Multiplexed Molecular Assays for Rapid Rule-Out of Foot-and-Mouth Disease (open access)

Multiplexed Molecular Assays for Rapid Rule-Out of Foot-and-Mouth Disease

A nucleic acid-based multiplexed assay was developed that combines detection of foot-and-mouth disease virus (FMDV) with rule-out assays for two other foreign animal diseases and four domestic animal diseases that cause vesicular or ulcerative lesions indistinguishable from FMDV infection in cattle, sheep and swine. The FMDV 'look-alike' diagnostic assay panel contains five PCR and twelve reverse transcriptase PCR (RT-PCR) signatures for a total of seventeen simultaneous PCR amplifications for seven diseases plus incorporating four internal assay controls. It was developed and optimized to amplify both DNA and RNA viruses simultaneously in a single tube and employs Luminex{trademark} liquid array technology. Assay development including selection of appropriate controls, a comparison of signature performance in single and multiplex testing against target nucleic acids, as well of limits of detection for each of the individual signatures is presented. While this assay is a prototype and by no means a comprehensive test for FMDV 'look-alike' viruses, an assay of this type is envisioned to have benefit to a laboratory network in routine surveillance and possibly for post-outbreak proof of freedom from foot-and-mouth disease.
Date: June 26, 2007
Creator: Lenhoff, R; Naraghi-Arani, P; Thissen, J; Olivas, J; Carillo, C; Chinn, C et al.
Object Type: Article
System: The UNT Digital Library
High-Efficiency Solar Cells for Large-Scale Electricity Generation (open access)

High-Efficiency Solar Cells for Large-Scale Electricity Generation

One strategy for helping the solar industry to grow faster is to use very high efficiency cells under concentrating optics. By using lenses or mirrors to concentrate the light, very small solar cells can be used, reducing the amount of semiconductor material and allowing use of higher efficiency cells, which are now >40% efficient.
Date: September 26, 2008
Creator: Kurtz, S.; Olson, J.; Geisz, J.; Friedman, D.; McMahon, W.; Ptak, A. et al.
Object Type: Article
System: The UNT Digital Library
EUV mask reflectivity measurements with micron-scale spatial resolution (open access)

EUV mask reflectivity measurements with micron-scale spatial resolution

The effort to produce defect-free mask blanks for EUV lithography relies on increasing the detection sensitivity of advanced mask inspection tools, operating at several wavelengths. We describe the unique measurement capabilities of a prototype actinic (EUV wavelength) microscope that is capable of detecting small defects and reflectivity changes that occur on the scale of microns to nanometers. Types of defects: (a) Buried Substrate Defects: particles & pits (causes amplitude and/or phase variations); (b) Surface Contamination (reduces reflectivity and (possibly) contrast); (c) Damage from Inspection and Use (reduces the reflectivity of the multilayer coating). This paper presents an overview of several topics where scanning actinic inspection makes a unique contribution to EUVL research. We describe the role of actinic scanning inspection in four cases: defect repair studies; observations of laser damage; after scanning electron microscopy; and native and programmed defects.
Date: May 26, 2008
Creator: Goldberg, Kenneth A.; Rekawa, S. B.; Kemp, C. D.; Barty, A.; Anderson, E. H.; Kearney, Patrick et al.
Object Type: Article
System: The UNT Digital Library
AMMONIA CONCENTRATION IN SALTSTONE HEADSPACE SUMMARY REPORT (open access)

AMMONIA CONCENTRATION IN SALTSTONE HEADSPACE SUMMARY REPORT

The Saltstone Facility Documented Safety Analysis (DSA) is under revision to accommodate changes in the Composite Lower Flammability Limit (CLFL) from the introduction of Isopar into Tank 50. Saltstone samples were prepared with an 'MCU' type salt solution spiked with ammonia. The ammonia released from the saltstone was captured and analyzed. The ammonia concentration found in the headspace of samples maintained at 95 C and 1 atm was, to 95% confidence, less than or equal to 3.9 mg/L. Tank 50 is fed by several influent streams. The salt solution from Tank 50 is pumped to the salt feed tank (SFT) in the Saltstone Production Facility (SPF). The premix materials cement, slag and fly ash are blended together prior to transfer to the grout mixer. The premix is fed to the grout mixer in the SPF and the salt solution is incorporated into the premix in the grout mixer, yielding saltstone slurry. The saltstone slurry drops into a hopper and then is pumped to the vault. The Saltstone Facility Documented Safety Analysis (DSA) is under revision to accommodate changes in the Composite Lower Flammability Limit (CLFL) from the introduction of Isopar{reg_sign} L into Tank 50. Waste Solidification-Engineering requested that the Savannah …
Date: September 26, 2008
Creator: Zamecnik, J & Alex Cozzi, A
Object Type: Report
System: The UNT Digital Library
Hanford Site Environmental Surveillance Data Report for Calendar Year 2006 (open access)

Hanford Site Environmental Surveillance Data Report for Calendar Year 2006

This document includes tables of analytical data for Hanford Site environmental surveillance samples collected in 2006.
Date: September 26, 2007
Creator: Bisping, Lynn E.
Object Type: Report
System: The UNT Digital Library
PDV Probe Alignment Technique (open access)

PDV Probe Alignment Technique

This alignment technique was developed while performing heterodyne velocimetry measurements at LLNL. There are a few minor items needed, such as a white card with aperture in center, visible alignment laser, IR back reflection meter, and a microscope to view the bridge surface. The work was performed on KCP flyers that were 6 and 8 mils wide. The probes used were Oz Optics manufactured with focal distances of 42mm and 26mm. Both probes provide a spot size of approximately 80?m at 1550nm. The 42mm probes were specified to provide an internal back reflection of -35 to -40dB, and the probe back reflections were measured to be -37dB and -33dB. The 26mm probes were specified as -30dB and both measured -30.5dB. The probe is initially aligned normal to the flyer/bridge surface. This provides a very high return signal, up to -2dB, due to the bridge reflectivity. A white card with a hole in the center as an aperture can be used to check the reflected beam position relative to the probe and launch beam, and the alignment laser spot centered on the bridge, see Figure 1 and Figure 2. The IR back reflection meter is used to measure the dB return …
Date: October 26, 2007
Creator: Whitworth, T L; May, C M & Strand, O T
Object Type: Report
System: The UNT Digital Library
Colliding with a crunching bubble (open access)

Colliding with a crunching bubble

In the context of eternal inflation we discuss the fate of Lambda = 0 bubbles when they collide with Lambda< 0 crunching bubbles. When the Lambda = 0 bubble is supersymmetric, it is not completely destroyed by collisions. If the domain wall separating the bubbles has higher tension than the BPS bound, it is expelled from the Lambda = 0 bubble and does not alter its long time behavior. If the domain wall saturates the BPS bound, then it stays inside the Lambda = 0 bubble and removes a finite fraction of future infinity. In this case, the crunch singularity is hidden behind the horizon of a stable hyperbolic black hole.
Date: March 26, 2007
Creator: Freivogel, Ben; Freivogel, Ben; Horowitz, Gary T. & Shenker, Stephen
Object Type: Article
System: The UNT Digital Library
B365 High Efficiency Particulate Air Filter Emergency Replacement An Example of Team Work (open access)

B365 High Efficiency Particulate Air Filter Emergency Replacement An Example of Team Work

None
Date: September 26, 2006
Creator: Coble, T; Little, C & Johnson, R
Object Type: Article
System: The UNT Digital Library
Beam Collimation Studies for the ILC Positron Source (open access)

Beam Collimation Studies for the ILC Positron Source

Results of the collimation studies for the ILC positron source beam line are presented. The calculations of primary positron beam loss are done using the ELEGANT code. The secondary positron and electron beam loss, the synchrotron radiation along the beam line and the bremsstrahlung radiation in the collimators are simulated using the STRUCT code. The first part of the collimation system, located right after the positron source target (0.125 GeV), is used for protection of the RF Linac sections from heating and radiation. The second part of the system is used for final collimation before the beam injection into the Damping Ring at 5 GeV. The calculated power loss in the collimation region is within 100 W/m, with the loss in the collimators of 0.2-5 kW. The beam transfer efficiency from the target to the Damping Ring is 13.5%.
Date: June 26, 2008
Creator: Drozhdin, A.; Nosochkov, Y. & Zhou, F.
Object Type: Article
System: The UNT Digital Library
Green (2(omega)) Laser Beam propagation in high-temperature Hohlraum Plasmas (open access)

Green (2(omega)) Laser Beam propagation in high-temperature Hohlraum Plasmas

We demonstrate propagation and small backscatter losses of a frequency-doubled (2{omega}) laser beam interacting with inertial confinement fusion hohlraum plasmas. The electron temperature of 3.3 keV, approximately a factor of two higher than achieved in previous experiments with open geometry targets, approaches plasma conditions of high-fusion yield hohlraums. In this new temperature regime, we measure 2{omega} laser beam transmission approaching 80% with simultaneous backscattering losses of less than 10%. These findings suggests that good laser coupling into fusion hohlraums using 2{omega} light is possible.
Date: October 26, 2007
Creator: Niemann, C; Berger, R; Divol, L; Froula, D H; Jones, O S; Kirkwood, R K et al.
Object Type: Article
System: The UNT Digital Library
Measurement of the Hadronic Mass Spectrum in B to Xulnu Decaysand Determination of the b-Quark Mass at the BaBar Experiment (open access)

Measurement of the Hadronic Mass Spectrum in B to Xulnu Decaysand Determination of the b-Quark Mass at the BaBar Experiment

I present preliminary results of the measurement of the hadronic mass spectrum and its first three spectral moments in inclusive charmless semileptonic B-meson decays. The truncated hadronic mass moments are used for the first determination of the b-quark mass and the nonperturbative parameters {mu}{sub {pi}}{sup 2} and {rho}{sub D}{sup 3} in this B-meson decay channel. The study is based on 383 x 10{sup 6} B{bar B} decays collected with the BABAR experiment at the PEP-II e{sup +}e{sup -} storage rings, located at the Stanford Linear Accelerator Center. The first, second central, and third central hadronic mass moment with a cut on the hadronic mass m{sub X}{sup 2} < 6.4GeV{sup 2} and the lepton momentum p* > 1 GeV are measured to be: M{sub 1} = (1.96 {+-} 0.34{sub stat} {+-} 0.53{sub syst}) GeV{sup 2}; U{sub 2} = (1.92 {+-} 0.59{sub stat} {+-} 0.87{sub syst}) GeV{sup 4}; and U{sub 3} = (1.79 {+-} 0.62{sub stat} {+-} 0.78{sub syst}) GeV{sup 6}; with correlation coefficients {rho}{sub 12} = 0.99, {rho}{sub 23} = 0.94, and {rho}{sub 13} = 0.88, respectively. Using Heavy Quark Effective Theory-based predictions in the kinetic scheme we extract: m{sub b} = (4.60 {+-} 0.13{sub stat} {+-} 0.19{sub syst} {+-} 0.10{sub …
Date: June 26, 2008
Creator: Tackmann, Kerstin & /UC, Berkeley /SLAC
Object Type: Thesis or Dissertation
System: The UNT Digital Library
Acclerator R&D for a Linear Collider (open access)

Acclerator R&D for a Linear Collider

The goal of this project was to perform simulations of beam transport in linear colliders, with an emphasis on emittance dilution, spin polarization transport, and development and testing of beam based tuning algorithms. Our simulations are based on an existing object-oriented particle-tracking library, Bmad. To facilitate the efficient development of simulations, an accelerator design and analysis program based on Bmad has been developed called Tao (Tool for Accelerator Optics). The three beam-based alignment algorithms, Dispersion Free Steering, Ballistic Alignment (BA), and the Kubo Method have been implemented in Tao. We have studied the effects of magnet misalignments, BPM resolution, beam jitter, stray fields, BPM and steering magnet failure and the effects of various cavity shape wakefields. A parametric study has been conducted in the presence of the above types of errors for all three alignment algorithms. We find that BPM resolution has only modest impact on the effectiveness of beam based alignment. The DFS correction algorithm was found to be very robust in situations where there were BPM and/or steering magnet failures. The wakefields in the main linac are very weak and cause negligible emittance growth. Spin tracking was extended to study all accelerator components between the damping ring and …
Date: November 26, 2008
Creator: Rubin, D.L.; Dugan, G.; Gibbons, L.; Palmer, M.; Patterson, R.; Sagan, D. et al.
Object Type: Report
System: The UNT Digital Library
Full-Process Computer Model of Magnetron Sputter, Part I: Test Existing State-of-Art Components (open access)

Full-Process Computer Model of Magnetron Sputter, Part I: Test Existing State-of-Art Components

This work is part of a larger project to develop a modeling capability for magnetron sputter deposition. The process is divided into four steps: plasma transport, target sputter, neutral gas and sputtered atom transport, and film growth, shown schematically in Fig. 1. Each of these is simulated separately in this Part 1 of the project, which is jointly funded between CMLS and Engineering. The Engineering portion is the plasma modeling, in step 1. The plasma modeling was performed using the Object-Oriented Particle-In-Cell code (OOPIC) from UC Berkeley [1]. Figure 2 shows the electron density in the simulated region, using magnetic field strength input from experiments by Bohlmark [2], where a scale of 1% is used. Figures 3 and 4 depict the magnetic field components that were generated using two-dimensional linear interpolation of Bohlmark's experimental data. The goal of the overall modeling tool is to understand, and later predict, relationships between parameters of film deposition we can change (such as gas pressure, gun voltage, and target-substrate distance) and key properties of the results (such as film stress, density, and stoichiometry.) The simulation must use existing codes, either open-source or low-cost, not develop new codes. In part 1 (FY07) we identified and …
Date: September 26, 2007
Creator: Walton, C. C.; Gilmer, G. H.; Wemhoff, A. P. & Zepeda-Ruiz, L. A.
Object Type: Report
System: The UNT Digital Library
Thoughts on Verification of Nuclear Disarmament (open access)

Thoughts on Verification of Nuclear Disarmament

It is my pleasure to be here to day to participate in this Conference. My thanks to the organizers for preparing such an interesting agenda on a very difficult topic. My effort in preparing my presentation was performed under the auspices of the U.S. Department of Energy by University of California, Lawrence Livermore National Laboratory under Contract W-7405-Eng-48. And as many of you know Lawrence Livermore National Laboratory is now, as of Oct 1st, under contract to the Lawrence Livermore National Security LLC. There has been a long history of how to view verification of arms control agreements. The basis for verification during the days of SALT was that verification would be based on each country's national technical means. For treaties dealing with strategic missiles this worked well as the individual items subject to verification were of such a size that they were visible by the National Technical Means available at the time. And it was felt that the counting of missiles and launchers could be verified by our National Technical Means. For nuclear testing treaties the use of seismic measurements developed into a capability that was reasonably robust for all but the smallest of nuclear tests. However, once we …
Date: September 26, 2007
Creator: Dunlop, W H
Object Type: Article
System: The UNT Digital Library
Physics Design Requirements for the National Spherical Torus Experiment Liquid Lithium Divertor (open access)

Physics Design Requirements for the National Spherical Torus Experiment Liquid Lithium Divertor

Recent NSTX high power divertor experiments have shown significant and recurring benefits of solid lithium coatings on PFC's to the performance of divertor plasmas in both L- and H- mode confinement regimes heated by high-power neutral beams. The next step in this work is installation of a liquid lithium divertor (LLD) to achieve density control for inductionless current drive capability (e.g., about a 15-25% ne decrease from present highest non-inductionless fraction discharges which often evolve toward the density limit, ne/nGW~1), to enable ne scan capability (x2) in the H-mode, to test the ability to operate at significantly lower density for future ST-CTF reactor designs (e.g., ne/nGW = 0.25), and eventually to investigate high heat-flux power handling (10 MW/m2) with longpulse discharges (>1.5s). The first step (LLD-1) physics design encompasses the desired plasma requirements, the experimental capabilities and conditions, power handling, radial location, pumping capability, operating temperature, lithium filling, MHD forces, and diagnostics for control and characterization.
Date: September 26, 2008
Creator: Kugel, W.; Bell, M.; Berzak,L.; Brooks, A.; Ellis, R.; Gerhardt, S. et al.
Object Type: Article
System: The UNT Digital Library
Evaluating EUV mask pattern imaging with two EUV microscopes (open access)

Evaluating EUV mask pattern imaging with two EUV microscopes

Aerial image measurement plays a key role in the development of patterned reticles for each generation of lithography. Studying the field transmitted (reflected) from EUV masks provides detailed information about potential disruptions caused by mask defects, and the performance of defect repair strategies, without the complications of photoresist imaging. Furthermore, by measuring the continuously varying intensity distribution instead of a thresholded, binary resist image, aerial image measurement can be used as feedback to improve mask and lithography system modeling methods. Interest in EUV, at-wavelength, aerial image measurement lead to the creation of several research tools worldwide. These tools are used in advanced mask development work, and in the evaluation of the need for commercial at-wavelength inspection tools. They describe performance measurements of two such tools, inspecting the same EUV mask in a series of benchmarking tests that includes brightfield and darkfield patterns. One tool is the SEMATECH Berkeley Actinic Inspection Tool (AIT) operating on a bending magnet beamline at Lawrence Berkeley National Laboratory's Advanced Light Source. The AIT features an EUV Fresnel zoneplate microscope that emulates the numerical aperture of a 0.25-NA stepper, and projects the aerial image directly onto a CCD camera, with 700x magnification. The second tool is …
Date: February 26, 2008
Creator: Goldberg, Kenneth A.; Takase, Kei; Naulleau, Patrick P.; Han, Hakseung; Barty, Anton; Kinoshita, Hiroo et al.
Object Type: Article
System: The UNT Digital Library
An Octadentate Luminescent Eu(III) 1,2-HOPO Chelate with PotentAqueous Stability (open access)

An Octadentate Luminescent Eu(III) 1,2-HOPO Chelate with PotentAqueous Stability

None
Date: February 26, 2007
Creator: Moore, Evan G.; Jocher, Christoph J.; Xu, Jide; Werner, Eric J. & Raymond,Kenneth N.
Object Type: Article
System: The UNT Digital Library
Visualization of Excitonic Structure in the Fenna-Matthews-OlsonPhotosynthetic Complex by Polarization-Dependent Two-DimensionalElectronic Spectroscopy (open access)

Visualization of Excitonic Structure in the Fenna-Matthews-OlsonPhotosynthetic Complex by Polarization-Dependent Two-DimensionalElectronic Spectroscopy

Photosynthetic light-harvesting proceeds by the collection and highly efficient transfer of energy through a network of pigment-protein complexes. Inter-chromophore electronic couplings and interactions between pigments and the surrounding protein determine energy levels of excitonic states and dictate the mechanism of energy flow. The excitonic structure (orientation of excitonic transition dipoles) of pigment-protein complexes is generally deduced indirectly from x-ray crystallography in combination with predictions of transition energies and couplings in the chromophore site basis. Here, we demonstrate that coarse-grained excitonic structural information in the form of projection angles between transition dipole moments can be obtained from polarization-dependent two-dimensional electronic spectroscopy of an isotropic sample, particularly when the nonrephasing or free polarization decay signal rather than the photon echo signal is considered. The method provides an experimental link between atomic and electronic structure and accesses dynamical information with femtosecond time resolution. In an investigation of the Fenna-Matthews-Olson complex from green sulfur bacteria, energy transfer connecting two particular exciton states in the protein is isolated as being the primary contributor to a cross peak in the nonrephasing 2D spectrum at 400 fs under a specific sequence of polarized excitation pulses. The results suggest the possibility of designing experiments using combinations of tailored …
Date: May 26, 2008
Creator: Department of Chemistry, The University of Chicago; Department of Biology, Department of Chemistry, Washington University; Fleming, Graham; Read, Elizabeth L.; Schlau-Cohen, Gabriela S.; Engel, Gregory S. et al.
Object Type: Article
System: The UNT Digital Library
Poland Becoming a Member of the Global Nuclear Energy Partnership, Vol. 1. (open access)

Poland Becoming a Member of the Global Nuclear Energy Partnership, Vol. 1.

Within a constrained carbon environment, the risks of future natural gas supply, and the need to move to market-based electricity prices, the study team found: (1) the deployment of new nuclear energy in Poland itself is very competitive in the next decade or two; (2) if such generation could be made available to Poland prior to deployment of its own nuclear generation facilities, Poland would benefit from partnering with its Baltic neighbors to import electricity derived from new nuclear generation facilities sited in Lithuania; and (3) Poland appears to be a good candidate for a partnership in the Global Nuclear Energy Partnership (GNEP) as an emerging nuclear energy country.
Date: March 26, 2007
Creator: Koritarov, V. K.; Conzelmann, G.; Cirillo, R. R. & Goldberg, S. M.
Object Type: Report
System: The UNT Digital Library
Remaining Sites Verification Package for the 1607-F3 Sanitary Sewer System, Waste Site Reclassification Form 2006-047 (open access)

Remaining Sites Verification Package for the 1607-F3 Sanitary Sewer System, Waste Site Reclassification Form 2006-047

The 1607-F3 waste site is the former location of the sanitary sewer system that supported the 182-F Pump Station, the 183-F Water Treatment Plant, and the 151-F Substation. The sanitary sewer system included a septic tank, drain field, and associated pipeline, all in use between 1944 and 1965. In accordance with this evaluation, the verification sampling results support a reclassification of this site to Interim Closed Out. The results of verification sampling demonstrated that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results also showed that residual contaminant concentrations are protective of groundwater and the Columbia River.
Date: April 26, 2007
Creator: Dittmer, L. M.
Object Type: Report
System: The UNT Digital Library
Detailed Comparison of Blast Effects in Air and Vacuum (open access)

Detailed Comparison of Blast Effects in Air and Vacuum

Although blast mitigation is most often achieved with solid shielding, ambient gas pressure can also affect the coupling of shock waves to solid targets. In this work the role of air as an energy transfer medium was examined experimentally by subjecting identical large-area rectangular witness plates to short-range blast effects in air and vacuum ({approx}50 mtorr) at 25 C. The expanding reactant front of 3 kg C4 charges was observed by fast camera to be cylindrically symmetric in both air and vacuum. The horizontal component of the reactant cloud velocity (perpendicular to the witness plates) was constant in both cases, with values of 3.0 and 5.9 km/s for air and vacuum, respectively. As a result of the blast, witness plates were plastically deformed into a shallow dish geometry, with local maxima 30 and 20 mm deep for air and vacuum, respectively. The average plate deflection from the air blast was 11 mm, {approx}10% deeper than the average vacuum plate deflection. Shock pressure estimates were made with a simple impedance-matching model, and indicate peak values in the 30-50 MPa range are consistent with the reactant cloud density and velocity. However, more detailed analysis is necessary to definitely establish the mechanisms by …
Date: July 26, 2007
Creator: Tringe, J W; Molitoris, J D; Garza, R G; Andreski, H G; Batteux, J D; Lauderbach, L M et al.
Object Type: Article
System: The UNT Digital Library
Local Structure of La1-xSrxCoO3 determined from EXAFS and neutron PDF studies (open access)

Local Structure of La1-xSrxCoO3 determined from EXAFS and neutron PDF studies

The combined local structure techniques, extended x-ray absorption fine structure (EXAFS) and neutron pair distribution function analysis, have been used for temperatures 4<= T<= 330 K to rule out a large Jahn-Teller (JT) distortion of the Co-O bond in La1?xSrxCoO3 for a significant fraction of Co sites (x<= 0.35), indicating few, if any, JT-active, singly occupied eg Co sites exist.
Date: January 26, 2009
Creator: Sundaram, N.; Jiang, Y.; Anderson, I. E.; Belanger, D. P.; Booth, C. H.; Bridges, F. et al.
Object Type: Article
System: The UNT Digital Library
UNITED STATES DEPARTMENT OF ENERGY OFFICE OF ENVIRONMENTAL MANAGEMENT INTERNATIONAL PROGRAM ANNUAL REPORT FOR 2007 (open access)

UNITED STATES DEPARTMENT OF ENERGY OFFICE OF ENVIRONMENTAL MANAGEMENT INTERNATIONAL PROGRAM ANNUAL REPORT FOR 2007

The DOE-EM Office of Engineering and Technology is responsible for implementing EM's international cooperative program. The Office of Engineering and Technology's international efforts are aimed at supporting EM's mission of risk reduction and accelerated cleanup of the environmental legacy of the nation's nuclear weapons program and government-sponsored nuclear energy research. To do this, EM pursues collaborations with government organizations, educational institutions, and private industry to identify and develop technologies that can address the site cleanup needs of DOE. The Office of Engineering and Technology has developed a Technology Roadmap and a Multi-year Program Plan to identify technology needs and identify areas for focused research and development to support DOE-EM's environmental cleanup and waste management objectives. The international cooperative program is an important element of the technology development roadmap, leveraging of world-wide expertise in the advancement and deployment of remediation and treatment technologies. Introductory briefings aimed at furthering familiarity with the DOE-EM mission, and the vital role that technology development plays within it, were presented at two international meetings. The Office of Engineering and Technology currently works with the Khlopin Radium Institute (KRI) and SIA Radon Institute in Russia, the International Radioecology Laboratory (IRL) in Ukraine and the Nuclear Engineering and …
Date: August 26, 2008
Creator: Marra, J
Object Type: Report
System: The UNT Digital Library
Proteomic Analysis of Calcium- and Phosphorylation-dependentCalmodulin Complexes in Mammalian Cells (open access)

Proteomic Analysis of Calcium- and Phosphorylation-dependentCalmodulin Complexes in Mammalian Cells

Protein conformational changes due to cofactor binding (e.g. metal ions, heme) and/or posttranslational modifications (e.g. phosphorylation) modulate dynamic protein complexes. Calmodulin (CaM) plays an essential role in regulating calcium (Ca{sup 2+}) signaling and homeostasis. No systematic approach on the identification of phosphorylation-dependent Ca{sup 2+}/CaM binding proteins has been published. Herein, we report a proteome-wide study of phosphorylation-dependent CaM binding proteins from mammalian cells. This method, termed 'Dynamic Phosphoprotein Complex Trapping', 'DPPC Trapping' for short, utilizes a combination of in vivo and in vitro assays. The basic strategy is to drastically shift the equilibrium towards endogenous phosphorylation of Ser, Thr, and Tyr at the global scale by inhibiting corresponding phosphatases in vivo. The phosphorylation-dependent calmodulin-binding proteins are then trapped in vitro in a Ca{sup 2+}-dependent manner by CaM-Sepharose chromatography. Finally, the isolated calmodulin-binding proteins are separated by SDS-PAGE and identified by LC/MS/MS. In parallel, the phosphorylation-dependent binding is visualized by silver staining and/or Western blotting. Using this method, we selectively identified over 120 CaM-associated proteins including many previously uncharacterized. We verified ubiquitin-protein ligase EDD1, inositol 1, 4, 5-triphosphate receptor type 1 (IP{sub 3}R1), and ATP-dependent RNA helicase DEAD box protein 3 (DDX3), as phosphorylation-dependent CaM binding proteins. To demonstrate the utilities …
Date: May 26, 2006
Creator: Jang, Deok-Jin & Wang, Daojing
Object Type: Article
System: The UNT Digital Library