed pcDNAHPO-205, pcDNAHPO and pcDNA eukaryotic expression vectors were respectively transfectedby lipofectamine method and the stimulation of DNAsynthesis was observed by 3H-TdR incorporation assay.Proteins extracted from different cells were analyzed byWestern blot.RESULTS: A novel isoform of hHPO (HPO-205) encoding a205 amino acid ORF corresponding to a translatedproduction of 23 kDa was isolated and distinguished fromthe previous HPO that lacked the N-terminal 80 amino acids.The dnse-dspendent stimulation of DNA synthesis of HepG2hepatoma cells by HPO-205 demonstrated its similarbiological activity with HPO in vitro. The level of MAPK(Mitogen-activated protein kinase) phnsphorylarion byWestern blot analysis revealed that HPO-205 might have thestronger activity of stimulating hepatic cell proliferation thanthat of HPO.CONCLUSION: A novel isoform of hHPO (HPO-205) wasisolated from hepatic-derived cells. The comparison of HPO-205 and HPO will lead to a new insight into the structure andfunction of hHPO, and provide the new way of thinking todeeply elucidate the biological roles of HPO/ALR.

  5. Efficient production of active recombinant Candida rugosa LIP3 lipase in Pichia pastoris and biochemical characterization of the purified enzyme.

    Science.gov (United States)

    Chang, Shu-Wei; Lee, Guan-Chiun; Shaw, Jei-Fu

    2006-08-09

    Candida rugosa lipase (CRL), an important industrial enzyme, possesses several different isoforms encoded by the high-identity lip gene family (lip1 to lip7). In this study, an additional N-terminal peptide in front of the lip3 gene was removed by PCR, and the 18 nonuniversal serine codons (CTG) of the lip3 gene were converted into universal serine codons (TCT) by means of an overlap extension PCR-based multiple-site-directed mutagenesis to express an active recombinant LIP3 in the yeast Pichia pastoris. The regional synthetic DNA fragment (339 bp) is first recombined by primer assembly with 20 overlapping nucleotides, followed by specific overlap extension PCR with outside primers containing restriction enzyme sites for directional cloning into the pGAPZalphaC vector. The results show that the production yield (0.687 unit/mL) of N-fused lip3 (nflip3) has an overall improvement of 69-fold relative to that (0.01 unit/mL) of lip3 and of 52-fold (0.47 unit/mL) of codon-optimized lip3 (colip3) relative to that (0.01 unit/mL) of non-codon-optimized lip3 (lip3), with the cultivation time set at 5 days. This finding demonstrates that the reservation of the N terminus and the regional codon optimization of the lip3 gene fragment at the 5' end can greatly increase the expression level of recombinant LIP3 in the P. pastoris system. The purified recombinant LIP3 shows distinct biochemical properties compared with other isoforms.

  6. Cloning, expression and alternative splicing of the novel isoform of hTCP11 gene

    DEFF Research Database (Denmark)

    Ma, Yong-xin; Zhang, Si-zhong; Wu, Qia-qing;

    2003-01-01

    To identify a novel isoform of hTCP11 gene and investigate its expression and alternative splicing.......To identify a novel isoform of hTCP11 gene and investigate its expression and alternative splicing....

  7. Isoforms of the Erythropoietin receptor in dopaminergic neurons of the Substantia Nigra.

    Science.gov (United States)

    Marcuzzi, Federica; Zucchelli, Silvia; Bertuzzi, Maria; Santoro, Claudio; Tell, Gianluca; Carninci, Piero; Gustincich, Stefano

    2016-11-01

    Erythropoietin receptor (EpoR) regulates erythrocytes differentiation in blood. In the brain, EpoR has been shown to protect several neuronal cell types from cell death, including the A9 dopaminergic neurons (DA) of the Substantia Nigra (SN). These cells form the nigrostriatal pathway and are devoted to the control of postural reflexes and voluntary movements. Selective degeneration of A9 DA neurons leads to Parkinson's disease. By the use of nanoCAGE, a technology that allows the identification of Transcription Start Sites (TSSs) at a genome-wide level, we have described the promoter-level expression atlas of mouse A9 DA neurons purified with Laser Capture Microdissection (LCM). Here, we identify mRNA variants of the Erythropoietin Receptor (DA-EpoR) transcribed from alternative TSSs. Experimental validation and full-length cDNA cloning is integrated with gene expression analysis in the FANTOM5 database. In DA neurons, the EpoR gene encodes for a N-terminal truncated receptor. Based on STAT5 phosphorylation assays, we show that the new variant of N-terminally truncated EpoR acts as decoy when co-expressed with the full-length form. A similar isoform is also found in human. This work highlights new complexities in the regulation of Erythropoietin (EPO) signaling in the brain.

  8. Separation of pegylated recombinant proteins and isoforms on CIM ion exchangers.

    Science.gov (United States)

    Gašperšič, Jernej; Podgornik, Aleš; Kramberger, Petra; Jarc, Marko; Jančar, Janez; Žorž, Mirjan; Krajnc, Nika Lendero

    2016-10-15

    Protein pegylation is a process of covalent attachment of a polyethylene glycol (PEG) group to the protein tertiary structure that can "mask" the agent from the immune system and also increases the hydrodynamic size of the agent. Usually the pegylation prolongs the protein stability in the organism due to reduced renal clearance and provides superior water solubility to hydrophobic molecules. The mono-pegylated form of protein is usually prefered for medical applications. Different conditions with different PEG reagents have to be tested to find optimal pegylation procedure with specific protein. The goal of this study was to prepare screening method for separation of random mono-pegylated protein. Cytochrome C and beta lactoglobulin were pegylated with four reagents and a complete screening of several chromatographic monoliths in ion exchange mode with different buffers was performed to optimaly separate each mono-pegylated protein. The screening method was developed that produces optimal separation of target pegylated protein on CIM monoliths. Because of short chromatographic run time, CIM monoliths are perfect candidates to test alot of parameters. The results obtained show that each protein has its own unique separation parameters (pH, ionexchange ligand, buffer type). Two biopharmaceuticals were isolated using protocol: super human leptin antagonist (SHLA) was purified from inclusion bodies and mono-pegylated super mouse leptin antagonist (SMLA) from pegylated mixture. During study it was observed that the convective interaction media (CIM) monoliths additionally discriminate between protein isoforms pegylated on different sites in 3D structure of the protein.

  9. Evidence that beta-hydroxyacyl-CoA dehydrase purified from rat liver microsomes is of peroxisomal origin.

    Science.gov (United States)

    Cook, L; Nagi, M N; Suneja, S K; Hand, A R; Cinti, D L

    1992-01-01

    The present study provides strong evidence that the previously isolated hepatic microsomal beta-hydroxyacyl-CoA dehydrase (EC 4.2.1.17), believed to be a component of the fatty acid chain-elongation system, is derived, not from the endoplasmic reticulum, but rather from the peroxisomes. The isolated dehydrase was purified over 3000-fold and showed optimal enzymic activity toward beta-hydroxyacyl-CoAs or trans-2-enoyl-CoAs with carbon chain lengths of 8-10. The purified preparation (VDH) displayed a pH optimum at 7.5 with beta-hydroxydecanoyl-CoA, and at 6.0 with beta-hydroxystearoyl-CoA. Competitive-inhibition studies suggested that VDH contained dehydrase isoforms, and SDS/PAGE showed three major bands at 47, 71 and 78 kDa, all of which reacted to antibody raised to the purified preparation. Immunocytochemical studies with anti-rabbit IgG to VDH unequivocally demonstrated gold particles randomly distributed throughout the peroxisomal matrix of liver sections from both untreated and di-(2-ethylhexyl) phthalate-treated rats. No labelling was associated with endoplasmic reticulum or with the microsomal fraction. Substrate-specificity studies and the use of antibodies to VDH and to the peroxisomal trifunctional protein indicated that VDH and the latter are separate enzymes. On the other hand, the VDH possesses biochemical characteristics similar to those of the D-beta-hydroxyacyl-CoA dehydrase recently isolated from rat liver peroxisomes [Li, Smeland & Schulz (1990) J. Biol. Chem. 265, 13629-13634; Hiltunen, Palosaari & Kunau (1989) J. Biol. Chem. 264, 13536-13540]. Neither enzyme utilizes crotonoyl-CoA or cis-2-enoyl-CoA as substrates, but both enzymes convert trans-2-enoyl substrates into the D-isomer only. In addition, the VDH also contained beta-oxoacyl-CoA reductase (beta-hydroxyacyl-CoA dehydrogenase) activity, which co-purified with the dehydrase. Images Fig. 4. Fig. 7. Fig. 8. PMID:1417796

  10. Mast cells express novel functional IL-15 receptor alpha isoforms.

    Science.gov (United States)

    Bulanova, Elena; Budagian, Vadim; Orinska, Zane; Krause, Hans; Paus, Ralf; Bulfone-Paus, Silvia

    2003-05-15

    Mast cells previously have been reported to be regulated by IL-15 and to express a distinct IL-15R, termed IL-15RX. To further examine IL-15 binding and signaling in mast cells, we have studied the nature of the IL-15R and some of its biological activities in these cells. In this study, we report the existence of three novel isoforms of the IL-15R alpha chain in murine bone marrow-derived mast cells as a result of an alternative exon-splicing mechanism within the IL-15R alpha gene. These correspond to new mRNA transcripts lacking exon 4; exons 3 and 4; or exons 3, 4, and 5 (IL-15R alpha Delta 4, IL-15R alpha Delta 3,4, IL-15R alpha Delta 3,4,5). After transient transfection in COS-7 cells, all IL-15R alpha isoforms associate with the Golgi apparatus, the endoplasmic reticulum, the perinuclear space, and the cell membrane. Analysis of glycosylation pattern demonstrates the usage of a single N-glycosylation site, while no O-glycosylation is observed. Importantly, IL-15 binds with high affinity to, and promotes the survival of, murine BA/F3 cells stably transfected with the IL-15R alpha isoforms. Furthermore, we report that signaling mediated by IL-15 binding to the newly identified IL-15R alpha isoforms involves the phosphorylation of STAT3, STAT5, STAT6, Janus kinase 2, and Syk kinase. Taken together, our data indicate that murine mast cells express novel, fully functional IL-15R alpha isoforms, which can explain the selective regulatory effects of IL-15 on these cells.

  11. Growth hormone isoforms in a girl with gigantism.

    Science.gov (United States)

    Ng, L L; Chasalow, F I; Escobar, O; Blethen, S L

    1999-01-01

    Several previous investigations have suggested that there may be different growth hormone isoforms in patients with acromegaly. We used three different site-specific monoclonal antibodies (MAbs) to investigate growth hormone (GH) isoforms in serum from an 8 year-old girl with a GH and prolactin secreting adenoma. The pattern of GH-immunoreactivity was dependent on the circumstances of collection. Serum obtained after oral glucose had very little cross reactivity with MAb 352 although concentrations of up to 15 micrograms/l were found with two other MAbs, 033 and 665. MAb 352 does not recognize the 20,000 dalton isoform of GH (20K) while both MAb 033 and 665 do. The same pattern of GH immunoreactivity (low MAb 352, equal and higher MAb 033 and 665) was seen in other baseline samples. In contrast, samples obtained after TRH/GnRH showed immunoreactivity patterns expected for a mixture of 22,000 dalton isoform of GH (22K) with only a small amount of 20K. GH samples obtained during sleep showed both patterns with episodic peaks with equal immunoreactivity superimposed on the basal pattern (decreased activity with MAb 352). Affinity chromatography of basal samples showed that a portion of the GH immunoreactivity was neither 22K nor 20K, although in stimulated samples, over 70% of GH was 22K or 20K GH. In conclusion, the nature of GH isoforms present in serum varies with GH concentration. These differences may contribute to the known difficulty in correlating disease activity and random GH measurements in patients with GH secreting adenomas.

  12. Identification of cardiac myofilament protein isoforms using multiple mass spectrometry based approaches

    NARCIS (Netherlands)

    Kooij, V.; Venkatraman, V.; Kirk, J.A.; Ubaida-Mohien, C.; Graham, D.R.; Faber, M.J.; Eyk, J.E. Van

    2014-01-01

    PURPOSE: The identification of protein isoforms in complex biological samples is challenging. We, therefore, used an MS approach to unambiguously identify cardiac myofilament protein isoforms based on the observation of a tryptic peptide consisting of a sequence unique to a particular isoform. EXPER

  13. Expression, purification and enzymatic characterization of the catalytic domains of human tryptophan hydroxylase isoforms

    DEFF Research Database (Denmark)

    Windahl, Michael Skovbo; Boesen, Jane; Karlsen, Pernille Efferbach;

    2009-01-01

    Tryptophan hydroxylase exists in two isoforms: Isoform 1 catalyses the first and rate-limiting step in the synthesis of serotonin in the peripheral parts of the body while isoform 2 catalyses this step in the brain. The catalytic domains of human tryptophan hydroxylase 1 and 2 have been expressed...

  14. Ferromagnetic Levan Composite: An Affinity Matrix to Purify Lectin

    Directory of Open Access Journals (Sweden)

    Renata Angeli

    2009-01-01

    Full Text Available A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A and Cratylia mollis (Cramoll 1 and Cramoll 1,4 did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammonium sulfate fractionation, affinity chromatography on Sephadex G-75, and ion exchange chromatography through a CM-cellulose column.

  15. Identification of two metallo- thionein isoforms by molecu-lar cloning of their cDNAs infresh-water fish, crucian carp(Carassius-cuvieri)

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Generally, there are two major isoforms of me- tallothionein (MT)in mammals. In this study two cDNAs of metallothionein, MT-A and MT-B, in a fresh-water fish, crucian carp (Carassius cuvieri), were cloned by the rapid amplification of cDNA ends (RACE). The homology of their reading frame is about 92.3%. The sequence analysis of both cDNAs gave the structures of coding regions corresponding to 60 amino acid residues, and the structures of complete 3′-untranslated regions in which a significant difference in the size of their 3′-untranslated regions (130 bp for MT-A and 280 bp for MT-B) exists. The results of amino acid sequenc-ing of both MT-1 and MT-2 purified by HPLC are identical to those deduced from MT cDNA genes, indicating that MT-1 is from MT-A gene and MT-2 is from MT-B gene respec-tively. No blocking in the N-terminal of MT-2 isoform was the first case found in vertebrates, most of which were block-ed by acetylation. These results suggest that there were dif-ferential controls at the transcription level and after transla-tion of these two MT isoforms. And this gives a clue to un-derstand the diversities of their functions.

  16. The effects of salt stress cause a diversion of basal metabolism in barley roots: possible different roles for glucose-6-phosphate dehydrogenase isoforms.

    Science.gov (United States)

    Cardi, Manuela; Castiglia, Daniela; Ferrara, Myriam; Guerriero, Gea; Chiurazzi, Maurizio; Esposito, Sergio

    2015-01-01

    In this study the effects of salt stress and nitrogen assimilation have been investigated in roots of hydroponically-grown barley plants exposed to 150 mM NaCl, in presence or absence of ammonium as the sole nitrogen source. Salt stress determines a diversion of root metabolism towards the synthesis of osmolytes, such as glycine betaine and proline, and increased levels of reduced glutathione. The metabolic changes triggered by salt stress result in a decrease in both activities and protein abundance of key enzymes, namely GOGAT and PEP carboxylase, and in a slight increase in HSP70. These variations would enhance the requirement for reductants supplied by the OPPP, consistently with the observed increase in total G6PDH activity. The involvement and occurrence of the different G6PDH isoforms have been investigated, and the kinetic properties of partially purified cytosolic and plastidial G6PDHs determined. Bioinformatic analyses examining co-expression profiles of G6PDHs in Arabidopsis and barley corroborate the data presented. Moreover, the gene coding for the root P2-G6PDH isoform was fully sequenced; the biochemical properties of the corresponding protein were examined experimentally. The results are discussed in the light of the possible distinct roles and regulation of the different G6PDH isoforms during salt stress in barley roots.

  17. Hyperglycemic activity of the recombinant crustacean hyperglycemic hormone B1 isoform (CHH-B1) of the Pacific white shrimp Litopenaeus vannamei.

    Science.gov (United States)

    Camacho-Jiménez, Laura; Sánchez-Castrejón, Edna; Ponce-Rivas, Elizabeth; Muñoz-Márquez, Ma Enriqueta; Aguilar, Manuel B; Re, Ana Denisse; Díaz, Fernando

    2015-09-01

    Crustacean hyperglycemic hormone (CHH) is the most abundant neuropeptide produced by the X-organ/sinus gland (XO/SG) complex in the crustacean eyestalk. CHH plays a principal role in the control of glucose metabolism. The CHH-B1 isoform is produced in the eyestalk of Litopenaeus vannamei by alternative splicing of the chhB gene and its cDNA sequence has revealed that this isoform has a non-amidated C-terminal residue (CHH-like peptide). In this work, a recombinant CHH-B1 (rCHH-B1) with a sequence identical to the native hormone was expressed in the methylotrophic yeast Pichia pastoris X-33 and purified from the culture medium by RP-HPLC. The identity of the purified rCHH-B1 was confirmed by N-terminal sequencing and by using an anti-CHH-B1 polyclonal antibody. An in vivo assay showed that the hyperglycemic effect was dependant of the dosage of rCHH-B1, and the maximal hyperglycemic response was obtained with 250pmol treatment. These results suggest that the amino acid sequence of the C-terminus and its correct structure are both important for the hyperglycemic activity of naturally occurring non-amidated CHH peptides, such as CHH-B1. CHH-B1 appears to be the first reported CHH-like peptide with significant hyperglycemic activity produced in the sinus gland of a penaeid shrimp.

  18. EGFR soluble isoforms and their transcripts are expressed in meningiomas.

    Science.gov (United States)

    Guillaudeau, Angélique; Durand, Karine; Bessette, Barbara; Chaunavel, Alain; Pommepuy, Isabelle; Projetti, Fabrice; Robert, Sandrine; Caire, François; Rabinovitch-Chable, Hélène; Labrousse, François

    2012-01-01

    The EGFR (epidermal growth factor receptor) is involved in the oncogenesis of many tumors. In addition to the full-length EGFR (isoform a), normal and tumor cells produce soluble EGFR isoforms (sEGFR) that lack the intracellular domain. sEGFR isoforms b, c and d are encoded by EGFR variants 2 (v2), 3 (v3) and 4 (v4) mRNA resulting from gene alternative splicing. Accordingly, the results of EGFR protein expression analysis depend on the domain targeted by the antibodies. In meningiomas, EGFR expression investigations mainly focused on EGFR isoform a. sEGFR and EGFRvIII mutant, that encodes a constitutively active truncated receptor, have not been studied. In a 69 meningiomas series, protein expression was analyzed by immunohistochemistry using extracellular domain targeted antibody (ECD-Ab) and intracellular domain targeted antibody (ICD-Ab). EGFRv1 to v4 and EGFRvIII mRNAs were quantified by RT-PCR and EGFR amplification revealed by MLPA. Results were analyzed with respect to clinical data, tumor resection (Simpson grade), histological type, tumor grade, and patient outcome.Immunochemical staining was stronger with ECD-Ab than with ICD-Ab. Meningiomas expressed EGFRv1 to -v4 mRNAs but not EGFRvIII mutant. Intermediate or high ECD-Ab staining and high EGFRv1 to v4 mRNA levels were associated to a better progression free survival (PFS). PFS was also improved in women, when tumor resection was evaluated as Simpson 1 or 2, in grade I vs. grade II and III meningiomas and when Ki67 labeling index was lower than 10%. Our results suggest that, EGFR protein isoforms without ICD and their corresponding mRNA variants are expressed in meningiomas in addition to the whole isoform a. EGFRvIII was not expressed. High expression levels seem to be related to a better prognosis. These results indicate that the oncogenetic mechanisms involving the EGFR pathway in meningiomas could be different from other tumor types.

  19. EGFR soluble isoforms and their transcripts are expressed in meningiomas.

    Directory of Open Access Journals (Sweden)

    Angélique Guillaudeau

    Full Text Available The EGFR (epidermal growth factor receptor is involved in the oncogenesis of many tumors. In addition to the full-length EGFR (isoform a, normal and tumor cells produce soluble EGFR isoforms (sEGFR that lack the intracellular domain. sEGFR isoforms b, c and d are encoded by EGFR variants 2 (v2, 3 (v3 and 4 (v4 mRNA resulting from gene alternative splicing. Accordingly, the results of EGFR protein expression analysis depend on the domain targeted by the antibodies. In meningiomas, EGFR expression investigations mainly focused on EGFR isoform a. sEGFR and EGFRvIII mutant, that encodes a constitutively active truncated receptor, have not been studied. In a 69 meningiomas series, protein expression was analyzed by immunohistochemistry using extracellular domain targeted antibody (ECD-Ab and intracellular domain targeted antibody (ICD-Ab. EGFRv1 to v4 and EGFRvIII mRNAs were quantified by RT-PCR and EGFR amplification revealed by MLPA. Results were analyzed with respect to clinical data, tumor resection (Simpson grade, histological type, tumor grade, and patient outcome.Immunochemical staining was stronger with ECD-Ab than with ICD-Ab. Meningiomas expressed EGFRv1 to -v4 mRNAs but not EGFRvIII mutant. Intermediate or high ECD-Ab staining and high EGFRv1 to v4 mRNA levels were associated to a better progression free survival (PFS. PFS was also improved in women, when tumor resection was evaluated as Simpson 1 or 2, in grade I vs. grade II and III meningiomas and when Ki67 labeling index was lower than 10%. Our results suggest that, EGFR protein isoforms without ICD and their corresponding mRNA variants are expressed in meningiomas in addition to the whole isoform a. EGFRvIII was not expressed. High expression levels seem to be related to a better prognosis. These results indicate that the oncogenetic mechanisms involving the EGFR pathway in meningiomas could be different from other tumor types.

  20. Creating and purifying an observation instrument using the generalizability theory

    Directory of Open Access Journals (Sweden)

    Elena Rodríguez-Naveiras

    2013-12-01

    Full Text Available The control of quality of data it is one of the most relevant aspects in observational researches. The Generalizability Theory (GT provides a method of analysis that allows us to isolate the various sources of error measurement. At the same time, it helps us to determine the extent to which various factors can change and analyze the effect on the generalizability coefficient. In the work shown here, there are two studies aimed to creating and purifying an observation instrument, Observation Protocol in the Teaching Functions (Protocolo de Funciones Docentes, PROFUNDO, v1 and v2, for behavioral assessment which has been carried out by instructors in a social-affective out-of-school program. The reliability and homogeneity studies are carried out once the instrument has been created and purified. The reliability study will be done through the GT method taking both codes (c and agents (a as differential facets in. The generalization will be done through observers using a crossed multi-faceted design (A × O × C. In the homogeneity study the generalization facet will be done through codes using the same design that the reliability study.

  1. Synthesis of gold and silver nanoparticles using purified URAK.

    Science.gov (United States)

    Deepak, Venkataraman; Umamaheshwaran, Paneer Selvam; Guhan, Kandasamy; Nanthini, Raja Amrisa; Krithiga, Bhaskar; Jaithoon, Nagoor Meeran Hasika; Gurunathan, Sangiliyandi

    2011-09-01

    This study aims at developing a new eco-friendly process for the synthesis of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) using purified URAK. URAK is a fibrinolytic enzyme produced by Bacillus cereus NK1. The enzyme was purified and used for the synthesis of AuNPs and AgNPs. The enzyme produced AgNPs when incubated with 1 mM AgNO3 for 24 h and AuNPs when incubated with 1 mM HAuCl4 for 60 h. But when NaOH was added, the synthesis was rapid and occurred within 5 min for AgNPs and 12 h for AuNPs. The synthesized nanoparticles were characterized by a peak at 440 nm and 550 nm in the UV-visible spectrum. TEM analysis showed that AgNPs of the size 60 nm and AuNPs of size 20 nm were synthesized. XRD confirmed the crystalline nature of the nanoparticles and AFM showed the morphology of the nanoparticle to be spherical. FT-IR showed that protein was responsible for the synthesis of the nanoparticles. This process is highly simple, versatile and produces AgNPs and AuNPs in environmental friendly manner. Moreover, the synthesized nanoparticles were found to contain immobilized enzyme. Also, URAK was tested on RAW 264.7 macrophage cell line and was found to be non-cytotoxic until 100 μg/ml.

  2. A Purified Recombinant Lipopeptide as Adjuvant for Cancer Immunotherapy

    Directory of Open Access Journals (Sweden)

    Ying-Chyi Song

    2014-01-01

    Full Text Available Synthetic lipopeptides have been widely used as vaccine adjuvants to enhance immune responses. The present study demonstrated that the tryptic N-terminal fragment of the lipoprotein rlipo-D1E3 (lipo-Nter induces superior antitumor effects compared to a synthetic lipopeptide. The lipo-Nter was purified and formulated with protein or peptide vaccines to determine if lipo-Nter could be used as a novel adjuvant and could induce antitumor immunity in a cervical cancer model. Purified lipo-Nter activated the maturation of bone marrow-derived dendritic cells (BM-DCs, leading to the secretion of TNF-α through TLR2/6 but not TLR1/2. A recombinant mutant HPV16 E7 (rE7m protein was mixed with lipo-Nter to immunize the mice; the anti-E7 antibody titers were increased, and the T helper cells were skewed toward the Th1 fate (increased IL-2 and decreased IL-5 secretion. Single-dose injection of rE7m and lipo-Nter inhibited tumor growth, but the injection of rE7m alone did not. Accordingly, lipo-Nter also enhanced the antitumor immunity of the E7-derived peptide but not the synthetic lipopeptide (Pam3CSK4. We demonstrated that the lipo-Nter of a bacterial-derived recombinant lipoprotein is a novel adjuvant that could be used for the development of a new generation of vaccines.

  3. Life cycle assessment comparison of photocatalytic coating and air purifier.

    Science.gov (United States)

    Tichá, Marie; Žilka, Miroslav; Stieberová, Barbora; Freiberg, František

    2016-07-01

    This article presents a comparison of 2 very different options for removal of undesirable microorganisms and airborne pollutants from the indoor environment of hospitals, schools, homes, and other enclosed spaces using air purifiers and photocatalytic coatings based on nano titanium dioxide (TiO2 ). Both products were assessed by life cycle assessment (LCA) methodology from cradle-to-grave. The assessment also includes comparison of 2 different nano TiO2 production technologies, one by continuous hydrothermal synthesis and the other by a sulfate process. Results of the study showed a relatively large contribution of photocatalytic coatings to reducing the effects of selected indices in comparison with an air purifier, regardless of which nano TiO2 production method is used. Although the impacts of the sulfate process are significantly lower compared to those of hydrothermal synthesis when viewed in terms of production alone, taken in the context of the entire product life cycle, the net difference becomes less significant. The study has been elaborated within the Sustainable Hydrothermal Manufacturing of Nanomaterials (SHYMAN) project, which aims to develop competitive and sustainable continuous nanoparticle (NP) production technology based on supercritical hydrothermal synthesis. Integr Environ Assess Manag 2016;12:478-485. © 2016 SETAC.

  4. Production and Immobilization of Partially Purified Lipase From Penicillium chrysogenum

    Directory of Open Access Journals (Sweden)

    Shafei, M. S.

    2010-01-01

    Full Text Available An extracellular lipase from Penicillium chrysogenum produced maximal activity 225 U/mL after four days at pH 6.5. It was partially purified 4.1 fold by ammonium sulphate precipitation (70%. The enzyme was immobilized on various carriers viz. alginate, k-carrageenan and polyacrylamide gel. The immobilization yield of enzyme immobilized in kcarrageenan and polyacrylamide gel (63.41% and 48.93% respectively was low in comparison to that immobilized with alginate (81.57%. Different concentrations of alginate were tried to study their effect on lipase production. Maximum immobilization yield was observed with 3% alginate. The optimal pH of the partially purified lipase was 7.5 and the optimum temperature was 35 °C. At 60 °C the immobilized enzyme retained 62.79% of its activity. Broader pH tolerance and higher heat stability could be achieved by this method. Immobilized lipase retained 72.09% relative activity after six hydrolysis cycles.

  5. Tea derived galloylated polyphenols cross-link purified gastrointestinal mucins.

    Directory of Open Access Journals (Sweden)

    Pantelis Georgiades

    Full Text Available Polyphenols derived from tea are thought to be important for human health. We show using a combination of particle tracking microrheology and small-angle neutron scattering that polyphenols acts as cross-linkers for purified gastrointestinal mucin, derived from the stomach and the duodenum. Both naturally derived purified polyphenols, and green and black tea extracts are shown to act as cross-linkers. The main active cross-linking component is found to be the galloylated forms of catechins. The viscosity, elasticity and relaxation time of the mucin solutions experience an order of magnitude change in value upon addition of the polyphenol cross-linkers. Similarly small-angle neutron scattering experiments demonstrate a sol-gel transition with the addition of polyphenols, with a large increase in the scattering at low angles, which is attributed to the formation of large scale (>10 nm heterogeneities during gelation. Cross-linking of mucins by polyphenols is thus expected to have an impact on the physicochemical environment of both the stomach and duodenum; polyphenols are expected to modulate the barrier properties of mucus, nutrient absorption through mucus and the viscoelastic microenvironments of intestinal bacteria.

  6. Magnetism for understanding catalyst analysis of purified carbon nanotubes

    Science.gov (United States)

    Bellouard, Christine; Mercier, Guillaume; Cahen, Sébastien; Ghanbaja, Jaafar; Medjahdi, Ghouti; Gleize, Jérôme; Lamura, Gianrico; Hérold, Claire; Vigolo, Brigitte

    2016-08-01

    The precise quantification of catalyst residues in purified carbon nanotubes is often a major issue in view of any fundamental and/or applicative studies. More importantly, since the best CNTs are successfully grown with magnetic catalysts, their quantification becomes strictly necessary to better understand intrinsic properties of CNT. For these reasons, we have deeply analyzed the catalyst content remained in nickel-yttrium arc-discharge single walled carbon nanotubes purified by both a chlorine-gas phase and a standard acid-based treatment. The study focuses on Ni analysis which has been investigated by transmission electron microscopy, X-ray diffraction, thermogravimetry analysis, and magnetic measurements. In the case of the acid-based treatment, all quantifications result in a decrease of the nanocrystallized Ni by a factor of two. In the case of the halogen gas treatment, analysis and quantification of Ni content is less straightforward: a huge difference appears between X-ray diffraction and thermogravimetry results. Thanks to magnetic measurements, this disagreement is explained by the presence of Ni2+ ions, belonging to NiCl2 formed during the Cl-based purification process. In particular, NiCl2 compound appears under different magnetic/crystalline phases: paramagnetic or diamagnetic, or well intercalated in between carbon sheets with an ordered magnetic phase at low temperature.

  7. Proteomic Validation of Transcript Isoforms, Including Those Assembled from RNA-Seq Data

    DEFF Research Database (Denmark)

    Tay, Aidan P; Pang, Chi Nam Ignatius; Twine, Natalie a;

    2015-01-01

    data, and proteomic analysis of the same sample, can identify protein isoforms. RNA-seq data from human mesenchymal (hMSC) stem cells were analyzed with our new TranscriptCoder tool to generate a database of protein isoform sequences. MS/MS data from matching hMSC samples were then matched against...... the TranscriptCoder-derived database, along with Ensembl and the neXtProt database. Querying the TranscriptCoder-derived or Ensembl database could unambiguously identify ∼450 protein isoforms, with isoform-specific proteotypic peptides, including candidate hMSC-specific isoforms for the genes DPYSL2 and FXR1...

  8. Characterization of four hemocyanin isoforms in Litopenaeus vannamei

    Institute of Scientific and Technical Information of China (English)

    XU Jingxiang; RUAN Lingwei; LI Zhen; YU Xiaoman; LI Sedong; SHI Hong; XU Xun

    2015-01-01

    In this study, the gene encoding hemocyanin subunit L, LvHcL, was cloned from Litopenaeus vannamei and the genomic organization was characterized. This gene was diverse with many SNPs and also had at least four isoforms, while one of them (LvHcL4) only had two exons and the exon2 was missed. Transcription analysis showed that these isoforms of LvHcL were up-regulated after WSSV challenge in WSSV-resistant shrimp, while the transcriptions were decreased constantly in WSSV-susceptible shrimp. It is suggested that the hemocyanin had rich polymorphism and was involved in the antiviral response. These results could extend our previous findings and provide insights into the immune feature of hemocyanin, which would be helpful for further studies aimed at antiviral mechanism in inver-tebrate.

  9. Disulfide isoforms of recombinant glia maturation factor beta.

    Science.gov (United States)

    Zaheer, A; Lim, R

    1990-09-14

    Recombinant human glia maturation factor beta (r-hGMF-beta) is a single-chain polypeptide (141 amino acid residues) containing three cysteines, at positions 7, 86 and 95. Nascent r-hGMF-beta exists in the reduced state and has no biological activity. The protein can be activated through oxidative refolding by incubation with a mixture of reduced and oxidized glutathione. Reverse-phase HPLC analysis of the refolded r-hGMF-beta shows the presence of four peaks, corresponding to the reduced form plus three newly generated intrachain disulfide-containing isoforms predicted from the number of cysteine residues. Only one isoform shows biological activity when tested for growth suppression on C6 glioma cells. We infer from the HPLC elution pattern that the active form contains the disulfide bridge Cys86-Cys95.

  10. First measurement of the polarisation asymmetry of a gamma-ray beam between 1.7 to 74 MeV with the HARPO TPC

    Science.gov (United States)

    Gros, P.; Amano, S.; Attié, D.; Bernard, D.; Bruel, P.; Calvet, D.; Colas, P.; Daté, S.; Delbart, A.; Frotin, M.; Geerebaert, Y.; Giebels, B.; Götz, D.; Hashimoto, S.; Horan, D.; Kotaka, T.; Louzir, Marc; Minamiyama, Y.; Miyamoto, S.; Ohkuma, H.; Poilleux, Patrick; Semeniouk, I.; Sizun, P.; Takemoto, A.; Yamaguchi, M.; Wang, S.

    2016-07-01

    Current γ-ray telescopes suffer from a gap in sensitivity in the energy range between 100 keV and 100 MeV, and no polarisation measurement has ever been done on cosmic sources above 1 MeV. Past and present e+e- pair telescopes are limited at lower energies by the multiple scattering of electrons in passive tungsten converter plates. This results in low angular resolution, and, consequently, a drop in sensitivity to point sources below 1 GeV. The polarisation information, which is carried by the azimuthal angle of the conversion plane, is lost for the same reasons. HARPO is an R&D program to characterise the operation of a gaseous detector (a Time Projection Chamber or TPC) as a high angular-resolution and sensitivity telescope and polarimeter for γ-rays from cosmic sources. It represents a first step towards a future space instrument in the MeV-GeV range. We built and characterised a 30cm cubic demonstrator [SPIE 91441M], and put it in a polarised γ-ray beam at the NewSUBARU accelerator in Japan. Data were taken at photon energies from 1.74MeV to 74MeV and with different polarisation configurations. We describe the experimental setup in beam. We then describe the software we developed to reconstruct the photon conversion events, with special focus on low energies. We also describe the thorough simulation of the detector used to compare results. Finally we will present the performance of the detector as extracted from this analysis and preliminary measurements of the polarisation asymmetry. This beam-test qualification of a gas TPC prototype in a γ-ray beam could open the way to high-performance -ray astronomy and polarimetry in the MeV-GeV energy range in the near future.

  11. Accurate γ and MeV-electron track reconstruction with an ultra-low diffusion Xenon/TMA TPC at 10 atm

    Science.gov (United States)

    González-Díaz, Diego; Álvarez, V.; Borges, F. I. G.; Camargo, M.; Cárcel, S.; Cebrián, S.; Cervera, A.; Conde, C. A. N.; Dafni, T.; Díaz, J.; Esteve, R.; Fernandes, L. M. P.; Ferrario, P.; Ferreira, A. L.; Freitas, E. D. C.; Gehman, V. M.; Goldschmidt, A.; Gómez-Cadenas, J. J.; Gutiérrez, R. M.; Hauptman, J.; Hernando Morata, J. A.; Herrera, D. C.; Irastorza, I. G.; Labarga, L.; Laing, A.; Liubarsky, I.; Lopez-March, N.; Lorca, D.; Losada, M.; Luzón, G.; Marí, A.; Martín-Albo, J.; Martínez-Lema, G.; Martínez, A.; Miller, T.; Monrabal, F.; Monserrate, M.; Monteiro, C. M. B.; Mora, F. J.; Moutinho, L. M.; Muñoz Vidal, J.; Nebot-Guinot, M.; Nygren, D.; Oliveira, C. A. B.; Pérez, J.; Pérez Aparicio, J. L.; Querol, M.; Renner, J.; Ripoll, L.; Rodríguez, J.; Santos, F. P.; dos Santos, J. M. F.; Serra, L.; Shuman, D.; Simón, A.; Sofka, C.; Sorel, M.; Toledo, J. F.; Torrent, J.; Tsamalaidze, Z.; Veloso, J. F. C. A.; Villar, J. A.; Webb, R.; White, J. T.; Yahlali, N.; Azevedo, C.; Aznar, F.; Calvet, D.; Castel, J.; Ferrer-Ribas, E.; García, J. A.; Giomataris, I.; Gómez, H.; Iguaz, F. J.; Lagraba, A.; Le Coguie, A.; Mols, J. P.; Şahin, Ö.; Rodríguez, A.; Ruiz-Choliz, E.; Segui, L.; Tomás, A.; Veenhof, R.

    2015-12-01

    We report the performance of a 10 atm Xenon/trimethylamine time projection chamber (TPC) for the detection of X-rays (30 keV) and γ-rays (0.511-1.275 MeV) in conjunction with the accurate tracking of the associated electrons. When operated at such a high pressure and in 1%-admixtures, trimethylamine (TMA) endows Xenon with an extremely low electron diffusion (1.3 ± 0.13 mm - σ (longitudinal), 0.95 ± 0.20 mm - σ (transverse) along 1 m drift) besides forming a convenient 'Penning-Fluorescent' mixture. The TPC, that houses 1.1 kg of gas in its fiducial volume, operated continuously for 100 live-days in charge amplification mode. The readout was performed through the recently introduced microbulk Micromegas technology and the AFTER chip, providing a 3D voxelization of 8 mm × 8 mm × 1.2 mm for approximately 10 cm/MeV-long electron tracks. Resolution in energy (ε) at full width half maximum (R) inside the fiducial volume ranged from R = 14.6 % (30 keV) to R = 4.6 %(1.275 MeV). This work was developed as part of the R&D program of the NEXT collaboration for future detector upgrades in the search of the neutrino-less double beta decay (ββ 0 ν) in 136Xe, specifically those based on novel gas mixtures. Therefore we ultimately focus on the calorimetric and topological properties of the reconstructed MeV-electron tracks. In particular, the obtained energy resolution has been decomposed in its various contributions and improvements towards achieving the R = 1.4 %√{ 1 MeV / ε } levels obtained in small sensors are discussed.

  12. Method for analysing glycoprotein isoforms by capillary electrophoresis

    OpenAIRE

    Frutos, Mercedes de; Díez-Masa, José Carlos; Morales-Cid, Gabriel

    2011-01-01

    [EN] The present invention relates to a new method for the purification, concentration, separation and determination of the isoforms of alpha-1-acid glycoprotein (AGP) in human blood serum samples using capillary electrophoresis. The new method is based on the immunocapture and preconcentration of the sample within the separation capillary by using an immunoadsorbent phase magnetically immobilized within the electrophoresis capillary and the subsequent desorption and separation of the glycopr...

  13. Functional differences between L- and T-plastin isoforms.

    Science.gov (United States)

    Arpin, M; Friederich, E; Algrain, M; Vernel, F; Louvard, D

    1994-12-01

    Fimbrins/plastins are a family of highly conserved actin-bundling proteins. They are present in all eukaryotic cells including yeast, but each isoform displays a remarkable tissue specificity. T-plastin is normally found in epithelial and mesenchymal cells while L-plastin is present in hematopoietic cells. However, L-plastin has been also found in tumor cells of non-hematopoietic origin (Lin, C.-S., R. H. Aebersold, S. B. Kent, M. Varma, and J. Leavitt. 1988. Mol. Cell. Biol. 8:4659-4668; Lin, C.-S., R. H. Aebersold, and J. Leavitt. 1990. Mol. Cell. Biol. 10: 1818-1821). To learn more about the biological significance of their tissue specificity, we have overproduced the T- and L-plastin isoforms in a fibroblast-like cell line, CV-1, and in a polarized epithelial cell line, LLC-PK1. In CV-1 cells, overproduction of T- and L-plastins induces cell rounding and a concomitant reorganization of actin stress fibers into geodesic structures. L-plastin remains associated with microfilaments while T-plastin is almost completely extracted after treatment of the cells with non-ionic detergent. In LLC-PK1 cells, T-plastin induces shape changes in microvilli and remains associated with microvillar actin filaments after detergent extraction while L-plastin has no effect on these structures and is completely extracted. The effect of T-plastin on the organization of microvilli differs from that of villin, another actin-bundling protein. Our experiments indicate that these two isoforms play differing roles in actin filament organization, and do so in a cell type-specific fashion. Thus it is likely that these plastin isoforms play fundamentally different roles in cell function.

  14. [Clinical relevance of myosin isoforms in the diaphragm].

    Science.gov (United States)

    Gayan-Ramirez, G; Decramer, M

    2000-06-01

    The diaphragm as a striated muscle is characterized by the repetition of a single element arranged in series: the sarcomere containing two kinds of myofilaments: a thick one constituted by the myosin, and a thin one primarily composed of actin. The myosin molecule consists of two heads where two myosin heavy chains (MHC) are fixed, a flexible hinge with two light (MLC) chains, and long rod-shaped tails. The diaphragm contains 4 MHC isoforms (MHC-slow, MHC-2A, MHC-2B, MHC-2X) and 6 MLC isoforms (MLC-1f, MLC-3f, MLC-1sa, MLC-1sb, MLC-2f, MLC-2s/v). In humans, the diaphragm contains mainly fibers expressing the isoforms MHC-slow, MHC-2A, and MLC-2f, MLC-2s et MLC-1f. For the mechanical properties of the different isoforms, there is a gradient from the MHC-slow to the MHC-2A, MHC-2B and MHC-2X/2B. According to the circumstances, the diaphragm will adapt towards a slow profile (COPD, cardiac failure and in animals: Duchenne muscular dystrophy, denervation-1 week, age-female, corticosteroids, chronic stimulation), or a fast profile (in animals: chronic hypoxia, denervation-2 weeks, age-males) or a more oxidative profile (in animals: cachexia, obesity). The reasons why the diaphragm adapts towards a slower or a faster muscle are not known. In fact, for a given pathological situation, several factors are able to influence the fiber composition of the diaphragm. Therefore, the net result of the influence of these different factors in terms of MHC and MLC diaphragm adaptation is difficult to predict.

  15. Regulation of NADPH oxidase 5 by protein kinase C isoforms.

    Directory of Open Access Journals (Sweden)

    Feng Chen

    Full Text Available NADPH oxidase5 (Nox5 is a novel Nox isoform which has recently been recognized as having important roles in the pathogenesis of coronary artery disease, acute myocardial infarction, fetal ventricular septal defect and cancer. The activity of Nox5 and production of reactive oxygen species is regulated by intracellular calcium levels and phosphorylation. However, the kinases that phosphorylate Nox5 remain poorly understood. Previous studies have shown that the phosphorylation of Nox5 is PKC dependent, but this contention was based on the use of pharmacological inhibitors and the isoforms of PKC involved remain unknown. Thus, the major goals of this study were to determine whether PKC can directly regulate Nox5 phosphorylation and activity, to identify which isoforms are involved in the process, and to understand the functional significance of this pathway in disease. We found that a relatively specific PKCα inhibitor, Ro-32-0432, dose-dependently inhibited PMA-induced superoxide production from Nox5. PMA-stimulated Nox5 activity was significantly reduced in cells with genetic silencing of PKCα and PKCε, enhanced by loss of PKCδ and the silencing of PKCθ expression was without effect. A constitutively active form of PKCα robustly increased basal and PMA-stimulated Nox5 activity and promoted the phosphorylation of Nox5 on Ser490, Thr494, and Ser498. In contrast, constitutively active PKCε potently inhibited both basal and PMA-dependent Nox5 activity. Co-IP and in vitro kinase assay experiments demonstrated that PKCα directly binds to Nox5 and modifies Nox5 phosphorylation and activity. Exposure of endothelial cells to high glucose significantly increased PKCα activation, and enhanced Nox5 derived superoxide in a manner that was in prevented by a PKCα inhibitor, Go 6976. In summary, our study reveals that PKCα is the primary isoform mediating the activation of Nox5 and this maybe of significance in our understanding of the vascular

  16. Isoform expression in the multiple soluble malate dehydrogenase of Hoplias malabaricus (Erythrinidae, Characiformes

    Directory of Open Access Journals (Sweden)

    M. R. Aquino-Silva

    Full Text Available Kinetic properties and thermal stabilities of Hoplias malabaricus liver and skeletal muscle unfractionated malate dehydrogenase (MDH, EC 1.1.1.37 and its isolated isoforms were analyzed to further study the possible sMDH-A* locus duplication evolved from a recent tandem duplication. Both A (A1 and A2 and B isoforms had similar optima pH (7.5-8.0. While Hoplias A isoform could not be characterized as thermostable, B could as thermolabile. A isoforms differed from B isoform in having higher Km values for oxaloacetate. The possibly duplicated A2 isoform showed higher substrate affinity than the A1. Hoplias duplicated A isoforms may influence the direction of carbon flow between glycolisis and gluconeogenesis.

  17. Isoform expression in the multiple soluble malate dehydrogenase of Hoplias malabaricus (Erythrinidae, Characiformes

    Directory of Open Access Journals (Sweden)

    Aquino-Silva M. R.

    2003-01-01

    Full Text Available Kinetic properties and thermal stabilities of Hoplias malabaricus liver and skeletal muscle unfractionated malate dehydrogenase (MDH, EC 1.1.1.37 and its isolated isoforms were analyzed to further study the possible sMDH-A* locus duplication evolved from a recent tandem duplication. Both A (A1 and A2 and B isoforms had similar optima pH (7.5-8.0. While Hoplias A isoform could not be characterized as thermostable, B could as thermolabile. A isoforms differed from B isoform in having higher Km values for oxaloacetate. The possibly duplicated A2 isoform showed higher substrate affinity than the A1. Hoplias duplicated A isoforms may influence the direction of carbon flow between glycolisis and gluconeogenesis.

  18. Characterisation of CDKL5 Transcript Isoforms in Human and Mouse.

    Directory of Open Access Journals (Sweden)

    Ralph D Hector

    Full Text Available Mutations in the X-linked Cyclin-Dependent Kinase-Like 5 gene (CDKL5 cause early onset infantile spasms and subsequent severe developmental delay in affected children. Deleterious mutations have been reported to occur throughout the CDKL5 coding region. Several studies point to a complex CDKL5 gene structure in terms of exon usage and transcript expression. Improvements in molecular diagnosis and more extensive research into the neurobiology of CDKL5 and pathophysiology of CDKL5 disorders necessitate an updated analysis of the gene. In this study, we have analysed human and mouse CDKL5 transcript patterns both bioinformatically and experimentally. We have characterised the predominant brain isoform of CDKL5, a 9.7 kb transcript comprised of 18 exons with a large 6.6 kb 3'-untranslated region (UTR, which we name hCDKL5_1. In addition we describe new exonic regions and a range of novel splice and UTR isoforms. This has enabled the description of an updated gene model in both species and a standardised nomenclature system for CDKL5 transcripts. Profiling revealed tissue- and brain development stage-specific differences in expression between transcript isoforms. These findings provide an essential backdrop for the diagnosis of CDKL5-related disorders, for investigations into the basic biology of this gene and its protein products, and for the rational design of gene-based and molecular therapies for these disorders.

  19. Characterisation of CDKL5 Transcript Isoforms in Human and Mouse.

    Science.gov (United States)

    Hector, Ralph D; Dando, Owen; Landsberger, Nicoletta; Kilstrup-Nielsen, Charlotte; Kind, Peter C; Bailey, Mark E S; Cobb, Stuart R

    2016-01-01

    Mutations in the X-linked Cyclin-Dependent Kinase-Like 5 gene (CDKL5) cause early onset infantile spasms and subsequent severe developmental delay in affected children. Deleterious mutations have been reported to occur throughout the CDKL5 coding region. Several studies point to a complex CDKL5 gene structure in terms of exon usage and transcript expression. Improvements in molecular diagnosis and more extensive research into the neurobiology of CDKL5 and pathophysiology of CDKL5 disorders necessitate an updated analysis of the gene. In this study, we have analysed human and mouse CDKL5 transcript patterns both bioinformatically and experimentally. We have characterised the predominant brain isoform of CDKL5, a 9.7 kb transcript comprised of 18 exons with a large 6.6 kb 3'-untranslated region (UTR), which we name hCDKL5_1. In addition we describe new exonic regions and a range of novel splice and UTR isoforms. This has enabled the description of an updated gene model in both species and a standardised nomenclature system for CDKL5 transcripts. Profiling revealed tissue- and brain development stage-specific differences in expression between transcript isoforms. These findings provide an essential backdrop for the diagnosis of CDKL5-related disorders, for investigations into the basic biology of this gene and its protein products, and for the rational design of gene-based and molecular therapies for these disorders.

  20. Differential expression of syndecan isoforms during mouse incisor amelogenesis.

    Science.gov (United States)

    Muto, Taro; Miyoshi, Keiko; Munesue, Seiichi; Nakada, Hiroshi; Okayama, Minoru; Matsuo, Takashi; Noma, Takafumi

    2007-08-01

    Syndecans are transmembranous heparan sulfate proteoglycans (HSPGs) with covalently attached glycosaminoglycan side-chains located on the cell surface. The mammalian syndecan family is composed of four types of syndecans (syndecan-1 to -4). Syndecans interact with the intracellular cytoskeleton through the cytoplasmic domains of their core proteins and membrane proteins, extracellular enzymes, growth factors, and matrix components, through their heparan-sulfate chains, to regulate developmental processes.Here, as a first step to assess the possible roles of syndecan proteins in amelogenesis, we examined the expression patterns of all syndecan isoforms in continuously growing mouse incisors, in which we can overview major differentiation stages of amelogenesis at a glance. Understanding the expression domain of each syndecan isoform during specific developmental stages seems useful for investigating their physiological roles in amelogenesis. Immunohistochemical analysis of syndecan core proteins in the lower incisors from postnatal day 1 mice revealed spatially and temporally specific expression patterns, with syndecan-1 expressed in undifferentiated epithelial and mesenchymal cells, and syndecan-2, -3, and -4 in more differentiated cells. These findings suggest that each syndecan isoform functions distinctly during the amelogenesis of the incisors of mice.

  1. Antagonistic functions of two stardust isoforms in Drosophila photoreceptor cells.

    Science.gov (United States)

    Bulgakova, Natalia A; Rentsch, Michaela; Knust, Elisabeth

    2010-11-15

    Membrane-associated guanylate kinases (MAGUKs) are scaffolding proteins that organize supramolecular protein complexes, thereby partitioning the plasma membrane into spatially and functionally distinct subdomains. Their modular organization is ideally suited to organize protein complexes with cell type- or stage-specific composition, or both. Often more than one MAGUK isoform is expressed by one gene in the same cell, yet very little is known about their individual in vivo functions. Here, we show that two isoforms of Drosophila stardust, Sdt-H (formerly called Sdt-B2) and Sdt-D, which differ in their N terminus, are expressed in adult photoreceptors. Both isoforms associate with Crumbs and PATJ, constituents of the conserved Crumbs-Stardust complex. However, they form distinct complexes, localized at the stalk, a restricted region of the apical plasma membrane. Strikingly, Sdt-H and Sdt-D have antagonistic functions. While Sdt-H overexpression increases stalk membrane length and prevents light-dependent retinal degeneration, Sdt-D overexpression reduces stalk length and enhances light-dependent retinal degeneration. These results suggest that a fine-tuned balance of different Crumbs complexes regulates photoreceptor homeostasis.

  2. 75 FR 61700 - Purified Carboxymethylcellulose From Finland, the Netherlands, and Sweden: Final Results of the...

    Science.gov (United States)

    2010-10-06

    ... International Trade Administration Purified Carboxymethylcellulose From Finland, the Netherlands, and Sweden... purified carboxymethylcellulose (CMC) from, inter alia, Finland, the Netherlands, and Sweden, pursuant to... (120-day) sunset reviews of the Finland, the Netherlands, and Sweden antidumping duty orders...

  3. 75 FR 39207 - Purified Carboxymethylcellulose From Finland: Extension of Time Limit for Preliminary Results of...

    Science.gov (United States)

    2010-07-08

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE International Trade Administration Purified Carboxymethylcellulose From Finland: Extension of Time Limit for... 30, 2010. See Purified Carboxymethylcellulose From Finland: Extension of Time Limit for...

  4. 77 FR 14733 - Purified Carboxymethylcellulose From Finland and the Netherlands: Extension of Time Limit for...

    Science.gov (United States)

    2012-03-13

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE International Trade Administration Purified Carboxymethylcellulose From Finland and the Netherlands: Extension..., inter alia, purified carboxymethylcellulose from Finland and the Netherlands covering the period July...

  5. A spinach O-acetylserine(thiollyase homologue, SoCSaseLP, suppresses cysteine biosynthesis catalysed by other enzyme isoforms

    Directory of Open Access Journals (Sweden)

    Miki Noda

    2016-06-01

    Full Text Available An enzyme, O-acetylserine(thiollyase (OASTL, also known as O-acetylserine sulfhydrylase or cysteine synthase (CSase, catalyses the incorporation of sulfide into O-acetylserine and produces cysteine. We previously identified a cDNA encoding an OASTL-like protein from Spinacia oleracea, (SoCSaseLP, but a recombinant SoCSaseLP produced in Escherichia coli did not show OASTL activity. The exon-intron structure of the SoCSaseLP gene shared conserved structures with other spinach OASTL genes. The SoCSaseLP and a Beta vulgaris homologue protein, KMT13462, comprise a unique clade in the phylogenetic tree of the OASTL family. Interestingly, when the SoCSaseLP gene was expressed in tobacco plants, total OASTL activity in tobacco leaves was reduced. This reduction in total OASTL activity was most likely caused by interference by SoCSaseLP with cytosolic OASTL. To investigate the possible interaction of SoCSaseLP with a spinach cytosolic OASTL isoform SoCSaseA, a pull-down assay was carried out. The recombinant glutathione S-transferase (GST-SoCSaseLP fusion protein was expressed in E. coli together with the histidine-tagged SoCSaseA protein, and the protein extract was subjected to glutathione affinity chromatography. The histidine-tagged SoCSaseA was co-purified with the GST-SoCSaseLP fusion protein, indicating the binding of SoCSaseLP to SoCSaseA. Consistent with this interaction, the OASTL activity of the co-purified SoCSaseA was reduced compared with the activity of SoCSaseA that was purified on its own. These results strongly suggest that SoCSaseLP negatively regulates the activity of other cytosolic OASTL family members by direct interaction.

  6. Primary structure and tissue-specific expression of blue crab (Callinectes sapidus) metallothionein isoforms.

    Science.gov (United States)

    Brouwer, M; Enghild, J; Hoexum-Brouwer, T; Thogersen, I; Truncali, A

    1995-01-01

    In aquatic animals, synthesis of the metal-binding protein metallothionein (MT) can be induced through exposure to elevated levels of metals in food or water. Whether the different routes of exposure lead to expression of different metallothionein isoforms in different tissues in unknown. In this study we examined the induction of metallothionein isoforms in the hepatopancreas and gills of the blue crab Callinectes sapidus. When blue crabs are exposed to cadmium in their diet, the metal accumulates in the hepatopancreas. Size-exclusion and anion-exchange chromatography show the presence of five low-molecular-mass cadmium-binding proteins. All of the observed cadmium-binding proteins belong to the class I MT family. They are designated as MT-Ia, MT-Ib, MT-Ic, MT-IIa and MT-IIb. All purified proteins run as single peaks upon rechromatography on anion-exchange HPLC, except for MT-Ic, which segregates into two peaks corresponding to MT-Ia and MT-Ic. The amino acid sequence of MT-Ia and MT-Ic is identical. MT-Ib differs from MT-Ia and MT-Ic only in having an extra N-terminal methionine. The 18 cysteine residues in MT-Ia and MT-IIa occur in identical positions; however, of the remaining 40 amino acids, 15 are found to be different. MT-IIb is identical with MT-IIa, except for an extra methionine residue at its N-terminal position. It appears therefore that, of the five observed CdMTs, only two are the products of distinct genes. CdMT-Ia and -IIa are posttranslationally modified forms of Ib and IIb, respectively, and CdMT-Ia and -Ic appear to be conformational isomers. Cadmium-induced expression of the two genes is tissue-specific. When crabs are exposed to cadmium in water, the metal accumulates in the gills, where it is bound to MT-II. MT-I is virtually absent. PMID:7487904

  7. Serine protease isoforms in Gloydius intermedius venom: Full sequences, molecular phylogeny and evolutionary implications.

    Science.gov (United States)

    Yang, Zhang-Min; Yu, Hui; Liu, Zhen-Zhen; Pei, Jian-Zhu; Yang, Yu-E; Yan, Su-Xian; Zhang, Cui; Zhao, Wen-Long; Wang, Zhe-Zhi; Wang, Ying-Ming; Tsai, Inn-Ho

    2017-07-05

    Nine distinct venom serine proteases (vSPs) of Gloydius intermedius were studied by transcriptomic, sub-proteomic and phylogenetic analyses. Their complete amino acid sequences were deduced after Expression Sequence Tag (EST) analyses followed by cDNA cloning and sequencing. These vSPs appear to be paralogs and contain the catalytic triads and 1-4 potential N-glycosylation sites. Their relative expression levels evaluated by qPCR were grossly consistent with their EST hit-numbers. The major vSPs were purified by HPLC and their N-terminal sequences matched well to the deduced sequences, while fragments of the minor vSPs were detected by LC-MS/MS identification. Specific amidolytic activities of the fractions from HPLC and anion exchange separation were assayed using four chromogenic substrates, respectively. Molecular phylogenetic tree based on the sequences of these vSPs and their orthologs revealed six major clusters, one of them covered four lineages of plasminogen activator like vSPs. N-glycosylation patterns and variations for the vSPs are discussed. The high sequence similarities between G. intermedius vSPs and their respective orthologs from American pitvipers suggest that most of the isoforms evolved before Asian pitvipers migrated to the New World. Our results also indicate that the neurotoxic venoms contain more kallikrein-like vSPs and hypotensive components than the hemorrhagic venoms. Full sequences and expression levels of nine paralogous serine proteases (designated as GiSPs) of Gloydius intermedius venom have been studied. A kallikrein-like enzyme is most abundant and four isoforms homologous to venom plasminogen-activators are also expressed in this venom. Taken together, the present and previous data demonstrate that the neurotoxic G. intermedius venoms contain more hypotensive vSPs relative to other hemorrhagic pitviper venoms and the pitviper vSPs are highly versatile and diverse. Their structure-function relationships remain to be explored and

  8. X-ray diffraction study of highly purified human ceruloplasmin

    Science.gov (United States)

    Samygina, V. R.; Sokolov, A. V.; Pulina, M. O.; Bartunik, H. D.; Vasil'Ev, V. B.

    2008-07-01

    The three-dimensional structure of ceruloplasmin (CP) with unoccupied labile metal-binding sites and the structure of CP containing Ni2+ in the labile sites were solved for the first time at 2.6 and 2.95 Å resolution, respectively. Crystallization was performed with the use of storage-stable CP, which was prepared in the presence of proteinase inhibitors and purified from (pre)proteinases. Ceruloplasmin with Ni2+ crystallized in the orthorhombic space group, which had been earlier unknown for CP. Ceruloplasmin with the unoccupied labile sites crystallized in the trigonal crystal form. The differences in intermolecular contacts observed in the trigonal and orthorhombic crystal structures of CP are considered. The conformational changes attendant upon Ni2+ binding are described. It was suggested that the labile sites are multifunctional and can both bind metal ions potentially toxic to organisms and be involved in electron transfer from substrates to the active site.

  9. A method for purifying butyric crude oil fractions

    Energy Technology Data Exchange (ETDEWEB)

    Saskovets, V.V.; Gayle, A.A.; Proskuryakov, V.A.; Semenov, L.V.; Zakharov, A.P.

    1983-01-01

    In a method for purification of butyric fractions of oil through extraction by a selective solvent, in order to increase the output and to improve the quality of the purified oil, 2,5-dimethyl-1,3,4-oxadiazole of the cited formula, or its mixture with 80 to 90 percent furfural is used as the selective solvent. The solvent is produced through a reaction between hydrazine and an acetic anhydride. The solvent is a colorless liquid with a weak characteristic smell, and is easily dissolved in water with a boiling point of 178 degrees and density at 4-20/sup 0/ of 1.0963. The solvent is thermally stable: after boiling at 220 degrees, its viscosity is essentially the same.

  10. Using ion exchange chromatography to purify a recombinantly expressed protein.

    Science.gov (United States)

    Duong-Ly, Krisna C; Gabelli, Sandra B

    2014-01-01

    Ion exchange chromatography (IEX) separates molecules by their surface charge, a property that can vary vastly between different proteins. There are two types of IEX, cation exhange and anion exchange chromatography. The protocol that follows was designed by the authors for anion exchange chromatography of a recombinantly expressed protein having a pI of 4.9 and containing two cysteine residues and one tryptophan residue, using an FPLC system. Prior to anion exchange, the protein had been salted out using ammonium sulfate precipitation and partially purified via hydrophobic interaction chromatography (see Salting out of proteins using ammonium sulfate precipitation and Use and Application of Hydrophobic Interaction Chromatography for Protein Purification). Slight modifications to this protocol may be made to accommodate both the protein of interest and the availability of equipment.

  11. Robust sparse image reconstruction of radio interferometric observations with PURIFY

    CERN Document Server

    Pratley, Luke; d'Avezac, Mayeul; Carrillo, Rafael E; Onose, Alexandru; Wiaux, Yves

    2016-01-01

    Next-generation radio interferometers, such as the Square Kilometre Array (SKA), will revolutionise our understanding of the universe through their unprecedented sensitivity and resolution. However, to realise these goals significant challenges in image and data processing need to be overcome. The standard methods in radio interferometry for reconstructing images, such as CLEAN and its variants, have served the community well over the last few decades and have survived largely because they are pragmatic. However, they produce reconstructed interferometric images that are limited in quality and they are not scalable for big data. In this work we apply and evaluate alternative interferometric reconstruction methods that make use of state-of-the-art sparse image reconstruction algorithms motivated by compressive sensing, which have been implemented in the PURIFY software package. In particular, we implement and apply the proximal alternating direction method of multipliers (P-ADMM) algorithm presented in a recen...

  12. Techniques applicable for purifying Chironex fleckeri (box-jellyfish) venom.

    Science.gov (United States)

    Othman, I; Burnett, J W

    1990-01-01

    A survey of several techniques to isolate a purified lethal factor from the tentacles of Chironex fleckeri was completed. Heterologous band patterns were obtained from specific eluates after gel filtration, ion exchange, immunoaffinity and hydrophobic chromatography. SDS-PAGE revealed a dense band at 24,000 mol. wt in many of these fractions. Isoelectric focusing of the crude venom resulted in considerable loss of activity but indicated significant purification in the fractions having a pI of 5.2-6.8. These fractions were also immunologically active against sera from a convalescing post-evenomation patient. The primary difficulties encountered in jellyfish venom purification are the lack of stability and the tendency of the active toxins to adhere to each other and to various support matrices.

  13. Are whole extracts and purified glucosinolates from cruciferous vegetables antioxidants?

    Science.gov (United States)

    Plumb, G W; Lambert, N; Chambers, S J; Wanigatunga, S; Heaney, R K; Plumb, J A; Aruoma, O I; Halliwell, B; Miller, N J; Williamson, G

    1996-07-01

    Fruits and vegetables contain several classes of compounds that can potentially contribute to antioxidant activity, including vitamins, simple and complex phenolics, sulphur-containing compounds and glucosinolates. The glucosinolates are found in high concentration in many cruciferous vegetables, and it is well established that their breakdown products induce endogenous antioxidant defences such as quinone reductase and glutathione S-transferase in cells and in vivo. Despite the anticarcinogenic effect of these compounds in animal models, the direct antioxidant properties of this class of compounds have not been systematically studied. We therefore examined the free radical-scavenging properties of representative extracts and of purified glucosinolates from cruciferous vegetables, by measuring their effect on ascorbate- or NADPH/iron-induced peroxidation of human liver microsomes, ascorbate/iron-induced peroxidation on phospholipid liposomes, iron chelation and hydroxyl radical scavenging using the deoxyribose assay, total antioxidant potential using ABTS (2,2'-azinobis(3-ethyl-benzothiazoline-6-sulphonate)) and the bleomycin assay. Most of the extracts from cruciferous vegetables exhibited some antioxidant properties, although extracts from cooked Brussels sprouts increased the rate of microsomal lipid peroxidation. The effects in these assays were dependent upon processing and species of crucifer, and the glucosinolate content appeared to play a minor role in these effects, since purified glucosinolates exhibited only weak antioxidant properties. The total antioxidant activities of extracts from cooked and autolysed Brussels sprouts were identical within experimental error. This is probably due to the content of phenolics which is unaltered by autolysis, despite the differences between these samples in other assays especially NADPH-iron-induced lipid peroxidation of human liver microsomes. The results demonstrate that glucosinolates are unlikely to account for

  14. Neutral semi-purified glycerin in starting pigs feeding

    Directory of Open Access Journals (Sweden)

    Adriana Gomez Gallego

    2014-10-01

    Full Text Available Glycerin is a major co-product resulting from biodiesel production, and it has been proposed as a highenergy source for use in swine diets. However, it is necessary to determine the nutritional value of neutral semi-purified glycerin (NSPG. In this study two experiments were carried out to determine the nutritional value, evaluated the performance and economic feasibility of starting piglets fed on neutral semi-purified glycerin. A digestibility trial (Experiment I was conducted using 30 crossbred barrows with an initial average body weight of 42.91±1.58 kg. The glycerin levels used in the digestibility assay were 4, 8, 12 and 16% of the basal diet (corn + soybean meal based. The digestible energy (DE and metabolizable (ME energy values of glycerin were estimated by regression of DE and ME (kcal/kg intake associated with glycerin vs. glycerin intake (kg. The values (as-fed-basis of DE and ME (kcal/ kg obtained were 3,298 and 2,531, respectively. In Experiment II, 100 piglets (50 gilts and 50 barrows with BW = 15.14±0.06 to 30.28±0.65 were allotted in a randomized design using four inclusion levels (3.5, 7.0, 10.5 and 14% of NSPG. There were ten replicates with two piglets per experimental unit. Additionally, a control diet containing no glycerin (0% was formulated. The results show it is feasible to use up to 14% NSPG in piglet feed without impairing performance and plasma chemistry.

  15. Magnetism for understanding catalyst analysis of purified carbon nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Bellouard, Christine; Mercier, Guillaume; Cahen, Sébastien; Ghanbaja, Jaafar; Medjahdi, Ghouti [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France); Gleize, Jérôme [Laboratoire de Chimie Physique-Approche Multi-échelle de Milieux Complexes-Université de Lorraine, 1 Bd Arago, 57078 Metz (France); Lamura, Gianrico [CNR-SPIN – Dipartimento di Fisica, via Dodecaneso 33, 16146 Genova (Italy); Hérold, Claire [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France); Vigolo, Brigitte, E-mail: Brigitte.Vigolo@univ-lorraine.fr [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France)

    2016-08-01

    The precise quantification of catalyst residues in purified carbon nanotubes is often a major issue in view of any fundamental and/or applicative studies. More importantly, since the best CNTs are successfully grown with magnetic catalysts, their quantification becomes strictly necessary to better understand intrinsic properties of CNT. For these reasons, we have deeply analyzed the catalyst content remained in nickel–yttrium arc-discharge single walled carbon nanotubes purified by both a chlorine-gas phase and a standard acid-based treatment. The study focuses on Ni analysis which has been investigated by transmission electron microscopy, X-ray diffraction, thermogravimetry analysis, and magnetic measurements. In the case of the acid-based treatment, all quantifications result in a decrease of the nanocrystallized Ni by a factor of two. In the case of the halogen gas treatment, analysis and quantification of Ni content is less straightforward: a huge difference appears between X-ray diffraction and thermogravimetry results. Thanks to magnetic measurements, this disagreement is explained by the presence of Ni{sup 2+} ions, belonging to NiCl{sub 2} formed during the Cl-based purification process. In particular, NiCl{sub 2} compound appears under different magnetic/crystalline phases: paramagnetic or diamagnetic, or well intercalated in between carbon sheets with an ordered magnetic phase at low temperature. - Highlights: • Cl-gas treatment of Ni catalyst of carbon nanotubes leads to NiCl{sub 2} residue. • Magnetic measurements show the transformation of Ni{sup 0} in Ni{sup 2+}through a purification process. • High temperature Cl treatment removes 75% of metallic impurities. • Cl-purification yields to an amount of metal of 1.5% in arc-discharge CNT samples.

  16. 42 CFR 84.254 - Powered air-purifying respirators; requirements and tests.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Powered air-purifying respirators; requirements and... DEVICES Special Use Respirators § 84.254 Powered air-purifying respirators; requirements and tests. (a... air-purifying respirators prescribed in subpart L of this part are applicable to vinyl...

  17. 42 CFR 84.179 - Non-powered air-purifying particulate respirators; filter identification.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Non-powered air-purifying particulate respirators... RESPIRATORY PROTECTIVE DEVICES Non-Powered Air-Purifying Particulate Respirators § 84.179 Non-powered air-purifying particulate respirators; filter identification. (a) The respirator manufacturer, as part of...

  18. Measurement of Ozone Emission and Particle Removal Rates from Portable Air Purifiers

    Science.gov (United States)

    Mang, Stephen A.; Walser, Maggie L.; Nizkorodov, Sergey A.; Laux, John M.

    2009-01-01

    Portable air purifiers are popular consumer items, especially in areas with poor air quality. Unfortunately, most users of these air purifiers have minimal understanding of the factors affecting their efficiency in typical indoor settings. Emission of the air pollutant ozone (O[subscript 3]) by certain air purifiers is of particular concern. In an…

  19. Human placenta expresses both peripheral and neuronal isoform of tryptophan hydroxylase.

    Science.gov (United States)

    Laurent, Laetitia; Deroy, Kathy; St-Pierre, Joey; Côté, Francine; Sanderson, J Thomas; Vaillancourt, Cathy

    2017-09-01

    The role of placental serotonin has been an active topic of research notably because of its crucial role in brain development. However, which cell types synthesize serotonin in human placenta remains unknown. Moreover, it is not known if the two tryptophan hydroxylase isoforms (TPH1 and TPH2), the rate-limiting enzymes in serotonin biosynthesis, are expressed in placenta. Human placentas were obtained in first trimester or at term, and trophoblast cells were isolated and purified using a magnetic cell sorter and placed in primary culture. The tissue sublocalization of each TPH was determined by immunohistochemistry. TPH expression in primary villous trophoblasts was determined by PCR and immunoblotting, and serotonin secretion by LC-MS/MS. Villous cytotrophoblasts, syncytiotrophoblast, fetal capillaries, extravillous cytotrophoblasts, and decidual cells co-expressed TPH1 and TPH2. Moreover, mRNA and protein levels of both TPHs were detected in human primary trophoblast as well as in mouse placental tissues. Finally, human trophoblast cells were shown to produce serotonin de novo. This study demonstrates that both TPH1 and TPH2 are expressed in human and mouse placenta throughout pregnancy and helps to better understand the placental serotonin system, which is crucial for healthy pregnancy and fetal development. It is therefore important to further understand regulation of the placental serotonin system and how its disruption during pregnancy may impact the developing fetus and subsequent child programming. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  20. PSA Isoforms' Velocities for Early Diagnosis of Prostate Cancer.

    Science.gov (United States)

    Heidegger, Isabel; Klocker, Helmut; Pichler, Renate; Horninger, Wolfgang; Bektic, Jasmin

    2015-06-01

    Free prostate-specific antigen (fPSA) and its molecular isoforms are suggested for enhancement of PSA testing in prostate cancer (PCa). In the present study we evaluated whether PSA isoforms' velocities might serve as a tool to improve early PCa diagnosis. Our study population included 381 men who had undergone at least one ultrasound-guided prostate biopsy whose pathologic examination yielded PCa or showed no evidence of prostatic malignancy. Serial PSA, fPSA, and proPSA measurements were performed on serum samples covering 7 years prior to biopsy using Beckmann Coulter Access immunoassays. Afterwards, velocities of PSA (PSAV), fPSA% (fPSA%V), proPSA% (proPSA%V) and the ratio proPSA/PSA/V were calculated and their ability to discriminate cancer from benign disease was evaluated. Among 381 men included in the study, 202 (53%) were diagnosed with PCa and underwent radical prostatectomy at our Department. PSAV, fPSA%V, proPSA%V as well as proPSA/PSA/V were able to differentiate significantly between PCa and non-cancerous prostate. The highest discriminatory power between cancer and benign disease has been observed two and one year prior to diagnosis with all measured parameters. Among all measured parameters, fPSA%V showed the best cancer specificity of 45.3% with 90% of sensitivity. In summary, our results highlight the value of PSA isoforms' velocity for early detection of PCa. Especially fPSA%V should be used in the clinical setting to increase cancer detection specificity.

  1. Identification of an alternative splicing isoform of chicken Lmbr1.

    Science.gov (United States)

    Huang, Yanqun; Chen, Wen; Li, Ning; Deng, Xuemei; Kang, Xiangtao; Liu, Xiaojun

    2011-10-01

    Lmbr1 is the key candidate gene for limb development. Until now, at least five and four alternative splicing isoforms of Lmbr1 gene have been found in human and mouse, respectively. However, only two alternative splicing isoforms of this homologous gene have been reported in chicken. In the present study, one novel chicken Lmbr1 transcript variant (designated Lmbr1-1) was identified by 5' RACE and RT-PCR. Chicken Lmbr1-1 possesses one novel transcription start site different from Lmbr1-N, and was predicted to encode one 192 amino acid protein with length variation in comparison with chicken LMBR1-N protein, which was produced by 5' spliced site variation of chicken Lmbr1-N exon 10. Comparing with Lmbr1-N transcript, chicken Lmbr1-1 exhibited restricted tissue distribution of the expression. Comparative sequence analysis revealed a highly conservative intron element between chicken and mammalians from the intron 9 of chicken Lmbr1-N, indicating their possible importance as intronic elements in the regulation of alternative splicing of Lmbr1 in vertebrates. By direct PCR sequencing the exon 10 and its flanking sequences in chicken Lmbr1-N, four variation sites/haplotypes were identified from six chicken breeds. One 797A/G nonsynonymous mutation (266Arg/Gln) locating in exon 10 of chicken Lmbr1-N was predicted to affect the exon splice enhancer motif for serine/arginine-rich protein recognition. These data demonstrated that chicken Lmbr1 was alternatively spliced to generate multiple splice forms, as was the case in mammals and each of the alternative splicing isoforms might function differentially.

  2. The role of wild type RAS isoforms in cancer.

    Science.gov (United States)

    Zhou, Bingying; Der, Channing J; Cox, Adrienne D

    2016-10-01

    Mutationally activated RAS proteins are critical oncogenic drivers in nearly 30% of all human cancers. As with mutant RAS, the role of wild type RAS proteins in oncogenesis, tumour maintenance and metastasis is context-dependent. Complexity is introduced by the existence of multiple RAS genes (HRAS, KRAS, NRAS) and protein "isoforms" (KRAS4A, KRAS4B), by the ever more complicated network of RAS signaling, and by the increasing identification of numerous genetic aberrations in cancers that do and do not harbour mutant RAS. Numerous mouse model carcinogenesis studies and examination of patient tumours reveal that, in RAS-mutant cancers, wild type RAS proteins are likely to serve as tumour suppressors when the mutant RAS is of the same isoform. This evidence is particularly robust in KRAS mutant cancers, which often display suppression or loss of wild type KRAS, but is not as strong for NRAS. In contrast, although not yet fully elucidated, the preponderance of evidence indicates that wild type RAS proteins play a tumour promoting role when the mutant RAS is of a different isoform. In non-RAS mutant cancers, wild type RAS is recognized as a mediator of oncogenic signaling due to chronic activation of upstream receptor tyrosine kinases that feed through RAS. Additionally, in the absence of mutant RAS, activation of wild type RAS may drive cancer upon the loss of negative RAS regulators such as NF1 GAP or SPRY proteins. Here we explore the current state of knowledge with respect to the roles of wild type RAS proteins in human cancers.

  3. Modulation of Progesterone Receptor Isoform Expression in Pregnant Human Myometrium

    Directory of Open Access Journals (Sweden)

    Marina Ilicic

    2017-01-01

    Full Text Available Background. Regulation of myometrial progesterone receptor (PR expression is an unresolved issue central to understanding the mechanism of functional progesterone withdrawal and initiation of labor in women. Objectives. To determine whether pregnant human myometrium undergoes culture-induced changes in PR isoform expression ex situ and, further, to determine if conditions approaching the in vivo environment stabilise PR isoform expression in culture. Methods. Term nonlaboring human myometrial tissues were cultured under specific conditions: serum supplementation, steroids, stretch, cAMP, PMA, PGF2α, NF-κB inhibitors, or TSA. Following 48 h culture, PR-T, PR-A, and PR-B mRNA levels were determined using qRT-PCR. PR-A/PR-B ratios were calculated. Results. PR-T and PR-A expression and the PR-A/PR-B ratio significantly increased in culture. Steroids prevented the culture-induced increase in PR-T and PR-A expression. Stretch blocked the effects of steroids on PR-T and PR-A expression. PMA further increased the PR-A/PR-B ratio, while TSA blocked culture-induced increases of PR-A expression and the PR-A/PR-B ratio. Conclusion. Human myometrial tissue in culture undergoes changes in PR gene expression consistent with transition toward a laboring phenotype. TSA maintained the nonlaboring PR isoform expression pattern. This suggests that preserving histone and/or nonhistone protein acetylation is critical for maintaining the progesterone dependent quiescent phenotype of human myometrium in culture.

  4. Modulation of Progesterone Receptor Isoform Expression in Pregnant Human Myometrium

    Science.gov (United States)

    2017-01-01

    Background. Regulation of myometrial progesterone receptor (PR) expression is an unresolved issue central to understanding the mechanism of functional progesterone withdrawal and initiation of labor in women. Objectives. To determine whether pregnant human myometrium undergoes culture-induced changes in PR isoform expression ex situ and, further, to determine if conditions approaching the in vivo environment stabilise PR isoform expression in culture. Methods. Term nonlaboring human myometrial tissues were cultured under specific conditions: serum supplementation, steroids, stretch, cAMP, PMA, PGF2α, NF-κB inhibitors, or TSA. Following 48 h culture, PR-T, PR-A, and PR-B mRNA levels were determined using qRT-PCR. PR-A/PR-B ratios were calculated. Results. PR-T and PR-A expression and the PR-A/PR-B ratio significantly increased in culture. Steroids prevented the culture-induced increase in PR-T and PR-A expression. Stretch blocked the effects of steroids on PR-T and PR-A expression. PMA further increased the PR-A/PR-B ratio, while TSA blocked culture-induced increases of PR-A expression and the PR-A/PR-B ratio. Conclusion. Human myometrial tissue in culture undergoes changes in PR gene expression consistent with transition toward a laboring phenotype. TSA maintained the nonlaboring PR isoform expression pattern. This suggests that preserving histone and/or nonhistone protein acetylation is critical for maintaining the progesterone dependent quiescent phenotype of human myometrium in culture. PMID:28540297

  5. PKC isoforms interact with and phosphorylate DNMT1

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    Pradhan Sriharsa

    2011-05-01

    Full Text Available Abstract Background DNA methyltransferase 1 (DNMT1 has been shown to be phosphorylated on multiple serine and threonine residues, based on cell type and physiological conditions. Although recent studies have suggested that protein kinase C (PKC may be involved, the individual contribution of PKC isoforms in their ability to phosphorylate DNMT1 remains unknown. The PKC family consists of at least 12 isoforms that possess distinct differences in structure, substrate requirement, expression and localization. Results Here we show that PKCα, βI, βII, δ, γ, η, ζ and μ preferentially phosphorylate the N-terminal domain of human DNMT1. No such phosphorylation of DNMT1 was observed with PKCε. Using PKCζ as a prototype model, we also found that PKC physically interacts with and phosphorylates DNMT1. In vitro phosphorylation assays conducted with recombinant fragments of DNMT1 showed that PKCζ preferentially phosphorylated the N-terminal region of DNMT1. The interaction of PKCζ with DNMT1 was confirmed by GST pull-down and co-immunoprecipitation experiments. Co-localization experiments by fluorescent microscopy further showed that endogenous PKCζ and DNMT1 were present in the same molecular complex. Endogenous PKCζ activity was also detected when DNMT1 was immunoprecipitated from HEK-293 cells. Overexpression of both PKCζ and DNMT1 in HEK-293 cells, but not of either alone, reduced the methylation status of genes distributed across the genome. Moreover, in vitro phosphorylation of DNMT1 by PKCζ reduced its methytransferase activity. Conclusions Our results indicate that phosphorylation of human DNMT1 by PKC is isoform-specific and provides the first evidence of cooperation between PKCζ and DNMT1 in the control of the DNA methylation patterns of the genome.

  6. Permeation of Calcium through Purified Connexin 26 Hemichannels*

    Science.gov (United States)

    Fiori, Mariana C.; Figueroa, Vania; Zoghbi, Maria E.; Saéz, Juan C.; Reuss, Luis; Altenberg, Guillermo A.

    2012-01-01

    Gap junction channels communicate the cytoplasms of two cells and are formed by head to head association of two hemichannels, one from each of the cells. Gap junction channels and hemichannels are permeable to ions and hydrophilic molecules of up to Mr 1,000, including second messengers and metabolites. Intercellular Ca2+ signaling can occur by movement of a number of second messengers, including Ca2+, through gap junction channels, or by a paracrine pathway that involves activation of purinergic receptors in neighboring cells following ATP release through hemichannels. Understanding Ca2+ permeation through Cx26 hemichannels is important to assess the role of gap junction channels and hemichannels in health and disease. In this context, it is possible that increased Ca2+ influx through hemichannels under ischemic conditions contributes to cell damage. Previous studies suggest Ca2+ permeation through hemichannels, based on indirect arguments. Here, we demonstrate for the first time hemichannel permeability to Ca2+ by measuring Ca2+ transport through purified Cx26 hemichannels reconstituted in liposomes. We trapped the low affinity Ca2+-sensitive fluorescent probe Fluo-5N into the liposomes and followed the increases in intraliposomal [Ca2+] in response to an imposed [Ca2+] gradient. We show that Ca2+ does move through Cx26 hemichannels and that the permeability of the hemichannels to Ca2+ is high, similar to that for Na+. We suggest that hemichannels can be a significant pathway for Ca2+ influx into cells under conditions such as ischemia. PMID:23048025

  7. [Activity of purified diosmin in the treatment of hemorrhoids].

    Science.gov (United States)

    Diana, G; Catanzaro, M; Ferrara, A; Ferrari, P

    2000-01-01

    Several theories on the etio-pathogenesis and physio-pathology of hemorrhoids have been up to now proposed. From the fisio-pathological viewpoint, particular importance is retained by the vascular factor, which in its turn is influenced by mechanical and sphinceric factors, that impair the venous back-flow. In the evidence of an hemorrhoidal crisis, characterized by local oedema, pain and bleeding, the use of bioflavonoid drugs is deemed to be the first choice. We investigated the use of purified diosmin, given at a dose of two 450 mg tablets bid for the first 7 days, then at 1 tablet bid for up to 2 months, in a group of 66 patients suffering from primitive hemorrhoids of grade 1-4. Our results confirmed diosmin efficacy in decreasing both pain and bleeding: reduction rates of 79% and 67%, respectively, were reached in the first treatment week. In the second week, figures were 98% and 86%, respectively. Diosmin tolerability was excellent: this characteristic makes the drug very easy to handle by the general practitioner and also useful to the proctologist in the preparation of patient to further treatments.

  8. Purified and synthetic Alzheimer's amyloid beta (Aβ) prions.

    Science.gov (United States)

    Stöhr, Jan; Watts, Joel C; Mensinger, Zachary L; Oehler, Abby; Grillo, Sunny K; DeArmond, Stephen J; Prusiner, Stanley B; Giles, Kurt

    2012-07-03

    The aggregation and deposition of amyloid-β (Aβ) peptides are believed to be central events in the pathogenesis of Alzheimer's disease (AD). Inoculation of brain homogenates containing Aβ aggregates into susceptible transgenic mice accelerated Aβ deposition, suggesting that Aβ aggregates are capable of self-propagation and hence might be prions. Recently, we demonstrated that Aβ deposition can be monitored in live mice using bioluminescence imaging (BLI). Here, we use BLI to probe the ability of Aβ aggregates to self-propagate following inoculation into bigenic mice. We report compelling evidence that Aβ aggregates are prions by demonstrating widespread cerebral β-amyloidosis induced by inoculation of either purified Aβ aggregates derived from brain or aggregates composed of synthetic Aβ. Although synthetic Aβ aggregates were sufficient to induce Aβ deposition in vivo, they exhibited lower specific biological activity compared with brain-derived Aβ aggregates. Our results create an experimental paradigm that should lead to identification of self-propagating Aβ conformations, which could represent novel targets for interrupting the spread of Aβ deposition in AD patients.

  9. Common Wet Chemical Agents for Purifying Multiwalled Carbon Nanotubes

    Directory of Open Access Journals (Sweden)

    Rasel Das

    2014-01-01

    Full Text Available Purification and functionalization of multiwalled carbon nanotubes (MWCNTs are challenging but vital for their effective applications in various fields including water purification technologies, optoelectronics, biosensors, fuel cells, and electrode arrays. The currently available purification techniques, often complicated and time consuming, yielded shortened and curled MWCNTs that are not suitable for applications in certain fields such as membrane technologies, hybrid catalysis, optoelectronics, and sensor developments. Here we described the H2O2 synergy on the actions of HCl and KOH in purifying and functionalizing pristine MWCNTs. The method (HCl/H2O2 showed 100% purification yield as compared to HCl and KOH/H2O2 with purification yields 93.46 and 3.92%, respectively. We probed the findings using transmission electron microscope, energy dispersive X-ray spectroscope, attenuated total reflectance infrared spectroscope, Raman spectroscope, thermal gravimetric analysis, and X-ray powder diffraction. The study is a new avenue for simple, rapid, low cost, and scalable purification of pristine MWCNTs for application in versatile fields.

  10. Widespread purifying selection on RNA structure in mammals.

    Science.gov (United States)

    Smith, Martin A; Gesell, Tanja; Stadler, Peter F; Mattick, John S

    2013-09-01

    Evolutionarily conserved RNA secondary structures are a robust indicator of purifying selection and, consequently, molecular function. Evaluating their genome-wide occurrence through comparative genomics has consistently been plagued by high false-positive rates and divergent predictions. We present a novel benchmarking pipeline aimed at calibrating the precision of genome-wide scans for consensus RNA structure prediction. The benchmarking data obtained from two refined structure prediction algorithms, RNAz and SISSIz, were then analyzed to fine-tune the parameters of an optimized workflow for genomic sliding window screens. When applied to consistency-based multiple genome alignments of 35 mammals, our approach confidently identifies >4 million evolutionarily constrained RNA structures using a conservative sensitivity threshold that entails historically low false discovery rates for such analyses (5-22%). These predictions comprise 13.6% of the human genome, 88% of which fall outside any known sequence-constrained element, suggesting that a large proportion of the mammalian genome is functional. As an example, our findings identify both known and novel conserved RNA structure motifs in the long noncoding RNA MALAT1. This study provides an extensive set of functional transcriptomic annotations that will assist researchers in uncovering the precise mechanisms underlying the developmental ontologies of higher eukaryotes.

  11. PURIFIED WASTE FCC CATALYST AS A CEMENT REPLACEMENT MATERIAL

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    Danute Vaiciukyniene

    2015-06-01

    Full Text Available Zeolites are commonly used in the fluid catalytic cracking process. Zeolite polluted with oil products and became waste after some time used. The quantity of this waste inevitably rises by expanding rapidly oil industry. The composition of these catalysts depends on the manufacturer and on the process that is going to be used. The main factors retarding hydration process of cement systems and modifying them strength are organic compounds impurities in the waste FCC catalyst. The present paper shows the results of using purified waste FCC catalyst (pFCC from Lithuania oil refinery, as Portland cement replacement material. For this purpose, the purification of waste FCC catalyst (FCC samples was treated with hydrogen peroxide. Hydrogen peroxide (H2O2 is one of the most powerful oxidizers known. By acting of waste with H2O2 it can eliminate the aforementioned waste deficiency, and the obtained product becomes one of the most promising ingredients, in new advanced building materials. Hardened cement paste samples with FCC or pFCC were formed. It was observed that the pFCC blended cements developed higher strength, after 28 days, compared to the samples with FCC or reference samples. Typical content of Portland cement substituting does not exceed 30 % of mass of Portland cement in samples. Reducing the consumption of Portland cement with utilizing waste materials is preferred for reasons of environmental protection.

  12. Aflatoxin detoxification by manganese peroxidase purified from Pleurotus ostreatus

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    Ramy Sayed Yehia

    2014-01-01

    Full Text Available Manganese peroxidase (MnP was produced from white rot edible mushroom Pleurotus ostreatus on the culture filtrate. The enzyme was purified to homogeneity using (NH42SO4 precipitation, DEAE-Sepharose and Sephadex G-100 column chromatography. The final enzyme activity achieved 81UmL-1, specific activity 78 U mg-1 with purification fold of 130 and recovery 1.2% of the crude enzyme. SDS-PAGE indicated that the pure enzyme have a molecular mass of approximately 42 kDa. The optimum pH was between 4-5 and the optimum temperature was 25 ºC. The pure MnP activity was enhanced by Mn2+,Cu2+,Ca2+ and K+ and inhibited by Hg+2 and Cd+2.H2O2 at 5 mM enhanced MnP activity while at 10 mM inhibited it significantly. The MnP-cDNA encoding gene was sequenced and determined (GenBank accession no. AB698450.1. The MnP-cDNA was found to consist of 497 bp in an Open Reading Frame (ORF encoding 165 amino acids. MnP from P. ostreatus could detoxify aflatoxin B1 (AFB1 depending on enzyme concentration and incubation period. The highest detoxification power (90% was observed after 48 h incubation at 1.5 U mL-1 enzyme activities.

  13. Aflatoxin detoxification by manganese peroxidase purified from Pleurotus ostreatus.

    Science.gov (United States)

    Yehia, Ramy Sayed

    2014-01-01

    Manganese peroxidase (MnP) was produced from white rot edible mushroom Pleurotus ostreatus on the culture filtrate. The enzyme was purified to homogeneity using (NH4)2SO4 precipitation, DEAE-Sepharose and Sephadex G-100 column chromatography. The final enzyme activity achieved 81 U mL(-1), specific activity 78 U mg(-1) with purification fold of 130 and recovery 1.2% of the crude enzyme. SDS-PAGE indicated that the pure enzyme have a molecular mass of approximately 42 kDa. The optimum pH was between 4-5 and the optimum temperature was 25 °C. The pure MnP activity was enhanced by Mn(2+), Cu(2+), Ca(2+) and K(+) and inhibited by Hg(+2) and Cd(+2). H2O2 at 5 mM enhanced MnP activity while at 10 mM inhibited it significantly. The MnP-cDNA encoding gene was sequenced and determined (GenBank accession no. AB698450.1). The MnP-cDNA was found to consist of 497 bp in an Open Reading Frame (ORF) encoding 165 amino acids. MnP from P. ostreatus could detoxify aflatoxin B1 (AFB1) depending on enzyme concentration and incubation period. The highest detoxification power (90%) was observed after 48 h incubation at 1.5 U mL(-1) enzyme activities.

  14. Synaptosomal protein synthesis in P2 and Ficoll purified fractions.

    Science.gov (United States)

    Eyman, Maria; Cefaliello, Carolina; Bruno, Anna Paola; Bruno, Annapaola; Crispino, Marianna; Giuditta, Antonio

    2012-01-30

    Cytoplasmic protein synthesis of brain synaptosomes has generally been determined in the Ficoll purified fraction which contains fewer contaminating mitochondria, microsomes and myelin fragments than the parent P2 fraction. Using a highly selective assay of this activity we have compared the total translation activity and the specific activity of the proteins synthesized by either fraction in control rats and in rats trained for a two-way active avoidance task. In control rats the specific activity remained essentially the same in both fractions but in trained rats the value of the Ficoll fraction was markedly lower (38.5%) than in the P2 fraction. Furthermore, the total translation activity of the Ficoll fraction was 30% lower than in the P2 fraction in control rats and 62% lower in trained rats. These decrements indicate that a large proportion of active synaptosomes present in the P2 fraction is not recovered in the Ficoll fraction, notably in rats undergoing plastic brain changes. We conclude that cytoplasmic protein synthesis of brain synaptosomes is better preserved in the P2 fraction.

  15. Purifying effect of new flux on magnesium alloy

    Institute of Scientific and Technical Information of China (English)

    高洪涛; 吴国华; 丁文江; 朱燕萍

    2004-01-01

    A new flux which can remove both Fe and non-metallic inclusions in magnesium alloy was introduced.The Fe content of the magnesium alloy can be decreased greatly from 0. 062% to lower than 0. 005% (degree of AZ91D) after being purified by this new flux. The optimum addition of B2O3 in the flux is 0. 58 % by Gaussian Curve Fitting. Corrosion rate was measured after the specimen being immersed in 5 % NaCl solution for 3 d. The resuits show that the corrosion rate of the magnesium alloy after purification by the new flux is only 0.3 mg · cm-2 ·d-1. On the other hand, non-metallic inclusions in the magnesium alloy decrease with increasing addition of JDMJ in the new flux. Average volume fraction of the non-metallic inclusions in the magnesium alloy decreases from 1.52 % to 1.08%, which leads to improvement in the mechanical properties of the magnesium alloy by 30%. The mechanisms of Fe reduction and non-metallic inclusion-removing in magnesium melt by purification with the new flux were also revealed.

  16. Biological characterization of purified macrophage-derived neutrophil chemotactic factor

    Directory of Open Access Journals (Sweden)

    M. Dias-Baruffi

    1995-01-01

    Full Text Available We have recently described the purification of a 54 kDa acidic protein, identified as macrophage-derived neutrophil chemotactic factor (MNCF. This protein causes in vitro chemotaxis as well as in vivo neutrophil migration even in animals treated with dexamethasone. This in vivo chemotactic activity of MNCF in animals pretreated with dexamethasone is an uncommon characteristic which discriminates MNCF from known chemotactic cytokines. MNCF is released in the supernatant by macrophage monolayers stimulated with lipopolysaccharide (LPS. In the present study, we describe some biological characteristics of homogenous purified MNCF. When assayed in vitro, MNCF gave a bell-shaped dose–response curve. This in vitro activity was shown to be caused by haptotaxis. Unlike N-formyl-methionylleucyl- phenylalanine (FMLP or interleukin 8 (IL-8, the chemotactic activity of MNCF in vivo and in vitro, was inhibited by preincubation with D-galactose but not with D-mannose. In contrast with IL-8, MNCF did not bind to heparin and antiserum against IL-8 was ineffective in inhibiting its chemotactic activity. These data indicate that MNCF induces neutrophil migration through a carbohydrate recognition property, but by a mechanism different from that of the known chemokines. It is suggested that MNCF may be an important mediator in the recruitment of neutrophils via the formation of a substrate bound chemotactic gradient (haptotaxis in the inflamed tissues.

  17. Characterization of fructose 6 phosphate phosphoketolases purified from Bifidobacterium species.

    Science.gov (United States)

    Grill, J P; Crociani, J; Ballongue, J

    1995-07-01

    Fructose 6 phosphate phosphoketolases (F6PPKs) were purified from Bifidobacterium longum BB536, B. dentium ATCC 27534, B. globosum ATCC 25864, and Bifidobacterium animalis ATCC 25527. Concerning ions (Cu++, Zn++, Ca++, Mg++, Fe++, Co++, Mn++) and common enzyme inhibitors (fructose, ammonium sulfate, iodoacetate, and parachloromercuribenzoic acid), no difference appeared between the enzymes. Cu++, parachloromercuribenzoic acid (pCMB), and mercuric acetate induced high enzymatic inhibition. The study of pCMB demonstrated a noncompetitive inhibition. Additional results showed that the sulfhydryl group was not involved in catalytic reaction. Photooxidation experiments and determination of ionizable group pKas (5.16-7.17) suggested the presence of one or more histidines necessary for the catalytic reaction and explained the inhibition observed with pCMB. In light of the noncompetitive inhibition, this group was not directly involved in substrate binding. Determination of Km demonstrated that the affinities for fructose 6 phosphate in the case of animal and human origin strains were close. In addition, the same enzymatic efficiency (Kcat/Km) was obtained for each strain. The F6PPK activity was regulated by sodium pyrophosphate, ATP, and especially by ADP.

  18. Characterisation of the 1st SSI purified MBL standard

    DEFF Research Database (Denmark)

    Laursen, Inga; Højrup, Peter; Houen, Gunnar

    2008-01-01

    BACKGROUND: Mannan-binding lectin (MBL) is of importance in innate immunity. MBL-deficiency, the most common immune defect, is significant in several clinical contexts. The request for MBL diagnostic is increasing, hence we developed a high-purity MBL standard assigned with a traceable value. MET...... purified MBL standard has been produced, and assigned the value 192.6 microg MBL/ml, traceable to an accurate realisation of the unit.......BACKGROUND: Mannan-binding lectin (MBL) is of importance in innate immunity. MBL-deficiency, the most common immune defect, is significant in several clinical contexts. The request for MBL diagnostic is increasing, hence we developed a high-purity MBL standard assigned with a traceable value....... METHODS AND RESULTS: The standard material was produced from human plasma; and the protein concentration determined by amino acid analysis after a preceding desalting. The standard value was assessed by two series of sub-sample analyses from nine vials by the grand mean: 235.7 microg protein/ml (range 191...

  19. Analysis of cavitation effect for water purifier using electrolysis

    Science.gov (United States)

    Shin, Dong Ho; Ko, Han Seo; Lee, Seung Ho

    2015-11-01

    Water is a limited and vital resource, so it should not be wasted by pollution. A development of new water purification technology is urgent nowadays since the original and biological treatments are not sufficient. The microbubble-aided method was investigated for removal of algal in this study since it overcomes demerits of the existing purification technologies. Thus, the cavitation effect in a venturi-type tube using the electrolysis was analyzed. Ruthenium-coated titanium plates were used as electrodes. Optimum electrode interval and applied power were determined for the electrolysis. Then, the optimized electrodes were installed in the venturi-type tube for generating cavitation. The cavitation effect could be enhanced without any byproduct by the bubbly flow induced by the electrolysis. The optimum mass flow rate and current were determined for the cavitation with the electrolysis. Finally, the visualization techniques were used to count the cell number of algal and microbubbles for the confirmation of the performance. As a result, the energy saving and high efficient water purifier was fabricated in this study. This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Korean government (MEST) (No. 2013R1A2A2A01068653).

  20. The FU gene and its possible protein isoforms

    Directory of Open Access Journals (Sweden)

    Nöthen Markus M

    2004-07-01

    Full Text Available Abstract Background FU is the human homologue of the Drosophila gene fused whose product fused is a positive regulator of the transcription factor Cubitus interruptus (Ci. Thus, FU may act as a regulator of the human counterparts of Ci, the GLI transcription factors. Since Ci and GLI are targets of Hedgehog signaling in development and morphogenesis, it is expected that FU plays an important role in Sonic, Desert and/or Indian Hedgehog induced cellular signaling. Results The FU gene was identified on chromosome 2q35 at 217.56 Mb and its exon-intron organization determined. The human developmental disorder Syndactyly type 1 (SD1 maps to this region on chromosome 2 and the FU coding region was sequenced using genomic DNA from an affected individual in a linked family. While no FU mutations were found, three single nucleotide polymorphisms were identified. The expression pattern of FU was thoroughly investigated and all examined tissues express FU. It is also clear that different tissues express transcripts of different sizes and some tissues express more than one transcript. By means of nested PCR of specific regions in RT/PCR generated cDNA, it was possible to verify two alternative splicing events. This also suggests the existence of at least two additional protein isoforms besides the FU protein that has previously been described. This long FU and a much shorter isoform were compared for the ability to regulate GLI1 and GLI2. None of the FU isoforms showed any effects on GLI1 induced transcription but the long form can enhance GLI2 activity. Apparently FU did not have any effect on SUFU induced inhibition of GLI. Conclusions The FU gene and its genomic structure was identified. FU is a candidate gene for SD1, but we have not identified a pathogenic mutation in the FU coding region in a family with SD1. The sequence information and expression analyses show that transcripts of different sizes are expressed and subjected to alternative splicing

  1. Tubulin evolution in insects: gene duplication and subfunctionalization provide specialized isoforms in a functionally constrained gene family

    Directory of Open Access Journals (Sweden)

    Gadagkar Sudhindra R

    2010-04-01

    Full Text Available Abstract Background The completion of 19 insect genome sequencing projects spanning six insect orders provides the opportunity to investigate the evolution of important gene families, here tubulins. Tubulins are a family of eukaryotic structural genes that form microtubules, fundamental components of the cytoskeleton that mediate cell division, shape, motility, and intracellular trafficking. Previous in vivo studies in Drosophila find a stringent relationship between tubulin structure and function; small, biochemically similar changes in the major alpha 1 or testis-specific beta 2 tubulin protein render each unable to generate a motile spermtail axoneme. This has evolutionary implications, not a single non-synonymous substitution is found in beta 2 among 17 species of Drosophila and Hirtodrosophila flies spanning 60 Myr of evolution. This raises an important question, How do tubulins evolve while maintaining their function? To answer, we use molecular evolutionary analyses to characterize the evolution of insect tubulins. Results Sixty-six alpha tubulins and eighty-six beta tubulin gene copies were retrieved and subjected to molecular evolutionary analyses. Four ancient clades of alpha and beta tubulins are found in insects, a major isoform clade (alpha 1, beta 1 and three minor, tissue-specific clades (alpha 2-4, beta 2-4. Based on a Homarus americanus (lobster outgroup, these were generated through gene duplication events on major beta and alpha tubulin ancestors, followed by subfunctionalization in expression domain. Strong purifying selection acts on all tubulins, yet maximum pairwise amino acid distances between tubulin paralogs are large (0.464 substitutions/site beta tubulins, 0.707 alpha tubulins. Conversely orthologs, with the exception of reproductive tissue isoforms, show little sequence variation except in the last 15 carboxy terminus tail (CTT residues, which serve as sites for post-translational modifications (PTMs and interactions

  2. Soluble malate dehydrogenase of Geophagus brasiliensis (Cichlidae, Perciformes: isolated isoforms and kinetics properties

    Directory of Open Access Journals (Sweden)

    Maria Regina de Aquino-Silva

    2008-01-01

    Full Text Available Kinetic properties and thermal stabilities of Geophagus brasiliensis skeletal muscle unfractionated malate dehydrogenase (MDH, EC 1.1.1.37 and its isolated isoforms were analyzed to examine a possible sMDH-B* locus duplication in a fixation process influenced by genetic drift. Two optimal pHs were detected: 7.5 for AB1 unfractionated muscle phenotype and its B1 isoform, and 8.0 for AB1B2 unfractionated muscle phenotype, A and B2 isoforms. While G. brasiliensis A isoform could be characterized as thermostable, the duplicated B isoform cannot be assumed as thermolabile. Km values for isolated B2 isoforms were 1.6 times lower than for B1. A duplication event in progress best explains the electrophoretic six-band pattern detected in G. brasiliensis, which would be caused by genetic drift.

  3. Isoform-specific regulation of adenylyl cyclase: a potential target in future pharmacotherapy.

    Science.gov (United States)

    Iwatsubo, Kousaku; Tsunematsu, Takashi; Ishikawa, Yoshihiro

    2003-06-01

    Adenylyl cyclase (AC) is a target enzyme of multiple G-protein-coupled receptors (GPCRs). In the past decade, the cloning, structure and biochemical properties of nine AC isoforms were reported, and each isoform of AC shows distinct patterns of tissue distribution and biochemical/pharmacological properties. In addition to the conventional regulators of this enzyme, such as calmodulin (CaM) or PKC, novel regulators, for example, caveolin, have been identified. Most importantly, these regulators work on AC in an isoform dependent manner. Recent studies have demonstrated that certain classic AC inhibitors, i.e., P-site inhibitors, show an isoform-dependent inhibition of AC. The side chain modifications of forskolin, a diterpene extract from Coleus forskolii, markedly enhance its isoform selectivity. When taken together, these findings suggest that it is feasible to develop new pharmacotherapeutic agents that target AC isoforms to regulate various neurohormonal signals in a highly tissue-/organ-specific manner.

  4. Separation of arginase isoforms by capillary zone electrophoresis and isoelectric focusing in density gradient column.

    Science.gov (United States)

    Pedrosa, M M; Legaz, M E

    1995-04-01

    Four major arginase isoforms, I, II, III and IV, have been detected in Evernia prunastri thallus. They differ in terms of both physical and biochemical properties. The isoelectric point (pI) of these proteins has been determined by both isoelectric focusing in density gradient column and high-performance capillary electrophoresis (HPCE). Isoelectric focusing revealed charge microheterogeneity for isoforms II and IV whereas arginases I and II had the same pI value of 5.8. HPCE separation confirmed this charge microheterogeneity for isoform IV but not for isoform III, and provided evidence of microheterogeneity for isoforms I and II. The effect of various electrolyte buffers and running conditions on the HPCE separation of arginase isoform were investigated. Addition of 0.5 mM spermidine (SPD) to the running buffer reduced the electroosmotic flow (EOF) and permitted discriminating between the native proteins and protein fragments.

  5. CD44 variant isoforms in non-Hodgkin's lymphoma : a new independent prognostic factor

    OpenAIRE

    Stauder, R.; Eisterer, W.; Thaler, J; Günthert, U

    1995-01-01

    Isoforms of the transmembrane glycoprotein CD44, generated by alternative RNA splicing, have been correlated to tumor dissemination. For evaluation of the potential role of CD44 variant isoforms in non-Hodgkin's lymphoma (NHL), the presence of CD44 isoforms was analyzed in a large panel of reactive and neoplastic lymphoid tissues by immunohistochemical staining, as well as detection of CD44 variant RNAs by the reverse transcriptase-polymerase chain reaction. Whereas the CD44 standard or hemat...

  6. Identification of signals that facilitate isoform specific nucleolar localization of myosin IC

    Energy Technology Data Exchange (ETDEWEB)

    Schwab, Ryan S.; Ihnatovych, Ivanna; Yunus, Sharifah Z.S.A.; Domaradzki, Tera [Department of Physiology and Biophysics, University at Buffalo—State University of New York, Buffalo, NY (United States); Hofmann, Wilma A., E-mail: whofmann@buffalo.edu [Department of Physiology and Biophysics, University at Buffalo—State University of New York, Buffalo, NY (United States)

    2013-05-01

    Myosin IC is a single headed member of the myosin superfamily that localizes to the cytoplasm and the nucleus, where it is involved in transcription by RNA polymerases I and II, intranuclear transport, and nuclear export. In mammalian cells, three isoforms of myosin IC are expressed that differ only in the addition of short isoform-specific N-terminal peptides. Despite the high sequence homology, the isoforms show differences in cellular distribution, in localization to nuclear substructures, and in their interaction with nuclear proteins through yet unknown mechanisms. In this study, we used EGFP-fusion constructs that express truncated or mutated versions of myosin IC isoforms to detect regions that are involved in isoform-specific localization. We identified two nucleolar localization signals (NoLS). One NoLS is located in the myosin IC isoform B specific N-terminal peptide, the second NoLS is located upstream of the neck region within the head domain. We demonstrate that both NoLS are functional and necessary for nucleolar localization of specifically myosin IC isoform B. Our data provide a first mechanistic explanation for the observed functional differences between the myosin IC isoforms and are an important step toward our understanding of the underlying mechanisms that regulate the various and distinct functions of myosin IC isoforms. - Highlights: ► Two NoLS have been identified in the myosin IC isoform B sequence. ► Both NoLS are necessary for myosin IC isoform B specific nucleolar localization. ► First mechanistic explanation of functional differences between the isoforms.

  7. The Role of a Novel Myosin Isoform in Prostate Cancer Metastasis

    Science.gov (United States)

    2013-10-01

    specific N-terminal peptides. Despite the high sequence homology , the isoforms show differences in cellular distribution, in localization to nuclear...Interestingly, despite the sequence homology between the isoforms and the commonality of the NLS, the isoforms show differences in their nucleus to cytoplasm...Goeres J, Sixt KM, Bekes M, Zhang XD, Salvesen GS, Matunis MJ (2009). Protection from isopeptidase-mediated deconjugation regulates paralog -selective

  8. Squalamine, a novel cationic steroid, specifically inhibits the brush-border Na+/H+ exchanger isoform NHE3.

    Science.gov (United States)

    Akhter, S; Nath, S K; Tse, C M; Williams, J; Zasloff, M; Donowitz, M

    1999-01-01

    Squalamine, an endogenous molecule found in the liver and other tissues of Squalus acanthias, has antibiotic properties and causes changes in endothelial cell shape. The latter suggested that its potential targets might include transport proteins that control cell volume or cell shape. The effect of purified squalamine was examined on cloned Na+/H+ exchanger isoforms NHE1, NHE2, and NHE3 stably transfected in PS120 fibroblasts. Squalamine (1-h pretreatment) decreased the maximal velocity of rabbit NHE3 in a concentration-dependent manner (13, 47, and 57% inhibition with 3, 5, and 7 micrograms/ml, respectively) and also increased K'[H+]i. Squalamine did not affect rabbit NHE1 or NHE2 function. The inhibitory effect of squalamine was 1) time dependent, with no effect of immediate addition and maximum effect with 1 h of exposure, and 2) fully reversible. Squalamine pretreatment of the ileum for 60 min inhibited brush-border membrane vesicle Na+/H+ activity by 51%. Further investigation into the mechanism of squalamine's effects showed that squalamine required the COOH-terminal 76 amino acids of NHE3. Squalamine had no cytotoxic effect at the concentrations studied, as indicated by monitoring lactate dehydrogenase release. These results indicate that squalamine 1) is a specific inhibitor of the brush-border NHE isoform NHE3 and not NHE1 or NHE2, 2) acts in a nontoxic and fully reversible manner, and 3) has a delayed effect, indicating that it may influence brush-border Na+/H+ exchanger function indirectly, through an intracellular signaling pathway or by acting as an intracellular modulator.

  9. Cloning and Characterization of Two NAD Kinases from Arabidopsis. Identification of a Calmodulin Binding Isoform1[w

    Science.gov (United States)

    Turner, William L.; Waller, Jeffrey C.; Vanderbeld, Barb; Snedden, Wayne A.

    2004-01-01

    NAD kinase (NADK; ATP:NAD 2′-phosphotransferase, EC 2.7.1.23), an enzyme found in both prokaryotes and eukaryotes, generates the important pyridine nucleotide NADP from substrates ATP and NAD. The role of NADKs in plants is poorly understood, and cDNAs encoding plant NADKs have not previously been described to our knowledge. We have cloned two cDNAs from Arabidopsis predicted to encode NADK isoforms, designated NADK1 and NADK2, respectively. Expressed as recombinant proteins in bacteria, both NADK1 and NADK2 were catalytically active, thereby confirming their identity as NADKs. Transcripts for both isoforms were detected in all tissues examined and throughout development. Although the predicted catalytic regions for NADK1 and NADK2 show sequence similarity to NADKs from other organisms, NADK2 possesses a large N-terminal extension that appears to be unique to plants. Using recombinant glutathione-S-transferase fusion proteins and calmodulin (CaM)-affinity chromatography, we delineated a Ca2+-dependent CaM-binding domain to a 45-residue region within the N-terminal extension of NADK2. Although recombinant NADK2 was not responsive to CaM in vitro, immunoblot analysis suggests that native NADK2 is a CaM-binding protein. In Arabidopsis crude extracts, CaM-dependent NADK activity was much greater than CaM-independent activity throughout development, particularly in young seedlings. A native CaM-dependent NADK was partially purified from Arabidopsis seedlings (KmNAD = 0.20 mM, KmMg2+−ATP = 0.17 mM). The enzyme was fully activated by conserved CaM (S0.5 = 2.2 nm) in the presence of calcium but displayed differential responsiveness to eight CaM-like Arabidopsis proteins. Possible roles for NADKs in plants are discussed in light of our observations. PMID:15247403

  10. Nuclear progesterone receptor isoforms and their functions in the female reproductive tract.

    Science.gov (United States)

    Rekawiecki, R; Kowalik, M K; Kotwica, J

    2011-01-01

    Progesterone (P4), which is produced by the corpus luteum (CL), creates proper conditions for the embryo implantation, its development, and ensures proper conditions for the duration of pregnancy. Besides the non-genomic activity of P4 on target cells, its main physiological effect is caused through genomic action by the progesterone nuclear receptor (PGR). This nuclear progesterone receptor occurs in two specific isoforms, PGRA and PGRB. PGRA isoform acts as an inhibitor of transcriptional action of PGRB. The inactive receptor is connected with chaperone proteins and attachment of P4 causes disconnection of chaperones and unveiling of DNA binding domain (DBD). After receptor dimerization in the cells' nucleus and interaction with hormone response element (HRE), the receptor coactivators are connected and transcription is initiated. The ratio of these isoforms changes during the estrous cycle and reflects the different levels of P4 effect on the reproductive system. Both isoforms, PGRA and PGRB, also show a different response to the P4 receptor antagonist activity. Connection of the antagonist to PGRA can block PGRB, but acting through the PGRB isoform, P4 receptor antagonist may undergo conversion to a strongly receptor agonist. A third isoform, PGRC, has also been revealed. This isoform is the shortest and does not have transcriptional activity. Alternative splicing and insertion of additional exons may lead to the formation of different PGR isoforms. This paper summarizes the available data on the progesterone receptor isoforms and its regulatory action within the female reproductive system.

  11. Identification of signals that facilitate isoform specific nucleolar localization of myosin IC.

    Science.gov (United States)

    Schwab, Ryan S; Ihnatovych, Ivanna; Yunus, Sharifah Z S A; Domaradzki, Tera; Hofmann, Wilma A

    2013-05-01

    Myosin IC is a single headed member of the myosin superfamily that localizes to the cytoplasm and the nucleus, where it is involved in transcription by RNA polymerases I and II, intranuclear transport, and nuclear export. In mammalian cells, three isoforms of myosin IC are expressed that differ only in the addition of short isoform-specific N-terminal peptides. Despite the high sequence homology, the isoforms show differences in cellular distribution, in localization to nuclear substructures, and in their interaction with nuclear proteins through yet unknown mechanisms. In this study, we used EGFP-fusion constructs that express truncated or mutated versions of myosin IC isoforms to detect regions that are involved in isoform-specific localization. We identified two nucleolar localization signals (NoLS). One NoLS is located in the myosin IC isoform B specific N-terminal peptide, the second NoLS is located upstream of the neck region within the head domain. We demonstrate that both NoLS are functional and necessary for nucleolar localization of specifically myosin IC isoform B. Our data provide a first mechanistic explanation for the observed functional differences between the myosin IC isoforms and are an important step toward our understanding of the underlying mechanisms that regulate the various and distinct functions of myosin IC isoforms.

  12. GABAB(1) receptor subunit isoforms differentially regulate stress resilience

    Science.gov (United States)

    O’Leary, Olivia F.; Felice, Daniela; Galimberti, Stefano; Savignac, Hélène M.; Bravo, Javier A.; Crowley, Tadhg; El Yacoubi, Malika; Vaugeois, Jean-Marie; Gassmann, Martin; Bettler, Bernhard; Dinan, Timothy G.; Cryan, John F.

    2014-01-01

    Stressful life events increase the susceptibility to developing psychiatric disorders such as depression; however, many individuals are resilient to such negative effects of stress. Determining the neurobiology underlying this resilience is instrumental to the development of novel and more effective treatments for stress-related psychiatric disorders. GABAB receptors are emerging therapeutic targets for the treatment of stress-related disorders such as depression. These receptors are predominantly expressed as heterodimers of a GABAB(2) subunit with either a GABAB(1a) or a GABAB(1b) subunit. Here we show that mice lacking the GABAB(1b) receptor isoform are more resilient to both early-life stress and chronic psychosocial stress in adulthood, whereas mice lacking GABAB(1a) receptors are more susceptible to stress-induced anhedonia and social avoidance compared with wild-type mice. In addition, increased hippocampal expression of the GABAB(1b) receptor subunit is associated with a depression-like phenotype in the helpless H/Rouen genetic mouse model of depression. Stress resilience in GABAB(1b)−/− mice is coupled with increased proliferation and survival of newly born cells in the adult ventral hippocampus and increased stress-induced c-Fos activation in the hippocampus following early-life stress. Taken together, the data suggest that GABAB(1) receptor subunit isoforms differentially regulate the deleterious effects of stress and, thus, may be important therapeutic targets for the treatment of depression. PMID:25288769

  13. Entropy-based model for miRNA isoform analysis.

    Directory of Open Access Journals (Sweden)

    Shengqin Wang

    Full Text Available MiRNAs have been widely studied due to their important post-transcriptional regulatory roles in gene expression. Many reports have demonstrated the evidence of miRNA isoform products (isomiRs in high-throughput small RNA sequencing data. However, the biological function involved in these molecules is still not well investigated. Here, we developed a Shannon entropy-based model to estimate isomiR expression profiles of high-throughput small RNA sequencing data extracted from miRBase webserver. By using the Kolmogorov-Smirnov statistical test (KS test, we demonstrated that the 5p and 3p miRNAs present more variants than the single arm miRNAs. We also found that the isomiR variant, except the 3' isomiR variant, is strongly correlated with Minimum Free Energy (MFE of pre-miRNA, suggesting the intrinsic feature of pre-miRNA should be one of the important factors for the miRNA regulation. The functional enrichment analysis showed that the miRNAs with high variation, particularly the 5' end variation, are enriched in a set of critical functions, supporting these molecules should not be randomly produced. Our results provide a probabilistic framework for miRNA isoforms analysis, and give functional insights into pre-miRNA processing.

  14. Expression of aquaporin isoforms during human and mouse tooth development.

    Science.gov (United States)

    Felszeghy, S; Módis, L; Németh, P; Nagy, G; Zelles, T; Agre, P; Laurikkala, J; Fejerskov, O; Thesleff, I; Nielsen, S

    2004-04-01

    Previously, we described the development of hyaluronan (HA) deposition in human tooth germ tissues that are consistent with water transport in different stages of tooth development. The aquaporins (AQP) constitute a family of membrane water channels that are expressed in many organs. However, there are no data available about the expression pattern of aquaporin water channels in dental structures. In the present study we have characterised the expression of six different aquaporin isoforms (AQP1-5, AQP-9) in developing human and mouse tooth germs by immunohistochemistry using isoform specific antibodies. In the "bell stage" AQP1 was expressed in endothelial cells of small vessels whereas no other structures of the tooth primordial were labeled. AQP2, AQP3 and AQP9 immunoreactivity was not observed in tooth germs, whereas strong AQP4 and AQP5 expression was observed in dental lamina, inner enamel epithelium, stratum intermedium, stellate reticulum and the outer enamel epithelium. Oral epithelium also exhibited AQP4 and AQP5 immunolabeling. During development of the matrices of the dental hard tissues AQP4 and AQP5 immunostaining was observed in the odontoblasts and their processes, as well as in the secretory ameloblast and their apical processes. Immunolabeling controls were negative. In conclusion, AQP4 and AQP5 are expressed in tooth germ tissues in early development in cells that previously have been shown to express HA and/or CD44, indicating that AQP water channels may play a role for ECM hydration during tooth development.

  15. A Review of Metallothionein Isoforms and their Role in Pathophysiology

    Directory of Open Access Journals (Sweden)

    Senthil kumar M

    2011-05-01

    Full Text Available Abstract The Metallothionein (MT is a protein which has several interesting biological effects and has been demonstrated increase focus on the role of MT in various biological systems in the past three decades. The studies on the role of MT were limited with few areas like apoptosis and antioxidants in selected organs even fifty years after its discovery. Now acknowledge the exploration of various isoforms of MT such as MT-I, MT-II, MT-III and MT-IV and other isoforms in various biological systems. Strong evidence exists that MT modulates complex diseases and the immune system in the body but the primary function of MT still remains unknown. This review's main objective is to explore the capability to specifically manipulate MT levels in cells and in animals to provide answers regarding how MT could impact those complex disease scenarios. The experimental result mentioned in this review related among MT, zinc, cadmium, diabetic, heart disease, bone retardation, neuro toxicity, kidney dysfunction, cancer, and brain suggest novel method for exploration and contribute significantly to the growing scientist to research further in this field.

  16. Structures of alternatively spliced isoforms of human ketohexokinase.

    Science.gov (United States)

    Trinh, Chi H; Asipu, Aruna; Bonthron, David T; Phillips, Simon E V

    2009-03-01

    A molecular understanding of the unique aspects of dietary fructose metabolism may be the key to understanding and controlling the current epidemic of fructose-related obesity, diabetes and related adverse metabolic states in Western populations. Fructose catabolism is initiated by its phosphorylation to fructose 1-phosphate, which is performed by ketohexokinase (KHK). Here, the crystal structures of the two alternatively spliced isoforms of human ketohexokinase, hepatic KHK-C and the peripheral isoform KHK-A, and of the ternary complex of KHK-A with the substrate fructose and AMP-PNP are reported. The structure of the KHK-A ternary complex revealed an active site with both the substrate fructose and the ATP analogue in positions ready for phosphorylation following a reaction mechanism similar to that of the pfkB family of carbohydrate kinases. Hepatic KHK deficiency causes the benign disorder essential fructosuria. The effects of the disease-causing mutations (Gly40Arg and Ala43Thr) have been modelled in the context of the KHK structure.

  17. BDNF isoforms: a round trip ticket between neurogenesis and serotonin?

    Science.gov (United States)

    Foltran, Rocío Beatriz; Diaz, Silvina Laura

    2016-07-01

    The brain-derived neurotrophic factor, BDNF, was discovered more than 30 years ago and, like other members of the neurotrophin family, this neuropeptide is synthetized as a proneurotrophin, the pro-BDNF, which is further cleaved to yield mature BDNF. The myriad of actions of these two BDNF isoforms in the central nervous system is constantly increasing and requires the development of sophisticated tools and animal models to refine our understanding. This review is focused on BDNF isoforms, their participation in the process of neurogenesis taking place in the hippocampus of adult mammals, and the modulation of their expression by serotonergic agents. Interestingly, around this triumvirate of BDNF, serotonin, and neurogenesis, a series of recent research has emerged with apparently counterintuitive results. This calls for an exhaustive analysis of the data published so far and encourages thorough work in the quest for new hypotheses in the field. BDNF is synthetized as a pre-proneurotrophin. After removal of the pre-region, proBDNF can be cleaved by intracellular or extracellular proteases. Mature BDNF can bind TrkB receptors, promoting their homodimerization and intracellular phosphorylation. Phosphorylated-TrkB can activate three different signaling pathways. Whereas G-protein-coupled receptors can transactivate TrkB receptors, truncated forms can inhibit mBDNF signaling. Pro-BDNF binds p75(NTR) by its mature domain, whereas the pro-region binds co-receptors. © 2016 International Society for Neurochemistry.

  18. Development of a Microwave Regenerative Sorbent-Based Hydrogen Purifier

    Science.gov (United States)

    Wheeler, Richard R., Jr.; Dewberry, Ross H.; McCurry, Bryan D.; Abney, Morgan B.; Greenwood, Zachary W.

    2016-01-01

    This paper describes the design and fabrication of a Microwave Regenerative Sorbent-based Hydrogen Purifier (MRSHP). This unique microwave powered technology was developed for the purification of a hydrogen stream produced by the Plasma Pyrolysis Assembly (PPA). The PPA is a hydrogen recovery (from methane) post processor for NASA's Sabatier-based carbon dioxide reduction process. Embodied in the Carbon dioxide Reduction Assembly (CRA), currently aboard the International Space Station (ISS), the Sabatier reaction employs hydrogen to catalytically recover oxygen, in the form of water, from respiratory carbon dioxide produced by the crew. This same approach is base-lined for future service in the Air Revitalization system on extended missions into deep space where resupply is not practical. Accordingly, manned exploration to Mars may only become feasible with further closure of the air loop as afforded by the greater hydrogen recovery permitted by the PPA with subsequent hydrogen purification. By utilizing the well-known high sorbate loading capacity of molecular sieve 13x, coupled with microwave dielectric heating phenomenon, MRSHP technology is employed as a regenerative filter for a contaminated hydrogen gas stream. By design, freshly regenerated molecular sieve 13x contained in the MRSHP will remove contaminants from the effluent of a 1-CM scale PPA for several hours prior to breakthrough. By reversing flow and pulling a relative vacuum the MRSHP prototype then uses 2.45 GHz microwave power, applied through a novel coaxial antenna array, to rapidly heat the sorbent bed and drive off the contaminants in a short duration vacuum/thermal contaminant desorption step. Finally, following rapid cooling via room temperature cold plates, the MRSHP is again ready to serve as a hydrogen filter.

  19. Correcting for purifying selection: an improved human mitochondrial molecular clock.

    Science.gov (United States)

    Soares, Pedro; Ermini, Luca; Thomson, Noel; Mormina, Maru; Rito, Teresa; Röhl, Arne; Salas, Antonio; Oppenheimer, Stephen; Macaulay, Vincent; Richards, Martin B

    2009-06-01

    There is currently no calibration available for the whole human mtDNA genome, incorporating both coding and control regions. Furthermore, as several authors have pointed out recently, linear molecular clocks that incorporate selectable characters are in any case problematic. We here confirm a modest effect of purifying selection on the mtDNA coding region and propose an improved molecular clock for dating human mtDNA, based on a worldwide phylogeny of > 2000 complete mtDNA genomes and calibrating against recent evidence for the divergence time of humans and chimpanzees. We focus on a time-dependent mutation rate based on the entire mtDNA genome and supported by a neutral clock based on synonymous mutations alone. We show that the corrected rate is further corroborated by archaeological dating for the settlement of the Canary Islands and Remote Oceania and also, given certain phylogeographic assumptions, by the timing of the first modern human settlement of Europe and resettlement after the Last Glacial Maximum. The corrected rate yields an age of modern human expansion in the Americas at approximately 15 kya that-unlike the uncorrected clock-matches the archaeological evidence, but continues to indicate an out-of-Africa dispersal at around 55-70 kya, 5-20 ky before any clear archaeological record, suggesting the need for archaeological research efforts focusing on this time window. We also present improved rates for the mtDNA control region, and the first comprehensive estimates of positional mutation rates for human mtDNA, which are essential for defining mutation models in phylogenetic analyses.

  20. Positive and purifying selection on the Drosophila Y chromosome.

    Science.gov (United States)

    Singh, Nadia D; Koerich, Leonardo B; Carvalho, Antonio Bernardo; Clark, Andrew G

    2014-10-01

    Y chromosomes, with their reduced effective population size, lack of recombination, and male-limited transmission, present a unique collection of constraints for the operation of natural selection. Male-limited transmission may greatly increase the efficacy of selection for male-beneficial mutations, but the reduced effective size also inflates the role of random genetic drift. Together, these defining features of the Y chromosome are expected to influence rates and patterns of molecular evolution on the Y as compared with X-linked or autosomal loci. Here, we use sequence data from 11 genes in 9 Drosophila species to gain insight into the efficacy of natural selection on the Drosophila Y relative to the rest of the genome. Drosophila is an ideal system for assessing the consequences of Y-linkage for molecular evolution in part because the gene content of Drosophila Y chromosomes is highly dynamic, with orthologous genes being Y-linked in some species whereas autosomal in others. Our results confirm the expectation that the efficacy of natural selection at weakly selected sites is reduced on the Y chromosome. In contrast, purifying selection on the Y chromosome for strongly deleterious mutations does not appear to be compromised. Finally, we find evidence of recurrent positive selection for 4 of the 11 genes studied here. Our results thus highlight the variable nature of the mode and impact of natural selection on the Drosophila Y chromosome. © The Author 2014. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Fat lowers fat: purified phospholipids as emerging therapies for dyslipidemia.

    Science.gov (United States)

    Sahebkar, Amirhossein

    2013-04-01

    Dyslipidemia is a major coronary heart disease (CHD) risk factor. In spite of the proven efficacy of statin drugs in reducing CHD burden, there is still much room for the discovery of novel therapeutic agents to address the considerable residual cardiovascular risk that remains after treatment with currently available medications. In particular, there is an urgent demand for drugs capable of boosting the concentration and/or function of high-density lipoprotein (HDL) and apolipoprotein A-I (apo A-I), thereby promoting reverse cholesterol transport. Phospholipids are naturally occurring fats that play indispensible role in human health via their structural, energy storage, signal transduction and metabolic functions. Supplementation with either purified or mixed preparations of bioactive phospholipids has been reported to ameliorate a range of nutritional and cardiovascular disorders. Moreover, several lines of evidence have supported the efficacy of dietary phospholipids in reducing serum and hepatic contents of cholesterol and triglycerides, while increasing HDL-C and apo A-I levels. These beneficial effects of phospholipids could be attributed to their ability in reducing intestinal cholesterol absorption, enhancing biliary cholesterol excretion and modulating the expression and activity of transcriptional factors and enzymes that are involved in lipoprotein metabolism. Given their extreme safety and biocompatibility, dietary supplementation with phospholipid preparations, in particular phosphatidylinositol, appears as a novel and effective strategy that could be used as an alternative or adjunctive therapy to the current medications. The present review outlines the in-vitro, in-vivo and clinical findings on the anti-dyslipidemic effects of three most abundant phospholipids in the human body and diet namely phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol.

  2. Purifier-integrated methanol reformer for fuel cell vehicles

    Science.gov (United States)

    Han, Jaesung; Kim, Il-soo; Choi, Keun-Sup

    We developed a compact, 3-kW, purifier-integrated modular reformer which becomes the building block of full-scale 30-kW or 50-kW methanol fuel processors for fuel cell vehicles. Our proprietary technologies regarding hydrogen purification by composite metal membrane and catalytic combustion by washcoated wire-mesh catalyst were combined with the conventional methanol steam-reforming technology, resulting in higher conversion, excellent quality of product hydrogen, and better thermal efficiency than any other systems using preferential oxidation. In this system, steam reforming, hydrogen purification, and catalytic combustion all take place in a single reactor so that the whole system is compact and easy to operate. Hydrogen from the module is ultrahigh pure (99.9999% or better), hence there is no power degradation of PEMFC stack due to contamination by CO. Also, since only pure hydrogen is supplied to the anode of the PEMFC stack, 100% hydrogen utilization is possible in the stack. The module produces 2.3 Nm 3/h of hydrogen, which is equivalent to 3 kW when PEMFC has 43% efficiency. Thermal efficiency (HHV of product H 2/HHV of MeOH in) of the module is 89% and the power density of the module is 0.77 kW/l. This work was conducted in cooperation with Hyundai Motor Company in the form of a Korean national project. Currently the module is under test with an actual fuel cell stack in order to verify its performance. Sooner or later a full-scale 30-kW system will be constructed by connecting these modules in series and parallel and will serve as the fuel processor for the Korean first fuel cell hybrid vehicle.

  3. A close-up of the ALEPH Time Projection Chamber (TPC) after the detector was dismantled, with a slice removed for the exhibition at the Musée International d'Horlogerie.

    CERN Multimedia

    2004-01-01

    On the occasion of CERN's 50th anniversary, starting 2 December at the Musée International d'Horlogerie in La Chaux-de-Fonds, Switzerland, a new exhibition will pay tribute to physics, from the cosmic to the subatomic scales. The exhibition, which run for several years, includes a slice of the Time Projection Chamber (TPC) from ALEPH, one of the LEP detectors, which CERN donated to the museum when the detector was dismantled.

  4. ArgonCube: a novel, fully-modular approach for the realization of large-mass liquid argon TPC neutrino detectors

    CERN Document Server

    Amsler, C; Asaadi, J; Auger, M; Barbato, F; Bay, F; Bishai, M; Bleiner, D; Borgschulte, A; Bremer, J; Cavus, E; Chen, H; De Geronimo, G; Ereditato, A; Fleming, B; Goldi, D; Hanni, R; Kose, U; Kreslo, I; La Mattina, F; Lanni, F; Lissauer, D; Luthi, M; Lutz, P; Marchionni, A; Mladenov, D; Nessi, M; Noto, F; Palamara, O; Raaf, J L; Radeka, V; Rudolph Von Rohr, Ch; Smargianaki, D; Soderberg, M; Strauss, Th; Weber, M; Yu, B; Zeller, G P; Zeyrek, M; CERN. Geneva. SPS and PS Experiments Committee; SPSC

    2015-01-01

    The Liquid Argon Time Projection Chamber is a prime candidate detector for future neutrino oscillation physics experiments, underground neutrino observatories and proton decay searches. A large international project based on this technology is currently being considered at the future LBNF facility in the United States on the very large mass scale of 40 kton. In this document, following the long standing R&D work conducted over the last years in several laboratories in Europe and in the United States, we intend to propose a novel Liquid Argon TPC approach based on a fully-modular, innovative design, the ArgonCube. The related R&D work will proceed along two main directions; one aimed at on the assessment of the proposed modular detector design, the other on the exploitation of new signal readout methods. Such a strategy will provide high performance while being cost-effective and robust at the same time. According to our plans, we will firstly realize a detector prototype hosted in a cryostat that is a...

  5. Determination of Muon Momentum in the MicroBooNE LArTPC Using an Improved Model of Multiple Coulomb Scattering

    Energy Technology Data Exchange (ETDEWEB)

    Abratenko, P.; et al.

    2017-03-17

    We discuss a technique for measuring a charged particle's momentum by means of multiple Coulomb scattering (MCS) in the MicroBooNE liquid argon time projection chamber (LArTPC). This method does not require the full particle ionization track to be contained inside of the detector volume as other track momentum reconstruction methods do (range-based momentum reconstruction and calorimetric momentum reconstruction). We motivate use of this technique, describe a tuning of the underlying phenomenological formula, quantify its performance on fully contained beam-neutrino-induced muon tracks both in simulation and in data, and quantify its performance on exiting muon tracks in simulation. We find agreement between data and simulation for contained tracks, with a small bias in the momentum reconstruction and with resolutions that vary as a function of track length, improving from about 10% for the shortest (one meter long) tracks to 5% for longer (several meter) tracks. For simulated exiting muons with at least one meter of track contained, we find a similarly small bias, and a resolution which is less than 15% for muons with momentum below 2 GeV/c.

  6. Expression of Interest for a Full-Scale Detector Engineering Test and Test Beam Calibration of a Single-Phase LAr TPC

    CERN Document Server

    Leigui de Oliveira, M A

    2014-01-01

    Following the recommendation of the U.S. Particle Physics Project Prioritization Panel[1], Fermilab is working with the world neutrino community, CERN, and others to establish “a new international collaboration to design and execute a highly capable Long-Baseline Neutrino Facility (LBNF) hosted by the U.S.” This new collaboration, which is expected to be formed during the next several months, will combine, among others, groups that have been developing both single-phase and dual-phase LAr TPC detectors for long-baseline physics, mainly from the LBNE and LBNO collaborations respectively. This Expression of Interest regards a proposed full-scale prototype and beam test of the LBNE-design single-phase detector, utilizing the CERN Neutrino Platform[2] that was recently approved as part of the Medium-Term Plan (MTP) [3]. Once the LBNF collaboration is formed, it is expected that development of both single- and dual-phase detectors will come under the umbrella of the new collaboration.

  7. Single-phase ProtoDUNE, the Prototype of a Single-Phase Liquid Argon TPC for DUNE at the CERN Neutrino Platform

    CERN Document Server

    Cavanna, F; Touramanis, C

    2017-01-01

    ProtoDUNE-SP is the single-phase DUNE Far Detector prototype that is under construction and will be operated at the CERN Neutrino Platform (NP) starting in 2018. It was proposed to the CERN SPSC in June 2015 (SPSC-P-351) and was approved in December 2015 as experiment NP04 (ProtoDUNE). ProtoDUNE-SP, a crucial part of the DUNE effort towards the construction of the first DUNE 10-kt fiducial mass far detector module (17 kt total LAr mass), is a significant experiment in its own right. With a total liquid argon (LAr) mass of 0.77 kt, it represents the largest monolithic single phase LArTPC detector to be built to date. It is housed in an extension to the EHN1 hall in the North Area, where the CERN NP is providing a new dedicated charged-particle test beamline. ProtoDUNE-SP aims to take its first beam data before the LHC long shutdown (LS2) at the end of 2018. ProtoDUNE-SP prototypes the designs of most of the single-phase DUNE far detector module (DUNE-SP) components at a 1:1 scale, with an extrapolation of abo...

  8. Near-intrinsic energy resolution for 30–662 keV gamma rays in a high pressure xenon electroluminescent TPC

    Energy Technology Data Exchange (ETDEWEB)

    Álvarez, V. [Instituto de Física Corpuscular (IFIC), CSIC and Universitat de València, Calle Catedrático José Beltrán, 2, 46980 Paterna, Valencia (Spain); Borges, F.I.G.M. [Departamento de Fisica, Universidade de Coimbra, Rua Larga, 3004-516 Coimbra (Portugal); Cárcel, S. [Instituto de Física Corpuscular (IFIC), CSIC and Universitat de València, Calle Catedrático José Beltrán, 2, 46980 Paterna, Valencia (Spain); Castel, J.; Cebrián, S. [Laboratorio de Física Nuclear y Astropartículas, Universidad de Zaragoza, Calle Pedro Cerbuna 12, 50009 Zaragoza (Spain); Cervera, A. [Instituto de Física Corpuscular (IFIC), CSIC and Universitat de València, Calle Catedrático José Beltrán, 2, 46980 Paterna, Valencia (Spain); Conde, C.A.N. [Departamento de Fisica, Universidade de Coimbra, Rua Larga, 3004-516 Coimbra (Portugal); Dafni, T. [Laboratorio de Física Nuclear y Astropartículas, Universidad de Zaragoza, Calle Pedro Cerbuna 12, 50009 Zaragoza (Spain); Dias, T.H.V.T. [Departamento de Fisica, Universidade de Coimbra, Rua Larga, 3004-516 Coimbra (Portugal); Díaz, J. [Instituto de Física Corpuscular (IFIC), CSIC and Universitat de València, Calle Catedrático José Beltrán, 2, 46980 Paterna, Valencia (Spain); and others

    2013-04-21

    We present the design, data and results from the NEXT prototype for Double Beta and Dark Matter (NEXT-DBDM) detector, a high-pressure gaseous natural xenon electroluminescent time projection chamber (TPC) that was built at the Lawrence Berkeley National Laboratory. It is a prototype of the planned NEXT-100 {sup 136}Xe neutrino-less double beta decay (0νββ) experiment with the main objectives of demonstrating near-intrinsic energy resolution at energies up to 662 keV and of optimizing the NEXT-100 detector design and operating parameters. Energy resolutions of ∼1% FWHM for 662 keV gamma rays were obtained at 10 and 15 atm and ∼5% FWHM for 30 keV fluorescence xenon X-rays. These results demonstrate that 0.5% FWHM resolutions for the 2459 keV hypothetical neutrino-less double beta decay peak are realizable. This energy resolution is a factor 7–20 better than that of the current leading 0νββ experiments using liquid xenon and thus represents a significant advancement. We present also first results from a track imaging system consisting of 64 silicon photo-multipliers recently installed in NEXT–DBDM that, along with the excellent energy resolution, demonstrates the key functionalities required for the NEXT-100 0νββ search.

  9. First measurement of polarisation asymmetry of a gamma-ray beam between 1.74 to 74 MeV with the HARPO TPC

    CERN Document Server

    Gros, Philippe; Attié, David; Bernard, Denis; Bruel, Philippe; Calvet, Denis; Colas, Paul; Daté, Schin; Delbart, Alain; Frotin, Mickael; Geerebaert, Yannick; Giebels, Berrie; Götz, Diego; Hashimoto, S; Horan, Deirdr; Kotaka, T; Louzir, Marc; Minamiyama, Y; Miyamoto, Shuji; Ohkuma, H; Poilleux, Patrick; Semeniouk, Igor; Sizun, Patrick; Takemoto, A; Yamaguchi, M; Wang, Shaobo

    2016-01-01

    Current $\\gamma$-ray telescopes suffer from a gap in sensitivity in the energy range between 100keV and 100MeV, and no polarisation measurement has ever been done on cosmic sources above 1MeV. Past and present e$^+$e$^-$ pair telescopes are limited at lower energies by the multiple scattering of electrons in passive tungsten converter plates. This results in low angular resolution, and, consequently, a drop in sensitivity to point sources below 1GeV. The polarisation information, which is carried by the azimuthal angle of the conversion plane, is lost for the same reasons. HARPO (Hermetic ARgon POlarimeter) is an R\\&D program to characterise the operation of a gaseous detector (a Time Projection Chamber or TPC) as a high angular-resolution and sensitivity telescope and polarimeter for $\\gamma$ rays from cosmic sources. It represents a first step towards a future space instrument in the MeV-GeV range. We built and characterised a 30cm cubic demonstrator [SPIE 91441M], and put it in a polarised $\\gamma$-ray...

  10. Differential sensitivity of rat voltage-sensitive sodium channel isoforms to pyrazoline-type insecticides.

    Science.gov (United States)

    Silver, Kristopher S; Soderlund, David M

    2006-07-15

    Pyrazoline-type insecticides are potent inhibitors of insect and mammalian voltage-sensitive sodium channels. In mammals, there are nine sodium channel alpha subunit isoforms that have unique distributions and pharmacological properties, but no published data exist that compare the relative sensitivity of these different mammalian sodium channel isoforms to inhibition by pyrazoline-type insecticides. This study employed the Xenopus oocyte expression system to examine the relative sensitivity of rat Na(v)1.2a, Na(v)1.4, Na(v)1.5, and Na(v)1.8 sodium channel alpha subunit isoforms to the pyrazoline-type insecticides indoxacarb, DCJW, and RH 3421. Additionally, we assessed the effect of coexpression with the rat beta1 auxiliary subunit on the sensitivity of the Na(v)1.2a and Na(v)1.4 isoforms to these compounds. The relative sensitivity of the four sodium channel alpha subunits differed for each of the three compounds we examined. With DCJW, the order of sensitivity was Na(v)1.4 > Na(v)1.2a > Na(v)1.5 > Na(v)1.8. In contrast, the relative sensitivity of these isoforms to indoxacarb differed from that to DCJW: the Na(v)1.8 isoform was most sensitive, the Na(v)1.4 isoform was completely insensitive, and the sensitivities of the Na(v)1.5 and Na(v)1.2a isoforms were intermediate between these two extremes. Moreover, the pattern of sensitivity to RH 3421 among these four isoforms was different from that for either indoxacarb or DCJW: the Na(v)1.4 isoform was most sensitive to RH 3421, whereas the sensitivities of the remaining three isoforms were substantially less than that of the Na(v)1.4 isoform and were approximately equivalent. The only statistically significant effect of coexpression of either the Na(v)1.2a or Na(v)1.4 isoforms with the beta1 subunit was the modest reduction in the sensitivity of the Na(v)1.2a isoform to RH 3421. These results demonstrate that mammalian sodium channel isoforms differ in their sensitivities to pyrazoline-type insecticides.

  11. Nesprins: tissue-specific expression of epsilon and other short isoforms.

    Directory of Open Access Journals (Sweden)

    Nguyen Thuy Duong

    Full Text Available Nesprin-1-giant and nesprin-2-giant regulate nuclear positioning by the interaction of their C-terminal KASH domains with nuclear membrane SUN proteins and their N-terminal calponin-homology domains with cytoskeletal actin. A number of short isoforms lacking the actin-binding domains are produced by internal promotion. We have evaluated the significance of these shorter isoforms using quantitative RT-PCR and western blotting with site-specific monoclonal antibodies. Within a complete map of nesprin isoforms, we describe two novel nesprin-2 epsilon isoforms for the first time. Epsilon isoforms are similar in size and structure to nesprin-1-alpha. Expression of nesprin isoforms was highly tissue-dependent. Nesprin-2-epsilon-1 was found in early embryonic cells, while nesprin-2-epsilon-2 was present in heart and other adult tissues, but not skeletal muscle. Some cell lines lack shorter isoforms and express only one of the two nesprin genes, suggesting that either of the giant nesprins is sufficient for basic cell functions. For the first time, localisation of endogenous nesprin away from the nuclear membrane was shown in cells where removal of the KASH domain by alternative splicing occurs. By distinguishing between degradation products and true isoforms on western blots, it was found that previously-described beta and gamma isoforms are expressed either at only low levels or with a limited tissue distribution. Two of the shortest alpha isoforms, nesprin-1-alpha-2 and nesprin-2-alpha-1, were found almost exclusively in cardiac and skeletal muscle and a highly conserved and alternatively-spliced exon, available in both nesprin genes, was always included in these tissues. These "muscle-specific" isoforms are thought to form a complex with emerin and lamin A/C at the inner nuclear membrane and mutations in all three proteins cause Emery-Dreifuss muscular dystrophy and/or inherited dilated cardiomyopathy, disorders in which only skeletal muscle and

  12. Expression of phosphoinositide-specific phospholipase C isoforms in native endothelial cells.

    Directory of Open Access Journals (Sweden)

    Delphine M Béziau

    Full Text Available Phospholipase C (PLC comprises a superfamily of enzymes that play a key role in a wide array of intracellular signalling pathways, including protein kinase C and intracellular calcium. Thirteen different mammalian PLC isoforms have been identified and classified into 6 families (PLC-β, γ, δ, ε, ζ and η based on their biochemical properties. Although the expression of PLC isoforms is tissue-specific, concomitant expression of different PLC has been reported, suggesting that PLC family is involved in multiple cellular functions. Despite their critical role, the PLC isoforms expressed in native endothelial cells (ECs remains undetermined. A conventional PCR approach was initially used to elucidate the mRNA expression pattern of PLC isoforms in 3 distinct murine vascular beds: mesenteric (MA, pulmonary (PA and middle cerebral arteries (MCA. mRNA encoding for most PLC isoforms was detected in MA, MCA and PA with the exception of η2 and β2 (only expressed in PA, δ4 (only expressed in MCA, η1 (expressed in all but MA and ζ (not detected in any vascular beds tested. The endothelial-specific PLC expression was then sought in freshly isolated ECs. Interestingly, the PLC expression profile appears to differ across the investigated arterial beds. While mRNA for 8 of the 13 PLC isoforms was detected in ECs from MA, two additional PLC isoforms were detected in ECs from PA and MCA. Co-expression of multiple PLC isoforms in ECs suggests an elaborate network of signalling pathways: PLC isoforms may contribute to the complexity or diversity of signalling by their selective localization in cellular microdomains. However in situ immunofluorescence revealed a homogeneous distribution for all PLC isoforms probed (β3, γ2 and δ1 in intact endothelium. Although PLC isoforms play a crucial role in endothelial signal transduction, subcellular localization alone does not appear to be sufficient to determine the role of PLC in the signalling microdomains found

  13. Optimization of freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel

    National Research Council Canada - National Science Library

    Mehrnoush, Amid; Mustafa, Shuhaimi; Yazid, Abdul Manap Mohd

    2012-01-01

    Response surface methodology (RSM) along with central composite design (CCD) was applied to optimize the freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel...

  14. Hypolipidemic activities of xanthorrhizol purified from centrifugal TLC.

    Science.gov (United States)

    Oon, Seok Fang; Nallappan, Meenakshii; Kassim, Nur Kartinee; Shohaimi, Shamarina; Sa'ariwijaya, Mohd Shazrul Fazry; Tee, Thiam Tsui; Cheah, Yew Hoong

    2016-09-23

    Hyperlipidemia is defined as the presence of either hypertriglyceridemia or hypercholesterolemia, which could cause atherosclerosis. Although hyperlipidemia can be treated by hypolipidemic drugs, they are limited due to lack of effectiveness and safety. Previous studies demonstrated that xanthorrhizol (XNT) isolated from Curcuma xanthorrhizza Roxb. reduced the levels of free fatty acid and triglyceride in vivo. However, its ability to inhibit cholesterol uptake in HT29 colon cells and adipogenesis in 3T3-L1 cells are yet to be reported. In this study, XNT purified from centrifugal TLC demonstrated 98.3% purity, indicating it could be an alternative purification method. The IC50 values of XNT were 30.81 ± 0.78 μg/mL in HT29 cells and 35.07 ± 0.24 μg/mL in 3T3-L1 adipocytes, respectively. Cholesterol uptake inhibition study using HT29 colon cells showed that XNT (15 μg/mL) significantly inhibited the fluorescent cholesterol analogue NBD uptake by up to 27 ± 3.1% relative to control. On the other hand, higher concentration of XNT (50 μg/mL) significantly suppressed the growth of 3T3-L1 adipocytes (5.9 ± 0.58%) compared to 3T3-L1 preadipocytes (81.31 ± 0.55%). XNT was found to impede adipogenesis of 3T3-L1 adipocytes in a dose-dependent manner from 3.125 to 12.5 μg/mL, where 12.5 μg/mL significantly suppressed 36.13 ± 2.1% of lipid accumulation. We postulate that inhibition of cholesterol uptake, adipogenesis, preadipocyte and adipocyte number may be utilized as treatment modalities to reduce the prevalence of lipidemia. To conclude, XNT could be a potential hypolipidemic agent to improve cardiovascular health in the future. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  15. Properties of serine: glyoxylate aminotransferase purified from Arabidopsis thaliana leaves

    Institute of Scientific and Technical Information of China (English)

    Maria Kendziorek; Andrzej Paszkowski

    2008-01-01

    The photorespiratory enzyme L-serine: glyoxylate aminotransferase (SGAT; EC 2.6.1.45) was purified from Arabidopsis thaliana leaves. The final enzyme was approximately 80% pure as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with silver staining. The identity of the enzyme was confirmed by LC/MS/MS analysis.The molecular mass estimated by gel filtration chromatography on Sephadex G-150 under non-denaturing conditions, mass spectrometry (matrix-assisted laser desorption/ionization/time of flight technique) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 82.4 kDa,42.0 kDa, and 39.8 kDa, respectively, indicating dimer as the active form. The optimum Ph value was 9.2. The enzyme activity was inhibited by aminooxyacetate and β-chloro-L-alanine both compounds reacting with the carbonyl group of pyridoxal phosphate. The enzyme's transaminating activity with L-alanine and glyoxylate as substrates was approximately 55% of that observed with L-serine and glyoxylate, The lower Km value (1.25 Mm) for L-alanine, compared with that of other plant SGATs, and the kcat/Km(Ala) ratio being approximately 2-fold higher than kcat/Km(Ser) suggested that, during photorespiration, Ala and Ser are used by Arabidopsis SGAT with equal efficiency as amino group donors for glyoxylate. The equilibrium constant (Keq), derived from the Haldane relation, for the transamination reaction between L-serine and glyoxylate with the formation of hydroxypyruvate and glycine was 79.1, strongly favoring glycine synthesis. However, it was accompanied by a low Km value of 2.83 Mm for glycine. A comparison of some kinetic properties of the studied enzymes with the recombinant Arabidopsis SGATs previously obtained revealed substantial differences. The ratio of the velocity of the transamination reaction with L-alanine and glyoxylate as substrates versus that with L-serine and glyoxylate was 1:1.8 for the native enzyme, whereas it was 1: 7 for the recombinant SGAT

  16. [Effects of hydroxyl radicals on purified angiotensin I converting enzyme].

    Science.gov (United States)

    Michel, B; Nirina, L B; Grima, M; Ingert, C; Coquard, C; Barthelmebs, M; Imbs, J L

    1998-08-01

    Somatic angiotensin-converting enzyme (ACE) is a protein which contains two similar domains (N and C), each possessing a functional active site. The relationship between ACE, its natural substrates and oxygen free radicals is starting to be explored. On one hand, superoxide anions production is induced by angiotensin II and on the other hand, activated polynuclear neutrophils, through free radicals generation, alter endothelial ACE activity. In this study, we examined the impact of hydroxyl radicals (.OH) on purified ACE. .OH were produced using a generator: 2,2'-azo-bis 2-amidinopropane (GRH) provided by Lara-Spiral (Fr). GRH (3 mM), in a time-dependent fashion, inhibited ACE activity. When ACE was co-incubated for 4 h with GRH, its activity decreased by 70%. Addition of dimethylthiourea (DMTU: 0.03 to 1 mM) or mannitol + methionine (20/10 mM), two sets of .OH scavengers, produced a dose-dependent protection on ACE activity. To examine whether oxidation of thiol groups in the ACE molecule could be involved in the action of GRH, the effects of thiol reducing agents: mercaptoethanol and dithiotreitol (DTT) were investigated. These compounds produced a dose-dependent and significant protection; with 100% protection at 0.2 and 0.3 mM for mercaptoethanol and at 0.1 mM for DTT. The hydrolysis of two natural and domain-specific substrates were also explored. The hydrolysis of angiotensin I preferentially cleaved by the C domain was significantly (p GRH [in nmol angio II formed/min/nmol of ACE, n = 4; 35.9 +/- 0.6 (control), 15.5 +/- 2.8 (GRH : 0.3 mM), 15.1 +/- 0.5 (1), 10.9 +/- 0.6 (3)]. The hydrolysis of the hemoregulatory peptide (hp), preferential substrate for the N domain was not affected by GRH at 0.3 mM and inhibited by 28% (not significant) by 1 mM GRH [in nmol ph hydrolized/min/nmol ACE, n = 4; 12.6 +/- 1.9 (control), 14.9 (GRH : 0.3 mM), 8.3 +/- 4.0 (1). These results demonstrated that .OH affect ACE activity and could suggest a privileged impact of GRH on the

  17. Different phosphoinositide 3-kinase isoforms mediate carrageenan nociception and inflammation.

    Science.gov (United States)

    Pritchard, Rory A; Falk, Lovissa; Larsson, Mathilda; Leinders, Mathias; Sorkin, Linda S

    2016-01-01

    Phosphoinositide 3-kinases (PI3Ks) participate in signal transduction cascades that can directly activate and sensitize nociceptors and enhance pain transmission. They also play essential roles in chemotaxis and immune cell infiltration leading to inflammation. We wished to determine which PI3K isoforms were involved in each of these processes. Lightly anesthetized rats (isoflurane) were injected subcutaneously with carrageenan in their hind paws. This was preceded by a local injection of 1% DMSO vehicle or an isoform-specific antagonist to PI3K-α (compound 15-e), -β (TGX221), -δ (Cal-101), or -γ (AS252424). We measured changes in the mechanical pain threshold and spinal c-Fos expression (4 hours after injection) as indices of nociception. Paw volume, plasma extravasation (Evans blue, 0.3 hours after injection), and neutrophil (myeloperoxidase; 1 hour after injection) and macrophage (CD11b+; 4 hour after injection) infiltration into paw tissue were the measured inflammation endpoints. Only PI3K-γ antagonist before treatment reduced the carrageenan-induced pain behavior and spinal expression of c-Fos (P ≤ 0.01). In contrast, pretreatment with PI3K-α, -δ, and-γ antagonists reduced early indices of inflammation. Plasma extravasation PI3K-α (P ≤ 0.05), -δ (P ≤ 0.05), and -γ (P ≤ 0.01), early (0-2 hour) edema -α (P ≤ 0.05), -δ (P ≤ 0.001), and -γ (P ≤ 0.05), and neutrophil infiltration (all P ≤ 0.001) were all reduced compared to vehicle pretreatment. Later (2-4 hour), edema and macrophage infiltration (P ≤ 0.05) were reduced by only the PI3K-δ and -γ isoform antagonists, with the PI3K-δ antagonist having a greater effect on edema. PI3K-β antagonism was ineffective in all paradigms. These data indicate that pain and clinical inflammation are pharmacologically separable and may help to explain clinical conditions in which inflammation naturally wanes or goes into remission, but pain continues unabated.

  18. Crystallization and preliminary X-ray diffraction studies of NP24-I, an isoform of a thaumatin-like protein from ripe tomato fruits

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, Raka; Chakrabarti, Chandana, E-mail: chandana.chakrabarti@saha.ac.in [Crystallography and Molecular Biology Division, Saha Institute of Nuclear Physics, Kolkata 700064 (India)

    2005-08-01

    A thaumatin-like antifungal protein, NP24-I, has been isolated from ripe tomato fruits. It was crystallized by the vapour-diffusion method and data were collected to 2.45 Å. The structure was solved by molecular replacement. NP24 is a 24 kDa (207-amino-acid) antifungal thaumatin-like protein (TLP) found in tomato fruits. An isoform of the protein, NP24-I, is reported to play a possible role in ripening of the fruit in addition to its antifungal properties. The protein has been isolated and purified and crystallized by the hanging-drop vapour-diffusion method. The crystals belong to the tetragonal space group P4{sub 3}, with unit-cell parameters a = b = 61.01, c = 62.90 Å and one molecule per asymmetric unit. X-ray diffraction data were processed to a resolution of 2.45 Å and the structure was solved by molecular replacement.

  19. Insulin receptor isoforms : an integrated view focused on gestational diabetes mellitus

    NARCIS (Netherlands)

    Westermeier, F.; Sáez, T.; Arroyo, P.; Toledo, F.; Gutierrez, J.; Sanhueza, C.; Pardo, F.; Leiva, A.; Sobrevia, L.

    2016-01-01

    The human insulin receptor (IR) exists in two isoforms that differ by the absence (IR-A) or the presence (IR-B) of a 12-amino acid segment encoded by exon 11. Both isoforms are functionally distinct regarding their binding affinities and intracellular signalling. However, the underlying mechanisms r

  20. Glial fibrillary acidic protein isoform expression in plaque related astrogliosis in Alzheimer's disease.

    Science.gov (United States)

    Kamphuis, Willem; Middeldorp, Jinte; Kooijman, Lieneke; Sluijs, Jacqueline A; Kooi, Evert-Jan; Moeton, Martina; Freriks, Michel; Mizee, Mark R; Hol, Elly M

    2014-03-01

    In Alzheimer's disease (AD), amyloid plaques are surrounded by reactive astrocytes with an increased expression of intermediate filaments including glial fibrillary acidic protein (GFAP). Different GFAP isoforms have been identified that are differentially expressed by specific subpopulations of astrocytes and that impose different properties to the intermediate filament network. We studied transcript levels and protein expression patterns of all known GFAP isoforms in human hippocampal AD tissue at different stages of the disease. Ten different transcripts for GFAP isoforms were detected at different abundancies. Transcript levels of most isoforms increased with AD progression. GFAPδ-immunopositive astrocytes were observed in subgranular zone, hilus, and stratum-lacunosum-moleculare. GFAPδ-positive cells also stained for GFAPα. In AD donors, astrocytes near plaques displayed increased staining of both GFAPα and GFAPδ. The reading-frame-shifted isoform, GFAP(+1), staining was confined to a subset of astrocytes with long processes, and their number increased in the course of AD. In conclusion, the various GFAP isoforms show differential transcript levels and are upregulated in a concerted manner in AD. The GFAP(+1) isoform defines a unique subset of astrocytes, with numbers increasing with AD progression. These data indicate the need for future exploration of underlying mechanisms concerning the functions of GFAPδ and GFAP(+1) isoforms in astrocytes and their possible role in AD pathology. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Roles of the troponin isoforms during indirect flight muscle development in Drosophila

    Indian Academy of Sciences (India)

    Salam Herojeet Singh; Prabodh Kumar; Nallur B. Ramachandra; Upendra Nongthomba

    2014-08-01

    Troponin proteins in cooperative interaction with tropomyosin are responsible for controlling the contraction of the striated muscles in response to changes in the intracellular calcium concentration. Contractility of the muscle is determined by the constituent protein isoforms, and the isoforms can switch over from one form to another depending on physiological demands and pathological conditions. In Drosophila, amajority of themyofibrillar proteins in the indirect flight muscles (IFMs) undergo post-transcriptional and post-translational isoform changes during pupal to adult metamorphosis to meet the high energy and mechanical demands of flight. Using a newly generated Gal4 strain (UH3-Gal4) which is expressed exclusively in the IFMs, during later stages of development, we have looked at the developmental and functional importance of each of the troponin subunits (troponin-I, troponin-T and troponin-C) and their isoforms. We show that all the troponin subunits are required for normal myofibril assembly and flight, except for the troponin-C isoform 1 (TnC1). Moreover, rescue experiments conducted with troponin-I embryonic isoform in the IFMs, where flies were rendered flightless, show developmental and functional differences of TnI isoforms and importance of maintaining the right isoform.

  2. Molecular cloning and pharmacology of functionally distinct isoforms of the human histamine H(3) receptor

    DEFF Research Database (Denmark)

    Wellendorph, Petrine; Goodman, M W; Burstein, E S

    2002-01-01

    The pharmacology of histamine H(3) receptors suggests the presence of distinct receptor isoforms or subtypes. We herein describe multiple, functionally distinct, alternatively spliced isoforms of the human H(3) receptor. Combinatorial splicing at three different sites creates at least six distinc...

  3. Roles of the troponin isoforms during indirect flight muscle development in Drosophila.

    Science.gov (United States)

    Singh, Salam Herojeet; Kumar, Prabodh; Ramachandra, Nallur B; Nongthomba, Upendra

    2014-08-01

    Troponin proteins in cooperative interaction with tropomyosin are responsible for controlling the contraction of the striated muscles in response to changes in the intracellular calcium concentration. Contractility of the muscle is determined by the constituent protein isoforms, and the isoforms can switch over from one form to another depending on physiological demands and pathological conditions. In Drosophila, amajority of themyofibrillar proteins in the indirect flight muscles (IFMs) undergo post-transcriptional and post-translational isoform changes during pupal to adult metamorphosis to meet the high energy and mechanical demands of flight. Using a newly generated Gal4 strain (UH3-Gal4) which is expressed exclusively in the IFMs, during later stages of development, we have looked at the developmental and functional importance of each of the troponin subunits (troponin-I, troponin-T and troponin-C) and their isoforms. We show that all the troponin subunits are required for normal myofibril assembly and flight, except for the troponin-C isoform 1 (TnC1). Moreover, rescue experiments conducted with troponin-I embryonic isoform in the IFMs, where flies were rendered flightless, show developmental and functional differences of TnI isoforms and importance of maintaining the right isoform.

  4. Development of isoform-specific sensors of polypeptide GalNAc-transferase activity

    DEFF Research Database (Denmark)

    Song, Lina; Bachert, Collin; Schjoldager, Katrine T

    2014-01-01

    Humans express up to 20 isoforms of GalNAc-transferase (herein T1-T20) that localize to the Golgi apparatus and initiate O-glycosylation. Regulation of this enzyme family affects a vast array of proteins transiting the secretory pathway and diseases arise upon misregulation of specific isoforms...

  5. Mitochondrial localization of the OAS1 p46 isoform associated with a common single nucleotide polymorphism

    DEFF Research Database (Denmark)

    Kjær, Karina Hansen; Pahus, Jytte; Hansen, Mariann Fagernæs

    2014-01-01

    vacuoles/lysosomes. By using recombinantly expressed OAS1 mutant proteins, we found that the OAS1 SNP rs1131454 (former rs3741981) did not affect the enzymatic OAS1 activity. Conclusions: The SNP rs10774671 determines differential expression of the OAS1 isoforms. In Daudi and HT1080 cells the p46 isoform...

  6. Comprehensive analysis of tropomyosin isoforms in skeletal muscles by top-down proteomics.

    Science.gov (United States)

    Jin, Yutong; Peng, Ying; Lin, Ziqing; Chen, Yi-Chen; Wei, Liming; Hacker, Timothy A; Larsson, Lars; Ge, Ying

    2016-04-01

    Mammalian skeletal muscles are heterogeneous in nature and are capable of performing various functions. Tropomyosin (Tpm) is a major component of the thin filament in skeletal muscles and plays an important role in controlling muscle contraction and relaxation. Tpm is known to consist of multiple isoforms resulting from different encoding genes and alternative splicing, along with post-translational modifications. However, a systematic characterization of Tpm isoforms in skeletal muscles is still lacking. Therefore, we employed top-down mass spectrometry (MS) to identify and characterize Tpm isoforms present in different skeletal muscles from multiple species, including swine, rat, and human. Our study revealed that Tpm1.1 and Tpm2.2 are the two major Tpm isoforms in swine and rat skeletal muscles, whereas Tpm1.1, Tpm2.2, and Tpm3.12 are present in human skeletal muscles. Tandem MS was utilized to identify the sequences of the major Tpm isoforms. Furthermore, quantitative analysis revealed muscle-type specific differences in the abundance of un-modified and modified Tpm isoforms in rat and human skeletal muscles. This study represents the first systematic investigation of Tpm isoforms in skeletal muscles, which not only demonstrates the capabilities of top-down MS for the comprehensive characterization of skeletal myofilament proteins but also provides the basis for further studies on these Tpm isoforms in muscle-related diseases.

  7. Translational control of C/EBPalpha and C/EBPbeta isoform expression

    NARCIS (Netherlands)

    Calkhoven, C F; Müller, C; Leutz, A

    2000-01-01

    Transcription factors derived from CCAAT/enhancer binding protein (C/EBP)alpha and C/EBPbeta genes control differentiation and proliferation in a number of cell types. Various C/EBP isoforms arise from unique C/EBPbeta and C/EBPalpha mRNAs by differential initiation of translation. These isoforms re

  8. Cloning and characterization of the neural isoforms of human dystonin

    Energy Technology Data Exchange (ETDEWEB)

    Brown, A.; Dalpe, G.; Mathieu, M.; Kothary, R. [Univ. of Montreal, Quebec (Canada)

    1995-10-10

    Dystonia musculorum (dt) is a hereditary neurodegenerative disease in mice that leads to a sensory ataxia. We have identified and cloned a gene encoded at the dt locus. The product of the dt gene, dystonin, is a neural isoform of a hemidesmosomal protein bullous pemphigoid antigen 1 (bpag1). To investigate the potential role of dystonin in human neuropathies, we have cloned the neural-specific 5{prime} exons of the human DT gene that together with the previously cloned BPAG1 sequences comprise human dystonin. The mouse and human dystonin genes demonstrate the same spectrum of alternatively spliced products, and the amino acid sequences of the neural-specific exons in the mouse and human genes are over 96% identical. 17 refs., 2 figs.

  9. Biochemical characterization of the native α-carbonic anhydrase purified from the mantle of the Mediterranean mussel, Mytilus galloprovincialis.

    Science.gov (United States)

    Perfetto, Rosa; Del Prete, Sonia; Vullo, Daniela; Sansone, Giovanni; Barone, Carmela; Rossi, Mosè; Supuran, Claudiu T; Capasso, Clemente

    2017-12-01

    A α-carbonic anhydrase (CA, EC 4.2.1.1) has been purified and characterized biochemically from the mollusk Mytilus galloprovincialis. As in most mollusks, this α-CA is involved in the biomineralization processes leading to the precipitation of calcium carbonate in the mussel shell. The new enzyme had a molecular weight of 50 kDa, which is roughly two times higher than that of a monomeric α-class enzyme. Thus, Mytilus galloprovincialis α-CA is either a dimer, or similar to the Tridacna gigas CA described earlier, may have two different CA domains in its polypeptide chain. The Mytilus galloprovincialis α-CA sequence contained the three His residues acting as zinc ligands and the gate-keeper residues present in all α-CAs (Glu106-Thr199), but had a Lys in position 64 and not a His as proton shuttling residue, being thus similar to the human isoform hCA III. This probably explains the relatively low catalytic activity of Mytilus galloprovincialis α-CA, with the following kinetic parameters for the CO2 hydration reaction: kcat = 4.1 × 10(5) s(-1) and kcat/Km of 3.6 × 10(7) M(-1) × s(-1). The enzyme activity was poorly inhibited by the sulfonamide acetazolamide, with a KI of 380 nM. This study is one of the few describing in detail the biochemical characterization of a molluskan CA and may be useful for understanding in detail the phylogeny of these enzymes, their role in biocalcification processes and their potential use in the biomimetic capture of the CO2.

  10. Identification and removal of proteins that co-purify with infectious prion protein improves the analysis of its secondary structure.

    Science.gov (United States)

    Moore, Roger A; Timmes, Andrew G; Wilmarth, Phillip A; Safronetz, David; Priola, Suzette A

    2011-10-01

    Prion diseases are neurodegenerative disorders associated with the accumulation of an abnormal isoform of the mammalian prion protein (PrP). Fourier transform infrared spectroscopy (FTIR) has previously been used to show that the conformation of aggregated, infectious PrP (PrP(Sc) ) varies between prion strains and these unique conformations may determine strain-specific disease phenotypes. However, the relative amounts of α-helix, β-sheet and other secondary structures have not always been consistent between studies, suggesting that other proteins might be confounding the analysis of PrP(Sc) secondary structure. We have used FTIR and LC-MS/MS to analyze enriched PrP(Sc) from mouse and hamster prion strains both before and after the removal of protein contaminants that commonly co-purify with PrP(Sc) . Our data show that non-PrP proteins do contribute to absorbances that have been associated with α-helical, loop, turn and β-sheet structures attributed to PrP(Sc) . The major contaminant, the α-helical protein ferritin, absorbs strongly at 1652 cm(-1) in the FTIR spectrum associated with PrP(Sc) . However, even the removal of more than 99% of the ferritin from PrP(Sc) did not completely abolish absorbance at 1652 cm(-1) . Our results show that contaminating proteins alter the FTIR spectrum attributed to PrP(Sc) and suggest that the α-helical, loop/turn and β-sheet secondary structure that remains following their removal are derived from PrP(Sc) itself.

  11. Differences in expression, actions and cocaine regulation of two isoforms for the brain transcriptional regulator NAC1.

    Science.gov (United States)

    Korutla, L; Wang, P J; Lewis, D M; Neustadter, J H; Stromberg, M F; Mackler, S A

    2002-01-01

    BTB/POZ proteins can influence the cell cycle and contribute to oncogenesis. Many family members are present in the mammalian CNS. Previous work demonstrated elevated NAC1 mRNA levels in the rat nucleus accumbens in response to cocaine. NAC1 acts like other BTB/POZ proteins that regulate transcription but is unusual because of the absence of identifiable DNA binding domains. cDNAs were isolated encoding two NAC1 isoforms differing by only 27 amino acids (the longer isoform contains 514 amino acids). The mRNAs for both isoforms were simultaneously expressed throughout the rat brain and peripheral tissues. Semi-quantitative reverse transcription-polymerase chain reaction analysis revealed that the mRNA of the longer isoform was more abundant than the mRNA of the shorter isoform. Western blot analysis demonstrated a similar unequal distribution between the isoforms in the CNS. The longer isoform was the more abundant of the two NAC1 proteins and the ratio between them differed throughout the rat brain. The shorter isoform was not detected in most of the examined peripheral tissues, suggesting differences from the CNS in post-transcriptional processing. Both isoforms repressed transcription in H293T cells using a Gal4-luciferase reporter system. However, the shorter isoform did not repress transcription as effectively as the longer isoform. Transfection of different ratios for both isoforms, in order to replicate the relative amounts observed throughout the CNS, supported an interaction between the isoforms. The net effect on transcriptional repression was determined by the ratio of the two NAC1 isoforms. Each isoform exhibited the subnuclear localization that is characteristic of many BTB/POZ proteins. A rapid and transient increase in the level of the shorter isoform occurred in the nucleus accumbens 2 h following a single i.p. cocaine injection. We conclude that the two isoforms of NAC1 may differentially affect neuronal functions, including the regulation of

  12. Studies on Purification of Methamidophos Monoclonal Antibodies and Comoarative Immunoactivity of Purified Antibodies

    Institute of Scientific and Technical Information of China (English)

    SU-QING ZHAO; YUAN-MING SUN; CHUN-YAN ZHANG; XIAO-YU HUANG; HOU-RUI ZHANG; ZHEN-YU ZHU

    2003-01-01

    Objective To purify Methamidophos (Met) monoclonal antibodies with two methods andcompare immune activity of purified antibodies. Method Caprylic acid ammonium sulphateprecipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method wereused to purify Met monoclonal antibodies, UV spectrum scanning was used to determine proteincontent and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gelelectrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linkedimmunosorbent assay (ELISA) was used to determine immune activity of purified antibodies.Results Antibody protein content and recovery rate with CAASP method were 7.62 mg/mL and8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mLand 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASPmethod. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was181.26 μg/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01μg/mL and 1.03 μg/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82μg/mL, and the linear working range and LOD were 10.91-11412.29 ug/mL and 3.42 μg/mL,respectively. Conclusion Antibodies purified by SPA method are better than those by CAASPmethod, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelledtesting paper for analyzing Met residue in vegetable and drink water.

  13. Troponin T isoform expression is modulated during Atlantic Halibut metamorphosis

    Directory of Open Access Journals (Sweden)

    Llewellyn Lynda

    2007-06-01

    Full Text Available Abstract Background Flatfish metamorphosis is a thyroid hormone (TH driven process which leads to a dramatic change from a symmetrical larva to an asymmetrical juvenile. The effect of THs on muscle and in particular muscle sarcomer protein genes is largely unexplored in fish. The change in Troponin T (TnT, a pivotal protein in the assembly of skeletal muscles sarcomeres and a modulator of calcium driven muscle contraction, during flatfish metamophosis is studied. Results In the present study five cDNAs for halibut TnT genes were cloned; three were splice variants arising from a single fast TnT (fTnT gene; a fourth encoded a novel teleost specific fTnT-like cDNA (AfTnT expressed exclusively in slow muscle and the fifth encoded the teleost specific sTnT2. THs modified the expression of halibut fTnT isoforms which changed from predominantly basic to acidic isoforms during natural and T4 induced metamorphosis. In contrast, expression of red muscle specific genes, AfTnT and sTnT2, did not change during natural metamorphosis or after T4 treatment. Prior to and after metamorphosis no change in the dorso-ventral symmetry or temporal-spatial expression pattern of TnT genes and muscle fibre organization occurred in halibut musculature. Conclusion Muscle organisation in halibut remains symmetrical even after metamorphosis suggesting TH driven changes are associated with molecular adaptations. We hypothesize that species specific differences in TnT gene expression in teleosts underlies different larval muscle developmental programs which better adapts them to the specific ecological constraints.

  14. Functional Cooperativity between ABCG4 and ABCG1 Isoforms.

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    Zoltán Hegyi

    Full Text Available ABCG4 belongs to the ABCG subfamily, the members of which are half transporters composed of a single transmembrane and a single nucleotide-binding domain. ABCG proteins have a reverse domain topology as compared to other mammalian ABC transporters, and have to form functional dimers, since the catalytic sites for ATP binding and hydrolysis, as well as the transmembrane domains are composed of distinct parts of the monomers. Here we demonstrate that ABCG4 can form homodimers, but also heterodimers with its closest relative, ABCG1. Both the full-length and the short isoforms of ABCG1 can dimerize with ABCG4, whereas the ABCG2 multidrug transporter is unable to form a heterodimer with ABCG4. We also show that contrary to that reported in some previous studies, ABCG4 is predominantly localized to the plasma membrane. While both ABCG1 and ABCG4 have been suggested to be involved in lipid transport or regulation, in accordance with our previous results regarding the long version of ABCG1, here we document that the expression of both the short isoform of ABCG1 as well as ABCG4 induce apoptosis in various cell types. This apoptotic effect, as a functional read-out, allowed us to demonstrate that the dimerization between these half transporters is not only a physical interaction but functional cooperativity. Given that ABCG4 is predominantly expressed in microglial-like cells and endothelial cells in the brain, our finding of ABCG4-induced apoptosis may implicate a new role for this protein in the clearance mechanisms within the central nervous system.

  15. Characterization of crude and purified pumpkin seed oil.

    Directory of Open Access Journals (Sweden)

    Tsaknis, John

    1997-10-01

    Full Text Available Oil from hulled pumpkin seeds (Cucurbita pepo and Cucurbita Maxima was extracted with hot petroleum ether, and then it was degummed, neutralized and bleached, consecutively Physical and chemical characteristics of crude and purified oils were determined. Density, refractive index, viscosity and peroxide value were not affected by purification, while decreases in acidity, colour, unsaponifiable, E1%1cm 232, and oxidative stability, and increases in smoke point and E1%1cm 270 were observed. Purification did not affect the fatty acid and sterol profiles. GLC analysis for the fatty acid composition of the seed oil showed that the predominant unsaturates were linoleic (42% and oleic (38%, while the major saturates were palmitic (12,7% and stearic (6%. Only α-tocopherol was detected at a level of 126 mg/kg, which reduced to 78 mg/kg after purification. The main sterols of pumpkin seed oil unsaponifiable were Δ7.22,25 -stigmastatrien-3β-ol, α-spinasterol, Δ7,25_stigmastadienol and Δ7-avenasterol, followed by stigmasterol, 24-methylcholest-7-enol and Δ7-stigmastenol, and also trace to minor amounts of cholesterol, brassicasterol, campesterol, sitostanol, Δ5-avenasterol, erythrodiol and uvaol were found.

    Aceite de semillas de calabaza descascarada (Cucurbita pepo YCucurbita maxima fue extraído con éter de petróleo caliente, y luego desgomado, neutralizado y decolorado consecutivamente. Las características físicas y químicas de aceites crudo y purificado fueron determinadas. La densidad, el índice de refracción, la viscosidad y el índice de peróxido no se afectaron por la purificación, mientras que se observó una disminución en la acidez, color, insaponificable, E1%1cm 232, y estabilidad oxidativa, y un aumento en el punto de humo y de E1%1cm270. La purificaci

  16. PURIFICATION OF WATER SOLUBLE PROTEINS (2S ALBUMINS EXTRACTED FROM PEANUT DEFATTED FLOUR AND ISOLATION OF THEIR ISOFORMS BY GEL FILTRATION AND ANION EXCHANGE CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    IMÈNE BOUALEG

    2017-06-01

    Full Text Available 2S albumins are water-soluble seed storage proteins present in dicotyledonous plants, including legumes. In peanuts, 2S albumins have been identified as major allergens. In this work, we aimed to study these water soluble allergenic proteins. They were extracted in water from peanut defatted flour (oilcake. It was quantified by Bradford method. The total and insoluble proteins content was determined by Kjeldahl method (% P = N x 6.25. The crude 2S albumins were purified using gel-filtration chromatography. Anion exchange chromatography analysis was applied to isolate their isoforms. The recorded values for total and insoluble proteins are 45.49 % and 36.65 % consecutively. A value of 9.99 % was determined for water soluble proteins content which correspond to 20 % compared to the total proteins. Analysis by Sephadex G-75 chromatography of soluble extract gave two majors peaks in which, the Mr ~ 25 kDa peak was predicted to be pure 2S albumin fraction. Using DAEA-cellulose chromatography, two peaks were appeared from pure 2S albumins, it were predicted that 2S albumin isoforms theoretically represent the peanut major allergens Ara h2 and Ara h6. These approaches are the basis for further studies may involve immunological analysis to understand the impact of these biomolecules on peanut allergenicity.

  17. Proteome from lemon fruit flavedo reveals that this tissue produces high amounts of the Cit s1 germin-like isoforms.

    Science.gov (United States)

    Pignataro, Vittorio; Canton, Cristina; Spadafora, Antonia; Mazzuca, Silvia

    2010-06-23

    A multistep procedure has been developed and applied to extract and purify proteins from lemon fruit flavedo. 2DE, LC-ESI-MS/MS, and bioinformatics were used to detect the high abundance of the germin-like glycoprotein Cit s1, a powerful allergen in humans. Peptide alignments against Citrus EST repositories gave the best scores with the C. sinensis cDNA (gi|188354270/EY710037), annotated as unknown sweet orange fruit protein; additional BLAST of peptides against NCBI databases gave high sequence identities with sequence of orange Cit s1 (gi|52782810/P84159), suggesting that the unknown sweet orange fruit protein is consistent with the Cit s1 protein. Peptides of Cit s1 were detected in 17 spots ranging from 120 to 20 kDa, pointing out that in the flavedo of lemon the Cit s1 may be expressed as several isoforms of which the 120 kDa isoform is the largest monomer and the 20 kDa is the smallest one. This finding adds information about the features of Cit s1, because it has been previously reported as a unique monomeric glycoprotein of 24 kDa.

  18. Crystallization and preliminary X-ray crystallographic analysis of latent isoform PPO4 mushroom (Agaricus bisporus) tyrosinase

    Energy Technology Data Exchange (ETDEWEB)

    Mauracher, Stephan Gerhard; Molitor, Christian [Universität Wien, Althanstrasse 14, 1090 Wien (Austria); Al-Oweini, Rami; Kortz, Ulrich [Jacobs University, PO Box 750 561, 28759 Bremen (Germany); Rompel, Annette, E-mail: annette.rompel@univie.ac.at [Universität Wien, Althanstrasse 14, 1090 Wien (Austria)

    2014-01-23

    Polyphenol oxidase 4 (PPO4) from the natural source A. bisporus was crystallized in its latent precursor form (pro-tyrosinase; Ser2–Thr565) using the 6-tungstotellurate(VI) salt Na{sub 6}[TeW{sub 6}O{sub 24}]·22H{sub 2}O as a crystallization additive. Tyrosinase exhibits catalytic activity for the ortho-hydroxylation of monophenols to diphenols as well as their subsequent oxidation to quinones. Owing to polymerization of these quinones, brown-coloured high-molecular-weight compounds called melanins are generated. The latent precursor form of polyphenol oxidase 4, one of the six tyrosinase isoforms from Agaricus bisporus, was purified to homogeneity and crystallized. The obtained crystals belonged to space group C121 (two molecules per asymmetric unit) and diffracted to 2.78 Å resolution. The protein only formed crystals under low-salt conditions using the 6-tungstotellurate(VI) salt Na{sub 6}[TeW{sub 6}O{sub 24}]·22H{sub 2}O as a co-crystallization agent.

  19. Purification and characterization of natural Ara h 8, the Bet v 1 homologous allergen from peanut, provides a novel isoform.

    Science.gov (United States)

    Riecken, Susanne; Lindner, Buko; Petersen, Arnd; Jappe, Uta; Becker, Wolf-Meinhard

    2008-04-01

    The peanut allergen Ara h 8 is an important allergen for birch pollen allergic patients because of the cross-reactivity to the homologous Bet v 1. As the existence of Ara h 8 has been shown at the cDNA level so far (AY328088) and the allergen has indirectly been detected as natural protein, it was the aim of our study to identify natural Ara h 8 in peanut extract and to develop a purification strategy. This was achieved using a unique combination of purification steps, including optimized extraction conditions, size exclusion and ion exchange chromatography and treatment of the interfering contaminants with iodoacetic acid. A characterization of the protein by microsequencing showed discrepancies to the deduced amino acid sequence of AY328088. For this reason, we cloned and expressed a new Ara h 8 isoform from cDNA (EU046325). This IgE-reactive protein corresponds to the results of microsequencing, ESI-FTICR-MS and trypsin fingerprinting analysis of the authentic and purified nAra h 8. Apart from the ultimate use of recombinant allergens for diagnostic procedures, there is also a scientific need for the natural counterpart, as it represents an excellent reference point by which to compare protein characteristics and to standardize diagnostic and therapeutic allergens.

  20. Human FAD synthase (isoform 2): a component of the machinery that delivers FAD to apo-flavoproteins.

    Science.gov (United States)

    Torchetti, Enza M; Bonomi, Francesco; Galluccio, Michele; Gianazza, Elisabetta; Giancaspero, Teresa A; Iametti, Stefania; Indiveri, Cesare; Barile, Maria

    2011-11-01

    A soluble form of human FAD synthase (isoform 2; hFADS2) was produced and purified to homogeneity as a recombinant His-tagged protein. The enzyme binds 1 mole of the FAD product very tightly, although noncovalently. Complete release of FAD from the 'as isolated' protein requires extensive denaturation. A 75 : 25 mixture of apo/holoprotein could be prepared by treatment with mild chaotropes, allowing estimatation of the contribution made by bound FAD to the protein stability and evaluatation of whether structural rearrangements may be required for FAD release. Under turnover conditions, the enzyme catalyzes FAD assembly from ATP and FMN and, at a much lower rate, the pyrophosphorolytic hydrolysis of FAD. Several mechanistic features of both reactions were investigated in detail, along with their dependence on environmental conditions (pH, temperature, dependence on metals). Our data indicate that FAD release may represent the rate-limiting step of the whole catalytic cycle and that the process leading to FAD synthesis, and delivery to client apoproteins may be tightly controlled.

  1. An experimental study on the anti-Ehrlich ascites carcinoma effect of purified toad venom extract.

    Science.gov (United States)

    Wang, Ying

    2013-01-01

    The objective of this paper was to study the anti-Ehrlich ascites carcinoma effect of purified toad venom extract and its mechanism. Mouse model of Ehrlich ascites carcinoma was established with cisplatin as the control to observe the inhibitory effect of purified toad venom extract on malignant peritoneal effusion in mice. The results showed that compared with the control group, ascites volume, number of tumour cells and tumour cell viability decreased and ascites inhibition rate reached over 50% in each treatment group, and with the increase of the dose, incidence of ascites showed a downward trend. The number of tumour cells in ascites and tumour cell viability in the purified toad venom high-dose group were lower than those of the cisplatin group. Compared with the model group, survival time was prolonged in varying degrees in the purified toad venom groups and cisplatin group. The study concluded that purified extract of toad venom has an anti-Ehrlich ascites carcinoma effect.

  2. Kinetic analysis of competition between aerosol particle removal and generation by ionization air purifiers.

    Science.gov (United States)

    Alshawa, Ahmad; Russell, Ashley R; Nizkorodov, Sergey A

    2007-04-01

    Ionization air purifiers are increasingly used to remove aerosol particles from indoor air. However, certain ionization air purifiers also emit ozone. Reactions between the emitted ozone and unsaturated volatile organic compounds (VOC) commonly found in indoor air produce additional respirable aerosol particles in the ultrafine (air purifiers under conditions of a typical residential building. This model predicts that certain widely used ionization air purifiers may actually increase the mass concentration of fine and ultrafine particulates in the presence of common unsaturated VOC, such as limonene contained in many household cleaning products. This prediction is supported by an explicit observation of ultrafine particle nucleation events caused by the addition of D-limonene to a ventilated office room equipped with a common ionization air purifier.

  3. Dystrophin Dp71 Isoforms Are Differentially Expressed in the Mouse Brain and Retina: Report of New Alternative Splicing and a Novel Nomenclature for Dp71 Isoforms.

    Science.gov (United States)

    Aragón, Jorge; González-Reyes, Mayram; Romo-Yáñez, José; Vacca, Ophélie; Aguilar-González, Guadalupe; Rendón, Alvaro; Vaillend, Cyrille; Montañez, Cecilia

    2017-01-27

    Multiple dystrophin Dp71 isoforms have been identified in rats, mice, and humans and in several cell line models. These Dp71 isoforms are produced by the alternative splicing of exons 71 to 74 and 78 and intron 77. Three main groups of Dp71 proteins are defined based on their C-terminal specificities: Dp71d, Dp71f, and Dp71e. Dp71 is highly expressed in the brain and retina; however, the specific isoforms present in these tissues have not been determined to date. In this work, we explored the expression of Dp71 isoforms in the mouse brain and retina using RT-PCR assays followed by the cloning of PCR products into the pGEM-T Easy vector, which was used to transform DH5α cells. Dp71-positive colonies were later analyzed by PCR multiplex and DNA sequencing to determine the alternative splicing. We thus demonstrated the expression of Dp71 transcripts corresponding to Dp71, Dp71a, Dp71c, Dp71b, Dp71ab, Dp71 Δ110, and novel Dp71 isoforms spliced in exon 74; 71 and 74; 71, 73 and 74; and 74 and 78, which we named Dp71d Δ74 , Dp71d Δ71,74 , Dp71d Δ71,73-74 , and Dp71f Δ74 , respectively. Additionally, we demonstrated that the Dp71d group of isoforms is highly expressed in the brain, while the Dp71f group predominates in the retina, at both the cDNA and protein levels. These findings suggest that distinct Dp71 isoforms may play different roles in the brain and retina.

  4. Serum amyloid A isoforms in serum and synovial fluid from spontaneously diseased dogs with joint diseases or other conditions

    DEFF Research Database (Denmark)

    Kjelgaard-Hansen, Mads Jens; Christensen, Michelle B.; Lee, Marcel Huisung

    2007-01-01

    with systemic inflammatory activity, and up to four major isoforms with apparent isoelectric points between 6.1 and 7.9 were identified. In synovial fluid from inflamed joints one or more highly alkaline SAA isoforms (with apparent isoelectric points above 9.3) were identified, with data suggesting local...... production of these isoforms in the canine inflamed joint....

  5. Merlin Isoforms 1 and 2 Both Act as Tumour Suppressors and Are Required for Optimal Sperm Maturation.

    Directory of Open Access Journals (Sweden)

    Ansgar Zoch

    Full Text Available The tumour suppressor Merlin, encoded by the gene NF2, is frequently mutated in the autosomal dominant disorder neurofibromatosis type II, characterised primarily by the development of schwannoma and other glial cell tumours. However, NF2 is expressed in virtually all analysed human and rodent organs, and its deletion in mice causes early embryonic lethality. Additionally, NF2 encodes for two major isoforms of Merlin of unknown functionality. Specifically, the tumour suppressor potential of isoform 2 remains controversial. In this study, we used Nf2 isoform-specific knockout mouse models to analyse the function of each isoform during development and organ homeostasis. We found that both isoforms carry full tumour suppressor functionality and can completely compensate the loss of the other isoform during development and in most adult organs. Surprisingly, we discovered that spermatogenesis is strictly dependent on the presence of both isoforms. While the testis primarily expresses isoform 1, we noticed an enrichment of isoform 2 in spermatogonial stem cells. Deletion of either isoform was found to cause decreased sperm quality as observed by maturation defects and head/midpiece abnormalities. These defects led to impaired sperm functionality as assessed by decreased sperm capacitation. Thus, we describe spermatogenesis as a new Nf2-dependent process. Additionally, we provide for the first time in vivo evidence for equal tumour suppressor potentials of Merlin isoform 1 and isoform 2.

  6. Role of nuclear progesterone receptor isoforms in uterine pathophysiology.

    Science.gov (United States)

    Patel, Bansari; Elguero, Sonia; Thakore, Suruchi; Dahoud, Wissam; Bedaiwy, Mohamed; Mesiano, Sam

    2015-01-01

    Progesterone is a key hormonal regulator of the female reproductive system. It plays a major role to prepare the uterus for implantation and in the establishment and maintenance of pregnancy. Actions of progesterone on the uterine tissues (endometrium, myometrium and cervix) are mediated by the combined effects of two progesterone receptor (PR) isoforms, designated PR-A and PR-B. Both receptors function primarily as ligand-activated transcription factors. Progesterone action on the uterine tissues is qualitatively and quantitatively determined by the relative levels and transcriptional activities of PR-A and PR-B. The transcriptional activity of the PR isoforms is affected by specific transcriptional coregulators and by PR post-translational modifications that affect gene promoter targeting. In this context, appropriate temporal and cell-specific expression and function of PR-A and PR-B are critical for normal uterine function. Relevant studies describing the role of PRs in uterine physiology and pathology (endometriosis, uterine leiomyoma, endometrial cancer, cervical cancer and recurrent pregnancy loss) were comprehensively searched using PubMed, Cochrane Library, Web of Science, and Google Scholar and critically reviewed. Progesterone, acting through PR-A and PR-B, regulates the development and function of the endometrium and induces changes in cells essential for implantation and the establishment and maintenance of pregnancy. During pregnancy, progesterone via the PRs promotes myometrial relaxation and cervical closure. Withdrawal of PR-mediated progesterone signaling triggers menstruation and parturition. PR-mediated progesterone signaling is anti-mitogenic in endometrial epithelial cells, and as such, mitigates the tropic effects of estrogen on eutopic normal endometrium, and on ectopic implants in endometriosis. Similarly, ligand-activated PRs function as tumor suppressors in endometrial cancer cells through inhibition of key cellular signaling pathways

  7. Dynamic expression and localization of c-MET isoforms in the developing rat pancreas.

    Science.gov (United States)

    Wu, Yulong; Cheng, Mei; Shi, Zhen; Feng, Zhenqing; Guan, Xiaohong

    2014-01-01

    Pancreata from Sprague Dawley rats of different developmental stages were studied to determine the expression and cellular localization of different c-MET isoforms in the developing rat pancreas. Pancreatic mRNA and protein expression levels of c-MET at different developmental stages from embryo to adult were detected by reverse transcription-polymerase chain reaction and by western blotting. To identify the cellular localization of c-MET protein in the developing rat pancreas, double immunofluorescent staining was performed using antibodies for cell type-specific markers and for c-MET. The expression of two isoforms of c-MET (190 kDa and 170 kDa) coincided with the development of the pancreas. The 190 kDa isoform of c-MET is expressed during embryonic stages, and its expression is replaced by the expression of the 170 kDa isoform as the pancreas develops. Only the 170 kDa isoform is expressed in the adult rat pancreas. Throughout all stages of pancreatic development, c-MET is expressed by vimentin-positive cells. In contrast, c-MET staining was stronger in rat pancreata from newborn to adult stages and overlapped with insulin-positive beta-cells. The dynamic expression and localization of different c-MET isoforms in the rat pancreas during different developmental stages indicates that distinct c-MET isoform might be involved in different aspects of pancreatic development.

  8. Distinct and shared transcriptomes are regulated by microphthalmia-associated transcription factor isoforms in mast cells.

    Science.gov (United States)

    Shahlaee, Amir H; Brandal, Stephanie; Lee, Youl-Nam; Jie, Chunfa; Takemoto, Clifford M

    2007-01-01

    The Microphthalmia-associated transcription factor (Mitf) is an essential basic helix-loop-helix leucine zipper transcription factor for mast cell development. Mice deficient in Mitf harbor a severe mast cell deficiency, and Mitf-mutant mast cells cultured ex vivo display a number of functional defects. Therefore, an understanding of the genetic program regulated by Mitf may provide important insights into mast cell differentiation. Multiple, distinct isoforms of Mitf have been identified in a variety of cell types; we found that Mitf-a, Mitf-e, and Mitf-mc were the major isoforms expressed in mast cells. To determine the physiologic function of Mitf in mast cells, we restored expression of these isoforms in primary mast cells from Mitf(-/-) mice. We found that these isoforms restored granular morphology and integrin-mediated migration. By microarray analysis, proteases, signaling molecules, cell surface receptor, and transporters comprised the largest groups of genes up-regulated by all isoforms. Furthermore, we found that isoforms also regulated distinct genes sets, suggesting separable biological activities. This work defines the transcriptome regulated by Mitf in mast cells and supports its role as master regulator of mast cell differentiation. Expression of multiple isoforms of this transcription factor may provide for redundancy of biological activities while also allowing diversity of function.

  9. VEGF-A isoforms program differential VEGFR2 signal transduction, trafficking and proteolysis

    Science.gov (United States)

    Fearnley, Gareth W.; Smith, Gina A.; Abdul-Zani, Izma; Yuldasheva, Nadira; Mughal, Nadeem A.; Homer-Vanniasinkam, Shervanthi; Kearney, Mark T.; Zachary, Ian C.; Tomlinson, Darren C.; Harrison, Michael A.; Wheatcroft, Stephen B.; Ponnambalam, Sreenivasan

    2016-01-01

    ABSTRACT Vascular endothelial growth factor A (VEGF-A) binding to the receptor tyrosine kinase VEGFR2 triggers multiple signal transduction pathways, which regulate endothelial cell responses that control vascular development. Multiple isoforms of VEGF-A can elicit differential signal transduction and endothelial responses. However, it is unclear how such cellular responses are controlled by isoform-specific VEGF-A–VEGFR2 complexes. Increasingly, there is the realization that the membrane trafficking of receptor–ligand complexes influences signal transduction and protein turnover. By building on these concepts, our study shows for the first time that three different VEGF-A isoforms (VEGF-A165, VEGF-A121 and VEGF-A145) promote distinct patterns of VEGFR2 endocytosis for delivery into early endosomes. This differential VEGFR2 endocytosis and trafficking is linked to VEGF-A isoform-specific signal transduction events. Disruption of clathrin-dependent endocytosis blocked VEGF-A isoform-specific VEGFR2 activation, signal transduction and caused substantial depletion in membrane-bound VEGFR1 and VEGFR2 levels. Furthermore, such VEGF-A isoforms promoted differential patterns of VEGFR2 ubiquitylation, proteolysis and terminal degradation. Our study now provides novel insights into how different VEGF-A isoforms can bind the same receptor tyrosine kinase and elicit diverse cellular outcomes. PMID:27044325

  10. VEGF-A isoforms program differential VEGFR2 signal transduction, trafficking and proteolysis

    Directory of Open Access Journals (Sweden)

    Gareth W. Fearnley

    2016-05-01

    Full Text Available Vascular endothelial growth factor A (VEGF-A binding to the receptor tyrosine kinase VEGFR2 triggers multiple signal transduction pathways, which regulate endothelial cell responses that control vascular development. Multiple isoforms of VEGF-A can elicit differential signal transduction and endothelial responses. However, it is unclear how such cellular responses are controlled by isoform-specific VEGF-A–VEGFR2 complexes. Increasingly, there is the realization that the membrane trafficking of receptor–ligand complexes influences signal transduction and protein turnover. By building on these concepts, our study shows for the first time that three different VEGF-A isoforms (VEGF-A165, VEGF-A121 and VEGF-A145 promote distinct patterns of VEGFR2 endocytosis for delivery into early endosomes. This differential VEGFR2 endocytosis and trafficking is linked to VEGF-A isoform-specific signal transduction events. Disruption of clathrin-dependent endocytosis blocked VEGF-A isoform-specific VEGFR2 activation, signal transduction and caused substantial depletion in membrane-bound VEGFR1 and VEGFR2 levels. Furthermore, such VEGF-A isoforms promoted differential patterns of VEGFR2 ubiquitylation, proteolysis and terminal degradation. Our study now provides novel insights into how different VEGF-A isoforms can bind the same receptor tyrosine kinase and elicit diverse cellular outcomes.

  11. Estrogen and progesterone receptor isoforms expression in the stomach of Mongolian gerbils

    Institute of Scientific and Technical Information of China (English)

    Milena Saqui-Salces; Teresa Neri-Gómez; Armando Gamboa-Dominguez; Guillermo Ruiz-Palacios; Ignacio Camacho-Arroyo

    2008-01-01

    AIM:We studied the estrogen receptor (ER) and progesterone receptor (PR) isoforms expression in gastric antrum and corpus of female gerbils and their regulation by estradiol (E2) and progesterone (P4).METHODS: Ovariectomized adult female gerbils were subcutaneously treated with E2,and E2 + P4.Uteri and stomachs were removed,the latter were cut along the greater curvature,and antrum and corpus were excised.Proteins were immunoblotted using antibodies that recognize ER-alpha,ER-beta,and PR-A and PR-B receptor isoforms.Tissues from rats treated in the same way were used as controls.RESULTS: Specific bands were detected for ER-alpha (68 KDa),and PR isoforms (85 and 120 KDa for PR-A and PR-B isoforrns,respectively) in uteri,gastric antrum and corpus.We could not detect ER-beta isoform.PR isoforms were not regulated by E2 or P4 in uterus and gastric tissues of gerbils.ER-alpha isoform content was significantly down-regulated by E2 in the corpus,but not affected by hormones in uterus and gastric antrum.CONCLUSION: The presence of ER-alpha and PR isoforms in gerbils stomach suggests that E2 and P4 actions in this organ are in part mediated by their nuclear receptors.

  12. Identification and evolutionary analysis of tissue-specific isoforms of mitochondrial complex I subunit NDUFV3.

    Science.gov (United States)

    Guerrero-Castillo, Sergio; Cabrera-Orefice, Alfredo; Huynen, Martijn A; Arnold, Susanne

    2017-03-01

    Mitochondrial complex I is the largest respiratory chain complex. Despite the enormous progress made studying its structure and function in recent years, potential regulatory roles of its accessory subunits remained largely unresolved. Complex I gene NDUFV3, which occurs in metazoa, contains an extra exon that is only present in vertebrates and thereby evolutionary even younger than the rest of the gene. Alternative splicing of this extra exon gives rise to a short NDUFV3-S and a long NDUFV3-L protein isoform. Complexome profiling revealed that the two NDUFV3 isoforms are constituents of the multi-subunit complex I. Further mass spectrometric analyses of complex I from different murine and bovine tissues showed a tissue-specific expression pattern of NDUFV3-S and NDUFV3-L. Hence, NDUFV3-S was identified as the only isoform in heart and skeletal muscle, whereas in liver, brain, and lung NDUFV3-L was expressed as the dominant isoform, together with NDUFV3-S present in all tissues analyzed. Thus, we identified NDUFV3 as the first out of 30 accessory subunits of complex I present in vertebrate- and tissue-specific isoforms. Interestingly, the tissue-specific expression pattern of NDUFV3-S and NDUFV3-L isoforms was paralleled by changes in kinetic parameters, especially the substrate affinity of complex I. This may indicate a regulatory role of the NDUFV3 isoforms in different vertebrate tissues.

  13. Revisiting the identification of canonical splice isoforms through integration of functional genomics and proteomics evidence.

    Science.gov (United States)

    Li, Hong-Dong; Menon, Rajasree; Omenn, Gilbert S; Guan, Yuanfang

    2014-12-01

    Canonical isoforms in different databases have been defined as the most prevalent, most conserved, most expressed, longest, or the one with the clearest description of domains or posttranslational modifications. In this article, we revisit these definitions of canonical isoforms based on functional genomics and proteomics evidence, focusing on mouse data. We report a novel functional relationship network-based approach for identifying the highest connected isoforms (HCIs). We show that 46% of these HCIs are not the longest transcripts. In addition, this approach revealed many genes that have more than one highly connected isoforms. Averaged across 175 RNA-seq datasets covering diverse tissues and conditions, 65% of the HCIs show higher expression levels than nonhighest connected isoforms at the transcript level. At the protein level, these HCIs highly overlap with the expressed splice variants, based on proteomic data from eight different normal tissues. These results suggest that a more confident definition of canonical isoforms can be made through integration of multiple lines of evidence, including HCIs defined by biological processes and pathways, expression prevalence at the transcript level, and relative or absolute abundance at the protein level. This integrative proteogenomics approach can successfully identify principal isoforms that are responsible for the canonical functions of genes. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Channel properties of the splicing isoforms of the olfactory calcium-activated chloride channel Anoctamin 2.

    Science.gov (United States)

    Ponissery Saidu, Samsudeen; Stephan, Aaron B; Talaga, Anna K; Zhao, Haiqing; Reisert, Johannes

    2013-06-01

    Anoctamin (ANO)2 (or TMEM16B) forms a cell membrane Ca(2+)-activated Cl(-) channel that is present in cilia of olfactory receptor neurons, vomeronasal microvilli, and photoreceptor synaptic terminals. Alternative splicing of Ano2 transcripts generates multiple variants with the olfactory variants skipping exon 14 and having alternative splicing of exon 4. In the present study, 5' rapid amplification of cDNA ends analysis was conducted to characterize the 5' end of olfactory Ano2 transcripts, which showed that the most abundant Ano2 transcripts in the olfactory epithelium contain a novel starting exon that encodes a translation initiation site, whereas transcripts of the publically available sequence variant, which has an alternative and longer 5' end, were present in lower abundance. With two alternative starting exons and alternative splicing of exon 4, four olfactory ANO2 isoforms are thus possible. Patch-clamp experiments in transfected HEK293T cells expressing these isoforms showed that N-terminal sequences affect Ca(2+) sensitivity and that the exon 4-encoded sequence is required to form functional channels. Coexpression of the two predominant isoforms, one with and one without the exon 4 sequence, as well as coexpression of the two rarer isoforms showed alterations in channel properties, indicating that different isoforms interact with each other. Furthermore, channel properties observed from the coexpression of the predominant isoforms better recapitulated the native channel properties, suggesting that the native channel may be composed of two or more splicing isoforms acting as subunits that together shape the channel properties.

  15. Androgen receptor isoforms in human prostatic cancer tissue and LNCaP cell line

    Institute of Scientific and Technical Information of China (English)

    Shu-Jie XIA; Xiao-Da TANG; Qing-Zheng MA

    2001-01-01

    Aim: To investigate the androgen receptor (AR) isoform expressions in human prostatic cancer tissue and LNCaP cell line. Methods: With high resolution isoelectric focusing (IEF) method we demonstrated the different expressions of AR isoforms in human prostatic cancer tissues and LNCaP cell line. Results: Data were obtained from three prostatic cancer specimens and the LNCaP cell line. Three types of AR isoforms were detected with pI values at 6.5,6.0, and 5.3. For the 3 prostatic cancer specimens, 1 sample showed all the three types of AR isoforms, the second specimen expressed at 6.5 and 6.0, and the third failed to show any type of isoforms. The LNCaP cell line expressed all the three AR isoforms. Binding of 3H-dihydrotestosterone (3H-DHT) to these three isoforms was inhibited by the addition ofl00-fold excess of DHT or testosterone, while not by progesterone, oestradiol and diethylstilboestrol. Conclusion: The expression of AR isofonns is different in different prostate cancer tissues, which may be related to the difference in the effect of anti-androgen therapy in different patients.

  16. Differential subcellular distribution of four phospholipase C isoforms and secretion of GPI-PLC activity.

    Science.gov (United States)

    Staudt, Emanuel; Ramasamy, Pathmanaban; Plattner, Helmut; Simon, Martin

    2016-12-01

    Phospholipase C (PLC) is an important enzyme of signal transduction pathways by generation of second messengers from membrane lipids. PLCs are also indicated to cleave glycosylphosphatidylinositol (GPI)-anchors of surface proteins thus releasing these into the environment. However, it remains unknown whether this enzymatic activity on the surface is due to distinct PLC isoforms in higher eukaryotes. Ciliates have, in contrast to other unicellular eukaryotes, multiple PLC isoforms as mammals do. Thus, Paramecium represents a perfect model to study subcellular distribution and potential surface activity of PLC isoforms. We have identified distinct subcellular localizations of four PLC isoforms indicating functional specialization. The association with different calcium release channels (CRCs) argues for distinct subcellular functions. They may serve as PI-PLCs in microdomains for local second messenger responses rather than free floating IP3. In addition, all isoforms can be found on the cell surface and they are found together with GPI-cleaved surface proteins in salt/ethanol washes of cells. We can moreover show them in medium supernatants of living cells where they have access to GPI-anchored surface proteins. Among the isoforms we cannot assign GPI-PLC activity to specific PLC isoforms; rather each PLC is potentially responsible for the release of GPI-anchored proteins from the surface. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Inulin isoforms differ by repeated additions of one crystal unit cell.

    Science.gov (United States)

    Cooper, Peter D; Barclay, Thomas G; Ginic-Markovic, Milena; Gerson, Andrea R; Petrovsky, Nikolai

    2014-03-15

    Inulin isoforms, especially delta inulin, are important biologically as immune activators and clinically as vaccine adjuvants. In exploring action mechanisms, we previously found regular increments in thermal properties of the seven-member inulin isoform series that suggested regular additions of some energetic structural unit. Because the previous isolates carried additional longer chains that masked defining ranges, these were contrasted with new isoform isolates comprising only inulin chain lengths defining that isoform. The new series began with 19 fructose units per chain (alpha-1 inulin), increasing regularly by 6 fructose units per isoform. Thus the 'energetic unit' equates to 6 fructose residues per chain. All isoforms showed indistinguishable X-ray diffraction patterns that were also identical with known inulin crystals. We conclude that an 'energetic unit' equates to one helix turn of 6 fructose units per chain as found in one unit cell of the inulin crystal. Each isoform chain comprised progressively more helix turns plus one additional fructose and glucose residues per chain.

  18. Detection of VEGF-A(xxx)b isoforms in human tissues.

    Science.gov (United States)

    Bates, David O; Mavrou, Athina; Qiu, Yan; Carter, James G; Hamdollah-Zadeh, Maryam; Barratt, Shaney; Gammons, Melissa V; Millar, Ann B; Salmon, Andrew H J; Oltean, Sebastian; Harper, Steven J

    2013-01-01

    Vascular Endothelial Growth Factor-A (VEGF-A) can be generated as multiple isoforms by alternative splicing. Two families of isoforms have been described in humans, pro-angiogenic isoforms typified by VEGF-A165a, and anti-angiogenic isoforms typified by VEGF-A165b. The practical determination of expression levels of alternative isoforms of the same gene may be complicated by experimental protocols that favour one isoform over another, and the use of specific positive and negative controls is essential for the interpretation of findings on expression of the isoforms. Here we address some of the difficulties in experimental design when investigating alternative splicing of VEGF isoforms, and discuss the use of appropriate control paradigms. We demonstrate why use of specific control experiments can prevent assumptions that VEGF-A165b is not present, when in fact it is. We reiterate, and confirm previously published experimental design protocols that demonstrate the importance of using positive controls. These include using known target sequences to show that the experimental conditions are suitable for PCR amplification of VEGF-A165b mRNA for both q-PCR and RT-PCR and to ensure that mispriming does not occur. We also provide evidence that demonstrates that detection of VEGF-A165b protein in mice needs to be tightly controlled to prevent detection of mouse IgG by a secondary antibody. We also show that human VEGF165b protein can be immunoprecipitated from cultured human cells and that immunoprecipitating VEGF-A results in protein that is detected by VEGF-A165b antibody. These findings support the conclusion that more information on the biology of VEGF-A165b isoforms is required, and confirm the importance of the experimental design in such investigations, including the use of specific positive and negative controls.

  19. Distinctive effects of rat fibroblast growth factor-2 isoforms on PC12 and Schwann cells.

    Science.gov (United States)

    Müller-Ostermeyer, F; Claus, P; Grothe, C

    2001-01-01

    Fibroblast growth factor-2 (FGF-2) is an important modulator of cell growth and differentiation and stimulates cell survival of various cells including neurons. Rat FGF-2 occurs in three isoforms, a low molecular weight 18 kD and two high molecular weight forms (21, 23 kD), representing alternative translation products from a single mRNA. The 18 kD isoform shows mainly cytoplasmatic localization, whereas the 21/23 kD FGF-2 are localized in the nucleus. In addition, the FGF-2 isoforms are differentially regulated in the sensory ganglia and peripheral nerve following nerve injury and in the adrenal medulla during post-natal development and after hormonal stimuli. The distinct intracellular distribution and differential regulation of the different FGF-2 isoforms indicate that they have unique biological roles, however, little is known about the biological effects of the high molecular weight FGF-2 isoforms. Immortalized Schwann cells and PC12 cells, which stably overexpress the different FGF-2 isoforms, showed that the different endogenous-overexpressed FGF-2 isoforms lead to dramatic modifications in cell proliferation and survival, when tested in serum-free and serum-containing medium. In contrast, application of recombinant FGF-2 isoforms on normal PC12 and immortalized Schwann cells results in similar biological effects on the proliferation and survival of the cells. Furthermore, we investigated the potential regulatory effects of endogenous-overexpressed and exogenous-applied FGF-2 isoforms on the mRNA level of the FGF-2 receptors and, additionally, on the tyrosin hydroxylase mRNA expression in PC12 cells.

  20. Non-muscle Myosin II Isoforms Co-assemble in Living Cells

    Science.gov (United States)

    Beach, Jordan R.; Shao, Lin; Remmert, Kirsten; Li, Dong; Betzig, Eric; Hammer, John A.

    2014-01-01

    SUMMARY Non-muscle myosin II (NM II) powers myriad developmental and cellular processes, including embryogenesis, cell migration, and cytokinesis [1]. To exert its functions, monomers of NM II assemble into bipolar filaments that produce a contractile force on the actin cytoskeleton. Mammalian cells express up to three isoforms of NM II (NM IIA, IIB and IIC), each of which possesses distinct biophysical properties and supports unique, as well as redundant, cellular functions [2-8]. Despite previous efforts [9-13], it remains unclear if NM II isoforms assemble in living cells to produce mixed (heterotypic) bipolar filaments, or if filaments consist entirely of a single isoform (homotypic). We addressed this question using fluorescently-tagged versions of NM IIA, IIB and IIC, isoform-specific immunostaining of the endogenous proteins, and two-color total internal reflection fluorescence structured-illumination microscopy, or TIRF-SIM, to visualize individual myosin II bipolar filaments inside cells. We show that NM II isoforms co-assemble into heterotypic filaments in a variety of settings, including various types of stress fibers, individual filaments throughout the cell, and the contractile ring. We also show that the differential distribution of NM IIA and NM IIB typically seen in confocal micrographs of well-polarized cells is reflected in the composition of individual bipolar filaments. Interestingly, this differential distribution is less pronounced in freshly-spread cells, arguing for the existence of sorting mechanism acting over time. Together, our work argues that individual NM II isoforms are potentially performing both isoform-specific and isoform-redundant functions while co-assembled with other NM II isoforms. PMID:24814144