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Sample records for nuclear hot cell

  1. 48 CFR 952.225-70 - Subcontracting for nuclear hot cell services.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... hot cell services. 952.225-70 Section 952.225-70 Federal Acquisition Regulations System DEPARTMENT OF....225-70 Subcontracting for nuclear hot cell services. As prescribed in 925.7004, insert the following clause in solicitations and contracts: Subcontracting for Nuclear Hot Cell Services (MAR 1993)...

  2. 48 CFR 952.225-70 - Subcontracting for nuclear hot cell services.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... from nuclear reactors, which are performed in specialized facilities located away from commercial... 48 Federal Acquisition Regulations System 5 2012-10-01 2012-10-01 false Subcontracting for nuclear....225-70 Subcontracting for nuclear hot cell services. As prescribed in 925.7004, insert the...

  3. 48 CFR 952.225-70 - Subcontracting for nuclear hot cell services.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... from nuclear reactors, which are performed in specialized facilities located away from commercial... 48 Federal Acquisition Regulations System 5 2014-10-01 2014-10-01 false Subcontracting for nuclear....225-70 Subcontracting for nuclear hot cell services. As prescribed in 925.7004, insert the...

  4. 48 CFR 952.225-70 - Subcontracting for nuclear hot cell services.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... from nuclear reactors, which are performed in specialized facilities located away from commercial... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Subcontracting for nuclear....225-70 Subcontracting for nuclear hot cell services. As prescribed in 925.7004, insert the...

  5. 48 CFR 952.225-70 - Subcontracting for nuclear hot cell services. >

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... from nuclear reactors, which are performed in specialized facilities located away from commercial... 48 Federal Acquisition Regulations System 5 2011-10-01 2011-10-01 false Subcontracting for nuclear... Clauses 952.225-70 Subcontracting for nuclear hot cell services.> As prescribed in 925.7004, insert...

  6. Reliable Wireless Data Acquisition and Control Techniques within Nuclear Hot Cell Facilities

    SciTech Connect

    Kurtz, J.L.; Tulenko, J.

    2000-09-20

    On this NEER project the University of Florida has investigated and applied advanced communications techniques to address data acquisition and control problems within the Fuel Conditioning Facility (FCF) of Argonne National Laboratory-West (ANL-W) in Idaho Falls. The goals of this project have been to investigate and apply wireless communications techniques to solve the problem of communicating with and controlling equipment and systems within a nuclear hot cell facility with its attendant high radiation levels. Different wireless techniques, including radio frequency, infrared and power line communications were reviewed. For each technique, the challenges of radiation-hardened implementation were addressed. In addition, it has been a project goal to achieve the highest level of system reliability to ensure safe nuclear operations. Achievement of these goals would allow the eventual elimination of through-the-wall, hardwired cabling that is currently employed in the hot cell, along wit h all of the attendant problems that limit measurement mobility and flexibility.

  7. Nuclear Materials Characterization in the Materials and Fuels Complex Analytical Hot Cells

    SciTech Connect

    Michael Rodriquez

    2009-03-01

    As energy prices skyrocket and interest in alternative, clean energy sources builds, interest in nuclear energy has increased. This increased interest in nuclear energy has been termed the “Nuclear Renaissance”. The performance of nuclear fuels, fuels and reactor materials and waste products are becoming a more important issue as the potential for designing new nuclear reactors is more immediate. The Idaho National Laboratory (INL) Materials and Fuels Complex (MFC) Analytical Laboratory Hot Cells (ALHC) are rising to the challenge of characterizing new reactor materials, byproducts and performance. The ALHC is a facility located near Idaho Falls, Idaho at the INL Site. It was built in 1958 as part of the former Argonne National Laboratory West Complex to support the operation of the second Experimental Breeder Reactor (EBR-II). It is part of a larger analytical laboratory structure that includes wet chemistry, instrumentation and radiochemistry laboratories. The purpose of the ALHC is to perform analytical chemistry work on highly radioactive materials. The primary work in the ALHC has traditionally been dissolution of nuclear materials so that less radioactive subsamples (aliquots) could be transferred to other sections of the laboratory for analysis. Over the last 50 years though, the capabilities within the ALHC have also become independent of other laboratory sections in a number of ways. While dissolution, digestion and subdividing samples are still a vitally important role, the ALHC has stand alone capabilities in the area of immersion density, gamma scanning and combustion gas analysis. Recent use of the ALHC for immersion density shows that extremely fine and delicate operations can be performed with the master-slave manipulators by qualified operators. Twenty milligram samples were tested for immersion density to determine the expansion of uranium dioxide after irradiation in a nuclear reactor. The data collected confirmed modeling analysis with very

  8. Hot cell examination table

    DOEpatents

    Gaal, Peter S.; Ebejer, Lino P.; Kareis, James H.; Schlegel, Gary L.

    1991-01-01

    A table for use in a hot cell or similar controlled environment for use in examining specimens. The table has a movable table top that can be moved relative to a table frame. A shaft is fixedly mounted to the frame for axial rotation. A shaft traveler having a plurality of tilted rollers biased against the shaft is connected to the table top such that rotation of the shaft causes the shaft traveler to roll along the shaft. An electromagnetic drive is connected to the shaft and the frame for controllably rotating the shaft.

  9. Seismic evaluation of a hot cell structure

    SciTech Connect

    Srinivasan, M.G.; Kot, C.A.

    1995-07-01

    The evaluation of the structural capacity of and the seismic demand on an existing hot cell structure in a nuclear facility is described. An ANSYS finite-element model of the cell was constructed, treating the walls as plates and the floor and ceiling as a system of discrete beams. A modal analysis showed that the fundamental frequencies of the cell walls lie far above the earthquake frequency range. An equivalent static analysis of the structure was performed. Based on the analysis it was demonstrated that the hot cell structure, would readily withstand the evaluation basis earthquake.

  10. DECOMMISSIONING OF HOT CELL FACILITIES AT THE BATTELLE COLUMBUS LABORATORIES

    SciTech Connect

    Weaver, Patrick; Henderson, Glenn; Erickson, Peter; Garber, David

    2003-02-27

    Battelle Columbus Laboratories (BCL), located in Columbus, Ohio, must complete decontamination and decommissioning activities for nuclear research buildings and grounds at its West Jefferson Facilities by 2006, as mandated by Congress. This effort includes decommissioning several hot cells located in the Hot Cell Laboratory (Building JN-1). JN-1 was originally constructed in 1955, and a hot cell/high bay addition was built in the mid 1970s. For over 30 years, BCL used these hot cell facilities to conduct research for the nuclear power industry and several government agencies, including the U.S. Navy, U.S. Army, U.S. Air Force, and the U.S. Department of Energy. As a result of this research, the JN-1 hot cells became highly contaminated with mixed fission and activation products, as well as fuel residues. In 1998, the Battelle Columbus Laboratories Decommissioning Project (BCLDP) began efforts to decommission JN-1 with the goal of remediating the site to levels of residual contamination allowing future use without radiological restrictions. This goal requires that each hot cell be decommissioned to a state where it can be safely demolished and transported to an off-site disposal facility. To achieve this, the BCLDP uses a four-step process for decommissioning each hot cell: (1) Source Term Removal; (2) Initial (i.e., remote) Decontamination; (3) Utility Removal; and (4) Final (i.e., manual) Decontamination/Stabilization. To date, this process has been successfully utilized on 13 hot cells within JN-1, with one hot cell remaining to be decommissioned. This paper will provide a case study of the hot cell decommissioning being conducted by the BCLDP. Discussed will be the methods used to achieve the goals of each of the hot cell decommissioning stages and the lessons learned that could be applied at other sites where hot cells need to be decommissioned.

  11. Hot Cell Facility (HCF) Safety Analysis Report

    SciTech Connect

    MITCHELL,GERRY W.; LONGLEY,SUSAN W.; PHILBIN,JEFFREY S.; MAHN,JEFFREY A.; BERRY,DONALD T.; SCHWERS,NORMAN F.; VANDERBEEK,THOMAS E.; NAEGELI,ROBERT E.

    2000-11-01

    This Safety Analysis Report (SAR) is prepared in compliance with the requirements of DOE Order 5480.23, Nuclear Safety Analysis Reports, and has been written to the format and content guide of DOE-STD-3009-94 Preparation Guide for U. S. Department of Energy Nonreactor Nuclear Safety Analysis Reports. The Hot Cell Facility is a Hazard Category 2 nonreactor nuclear facility, and is operated by Sandia National Laboratories for the Department of Energy. This SAR provides a description of the HCF and its operations, an assessment of the hazards and potential accidents which may occur in the facility. The potential consequences and likelihood of these accidents are analyzed and described. Using the process and criteria described in DOE-STD-3009-94, safety-related structures, systems and components are identified, and the important safety functions of each SSC are described. Additionally, information which describes the safety management programs at SNL are described in ancillary chapters of the SAR.

  12. Modified Nuclear Lifetime in Hot Dense Plasmas

    NASA Astrophysics Data System (ADS)

    Gosselin, Gilbert; Meot, V.; Morel, P.

    In plasmas, the electronic environment in the immediate vicinity of the nucleus is modified, and thus, the plasma conditions influence key processes driving the lifetime of a nuclear level. A correct lifetime prediction requires every de-excitation process to be evaluated jointly with its corresponding excitation process. For heavy nuclei, the nuclear lifetime of discrete levels is often strongly dependent on internal conversion which involves bound electrons. In plasma, many of these electrons are no longer in a bound state and the internal conversion rate can be significantly reduced. Its coupling with its inverse process, Nuclear Excitation by Electron Capture (NEEC), can lead to greatly increased nuclear lifetimes. In some cases, an atomic transition can be coupled with a nuclear transition in a process called Nuclear Excitation by Electron Transition (NEET) if their energies are closely matched. This can accelerate the de-excitation of the excited nuclear level, and reduce its lifetime. We developed a model able to deal with these processes in plasma under thermodynamic equilibrium. It evaluates internal conversion, NEEC and NEET rates in plasma. Depending on the particular situation, we use either an average atom description or a Multi Configuration Dirac Fock (MCDF) approach to describe the electronic environment of the atom. Large variations of several excited nuclear level lifetimes have been predicted. A complete description of the nuclear lifetime must also include some other nuclear levels through which indirect nuclear excitation or de-excitation may occur. This particular situation may provide a fast method to populate or depopulate nuclear isomers.

  13. Hot Cell Window Shielding Analysis Using MCNP

    SciTech Connect

    Chad L. Pope; Wade W. Scates; J. Todd Taylor

    2009-05-01

    The Idaho National Laboratory Materials and Fuels Complex nuclear facilities are undergoing a documented safety analysis upgrade. In conjunction with the upgrade effort, shielding analysis of the Fuel Conditioning Facility (FCF) hot cell windows has been conducted. This paper describes the shielding analysis methodology. Each 4-ft thick window uses nine glass slabs, an oil film between the slabs, numerous steel plates, and packed lead wool. Operations in the hot cell center on used nuclear fuel (UNF) processing. Prior to the shielding analysis, shield testing with a gamma ray source was conducted, and the windows were found to be very effective gamma shields. Despite these results, because the glass contained significant amounts of lead and little neutron absorbing material, some doubt lingered regarding the effectiveness of the windows in neutron shielding situations, such as during an accidental criticality. MCNP was selected as an analysis tool because it could model complicated geometry, and it could track gamma and neutron radiation. A bounding criticality source was developed based on the composition of the UNF. Additionally, a bounding gamma source was developed based on the fission product content of the UNF. Modeling the windows required field inspections and detailed examination of drawings and material specifications. Consistent with the shield testing results, MCNP results demonstrated that the shielding was very effective with respect to gamma radiation, and in addition, the analysis demonstrated that the shielding was also very effective during an accidental criticality.

  14. Stress analysis for wall structure in mobile hot cell design

    SciTech Connect

    Bahrin, Muhammad Hannan Rahman, Anwar Abdul Hamzah, Mohd Arif Mamat, Mohd Rizal; Azman, Azraf; Hasan, Hasni

    2016-01-22

    Malaysian Nuclear Agency is developing a Mobile Hot Cell (MHC) in order to handle and manage Spent High Activity Radioactive Sources (SHARS) such as teletherapy heads and irradiators. At present, there are only two units of MHC in the world, in South Africa and China. Malaysian Mobile Hot cell is developed by Malaysian Nuclear Agency with the assistance of IAEA expert, based on the design of South Africa and China, but with improved features. Stress analysis has been performed on the design in order to fulfil the safety requirement in operation of MHC. This paper discusses the loading analysis effect from the sand to the MHC wall structure.

  15. Hot magnetized nuclear matter: Thermodynamic and saturation properties

    NASA Astrophysics Data System (ADS)

    Rezaei, Z.; Bordbar, G. H.

    2017-03-01

    We have used a realistic nuclear potential, AV_{18}, and a many-body technique, the lowest-order constraint variational (LOCV) approach, to calculate the properties of hot magnetized nuclear matter. By investigating the free energy, spin polarization parameter, and symmetry energy, we have studied the temperature and magnetic field dependence of the saturation properties of magnetized nuclear matter. In addition, we have calculated the equation of state of magnetized nuclear matter at different temperatures and magnetic fields. It was found that the flashing temperature of nuclear matter decreases by increasing the magnetic field. In addition, we have studied the effect of the magnetic field on liquid gas phase transition of nuclear matter. The liquid gas coexistence curves, the order parameter of the liquid gas phase transition, and the properties of critical point at different magnetic fields have been calculated.

  16. Pressurized water nuclear reactor system with hot leg vortex mitigator

    DOEpatents

    Lau, Louis K. S.

    1990-01-01

    A pressurized water nuclear reactor system includes a vortex mitigator in the form of a cylindrical conduit between the hot leg conduit and a first section of residual heat removal conduit, which conduit leads to a pump and a second section of residual heat removal conduit leading back to the reactor pressure vessel. The cylindrical conduit is of such a size that where the hot leg has an inner diameter D.sub.1, the first section has an inner diameter D.sub.2, and the cylindrical conduit or step nozzle has a length L and an inner diameter of D.sub.3 ; D.sub.3 /D.sub.1 is at least 0.55, D.sub.2 is at least 1.9, and L/D.sub.3 is at least 1.44, whereby cavitation of the pump by a vortex formed in the hot leg is prevented.

  17. Equation of state for {beta}-stable hot nuclear matter

    SciTech Connect

    Moustakidis, Ch. C.; Panos, C. P.

    2009-04-15

    We provide an equation of state for hot nuclear matter in {beta} equilibrium by applying a momentum-dependent effective interaction. We focus on the study of the equation of state of high-density and high-temperature nuclear matter, containing leptons (electrons and muons) under the chemical equilibrium condition in which neutrinos have left the system. The conditions of charge neutrality and equilibrium under the {beta}-decay process lead first to the evaluation of proton and lepton fractions and then to the evaluation of internal energy, free energy, and pressure, and in total to the equation of state of hot nuclear matter. Thermal effects on the properties and equation of state of nuclear matter are assessed and analyzed in the framework of the proposed effective interaction model. Special attention is given to the study of the contribution of the components of {beta}-stable nuclear matter to the entropy per particle, a quantity of great interest in the study of structure and collapse of supernova.

  18. Long Duration Hot Hydrogen Exposure of Nuclear Thermal Rocket Materials

    NASA Technical Reports Server (NTRS)

    Litchford, Ron J.; Foote, John P.; Hickman, Robert; Dobson, Chris; Clifton, Scooter

    2007-01-01

    An arc-heater driven hyper-thermal convective environments simulator was recently developed and commissioned for long duration hot hydrogen exposure of nuclear thermal rocket materials. This newly established non-nuclear testing capability uses a high-power, multi-gas, wall-stabilized constricted arc-heater to .produce high-temperature pressurized hydrogen flows representative of nuclear reactor core environments, excepting radiation effects, and is intended to serve as a low cost test facility for the purpose of investigating and characterizing candidate fuel/structural materials and improving associated processing/fabrication techniques. Design and engineering development efforts are fully summarized, and facility operating characteristics are reported as determined from a series of baseline performance mapping runs and long duration capability demonstration tests.

  19. Thermodynamic instabilities in hot and dense nuclear matter

    NASA Astrophysics Data System (ADS)

    Lavagno, A.

    2016-11-01

    We study the presence of thermodynamic instabilities in a hot and dense nuclear medium where a nuclear phase transition can take place. Similarly to the low density nuclear liquid-gas phase transition, we show that such a phase transition is characterized by pure hadronic matter with both mechanical instability (fluctuations on the baryon density) that by chemical-diffusive instability (fluctuations on the strangeness concentration). The analysis is performed by requiring the global conservation of baryon number and zero net strangeness in the framework of an effective relativistic mean field theory with the inclusion of the Δ(1232)-isobars, hyperons and the lightest pseudoscalar and vector meson degrees of freedom. It turns out that in this situation hadronic phases with different values of strangeness content may coexist, altering significantly meson-antimeson ratios.

  20. JINA Workshop Nuclear Physics in Hot Dense Dynamic Plasmas

    SciTech Connect

    Kritcher, A L; Cerjan, C; Landen, O; Libby, S; Chen, M; Wilson, B; Knauer, J; Mcnabb, D; Caggiano, J; Bleauel, D; Weideking, M; Kozhuharov, C; Brandau, C; Stoehlker, T; Meot, V; Gosselin, G; Morel, P; Schneider, D; Bernstein, L A

    2011-03-07

    Measuring NEET and NEEC is relevant for probing stellar cross-sections and testing atomic models in hot plasmas. Using NEEC and NEET we can excite nuclear levels in laboratory plasmas: (1) NIF: Measure effect of excited nuclear levels on (n,{gamma}) cross-sections, 60% and never been measured; (2) Omega, Test cross-sections for creating these excited levels via NEEC and NEET. Will allow us to test models that estimate resonance overlap of atomic states with the nucleus: (1) Average Atom model (AA) (CEA&LLNL), single average wave-function potential; (2) Super Transition Array (STA) model (LLNL), More realistic individual configuration potentials NEET experimental data is scarce and not in a plasma environment, NEEC has not yet been observed.

  1. Nuclear Reactions in Hot Astrophysical Plasmas with T > 1010 K

    NASA Astrophysics Data System (ADS)

    Kafexhiu, Ervin; Aharonian, Felix; Vila, Gabriela S.

    The importance of nuclear reactions in low-density astrophysical plasmas with ion temperatures T ≥1010 K has been recognized for more than thirty years. However, the lack of comprehensive data banks of relevant nuclear reactions and the limited computational power have not previously allowed detailed theoretical studies. Recent developments in these areas make it timely to conduct comprehensive studies on the nuclear properties of very hot plasmas formed around compact relativistic objects such as black holes and neutron stars. Such studies are of great interest in the context of scientific programs of future low-energy cosmic γ-ray spectrometry. In this work, using the publicly available code TALYS, we have built a large nuclear network relevant for temperatures exceeding 1010 K. We have studied the evolution of the chemical composition and accompanying prompt gamma-ray emission of such high-temperature plasmas. We present the results on the abundances of light elements D, T, 3He, 4He, 6Li, 7Li, 9Be, 10B, 11B, and briefly discuss their implications on the astrophysical abundances of these elements.

  2. Hot nuclear matter in the static fluctuation approximation

    SciTech Connect

    Ghulam, N. M.; Ghassib, H. B.; Al-Sugheir, M. K.

    2007-06-15

    In this work, the bulk and thermodynamic properties of nonrelativistic hot nuclear matter--the mean internal energy per unit volume, the saturation density and the corresponding internal energy per nucleon, the pressure, the entropy per unit volume, the heat capacity per unit volume, and the chemical potential--are studied within the static fluctuation approximation (SFA). The basic input is the well-known Reid68 and Reid93 soft-core potentials, with special emphasis on three channels that have different spin and isospin--namely, the {sup 1}S{sub 0}-channel as well as the {sup 3}S{sub 1}-{sup 3}D{sub 1} and {sup 3}P{sub 2}-{sup 3}F{sub 2} coupled channels. Finally, a full-fledged calculation is presented using the Reid93 potential for all channels J{<=}2. Wherever possible, comparisons are made with previous calculations. It is concluded that SFA is valid for hot nuclear matter over a wide range of temperatures ({<=}50 MeV)

  3. A&M. Hot cell annex (TAN633) interior under construction. Hot cells ...

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

    A&M. Hot cell annex (TAN-633) interior under construction. Hot cells and their doors are along concrete wall. Note side wall of pumice block. Photographer: Jack L. Anderson. Date: October 28, 1957. INEEL negative no. 57-5335 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  4. HOT CELL BUILDING, TRA632. SHIELDING DOOR TO HOT CELL IS ...

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

    HOT CELL BUILDING, TRA-632. SHIELDING DOOR TO HOT CELL IS IN OPEN POSITION. DOOR SLIDES SHUT WITH HELP OF MANUALLY OPERATED CHAIN. STAIRWAY TO MEZZANINE IN VIEW AT LEFT. CAMERA FACES NORTHWEST. INL NEGATIVE NO. 9000. Unknown Photographer, 10/28/1953 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  5. HOT CELL BUILDING, TRA632. HOT CELL AWAITS INSTALLATION OF SHIELDED ...

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

    HOT CELL BUILDING, TRA-632. HOT CELL AWAITS INSTALLATION OF SHIELDED WINDOWS. OVERHEAD MASTER/SLAVE MANIPULATORS (LEFT, ABOVE WORKING WINDOWS) WILL MOVE ACROSS GUIDE RAILS IN SLOT ABOVE THE WINDOWS. CAMERA FACING SOUTHEAST. INL NEGATIVE NO. 8996. Unknown Photographer, 10/28/1953 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  6. Hot electron plasmon-protected solar cell.

    PubMed

    Kong, J; Rose, A H; Yang, C; Wu, X; Merlo, J M; Burns, M J; Naughton, M J; Kempa, K

    2015-09-21

    A solar cell based on a hot electron plasmon protection effect is proposed and made plausible by simulations, non-local modeling of the response, and quantum mechanical calculations. In this cell, a thin-film, plasmonic metamaterial structure acts as both an efficient photon absorber in the visible frequency range and a plasmonic resonator in the IR range, the latter of which absorbs and protects against phonon emission the free energy of the hot electrons in an adjacent semiconductor junction. We show that in this structure, electron-plasmon scattering is much more efficient than electron-phonon scattering in cooling-off hot electrons, and the plasmon-stored energy is recoverable as an additional cell voltage. The proposed structure could become a prototype of a new generation of high efficiency solar cells.

  7. Radioactive hot cell access hole decontamination machine

    DOEpatents

    Simpson, William E.

    1982-01-01

    Radioactive hot cell access hole decontamination machine. A mobile housing has an opening large enough to encircle the access hole and has a shielding door, with a door opening and closing mechanism, for uncovering and covering the opening. The housing contains a shaft which has an apparatus for rotating the shaft and a device for independently translating the shaft from the housing through the opening and access hole into the hot cell chamber. A properly sized cylindrical pig containing wire brushes and cloth or other disks, with an arrangement for releasably attaching it to the end of the shaft, circumferentially cleans the access hole wall of radioactive contamination and thereafter detaches from the shaft to fall into the hot cell chamber.

  8. HOT CELL BUILDING, TRA632, INTERIOR. CONTEXTUAL VIEW OF HOT CELL ...

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

    HOT CELL BUILDING, TRA-632, INTERIOR. CONTEXTUAL VIEW OF HOT CELL NO. 2 FROM STAIRWAY ALONG NORTH WALL. OBSERVATION WINDOW ALONG WEST SIDE BENEATH "CELL 2" SIGN. DOORWAY IN LEFT OF VIEW LEADS TO CELL 1 WORK AREA OR TO EXIT OUTDOORS TO NORTH. RADIATION DETECTION MONITOR TO RIGHT OF DOOR. CAMERA FACING SOUTHWEST. INL NEGATIVE NO. HD46-28-3. Mike Crane, Photographer, 2/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  9. Dynamics of hot and dense nuclear and partonic matter

    SciTech Connect

    Bratkovskaya, E. L.; Cassing, W.; Linnyk, O.; Konchakovski, V. P.; Voronyuk, V.; Ozvenchuk, V.

    2012-06-15

    The dynamics of hot and dense nuclear matter is discussed from the microscopic transport point of view. The basic concepts of the Hadron-String-Dynamical transport model (HSD)-derived from Kadanoff-Baym equations in phase phase-are presented as well as 'highlights' of HSD results for different observables in heavy-ion collisions from 100 A MeV (SIS) to 21 A TeV(RHIC) energies. Furthermore, a novel extension of the HSD model for the description of the partonic phase-the Parton-Hadron-String-Dynamics (PHSD) approach-is introduced. PHSD includes a nontrivial partonic equation of state-in line with lattice QCD-as well as covariant transition rates from partonic to hadronic degrees of freedom. The sensitivity of hadronic observables to the partonic phase is demonstrated for relativistic heavy-ion collisions from the FAIR/NICA up to the RHIC energy regime.

  10. WESF hot cells waste minimization criteria hot cells window seals evaluation

    SciTech Connect

    Walterskirchen, K.M.

    1997-03-31

    WESF will decouple from B Plant in the near future. WESF is attempting to minimize the contaminated solid waste in their hot cells and utilize B Plant to receive the waste before decoupling. WESF wishes to determine the minimum amount of contaminated waste that must be removed in order to allow minimum maintenance of the hot cells when they are placed in ''laid-up'' configuration. The remaining waste should not cause unacceptable window seal deterioration for the remaining life of the hot cells. This report investigates and analyzes the seal conditions and hot cell history and concludes that WESF should remove existing point sources, replace cerium window seals in F-Cell and refurbish all leaded windows (except for A-Cell). Work should be accomplished as soon as possible and at least within the next three years.

  11. Isovector response function of hot nuclear matter with Skyrme interactions

    SciTech Connect

    Braghin, F.L.; Vautherin, D.; Abada, A.

    1995-11-01

    We investigate the role of the effective nucleon-nucleon interaction in the description of giant dipole resonances in hot nuclei. For this purpose we calculate the response function of hot nuclear matter to a small isovector external perturbation using various effective Skyrme interactions. We find that for Skyrme forces with an effective mass close to unity an undamped zero sound mode occurs at zero temperature. This mode gives rise in finite nuclei (calculated via the Steinwedel-Jenssen model) to a resonance whose energy agrees with the observed value. We find that zero sound disappears at a temperature of a few MeV, leaving only a broad peak in the dipole strength. For Skyrme forces with a small value of the effective mass (0.4), there is no zero sound at zero temperature but only a weak peak located too high in energy. The strength distribution in this case is nearly independent of temperature and shows small collective effects. The relevance of these results for the saturation of photon multiplicities observed in recent experiments is pointed out.

  12. Hot exciton dissociation in polymer solar cells.

    PubMed

    Grancini, G; Maiuri, M; Fazzi, D; Petrozza, A; Egelhaaf, H-J; Brida, D; Cerullo, G; Lanzani, G

    2013-01-01

    The standard picture of photovoltaic conversion in all-organic bulk heterojunction solar cells predicts that the initial excitation dissociates at the donor/acceptor interface after thermalization. Accordingly, on above-gap excitation, the excess photon energy is quickly lost by internal dissipation. Here we directly target the interfacial physics of an efficient low-bandgap polymer/PC(60)BM system. Exciton splitting occurs within the first 50 fs, creating both interfacial charge transfer states (CTSs) and polaron species. On high-energy excitation, higher-lying singlet states convert into hot interfacial CTSs that effectively contribute to free-polaron generation. We rationalize these findings in terms of a higher degree of delocalization of the hot CTSs with respect to the relaxed ones, which enhances the probability of charge dissociation in the first 200 fs. Thus, the hot CTS dissociation produces an overall increase in the charge generation yield.

  13. Hot photocarrier dynamics in organic solar cells.

    PubMed

    Lane, P A; Cunningham, P D; Melinger, J S; Esenturk, O; Heilweil, E J

    2015-07-16

    Photocurrent in an organic solar cell is generated by a charge transfer reaction between electron donors and acceptors. Charge transfer is expected to proceed from thermalized states, but this picture has been challenged by recent studies that have investigated the role of hot excitons. Here we show a direct link between excess excitation energy and photocarrier mobility. Charge transfer from excited donor molecules generates hot photocarriers with excess energy coming from the offset between the lowest unoccupied molecular orbital of the donor and that of the acceptor. Hot photocarriers manifest themselves through a short-lived spike in terahertz photoconductivity that decays on a picosecond timescale as carriers thermalize. Different dynamics are observed when exciting the acceptor at its absorption edge to a thermalized state. Charge transfer in this case generates thermalized carriers described by terahertz photoconductivity dynamics consisting of an instrument-limited rise to a long-lived signal.

  14. 1. View of rmad from jr. hot cell, facing north ...

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

    1. View of r-mad from jr. hot cell, facing north - Nevada Test Site, Reactor Maintenance & Disassembly Complex, Junior Hot Cell, Jackass Flats, Area 25, South of intersection of Roads F & G, Mercury, Nye County, NV

  15. HOT CELL BUILDING, TRA632, INTERIOR. HOT CELL NO. 1 (THE ...

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

    HOT CELL BUILDING, TRA-632, INTERIOR. HOT CELL NO. 1 (THE FIRST BUILT) IN LABORATORY 101. CAMERA FACES SOUTHEAST. SHIELDED OPERATING WINDOWS ARE ON LEFT (NORTH) SIDE. OBSERVATION WINDOW IS AT LEFT OF VIEW (ON WEST SIDE). PLASTIC COVERS SHROUD MASTER/SLAVE MANIPULATORS AT WINDOWS IN LEFT OF VIEW. NOTE MINERAL OIL RESERVOIR ABOVE "CELL 1" SIGN, INDICATING LEVEL OF THE FLUID INSIDE THE THICK WINDOWS. HOT CELL HAS BEVELED CORNER BECAUSE A SQUARED CORNER WOULD HAVE SUPPLIED UNNECESSARY SHIELDING. NOTE PUMICE BLOCK WALL AT LEFT OF VIEW. INL NEGATIVE NO. HD46-28-1. Mike Crane, Photographer, 2/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  16. Functional study of hot pepper 26S proteasome subunit RPN7 induced by Tobacco mosaic virus from nuclear proteome analysis

    SciTech Connect

    Lee, Boo-Ja; Kwon, Sun Jae; Kim, Sung-Kyu; Kim, Ki-Jeong; Park, Chang-Jin; Kim, Young-Jin; Park, Ohkmae K.; Paek, Kyung-Hee . E-mail: khpaek95@korea.ac.kr

    2006-12-15

    Two-dimensional gel electrophoresis (2-DE) was applied for the screening of Tobacco mosaic virus (TMV)-induced hot pepper (Capsicum annuum cv. Bugang) nuclear proteins. From differentially expressed protein spots, we acquired the matched peptide mass fingerprint (PMF) data, analyzed by MALDI-TOF MS, from the non-redundant hot pepper EST protein FASTA database using the VEMS 2.0 software. Among six identified nuclear proteins, the hot pepper 26S proteasome subunit RPN7 (CaRPN7) was subjected to further study. The level of CaRPN7 mRNA was specifically increased during incompatible TMV-P{sub 0} interaction, but not during compatible TMV-P{sub 1.2} interaction. When CaRPN7::GFP fusion protein was targeted in onion cells, the nuclei had been broken into pieces. In the hot pepper leaves, cell death was exacerbated and genomic DNA laddering was induced by Agrobacterium-mediated transient overexpression of CaPRN7. Thus, this report presents that the TMV-induced CaRPN7 may be involved in programmed cell death (PCD) in the hot pepper plant.

  17. Functional study of hot pepper 26S proteasome subunit RPN7 induced by Tobacco mosaic virus from nuclear proteome analysis.

    PubMed

    Lee, Boo-Ja; Kwon, Sun Jae; Kim, Sung-Kyu; Kim, Ki-Jeong; Park, Chang-Jin; Kim, Young-Jin; Park, Ohkmae K; Paek, Kyung-Hee

    2006-12-15

    Two-dimensional gel electrophoresis (2-DE) was applied for the screening of Tobacco mosaic virus (TMV)-induced hot pepper (Capsicum annuum cv. Bugang) nuclear proteins. From differentially expressed protein spots, we acquired the matched peptide mass fingerprint (PMF) data, analyzed by MALDI-TOF MS, from the non-redundant hot pepper EST protein FASTA database using the VEMS 2.0 software. Among six identified nuclear proteins, the hot pepper 26S proteasome subunit RPN7 (CaRPN7) was subjected to further study. The level of CaRPN7 mRNA was specifically increased during incompatible TMV-P(0) interaction, but not during compatible TMV-P(1.2) interaction. When CaRPN7::GFP fusion protein was targeted in onion cells, the nuclei had been broken into pieces. In the hot pepper leaves, cell death was exacerbated and genomic DNA laddering was induced by Agrobacterium-mediated transient overexpression of CaPRN7. Thus, this report presents that the TMV-induced CaRPN7 may be involved in programmed cell death (PCD) in the hot pepper plant.

  18. HOT CELL BUILDING, TRA632, INTERIOR. DETAIL OF HOT CELL NO. ...

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

    HOT CELL BUILDING, TRA-632, INTERIOR. DETAIL OF HOT CELL NO. 2 SHOWS MANIPULATION INSTRUMENTS AND SHIELDED OPERATING WINDOWS. PENETRATIONS FOR OPERATING INSTRUMENTS GO THROUGH SHIELDING ABOVE WINDOWS. CONDUIT FOR UTILITIES AND CONTROLS IS BEHIND METAL CABINET BELOW WINDOWS NEAR FLOOR. CAMERA FACES WEST. WARNING SIGN LIMITS FISSILE MATERIAL TO SPECIFIED NUMBER OF GRAMS OF URANIUM AND PLUTONIUM. INL NEGATIVE NO. HD46-28-2. Mike Crane, Photographer, 2/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  19. HOT CELL BUILDING, TRA632. CONTEXTUAL AERIAL VIEW OF HOT CELL ...

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

    HOT CELL BUILDING, TRA-632. CONTEXTUAL AERIAL VIEW OF HOT CELL BUILDING, IN VIEW AT LEFT, AS YET WITHOUT ROOF. PLUG STORAGE BUILDING LIES BETWEEN IT AND THE SOUTH SIDE OF THE MTR BUILDING AND ITS WING. NOTE CONCRETE DRIVE BETWEEN ROLL-UP DOOR IN MTR BUILDING AND CHARGING FACE OF PLUG STORAGE. REACTOR SERVICES BUILDING (TRA-635) WILL COVER THIS DRIVE AND BUTT UP TO CHARGING FACE. DOTTED LINE IS ON ORIGINAL NEGATIVE. TRA PARKING LOT IN LEFT CORNER OF THE VIEW. CAMERA FACING NORTHWESTERLY. INL NEGATIVE NO. 8274. Unknown Photographer, 7/2/1953 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  20. Hot cell shield plug extraction apparatus

    DOEpatents

    Knapp, Philip A.; Manhart, Larry K.

    1995-01-01

    An apparatus is provided for moving shielding plugs into and out of holes in concrete shielding walls in hot cells for handling radioactive materials without the use of external moving equipment. The apparatus provides a means whereby a shield plug is extracted from its hole and then swung approximately 90 degrees out of the way so that the hole may be accessed. The apparatus uses hinges to slide the plug in and out and to rotate it out of the way, the hinge apparatus also supporting the weight of the plug in all positions, with the load of the plug being transferred to a vertical wall by means of a bolting arrangement.

  1. Handling of Highly Radioactive Radiation Sources in a Hot Cell Using a Mechanically Driven Cell Crane - 13452

    SciTech Connect

    Klute, Stefan; Huber, Wolfgang-Bruno

    2013-07-01

    In 2010, Siempelkamp Nukleartechnik GmbH was awarded the contract for design and erection of a Hot Cell for handling and storage of highly radioactive radiation sources. This Hot Cell is part of a new hot cell laboratory, constructed for the NHZ (Neues Handhabungszentrum = New Handling Center) of the Nuclear Engineering Seibersdorf GmbH (NES). All incurring radioactive materials from Austria are collected in the NHZ, where they are safely conditioned and stored temporarily until their final storage. The main tasks of the NES include, apart from the collection, conditioning and storage of radioactive waste, also the reprocessing and the decontamination of facilities and laboratories originating from 45 years of research and development at the Seibersdorf site as well as the operation of the Hot Cell Laboratory [1]. The new Hot Cell Laboratory inside the NHZ consists of the following room areas: - One hot cell, placed in the center, for remote controlled, radiation protected handling of radioactive materials, including an integrated floor storage for the long-term temporary storage of highly radioactive radiation sources; - An anteroom for the loading and unloading of the hot cell; - One control room for the remote controlling of the hot cell equipment; - One floor storage, placed laterally to the hot cell, for burial, interim storage and removal of fissionable radioactive material in leak-proof packed units in 100 l drums. The specific design activity of the hot cell of 1.85 Pbq relating to 1-Me-Radiator including the integrated floor storage influences realization and design of the components used in the cell significantly. (authors)

  2. Hot cell facility design for large fusion devices

    SciTech Connect

    Barrett, R.J.; Bussell, G.T.

    1985-01-01

    Large hot cell facilities will be necessary to support the operation of large fusion devices. The supporting hot cells will be needed to serve a variety of different functions and tasks, which include reactor component maintenance, tool and maintenance equipment repair, and preparation of radioactive material for shipment and disposal. This paper discusses hot cell facility functions, requirements, and design issues and techniques. Suggested solutions and examples are given.

  3. Decontamination of Hot Cells and Hot Pipe Tunnel at NASA's Plum Brook Reactor Facility

    SciTech Connect

    Anderson, M.G.; Halishak, W.F.

    2008-07-01

    The large scale decontamination of the concrete Hot Cells and Hot Pipe Tunnel at NASA's Plum Brook Reactor Facility demonstrates that novel management and innovative methods are crucial to ensuring that the successful remediation of the most contaminated facilities can be achieved with minimal risk to the project stakeholders. (authors)

  4. Verification survey of buildings 200 hot cells

    SciTech Connect

    Sholeen, C.M.

    1996-03-01

    At the start of this D&D project, the decontamination goals were set at (1) reducing the stack emissions to 10% of the 1991 emissions; (2) reducing the exposure rate in each cell to < 1 mR/h; and (3) reducing the removable contamination to none detectable. Since the contamination can be fixed with paint, the other two goals were given priority. The estimate of the 1995 emissions from K-3 was 20% of the 1991 emissions estimate. However, the 1996 estimates are {approximately}9% of the 1991 emissions estimate. Since in 1991 the K-3 emissions were only 1/2% of the emissions from M-1, even the 20% reduction has little effect on the project reduction. The total emissions have been reduce to {approximately}2 1/4% of the 1991 emissions from the 5 hot cells that were decontaminated. The emissions and exposure rates are presented in Table I below. Cells A-1 and M-1 exceed the exposure rate criteria. For the other cells, the general exposure rate in the middle of the cell meets the criteria. However, near the prefilters, the exposure rates increase. Cell M-1 has extensive floor contamination that penetrated to a 6 inch depth. At 30 cm above the floor, the exposure rate through the lead blanket is 50 mR/h. A more detailed list of acceptance criteria were specified before the final verification survey. Table ii compares the maximum survey results on the wall or floor surface of each cell to these criteria. Cells M-1 and A-1 frequently fail to meet these criteria.

  5. HOT CELL BUILDING, TRA632. FIRST FLOOR FOUNDATION PLAN SHOWS SECTIONALIZED ...

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

    HOT CELL BUILDING, TRA-632. FIRST FLOOR FOUNDATION PLAN SHOWS SECTIONALIZED FLOOR LOADINGS AND CONCRETE SLAB THICKNESSES, A TYPICAL FEATURE OF NUCLEAR ARCHITECTURE. IDAHO OPERATIONS OFFICE MTR-632-IDO-2, 11/1952. INL INDEX NO. 531-0632-62-396-110561, REV. 1. - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  6. Bulk processing of radionuclide generator parents at the Los Alamos Hot Cell Facility

    SciTech Connect

    Fassbender, M. E.; Nortier, F. M.; Phillips, Dennis R.; Peterson, E. J.

    2004-01-01

    Bulk radionuclide processing at Los Alamos includes isotopes with short-lived radioactive daughter nuclides ('generator parents') for medical applications. The generator radionuclide parents {sup 68}Ge, {sup 82}Sr, {sup 88}Zr and {sup 109}Cd are regularly processed at the Los Alamos Hot Cell Facility. Nuclear chemical aspects related to the production and processing of these generator parents are briefly outlined.

  7. 47. ARAI. Interior view of operating wall of hot cell ...

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

    47. ARA-I. Interior view of operating wall of hot cell in ARA-626. Note stands for operators at viewing windows. Manipulators with hand grips extend cables and other controls into hot cell through ducts above windows. Ineel photo no. 81-27. - Idaho National Engineering Laboratory, Army Reactors Experimental Area, Scoville, Butte County, ID

  8. 15. View of interior, north wall of hot cell featuring ...

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

    15. View of interior, north wall of hot cell featuring radioactive materials containment box, facing east - Nevada Test Site, Reactor Maintenance & Disassembly Complex, Junior Hot Cell, Jackass Flats, Area 25, South of intersection of Roads F & G, Mercury, Nye County, NV

  9. Hanford spent nuclear fuel hot conditioning system test procedure

    SciTech Connect

    Cleveland, K.J.

    1997-09-16

    This document provides the test procedures for cold testing of the prototype Hot Conditioning System (HCS) at the 306E Facility. The primary objective of this testing is to confirm design choices and provide data for the detailed design package prior to procurement of the process equipment. The current scope of testing in this document includes a fabricability study of the HCS, equipment performance testing of the HCS components, heat-up and cool-down cycle simulation, and robotic arm testing.

  10. Hot startup experience with electrometallurgical treatment of spent nuclear fuel

    SciTech Connect

    Benedict, R.W.; Lineberry, M.J.; McFarlane, H.F.; Rigg, R.H.

    1997-10-01

    The treatment of spent metal fuel from the EBR-II fast reactor commenced in June of 1996 at the Fuel Conditioning Facility on the Argonne-West site in Idaho, USA. During the first year of hot operations, 20 fuel assemblies entered processing and 6 low enrichment uranium product ingots were produced. Results are presented for the various process steps with decontamination factors achieved and equipment operational history reported.

  11. The "hot nose" sign on brain death nuclear scintigraphy: where does the flow really go?

    PubMed

    Appelt, Eric A; Song, Won S; Phillips, William T; Metter, Darlene F; Salman, Umber A; Blumhardt, Ralph

    2008-01-01

    Nuclear scintigraphy has been used in patients with brain death since the 1970s. Many studies report a "hot nose" sign as predictive of brain death and lack of cerebral flow. Current nuclear medicine textbooks state that increased flow to the nose occurs secondary to occlusion of the internal carotid artery with flow rerouted to the nose via the external carotid artery. This explanation has been provided for decades assuming that the blood flow is actually increased to the nose. We performed a study to determine whether flow is really seen in the nose when a hot nose sign is present.

  12. Human factors evaluation of the Auxiliary Hot Cell Facility, Sandia National Laboratories, Albuquerque, New Mexico.

    SciTech Connect

    Hunter, Regina Lee; Whitehurst, Hugh O.

    2003-11-01

    The Auxiliary Hot Cell Facility (AHCF) at Sandia National Laboratories, New Mexico (SNL/NM) is a Hazard Category 3 nuclear facility used to characterize, treat, and repackage radioactive and mixed material for reuse, recycling, or ultimate disposal. Mixed waste may also be handled at the AHCF. A significant upgrade to a previous facility, the Temporary Hot Cell, was required to perform this mission. A checklist procedure was used to perform a human-factors evaluation of the AHCF modifications. This evaluation resulted in two recommendations, both of which have been implemented.

  13. Bovine somatic cell nuclear transfer.

    PubMed

    Ross, Pablo J; Cibelli, Jose B

    2010-01-01

    Somatic cell nuclear transfer (SCNT) is a technique by which the nucleus of a differentiated cell is introduced into an oocyte from which its genetic material has been removed by a process called enucleation. In mammals, the reconstructed embryo is artificially induced to initiate embryonic development (activation). The oocyte turns the somatic cell nucleus into an embryonic nucleus. This process is called nuclear reprogramming and involves an important change of cell fate, by which the somatic cell nucleus becomes capable of generating all the cell types required for the formation of a new individual, including extraembryonic tissues. Therefore, after transfer of a cloned embryo to a surrogate mother, an offspring genetically identical to the animal from which the somatic cells where isolated, is born. Cloning by nuclear transfer has potential applications in agriculture and biomedicine, but is limited by low efficiency. Cattle were the second mammalian species to be cloned after Dolly the sheep, and it is probably the most widely used species for SCNT experiments. This is, in part due to the high availability of bovine oocytes and the relatively higher efficiency levels usually obtained in cattle. Given the wide utilization of this species for cloning, several alternatives to this basic protocol can be found in the literature. Here we describe a basic protocol for bovine SCNT currently being used in our laboratory, which is amenable for the use of the nuclear transplantation technique for research or commercial purposes.

  14. Equipment design guidelines for remote hot cell operations.

    SciTech Connect

    Wahlquist, D. R.

    1998-07-10

    Hot cells provide a unique and challenging environment for designing remotely operated equipment. A typical hot cell is an isolated room used to protect operators from highly contaminated and radioactive equipment. Hot cells usually have thick reinforced concrete walk and leaded glass windows. Operations within the hot cell are accomplished using master-slave manipulators and overhead crane or electro-mechanical manipulator systems. The inability to perform hands-on operation and maintenance in hot cells requires special design considerations. Some of these design considerations include operational interfaces, radiation, accessibility, replaceability/interchangeability, decontamination, atmospheric conditions, functionality, operator fatigue, and ease of use. This paper will discuss guidelines for designing hot cell remotely operated equipment that has been used successfully at Argonne National Laboratory. General topics in this paper will include master-slave manipulator types and limitations, overhead handling systems, viewing limitations, types and sizes of typical fasteners, hot cell compatible materials, mockup testing, guide features for mating parts, modularity, labeling, electrical fasteners, and lifting fixtures.

  15. Characterization report for Building 301 Hot Cell Facility

    SciTech Connect

    1998-07-01

    During the period from October, 1997, through March, 1998, ANL-E Health Physics conducted a pre-D and D characterization of Building 301, referred to as the Hot Cell Facility. While primary emphasis was placed on radiological evaluation, the presence of non-nuclear hazardous and toxic material was also included in the scope of the characterization. This is one of the early buildings on the ANL-E site, and was heavily used in the 1950`s and 1960`s for various nuclear reaction and reactor design studies. Some degree of cleanup and contamination fixation was done in the 1970`s, so that the building could be used with a minimum of risk of personnel contamination. Work records are largely nonexistent for the early history of the building, so that any assumptions about extent and type of contamination had to be kept very open in the survey planning process. The primary contaminant was found to be painted-over Cs-137 embedded in the concrete floors, although a variety of other nuclides consistent with the work said to have been performed were found in smaller quantities. Due to leaks and drips through the floor, a relatively modest amount of soil contamination was found in the service trench under the building, not penetrating deeply. Two contaminated, disconnected drain lines leaving the building could not be traced by site records, and remain a problem for remediation. The D and D Characterization Plan was fulfilled.

  16. Assessment of a hot hydrogen nuclear propulsion fuel test facility

    SciTech Connect

    Watanabe, Harry H.; Howe, Steven D.; Wantuck, Paul J.

    1991-01-01

    Subsequent to the announcement of the Space Exploration Initiative (SEI), several studies and review groups have identified nuclear thermal propulsion as a high priority technology for development. To achieve the goals of SEI to place man on Mars, a nuclear rocket will operate at near 2700K and in a hydrogen environment at near 60 atmospheres. Under these conditions, the operational lifetime of the rocket will be limited by the corrosion rate at the hydrogen/fuel interface. Consequently, the Los Alamos National Laboratory has been evaluating requirements and design issues for a test facility. The facility will be able to directly heat fuel samples by electrical resistance, microwave deposition, or radio frequency induction heating to temperatures near 3000K. Hydrogen gas at variable pressure and temperatures will flow through the samples. The thermal gradients, power density, and operating times envisioned for nuclear rockets will be duplicated as close as reasonable. The post-sample flow stream will then be scrubbed and cooled before reprocessing. The baseline design and timetable for the facility will be discussed. 7 refs.

  17. Analysis of Material Sample Heated by Impinging Hot Hydrogen Jet in a Non-Nuclear Tester

    NASA Technical Reports Server (NTRS)

    Wang, Ten-See; Foote, John; Litchford, Ron

    2006-01-01

    A computational conjugate heat transfer methodology was developed and anchored with data obtained from a hot-hydrogen jet heated, non-nuclear materials tester, as a first step towards developing an efficient and accurate multiphysics, thermo-fluid computational methodology to predict environments for hypothetical solid-core, nuclear thermal engine thrust chamber. The computational methodology is based on a multidimensional, finite-volume, turbulent, chemically reacting, thermally radiating, unstructured-grid, and pressure-based formulation. The multiphysics invoked in this study include hydrogen dissociation kinetics and thermodynamics, turbulent flow, convective and thermal radiative, and conjugate heat transfers. Predicted hot hydrogen jet and material surface temperatures were compared with those of measurement. Predicted solid temperatures were compared with those obtained with a standard heat transfer code. The interrogation of physics revealed that reactions of hydrogen dissociation and recombination are highly correlated with local temperature and are necessary for accurate prediction of the hot-hydrogen jet temperature.

  18. Nuclear fusion in the deuterated cores of inflated hot Jupiters

    NASA Astrophysics Data System (ADS)

    Ouyed, Rachid; Jaikumar, Prashanth

    2016-03-01

    Ouyed et al. (Astrophys. J. 501:367, 1998) proposed Deuterium (DD) fusion at the core-mantle interface of giant planets as a mechanism to explain their observed heat excess. But rather high interior temperatures (˜105 K) and a stratified D layer are needed, making such a scenario unlikely. In this paper, we re-examine DD fusion, with the addition of screening effects pertinent to a deuterated core containing ice and some heavy elements. This alleviates the extreme temperature constraint and removes the requirement of a stratified D layer. As an application, we propose that, if their core temperatures are a few times 104 K and core composition is chemically inhomogeneous, the observed inflated size of some giant exoplanets ("hot Jupiters") may be linked to screened DD fusion occurring deep in the interior. Application of an analytic evolution model suggests that the amount of inflation from this effect can be important if there is sufficient rock-ice in the core, making DD fusion an effective extra internal energy source for radius inflation. The mechanism of screened DD fusion, operating in the above temperature range, is generally consistent with the trend in radius anomaly with planetary equilibrium temperature T_{eq}, and also depends on planetary mass. Although we do not consider the effect of incident stellar flux, we expect that a minimum level of irradiation is necessary to trigger core erosion and subsequent DD fusion inside the planet. Since DD fusion is quite sensitive to the screening potential inferred from laboratory experiments, observations of inflated hot Jupiters may help constrain screening effects in the cores of giant planets.

  19. Preliminary safety analysis report for the Auxiliary Hot Cell Facility, Sandia National Laboratories, Albuquerque, New Mexico

    SciTech Connect

    OSCAR,DEBBY S.; WALKER,SHARON ANN; HUNTER,REGINA LEE; WALKER,CHERYL A.

    1999-12-01

    The Auxiliary Hot Cell Facility (AHCF) at Sandia National Laboratories, New Mexico (SNL/NM) will be a Hazard Category 3 nuclear facility used to characterize, treat, and repackage radioactive and mixed material and waste for reuse, recycling, or ultimate disposal. A significant upgrade to a previous facility, the Temporary Hot Cell, will be implemented to perform this mission. The following major features will be added: a permanent shield wall; eight floor silos; new roof portals in the hot-cell roof; an upgraded ventilation system; and upgraded hot-cell jib crane; and video cameras to record operations and facilitate remote-handled operations. No safety-class systems, structures, and components will be present in the AHCF. There will be five safety-significant SSCs: hot cell structure, permanent shield wall, shield plugs, ventilation system, and HEPA filters. The type and quantity of radionuclides that could be located in the AHCF are defined primarily by SNL/NM's legacy materials, which include radioactive, transuranic, and mixed waste. The risk to the public or the environment presented by the AHCF is minor due to the inventory limitations of the Hazard Category 3 classification. Potential doses at the exclusion boundary are well below the evaluation guidelines of 25 rem. Potential for worker exposure is limited by the passive design features incorporated in the AHCF and by SNL's radiation protection program. There is no potential for exposure of the public to chemical hazards above the Emergency Response Protection Guidelines Level 2.

  20. Extended Characterization of Chemical Processes in Hot Cells Using Environmental Swipe Samples

    SciTech Connect

    Olsen, Khris B.; Mitroshkov, Alexandre V.; Thomas, M-L; Lepel, Elwood A.; Brunson, Ronald R.; Ladd-Lively, Jennifer

    2012-09-15

    Environmental sampling is used extensively by the International Atomic Energy Agency (IAEA) for verification of information from State declarations or a facility’s design regarding nuclear activities occurring within the country or a specific facility. Environmental sampling of hot cells within a facility under safeguards is conducted using 10.2 cm x 10.2 cm cotton swipe material or cellulose swipes. Traditional target analytes used by the IAEA to verify operations within a facility include a select list of gamma-emitting radionuclides and total and isotopic U and Pu. Analysis of environmental swipe samples collected within a hot-cell facility where chemical processing occurs may also provide information regarding specific chemicals used in fuel processing. However, using swipe material to elucidate what specific chemical processes were/are being used within a hot cell has not been previously evaluated. Staff from Pacific Northwest National Laboratory (PNNL) and Oak Ridge National Laboratory (ORNL) teamed to evaluate the potential use of environmental swipe samples as collection media for volatile and semivolatile organic compounds. This evaluation was initiated with sample collection during a series of Coupled End-to-End (CETE) reprocessing runs at ORNL. The study included measurement of gamma emitting radionuclides, total and isotopic U and Pu, and volatile and semivolatile organic compounds. These results allowed us to elucidate what chemical processes used in the hot cells during reprocessing of power reactor and identify other legacy chemicals used in hot cell operations which predate the CETE process.

  1. Formation of hot particles during the Chernobyl nuclear power plant accident

    SciTech Connect

    Kashparov, V.A.; Ivanov, Y.A.; Zvarisch, S.I.; Protsak, V.P.; Khomutinin, Y.V.; Kurepin, A.D.; Pazukhin, E.M.

    1996-05-01

    The oxidation of irradiated Chernobyl nuclear fuel at 670 to 1,170 K for 3 to 21 h resulted in its destruction into fine particles, the dispersal composition of which is well described by lognormal distribution regularity. The median radius of the formed particles does not depend on the annealing temperature and decreases with the increase of the annealing period from 10 to 3 {micro}m. Proceeding from the dispersal composition and matrix composition of the Chernobyl hot fuel particles, it can be concluded that the oxidation of nuclear fuel was one of the basic mechanisms of hot fuel particle formation during the accident at the Chernobyl nuclear power plant. With oxidation in air and the dispersal of irradiated oxide nuclear fuel at as low as 670 K, ruthenium, located on the granular borders, is released. Ruthenium is oxidized to volatile RuO{sub 4}, sublimated, and condensed on materials of iron. Nickel and stainless steel can be efficiently used at high temperatures (tested to 1,200 K) for radioruthenium adsorption in accidents and for some technological operations. As the temperature of hot fuel particles annealed in inert media increases from 1,270 to 2,270 K, the relative release of radionuclides increases in the following sequence: cesium isotopes; europium isotopes; cerium isotopes; americium isotopes; and ruthenium, plutonium, and curium isotopes.

  2. K Basin spent nuclear fuel hot conditioning system functions {ampersand} requirements

    SciTech Connect

    Miska, C.R., Westinghouse Hanford

    1996-07-08

    The purpose of this F{ampersand}R document is to establish the functional requirements baseline for the Spent Nuclear Fuel Hot Conditioning System (HCS) subproject. This F{ampersand}R documents the: -mission of the HCS, -evolution of the technical baseline leading to the HCS, -functions that must be performed to accomplish the HCS mission, -requirements basis allocated to the HCS mission and functions, -identification and definition of interfaces between the HCS and other SNF subprojects.

  3. {open_quote}{open_quote}Hot spots{close_quote}{close_quote} effect in nuclear shadowing

    SciTech Connect

    Zhu, W.; Ruan, J. |

    1996-07-01

    Shadowing in inelastic lepton-nucleus scattering is analyzed by using the modified Gribov-Levin-Ryskin evolution equation. We find that a comparison of the structure functions between nuclear and proton targets at {ital x}{lt}10{sup {minus}3} and {ital Q}{sup 2}{approx_gt}1 GeV{sup 2} can provide useful information about the hot spots of the nucleon. {copyright} {ital 1996 The American Physical Society.}

  4. Fundamental Limitations to Plasmonic Hot-Carrier Solar Cells.

    PubMed

    Zhang, Yu; Yam, ChiYung; Schatz, George C

    2016-05-19

    Detailed balance between photon-absorption and energy loss constrains the efficiency of conventional solar cells to the Shockley-Queisser limit. However, if solar illumination can be absorbed over a wide spectrum by plasmonic structures, and the generated hot-carriers can be collected before relaxation, the efficiency of solar cells may be greatly improved. In this work, we explore the opportunities and limitations for making plasmonic solar cells, here considering a design for hot-carrier solar cells in which a conventional semiconductor heterojunction is attached to a plasmonic medium such as arrays of gold nanoparticles. The underlying mechanisms and fundamental limitations of this cell are studied using a nonequilibrium Green's function method, and the numerical results indicate that this cell can significantly improve the absorption of solar radiation without reducing open-circuit voltage, as photons can be absorbed to produce mobile carriers in the semiconductor as long as they have energy larger than the Schottky barrier rather than above the bandgap. However, a significant fraction of the hot-carriers have energies below the Schottky barrier, which makes the cell suffer low internal quantum efficiency. Moreover, quantum efficiency is also limited by hot-carrier relaxation and metal-semiconductor coupling. The connection of these results to recent experiments is described, showing why plasmonic solar cells can have less than 1% efficiency.

  5. Fundamental limitations of hot-carrier solar cells

    NASA Astrophysics Data System (ADS)

    Kirk, A. P.; Fischetti, M. V.

    2012-10-01

    Sunlight-generated hot-carrier transport in strongly absorbing direct band-gap GaAs—among the most optimal of semiconductors for high-efficiency solar cells—is simulated with an accurate full-band structure self-consistent Monte Carlo method, including short- and long-range Coulomb interaction, impact ionization, and optical and acoustic phonon scattering. We consider an ultrapure 100-nm-thick intrinsic GaAs absorber layer designed with quasiballistic carrier transport that achieves complete photon absorption down to the band edge by application of careful light trapping and that has a generous hot-carrier retention time of 10 ps prior to the onset of carrier relaxation. We find that hot-carrier solar cells can be severely limited in performance due to the substantially reduced current density caused by insufficient extraction of the widely distributed hot electrons (holes) through the requisite energy selective contacts.

  6. The Hanford spent nuclear metal fuel multi-canister overpack and vacuum drying & hot conditioning process

    SciTech Connect

    Goldmann, L.H.; Irwin, J.J.; Miska, C.R.

    1996-12-31

    Nuclear production reactors operated at the U.S. Department of Energy`s Hanford Site from 1944 until 1988 to produce plutonium. Most of the irradiated fuel from these reactors was processed onsite to separate and recover the plutonium. When the processing facilities were closed in 1992, about 1,900 metric tons of unprocessed irradiated fuel remained in storage. Additional fuel was irradiated for research purposes or was shipped to the Hanford Site from offsite reactor facilities for storage or recovery of nuclear materials. The fuel inventory now in storage at the Hanford Site is predominantly N Reactor irradiated fuel, a metallic uranium alloy that is coextruded into zircaloy-2 cladding. The Spent Nuclear Fuel Project has committed to an accelerated schedule for removing spent nuclear fuel from the Hanford Site K Basins to a new interim storage facility in the 200 Area. The Westinghouse Hanford Company has developed an integrated process to deal with the K Basin spent fuel inventory. The process consists of cleaning the fuel, packaging it into MCOs, vacuum drying it at the K Basins, then transporting it to the Canister Storage Building for staging, hot conditioning, and interim storage. This presentation describes the MCO function, design, and life-cycle, including an overview of the vacuum drying and hot conditioning processes.

  7. Nuclear choreography: interpretations from living cells.

    PubMed

    Janicki, Susan M; Spector, David L

    2003-04-01

    The advent of green fluorescent protein technology, its use in photobleaching experiments and the development of methods to rapidly acquire images and analyze complex datasets have opened the door to unraveling the mechanisms of nuclear functions in living cells. Studies over the past few years have characterized the movement of chromatin, nuclear proteins and nuclear bodies and, in some cases, correlated their dynamics with energy dependence, cell cycle progression, developmental changes, factor targeting and nuclear position. The mechanisms by which nuclear components move or are restrained have important implications for understanding not only the efficacy of nuclear functions but also the regulation of developmental programs and cellular growth.

  8. MAGNETICALLY CONFINED INTERSTELLAR HOT PLASMA IN THE NUCLEAR BULGE OF OUR GALAXY

    SciTech Connect

    Nishiyama, Shogo; Kwon, Jungmi; Tamura, Motohide; Yasui, Kazuki; Nagata, Tetsuya; Yoshikawa, Tatsuhito; Uchiyama, Hideki; Schödel, Rainer; Hatano, Hirofumi; Sato, Shuji; Sugitani, Koji; Suenaga, Takuya

    2013-06-01

    The origin of the Galactic center diffuse X-ray emission (GCDX) is still under intense investigation. In particular, the interpretation of the hot (kT ≈ 7 keV) component of the GCDX, characterized by the strong Fe 6.7 keV line emission, has been contentious. If the hot component originates from a truly diffuse interstellar plasma, not a collection of unresolved point sources, such plasma cannot be gravitationally bound, and its regeneration would require a huge amount of energy. Here, we show that the spatial distribution of the GCDX does not correlate with the number density distribution of an old stellar population traced by near-infrared light, strongly suggesting a significant contribution of the diffuse interstellar plasma. Contributions of the old stellar population to the GCDX are implied to be ∼50% and ∼20% in the nuclear stellar disk (NSD) and nuclear star cluster, respectively. For the NSD, a scale height of 0.°32 ± 0.°02 is obtained for the first time from the stellar number density profiles. We also show the results of the extended near-infrared polarimetric observations in the central 3° × 2° region of our Galaxy, and confirm that the GCDX region is permeated by a large scale, toroidal magnetic field (MF) as previously claimed. Together with observed MF strengths close to energy equipartition, the hot plasma could be magnetically confined, reducing the amount of energy required to sustain it.

  9. Nuclear Mechanics and Stem Cell Differentiation.

    PubMed

    Mao, Xinjian; Gavara, Nuria; Song, Guanbin

    2015-12-01

    Stem cells are characterized by their self-renewal and multi-lineage differentiation potential. Stem cell differentiation is a prerequisite for the application of stem cells in regenerative medicine and clinical therapy. In addition to chemical stimulation, mechanical cues play a significant role in regulating stem cell differentiation. The integrity of mechanical sensors is necessary for the ability of cells to respond to mechanical signals. The nucleus, the largest and stiffest cellular organelle, interacts with the cytoskeleton as a key mediator of cell mechanics. Nuclear mechanics are involved in the complicated interactions of lamins, chromatin and nucleoskeleton-related proteins. Thus, stem cell differentiation is intimately associated with nuclear mechanics due to its indispensable role in mechanotransduction and mechanical response. This paper reviews several main contributions of nuclear mechanics, highlights the hallmarks of the nuclear mechanics of stem cells, and provides insight into the relationship between nuclear mechanics and stem cell differentiation, which may guide clinical applications in the future.

  10. Sampling Tools for Contaminated Hot Cells and Remotely Accessed Tanks

    SciTech Connect

    Rostomashvili, Z.; Chabashvili, Mariam; Tingey, Joel M.; Gauerke, Eric R.; Szytel, Lisa; Pires, Richard P.; O'Connor, William

    2015-07-30

    The U.S. Department of Energy (DOE) and the Pacific Northwest National Laboratory in collaboration with the Andronikashvili Institute of Physics in Tbilisi, Georgia have developed and tested technologies which sample contaminated areas inside hot cell facilities. Sampling locations in the facility include drains, ventilation exhaust ducts, sumps and shielded waste tanks. Tool designs for sampling each location include the ability to introduce the tool using existing ports, pass-throughs or inter-cells trolley and operate the tool with the existing remote manipulators. Areas inside the hot cell facility that would likely be contaminated from chemically processing irradiated reactor targets and fuel even after decontamination have been identified as potential sampling locations. Andronikashvili Institute of Physics in Tbilisi, Georgia has access to a reactor, hot cells, glove boxes and storage facilities that have been used to develop and test hot cell sampling technologies. These facilities are uncontaminated with radiological material and provide the ability to test these sampling technologies on representative equipment without contaminating the tools. Prototype tools have been fabricate and tested at these facilities using simulants representative of radiological contamination and waste.

  11. Signals of Bose Einstein condensation and Fermi quenching in the decay of hot nuclear systems

    NASA Astrophysics Data System (ADS)

    Marini, P.; Zheng, H.; Boisjoli, M.; Verde, G.; Chbihi, A.; Napolitani, P.; Ademard, G.; Augey, L.; Bhattacharya, C.; Borderie, B.; Bougault, R.; Frankland, J. D.; Fable, Q.; Galichet, E.; Gruyer, D.; Kundu, S.; La Commara, M.; Lombardo, I.; Lopez, O.; Mukherjee, G.; Parlog, M.; Rivet, M. F.; Rosato, E.; Roy, R.; Spadaccini, G.; Vigilante, M.; Wigg, P. C.; Bonasera, A.

    2016-05-01

    We report on first experimental observations of nuclear fermionic and bosonic components displaying different behaviours in the decay of hot Ca projectile-like sources produced in mid-peripheral collisions at sub-Fermi energies. The experimental setup, constituted by the coupling of the INDRA 4π detector array to the forward angle VAMOS magnetic spectrometer, allowed to reconstruct the mass, charge and excitation energy of the decaying hot projectile-like sources. By means of quantum-fluctuation analysis techniques, temperatures and local partial densities of bosons and fermions could be correlated to the excitation energy of the reconstructed system. The results are consistent with the production of dilute mixed systems of bosons and fermions, where bosons experience higher phase-space and energy density as compared to the surrounding fermionic gas. Our findings recall phenomena observed in the study of Bose condensates and Fermi gases in atomic traps despite the different scales.

  12. HOT CELL BUILDING, TRA632, INTERIOR. CELL 3, "HEAVY" CELL. CAMERA ...

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

    HOT CELL BUILDING, TRA-632, INTERIOR. CELL 3, "HEAVY" CELL. CAMERA FACES WEST TOWARD BUILDING EXIT. OBSERVATION WINDOW AT LEFT EDGE OF VIEW. INL NEGATIVE NO. HD46-28-4. Mike Crane, Photographer, 2/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  13. Characterisation of hot particles remaining in soils from Palomares (Spain) using a nuclear microprobe

    NASA Astrophysics Data System (ADS)

    López, J. García; Jiménez-Ramos, M. C.; García-León, M.; García-Tenorio, R.

    2007-07-01

    More than 40 years ago, an aircraft accident took place in Palomares (Spain) which involved the destruction of two nuclear weapons. A portion of the remaining transuranic contamination in the affected soils is present in the form of small high activity concentration particles (10-100 μm), also called "hot particles", which contain plutonium and uranium. Several hot particles have been isolated and identified from the superficial soils recently collected in the zone affected by the accident. The isolation was carried out by screening the soil using gamma-ray spectrometry, through discrimination of the high activity concentrations of 241Am in the samples which indicates the presence of plutonium. The hot particles, composed of several elements with very high atomic number, could be easily identified by scanning electron microscopy (SEM) in backscattering electron image (BSE) mode. Moreover, their morphology and size were also studied using SEM in secondary electron (SE) mode. In this work, the hot particles have been investigated with the nuclear microprobe of the National Accelerator Centre (CNA) in Seville. Compositional analysis, mapping and depth distribution of different elements have been performed by a simultaneous combination of Particle Induced X-ray Emission (PIXE) and Rutherford Backscattering Spectrometry (RBS). Uranium and plutonium have been identified in the PIXE spectra as the main components of the particles, whereas the concentration of Americium is two orders of magnitude smaller. In addition, an estimation of the particles density has been obtained by comparison of the RBS results with the particles thickness directly determined by SEM.

  14. 324 and 325 Building hot cell cleanout program: Decontamination of C-Cell

    SciTech Connect

    Katayama, Y.B.; Holton, L.K. Jr.

    1989-10-01

    During FY 1989 the decontamination of C-Cell of Hanford's 324 Building was completed as part of the 324 and 325 Building Hot Cell Cleanout Program sponsored by the DOE Nuclear Energy's Surplus Facilities Management Program. The decontamination effort was completed using a series of remote and contact decontamination techniques. Initial radiation readings in C-Cell averaged 50 rad/hr and were reduced remotely to less than 200 mrad/hr using an alkaline foam cleaner followed by a 5000-psi water flush. Contact decontamination was then permissible using ultra high-pressure water, at 36,000 psi, further reducing the average radiation level in the cell to less than 86 mrem/hr. The approach used in decontaminating C-Cell resulted in a savings in radiation exposure of 87% and a cost savings of 39% compared to a hands-on procedure used in A-Cell, 324 Building in 1987. The radiation dose and the costs to achieve a 244-fold reduction in radiation contamination were 1.65 mrem per ft{sup 2} and $96 per ft{sup 2} of cell surface area. 14 figs., 4 tabs.

  15. Zirconium Recycle Test Equipment for Hot Cell Operations

    SciTech Connect

    Collins, Emory D.; DelCul, Guillermo Daniel; Spencer, Barry B.; Bradley, Eric Craig; Brunson, Ronald Ray

    2015-01-30

    The equipment components and assembly support work were modified for optimized, remote hot cell operations to complete this milestone. The modifications include installation of a charging door, Swagelok connector for the off-gas line between the reactor and condenser, and slide valve installation to permit attachment/replacement of the product salt collector bottle.

  16. Locating hot and cold-legs in a nuclear powered steam generation system

    DOEpatents

    Ekeroth, Douglas E.; Corletti, Michael M.

    1993-01-01

    A nuclear reactor steam generator includes a reactor vessel for heating water and a steam generator with a pump casing at the lowest point on the steam generator. A cold-leg pipe extends horizontally between the steam generator and the reactor vessel to return water from the steam generator to the reactor vessel. The bottom of the cold-leg pipe is at a first height above the bottom of the reactor vessel. A hot-leg pipe with one end connected to the steam generator and a second end connected to the reactor vessel has a first pipe region extending downwardly from the steam generator to a location between the steam generator and the reactor vessel at which a bottom of the hot-leg pipe is at a second height above the bottom of the reactor vessel. A second region extends from that location in a horizontal direction at the second height to the point at which the hot-leg pipe connects to the reactor vessel. A pump is attached to the casing at a location below the first and second heights and returns water from the steam generator to the reactor vessel over the cold-leg. The first height is greater than the second height and the bottom of the steam generator is at a height above the bottom of the reactor vessel that is greater than the first and second heights. A residual heat recovery pump is below the hot-leg and has an inlet line from the hot-leg that slopes down continuously to the pump inlet.

  17. Locating hot and cold-legs in a nuclear powered steam generation system

    DOEpatents

    Ekeroth, D.E.; Corletti, M.M.

    1993-11-16

    A nuclear reactor steam generator includes a reactor vessel for heating water and a steam generator with a pump casing at the lowest point on the steam generator. A cold-leg pipe extends horizontally between the steam generator and the reactor vessel to return water from the steam generator to the reactor vessel. The bottom of the cold-leg pipe is at a first height above the bottom of the reactor vessel. A hot-leg pipe with one end connected to the steam generator and a second end connected to the reactor vessel has a first pipe region extending downwardly from the steam generator to a location between the steam generator and the reactor vessel at which a bottom of the hot-leg pipe is at a second height above the bottom of the reactor vessel. A second region extends from that location in a horizontal direction at the second height to the point at which the hot-leg pipe connects to the reactor vessel. A pump is attached to the casing at a location below the first and second heights and returns water from the steam generator to the reactor vessel over the cold-leg. The first height is greater than the second height and the bottom of the steam generator is at a height above the bottom of the reactor vessel that is greater than the first and second heights. A residual heat recovery pump is below the hot-leg and has an inlet line from the hot-leg that slopes down continuously to the pump inlet. 2 figures.

  18. Arc-Heater Facility for Hot Hydrogen Exposure of Nuclear Thermal Rocket Materials

    NASA Technical Reports Server (NTRS)

    Litchford, Ron J.; Foote, John P.; Wang,Ten-See; Hickman, Robert; Panda, Binayak; Dobson, Chris; Osborne, Robin; Clifton, Scooter

    2006-01-01

    A hyper-thermal environment simulator is described for hot hydrogen exposure of nuclear thermal rocket material specimens and component development. This newly established testing capability uses a high-power, multi-gas, segmented arc-heater to produce high-temperature pressurized hydrogen flows representative of practical reactor core environments and is intended to serve. as a low cost test facility for the purpose of investigating and characterizing candidate fueUstructura1 materials and improving associated processing/fabrication techniques. Design and development efforts are thoroughly summarized, including thermal hydraulics analysis and simulation results, and facility operating characteristics are reported, as determined from a series of baseline performance mapping tests.

  19. Acoustic emission monitoring of hot functional testing: Watts Bar Unit 1 Nuclear Reactor

    SciTech Connect

    Hutton, P.H.; Dawson, J.F.; Friesel, M.A.; Harris, J.C.; Pappas, R.A.

    1984-06-01

    Acoustic emission (AE) monitoring of selected pressure boundary areas at TVA's Watts Bar, Unit 1 Nuclear Power Plant during hot functional preservice testing is described in this report. The report deals with background, methodology, and results. The work discussed here is a major milestone in a program supported by NRC to develop and demonstrate application of AE monitoring for continuous surveillance of reactor pressure boundaries to detect and evaluate growing flaws. The subject work demonstrated that anticipated problem areas can be overcome. Work is continuing toward AE monitoring during reactor operation.

  20. Phase transition of the baryon-antibaryon plasma in hot and dense nuclear matter

    NASA Astrophysics Data System (ADS)

    Lavagno, A.; Iazzi, F.; Pigato, D.

    2014-02-01

    We investigate the presence of thermodynamic instabilities in a hot and dense nuclear medium where a phase transition from a gas of massive hadrons to a nearly massless baryon, antibaryon plasma can take place. The analysis is performed by requiring the global conservation of baryon number and zero net strangeness in the framework of an effective relativistic mean field theory with the inclusion of the Δ(1232)-isobars, hyperons and the lightest pseudoscalar and vector meson degrees of freedom. Similarly to the low density nuclear liquid-gas phase transition, we show that such a phase transition is characterized by both mechanical instability (fluctuations on the baryon density) that by chemical- diffusive instability (fluctuations on the strangeness concentration). It turns out that, in this situation, phases with different values of antibaryon-baryon ratios and strangeness content may coexist.

  1. Testing hot cell shielding in the fuel conditioning facility.

    PubMed

    Courtney, J C; Klann, R T

    1997-01-01

    A comprehensive shield test program for a hot cell complex, the Fuel Conditioning Facility at Argonne National Laboratory, has been completed with minimum radiation exposure to participants. The recently modified shielding design for two hot cells and their associated transfer paths for irradiated materials was analyzed and tested for attenuating gamma rays from mixed fission product sources. Testing was accomplished using 0.37 TBq (10 Ci) and 518 TBq (14,000 Ci) 60Co sources. Of specific concern were radiation levels around wall penetrations and the interface between transport casks and the cell floor. Detailed measurements were made for surfaces that bound the hot cells, a transfer tunnel between the two cells, and storage pits that extend below the floors of both cells. In addition to surface measurements, dose equivalent rates in adjacent corridors were determined when the larger source was exposed. Results indicate that with some administrative controls, the facility shields are adequate to meet the design criterion that limits annual dose to less than 10 mSv (1 rem) for facility workers.

  2. HOT DIFFUSE EMISSION IN THE NUCLEAR STARBURST REGION OF NGC 2903

    SciTech Connect

    Yukita, Mihoko; Irwin, Jimmy A.; Swartz, Douglas A.; Tennant, Allyn F.; Soria, Roberto

    2012-10-20

    We present a deep Chandra observation of the central regions of the late-type barred spiral galaxy NGC 2903. The Chandra data reveal soft (kT{sub e} {approx} 0.2-0.5 keV) diffuse emission in the nuclear starburst region and extending {approx}2' ({approx}5 kpc) to the north and west of the nucleus. Much of this soft hot gas is likely to be from local active star-forming regions; however, besides the nuclear region, the morphology of hot gas does not strongly correlate with the bar or other known sites of active star formation. The central {approx}650 pc radius starburst zone exhibits much higher surface brightness diffuse emission than the surrounding regions and a harder spectral component in addition to a soft component similar to the surrounding zones. We interpret the hard component as also being of thermal origin with kT{sub e} {approx} 3.6 keV and to be directly associated with a wind fluid produced by supernovae and massive star winds similar to the hard diffuse emission seen in the starburst galaxy M82. The inferred terminal velocity for this hard component, {approx}1100 km s{sup -1}, exceeds the local galaxy escape velocity suggesting a potential outflow into the halo and possibly escape from the galaxy gravitational potential. Morphologically, the softer extended emission from nearby regions does not display an obvious outflow geometry. However, the column density through which the X-rays are transmitted is lower in the zone to the west of the nucleus compared to that from the east and the surface brightness is relatively higher suggesting some of the soft hot gas originates from above the disk: viewed directly from the western zone but through the intervening disk of the host galaxy along sight lines from the eastern zone. There are several point-like sources embedded in the strong diffuse nuclear emission zone. Their X-ray spectra show them to likely be compact binaries. None of these detected point sources are coincident with the mass center of the

  3. Nuclear reprogramming in mammalian somatic cell nuclear cloning

    PubMed Central

    Tamada, H.; Kikyo, N.

    2007-01-01

    Nuclear cloning is still a developing technique used to create genetically identical animals by somatic cell nuclear transfer into unfertilized eggs. Despite an intensive effort in a number of laboratories, the success rate of obtaining viable offspring from this technique remains less than 5%. In the past few years many investigators reported the reprogramming of specific nuclear activities in cloned animals, such as genome-wide gene expression patterns, DNA methylation, genetic imprinting, histone modifications and telomere length regulation. The results highlight the tremendous difficulty the clones face to reprogram the original differentiation status of the donor nuclei. Nevertheless, nuclei prepared from terminally differentiated lymphocytes can overcome this barrier and produce apparently normal mice. Study of this striking nuclear reprogramming activity should significantly contribute to our understanding of cell differentiation in more physiological settings. PMID:15237217

  4. Hot-carrier solar cell NEGF-based simulations

    NASA Astrophysics Data System (ADS)

    Cavassilas, Nicolas; Michelini, Fabienne; Bescond, Marc; Joie, Thibault

    2016-03-01

    Ultra thin absorbers for the hot carrier solar cell applications are promising. Indeed, in these ultimate absorbers electronphonon scattering are reduced and thickness can be lower than the electron mean-free-path. In this case carriers reach contact ballistically. However it is important that the contact permits to extract these carriers. This theoretical study is about the extraction of the photogenerated carriers and particularly the ballistic extraction without any scattering. We show that quantum interaction between the ultra-thin absorber and the contact can be used to enhance the extraction. Particularly, a contact composed of a quantum well into a double barrier permits to increase the current compared to a simple contact. This improvement is due to a quantum resonance. This result is interesting for the hot carrier solar cells but also for all the ultra-thin cells.

  5. Workshop on instrumentation and analyses for a nuclear fuel reprocessing hot pilot plant

    SciTech Connect

    Babcock, S.M.; Feldman, M.J.; Wymer, R.G.; Hoffman, D.

    1980-05-01

    In order to assist in the study of instrumentation and analytical needs for reprocessing plants, a workshop addressing these needs was held at Oak Ridge National Laboratory from May 5 to 7, 1980. The purpose of the workshop was to incorporate the knowledge of chemistry and of advanced measurement techniques held by the nuclear and radiochemical community into ideas for improved and new plant designs for both process control and inventory and safeguards measurements. The workshop was athended by experts in nuclear and radiochemistry, in fuel recycle plant design, and in instrumentation and analysis. ORNL was a particularly appropriate place to hold the workshop since the Consolidated Fuel Reprocessing Program (CFRP) is centered there. Requirements for safeguarding the special nuclear materials involved in reprocessing, and for their timely measurement within the process, within the reprocessing facility, and at the facility boundaries are being studied. Because these requirements are becoming more numerous and stringent, attention is also being paid to the analytical requirements for these special nuclear materials and to methods for measuring the physical parameters of the systems containing them. In order to provide a focus for the consideration of the workshop participants, the Hot Experimental Facility (HEF) being designed conceptually by the CFRP was used as a basis for consideration and discussions.

  6. Shell effects in hot nuclei and their influence on nuclear composition in supernova matter

    SciTech Connect

    Nishimura, Suguru; Takano, Masatoshi

    2014-05-02

    We calculate nuclear composition in supernova (SN) matter explicitly taking into account the temperature dependence of nuclear shell effects. The abundance of nuclei in SN matter is important in the dynamics of core-collapse supernovae and, in recently constructed equations of state (EOS) for SN matter, the composition of nuclei are calculated assuming nuclear statistical equilibrium wherein the nuclear internal free energies govern the composition. However, in these EOS, thermal effects on the shell energy are not explicitly taken into account. To address this shortfall, we calculate herein the shell energies of hot nuclei and examine their influence on the composition of SN matter. Following a simplified macroscopic-microscopic approach, we first calculate single-particle (SP) energies by using a spherical Woods-Saxon potential. Then we extract shell energies at finite temperatures using Strutinsky method with the Fermi distribution as the average occupation probability of the SP levels. The results show that at relatively low temperatures, shell effects are still important and magic nuclei are abundant. However, at temperatures above approximately 2 MeV, shell effects are almost negligible, and the mass fractions with shell energies including the thermal effect are close to those obtained from a simple liquid drop model at finite temperatures.

  7. Somatic cell nuclear transfer in zebrafish.

    PubMed

    Siripattarapravat, Kannika; Pinmee, Boonya; Venta, Patrick J; Chang, Chia-Cheng; Cibelli, Jose B

    2009-10-01

    We developed a method for somatic cell nuclear transfer in zebrafish using laser-ablated metaphase II eggs as recipients, the micropyle for transfer of the nucleus and an egg activation protocol after nuclear reconstruction. We produced clones from cells of both embryonic and adult origins, although the latter did not give rise to live adult clones.

  8. Moving Cell Boundaries Drive Nuclear Shaping during Cell Spreading

    PubMed Central

    Li, Yuan; Lovett, David; Zhang, Qiao; Neelam, Srujana; Kuchibhotla, Ram Anirudh; Zhu, Ruijun; Gundersen, Gregg G.; Lele, Tanmay P.; Dickinson, Richard B.

    2015-01-01

    The nucleus has a smooth, regular appearance in normal cells, and its shape is greatly altered in human pathologies. Yet, how the cell establishes nuclear shape is not well understood. We imaged the dynamics of nuclear shaping in NIH3T3 fibroblasts. Nuclei translated toward the substratum and began flattening during the early stages of cell spreading. Initially, nuclear height and width correlated with the degree of cell spreading, but over time, reached steady-state values even as the cell continued to spread. Actomyosin activity, actomyosin bundles, microtubules, and intermediate filaments, as well as the LINC complex, were all dispensable for nuclear flattening as long as the cell could spread. Inhibition of actin polymerization as well as myosin light chain kinase with the drug ML7 limited both the initial spreading of cells and flattening of nuclei, and for well-spread cells, inhibition of myosin-II ATPase with the drug blebbistatin decreased cell spreading with associated nuclear rounding. Together, these results show that cell spreading is necessary and sufficient to drive nuclear flattening under a wide range of conditions, including in the presence or absence of myosin activity. To explain this observation, we propose a computational model for nuclear and cell mechanics that shows how frictional transmission of stress from the moving cell boundaries to the nuclear surface shapes the nucleus during early cell spreading. Our results point to a surprisingly simple mechanical system in cells for establishing nuclear shapes. PMID:26287620

  9. Moving Cell Boundaries Drive Nuclear Shaping during Cell Spreading.

    PubMed

    Li, Yuan; Lovett, David; Zhang, Qiao; Neelam, Srujana; Kuchibhotla, Ram Anirudh; Zhu, Ruijun; Gundersen, Gregg G; Lele, Tanmay P; Dickinson, Richard B

    2015-08-18

    The nucleus has a smooth, regular appearance in normal cells, and its shape is greatly altered in human pathologies. Yet, how the cell establishes nuclear shape is not well understood. We imaged the dynamics of nuclear shaping in NIH3T3 fibroblasts. Nuclei translated toward the substratum and began flattening during the early stages of cell spreading. Initially, nuclear height and width correlated with the degree of cell spreading, but over time, reached steady-state values even as the cell continued to spread. Actomyosin activity, actomyosin bundles, microtubules, and intermediate filaments, as well as the LINC complex, were all dispensable for nuclear flattening as long as the cell could spread. Inhibition of actin polymerization as well as myosin light chain kinase with the drug ML7 limited both the initial spreading of cells and flattening of nuclei, and for well-spread cells, inhibition of myosin-II ATPase with the drug blebbistatin decreased cell spreading with associated nuclear rounding. Together, these results show that cell spreading is necessary and sufficient to drive nuclear flattening under a wide range of conditions, including in the presence or absence of myosin activity. To explain this observation, we propose a computational model for nuclear and cell mechanics that shows how frictional transmission of stress from the moving cell boundaries to the nuclear surface shapes the nucleus during early cell spreading. Our results point to a surprisingly simple mechanical system in cells for establishing nuclear shapes.

  10. Environmental Assessment for decontaminating and decommissioning the General Atomics Hot Cell Facility. Final [report

    SciTech Connect

    1995-08-01

    This EA evaluates the proposed action to decontaminate and decommission GA`s hot cell facility in northern San Diego, CA. This facility has been used for DOE and commercial nuclear R&D for > 30 years. About 30,000 cubic feet of decontamination debris and up to 50,000 cubic feet of contaminated soil are to be removed. Low-level radioactive waste would be shipped for disposal. It was determined that the proposal does not constitute a major federal action significantly affecting the human environment according to NEPA; therefore, a finding of no significant impact is made, and an environmental impact statement is not required.

  11. Cell Shape Dependent Regulation of Nuclear Morphology

    PubMed Central

    Chen, Bo; Co, Carlos; Ho, Chia-Chi

    2015-01-01

    Recent studies suggest that actin filaments are essential in how a cell controls its nuclear shape. However, little is known about the relative importance of membrane tension in determining nuclear morphology. In this study, we used adhesive micropatterned substrates to alter the cellular geometry (aspect ratio, size, and shape) that allowed direct membrane tension or without membrane lateral contact with the nucleus and investigate nuclear shape remodeling and orientation on a series of rectangular shapes. Here we showed that at low cell aspect ratios the orientation of the nucleus was regulated by actin filaments while cells with high aspect ratios can maintain nuclear shape and orientation even when actin polymerization was blocked. A model adenocarcinoma cell showed similar behavior in the regulation of nuclear shape in response to changes in cell shape but actin filaments were essential in maintaining cell shape. Our results highlight the two distinct mechanisms to regulate nuclear shape through cell shape control and the difference between fibroblasts and a model cancerous cell in cell adhesion and cell shape control. PMID:26210179

  12. Cell shape dependent regulation of nuclear morphology.

    PubMed

    Chen, Bo; Co, Carlos; Ho, Chia-Chi

    2015-10-01

    Recent studies suggest that actin filaments are essential in how a cell controls its nuclear shape. However, little is known about the relative importance of membrane tension in determining nuclear morphology. In this study, we used adhesive micropatterned substrates to alter the cellular geometry (aspect ratio, size, and shape) that allowed direct membrane tension or without membrane lateral contact with the nucleus and investigate nuclear shape remodeling and orientation on a series of rectangular shapes. Here we showed that at low cell aspect ratios the orientation of the nucleus was regulated by actin filaments while cells with high aspect ratios can maintain nuclear shape and orientation even when actin polymerization was blocked. A model adenocarcinoma cell showed similar behavior in the regulation of nuclear shape in response to changes in cell shape but actin filaments were essential in maintaining cell shape. Our results highlight the two distinct mechanisms to regulate nuclear shape through cell shape control and the difference between fibroblasts and a model cancerous cell in cell adhesion and cell shape control. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Nuclear size, nuclear pore number and cell cycle.

    PubMed

    Maeshima, Kazuhiro; Iino, Haruki; Hihara, Saera; Imamoto, Naoko

    2011-01-01

    In eukaryotic cells, the nucleus is a complex and sophisticated organelle containing genomic DNA and supports essential cellular activities. Its surface contains many nuclear pore complexes (NPCs), channels for macromolecular transport between the cytoplasm and nucleus. It has been observed that the nuclear volume and the number of NPCs almost doubles during interphase in dividing cells, but the coordination of these events with the cell cycle was poorly understood, particularly in mammalian cells. Recently, we demonstrated that cyclin-dependent protein kinases (Cdks) control interphase NPC formation in dividing human cells. Cdks drive the very early step of NPC formation because Cdk inhibition suppressed the generation of "nascent pores," which are considered to be immature NPCs, and disturbed expression and localization of some nucleoporins. Cdk inhibition did not affect nuclear volume, suggesting that these two processes have distinct regulatory mechanisms in the cell cycle. The details of our experimental systems and finding are discussed in more depth. With new findings recently reported, we also discuss possible molecular mechanisms of interphase NPC formation.

  14. Temperature and momentum dependence of single-particle properties in hot asymmetric nuclear matter

    SciTech Connect

    Moustakidis, Ch. C.

    2008-11-15

    We have studied the effects of momentum-dependent interactions on the single-particle properties of hot asymmetric nuclear matter. In particular, the single-particle potential of protons and neutrons as well as the symmetry potential have been studied within a self-consistent model using a momentum-dependent effective interaction. In addition, the isospin splitting of the effective mass has been derived from the above model. In each case temperature effects have been included and analyzed. The role of the specific parametrization of the effective interaction used in the present work has been investigated. It has been concluded that the behavior of the symmetry potential depends strongly on the parametrization of the interaction part of the energy density and the momentum dependence of the regulator function. The effects of the parametrization have been found to be less pronounced on the isospin mass splitting.

  15. Optical manipulation of hot nanoparticles can mediate selected cell fusion

    NASA Astrophysics Data System (ADS)

    Oddershede, Lene B.; Bahadori, Azra; Bendix, Poul M.

    2017-04-01

    Metallic nanoparticles with diameters from 10 nm to 250 nm can be optically trapped and manipulated in 3D using a single tightly focused near infrared laser beam. This will result in a significant heating of the particle and its vicinity, with temperature increases easily reaching hundreds degrees Celsius. If such a hot metallic nanoparticle is brought into the contact zone between two cells or vesicles, this local temperature increase can cause a total fusion of the selected cells or vesicles. Upon fusion, both the membrane and the cargos become completely mixed and we also show that the cells remain viable after fusion. The presented method has potential for single-cell targeted drug delivery and for the creation of hybrid cells.

  16. Cell fusion and nuclear fusion in plants.

    PubMed

    Maruyama, Daisuke; Ohtsu, Mina; Higashiyama, Tetsuya

    2016-12-01

    Eukaryotic cells are surrounded by a plasma membrane and have a large nucleus containing the genomic DNA, which is enclosed by a nuclear envelope consisting of the outer and inner nuclear membranes. Although these membranes maintain the identity of cells, they sometimes fuse to each other, such as to produce a zygote during sexual reproduction or to give rise to other characteristically polyploid tissues. Recent studies have demonstrated that the mechanisms of plasma membrane or nuclear membrane fusion in plants are shared to some extent with those of yeasts and animals, despite the unique features of plant cells including thick cell walls and intercellular connections. Here, we summarize the key factors in the fusion of these membranes during plant reproduction, and also focus on "non-gametic cell fusion," which was thought to be rare in plant tissue, in which each cell is separated by a cell wall. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Classification of hot particles from the Chernobyl accident and nuclear weapons detonations by non-destructive methods.

    PubMed

    Zheltonozhsky, V; Mück, K; Bondarkov, M

    2001-01-01

    Both after the Chernobyl accident and nuclear weapon detonations, agglomerates of radioactive material, so-called hot particles, were released or formed which show a behaviour in the environment quite different from the activity released in gaseous or aerosol form. The differences in their characteristic properties, in the radionuclide composition and the uranium and actinide contents are described in detail for these particles. While nuclear bomb hot particles (both from fission and fusion bombs) incorporate well detectable trace amounts of 60Co and 152Eu, these radionuclides are absent in Chernobyl hot particles. In contrast, Chernobyl hot particles contain 125Sb and 144Ce which are absent in atomic bomb HPs. Obvious differences are also observable between fusion and fission bombs' hot particles (significant differences in 152Eu/l55Eu, 154Eu/155Eu and 238Pu/239Pu ratios) which facilitate the identification of HPs of unknown provensence. The ratio of 239Pu/240Pu in Chernobyl hot particles could be determined by a non-destructive method at 1:1.5. A non-destructive method to determine the content of non-radioactive elements by Kalpha-emission measurements was developed by which inactive Zr, Nb, Fe and Ni could be verified in the particles.

  18. On the road toward a hot carrier solar cell

    NASA Astrophysics Data System (ADS)

    Taylor, P. C.; Fields, J. D.; Collins, R. T.

    2015-09-01

    We suggest a new paradigm for solar cells that uses a nanostructured crystalline collector (silicon) in an amorphous absorber matrix (hydrogenated amorphous silicon). Previously amorphous absorbers have received no serious consideration because of their low carrier mobilities. Specifically, we demonstrate that carriers generated in the amorphous region are transported out of this region before losing their energy to heat. This result establishes the possibility of using a wide range of nanostructured amorphous matrices to dramatically increase the efficiencies of solar cells. The use of an amorphous absorber provides a highly desirable and flexible approach to producing low-cost, hot carrier solar cells. Since amorphous materials can be grown over a much wider composition space than crystalline materials, this surprising result greatly broadens the absorbing materials that can be used to dramatically increase the efficiencies of solar cells.

  19. Decontamination of the Plum Brook Reactor Facility Hot Cells

    SciTech Connect

    Peecook, K.M.

    2008-07-01

    The NASA Plum Brook Reactor Facility decommissioning project recently completed a major milestone with the successful decontamination of seven hot cells. The cells included thick concrete walls and leaded glass windows, manipulator arms, inter cell dividing walls, and roof slabs. There was also a significant amount of embedded conduit and piping that had to be cleaned and surveyed. Prior to work starting evaluation studies were performed to determine whether it was more cost effective to do this work using a full up removal approach (rip and ship) or to decontaminate the cells to below required clean up levels, leaving the bulk of the material in place. This paper looks at that decision process, how it was implemented, and the results of that effort including the huge volume of material that can now be used as fill during site restoration rather than being disposed of as LLRW. (authors)

  20. Technical feasibility and economics of retrofitting an existing nuclear power plant to cogeneration for hot water district heating

    SciTech Connect

    Kolb, J.O.; Bauman, H.F.; Jones, P.D.

    1984-04-01

    This report gives the results of a study of the hypothetical conversion of the Prairie Island Nuclear Plant of the Northern States Power Company to cogeneration operation to supply a future hot water district heating system load in the Twin Cities of Minneapolis-St. Paul. The conceptual design of the nuclear turbine retrofitted for cogeneration and of a hot water transmission system has been performed, and the capital investment and annual owning and operating costs have been estimated for thermal energy capacities of 600 and 1200 MW(t). Unit costs of thermal energy (in mid-1982 dollars/million Btu) have been estimated for cogenerated hot water at the plant gate and also for the most economic transmission system from Prairie Island to the Twin Cities. The economic results from the analysis of the Prairie Island plant and transmission route have been generalized for other transmission distances in other locations.

  1. Reconstitution of Nuclear Import in Permeabilized Cells

    PubMed Central

    Cassany, Aurelia; Gerace, Larry

    2012-01-01

    The trafficking of protein and RNA cargoes between the cytoplasm and the nucleus of eukaryotic cells, which is a major pathway involved in cell regulation, is mediated by nuclear transport sequences in the cargoes and by shuttling transport factors. The latter include receptors (karyopherins) that recognize the cargoes and carry them across the nuclear pore complex (NPC), and the small GTPase Ran, which modulates karyopherin–cargo binding. Nuclear import can be studied in vitro using digitonin-permeabilized cells, which are depleted of shuttling transport factors. Nuclear import can be reconstituted in the permeabilized cells with exogenous cytosol or with purified recombinant transport factors, and can be quantified by light microscopy of fluorescently labeled cargoes or by immunofluorescence staining. Here we describe procedures for in vitro nuclear import in permeabilized mammalian cells, and for the preparation of recombinant transport factors (importin α, importin β, importin 7, transportin, Ran, NTF2) and other reagents commonly used in the assay. This assay provides means to characterize the molecular mechanisms of nuclear import and to study the import requirements of specific cargoes. PMID:18951186

  2. Reconstitution of nuclear import in permeabilized cells.

    PubMed

    Cassany, Aurelia; Gerace, Larry

    2009-01-01

    The trafficking of protein and RNA cargoes between the cytoplasm and the nucleus of eukaryotic cells, which is a major pathway involved in cell regulation, is mediated by nuclear transport sequences in the cargoes and by shuttling transport factors. The latter include receptors (karyopherins) that recognize the cargoes and carry them across the nuclear pore complex (NPC), and the small GTPase Ran, which modulates karyopherin-cargo binding. Nuclear import can be studied in vitro using digitonin-permeabilized cells, which are depleted of shuttling transport factors. Nuclear import can be reconstituted in the permeabilized cells with exogenous cytosol or with purified recombinant transport factors, and can be quantified by light microscopy of fluorescently labeled cargoes or by immunofluorescence staining. Here we describe procedures for in vitro nuclear import in permeabilized mammalian cells, and for the preparation of recombinant transport factors (importin alpha, importin beta, importin 7, transportin, Ran, NTF2) and other reagents commonly used in the assay. This assay provides means to characterize the molecular mechanisms of nuclear import and to study the import requirements of specific cargoes.

  3. Somatic Rearrangement in B Cells: It's (Mostly) Nuclear Physics.

    PubMed

    Aiden, Erez Lieberman; Casellas, Rafael

    2015-08-13

    We discuss how principles of nuclear architecture drive typical gene rearrangements in B lymphocytes, whereas translocation hot spots and recurrent lesions reflect the extent of AID-mediated DNA damage and selection.

  4. Nuclear binding of cell cycle-related proteins: cyclin A versus proliferating cell nuclear antigen (PCNA).

    PubMed

    Stivala, L A; Scovassi, A I; Bianchi, L; Prosperi, E

    1995-01-01

    We have investigated the cell cycle-dependent nuclear binding of cyclin A and of the proliferating cell nuclear antigen (PCNA) in asynchronously growing human fibroblasts. To this purpose, we have applied flow cytometry immunofluorescence, a powerful technique for elucidating the cell cycle phase during which the nuclear binding occurs. We have observed that, in striking contrast with the distribution of nuclear-bound PCNA which is restricted to S phase, the immunofluorescence signal of the nuclear-bound form of cyclin A is high in the G1 and G2 phases of the cell cycle. These results suggest the involvement of nuclear-bound cyclin A in the G1/S and G2/M phase transitions.

  5. Hot Cell Examination of Weapons-Grade MOX Fuel

    SciTech Connect

    Morris, Robert Noel; Bevard, Bruce Balkcom; McCoy, Kevin

    2010-01-01

    The U.S. Department of Energy has decided to dispose of a portion of the nation s surplus weapons-grade plutonium by reconstituting it into mixed oxide (MOX) fuel and irradiating it in commercial power reactors. Four lead assemblies were manufactured with weapons-grade MOX and irradiated to a maximum fuel rod burnup of 47.3 MWd/kg. As part of the fuel qualification process, five fuel rods with varying burnups and plutonium contents were selected from one of the assemblies and shipped to Oak Ridge National Laboratory for hot cell examination. This is the first hot cell examination of weapons-grade MOX fuel. The rods have been examined nondestructively with the ADEPT apparatus and are currently being destructively examined. Examinations completed to date include length measurements, visual examination, gamma scanning, profilometry, eddy-current testing, gas measurement and analysis, and optical metallography. Representative results of these examinations are reviewed and found to be consistent with predictions and with prior experience with reactor-grade MOX fuel. The results will be used to support licensing of weapons-grade MOX for batch use in commercial power reactors.

  6. Effect of a hot water extract of Chlorella vulgaris on proliferation of IEC-6 cells.

    PubMed

    Song, Seo-Hyeon; Kim, In-Hye; Nam, Taek-Jeong

    2012-05-01

    Chlorella vulgaris, a unicellular microalgae, exerts various biological effects; however their effect on proliferation signaling pathways in normal cells has not been studied. We investigated the effect of hot water extracts of Chlorella vulgaris (CVE) on cell proliferation and related signaling pathways in rat intestinal epithelial cells (IEC-6). CVE increased the expression of insulin-like growth factor-I receptor (IGF-IR) and the phosphorylation of focal adhesion kinase (FAK) and Src. In addition, CVE induced activation of the mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K)/Akt pathways. We verified the increased phosphorylation of extracellular-signal-related kinase (ERK) and Akt and the increased expression of the PI3K regulatory subunit p85. CVE also influenced the canonical Wnt pathway through increased expression of the nuclear β-catenin, cyclin D1. Tyr-397 of FAK mediates interactions with Src homology 2 (SH2) domains in a number of other signaling proteins, including PI3K, PLC-γ, Shc, Grb7, Src and Nck2. Because CVE induced FAK activation, FAK may affect the Wnt pathway. Addition of a FAK inhibitor decreased the expression of nuclear β-catenin, cyclin D1 and c-myc, and increased the expression of cytosolic β-catenin. We conclude that CVE stimulated proliferation of IEC-6 cells via the MAPK, PI3K/Akt and canonical Wnt pathways, and that this affected the canonical Wnt pathway.

  7. A micro hot test of the Chalmers-GANEX extraction system on used nuclear fuel

    SciTech Connect

    Bauhn, L.; Hedberg, M.; Aneheim, E.; Ekberg, C.; Loefstroem-Engdahl, E.; Skarnemark, G.

    2013-07-01

    In the present study, a 'micro hot test' has been performed using the Chalmers-GANEX (Group Actinide Extraction) system for partitioning of used nuclear fuel. The test included a pre-extraction step using N,N-di-2- ethylhexyl-butyramide (DEHBA) in n-octanol to remove the bulk part of the uranium. This pre-extraction was followed by a group extraction of actinides using the mixture of TBP and CyMe{sub 4}-BTBP in cyclohexanone as suggested in the Chalmers-GANEX process, and a three stage stripping of the extracted actinides. Distribution ratios for the extractions and stripping were determined based on a combination of γ- and α-spectrometry, as well as ICP-MS measurements. Successful extraction of uranium, plutonium and the minor actinides neptunium, americium and curium was achieved. However, measurements also indicated that co-extraction of europium occurs to some extent during the separation. These results were expected based on previous experiments using trace concentrations of actinides and lanthanides. Since this test was only performed in one stage with respect to the group actinide extraction, it is expected that multi stage tests will give even better results. (authors)

  8. Human somatic cell nuclear transfer and cloning.

    PubMed

    2012-10-01

    This document presents arguments that conclude that it is unethical to use somatic cell nuclear transfer (SCNT) for infertility treatment due to concerns about safety; the unknown impact of SCNT on children, families, and society; and the availability of other ethically acceptable means of assisted reproduction. This document replaces the ASRM Ethics Committee report titled, "Human somatic cell nuclear transfer (cloning)," last published in Fertil Steril 2000;74:873-6. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  9. Cold and hot nuclear matter effects on charmonium production in p+Pb collisions at LHC energy

    NASA Astrophysics Data System (ADS)

    Chen, Baoyi; Guo, Tiecheng; Liu, Yunpeng; Zhuang, Pengfei

    2017-02-01

    We study cold and hot nuclear matter effects on charmonium production in p+Pb collisions at √{sNN} = 5.02 TeV in a transport approach. At the forward rapidity, the cold medium effect on all the c c bar states and the hot medium effect on the excited c c bar states only can explain well the J / ψ and ψ‧ yield and transverse momentum distribution measured by the ALICE collaboration, and we predict a significantly larger ψ‧pT broadening in comparison with J / ψ. However, we can not reproduce the J / ψ and ψ‧ data at the backward rapidity with reasonable cold and hot medium effects.

  10. Remote real time x-ray examination of fuel elements in a hot cell environment

    SciTech Connect

    Yapuncich, F.L.

    1993-03-01

    This report discusses the Remote Real Time X-ray System which will allow for detailed examination of fuel elements. This task will be accomplished in a highly radioactive hot cell environment. Two remote handling systems win be utilized at the examination station. One handling system will transfer the fuel element to and from the shielded x-ray system. A second handling system will allow for vertical and rotational inspection of the fuel elements. The process win include removing a single nuclear fuel element from a element fabrication magazine(EFM), positioning the fuel element within the shielding envelope of the x-ray system and transferring the fuel element from the station manipulator to the x-ray system manipulator, performing the x-ray inspection, and then transferring the fuel element to either the element storage magazine(ESM) or a reject bin.

  11. Remote real time x-ray examination of fuel elements in a hot cell environment

    SciTech Connect

    Yapuncich, F.L.

    1993-01-01

    This report discusses the Remote Real Time X-ray System which will allow for detailed examination of fuel elements. This task will be accomplished in a highly radioactive hot cell environment. Two remote handling systems win be utilized at the examination station. One handling system will transfer the fuel element to and from the shielded x-ray system. A second handling system will allow for vertical and rotational inspection of the fuel elements. The process win include removing a single nuclear fuel element from a element fabrication magazine(EFM), positioning the fuel element within the shielding envelope of the x-ray system and transferring the fuel element from the station manipulator to the x-ray system manipulator, performing the x-ray inspection, and then transferring the fuel element to either the element storage magazine(ESM) or a reject bin.

  12. Hot wire deposited hydrogenated amorphous silicon solar cells

    SciTech Connect

    Mahan, A.H.; Iwaniczko, E.; Nelson, B.P.; Reedy, R.C. Jr.; Crandall, R.S.

    1996-05-01

    This paper details the results of a study in which low H content, high deposition rate hot wire (HW) deposited amorphous silicon (a-Si:H) has been incorporated into a substrate solar cell. The authors find that the treatment of the top surface of the HW i layer while it is being cooled from its high deposition temperature is crucial to device performance. They present data concerning these surface treatments, and correlate these treatments with Schottky device performance. The authors also present first generation HW n-i-p solar cell efficiency data, where a glow discharge (GD) {mu}c-Si(p) layer was added to complete the partial devices. No light trapping layer was used to increase the device Jsc. Their preliminary investigations have yielded efficiencies of up to 6.8% for a cell with a 4000 {Angstrom} thick HW i-layer, which degrade less than 10% after a 900 hour light soak. The authors suggest avenues for further improvement of their devices.

  13. Single cell elemental analysis using nuclear microscopy

    NASA Astrophysics Data System (ADS)

    Ren, M. Q.; Thong, P. S. P.; Kara, U.; Watt, F.

    1999-04-01

    The use of Particle Induced X-ray Emission (PIXE), Rutherford Backscattering Spectrometry (RBS) and Scanning Transmission Ion Microscopy (STIM) to provide quantitative elemental analysis of single cells is an area which has high potential, particularly when the trace elements such as Ca, Fe, Zn and Cu can be monitored. We describe the methodology of sample preparation for two cell types, the procedures of cell imaging using STIM, and the quantitative elemental analysis of single cells using RBS and PIXE. Recent work on single cells at the Nuclear Microscopy Research Centre,National University of Singapore has centred around two research areas: (a) Apoptosis (programmed cell death), which has been recently implicated in a wide range of pathological conditions such as cancer, Parkinson's disease etc, and (b) Malaria (infection of red blood cells by the malaria parasite). Firstly we present results on the elemental analysis of human Chang liver cells (ATTCC CCL 13) where vanadium ions were used to trigger apoptosis, and demonstrate that nuclear microscopy has the capability of monitoring vanadium loading within individual cells. Secondly we present the results of elemental changes taking place in individual mouse red blood cells which have been infected with the malaria parasite and treated with the anti-malaria drug Qinghaosu (QHS).

  14. Nuclear myosin I regulates cell membrane tension

    PubMed Central

    Venit, Tomáš; Kalendová, Alžběta; Petr, Martin; Dzijak, Rastislav; Pastorek, Lukáš; Rohožková, Jana; Malohlava, Jakub; Hozák, Pavel

    2016-01-01

    Plasma membrane tension is an important feature that determines the cell shape and influences processes such as cell motility, spreading, endocytosis and exocytosis. Unconventional class 1 myosins are potent regulators of plasma membrane tension because they physically link the plasma membrane with adjacent cytoskeleton. We identified nuclear myosin 1 (NM1) - a putative nuclear isoform of myosin 1c (Myo1c) - as a new player in the field. Although having specific nuclear functions, NM1 localizes predominantly to the plasma membrane. Deletion of NM1 causes more than a 50% increase in the elasticity of the plasma membrane around the actin cytoskeleton as measured by atomic force microscopy. This higher elasticity of NM1 knock-out cells leads to 25% higher resistance to short-term hypotonic environment and rapid cell swelling. In contrast, overexpression of NM1 in wild type cells leads to an additional 30% reduction of their survival. We have shown that NM1 has a direct functional role in the cytoplasm as a dynamic linker between the cell membrane and the underlying cytoskeleton, regulating the degree of effective plasma membrane tension. PMID:27480647

  15. Mycobacterium avium complex in day care hot water systems, and persistence of live cells and DNA in hot water pipes.

    PubMed

    Bukh, Annette S; Roslev, Peter

    2014-04-01

    The Mycobacterium avium complex (MAC) is a group of opportunistic human pathogens that may thrive in engineered water systems. MAC has been shown to occur in drinking water supplies based on surface water, but less is known about the occurrence and persistence of live cells and DNA in public hot water systems based on groundwater. In this study, we examined the occurrence of MAC in hot water systems of public day care centers and determined the persistence of live and dead M. avium cells and naked DNA in model systems with the modern plumbing material cross-linked polyethylene (PEX). The occurrence of MAC and co-occurrence of Legionella spp. and Legionella pneumophila were determined using cultivation and qPCR. Co-occurrences of MAC and Legionella were detected in water and/or biofilms in all hot water systems at temperatures between 40 and 54 °C. Moderate correlations were observed between abundance of culturable MAC and that of MAC genome copies, and between MAC and total eubacterial genome copies. No quantitative relationship was observed between occurrence of Legionella and that of MAC. Persistence in hot water of live and dead M. avium cells and naked DNA was studied using PEX laboratory model systems at 44 °C. Naked DNA and DNA in dead M. avium cells persisted for weeks. Live M. avium increased tenfold in water and biofilms on PEX. The results suggest that water and biofilms in groundwater-based hot water systems can constitute reservoirs of MAC, and that amplifiable naked DNA is relatively short-lived, whereas PEX plumbing material supports persistence and proliferation of M. avium.

  16. HOT CELL BUILDING, TRA632. OBLIQUE VIEW OF NORTH (LONG) AND ...

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

    HOT CELL BUILDING, TRA-632. OBLIQUE VIEW OF NORTH (LONG) AND WEST SIDES OF BUILDING. CHARGING PORT TO HOT CELL NO. 3 AND TEST TRAIN ASSEMBLY FACILITY AT RIGHT OF VIEW. CAMERA FACING SOUTHEAST. INL NEGATIVE NO. HD46-33-1. Mike Crane, Photographer, 4/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  17. 116. ARAI Details of hot cell section of building ARA626. ...

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

    116. ARA-I Details of hot cell section of building ARA-626. Shows manipulator openings in operating face of hot cell, start/stop buttons, and other details. Norman Engineering Company 961/area/SF-626-E-6. Date: January 1959. Ineel index code no. 068-0626-10-613-102731. - Idaho National Engineering Laboratory, Army Reactors Experimental Area, Scoville, Butte County, ID

  18. A&M. Hot cell annex (TAN633) interior of operating gallery. Camera ...

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

    A&M. Hot cell annex (TAN-633) interior of operating gallery. Camera probably facing south. At each side of the viewing windows are "master" manipulators which control "slaves" within hot cell. Date: March 2004. INEEL negative no. HD-39-2-3 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  19. Metal Hydrides as hot carrier cell absorber materials

    NASA Astrophysics Data System (ADS)

    Wang, Pei; Wen, Xiaoming; Shrestha, Santosh; Conibeer, Gavin; Aguey-Zinsou, Kondo-Francois

    2016-09-01

    The hot Carrier Solar Cell (HCSC) allows the photon-induced hot carriers (the carriers with energy larger than the band gap) to be collected before they completely thermalise. The absorber of the HCSC should have a large phononic band gap to supress Klemens Decay, which results in a slow carrier cooling speed. In fact, a large phononic band gap likely exists in a binary compound whose constituent elements have a large mass ratio between each other. Binary hydrides with their overwhelming mass ratio of the constituent elements are important absorber candidates. Study on different types of binary hydrides as potential absorber candidates is presented in this paper. Many binary transition metal hydrides have reported theoretical or experimental phonon dispersion charts which show large phononic band gaps. Among these hydrides, the titanium hydride (TiHX) is outstanding because of its low cost, easy fabrication process and is relatively inert to air and water. A TiHX thin film is fabricated by directly hydrogenating an evaporated titanium thin film. Characterisation shows good crystal quality and the hydrogenation process is believed to be successful. Ultrafast transient absorption (TA) spectroscopy is used to study the electron cooling time of TiHX. The result is very noisy due to the low absorption and transmission of the sample. The evolution of the TA curves has been explained by band to band transition using the calculated band structure of TiH2. Though not reliable due to the high noise, decay time fitting at 700nm and 600nm shows a considerably slow carrier cooling speed of the sample.

  20. Human embryonic stem cells derived by somatic cell nuclear transfer.

    PubMed

    Tachibana, Masahito; Amato, Paula; Sparman, Michelle; Gutierrez, Nuria Marti; Tippner-Hedges, Rebecca; Ma, Hong; Kang, Eunju; Fulati, Alimujiang; Lee, Hyo-Sang; Sritanaudomchai, Hathaitip; Masterson, Keith; Larson, Janine; Eaton, Deborah; Sadler-Fredd, Karen; Battaglia, David; Lee, David; Wu, Diana; Jensen, Jeffrey; Patton, Phillip; Gokhale, Sumita; Stouffer, Richard L; Wolf, Don; Mitalipov, Shoukhrat

    2013-06-06

    Reprogramming somatic cells into pluripotent embryonic stem cells (ESCs) by somatic cell nuclear transfer (SCNT) has been envisioned as an approach for generating patient-matched nuclear transfer (NT)-ESCs for studies of disease mechanisms and for developing specific therapies. Past attempts to produce human NT-ESCs have failed secondary to early embryonic arrest of SCNT embryos. Here, we identified premature exit from meiosis in human oocytes and suboptimal activation as key factors that are responsible for these outcomes. Optimized SCNT approaches designed to circumvent these limitations allowed derivation of human NT-ESCs. When applied to premium quality human oocytes, NT-ESC lines were derived from as few as two oocytes. NT-ESCs displayed normal diploid karyotypes and inherited their nuclear genome exclusively from parental somatic cells. Gene expression and differentiation profiles in human NT-ESCs were similar to embryo-derived ESCs, suggesting efficient reprogramming of somatic cells to a pluripotent state.

  1. Analytical Chemistry Laboratory (ACL) procedure compendium. Volume 7, Safety operation procedure for hot cell

    SciTech Connect

    Not Available

    1993-08-01

    This volume contains the interim change notice for the safety operation procedure for hot cell. It covers the master-slave manipulators, dry waste removal, cell transfers, hoists, cask handling, liquid waste system, and physical characterization of fluids.

  2. Minireview: Nuclear Receptors, Hematopoiesis, and Stem Cells

    PubMed Central

    Chute, John P.; Ross, Joel R.; McDonnell, Donald P.

    2010-01-01

    Nuclear receptors (NRs) regulate a panoply of biological processes, including the function and development of cells within the hematopoietic and immune system, such as erythrocytes, monocytes, and lymphocytes. Significantly less is known regarding the function of NRs in regulating the fate of hematopoietic stem cells (HSCs), the self-renewing, pluripotent cells that give rise to the entirety of the blood and immune systems throughout the lifetime of an individual. Several recent studies suggest, either directly or indirectly, a role for members of the NR family in regulating the differentiation and self-renewal of HSCs, embryonic stem cells, and induced pluripotent stem cells. Herein, we review in detail the function of specific NRs in controlling HSC and other stem cell fate and propose a framework through which these observations can be translated into therapeutic amplification of HSCs for clinical purposes. PMID:19934345

  3. Multiphysics Thermal-Fluid Design Analysis of a Non-Nuclear Tester for Hot-Hydrogen Materials and Component Development

    NASA Technical Reports Server (NTRS)

    Wang, Ten-See; Foote, John; Litchford, Ron

    2006-01-01

    The objective of this effort is to perform design analyses for a non-nuclear hot-hydrogen materials tester, as a first step towards developing efficient and accurate multiphysics, thermo-fluid computational methodology to predict environments for hypothetical solid-core, nuclear thermal engine thrust chamber design and analysis. The computational methodology is based on a multidimensional, finite-volume, turbulent, chemically reacting, thermally radiating, unstructured-grid, and pressure-based formulation. The multiphysics invoked in this study include hydrogen dissociation kinetics and thermodynamics, turbulent flow, convective, and thermal radiative heat transfers. The goals of the design analyses are to maintain maximum hot-hydrogen jet impingement energy and to minimize chamber wall heating. The results of analyses on three test fixture configurations and the rationale for final selection are presented. The interrogation of physics revealed that reactions of hydrogen dissociation and recombination are highly correlated with local temperature and are necessary for accurate prediction of the hot-hydrogen jet temperature.

  4. Multiphysics Thermal-Fluid Design Analysis of a Non-Nuclear Tester for Hot-Hydrogen Materials and Component Development

    SciTech Connect

    Wang, T.-S.; Foote, John; Litchford, Ron

    2006-01-20

    The objective of this effort is to perform design analyses for a non-nuclear hot-hydrogen materials tester, as a first step towards developing efficient and accurate multiphysics, thermo-fluid computational methodology to predict environments for hypothetical solid-core, nuclear thermal engine thrust chamber design and analysis. The computational methodology is based on a multidimensional, finite-volume, turbulent, chemically reacting, thermally radiating, unstructured-grid, and pressure-based formulation. The multiphysics invoked in this study include hydrogen dissociation kinetics and thermodynamics, turbulent flow, convective, and thermal radiative heat transfers. The goals of the design analyses are to maintain maximum hot-hydrogen jet impingement energy and to minimize chamber wall heating. The results of analyses on three test fixture configurations and the rationale for final selection are presented. The interrogation of physics revealed that reactions of hydrogen dissociation and recombination are highly correlated with local temperature and are necessary for accurate prediction of the hot-hydrogen jet temperature.

  5. Thermal Stress in HFEF Hot Cell Windows Due to an In-Cell Metal Fire

    DOE PAGES

    Solbrig, Charles W.; Warmann, Stephen A.

    2016-01-01

    This work investigates an accident during the pyrochemical extraction of Uranium and Plutonium from PWR spent fuel in an argon atmosphere hot cell. In the accident, the heavy metals (U and Pu) being extracted are accidentally exposed to air from a leaky instrument penetration which goes through the cell walls. The extracted pin size pieces of U and Pu metal readily burn when exposed to air. Technicians perform the electrochemical extraction using manipulators through a 4 foot thick hot cell concrete wall which protects them from the radioactivity of the spent fuel. Four foot thick windows placed in the wallmore » allow the technicians to visually control the manipulators. These windows would be exposed to the heat of the metal fire. As a result, this analysis determines if the thermal stress caused by the fire would crack the windows and if the heat would degrade the window seals allowing radioactivity to escape from the cell.« less

  6. Thermal Stress in HFEF Hot Cell Windows Due to an In-Cell Metal Fire

    SciTech Connect

    Solbrig, Charles W.; Warmann, Stephen A.

    2016-01-01

    This work investigates an accident during the pyrochemical extraction of Uranium and Plutonium from PWR spent fuel in an argon atmosphere hot cell. In the accident, the heavy metals (U and Pu) being extracted are accidentally exposed to air from a leaky instrument penetration which goes through the cell walls. The extracted pin size pieces of U and Pu metal readily burn when exposed to air. Technicians perform the electrochemical extraction using manipulators through a 4 foot thick hot cell concrete wall which protects them from the radioactivity of the spent fuel. Four foot thick windows placed in the wall allow the technicians to visually control the manipulators. These windows would be exposed to the heat of the metal fire. As a result, this analysis determines if the thermal stress caused by the fire would crack the windows and if the heat would degrade the window seals allowing radioactivity to escape from the cell.

  7. HOT CELL BUILDING, TRA632. EAST END OF BUILDING. CAMERA FACING ...

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

    HOT CELL BUILDING, TRA-632. EAST END OF BUILDING. CAMERA FACING WEST. TRUCK ENCLOSURE (1986) TO THE LEFT, SMALL ADDITION IN ITS SHADOW IS ENCLOSURE OVER METAL PORT INTO HOT CELL NO. 1 (THE OLDEST HOT CELL). NOTE PERSONNEL LADDER AND PLATFORM AT LOFT LEVEL USED WHEN SERVICING AIR FILTERS AND VENTS OF CELL NO. 1. INL NEGATIVE NO. HD46-32-4. Mike Crane, Photographer, 4/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  8. The Hanford spent nuclear metal fuel multi-canister overpack and vacuum drying {ampersand} hot conditioning process

    SciTech Connect

    Irwin, J.J.

    1996-05-15

    Nuclear production reactors operated at the U.S. Department of Energy`s Hanford Site from 1944 until 1988 to produce plutonium. Most of the irradiated fuel from these reactors was processed onsite to separate and recover the plutonium. When the processing facilities were closed in 1992, about 1,900 metric tons of unprocessed irradiated fuel remained in storage. Additional fuel was irradiated for research purposes or was shipped to the Hanford Site from offsite reactor facilities for storage or recovery of nuclear materials. The fuel inventory now in storage at the Hanford Site is predominantly N Reactor irradiated fuel, a metallic uranium alloy that is coextruded into zircaloy-2 cladding. The Spent Nuclear Fuel Project has rommitted to an accelerated schedule for removing spent nuclear fuel from the Hanford Site K Basins to a new interim storage facility in the 200 Area. Under the current proposed accelerated schedule, retrieval of spent nuclear fuel stored in the K East and West Basins must begin by December 1997 and be completed by December 1999. A key part of this action is retrieving fuel canisters from the water-filled K Basin storage pools and transferring them into multi@ister overpacks (MCOS) that will be used to handle and process the fuel, then store it after conditioning. The Westinghouse Hanford Company has developed an integrated process to deal with the K Basin spent fuel inventory. The process consists of cleaning the fuel, packaging it into MCOS, vacuum drying it at the K Basins, then transporting it to the Canister Storage Building (CSB) for staging, hot conditioning, and interim storage. This presentation dekribes the MCO function, design, and life-cycle, including an overview of the vacuum drying and hot conditioning processes.

  9. Solid oxide fuel cell systems with hot zones having improved reactant distribution

    DOEpatents

    Poshusta, Joseph C.; Booten, Charles W.; Martin, Jerry L.

    2012-11-06

    A Solid Oxide Fuel Cell (SOFC) system having a hot zone with a center cathode air feed tube for improved reactant distribution, a CPOX reactor attached at the anode feed end of the hot zone with a tail gas combustor at the opposing end for more uniform heat distribution, and a counter-flow heat exchanger for efficient heat retention.

  10. Solid oxide fuel cell systems with hot zones having improved reactant distribution

    DOEpatents

    Poshusta, Joseph C; Booten, Charles W; Martin, Jerry L

    2013-12-24

    A Solid Oxide Fuel Cell (SOFC) system having a hot zone with a center cathode air feed tube for improved reactant distribution, a CPOX reactor attached at the anode feed end of the hot zone with a tail gas combustor at the opposing end for more uniform heat distribution, and a counter-flow heat exchanger for efficient heat retention.

  11. Solid oxide fuel cell systems with hot zones having improved reactant distribution

    DOEpatents

    Poshusta, Joseph C.; Booten, Charles W.; Martin, Jerry L.

    2016-05-17

    A Solid Oxide Fuel Cell (SOFC) system having a hot zone with a center cathode air feed tube for improved reactant distribution, a CPOX reactor attached at the anode feed end of the hot zone with a tail gas combustor at the opposing end for more uniform heat distribution, and a counter-flow heat exchanger for efficient heat retention.

  12. Concentration-dependent Effects of Nuclear Lamins on Nuclear Size in Xenopus and Mammalian Cells*

    PubMed Central

    Jevtić, Predrag; Edens, Lisa J.; Li, Xiaoyang; Nguyen, Thang; Chen, Pan; Levy, Daniel L.

    2015-01-01

    A fundamental question in cell biology concerns the regulation of organelle size. While nuclear size is exquisitely controlled in different cell types, inappropriate nuclear enlargement is used to diagnose and stage cancer. Clarifying the functional significance of nuclear size necessitates an understanding of the mechanisms and proteins that control nuclear size. One structural component implicated in the regulation of nuclear morphology is the nuclear lamina, a meshwork of intermediate lamin filaments that lines the inner nuclear membrane. However, there has not been a systematic investigation of how the level and type of lamin expression influences nuclear size, in part due to difficulties in precisely controlling lamin expression levels in vivo. In this study, we circumvent this limitation by studying nuclei in Xenopus laevis egg and embryo extracts, open biochemical systems that allow for precise manipulation of lamin levels by the addition of recombinant proteins. We find that nuclear growth and size are sensitive to the levels of nuclear lamins, with low and high concentrations increasing and decreasing nuclear size, respectively. Interestingly, each type of lamin that we tested (lamins B1, B2, B3, and A) similarly affected nuclear size whether added alone or in combination, suggesting that total lamin concentration, and not lamin type, is more critical to determining nuclear size. Furthermore, we show that altering lamin levels in vivo, both in Xenopus embryos and mammalian tissue culture cells, also impacts nuclear size. These results have implications for normal development and carcinogenesis where both nuclear size and lamin expression levels change. PMID:26429910

  13. Hot compression process for making edge seals for fuel cells

    DOEpatents

    Dunyak, Thomas J.; Granata, Jr., Samuel J.

    1994-01-01

    A hot compression process for forming integral edge seals in anode and cade assemblies wherein the assemblies are made to a nominal size larger than a finished size, beads of AFLAS are applied to a band adjacent the peripheral margins on both sides of the assemblies, the assemblies are placed in a hot press and compressed for about five minutes with a force sufficient to permeate the peripheral margins with the AFLAS, cooled and cut to finished size.

  14. Post-irradiation-examination of irradiated fuel outside the hot cell

    SciTech Connect

    Dawn E. Janney; Adam B. Robinson; Thomas P. O'Holleran; R. Paul Lind; Marc Babcock; Laurence C. Brower; Julie Jacobs; Pamela K. Hoggan

    2007-09-01

    Because of their high radioactivity, irradiated fuels are commonly examined in a hot cell. However, the Idaho National Laboratory (INL) has recently investigated irradiated U-Mo-Al metallic fuel from the Reduced Enrichment for Research and Test Reactors (RERTR) project using a conventional unshielded scanning electron microscope outside a hot cell. This examination was possible because of a two-step sample-preparation approach in which a small volume of fuel was isolated in a hot cell and shielding was introduced during later stages of sample preparation. The resulting sample contained numerous sample-preparation artifacts but allowed analysis of microstructures from selected areas.

  15. Evaluation of a Mobile Hot Cell Technology for Processing Idaho National Laboratory Remote-Handled Wastes

    SciTech Connect

    B.J. Orchard; L.A. Harvego; R.P. Miklos; F. Yapuncich; L. Care

    2009-03-01

    The Idaho National Laboratory (INL) currently does not have the necessary capabilities to process all remote-handled wastes resulting from the Laboratory’s nuclear-related missions. Over the years, various U.S. Department of Energy (DOE)-sponsored programs undertaken at the INL have produced radioactive wastes and other materials that are categorized as remote-handled (contact radiological dose rate > 200 mR/hr). These materials include Spent Nuclear Fuel (SNF), transuranic (TRU) waste, waste requiring geological disposal, low-level waste (LLW), mixed waste (both radioactive and hazardous per the Resource Conservation and Recovery Act [RCRA]), and activated and/or radioactively-contaminated reactor components. The waste consists primarily of uranium, plutonium, other TRU isotopes, and shorter-lived isotopes such as cesium and cobalt with radiological dose rates up to 20,000 R/hr. The hazardous constituents in the waste consist primarily of reactive metals (i.e., sodium and sodium-potassium alloy [NaK]), which are reactive and ignitable per RCRA, making the waste difficult to handle and treat. A smaller portion of the waste is contaminated with other hazardous components (i.e., RCRA toxicity characteristic metals). Several analyses of alternatives to provide the required remote-handling and treatment capability to manage INL’s remote-handled waste have been conducted over the years and have included various options ranging from modification of existing hot cells to construction of new hot cells. Previous analyses have identified a mobile processing unit as an alternative for providing the required remote-handled waste processing capability; however, it was summarily dismissed as being a potentially viable alternative based on limitations of a specific design considered. In 2008 INL solicited expressions of interest from Vendors who could provide existing, demonstrated technology that could be applied to the retrieval, sorting, treatment (as required), and

  16. Nuclear microscopy of rat colon epithelial cells

    NASA Astrophysics Data System (ADS)

    Ren, M.; Rajendran, Reshmi; Ng, Mary; Udalagama, Chammika; Rodrigues, Anna E.; Watt, Frank; Jenner, Andrew Michael

    2011-10-01

    Using Nuclear microscopy, we have investigated iron distributions in the colons of Sprague Dawley rats, in order to elucidate heme uptake. Four groups of five Sprague Dawley rats (mean weight 180 g) were fed different purified diets containing either heme diet (2.5% w/w hemoglobin), high fat diet (HFD) (18% w/w fat, 1% w/w cholesterol), 'western' diet (combination of hemoglobin 2.5% and 18% fat, 1% cholesterol) or control diet (7% w/w fat). After 4 weeks, animals were sacrificed by exsanguination after anaesthesia. Thin sections of frozen colon tissue were taken, freeze dried and scanned using nuclear microscopy utilising the techniques PIXE, RBS and STIM. The new data acquisition system (IonDaq) developed in CIBA was used to obtain high resolution images and line scans were used to map the iron distributions across the colon boundaries. The nuclear microscope results indicate that when HFD is given in addition to heme, the iron content of the epithelial cells that line the colon decreases, and the zinc in the smooth muscle wall increases. This implies that the level of heme and fat in diet has an important role in colon health, possibly by influencing epithelial cells directly or changing luminal composition such as bacterial flora or levels of metabolites and cytotoxins.

  17. Robot Work Platform for Large Hot Cell Deactivation

    SciTech Connect

    BITTEN, E.J.

    2000-05-01

    The 324 Building, located at the Hanford Site near Richland, Washington, is being deactivated to meet state and federal cleanup commitments. The facility is currently in its third year of a nine-year project to complete deactivation and closure for long-term surveillance and maintenance. The 324 building contains large hot cells that were used for high-radiation, high-contamination chemical process development and demonstrations. A major obstacle for the 324 deactivation project is the inability to effectively perform deactivation tasks within highly radioactive, contaminated environments. Current strategies use inefficient, resource intensive technologies that significantly impact the cost and schedule for deactivation. To meet mandated cleanup commitments, there is a need to deploy rapid, more efficient remote/robot technologies to minimize worker exposure, accelerate work tasks, and eliminate the need for multiple specialized tool design and procurement efforts. This paper describes the functions and performance requirements for a crane-deployed remote/robot Work Platform possessing full access capabilities. The remote/robot Work Platform will deploy commercially available off-the-shelf tools and end effectors to support Project cleanup goals and reduce overall project risk and cost. The intent of this system is to maximize the use of off-the-shelf technologies that minimize additional new, unproven, or novel designs. This paper further describes procurement strategy, the selection process, the selected technology, and the current status of the procurement and lessons learned. Funding, in part, has been provided by the US Department of Energy, Office of Science and Technology, Deactivation and Decommissioning Focus Area.

  18. Single-nanowire, low-bandgap hot carrier solar cells with tunable open-circuit voltage.

    PubMed

    Limpert, Steven; Burke, Adam; Chen, I-Ju; Anttu, Nicklas; Lehmann, Sebastian; Fahlvik, Sofia; Bremner, Stephen; Conibeer, Gavin J; Thelander, Claes; Pistol, Mats-Erik; Linke, Heiner

    2017-08-31

    Compared to traditional pn-junction photovoltaics, hot carrier solar cells offer potentially higher efficiency by extracting work from the kinetic energy of photogenerated "hot carriers" before they cool to the lattice temperature. Hot carrier solar cells have been demonstrated in high-bandgap ferroelectric insulators and GaAs/AlGaAs heterostructures, but so far not in low-bandgap materials, where the potential efficiency gain is highest. Recently, a high open-circuit voltage was demonstrated in an illuminated wurtzite InAs nanowire with a low bandgap of 0.39 eV, and was interpreted in terms of a photothermoelectric effect. Here, we point out that this device is a hot carrier solar cell and discuss its performance in those terms. In the demonstrated devices, InP heterostructures are used as energy filters in order to thermoelectrically harvest the energy of hot electrons photogenerated in InAs absorber segments. The obtained photovoltage depends on the heterostructure design of the energy filter and is therefore tunable. By using a high-resistance, thermionic barrier, an open-circuit voltage is obtained that is in excess of the Shockley-Queisser limit. These results provide generalizable insight into how to realize high voltage hot carrier solar cells in low-bandgap materials, and therefore are a step towards the demonstration of higher efficiency hot carrier solar cells. © 2017 IOP Publishing Ltd.

  19. Radiophase development in hot-pressed alkoxide-derived titanate ceramics for nuclear waste stabilization

    SciTech Connect

    Dickson, F.J. ); Mitamura, H. ); White, T.J. )

    1989-06-01

    This paper reports phase development as a function of hot-pressing temperature studied in alkoxide-derived titanate-based ceramics doped with a 10 wt% loading of a sodium-rich (NAR) and a sodium-poor (NAP) simulated high-level waste. Pyrochlore was found to be the most abundant phase in both calcine powders. A pseudobrookite phase existed metastably at hot-pressing temperatures between 890{degrees} and 920{degrees} C. After hot-pressing at 1100{degrees} C, the final phase assemblage for the NAP material consisted of zirconolite, hollandite-type, perovskite, alloy, and reduced rutile (Magneli phases). In addition, NAR samples contained hibonite, freudenbergite, and loveringite. Phase development was driven to completion over a very narrow temperature range ({le}50{degrees} C), beginning at 870{degrees} and 850{degrees} C for NAP and NAR, respectively, although full densification was not achieved below 1100{degrees} C. Both waste forms exhibited comparable microstructure and aqueous durability.

  20. Dirac-Hartree-Bogoliubov calculation for spherical and deformed hot nuclei: Temperature dependence of the pairing energy and gaps, nuclear deformation, nuclear radii, excitation energy, and entropy

    NASA Astrophysics Data System (ADS)

    Lisboa, R.; Malheiro, M.; Carlson, B. V.

    2016-02-01

    Background: Unbound single-particle states become important in determining the properties of a hot nucleus as its temperature increases. We present relativistic mean field (RMF) for hot nuclei considering not only the self-consistent temperature and density dependence of the self-consistent relativistic mean fields but also the vapor phase that takes into account the unbound nucleon states. Purpose: The temperature dependence of the pairing gaps, nuclear deformation, radii, binding energies, entropy, and caloric curves of spherical and deformed nuclei are obtained in self-consistent RMF calculations up to the limit of existence of the nucleus. Method: We perform Dirac-Hartree-Bogoliubov (DHB) calculations for hot nuclei using a zero-range approximation to the relativistic pairing interaction to calculate proton-proton and neutron-neutron pairing energies and gaps. A vapor subtraction procedure is used to account for unbound states and to remove long range Coulomb repulsion between the hot nucleus and the gas as well as the contribution of the external nucleon gas. Results: We show that p -p and n -n pairing gaps in the S10 channel vanish for low critical temperatures in the range Tcp≈0.6 -1.1 MeV for spherical nuclei such as 90Zr, 124Sn, and 140Ce and for both deformed nuclei 150Sm and 168Er. We found that superconducting phase transition occurs at Tcp=1.03 Δp p(0 ) for 90Zr, Tcp=1.16 Δp p(0 ) for 140Ce, Tcp=0.92 Δp p(0 ) for 150Sm, and Tcp=0.97 Δp p(0 ) for 168Er. The superfluidity phase transition occurs at Tcp=0.72 Δn n(0 ) for 124Sn, Tcp=1.22 Δn n(0 ) for 150Sm, and Tcp=1.13 Δn n(0 ) for 168Er. Thus, the nuclear superfluidity phase—at least for this channel—can only survive at very low nuclear temperatures and this phase transition (when the neutron gap vanishes) always occurs before the superconducting one, where the proton gap is zero. For deformed nuclei the nuclear deformation disappear at temperatures of about Tcs=2.0 -4.0 MeV , well above the

  1. Remote System Technologies for Deactivating Hanford Hot Cells (for WM'03 - abstract included)

    SciTech Connect

    BERLIN, G.T.

    2003-01-28

    Remote system technologies are being deployed by Fluor Hanford to help accelerate the deactivation of highly-radioactive hot cell facilities. This paper highlights the application of several remotely deployed technologies enabling the deactivation tasks.

  2. Nuclear Nonhistone Proteins in Murine Melanoma Cells

    PubMed Central

    Wikswo, Muriel A.; Mcguire, Joseph S.; Shansky, Janet E.; Boshes, Roger A.

    1976-01-01

    Nuclear nonhistone proteins (NHP's) have been implicated as regulatory agents involved in controlling genetic expression. Utilizing murine melanoma cells, we describe a method for isolating and fractionating NHP's which greatly increases the yield of these proteins as well as the level of resolution required for detecting small differences in particular NHP's. Mouse melanoma cells were grown in medium labeled with [3H]leucine. Following 48 hr of incubation, the cells were harvested and nuclei isolated. The NHP's were extracted from the nuclei in a series of steps which yielded four major fractions: NHP1, NHP2, NHP3, NHP4. This method solubilized 80-90% of the protein from the nuclear homogenate. The NHP fractions were then separated on DEAE-cellulose columns in a series of salt steps increasing in concentration from 0.05 to 0.50 M NaCl, followed by steps of 2 M NaCl and 4 and 7 M guanidine-hydrochloride. The 40 NHP fractions eluted from these columns were further separated on polyacrylamide-SDS gels and ranged in molecular weight from 9000 to 110,000 daltons. Differences were observed in the electrophoretic pattern of each of these 40 fractions. The high resolution of these fractionation procedures greatly enhances the possibility of observing small changes in proteins which may play a role in gene regulation. ImagesFIG. 2FIG. 5 PMID:997593

  3. Thermodynamics of Hot Nuclear Matter: 1978 in the Statistical Bootstrap Model

    NASA Astrophysics Data System (ADS)

    Rafelski, Johann; Hagedorn, Rolf

    We formulate the statistical bootstrap model for nuclear matter, and study its resulting thermodynamic properties at nuclear densities below the saturation density. We discuss the relevance of limiting temperature and the phase transition gas-`liquid' when the volume of the fireball grows with its energy.

  4. HOT CELL BUILDING, TRA632, INTERIOR. OPEN CORRIDOR ALONG SOUTH WALL ...

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

    HOT CELL BUILDING, TRA-632, INTERIOR. OPEN CORRIDOR ALONG SOUTH WALL OF BUILDING. CAMERA IS NEAR HOT CELL NO. 1, FACES WEST TOWARDS WALL OF TEST-TRAIN ASSEMBLY (TRA-632A). NOTE MOTORIZED RAIL CRANE ABOVE STAIRWAY. INL NEGATIVE NO. HD46-29-3. Mike Crane, Photographer, 2/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  5. HOT CELL BUILDING, TRA632. WEST END. CAMERA FACING EAST. ENCLOSURE ...

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

    HOT CELL BUILDING, TRA-632. WEST END. CAMERA FACING EAST. ENCLOSURE TO CHARGING PORT INTO HOT CELL NO. 3 IS ON LEFT SIDE, BUILT IN 1965. ON ITS RIGHT IS TEST TRAIN ASSEMBLY FACILITY, BUILT IN 1968. INL NEGATIVE NO. HD46-33-2. Mike Crane, Photographer, 4/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  6. DQO Summary Report for 324 and 327 Building Hot Cells D4 Project Waste Characterization

    SciTech Connect

    T.A. Lee

    2006-02-06

    This data quality objective (DQO) summary report provides the results of the DQO process conducted for waste characterization activities for the 324 and 327 Building hot cells decommission, deactivate, decontaminate, and demolish activities. This DQO summary report addresses the systems and processes related to the hot cells, air locks, vaults, tanks, piping, basins, air plenums, air ducts, filters, an adjacent elements that have high dose rates, high contamination levels, and/or suspect transuranic waste, which will require nonstandard D4 techniques.

  7. HOT CELL BUILDING, TRA632. WHILE STEEL BEAMS DEFINE FUTURE WALLS ...

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

    HOT CELL BUILDING, TRA-632. WHILE STEEL BEAMS DEFINE FUTURE WALLS OF THE BUILDING, SHEET STEEL DEFINES THE HOT CELL "BOX" ITSELF. THREE OPERATING WINDOWS ON LEFT; ONE VIEWING WINDOW ON RIGHT. TUBES WILL CONTAIN SERVICE AND CONTROL LEADS. SPACE BETWEEN INNER AND OUTER BOX WALLS WILL BE FILLED WITH SHIELDED WINDOWS AND BARETES CONCRETE. CAMERA FACES SOUTHEAST. INL NEGATIVE NO. 7933. Unknown Photographer, ca. 5/1953 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  8. A&M. Hot cell addition (TAN633). Floor plan, elevations. Arrangement of ...

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

    A&M. Hot cell addition (TAN-633). Floor plan, elevations. Arrangement of monorail along corridor, four hot cells, plug access openings, viewing windows, photo darkroom. Ralph M. Parsons 1229-13-ANP/GE-3-633-A-1. Date: December 1956 as redrawn in August 1998. Approved by INEEL Classification Office for public release. INEEL index code no. 034-0633-00-693-107315 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  9. 113. ARAI Hot cell (ARA626) Building wall sections and details ...

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

    113. ARA-I Hot cell (ARA-626) Building wall sections and details of radio chemistry lab. Shows high-bay roof over hot cells and isolation rooms below grade storage pit for fuel elements. Norman Engineering Company: 961-area/SF-626-A-4. Date: January 1959. Ineel index code no. 068-0626-00-613-102724. - Idaho National Engineering Laboratory, Army Reactors Experimental Area, Scoville, Butte County, ID

  10. Effect of donor cell type on nuclear remodelling in rabbit somatic cell nuclear transfer embryos.

    PubMed

    Tian, J; Song, J; Li, H; Yang, D; Li, X; Ouyang, H; Lai, L

    2012-08-01

    Cloned rabbits have been produced for many years by somatic cell nuclear transfer (SCNT). The efficiency of cloning by SCNT, however, has remained extremely low. Most cloned embryos degenerate in utero, and the few that develop to term show a high incidence of post-natal death and abnormalities. The cell type used for donor nuclei is an important factor in nuclear transfer (NT). As reported previously, NT embryos reconstructed with fresh cumulus cells (CC-embryos) have better developmental potential than those reconstructed with foetal fibroblasts (FF-embryos) in vivo and in vitro. The reason for this disparity in developmental capacity is still unknown. In this study, we compared active demethylation levels and morphological changes between the nuclei of CC-embryos and FF-embryos shortly after activation. Anti-5-methylcytosine immunofluorescence of in vivo-fertilized and cloned rabbit embryos revealed that there was no detectable active demethylation in rabbit zygotes or NT-embryos derived from either fibroblasts or CC. In the process of nuclear remodelling, however, the proportion of nuclei with abnormal appearance in FF-embryos was significantly higher than that in CC-embryos during the first cell cycle. Our study demonstrates that the nuclear remodelling abnormality of cloned rabbit embryos may be one important factor for the disparity in developmental success between CC-embryos and FF-embryos.

  11. Live-Cell Pyrophosphate Imaging by in Situ Hot-Spot Generation.

    PubMed

    Li, Mingmin; Li, Jin; Di, Huixia; Liu, Huiqiao; Liu, Dingbin

    2017-03-21

    Controlling the electromagnetic hot-spot generation is essential for surface-enhanced Raman scattering (SERS) assays. Current hot-spot-based SERS assays have been extensively studied in solutions or on substrates. However, probing biospecies by controlling the hot-spot assembly in living systems has not been demonstrated thus far. Herein, we report a background-free SERS probe for imaging pyrophosphate (PPi), a biochemically significant anion, in living cells. Intracellular PPi is able to induce the nanoparticle dimerization, thus creating an intense electromagnetic hot spot and dramatically enhancing the signal of the Raman reporters residing in the hot spot. More impressively, the reporter we used in this study provides a strong and sharp single peak in the cellular Raman-silent region (1800-2800 cm(-1)), thus eliminating the possible background interference. This strategy could be readily extended to detect other biomarkers by only replacing the recognition ligands.

  12. Hot-wall corrosion testing of simulated high level nuclear waste

    SciTech Connect

    Chandler, G.T.; Zapp, P.E.; Mickalonis, J.I.

    1995-01-01

    Three materials of construction for steam tubes used in the evaporation of high level radioactive waste were tested under heat flux conditions, referred to as hot-wall tests. The materials were type 304L stainless steel alloy C276, and alloy G3. Non-radioactive acidic and alkaline salt solutions containing halides and mercury simulated different high level waste solutions stored or processed at the United States Department of Energy`s Savannah River Site. Alloy C276 was also tested for corrosion susceptibility under steady-state conditions. The nickel-based alloys C276 and G3 exhibited excellent corrosion resistance under the conditions studied. Alloy C276 was not susceptible to localized corrosion and had a corrosion rate of 0.01 mpy (0.25 {mu}m/y) when exposed to acidic waste sludge and precipitate slurry at a hot-wall temperature of 150{degrees}C. Type 304L was susceptible to localized corrosion under the same conditions. Alloy G3 had a corrosion rate of 0.1 mpy (2.5 {mu}m/y) when exposed to caustic high level waste evaporator solution at a hot-wall temperature of 220{degrees}C compared to 1.1 mpy (28.0 {mu}/y) for type 304L. Under extreme caustic conditions (45 weight percent sodium hydroxide) G3 had a corrosion rate of 0.1 mpy (2.5 {mu}m/y) at a hot-wall temperature of 180{degrees}C while type 304L had a high corrosion rate of 69.4 mpy (1.8 mm/y).

  13. Actomyosin drives cancer cell nuclear dysmorphia and threatens genome stability

    PubMed Central

    Takaki, Tohru; Montagner, Marco; Serres, Murielle P.; Le Berre, Maël; Russell, Matt; Collinson, Lucy; Szuhai, Karoly; Howell, Michael; Boulton, Simon J.; Sahai, Erik; Petronczki, Mark

    2017-01-01

    Altered nuclear shape is a defining feature of cancer cells. The mechanisms underlying nuclear dysmorphia in cancer remain poorly understood. Here we identify PPP1R12A and PPP1CB, two subunits of the myosin phosphatase complex that antagonizes actomyosin contractility, as proteins safeguarding nuclear integrity. Loss of PPP1R12A or PPP1CB causes nuclear fragmentation, nuclear envelope rupture, nuclear compartment breakdown and genome instability. Pharmacological or genetic inhibition of actomyosin contractility restores nuclear architecture and genome integrity in cells lacking PPP1R12A or PPP1CB. We detect actin filaments at nuclear envelope rupture sites and define the Rho-ROCK pathway as the driver of nuclear damage. Lamin A protects nuclei from the impact of actomyosin activity. Blocking contractility increases nuclear circularity in cultured cancer cells and suppresses deformations of xenograft nuclei in vivo. We conclude that actomyosin contractility is a major determinant of nuclear shape and that unrestrained contractility causes nuclear dysmorphia, nuclear envelope rupture and genome instability. PMID:28737169

  14. Somatic Cell Nuclear Transfer in the Mouse

    NASA Astrophysics Data System (ADS)

    Kishigami, Satoshi; Wakayama, Teruhiko

    Somatic cell nuclear transfer (SCNT) has become a unique and powerful tool for epigenetic reprogramming research and gene manipulation in animals since “Dolly,” the first animal cloned from an adult cell was reported in 1997. Although the success rates of somatic cloning have been inefficient and the mechanism of reprogramming is still largely unknown, this technique has been proven to work in more than 10 mammalian species. Among them, the mouse provides the best model for both basic and applied research of somatic cloning because of its abounding genetic resources, rapid sexual maturity and propagation, minimal requirements for housing, etc. This chapter describes a basic protocol for mouse cloning using cumulus cells, the most popular cell type for NT, in which donor nuclei are directly injected into the oocyte using a piezo-actuated micromanipulator. In particular, we focus on a new, more efficient mouse cloning protocol using trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, which increases both in vitro and in vivo developmental rates from twofold to fivefold. This new method including TSA will be helpful to establish mouse cloning in many laboratories.

  15. Assembly dynamics of PML nuclear bodies in living cells.

    PubMed

    Brand, Peter; Lenser, Thorsten; Hemmerich, Peter

    2010-03-05

    The mammalian cell nucleus contains a variety of organelles or nuclear bodies which contribute to key nuclear functions. Promyelocytic leukemia nuclear bodies (PML NBs) are involved in the regulation of apoptosis, antiviral responses, the DNA damage response and chromatin structure, but their precise biochemical function in these nuclear pathways is unknown. One strategy to tackle this problem is to assess the biophysical properties of the component parts of these macromolecular assemblies in living cells. In this study we determined PML NB assembly dynamics by live cell imaging, combined with mathematical modeling. For the first time, dynamics of PML body formation were measured in cells lacking endogenous PML. We show that all six human nuclear PML isoforms are able to form nuclear bodies in PML negative cells. All isoforms exhibit individual exchange rates at NBs in PML positive cells but PML I, II, III and IV are static at nuclear bodies in PML negative cells, suggesting that these isoforms require additional protein partners for efficient exchange. PML V turns over at PML Nbs very slowly supporting the idea of a structural function for this isoform. We also demonstrate that SUMOylation of PML at Lysine positions K160 and/or K490 are required for nuclear body formation in vivo.We propose a model in which the isoform specific residence times of PML provide both, structural stability to function as a scaffold and flexibility to attract specific nuclear proteins for efficient biochemical reactions at the surface of nuclear bodies.MCS code: 92C37.

  16. T Helper Cell Tolerance to Ubiquitous Nuclear Antigens

    PubMed Central

    NAKKEN, BRITT; DAVIS, KAREN E.; PAN, ZIJIAN; BACHMANN, MICHAEL; FARRIS, A. DARISE

    2007-01-01

    Systemic autoimmune diseases are characterized by the development of anti-nuclear autoantibodies. In order to understand the immunologic events leading to the development of such antibodies, knowledge of mechanisms of immune tolerance to nuclear antigens is required. By utilizing adoptive T cell transfer strategies with transgenic mouse models expressing nuclear neo-self antigens, T cell tolerance to the lupus-related nuclear antigens human La and nRNP A has been demonstrated. These findings also indicate the existence in normal animals of autoreactive B cells continuously presenting nuclear antigen, suggesting that nuclear antigens are not sequestered from the immune system. Investigations of CD4+ T cell tolerance to non-nuclear antigens have revealed a number of mechanisms that protect the host from autoreactivity, including autoreactive T cell deletion, regulatory T cell development and anergy induction. Recent studies using T cell receptor and neo-self nuclear antigen transgenic mice are revealing the importance of such mechanisms in maintaining tolerance to nuclear antigens. Mechanisms of tolerogenic antigen presentation, identification of tolerogenic antigen source(s), and the pathways leading to loss of tolerance to nuclear antigens in systemic autoimmune disease states are currently being sought. PMID:14629620

  17. Charge transfer state versus hot exciton dissociation in polymer-fullerene blended solar cells.

    PubMed

    Lee, Jiye; Vandewal, Koen; Yost, Shane R; Bahlke, Matthias E; Goris, Ludwig; Baldo, Marc A; Manca, Jean V; Van Voorhis, Troy

    2010-09-01

    We examine the significance of hot exciton dissociation in two archetypical polymer-fullerene blend solar cells. Rather than evolving through a bound charge transfer state, hot processes are proposed to convert excitons directly into free charges. But we find that the internal quantum yields of carrier photogeneration are similar for both excitons and direct excitation of charge transfer states. The internal quantum yield, together with the temperature dependence of the current-voltage characteristics, is consistent with negligible impact from hot exciton dissociation.

  18. Submerged RadBall® deployments in Hanford Site hot cells containing 137CsCl capsules.

    PubMed

    Farfán, Eduardo B; Coleman, J Rusty; Stanley, Steven; Adamovics, John; Oldham, Mark; Thomas, Andrew

    2012-07-01

    The overall objective of this study was to demonstrate that a new technology, known as RadBall®, could locate submerged radiological hazards. RadBall® is a novel, passive, radiation detection device that provides a 3-D visualization of radiation from areas where measurements have not been previously possible due to lack of access or extremely high radiation doses. This technology has been under development during recent years, and all of its previous tests have included dry deployments. This study involved, for the first time, underwater RadBall® deployments in hot cells containing 137CsCl capsules at the U.S. Department of Energy's Hanford Site. RadBall® can be used to characterize a contaminated room, hot cell, or glovebox by providing the locations of the radiation sources and hazards, identifying the radionuclides present within the cell, and determining the radiation sources' strength (e.g., intensities or dose rates). These parameters have been previously determined for dry deployments; however, only the location of radiation sources and hazards can be determined for an underwater RadBall® deployment. The results from this study include 3-D images representing the location of the radiation sources within the Hanford Site cells. Due to RadBall®'s unique deployability and non-electrical nature, this technology shows significant promise for future characterization of radiation hazards prior to and during the decommissioning of contaminated nuclear facilities.

  19. Concentration-dependent Effects of Nuclear Lamins on Nuclear Size in Xenopus and Mammalian Cells.

    PubMed

    Jevtić, Predrag; Edens, Lisa J; Li, Xiaoyang; Nguyen, Thang; Chen, Pan; Levy, Daniel L

    2015-11-13

    A fundamental question in cell biology concerns the regulation of organelle size. While nuclear size is exquisitely controlled in different cell types, inappropriate nuclear enlargement is used to diagnose and stage cancer. Clarifying the functional significance of nuclear size necessitates an understanding of the mechanisms and proteins that control nuclear size. One structural component implicated in the regulation of nuclear morphology is the nuclear lamina, a meshwork of intermediate lamin filaments that lines the inner nuclear membrane. However, there has not been a systematic investigation of how the level and type of lamin expression influences nuclear size, in part due to difficulties in precisely controlling lamin expression levels in vivo. In this study, we circumvent this limitation by studying nuclei in Xenopus laevis egg and embryo extracts, open biochemical systems that allow for precise manipulation of lamin levels by the addition of recombinant proteins. We find that nuclear growth and size are sensitive to the levels of nuclear lamins, with low and high concentrations increasing and decreasing nuclear size, respectively. Interestingly, each type of lamin that we tested (lamins B1, B2, B3, and A) similarly affected nuclear size whether added alone or in combination, suggesting that total lamin concentration, and not lamin type, is more critical to determining nuclear size. Furthermore, we show that altering lamin levels in vivo, both in Xenopus embryos and mammalian tissue culture cells, also impacts nuclear size. These results have implications for normal development and carcinogenesis where both nuclear size and lamin expression levels change. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Advanced Manufacturing Technologies (AMT): Additive Manufactured Hot Fire Planning and Testing in GRC Cell 32 Project

    NASA Technical Reports Server (NTRS)

    Fikes, John C.

    2014-01-01

    The objective of this project is to hot fire test an additively manufactured thrust chamber assembly TCA (injector and thrust chamber). GRC will install the additively manufactured Inconel 625 injector, two additively manufactured (SLM) water cooled Cu-Cr thrust chamber barrels and one additively manufactured (SLM) water cooled Cu-Cr thrust chamber nozzle on the test stand in Cell 32 and perform hot fire testing of the integrated TCA.

  1. Equation of state of hot polarized nuclear matter using the generalized Skyrme interaction

    NASA Astrophysics Data System (ADS)

    Abd-Alla, M.; Hager, S. A.

    2000-04-01

    We used the generalized Skyrme potential to study the equation of state of polarized nuclear matter in the frame of the Thomas-Fermi model. The critical temperature of the liquid-gas phase transition is found to be Tc=16.2 MeV. This critical temperature was found to decease with the asymmetry, spin, and spin-isospin excess parameters. The isothermal compressibility of polarized nuclear matter was also studied. The volume compressibility Kv was found to decrease with temperature. The symmetry compressibility Kx, the spin symmetry compressibility Ky, and the spin-isospin symmetry compressibility Kz were found to have a little increasing behavior with temperature.

  2. Nuclear microscopy of sperm cell elemental structure

    SciTech Connect

    Bench, G.S.

    1994-12-31

    Theories have suggested that there is a link between protamine concentrations in individual sperm and sperm fertility. At present, biochemical analyses have only been performed on bulk populations and existing methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. As part of an investigation into male sperm fertility, nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the ratio of Phosphorus to Sulfur the authors have been able to determine the amount of protamine 1 and protamine 2 in individual cells from bulk fertile samples of bull and mouse sperm. Preliminary results show that, for each species, the relative amounts of protamine 1 and protamine 2 in morphologically normal sperm agree well with expected values.

  3. CALIBRATION AND HOT TESTING OF THE ADVANCED NUCLEAR MEASUREMENT SYSTEMS USED FOR WASTE CHARACTERIZATION IN COGEMA'S NEW ACC COMPACTION FACILITY

    SciTech Connect

    Toubon, H.; Vuillier; Gain, T.; Huver, M.

    2003-02-27

    Spent nuclear fuel from commercial power reactors is reprocessed at the COGEMA plant in La Hague. After shearing and dissolution of the fuel assemblies, the hulls and nozzles are sent to COGEMA's new compaction facility (ACC) to reduce the final volume of waste. Technological waste generated in the reprocessing plant is also sent to the ACC facility. Compacted waste is characterized by two measurement stations: a gamma spectrometry station and an active and passive neutron measurement station. The main purpose of these measurement stations is to determine the guaranteed nuclear parameters of the compacted waste and their associated uncertainties: (1) total U and Pu masses, (2) Pu, Cm, and total alpha activities, (3) 137Cs, 90Sr-90Y,241Pu beta activities, (4) decay heat. After giving a description of the measurement stations, this paper will describe the qualification tests performed in the context of the ACC project. The extensive calibration tests performed on site with different sources and different waste matrices will be described (approximately 500 neutron and gamma experiments). Hot tests that were conducted after hot start-up at the end of 2001 and prior to the start of commercial operation will be also presented. A number of drums produced by the upstream facilities were introduced one by one into the ACC facility in order to avoid mixing of different fuel assemblies. This procedure allows comparison between characterization performed in the upstream facilities on the basis of fuel data available before processing and the measurements performed on the new ACC stations. These comparisons showed good agreement between the different methods of characterization and thus validated the innovative technologies and methods used by COGEMA for compacted waste generated by the ACC facility.

  4. Hot Hydrogen Testing of Tungsten-Uranium Dioxide (W-UO2) CERMET Fuel Materials for Nuclear Thermal Propulsion

    NASA Technical Reports Server (NTRS)

    Hickman, Robert; Broadway, Jeramie

    2014-01-01

    CERMET fuel materials are being developed at the NASA Marshall Space Flight Center for a Nuclear Cryogenic Propulsion Stage. Recent work has resulted in the development and demonstration of a Compact Fuel Element Environmental Test (CFEET) System that is capable of subjecting depleted uranium fuel material samples to hot hydrogen. A critical obstacle to the development of an NCPS engine is the high-cost and safety concerns associated with developmental testing in nuclear environments. The purpose of this testing capability is to enable low-cost screening of candidate materials, fabrication processes, and further validation of concepts. The CERMET samples consist of depleted uranium dioxide (UO2) fuel particles in a tungsten metal matrix, which has been demonstrated on previous programs to provide improved performance and retention of fission products1. Numerous past programs have utilized hot hydrogen furnace testing to develop and evaluate fuel materials. The testing provides a reasonable simulation of temperature and thermal stress effects in a flowing hydrogen environment. Though no information is gained about radiation damage, the furnace testing is extremely valuable for development and verification of fuel element materials and processes. The current work includes testing of subscale W-UO2 slugs to evaluate fuel loss and stability. The materials are then fabricated into samples with seven cooling channels to test a more representative section of a fuel element. Several iterations of testing are being performed to evaluate fuel mass loss impacts from density, microstructure, fuel particle size and shape, chemistry, claddings, particle coatings, and stabilizers. The fuel materials and forms being evaluated on this effort have all been demonstrated to control fuel migration and loss. The objective is to verify performance improvements of the various materials and process options prior to expensive full scale fabrication and testing. Post test analysis will

  5. HOT CELL BUILDING, TRA632, INTERIOR. WRIGHT 3TON HOIST ON EAST ...

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

    HOT CELL BUILDING, TRA-632, INTERIOR. WRIGHT 3-TON HOIST ON EAST SIDE OF CELL 2. SIGN AT LEFT OF VIEW SAYS, "...DO NOT BRING FISSILE MATERIAL INTO AREA WITHOUT APPROVAL." CAMERA FACES NORTHWEST. INL NEGATIVE NO. HD46-29-2. Mike Crane, Photographer, 2/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  6. Hot-carrier solar cell spectral insensitivity: Why develop the hot-carrier solar cell when we have multi-junction devices?

    NASA Astrophysics Data System (ADS)

    Hirst, Louise C.; Lumb, Matthew P.; Hoheisel, Raymond; Philipps, Simon P.; Bett, Andreas W.; Walters, Robert J.

    2014-03-01

    The hot carrier solar cell (HCSC) offers one route to high efficiency solar energy conversion and has similar fundamental limiting efficiency to multi-junction (MJ) solar cells however, the HCSC is at a much earlier stage of development. We discuss the unique features of the HCSC which distinguish it from other PV technologies, providing motivation for development. We consider the potential for a low concentration hot-carrier enhanced single-junction solar cell, enabled by field enhancing cell architectures. To support this we experimentally show that changing sample geometry to increase carrier density, while keeping phononic and electronic properties constant, substantially reduces hot-carrier themalization coefficient. Such a scheme might have similar applications to todays high efficiency single-junction devices while allowing from some intrinsic efficiency enhancement. We also use spectral data simulated using SMARTS to identify HCSC spectral insensitivity relative to MJ devices. Spectral insensitivity increases annual energy yield relative to laboratory test efficiency, reducing the cost of PV power generation. There are also several practical advantages: a single device design will operate optimally in a variety of locations and solar power stations are less reliant of accurate, long-range atmospheric simulation to achieve energy yield targets.

  7. HOT CELL BUILDING, TRA632. FLOOR PLAN OF EXPANSION SHOWS LOCATION ...

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

    HOT CELL BUILDING, TRA-632. FLOOR PLAN OF EXPANSION SHOWS LOCATION OF NEW CELLS, "HEAVY" CELL AT WEST END, "LIGHT" CELLS AT EAST. MOCK-UP AND STORAGE AREAS IN SOUTH HALF OF FLOOR. H.K. FERGUSON 895-MTR-ETR-632-A1, 12/1958. INL INDEX NO. 531-0632-00-279-101924, REV. 4. - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  8. Reversible electron-hole separation in a hot carrier solar cell

    NASA Astrophysics Data System (ADS)

    Linke, Heiner

    Hot-carrier solar cells are envisioned to utilize energy filtering to extract power from photogenerated electron-hole pairs before they thermalize with the lattice, and thus potentially offer higher power conversion efficiency compared to conventional, single absorber solar cells. The efficiency of hot-carrier solar cells can be expected to strongly depend on the details of the energy filtering process, a relationship which to date has not been satisfactorily explored. Here, we establish the conditions under which electron-hole separation in hot-carrier solar cells can occur reversibly, that is, at maximum energy conversion efficiency. We find that, under specific conditions, the energy conversion efficiency of a hot-carrier solar cell can exceed the Carnot limit set by the intra-device temperature gradient alone, due to the additional contribution of the quasi-Fermi level splitting in the absorber. To achieve this, we consider a highly selective energy filter such as a quantum dot embedded into a one-dimensional conductor. We also establish that the open-circuit voltage of a hot-carrier solar cell is not limited by the band gap of the absorber, due to the additional thermoelectric contribution to the voltage. Additionally, we find that a hot-carrier solar cell can be operated in reverse as a thermally driven solid-state light emitter. In addition this theoretical analysis, I will also report on first experimental results in a nanowire-based energy filter device. Ref: S Limpert, S Bremner, and H Linke, New J. Phys 17, 095004 (2015)

  9. Self-consistent calculation of the nuclear composition in hot and dense stellar matter

    NASA Astrophysics Data System (ADS)

    Furusawa, Shun; Mishustin, Igor

    2017-03-01

    We investigate the mass fractions and in-medium properties of heavy nuclei in stellar matter at characteristic densities and temperatures for supernova (SN) explosions. The individual nuclei are described within the compressible liquid-drop model taking into account modifications of bulk, surface, and Coulomb energies. The equilibrium properties of nuclei and the full ensemble of heavy nuclei are calculated self-consistently. It is found that heavy nuclei in the ensemble are either compressed or decompressed depending on the isospin asymmetry of the system. The compression or decompression has a little influence on the binding energies, total mass fractions, and average mass numbers of heavy nuclei, although the equilibrium densities of individual nuclei themselves are changed appreciably above one-hundredth of normal nuclear density. We find that nuclear structure in the single-nucleus approximation deviates from the actual one obtained in the multinucleus description, since the density of free nucleons is different between these two descriptions. This study indicates that a multinucleus description is required to realistically account for in-medium effects on the nuclear structure in supernova matter.

  10. Imaging of spatially extended hot spots with coded apertures for intra-operative nuclear medicine applications

    NASA Astrophysics Data System (ADS)

    Kaissas, I.; Papadimitropoulos, C.; Potiriadis, C.; Karafasoulis, K.; Loukas, D.; Lambropoulos, C. P.

    2017-01-01

    Coded aperture imaging transcends planar imaging with conventional collimators in efficiency and Field of View (FOV). We present experimental results for the detection of 141 keV and 122 keV γ-photons emitted by uniformly extended 99mTc and 57Co hot-spots along with simulations of uniformly and normally extended 99mTc hot-spots. These results prove that the method can be used for intra-operative imaging of radio-traced sentinel nodes and thyroid remnants. The study is performed using a setup of two gamma cameras, each consisting of a coded-aperture (or mask) of Modified Uniformly Redundant Array (MURA) of rank 19 positioned on top of a CdTe detector. The detector pixel pitch is 350 μm and its active area is 4.4 × 4.4 cm2, while the mask element size is 1.7 mm. The detectable photon energy ranges from 15 keV up to 200 keV with an energy resolution of 3-4 keV FWHM. Triangulation is exploited to estimate the 3D spatial coordinates of the radioactive spots within the system FOV. Two extended sources, with uniform distributed activity (11 and 24 mm in diameter, respectively), positioned at 16 cm from the system and with 3 cm distance between their centers, can be resolved and localized with accuracy better than 5%. The results indicate that the estimated positions of spatially extended sources lay within their volume size and that neighboring sources, even with a low level of radioactivity, such as 30 MBq, can be clearly distinguished with an acquisition time about 3 seconds.

  11. Nuclear Star Formation in the Hot-Spot Galaxy NGC 2903

    NASA Technical Reports Server (NTRS)

    Alonso-Herrero, A.; Ryder, S. D.; Knapen, J. H.

    1994-01-01

    We present high-resolution near-infrared imaging obtained using adaptive optics and HST/NICMOS and ground-based spectroscopy of the hot-spot galaxy NGC 2903. Our near-infrared resolution imaging enables us to resolve the infrared hot spots into individual young stellar clusters or groups of these. The spatial distribution of the stellar clusters is not coincident with that of the bright H II regions, as revealed by the HST/NICMOS Pace image. Overall, the circumnuclear star formation in NGC 2903 shows a ring-like morphology with an approximate diameter of 625 pc. The SF properties of the stellar clusters and H II regions have been studied using the photometric and spectroscopic information in conjunction with evolutionary synthesis models. The population of bright stellar clusters shows a very narrow range of ages, 4 to 7 x 10(exp 6) yr after the peak of star formation, or absolute ages 6.5 to 9.5 x 10(exp 6) yr (for the assumed short-duration Gaussian bursts), and luminosities similar to the clusters found in the Antennae interacting galaxy. This population of young stellar clusters accounts for some 7 - 12% of the total stellar mass in the central 625 pc of NGC 2903. The H II regions in the ring of star formation have luminosities close to that of the super-giant H II region 30 Doradus, they are younger than the stellar clusters, and will probably evolve into bright infrared stellar clusters similar to those observed today. We find that the star formation efficiency in the central regions of NGC 2903 is higher than in normal galaxies, approaching the lower end of infrared luminous galaxies.

  12. Tandem-structured, hot electron based photovoltaic cell with double Schottky barriers.

    PubMed

    Lee, Young Keun; Lee, Hyosun; Park, Jeong Young

    2014-04-03

    We demonstrate a tandem-structured, hot electron based photovoltaic cell with double Schottky barriers. The tandem-structured, hot electron based photovoltaic cell is composed of two metal/semiconductor interfaces. Two types of tandem cells were fabricated using TiO2/Au/Si and TiO2/Au/TiO2, and photocurrent enhancement was detected. The double Schottky barriers lead to an additional pathway for harvesting hot electrons, which is enhanced through multiple reflections between the two barriers with different energy ranges. In addition, light absorption is improved by the band-to-band excitation of both semiconductors with different band gaps. Short-circuit current and energy conversion efficiency of the tandem-structured TiO2/Au/Si increased by 86% and 70%, respectively, compared with Au/Si metal/semiconductor nanodiodes, showing an overall solar energy conversion efficiency of 5.3%.

  13. Tandem-structured, hot electron based photovoltaic cell with double Schottky barriers

    PubMed Central

    Lee, Young Keun; Lee, Hyosun; Park, Jeong Young

    2014-01-01

    We demonstrate a tandem-structured, hot electron based photovoltaic cell with double Schottky barriers. The tandem-structured, hot electron based photovoltaic cell is composed of two metal/semiconductor interfaces. Two types of tandem cells were fabricated using TiO2/Au/Si and TiO2/Au/TiO2, and photocurrent enhancement was detected. The double Schottky barriers lead to an additional pathway for harvesting hot electrons, which is enhanced through multiple reflections between the two barriers with different energy ranges. In addition, light absorption is improved by the band-to-band excitation of both semiconductors with different band gaps. Short-circuit current and energy conversion efficiency of the tandem-structured TiO2/Au/Si increased by 86% and 70%, respectively, compared with Au/Si metal/semiconductor nanodiodes, showing an overall solar energy conversion efficiency of 5.3%. PMID:24694838

  14. Removal of an acid fume system contaminated with perchlorates located within hot cell

    SciTech Connect

    Rosenberg, K.E.; Henslee, S.P.; Vroman, W.R.; Krsul, J.R.; Michelbacher, J.A.; Knighton, G.C.

    1992-09-01

    An add scrubbing system located within the confines of a highly radioactive hot cell at Argonne National Laboratory-West (ANL-W) was remotely removed. The acid scrubbing system was routinely used for the dissolution of irradiated reactor fuel samples and structural materials. Perchloric acid was one of the acids used in the dissolution process and remained in the system with its inherent risks. Personnel could not enter the hot cell to perform the dismantling of the acid scabbing system due to the high radiation field and the explosion potential associated with the perchlorates. A robot was designed and built at ANL-W and used to dismantle the system without the need for personnel entry into the hot cell. The robot was also used for size reduction of removed components and loading of the removed components into waste containers.

  15. Methods for Assessing Nuclear Rotation and Nuclear Positioning in Developing Skeletal Muscle Cells

    PubMed Central

    Wilson, Meredith H.; Bray, Matthew G.; Holzbaur, Erika L.F.

    2016-01-01

    Summary Skeletal muscle cells are large syncytia, containing hundreds of nuclei positioned regularly along the length of the fiber. During development, nuclei are actively distributed throughout the myotube by the microtubule motor proteins, kinesin-1 and cytoplasmic dynein. Nuclear movement consists of translocation along the long axis of the cell concurrent with three-dimensional rotation of nuclei. In this chapter we describe methods for quantitatively assessing the speed of nuclear rotation in cultured myotubes using live-cell imaging techniques coupled with rigid-body kinematic analyses. Additionally we provide protocols for analyzing nuclear distribution in myotubes. PMID:27147049

  16. Remote maintenance for a new generation of hot cells

    SciTech Connect

    Feldman, M.J.; Grant, N.R.

    1987-01-01

    For several years the Consolidated Fuel Reprocessing Program (CFRP) at Oak Ridge National Laboratory (ORNL) has been developing facility concepts, designing specialized equipment, and testing prototypical hardware for reprocessing spent fuel from fast breeder reactors. The major facility conceptual design, the Hot Experimental Facility, was based on total remote maintenance to increase plant availability and to reduce radiation exposure. This thrust included designing modular equipment to facilitate maintenance and the manipulation necessary to accomplish maintenance. Included in the design repetoire was the development effort in advanced servomanipulator systems, a remote sampling system, television viewing, and a transporter for manipulator positioning. Demonstration of these developed items is currently ongoing, and the technology is available for applications where production operations in highly radioactive environments are required.

  17. Nuclear protein import is reduced in cells expressing nuclear envelopathy-causing lamin A mutants

    SciTech Connect

    Busch, Albert; Kiel, Tilman; Heupel, Wolfgang-M.; Wehnert, Manfred; Huebner, Stefan

    2009-08-15

    Lamins, which form the nuclear lamina, not only constitute an important determinant of nuclear architecture, but additionally play essential roles in many nuclear functions. Mutations in A-type lamins cause a wide range of human genetic disorders (laminopathies). The importance of lamin A (LaA) in the spatial arrangement of nuclear pore complexes (NPCs) prompted us to study the role of LaA mutants in nuclear protein transport. Two mutants, causing prenatal skin disease restrictive dermopathy (RD) and the premature aging disease Hutchinson Gilford progeria syndrome, were used for expression in HeLa cells to investigate their impact on the subcellular localization of NPC-associated proteins and nuclear protein import. Furthermore, dynamics of the LaA mutants within the nuclear lamina were studied. We observed affected localization of NPC-associated proteins, diminished lamina dynamics for both LaA mutants and reduced nuclear import of representative cargo molecules. Intriguingly, both LaA mutants displayed similar effects on nuclear morphology and functions, despite their differences in disease severity. Reduced nuclear protein import was also seen in RD fibroblasts and impaired lamina dynamics for the nucleoporin Nup153. Our data thus represent the first study of a direct link between LaA mutant expression and reduced nuclear protein import.

  18. HOT CELL BUILDING, TRA632. CONTEXTUAL VIEW ALONG WALLEYE AVENUE, CAMERA ...

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

    HOT CELL BUILDING, TRA-632. CONTEXTUAL VIEW ALONG WALLEYE AVENUE, CAMERA FACING EASTERLY. HOT CELL BUILDING IS AT CENTER LEFT OF VIEW; THE LOW-BAY PROJECTION WITH LADDER IS THE TEST TRAIN ASSEMBLY FACILITY, ADDED IN 1968. MTR BUILDING IS IN LEFT OF VIEW. HIGH-BAY BUILDING AT RIGHT IS THE ENGINEERING TEST REACTOR BUILDING, TRA-642. INL NEGATIVE NO. HD46-32-1. Mike Crane, Photographer, 4/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  19. Spin-polarized lithium diffusion in a glass hot-vapor cell

    NASA Astrophysics Data System (ADS)

    Ishikawa, Kiyoshi

    2016-08-01

    We report diffusion coefficients of optically pumped lithium atoms in helium buffer gas. The free-induction decay and the spin-echo signals of ground-state atoms were optically detected in an external magnetic field with the addition of field gradient. Lithium hot vapor was produced in a borosilicate-glass cell at a temperature between 290 and 360°C. The simple setup using the glass cells enabled lithium atomic spectroscopy in a similar way to other alkali-metal atoms and study of the collisional properties of lithium atoms in a hot-vapor phase.

  20. Nonextensive statistical effects and strangeness production in hot and dense nuclear matter

    NASA Astrophysics Data System (ADS)

    Lavagno, A.; Pigato, D.

    2012-12-01

    By means of an effective relativistic nuclear equation of state in the framework of the nonextensive statistical mechanics, characterized by power-law quantum distributions, we study the phase transition from hadronic matter to quark-gluon plasma at finite temperature and baryon density. The analysis is performed by requiring the Gibbs conditions on the global conservation of baryon number, electric charge fraction and zero net strangeness. We show that nonextensive statistical effects strongly influence the strangeness production during the pure hadronic phase and the hadron-quark-gluon mixed phase transition, also for small deviations from the standard Boltzmann-Gibbs statistics.

  1. Nuclear reprogramming and its role in vascular smooth muscle cells.

    PubMed

    Zaina, Silvio; del Pilar Valencia-Morales, Maria; Tristán-Flores, Fabiola E; Lund, Gertrud

    2013-09-01

    In general terms, "nuclear reprogramming" refers to a change in gene expression profile that results in a significant switch in cellular phenotype. Nuclear reprogramming was first addressed by pioneering studies of cell differentiation during embryonic development. In recent years, nuclear reprogramming has been studied in great detail in the context of experimentally controlled dedifferentiation and transdifferentiation of mammalian cells for therapeutic purposes. In this review, we present a perspective on nuclear reprogramming in the context of spontaneous, pathophysiological phenotypic switch of vascular cells occurring in the atherosclerotic lesion. In particular, we focus on the current knowledge of epigenetic mechanisms participating in the extraordinary flexibility of the gene expression profile of vascular smooth muscle cells and other cell types participating in atherogenesis. Understanding how epigenetic changes participate in vascular cell plasticity may lead to effective therapies based on the remodelling of the vascular architecture.

  2. Nuclear PTEN levels and G2 progression in melanoma cells

    PubMed Central

    Jacob, Abraham I.; Romigh, Todd; Waite, Kristin A.; Eng, Charis

    2009-01-01

    The phosphatase and tensin homolog (PTEN) exerts its function, in part, by negatively regulating the well-known phosphatidylinositol-3-kinase/AKT signaling pathway. Previous histological work has suggested that alterations in the nuclear/cytoplasmic compartmentalization of PTEN may play a role in the development and progression of melanoma. In this study, we examined the nuclear/cytoplasmic compartmentalization of PTEN in melanoma cell lines and its correlation with the cell cycle. Studies were performed in melanoma cells lines using classic cell biological techniques. In contrast to breast cancer cell lines, we found that increased levels of nuclear PTEN levels correlate with G2 rather than with G1 arrest. In WM164 and SKmel28 cells, overexpression of PTEN protein did not significantly increase the number of cells in the G2 phase. Differential CDC2 phosphorylation levels in cells that overexpressed PTEN compared with those where PTEN was downregulated suggest some involvement of PTEN in G2 checkpoint regulation. The data suggest that although nuclear PTEN levels correlate with the G2 phase, the role of PTEN in modulating G2/M arrest is not limiting. Further, the specific cell cycle phase regulated by nuclear PTEN is cell-type dependent. Taken together, our observations suggest that in melanoma, nuclear PTEN is involved in G2 progression possibly through the modulation of CDC2, opening up a new arena for investigation. PMID:19478684

  3. Third generation hot carrier solar cells: paths towards innovative energy contacts structures

    NASA Astrophysics Data System (ADS)

    Gibelli, Francois; Julian, Anatole; Jehl Li Kao, Zacharie; Guillemoles, Jean-François

    2016-03-01

    The hot carrier solar cell is a very promising clean energy technology, with the potential to achieve high conversion yields with constrained costs. Due to the hot carrier effect, the estimation of the achievable voltage needs some theoretical developments. The classical approach is to consider isentropic energy selective contacts, converting the excess of kinetic energy of the hot carriers into electrical potential energy. Here we show the differences between the ideal case of isentropic contacts and the more realistic one, with an output voltage of the cell depending on the transmission function. We particularly emphasize the importance of the transmission function of the contact on both output current and output voltage, modifying thereby the classical view of the output power dependence on the transmission function.

  4. Intermediate filaments promote nuclear mechanical constraints during somatic cell nuclear transfer in the mouse.

    PubMed

    Gall, Laurence; Brochard, Vincent; Ruffini, Sylvie; Laffont, Ludivine; Fleurot, Renaud; Lavin, Tiphaine Aguirre; Jouneau, Alice; Beaujean, Nathalie

    2012-12-01

    The somatic cell nuclear transfer (SCNT) procedure requires nuclear remodeling to return differentiated somatic nuclei to the totipotent undifferentiated stage. We hypothesize that mechanical constraints might occur upon SCNT and thereby affect nuclear remodeling. Therefore, we analyzed the nuclear structures upon SCNT using as donors either wild-type fibroblasts with a dense vimentin network or vimentin-deprived cells [embryonic stem cells (ESCs) and fibroblasts invalidated for vimetin]. We demonstrated that following nuclear transfer of wild-type fibroblasts, vimentin intermediate filaments (IFs) persisted around the transplanted nuclei and 88% of them presented severe distortions. We also showed that the presence of vimentin filaments in the reconstructed embryos was correlated with DNA damage, as evidenced by γH2A.X foci. On the other hand, when ESCs or vimentin-null (Vim(-/-)) fibroblasts devoid of IFs were used as nuclear donors, no nuclear distortion and less DNA damage were observed. Altogether we believe that the introduction of vimentin into recipient oocytes during SCNT induces a mechanical constraint on the transplanted nucleus that is responsible for nuclear distortions and DNA damage. This could lead to incomplete reprogramming that would be detrimental to further embryonic development.

  5. Emerging role for nuclear rotation and orientation in cell migration

    PubMed Central

    Maninová, Miloslava; Iwanicki, Marcin P; Vomastek, Tomáš

    2014-01-01

    Nucleus movement, positioning, and orientation is precisely specified and actively regulated within cells, and it plays a critical role in many cellular and developmental processes. Mutation of proteins that regulate the nucleus anchoring and movement lead to diverse pathologies, laminopathies in particular, suggesting that the nucleus correct positioning and movement is essential for proper cellular function. In motile cells that polarize toward the direction of migration, the nucleus undergoes controlled rotation promoting the alignment of the nucleus with the axis of migration. Such spatial organization of the cell appears to be optimal for the cell migration. Nuclear reorientation requires the cytoskeleton to be anchored to the nuclear envelope, which exerts pulling or pushing torque on the nucleus. Here we discuss the possible molecular mechanisms regulating the nuclear rotation and reorientation and the significance of this type of nuclear movement for cell migration. PMID:24589621

  6. Hot topics in stem cells and self-renewal: 2010.

    PubMed

    Sharpless, Norman E

    2010-08-01

    In many tissues, mammalian aging is associated with a decline in the replicative and functional capacity of somatic stem cells and other self-renewing compartments. Understanding the basis of this decline is a major goal of aging research. In particular, therapeutic approaches to ameliorate or reverse the age-associated loss of stem function could be of use in clinical geriatrics. Such approaches include attempts to protect stem cells from age-promoting damage, to 'rejuvenate' stem cells through the use of pharmacologic agents that mitigate aging-induced alterations in signaling, and to replace lost stem cells through regenerative medicine approaches. Some headway has been made in each of these arenas over the last 18 months including advances in the production of donor-specific totipotent stem cells through induced pluripotency (iPS), gains in our understanding of how tumor suppressor signaling is controlled in self-renewing compartments to regulate aging, and further demonstration of extracellular 'milieu' factors that perturb stem cell function with age. This period has also been marked by the recent award of the Nobel Prize in Physiology or Medicine for elucidation of telomeres and telomerase, a topic of critical importance to stem cell aging.

  7. Nuclear transfer to study the nuclear reprogramming of human stem cells.

    PubMed

    Saito, Shigeo; Sawai, Ken; Murayama, Yoshinobu; Fukuda, Keiichi; Yokoyama, Kazunari

    2008-01-01

    Research of stem cells will enable us to understand the development and function of tissues and organs in mammals. The ability to induce regeneration of new tissues from embryonic stem (ES) cells derived from cloned blastocysts via nuclear transfer can be expected in the not-too-distant future. The fact that there is no way except nuclear cloning for the return of differentiated cells to undifferentiated cells remains an interesting problem to be solved. We describe protocols for the production of cloned calves from bovine ES cells to study nuclear reprogramming ability of stem cells. The frequency of term pregnancies for blastocysts from ES cells is higher than those of early pregnancies and maintained pregnancies after nuclear transfer with bovine somatic cells. We also describe protocols for gene introduction into bovine ES cells in vitro, particularly the human leukocyte antigens (HLA). Bovine ES cells provide a powerful tool for the generation of transgenic clonal offspring. This technique, when perfected for humans, may be critical for neural stem cell transplantation.

  8. HOT CELL BUILDING, TRA632. SOUTH FACADE. CAMERA FACING NORTHEAST AND ...

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

    HOT CELL BUILDING, TRA-632. SOUTH FACADE. CAMERA FACING NORTHEAST AND DETAIL OF TEST TRAIN ASSEMBLY FACILITY ON WEST END. SIGN SAYS "ISM, DOING WORK SAFELY." INL NEGATIVE NO. HD46-32-2. Mike Crane, Photographer, 4/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  9. A&M. TAN607. Special service cubicle (hot cell). Details include Zpipe ...

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

    A&M. TAN-607. Special service cubicle (hot cell). Details include Z-pipe and stepped plug penetrations through shielding wall. Ralph M. Parsons 902-3-ANP-607-A116. Date: December 1952. Approved by INEEL Classification Office for public release. INEEL index code no. 034-0607-693-106767 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  10. A&M. Hot cell annex (TAN633). Camera facing east. Construction view ...

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

    A&M. Hot cell annex (TAN-633). Camera facing east. Construction view of north and west walls. Photographer: Jack L. Anderson. Date: October 23, 1957. INEEL negative no. 57-5336 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  11. A&M. Hot cell annex (TAN633) contextual view also showing east ...

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

    A&M. Hot cell annex (TAN-633) contextual view also showing east facade. Camera facing west. Note corridor connecting annex to pool area of TAN-607. Pumice block walls. Date: March 2004. INEEL negative no. HD-39-2-2 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  12. HOT CELL BUILDING, TRA632. ELEVATIONS. PUMICE BLOCK WALLS. BLOWER AND ...

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

    HOT CELL BUILDING, TRA-632. ELEVATIONS. PUMICE BLOCK WALLS. BLOWER AND FILTER LOFT PLATFORM AND LADDER ON EAST SIDE. IDAHO OPERATIONS OFFICE MTR-632-IDO-4, 11/1952. INL INDEX NO. 531-0632-00-396-110563, REV. 2. - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  13. VIEW OF FECF HOT CELL OF FUEL STORAGE BUILDING (CPP603). ...

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

    VIEW OF FECF HOT CELL OF FUEL STORAGE BUILDING (CPP-603). PHOTO TAKEN LOOKING NORHTWEST. INL PHOTO NUMBER HD-54-18-3. Mike Crane, Photographer, 8/2005 - Idaho National Engineering Laboratory, Idaho Chemical Processing Plant, Fuel Reprocessing Complex, Scoville, Butte County, ID

  14. 114. ARAI Hot cell (ARA626) Building details of fuel storage ...

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

    114. ARA-I Hot cell (ARA-626) Building details of fuel storage pit in plan and section. Spaces shown for 20 elements. Norman Engineering Company: 961-area/SF-626-S-4. Date: January 1959. Ineel index code no. 068-0626-60-613-102752. - Idaho National Engineering Laboratory, Army Reactors Experimental Area, Scoville, Butte County, ID

  15. 111. ARAI Hot cell (ARA626) Building elevations of north, south, ...

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

    111. ARA-I Hot cell (ARA-626) Building elevations of north, south, east, and west sides. Includes details of personnel decontamination area, dark room, and other features. Norman Engineering Company: 961-area/SF-626-A-3. Date: January 1959. Ineel index code no. 068-0626-00-613-102723. - Idaho National Engineering Laboratory, Army Reactors Experimental Area, Scoville, Butte County, ID

  16. 112. ARAI Hot cell (ARA626) Building roof plan and details ...

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

    112. ARA-I Hot cell (ARA-626) Building roof plan and details of roof ventilating equipment and parapet. Norman Engineering Company: 961-area/SF-626-A-2. Date: January 1959. Ineel index code no. 068-0626-00-613-102722. - Idaho National Engineering Laboratory, Army Reactors Experimental Area, Scoville, Butte County, ID

  17. 115. ARAI Details of hot cell section of building ARA626. ...

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

    115. ARA-I Details of hot cell section of building ARA-626. Shows location of high density concrete, viewing windows, filters, monorail crane, bridge crane, and other details. Norman Engineering Company 961-area/SF-626-MS-1. Date: January 1959. Ineel index code no. 068-0626-40-613-102737. - Idaho National Engineering Laboratory, Army Reactors Experimental Area, Scoville, Butte County, ID

  18. HOT CELL BUILDING, TRA632. SHIELDED WINDOWS HAVE BEEN INSTALLED. MANIPULATORS ...

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

    HOT CELL BUILDING, TRA-632. SHIELDED WINDOWS HAVE BEEN INSTALLED. MANIPULATORS AWAIT ATTACHMENT TO HAND CONTROLS. INL NEGATIVE NO. 9001. Unknown Photographer, photo is identified as taken 10/28/1953, but it may be an error as it shows progress since ID-33-G-266 of same date. - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  19. HOT CELL BUILDING, TRA632. INSTRUMENT FITTINGS, MASTER/SLAVE MANIPULATOR, "POT LID ...

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

    HOT CELL BUILDING, TRA-632. INSTRUMENT FITTINGS, MASTER/SLAVE MANIPULATOR, "POT LID CRANE." IDAHO OPERATIONS OFFICE MTR-632-IDO-16, 11/1952. INL INDEX NO. 531-0632-40-396-110574, REV. 2. - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  20. Extreme nuclear shapes examined via giant dipole resonance lineshapes in hot light-mass systems

    SciTech Connect

    Pandit, Deepak; Mukhopadhyay, S.; Pal, Surajit; Bhattacharya, S.; Bhattacharya, C.; Banerjee, K.; Kundu, S.; Rana, T. K.; Dey, A.; Mukherjee, G.; Ghosh, T.; Banerjee, S. R.; De, A.; Gupta, D.

    2010-06-15

    The influence of alpha clustering on nuclear reaction dynamics is investigated using the giant dipole resonance (GDR) lineshape studies in the reactions {sup 20}Ne (E{sub lab}=145,160 MeV) + {sup 12}C and {sup 20}Ne (E{sub lab}=160 MeV) + {sup 27}Al, populating {sup 32}S and {sup 47}V, respectively. The GDR lineshapes from the two systems are remarkably different from each other. Whereas, the non-alpha-like {sup 47}V undergoes Jacobi shape transition and matches exceptionally well with the theoretical GDR lineshape estimated under the framework rotating liquid drop model (RLDM) and thermal shape fluctuation model (TSFM) signifying shape equilibration, for the alpha cluster {sup 32}S an extended prolate kind of shape is observed. This unusual deformation, seen directly via gamma decay for the first time, is predicted to be due to the formation of orbiting dinuclear configuration or molecular structure of {sup 16}O + {sup 16}O in the {sup 32}S superdeformed band.

  1. Indications for a critical end point in the phase diagram for hot and dense nuclear matter.

    PubMed

    Lacey, Roy A

    2015-04-10

    Excitation functions for the Gaussian emission source radii difference (R_{out}^{2}-R_{side}^{2}) obtained from two-pion interferometry measurements in Au+Au (sqrt[s_{NN}]=7.7-200  GeV) and Pb+Pb (sqrt[s_{NN}]=2.76  TeV) collisions are studied for a broad range of collision centralities. The observed nonmonotonic excitation functions validate the finite-size scaling patterns expected for the deconfinement phase transition and the critical end point (CEP), in the temperature versus baryon chemical potential (T,μ_{B}) plane of the nuclear matter phase diagram. A finite-size scaling (FSS) analysis of these data suggests a second order phase transition with the estimates T^{cep}∼165  MeV and μ_{B}^{cep}∼95  MeV for the location of the critical end point. The critical exponents (ν≈0.66 and γ≈1.2) extracted via the same FSS analysis place this CEP in the 3D Ising model universality class.

  2. Nuclear envelope rupture and repair during cancer cell migration.

    PubMed

    Denais, Celine M; Gilbert, Rachel M; Isermann, Philipp; McGregor, Alexandra L; te Lindert, Mariska; Weigelin, Bettina; Davidson, Patricia M; Friedl, Peter; Wolf, Katarina; Lammerding, Jan

    2016-04-15

    During cancer metastasis, tumor cells penetrate tissues through tight interstitial spaces, which requires extensive deformation of the cell and its nucleus. Here, we investigated mammalian tumor cell migration in confining microenvironments in vitro and in vivo. Nuclear deformation caused localized loss of nuclear envelope (NE) integrity, which led to the uncontrolled exchange of nucleo-cytoplasmic content, herniation of chromatin across the NE, and DNA damage. The incidence of NE rupture increased with cell confinement and with depletion of nuclear lamins, NE proteins that structurally support the nucleus. Cells restored NE integrity using components of the endosomal sorting complexes required for transport III (ESCRT III) machinery. Our findings indicate that cell migration incurs substantial physical stress on the NE and its content and requires efficient NE and DNA damage repair for cell survival. Copyright © 2016, American Association for the Advancement of Science.

  3. Importin-7 Mediates Nuclear Trafficking of DNA in Mammalian Cells

    PubMed Central

    Dhanoya, Arjun; Wang, Tse; Keshavarz-Moore, Eli; Fassati, Ariberto; Chain, Benjamin M

    2013-01-01

    Eukaryotic cells have the ability to uptake and transport endogenous and exogenous DNA in their nuclei, however little is known about the specific pathways involved. Here we show that the nuclear transport receptor importin 7 (imp7) supports nuclear import of supercoiled plasmid DNA and human mitochondrial DNA in a Ran and energy-dependent way. The imp7-dependent pathway was specifically competed by excess DNA but not by excess of maltose-binding protein fused with the classical nuclear localizing signal (NLS) or the M9 peptides. Transport of DNA molecules complexed with poly-l-lysine was impaired in intact cells depleted of imp7, and DNA complexes remained localized in the cytoplasm. Poor DNA nuclear import in cells depleted of imp7 directly correlated with lower gene expression levels in these cells compared to controls. Inefficient nuclear import of transfected DNA induced greater upregulation of the interferon pathway, suggesting that rapid DNA nuclear import may prevent uncontrolled activation of the innate immune response. Our results provide evidence that imp7 is a non-redundant component of an intrinsic pathway in mammalian cells for efficient accumulation of exogenous and endogenous DNA in the nucleus, which may be critical for the exchange of genetic information between mitochondria and nuclear genomes and to control activation of the innate immune response. PMID:23067392

  4. Modification of hot cells for general purpose heat source assembly at the radioisotope power systems facility

    NASA Astrophysics Data System (ADS)

    Carteret, Betty A.

    1992-01-01

    Eight existing, unused hot cells currently are being modified for use in the Radioisotope Power Systems Facility (RPSF) to assemble 238Pu-fueled heat sources for Radioisotope Thermoelectric Generators (RTGs). Four air atmosphere cells will be used for storage, decanning, and decontamination of the iridium-clad radioisotope fuel. The remaining four argon atmosphere cells will be used to assemble fuel and graphite components for production and packaging of general purpose heat source (GPHS) assembly modules, which provide heat to drive the thermoelectric conversion process in the generators. The hot cells will be equipped to perform remote and glovebox-type operations. They will provide shielding and contamination control measures to reduce worker radiation exposure to levels within current U.S. Department of Energy (DOE) guidelines. Designs emphasize the Westinghouse Hanford Company (Westinghouse Hanford) as low as reasonably achievable (ALARA) radiation protection policy.

  5. Modification of hot cells for general purpose heat source assembly at the Radioisotope Power Systems Facility

    NASA Astrophysics Data System (ADS)

    Carteret, B. A.

    1991-09-01

    Eight existing, unused hot cells currently are being modified for use in the Radioisotope Power Systems Facility (RPSF) to assemble Pu-238 fueled heat sources for radioisotope thermoelectric generators (RTGs). Four air atmosphere cells will be used for storage, decanning, and decontamination of the iridium-clad radioisotope fuel. The remaining four argon atmosphere cells will be used to assemble fuel and graphite components for production and packaging of general purpose heat source (GPHS) assembly modules, which provide heat to drive the thermoelectric conversion process in the generators. The hot cells will be equipped to perform remote and glovebox-type operations. They will provide shielding and contamination control measures to reduce worker radiation exposure to levels within current U.S. Department of Energy (DOE) guidelines. Designs emphasize the Westinghouse Hanford Company (Westinghouse Hanford) as low as reasonably achievable (ALARA) radiation protection policy.

  6. The Nuclear Option: Evidence Implicating the Cell Nucleus in Mechanotransduction.

    PubMed

    Szczesny, Spencer E; Mauck, Robert L

    2017-02-01

    Biophysical stimuli presented to cells via microenvironmental properties (e.g., alignment and stiffness) or external forces have a significant impact on cell function and behavior. Recently, the cell nucleus has been identified as a mechanosensitive organelle that contributes to the perception and response to mechanical stimuli. However, the specific mechanotransduction mechanisms that mediate these effects have not been clearly established. Here, we offer a comprehensive review of the evidence supporting (and refuting) three hypothetical nuclear mechanotransduction mechanisms: physical reorganization of chromatin, signaling at the nuclear envelope, and altered cytoskeletal structure/tension due to nuclear remodeling. Our goal is to provide a reference detailing the progress that has been made and the areas that still require investigation regarding the role of nuclear mechanotransduction in cell biology. Additionally, we will briefly discuss the role that mathematical models of cell mechanics can play in testing these hypotheses and in elucidating how biophysical stimulation of the nucleus drives changes in cell behavior. While force-induced alterations in signaling pathways involving lamina-associated polypeptides (LAPs) (e.g., emerin and histone deacetylase 3 (HDAC3)) and transcription factors (TFs) located at the nuclear envelope currently appear to be the most clearly supported mechanism of nuclear mechanotransduction, additional work is required to examine this process in detail and to more fully test alternative mechanisms. The combination of sophisticated experimental techniques and advanced mathematical models is necessary to enhance our understanding of the role of the nucleus in the mechanotransduction processes driving numerous critical cell functions.

  7. Proton nuclear magnetic resonance studies on bulge-containing DNA oligonucleotides from a mutational hot-spot sequence

    SciTech Connect

    Woodson, S.A.; Crothers, D.M.

    1987-02-10

    A series of bulge-containing and normal double-helical synthetic oligodeoxyribonucleotides, of sequence corresponding to a frame-shift mutational hot spot in the lambda C/sub I/ gene, are compared by proton magnetic resonance spectroscopy at 500 MHz. The imino proton resonances are assigned by one-dimensional nuclear Overhauser effect spectroscopy. Nonselective T/sub 1/ inversion-recovery experiments are used to determine exchangeable proton lifetimes and to compare helix stability and dynamics of the three duplexes. An extra adenosine flanking the internal G-C base pairs has a strongly localized effect on helix stability, but the destabilizing effect of an extra cytidine in a C tract is delocalized over the entire G-C run. These data lead to the conclusion that the position of the bulge migrates along the run in the fast-exchange limit on the NMR time scale. Rapid migration of the bulge defect in homopolymeric sequences may help rationalize both frame-shift mutagenesis and translational frame shifting. The authors estimate that the unfavorable free energy of a localized bulge defect is 2.9-3.2 kcal/mol, in good agreement with earlier estimates for RNA helices.

  8. Dynamical ejecta from precessing neutron star-black hole mergers with a hot, nuclear-theory based equation of state

    NASA Astrophysics Data System (ADS)

    Foucart, F.; Desai, D.; Brege, W.; Duez, M. D.; Kasen, D.; Hemberger, D. A.; Kidder, L. E.; Pfeiffer, H. P.; Scheel, M. A.

    2017-02-01

    Neutron star-black hole binaries are among the strongest sources of gravitational waves detectable by current observatories. They can also power bright electromagnetic signals (gamma-ray bursts, kilonovae), and may be a significant source of production of r-process nuclei. A misalignment of the black hole spin with respect to the orbital angular momentum leads to precession of that spin and of the orbital plane, and has a significant effect on the properties of the post-merger remnant and of the material ejected by the merger. We present a first set of simulations of precessing neutron star-black hole mergers using a hot, composition dependent, nuclear-theory based equation of state (DD2). We show that the mass of the remnant and of the dynamical ejecta are broadly consistent with the result of simulations using simpler equations of state, while differences arise when considering the dynamics of the merger and the velocity of the ejecta. We show that the latter can easily be understood from assumptions about the composition of low-density, cold material in the different equations of state, and propose an updated estimate for the ejecta velocity which takes those effects into account. We also present an updated mesh-refinement algorithm which allows us to improve the numerical resolution used to evolve neutron star-black hole mergers.

  9. A&M. TAN633. Sections show view of hot cell caskentry doors, ...

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

    A&M. TAN-633. Sections show view of hot cell cask-entry doors, manipulators in each cell, drainage trenches, door and room details. Ralph M. Parsons 1229-13-ANP/GE-3-633-A-2. Date: December 1956. Approved by INEEL Classification Office for public release. INNEL index code no. 034-0633-00-693-107316 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  10. The impact of hot charge carrier mobility on photocurrent losses in polymer-based solar cells.

    PubMed

    Philippa, Bronson; Stolterfoht, Martin; Burn, Paul L; Juška, Gytis; Meredith, Paul; White, Ronald D; Pivrikas, Almantas

    2014-07-22

    A typical signature of charge extraction in disordered organic systems is dispersive transport, which implies a distribution of charge carrier mobilities that negatively impact on device performance. Dispersive transport has been commonly understood to originate from a time-dependent mobility of hot charge carriers that reduces as excess energy is lost during relaxation in the density of states. In contrast, we show via photon energy, electric field and film thickness independence of carrier mobilities that the dispersive photocurrent in organic solar cells originates not from the loss of excess energy during hot carrier thermalization, but rather from the loss of carrier density to trap states during transport. Our results emphasize that further efforts should be directed to minimizing the density of trap states, rather than controlling energetic relaxation of hot carriers within the density of states.

  11. New electron beam facility for irradiated plasma facing materials testing in hot cell

    SciTech Connect

    Sakamoto, N.; Kawamura, H.; Akiba, M.

    1995-09-01

    Since plasma facing components such as the first wall and the divertor for the next step fusion reactors are exposed to high heat loads and high energy neutron flux generated by the plasma, it is urgent to develop of plasma facing components which can resist these. Then, we have established electron beam heat facility ({open_quotes}OHBIS{close_quotes}, Oarai Hot-cell electron Beam Irradiating System) at a hot cell in JMTR (Japan Materials Testing Reactor) hot laboratory in order to estimate thermal shock resistivity of plasma facing materials and heat removal capabilities of divertor elements under steady state heating. In this facility, irradiated plasma facing materials (beryllium, carbon based materials and so on) and divertor elements can be treated. This facility consists of an electron beam unit with the maximum beam power of 50kW and the vacuum vessel. The acceleration voltage and the maximum beam current are 30kV (constant) and 1.7A, respectively. The loading time of electron beam is more than 0.1ms. The shape of vacuum vessel is cylindrical, and the mainly dimensions are 500mm in inner diameter, 1000mm in height. The ultimate vacuum of this vessel is 1 x 10{sup -4}Pa. At present, the facility for thermal shock test has been established in a hot cell. And performance estimation on the electron beam is being conducted. Presently, the devices for heat loading tests under steady state will be added to this facility.

  12. Building Electromagnetic Hot Spots in Living Cells via Target-Triggered Nanoparticle Dimerization.

    PubMed

    Zhou, Wen; Li, Qiang; Liu, Huiqiao; Yang, Jie; Liu, Dingbin

    2017-03-10

    Electromagnetic hot spots of surface-enhanced Raman scattering have been extensively employed for bioanalysis in solution or on a substrate, but building hot spots in living systems for probing targets of interest has not been achieved yet because of the complex and dynamic physiological environment. Herein, we show that a target-programmed nanoparticle dimerization can be combined with the background-free Raman reporters (alkyne, C≡C; nitrile, C≡N) for multiplexed imaging of microRNAs (miRNAs) in living cells. The in situ formation of plasmonic dimers results in an intense hot spot, thus dramatically enhancing the Raman signals of the reporters residing in the hot spot. More significantly, the reporters exhibit single nonoverlapping peaks in the cellular Raman-silent region (1800-2800 cm(-1)), thus eliminating spectral unmixing and background interference. A 3D Raman mapping technique was harnessed to monitor the spatial distribution of the dimers and thus the multiple miRNAs in cells. This approach could be extended to probe other biomarkers of interest for monitoring specific pathophysiological events at the live-cell level.

  13. Hot Start to European Pluripotent Stem Cell Banking.

    PubMed

    De Sousa, Paul A; Steeg, Rachel; Kreisel, Beate; Allsopp, Timothy E

    2017-07-01

    Achieving consistency in standards of access to and quality of human induced pluripotent stem cells has lagged behind their use. In Europe, a network of academic and industrial partners has been established to overcome this challenge. The experience reveals the devil in the detail of worthy ambitions informing future efforts. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Joint modeling of cell and nuclear shape variation

    PubMed Central

    Johnson, Gregory R.; Buck, Taraz E.; Sullivan, Devin P.; Rohde, Gustavo K.; Murphy, Robert F.

    2015-01-01

    Modeling cell shape variation is critical to our understanding of cell biology. Previous work has demonstrated the utility of nonrigid image registration methods for the construction of nonparametric nuclear shape models in which pairwise deformation distances are measured between all shapes and are embedded into a low-dimensional shape space. Using these methods, we explore the relationship between cell shape and nuclear shape. We find that these are frequently dependent on each other and use this as the motivation for the development of combined cell and nuclear shape space models, extending nonparametric cell representations to multiple-component three-dimensional cellular shapes and identifying modes of joint shape variation. We learn a first-order dynamics model to predict cell and nuclear shapes, given shapes at a previous time point. We use this to determine the effects of endogenous protein tags or drugs on the shape dynamics of cell lines and show that tagged C1QBP reduces the correlation between cell and nuclear shape. To reduce the computational cost of learning these models, we demonstrate the ability to reconstruct shape spaces using a fraction of computed pairwise distances. The open-source tools provide a powerful basis for future studies of the molecular basis of cell organization. PMID:26354424

  15. Study of LO-phonon decay in semiconductors for hot carrier solar cell

    NASA Astrophysics Data System (ADS)

    Levard, Hugo; Vidal, Julien; Laribi, Sana; Guillemoles, Jean-François

    2014-03-01

    Knowledge of phonon decay is of crucial importance when studying basic properties of semiconductors, since they are closely related to Raman linewidth and non-equilibrium-hot-carriers cooling. The latter indeed cools down to the bottom of the conduction band within a picosecond range because of electron-phonon interaction. The eventual emitted hot phonons then decay in few picoseconds. The hot carriers cooling can be slowed down by considering the decay rate dependence of phonon on conservation rules, whose tuning may reduce the allowed two-phonon final states density. This is of direct interest for the third generation photovoltaic devices that are Hot Carrier Solar Cells (HCSC), in which the photoexcited carriers are extracted at an energy higher than thermal equilibrium. One of the HCSC main challenges then is to find an absorber material in which the hot phonons has a relaxation time longer than the carriers cooling time, so that we can expect the electron to ``reabsorb'' a phonon, slowing down the electronic cooling. HCSC yield is ultimately limited by LO phonon decay, though. In this work, we present theoretical results obtained from ab initio calculations of phonon lifetime in III-V and IV-IV semiconductors through a three-phonon process. Common approximations in the literature are questioned. In particular, we show that the usual ``zone-center approximation'' is not valid in some specific semiconductors. The analysis allows to correctly investigate phonon decay mechanisms in bulk and nanostructured materials.

  16. Cloning mice and ES cells by nuclear transfer from somatic stem cells and fully differentiated cells.

    PubMed

    Wang, Zhongde

    2011-01-01

    Cloning animals by nuclear transfer (NT) has been successful in several mammalian species. In addition to cloning live animals (reproductive cloning), this technique has also been used in several species to establish cloned embryonic stem (ntES) cell lines from somatic cells. It is the latter application of this technique that has been heralded as being the potential means to produce isogenic embryonic stem cells from patients for cell therapy (therapeutic cloning). These two types of cloning differ only in the steps after cloned embryos are produced: for reproductive cloning the cloned embryos are transferred to surrogate mothers to allow them to develop to full term and for therapeutic cloning the cloned embryos are used to derive ntES cells. In this chapter, a detailed NT protocol in mouse by using somatic stem cells (neuron and skin stem cells) and fully differentiated somatic cells (cumulus cells and fibroblast cells) as nuclear donors is described.

  17. Nuclear envelope rupture and repair during cancer cell migration

    PubMed Central

    Denais, Celine M.; Gilbert, Rachel M.; Isermann, Philipp; McGregor, Alexandra L.; te Lindert, Mariska; Weigelin, Bettina; Davidson, Patricia M.; Friedl, Peter; Wolf, Katarina; Lammerding, Jan

    2016-01-01

    During cancer metastasis, tumor cells penetrate tissues through tight interstitial spaces, requiring extensive deformation of the cell and its nucleus. Here, we investigated tumor cell migration in confining microenvironments in vitro and in vivo. Nuclear deformation caused localized loss of nuclear envelope (NE) integrity, which led to the uncontrolled exchange of nucleo-cytoplasmic content, herniation of chromatin across the NE, and DNA damage. The incidence of NE rupture increased with cell confinement and with depletion of nuclear lamins, NE proteins that structurally support the nucleus. Cells restored NE integrity using components of the endosomal sorting complexes required for transport-III (ESCRT-III) machinery. Our findings indicate that cell migration incurs substantial physical stress on the NE and its content, requiring efficient NE and DNA damage repair for survival. PMID:27013428

  18. The feasibility study of hot cell decontamination by the PFC spray method

    SciTech Connect

    Hui-Jun Won; Chong-Hun Jung; Jei-Kwon Moon

    2008-01-15

    module. A performance test on each module was executed and the results have been reported. A combined test of the four modules, however, has not been performed as yet. The main objective of the present study is to demonstrate the feasibility of the full PFC spray decontamination process. Decontamination of the inside of the IMEF hot cell by the PFC spray method was also performed. PFC spray decontamination process was demonstrated by using a surrogate wall contaminated with Eu{sub 2}O{sub 3} powder. The spray pressure was 41 kgf/cm{sup 2}, the orifice diameter was 0.2 mm and the spray velocity was 0.2 L/min. And, the decontaminated area was 100 cm{sup 2}. From previous test results, we found that the decontamination factor of the PFC spray method was in the range from 9.6 to 62.4. When the decontamination efficiency of Co-60 was high, then the decontamination efficiency of Cs-137 was also high. As the surface roughness of the specimen increased, the PFC spray decontamination efficiency decreased. Inferring from the previous results, the surface of the surrogate wall was cleaned by the PFC spray method. The vacuum cup of the collection module operated well and gathered more than 99 % of the PFC solution. Also, filtration and distillation modules operated well. All the filtered PFC solution flowed to the storage chamber where some of the PFC solution was distilled. The coolant of the distillation module was a dry ice. And, the recycled solution was transferred to the spray module by a high pressure pump. To evaluate the PFC spray decontamination efficiency, a smear device was fabricated and operated by a manipulator. Before and after decontamination, a smear test was performed. The tested area was 100 cm{sup 2} and the radioactivity was estimated indirectly by measuring the radioactivity of the filter paper. The average decontamination factor was in the range between 10 and 15. One application time was 2 minutes. The sprayed PFC solution was collected by the vacuum cup and

  19. Transient nuclear envelope rupturing during interphase in human cancer cells

    PubMed Central

    Vargas, Jesse D.; Hatch, Emily M.; Anderson, Daniel J.; Hetzer, Martin W.

    2012-01-01

    Neoplastic cells are often characterized by specific morphological abnormalities of the nuclear envelope (NE), which have been used for cancer diagnosis for more than a century. The NE is a double phospholipid bilayer that encapsulates the nuclear genome, regulates all nuclear trafficking of RNAs and proteins and prevents the passive diffusion of macromolecules between the nucleoplasm and the cytoplasm. Whether there is a consequence to the proper functioning of the cell and loss of structural integrity of the nucleus remains unclear. Using live cell imaging, we characterize a phenomenon wherein nuclei of several proliferating human cancer cell lines become temporarily ruptured during interphase. Strikingly, NE rupturing was associated with the mislocalization of nucleoplasmic and cytoplasmic proteins and, in the most extreme cases, the entrapment of cytoplasmic organelles in the nuclear interior. In addition, we observed the formation of micronuclei-like structures during interphase and the movement of chromatin out of the nuclear space. The frequency of these NE rupturing events was higher in cells in which the nuclear lamina, a network of intermediate filaments providing mechanical support to the NE, was not properly formed. Our data uncover the existence of a NE instability that has the potential to change the genomic landscape of cancer cells. PMID:22567193

  20. Regulation of proliferating cell nuclear antigen ubiquitination in mammalian cells

    PubMed Central

    Niimi, Atsuko; Brown, Stephanie; Sabbioneda, Simone; Kannouche, Patricia L.; Scott, Andrew; Yasui, Akira; Green, Catherine M.; Lehmann, Alan R.

    2008-01-01

    After exposure to DNA-damaging agents that block the progress of the replication fork, monoubiquitination of proliferating cell nuclear antigen (PCNA) mediates the switch from replicative to translesion synthesis DNA polymerases. We show that in human cells, PCNA is monoubiquitinated in response to methyl methanesulfonate and mitomycin C, as well as UV light, albeit with different kinetics, but not in response to bleomycin or camptothecin. Cyclobutane pyrimidine dimers are responsible for most of the PCNA ubiquitination events after UV-irradiation. Failure to ubiquitinate PCNA results in substantial sensitivity to UV and methyl methanesulfonate, but not to camptothecin or bleomycin. PCNA ubiquitination depends on Replication Protein A (RPA), but is independent of ATR-mediated checkpoint activation. After UV-irradiation, there is a temporal correlation between the disappearance of the deubiquitinating enzyme USP1 and the presence of PCNA ubiquitination, but this correlation was not found after chemical mutagen treatment. By using cells expressing photolyases, we are able to remove the UV lesions, and we show that PCNA ubiquitination persists for many hours after the damage has been removed. We present a model of translesion synthesis behind the replication fork to explain the persistence of ubiquitinated PCNA. PMID:18845679

  1. 324 and 325 Building Hot Cell Cleanout Program: Air lock cover block refurbishment

    SciTech Connect

    Katayama, Y.B.; Holton, L.K. Jr.; Gale, R.M.

    1989-05-01

    The high-density concrete cover blocks shielding the pipe trench in the hot-cell air lock of the 324 Building Radiochemical Engineering Cells had accumulated fixed radioactivity ranging from 1100 to 22, 000 mrad/hr. A corresponding increase in the radiation exposure to personnel entering the air lock, together with ALARA concerns, led to the removal of the contaminated concrete surface with a hydraulic spaller and the emplacement of a stainless steel covering over a layer of grout. The resultant saving in radiation exposure is estimated to be 7200 mrad for personnel completing burial box runs for the 324 and 325 Building Hot Cell Cleanout Program. Radiation exposure to all staff members entering the air lock is now at least 50% lower. 3 refs., 22 figs., 1 tab.

  2. A simple polymeric model describes cell nuclear mechanical response

    NASA Astrophysics Data System (ADS)

    Banigan, Edward; Stephens, Andrew; Marko, John

    The cell nucleus must continually resist inter- and intracellular mechanical forces, and proper mechanical response is essential to basic cell biological functions as diverse as migration, differentiation, and gene regulation. Experiments probing nuclear mechanics reveal that the nucleus stiffens under strain, leading to two characteristic regimes of force response. This behavior depends sensitively on the intermediate filament protein lamin A, which comprises the outer layer of the nucleus, and the properties of the chromatin interior. To understand these mechanics, we study a simulation model of a polymeric shell encapsulating a semiflexible polymer. This minimalistic model qualitatively captures the typical experimental nuclear force-extension relation and observed nuclear morphologies. Using a Flory-like theory, we explain the simulation results and mathematically estimate the force-extension relation. The model and experiments suggest that chromatin organization is a dominant contributor to nuclear mechanics, while the lamina protects cell nuclei from large deformations.

  3. Nanotopographical Modulation of Cell Function through Nuclear Deformation

    PubMed Central

    Wang, Kai; Bruce, Allison; Mezan, Ryan; Kadiyala, Anand; Wang, Liying; Dawson, Jeremy; Rojanasakul, Yon; Yang, Yong

    2016-01-01

    Although nanotopography has been shown to be a potent modulator of cell behavior, it is unclear how the nanotopographical cue, through focal adhesions, affects the nucleus, eventually influencing cell phenotype and function. Thus, current methods to apply nanotopography to regulate cell behavior are basically empirical. We, herein, engineered nanotopographies of various shapes (gratings and pillars) and dimensions (feature size, spacing and height), and thoroughly investigated cell spreading, focal adhesion organization and nuclear deformation of human primary fibroblasts as the model cell grown on the nanotopographies. We examined the correlation between nuclear deformation and cell functions such as cell proliferation, transfection and extracellular matrix protein type I collagen production. It was found that the nanoscale gratings and pillars could facilitate focal adhesion elongation by providing anchoring sites, and the nanogratings could orient focal adhesions and nuclei along the nanograting direction, depending on not only the feature size but also the spacing of the nanogratings. Compared with continuous nanogratings, discrete nanopillars tended to disrupt the formation and growth of focal adhesions and thus had less profound effects on nuclear deformation. Notably, nuclear volume could be effectively modulated by the height of nanotopography. Further, we demonstrated that cell proliferation, transfection, and type I collagen production were strongly associated with the nuclear volume, indicating that the nucleus serves as a critical mechanosensor for cell regulation. Our study delineated the relationships between focal adhesions, nucleus and cell function and highlighted that the nanotopography could regulate cell phenotype and function by modulating nuclear deformation. This study provides insight into the rational design of nanotopography for new biomaterials and the cell–substrate interfaces of implants and medical devices. PMID:26844365

  4. [Nuclear transfer of goat somatic cells transgenic for human lactoferrin].

    PubMed

    Li, Lan; Shen, Wei; Pan, Qing-Yu; Min, Ling-Jiang; Sun, Yu-Jiang; Fang, Yong-Wei; Deng, Ji-Xian; Pan, Qing-Jie

    2006-12-01

    Transgenic animal mammary gland bioreactors are being used to produce recombinant proteins with appropriate post-translational modifications, and nuclear transfer of transgenic somatic cells is a more powerful method to produce mammary gland bioreactor. Here we describe efficient gene transfer and nuclear transfer in goat somatic cells. Gene targeting vector pGBC2LF was constructed by cloning human lactoferrin (LF) gene cDNA into exon 2 of the milk goat beta-casein gene, and the endogenous start condon was replaced by that of human LF gene. Goat fetal fibroblasts were transfected with linearized pGBC2LF and 14 cell lines were positive according to PCR and Southern blot. The transgenic cells were used as donor cells of nuclear transfer, and some of reconstructed embryos could develop to blastocyst in vitro.

  5. The Mammalian Cell Cycle Regulates Parvovirus Nuclear Capsid Assembly

    PubMed Central

    Riolobos, Laura; Domínguez, Carlos; Kann, Michael; Almendral, José M.

    2015-01-01

    It is unknown whether the mammalian cell cycle could impact the assembly of viruses maturing in the nucleus. We addressed this question using MVM, a reference member of the icosahedral ssDNA nuclear parvoviruses, which requires cell proliferation to infect by mechanisms partly understood. Constitutively expressed MVM capsid subunits (VPs) accumulated in the cytoplasm of mouse and human fibroblasts synchronized at G0, G1, and G1/S transition. Upon arrest release, VPs translocated to the nucleus as cells entered S phase, at efficiencies relying on cell origin and arrest method, and immediately assembled into capsids. In synchronously infected cells, the consecutive virus life cycle steps (gene expression, proteins nuclear translocation, capsid assembly, genome replication and encapsidation) proceeded tightly coupled to cell cycle progression from G0/G1 through S into G2 phase. However, a DNA synthesis stress caused by thymidine irreversibly disrupted virus life cycle, as VPs became increasingly retained in the cytoplasm hours post-stress, forming empty capsids in mouse fibroblasts, thereby impairing encapsidation of the nuclear viral DNA replicative intermediates. Synchronously infected cells subjected to density-arrest signals while traversing early S phase also blocked VPs transport, resulting in a similar misplaced cytoplasmic capsid assembly in mouse fibroblasts. In contrast, thymidine and density arrest signals deregulating virus assembly neither perturbed nuclear translocation of the NS1 protein nor viral genome replication occurring under S/G2 cycle arrest. An underlying mechanism of cell cycle control was identified in the nuclear translocation of phosphorylated VPs trimeric assembly intermediates, which accessed a non-conserved route distinct from the importin α2/β1 and transportin pathways. The exquisite cell cycle-dependence of parvovirus nuclear capsid assembly conforms a novel paradigm of time and functional coupling between cellular and virus life

  6. Hot-Wire CVD Amorphous Si Materials for Solar Cell Application

    SciTech Connect

    Wang, Q.

    2009-01-01

    Hydrogenated amorphous silicon (a-Si:H) thin films and their application to solar cells fabricated using the hot-wire chemical vapor deposition (HWCVD) or (CAT)-CVD will be reviewed. This review will focus on the comparison to the standard plasma enhance (PE) CVD in the terms of deposition technique, film properties, and solar cell performance. The advantages of using HWCVD for a-Si:H solar cell research as well as the criteria for industry's adaptation of this technique for mass production will be addressed.

  7. Cell fusion through a microslit between adhered cells and observation of their nuclear behavior.

    PubMed

    Wada, Ken-Ichi; Hosokawa, Kazuo; Kondo, Eitaro; Ito, Yoshihiro; Maeda, Mizuo

    2014-07-01

    This paper describes a novel cell fusion method which induces cell fusion between adhered cells through a microslit for preventing nuclear mixing. For this purpose, a microfluidic device which had ∼ 100 cell pairing structures (CPSs) making cell pairs through microslits with 2.1 ± 0.3 µm width was fabricated. After trapping NIH3T3 cells with hydrodynamic forces at the CPSs, the cells were fused through the microslit by the Sendai virus envelope method. With following timelapse observation, we discovered that the spread cells were much less susceptible to nuclear migration passing through the microslit compared with round cells, and that cytoplasmic fraction containing mitochondria was transferred through the microslit without nuclear mixing. These findings will provide an effective method for cell fusion without nuclear mixing, and will lead to an efficient method for reprograming and transdifferentiation of target cells toward regenerative medicine. © 2014 Wiley Periodicals, Inc.

  8. Nuclear microscopy of sperm cell elemental structure

    SciTech Connect

    Bench, G.S.; Balhorn, R.; Friz, A.M.; Freeman, S.P.H.T.

    1994-09-28

    Theories suggest there is a link between protamine concentrations in individual sperm and male fertility. Previously, biochemical analyses have used pooled samples containing millions of sperm to determine protamine concentrations. These methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. Nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the amount of phosphorus and sulfur, the total DNA and protamine content in individual sperm from fertile bull and mouse semen have been determined. These values agree with results obtained from other biochemical analyses. Nuclear microscopy shows promise for measuring elemental profiles in the chromatin of individual sperm. The technique may be able to resolve theories regarding the importance of protamines to male fertility and identify biochemical defects responsible for certain types of male infertility.

  9. Nuclear microscopy of sperm cell elemental structure

    NASA Astrophysics Data System (ADS)

    Bench, Graham S.; Balhorn, Rod; Friz, Alexander M.

    1995-05-01

    Theories suggest there is a link between protamine concentrations in individual sperm and male fertility. Previously, biochemical analyses have used pooled samples containing millions of sperm to determine protamine concentrations. These methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. Nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the amount of phosphorus and sulfur, the total DNA and protamine content in individual sperm from fertile bull and mouse semen have been determined. These values agree with results obtained from other biochemical analyses. Nuclear microscopy shows promise for measuring elemental profiles in the chromatin of individual sperm. The technique may be able to resolve theories regarding the importance of protamines to male fertility and identify biochemical defects responsible for certain types of male infertility.

  10. Galvanic cell for processing of used nuclear fuel

    DOEpatents

    Garcia-Diaz, Brenda L.; Martinez-Rodriguez, Michael J.; Gray, Joshua R.; Olson, Luke C.

    2017-02-07

    A galvanic cell and methods of using the galvanic cell is described for the recovery of uranium from used nuclear fuel according to an electrofluorination process. The galvanic cell requires no input energy and can utilize relatively benign gaseous fluorinating agents. Uranium can be recovered from used nuclear fuel in the form of gaseous uranium compound such as uranium hexafluoride, which can then be converted to metallic uranium or UO.sub.2 and processed according to known methodology to form a useful product, e.g., fuel pellets for use in a commercial energy production system.

  11. Generation of bovine transgenics using somatic cell nuclear transfer

    PubMed Central

    Hodges, Craig A; Stice, Steven L

    2003-01-01

    The ability to produce transgenic animals through the introduction of exogenous DNA has existed for many years. However, past methods available to generate transgenic animals, such as pronuclear microinjection or the use of embryonic stem cells, have either been inefficient or not available in all animals, bovine included. More recently somatic cell nuclear transfer has provided a method to create transgenic animals that overcomes many deficiencies present in other methods. This review summarizes the benefits of using somatic cell nuclear transfer to create bovine transgenics as well as the possible opportunities this method creates for the future. PMID:14613543

  12. Generation of bovine transgenics using somatic cell nuclear transfer.

    PubMed

    Hodges, Craig A; Stice, Steven L

    2003-11-07

    The ability to produce transgenic animals through the introduction of exogenous DNA has existed for many years. However, past methods available to generate transgenic animals, such as pronuclear microinjection or the use of embryonic stem cells, have either been inefficient or not available in all animals, bovine included. More recently somatic cell nuclear transfer has provided a method to create transgenic animals that overcomes many deficiencies present in other methods. This review summarizes the benefits of using somatic cell nuclear transfer to create bovine transgenics as well as the possible opportunities this method creates for the future.

  13. Cloning animals by somatic cell nuclear transfer – biological factors

    PubMed Central

    Tian, X Cindy; Kubota, Chikara; Enright, Brian; Yang, Xiangzhong

    2003-01-01

    Cloning by nuclear transfer using mammalian somatic cells has enormous potential application. However, somatic cloning has been inefficient in all species in which live clones have been produced. High abortion and fetal mortality rates are commonly observed. These developmental defects have been attributed to incomplete reprogramming of the somatic nuclei by the cloning process. Various strategies have been used to improve the efficiency of nuclear transfer, however, significant breakthroughs are yet to happen. In this review we will discuss studies conducted, in our laboratories and those of others, to gain a better understanding of nuclear reprogramming. Because cattle are a species widely used for nuclear transfer studies, and more laboratories have succeeded in cloning cattle than any other specie, this review will be focused on somatic cell cloning of cattle. PMID:14614770

  14. A Change In Nuclear Pore Complex Composition Regulates Cell Differentiation

    PubMed Central

    D’Angelo, Maximiliano A.; Gomez-Cavazos, J. Sebastian; Mei, Arianna; Lackner, Daniel H.; Hetzer, Martin W.

    2011-01-01

    SUMMARY Nuclear pore complexes (NPCs) are built from ~30 different proteins called nucleoporins. Previous studies have shown that several Nups exhibit cell-type-specific expression and that mutations in NPC components result in tissue-specific diseases. Here we show that a specific change in NPC composition is required for both myogenic and neuronal differentiation. The transmembrane nucleoporin Nup210 is absent in proliferating myoblasts and embryonic stem (ES) cells but becomes expressed and incorporated into NPCs during cell differentiation. Preventing Nup210 production by RNAi blocks myogenesis and the differentiation of ES cells into neuroprogenitors. We found that the addition of Nup210 to NPCs does not affect nuclear transport but is required for the induction of genes that are essential for cell differentiation. Our results identify a single change in NPC composition as an essential step in cell differentiation and establish a role for Nup210 in gene expression regulation and cell fate determination. PMID:22264802

  15. HOT CELL BUILDING, TRA632, INTERIOR. WINDOWED ROOM IS OFFICE; NEXT ...

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

    HOT CELL BUILDING, TRA-632, INTERIOR. WINDOWED ROOM IS OFFICE; NEXT DOOR WAS DARKROOM, AND THIRD DOOR LED TO ANOTHER OFFICE. ALL ARE ALONG NORTH WALL OF BUILDING (ETR EXTENSION OF 1958). CAMERA FACES NORTHEAST. PUMICE BLOCK WALLS. INL NEGATIVE NO. HD46-29-1. Mike Crane, Photographer, 2/2005 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  16. A&M. TAN633. Hot cell floor plans, elevations, sections. Hole schedule ...

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

    A&M. TAN-633. Hot cell floor plans, elevations, sections. Hole schedule (penetrations through concrete). Swing-door details. Ralph M. Parsons 1229-13-ANP/GE-3-633-A-3. Date: December 1956. Approved by INEEL Classification Office for public release. INNEL index code no. 034-0633-00-693-107317 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

  17. Nuclear tristetraprolin acts as a corepressor of multiple steroid nuclear receptors in breast cancer cells.

    PubMed

    Barrios-García, Tonatiuh; Gómez-Romero, Vania; Tecalco-Cruz, Ángeles; Valadéz-Graham, Viviana; León-Del-Río, Alfonso

    2016-06-01

    Tristetraprolin (TTP) is a 34-kDa, zinc finger-containing factor that in mammalian cells acts as a tumor suppressor protein through two different mechanisms. In the cytoplasm TTP promotes the decay of hundreds of mRNAs encoding cell factors involved in inflammation, tissue invasion, and metastasis. In the cell nucleus TTP has been identified as a transcriptional corepressor of the estrogen receptor alpha (ERα), which has been associated to the development and progression of the majority of breast cancer tumors. In this work we report that nuclear TTP modulates the transactivation activity of progesterone receptor (PR), glucocorticoid receptor (GR) and androgen receptor (AR). In recent years these steroid nuclear receptors have been shown to be of clinical and therapeutical relevance in breast cancer. The functional association between TTP and steroid nuclear receptors is supported by the finding that TTP physically interacts with ERα, PR, GR and AR in vivo. We also show that TTP overexpression attenuates the transactivation of all the steroid nuclear receptors tested. In contrast, siRNA-mediated reduction of endogenous TTP expression in MCF-7 cells produced an increase in the transcriptional activities of ERα, PR, GR and AR. Taken together, these results suggest that the function of nuclear TTP in breast cancer cells is to act as a corepressor of ERα, PR, GR and AR. We propose that the reduction of TTP expression observed in different types of breast cancer tumors may contribute to the development of this disease by producing a dysregulation of the transactivation activity of multiple steroid nuclear receptors.

  18. Inside the Cell: Integrins as New Governors of Nuclear Alterations?

    PubMed Central

    Madrazo, Elena; Cordero Conde, Andrea

    2017-01-01

    Cancer cell migration is a complex process that requires coordinated structural changes and signals in multiple cellular compartments. The nucleus is the biggest and stiffest organelle of the cell and might alter its physical properties to allow cancer cell movement. Integrins are transmembrane receptors that mediate cell-cell and cell-extracellular matrix interactions, which regulate numerous intracellular signals and biological functions under physiological conditions. Moreover, integrins orchestrate changes in tumor cells and their microenvironment that lead to cancer growth, survival and invasiveness. Most of the research efforts have focused on targeting integrin-mediated adhesion and signaling. Recent exciting data suggest the crucial role of integrins in controlling internal cellular structures and nuclear alterations during cancer cell migration. Here we review the emerging role of integrins in nuclear biology. We highlight increasing evidence that integrins are critical for changes in multiple nuclear components, the positioning of the nucleus and its mechanical properties during cancer cell migration. Finally, we discuss how integrins are integral proteins linking the plasma membrane and the nucleus, and how they control cell migration to enable cancer invasion and infiltration. The functional connections between these cell receptors and the nucleus will serve to define new attractive therapeutic targets. PMID:28684679

  19. Evaluation of Tritium Behavior in the Epoxy Painted Concrete Wall of ITER Hot Cell

    SciTech Connect

    Nakamura, Hirofumi; Hayashi, Takumi; Kobayashi, Kazuhiro; Nishi, Masataka

    2005-07-15

    Tritium behavior released in the ITER hot cell has been investigated numerically using a combined analytical methods of a tritium transport analysis in the multi-layer wall (concrete and epoxy paint) with the one dimensional diffusion model and a tritium concentration analysis in the hot cell with the complete mixing model by the ventilation. As the results, it is revealed that tritium concentration decay and permeation issues are not serious problem in a viewpoint of safety, since it is expected that tritium concentration in the hot cell decrease rapidly within several days just after removing the tritium release source, and tritium permeation through the epoxy painted concrete wall will be negligible as long as the averaged realistic diffusion coefficient is ensured in the concrete wall. It is also revealed that the epoxy paint on the concrete wall prevents the tritium inventory increase in the concrete wall greatly (two orders of magnitudes), but still, the inventory in the wall is estimated to reach about 0.1 PBq for 20 years operation.

  20. Efficacy of disinfectants and hot water against biofilm cells of Burkholderia cepacia.

    PubMed

    Miyano, Naoyuki; Oie, Shigeharu; Kamiya, Akira

    2003-05-01

    The effects of various disinfectants and hot water on planktonic cells and biofilm cells of Burkholderia cepacia were investigated. The survival rate of viable B. cepacia cells in suspension decreased to 0.001% or lower within 15 s of exposure to 0.5% benzalkonium chloride, within 30 s of exposure to 0.5% alkyldiaminoethyl glycine, or within 1 min of exposure to 0.1% alkyldiaminoethyl glycine, and decreased to about 0.1% with 60 min of exposure to 0.1% benzalkonium chloride or 0.5% chlorhexidine gluconate, but did not decrease to 1% or less with 60 min of exposure to 0.1% chlorhexidine gluconate. There were no effects of 0.1% and 0.2% chlorhexidine gluconate and 0.1% benzalkonium chloride against biofilm cells of B. cepacia, and 0.5% chlorhexidine gluconate, 0.5% benzalkonium chloride and 0.1% alkyldiaminoethyl glycine were barely effective against biofilm cells even after 60-min exposure. On the other hand, both planktonic cells and biofilm cells of B. cepacia were eradicated within 15 s by sodium hypochlorite, povidone-iodine, 80% v/v ethanol, and hot water at 65 degrees C or higher.

  1. Hot nuclear matter

    SciTech Connect

    Chapman, S.

    1992-11-01

    The goal in this thesis is thus twofold: The first is to investigate the feasibility of using heavy ion collisions to create conditions in the laboratory which are ripe for the formation of a quark-gluon plasma. The second is to develop a technique for studying some of the many non-perturbative features of this novel phase of matter.

  2. Effect of medium dependent binding energies on inferring the temperatures and freeze-out density of disassembling hot nuclear matter from cluster yields

    NASA Astrophysics Data System (ADS)

    Shlomo, S.; Röpke, G.; Natowitz, J. B.; Qin, L.; Hagel, K.; Wada, R.; Bonasera, A.

    2009-03-01

    We explore the abundance of light clusters in asymmetric nuclear matter at subsaturation density. With increasing density, binding energies and wave functions are modified due to medium effects. The method of Albergo, Costa, Costanzo, and Rubbino (ACCR) for determining the temperature and free nucleon density of a disassembling hot nuclear source from fragment yields is modified to include, in addition to Coulomb effects and flow, also effects of medium modifications of cluster properties, which become of importance when the nuclear matter density is above 10-3fm-3. We show how the analysis of cluster yields, to infer temperature and nucleon densities, is modified if the shifts in binding energies of in medium clusters are included. Although, at low densities, the temperature calculated from given yields changes only modestly if medium effects are taken into account, larger discrepancies are observed when the nucleon densities are determined from measured yields.

  3. Induced Pluripotent Stem Cells: Emerging Techniques for Nuclear Reprogramming

    PubMed Central

    Han, Ji Woong

    2011-01-01

    Abstract Introduction of four transcription factors, Oct3/4, Sox2, Klf4, and c-Myc, can successfully reprogram somatic cells into embryonic stem (ES)-like cells. These cells, which are referred to as induced pluripotent stem (iPS) cells, closely resemble embryonic stem cells in genomic, cell biologic, and phenotypic characteristics, and the creation of these special cells was a major triumph in cell biology. In contrast to pluripotent stem cells generated by somatic cell nuclear-transfer (SCNT) or ES cells derived from the inner cell mass (ICM) of the blastocyst, direct reprogramming provides a convenient and reliable means of generating pluripotent stem cells. iPS cells have already shown incredible potential for research and for therapeutic applications in regenerative medicine within just a few years of their discovery. In this review, current techniques of generating iPS cells and mechanisms of nuclear reprogramming are reviewed, and the potential for therapeutic applications is discussed. Antioxid. Redox Signal. 15, 1799–1820. PMID:21194386

  4. Animal cloning by somatic cell nuclear transfer.

    PubMed

    Smith, Lawrence C; Yoo, Jae-Gyu

    2009-01-01

    Animal cloning is becoming increasingly useful for its applications in biological inquiry and for its potential use in pharmaceutical, medical, and agricultural fields. Due to the complexity of the numerous steps required in reconstructing oocytes by nuclear transfer, detailed protocols are required to minimize the developmental damages inflicted during these manipulations and to standardize procedures across laboratories. Moreover, because oogenesis and early embryogenesis differ widely among mammalian species, it is essential that protocols be adapted according to each species concerned. Our objective here is to detail the protocols that have been most successful in producing laboratory and domestic animal clones.

  5. Experimental demonstration of hot-carrier photo-current in an InGaAs quantum well solar cell

    SciTech Connect

    Hirst, L. C.; Walters, R. J.; Führer, M. F.; Ekins-Daukes, N. J.

    2014-06-09

    An unambiguous observation of hot-carrier photocurrent from an InGaAs single quantum well solar cell is reported. Simultaneous photo-current and photoluminescence measurements were performed for incident power density 0.04–3 kW cm{sup −2}, lattice temperature 10 K, and forward bias 1.2 V. An order of magnitude photocurrent increase was observed for non-equilibrium hot-carrier temperatures >35 K. This photocurrent activation temperature is consistent with that of equilibrium carriers in a lattice at elevated temperature. The observed hot-carrier photo-current is extracted from the well over an energy selective GaAs barrier, thus integrating two essential components of a hot-carrier solar cell: a hot-carrier absorber and an energy selective contact.

  6. A hot water extract of Curcuma longa inhibits adhesion molecule protein expression and monocyte adhesion to TNF-α-stimulated human endothelial cells.

    PubMed

    Kawasaki, Kengo; Muroyama, Koutarou; Yamamoto, Norio; Murosaki, Shinji

    2015-01-01

    The recruitment of arterial leukocytes to endothelial cells is an important step in the progression of various inflammatory diseases. Therefore, its modulation is thought to be a prospective target for the prevention or treatment of such diseases. Adhesion molecules on endothelial cells are induced by proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), and contribute to the recruitment of leukocytes. In the present study, we investigated the effect of hot water extract of Curcuma longa (WEC) on the protein expression of adhesion molecules, monocyte adhesion induced by TNF-α in human umbilical vascular endothelial cells (HUVECs). Treatment of HUVECs with WEC significantly suppressed both TNF-α-induced protein expression of adhesion molecules and monocyte adhesion. WEC also suppressed phosphorylation and degradation of nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (IκBα) induced by TNF-α in HUVECs, suggesting that WEC inhibits the NF-κB signaling pathway.

  7. Nuclear morphometry and chromatin textural characteristics of basal cell carcinoma.

    PubMed

    Mendaçolli, Paola Jung; Brianezi, Gabrielli; Schmitt, Juliano Vilaverde; Marques, Mariângela Esther Alencar; Miot, Hélio Amante

    2015-01-01

    Histological subtypes of basal cell carcinoma have biological, evolutionary and distinct prognostic behavior. The analysis of characteristics of the nucleus can provide data on their cellular physiology and behavior. The authors of this study evaluated nuclear morphological parameters and textural patterns of chromatin from different subtypes of basal cell carcinoma: nodular (n=37), superficial (n=28) and sclerodermiform (n=28). The parameters were compared between neoplasms' subtypes and with unaffected adjacent basal epithelium. Nuclear area and diameter of sclerodermiform neoplasms were superior to the other subtypes. Chromatin's color intensity and fractal dimension were less intense in superficial subtypes. Nuclear roundness and chromatin's entropy presented lower values in tumors than in normal epithelium. There was significant correlation between morphological and textural variables of normal skin and tumors. Morphometric elements and textural chromatin's homogeneity of basal cell carcinomas may be related to evolutionary, biological and behavior particularities related to each histotype.

  8. Nuclear morphometry and chromatin textural characteristics of basal cell carcinoma*

    PubMed Central

    Mendaçolli, Paola Jung; Brianezi, Gabrielli; Schmitt, Juliano Vilaverde; Marques, Mariângela Esther Alencar; Miot, Hélio Amante

    2015-01-01

    Histological subtypes of basal cell carcinoma have biological, evolutionary and distinct prognostic behavior. The analysis of characteristics of the nucleus can provide data on their cellular physiology and behavior. The authors of this study evaluated nuclear morphological parameters and textural patterns of chromatin from different subtypes of basal cell carcinoma: nodular (n=37), superficial (n=28) and sclerodermiform (n=28). The parameters were compared between neoplasms' subtypes and with unaffected adjacent basal epithelium. Nuclear area and diameter of sclerodermiform neoplasms were superior to the other subtypes. Chromatin's color intensity and fractal dimension were less intense in superficial subtypes. Nuclear roundness and chromatin's entropy presented lower values in tumors than in normal epithelium. There was significant correlation between morphological and textural variables of normal skin and tumors. Morphometric elements and textural chromatin's homogeneity of basal cell carcinomas may be related to evolutionary, biological and behavior particularities related to each histotype. PMID:26734870

  9. Posting in/out technique in KfK hot cells

    SciTech Connect

    Enderlein, H.

    1986-01-01

    The concrete-shielded hot cells of Kernforschungszentrum Karlsruhe GmbH was constructed in the early 1960's. Over the years it became apparent that the original posting in/out technique, described in the paper, was no longer acceptable. Double lit locks of 500-mm diameter were provided in the cell ceiling hatches and unshielded containers fitting these hatches were procured. These actions led to a reduction in personnel exposure. Improvements made to the new posting in/out system since its implementation are discussed.

  10. Nuclear receptor regulation of stemness and stem cell differentiation

    PubMed Central

    Jeong, Yangsik

    2009-01-01

    Stem cells include a diverse number of toti-, pluri-, and multi-potent cells that play important roles in cellular genesis and differentiation, tissue development, and organogenesis. Genetic regulation involving various transcription factors results in the self-renewal and differentiation properties of stem cells. The nuclear receptor (NR) superfamily is composed of 48 ligand-activated transcription factors involved in diverse physiological functions such as metabolism, development, and reproduction. Increasing evidence shows that certain NRs function in regulating stemness or differentiation of embryonic stem (ES) cells and tissue-specific adult stem cells. Here, we review the role of the NR superfamily in various aspects of stem cell biology, including their regulation of stemness, forward- and trans-differentiation events; reprogramming of terminally differentiated cells; and interspecies differences. These studies provide insights into the therapeutic potential of the NR superfamily in stem cell therapy and in treating stem cell-associated diseases (e.g., cancer stem cell). PMID:19696553

  11. Nuclear Motility in Glioma Cells Reveals a Cell-Line Dependent Role of Various Cytoskeletal Components

    PubMed Central

    Kiss, Alexa; Horvath, Peter; Rothballer, Andrea; Kutay, Ulrike; Csucs, Gabor

    2014-01-01

    Nuclear migration is a general term for the movement of the nucleus towards a specific site in the cell. These movements are involved in a number of fundamental biological processes, such as fertilization, cell division, and embryonic development. Despite of its importance, the mechanism of nuclear migration is still poorly understood in mammalian cells. In order to shed light on the mechanical processes underlying nuclear movements, we adapted a micro-patterning based assay. C6 rat and U87 human glioma cells seeded on fibronectin patterns - thereby forced into a bipolar morphology - displayed oscillatory movements of the nucleus or the whole cell, respectively. We found that both the actomyosin system and microtubules are involved in the nuclear/cellular movements of both cell lines, but their contributions are cell-/migration-type specific. Dynein activity was necessary for nuclear migration of C6 cells but active myosin-II was dispensable. On the other hand, coupled nuclear and cellular movements of U87 cells were driven by actomyosin contraction. We explain these cell-line dependent effects by the intrinsic differences in the overall mechanical tension due to the various cytoskeletal elements inside the cell. Our observations showed that the movements of the nucleus and the centrosome are strongly correlated and display large variation, indicating a tight but flexible coupling between them. The data also indicate that the forces responsible for nuclear movements are not acting directly via the centrosome. Based on our observations, we propose a new model for nuclear oscillations in C6 cells in which dynein and microtubule dynamics are the main drivers of nuclear movements. This mechanism is similar to the meiotic nuclear oscillations of Schizosaccharomyces pombe and may be evolutionary conserved. PMID:24691067

  12. Optical Systems To Control Workpiece Position And To Delivery A High Power CO2 Laser Beam Into A Hot Cell

    NASA Astrophysics Data System (ADS)

    Colorito, Filippo; Draghi, S.; Garifo, Luciano; Mantega, F.

    1989-03-01

    The advantages of laser technology for remote machining in hot cell are well known. In this paper are reported the results of an experimental work carried out to determine the attenuation of r radiation by a copper mirror in order to design the optical systems to deliver a high pow er CO2 laser beam into a hot cell. Moreover a system to control the workpiece position with respect to the high power laser beam focal point is described.

  13. Differentiated cells are more efficient than adult stem cells for cloning by somatic cell nuclear transfer.

    PubMed

    Sung, Li-Ying; Gao, Shaorong; Shen, Hongmei; Yu, Hui; Song, Yifang; Smith, Sadie L; Chang, Ching-Chien; Inoue, Kimiko; Kuo, Lynn; Lian, Jin; Li, Ao; Tian, X Cindy; Tuck, David P; Weissman, Sherman M; Yang, Xiangzhong; Cheng, Tao

    2006-11-01

    Since the creation of Dolly via somatic cell nuclear transfer (SCNT), more than a dozen species of mammals have been cloned using this technology. One hypothesis for the limited success of cloning via SCNT (1%-5%) is that the clones are likely to be derived from adult stem cells. Support for this hypothesis comes from the findings that the reproductive cloning efficiency for embryonic stem cells is five to ten times higher than that for somatic cells as donors and that cloned pups cannot be produced directly from cloned embryos derived from differentiated B and T cells or neuronal cells. The question remains as to whether SCNT-derived animal clones can be derived from truly differentiated somatic cells. We tested this hypothesis with mouse hematopoietic cells at different differentiation stages: hematopoietic stem cells, progenitor cells and granulocytes. We found that cloning efficiency increases over the differentiation hierarchy, and terminally differentiated postmitotic granulocytes yield cloned pups with the greatest cloning efficiency.

  14. Mechanism of nuclear movements in a multinucleated cell.

    PubMed

    Gibeaux, Romain; Politi, Antonio Z; Philippsen, Peter; Nédélec, François

    2017-03-01

    Multinucleated cells are important in many organisms, but the mechanisms governing the movements of nuclei sharing a common cytoplasm are not understood. In the hyphae of the plant pathogenic fungus Ashbya gossypii, nuclei move back and forth, occasionally bypassing each other, preventing the formation of nuclear clusters. This is essential for genetic stability. These movements depend on cytoplasmic microtubules emanating from the nuclei that are pulled by dynein motors anchored at the cortex. Using three-dimensional stochastic simulations with parameters constrained by the literature, we predict the cortical anchor density from the characteristics of nuclear movements. The model accounts for the complex nuclear movements seen in vivo, using a minimal set of experimentally determined ingredients. Of interest, these ingredients power the oscillations of the anaphase spindle in budding yeast, but in A. gossypii, this system is not restricted to a specific nuclear cycle stage, possibly as a result of adaptation to hyphal growth and multinuclearity.

  15. Mechanism of nuclear movements in a multinucleated cell

    PubMed Central

    Gibeaux, Romain; Politi, Antonio Z.; Philippsen, Peter; Nédélec, François

    2017-01-01

    Multinucleated cells are important in many organisms, but the mechanisms governing the movements of nuclei sharing a common cytoplasm are not understood. In the hyphae of the plant pathogenic fungus Ashbya gossypii, nuclei move back and forth, occasionally bypassing each other, preventing the formation of nuclear clusters. This is essential for genetic stability. These movements depend on cytoplasmic microtubules emanating from the nuclei that are pulled by dynein motors anchored at the cortex. Using three-dimensional stochastic simulations with parameters constrained by the literature, we predict the cortical anchor density from the characteristics of nuclear movements. The model accounts for the complex nuclear movements seen in vivo, using a minimal set of experimentally determined ingredients. Of interest, these ingredients power the oscillations of the anaphase spindle in budding yeast, but in A. gossypii, this system is not restricted to a specific nuclear cycle stage, possibly as a result of adaptation to hyphal growth and multinuclearity. PMID:28077618

  16. Nuclear responses to depletion of mitochondrial DNA in human cells.

    PubMed

    Li, K; Neufer, P D; Williams, R S

    1995-11-01

    The derivation of human cell lines devoid of mitochondrial (mt) DNA (rho 0) provides an opportunity to study nuclear responses to a chronic impairment of mitochondrial oxidative phosphorylation. Expression of several nuclear genes is induced in human rho 0 cells, including those encoding integral proteins of the mitochondrial inner membrane, intermediate filaments, and ribosomes. In contrast to conditions in which mitochondrial respiration is altered acutely, expression of heat shock proteins and immediate early genes is not induced. Mitochondria from rho 0 cells maintain a transmembrane electrochemical potential and are distributed within the cytoplasm of these cells in a manner indistinguishable from that of wild-type cells. We conclude that a chronic deficiency of mitochondrial oxidative phosphorylation produced by elimination of mtDNA is associated with a different pattern of gene induction than that provoked by other acute or subacute conditions that impair mitochondrial respiration or create energy demands in excess of mitochondrial respiratory capacity.

  17. Nuclear localization of Merkel cell polyomavirus large T antigen in Merkel cell carcinoma

    SciTech Connect

    Nakamura, Tomoyuki; Sato, Yuko; Watanabe, Daisuke; Ito, Hideki; Shimonohara, Nozomi; Tsuji, Takahiro; Nakajima, Noriko; Suzuki, Yoshio; Matsuo, Koma; Nakagawa, Hidemi; Sata, Tetsutaro; Katano, Harutaka

    2010-03-15

    To clarify whether mutations in the large T gene encoded by Merkel cell polyomavirus affect the expression and function of large T antigen in Merkel cell carcinoma cases, we investigated the expression of large T antigen in vitro and in vivo. Immunohistochemistry using a rabbit polyclonal antibody revealed that large T antigen was expressed in the nuclei of Merkel cell carcinoma cells with Merkel cell polyomavirus infection. Deletion mutant analyses identified an Arg-Lys-Arg-Lys sequence (amino acids 277-280) as a nuclear localization signal in large T antigen. Sequence analyses revealed that there were no mutations in the nuclear localization signal in any of the eleven Merkel cell polyomavirus strains examined. Furthermore, stop codons were not observed in the upstream of the nuclear localization signal in any of the Merkel cell carcinoma cases examined. These data suggest that the nuclear localization signal is highly conserved and functional in Merkel cell carcinoma cases.

  18. Nuclear entry of hyperbranched polylysine nanoparticles into cochlear cells

    PubMed Central

    Zhang, Weikai; Zhang, Ya; Löbler, Marian; Schmitz, Klaus-Peter; Ahmad, Aqeel; Pyykkö, Ilmari; Zou, Jing

    2011-01-01

    Background: Gene therapy is a potentially effective therapeutic modality for treating sensorineural hearing loss. Nonviral gene delivery vectors are expected to become extremely safe and convenient, and nanoparticles are the most promising types of vectors. However, infrequent nuclear localization in the cochlear cells limits their application for gene therapy. This study aimed to investigate the potential nuclear entry of hyperbranched polylysine nanoparticles (HPNPs) for gene delivery to cochlear targets. Methods: Rat primary cochlear cells and cochlear explants generated from newborn rats were treated with different concentrations of HPNPs. For the in vivo study, HPNPs were administered to the rats’ round window membranes. Subcellular distribution of HPNPs in different cell populations was observed with confocal microscope 24 hours after administration. Results: Nuclear entry was observed in various cochlear cell types in vitro and in vivo. In the primary cochlear cell culture, concentration-dependent internalization was observed. In the cochlear organotypic culture, abundant HPNPs were found in the modiolus, including the spiral ganglion, organ of Corti, and lateral wall tissues. In the in vivo study, a gradient distribution of HPNPs through different layers of the round window membrane was observed. HPNPs were also distributed in the cells of the middle ear tissue. Additionally, efficient internalization of HPNPs was observed in the organ of Corti and spiral ganglion cells. In primary cochlear cells, HPNPs induced higher transfection efficiency than did Lipofectamine™. Conclusion: These results suggest that HPNPs are potentially an ideal carrier for gene delivery into the cochlea. PMID:21468356

  19. Nuclear targeting peptide scaffolds for lipofection of nondividing mammalian cells.

    PubMed

    Subramanian, A; Ranganathan, P; Diamond, S L

    1999-09-01

    Lipofection of nondividing cells is inefficient because much of the transfected DNA is retained in endosomes, and that which escapes to the cytoplasm enters the nucleus at low rates. To improve the final rate-limiting step of nuclear import, we conjugated a nonclassical nuclear localization signal (NLS) containing the M9 sequence of heterogeneous nuclear ribonucleoprotein (hnRNP) A1, to a cationic peptide scaffold derived from a scrambled sequence of the SV40 T-antigen consensus NLS (ScT). The ScT was added to improve DNA binding of the M9 sequence. Lipofection of confluent endothelium with plasmid complexed with the M9-ScT conjugate resulted in 83% transfection and a 63-fold increase in marker gene expression. The M9-ScT conjugate localized fluorescent plasmid into the nucleus of permeabilized cells, and addition of the nuclear pore blocker wheat germ agglutinin prevented nuclear import. This method of gene transfer may lead to viral- and lipid-free transfection of nondividing cells.

  20. Catastrophic nuclear envelope collapse in cancer cell micronuclei

    PubMed Central

    Hatch, Emily M.; Fischer, Andrew H.; Deerinck, Thomas J.; Hetzer, Martin W.

    2013-01-01

    Summary During mitotic exit missegregated chromosomes can recruit their own nuclear envelope (NE) to form micronuclei (MN). MN have reduced functioning compared to primary nuclei in the same cell, although the two compartments appear to be structurally comparable. Here we show that over 60% of MN undergo an irreversible loss of compartmentalization during interphase due to NE collapse. This disruption of the MN, which is induced by defects in nuclear lamina assembly, drastically reduces nuclear functions and can trigger massive DNA damage. MN disruption is associated with chromatin compaction and invasion of endoplasmic reticulum (ER) tubules into the chromatin. We identified disrupted MN in both major subtypes of human non-small cell lung cancer, suggesting that disrupted MN could be a useful objective biomarker for genomic instability in solid tumors. Our study shows that NE collapse is a key event underlying MN dysfunction and establishes a link between aberrant NE organization and aneuploidy. PMID:23827674

  1. From cloned frogs to patient matched stem cells: induced pluripotency or somatic cell nuclear transfer?

    PubMed

    Yamada, Mitsutoshi; Byrne, James; Egli, Dieter

    2015-10-01

    Nuclear transfer has seen a remarkable comeback in the past few years. Three groups have independently reported the derivation of stem cell lines by somatic cell nuclear transfer, from either adult, neonatal or fetal cells. Though the ability of human oocytes to reprogram somatic cells to stem cells had long been anticipated, success did not arrive on a straightforward path. Little was known about human oocyte biology, and nuclear transfer protocols developed in animals required key changes to become effective with human eggs. By overcoming these challenges, human nuclear transfer research has contributed to a greater understanding of oocyte biology, provided a point of reference for the comparison of induced pluripotent stem cells, and delivered a method for the generation of personalized stem cells with therapeutic potential.

  2. Development of Hot Pressing as a Low Cost Processing Technique for Fuel Cell Fabrication

    SciTech Connect

    Sarin, V

    2003-01-14

    Dependable, plentiful, and economical energy has been the driving force for financial, industrial, and political growth in the US since the mid 19th century. For a country whose progress is so deeply rooted in abundant energy and whose current political agenda involves stabilizing world fossil fuel prices, the development of a reliable, efficient and environmentally friendly power generating source seems compulsory. The maturing of high technology fuel cells may be the panacea the country will find indispensable to free itself from foreign dependence. Fuel cells offer an efficient, combustion-less, virtually pollution-free power source, capable of being sited in downtown urban areas or in remote regions. Fuel cells have few moving parts and run almost silently. Fuel cells are electrochemical devices that convert the chemical energy of a fuel directly to electrical energy. Unlike batteries, which store a finite amount of energy, fuel cells will generate electricity continuously, as long as fuel and oxidant are available to the electrodes. Additionally, fuel cells offer clean, efficient, and reliable power and they can be operated using a variety of fuels. Hence, the fuel cell is an extremely promising technology. Over the course of this research, the fundamental knowledge related to ceramic processing, sintering, and hot pressing to successfully hot press a single operational SOFC in one step has been developed. Ceramic powder processing for each of the components of an SOFC has bene tailored towards this goal. Processing parameter for the electrolyte and cathode have been studied and developed until they converted. Several anode fabrication techniques have been developed. Additionally, a novel anode structured has been developed and refined. These individual processes have been cultivated until a single cell SOFC has been fabricated in one step.

  3. Nuclear distribution of claudin-2 increases cell proliferation in human lung adenocarcinoma cells.

    PubMed

    Ikari, Akira; Watanabe, Ryo; Sato, Tomonari; Taga, Saeko; Shimobaba, Shun; Yamaguchi, Masahiko; Yamazaki, Yasuhiro; Endo, Satoshi; Matsunaga, Toshiyuki; Sugatani, Junko

    2014-09-01

    Claudin-2 is expressed in human lung adenocarcinoma tissue and cell lines, although it is absent in normal lung tissue. However, the role of claudin-2 in cell proliferation and the regulatory mechanism of intracellular distribution remain undefined. Proliferation of human adenocarcinoma A549 cells was decreased by claudin-2 knockdown together with a decrease in the percentage of S phase cells. This knockdown decreased the expression levels of ZONAB and cell cycle regulators. Claudin-2 was distributed in the nucleus in human adenocarcinoma tissues and proliferating A549 cells. The nuclear distribution of ZONAB and percentage of S phase cells were higher in cells exogenously expressing claudin-2 with a nuclear localization signal than in cells expressing claudin-2 with a nuclear export signal. Nuclear claudin-2 formed a complex with ZO-1, ZONAB, and cyclin D1. Nuclear distribution of S208A mutant, a dephosphorylated form of claudin-2, was higher than that of wild type. We suggest that nuclear distribution of claudin-2 is up-regulated by dephosphorylation and claudin-2 serves to retain ZONAB and cyclin D1 in the nucleus, resulting in the enhancement of cell proliferation in lung adenocarcinoma cells.

  4. Nuclear RNA sequencing of the mouse erythroid cell transcriptome.

    PubMed

    Mitchell, Jennifer A; Clay, Ieuan; Umlauf, David; Chen, Chih-Yu; Moir, Catherine A; Eskiw, Christopher H; Schoenfelder, Stefan; Chakalova, Lyubomira; Nagano, Takashi; Fraser, Peter

    2012-01-01

    In addition to protein coding genes a substantial proportion of mammalian genomes are transcribed. However, most transcriptome studies investigate steady-state mRNA levels, ignoring a considerable fraction of the transcribed genome. In addition, steady-state mRNA levels are influenced by both transcriptional and posttranscriptional mechanisms, and thus do not provide a clear picture of transcriptional output. Here, using deep sequencing of nuclear RNAs (nucRNA-Seq) in parallel with chromatin immunoprecipitation sequencing (ChIP-Seq) of active RNA polymerase II, we compared the nuclear transcriptome of mouse anemic spleen erythroid cells with polymerase occupancy on a genome-wide scale. We demonstrate that unspliced transcripts quantified by nucRNA-seq correlate with primary transcript frequencies measured by RNA FISH, but differ from steady-state mRNA levels measured by poly(A)-enriched RNA-seq. Highly expressed protein coding genes showed good correlation between RNAPII occupancy and transcriptional output; however, genome-wide we observed a poor correlation between transcriptional output and RNAPII association. This poor correlation is due to intergenic regions associated with RNAPII which correspond with transcription factor bound regulatory regions and a group of stable, nuclear-retained long non-coding transcripts. In conclusion, sequencing the nuclear transcriptome provides an opportunity to investigate the transcriptional landscape in a given cell type through quantification of unspliced primary transcripts and the identification of nuclear-retained long non-coding RNAs.

  5. Age-dependent deterioration of nuclear pore complexes causes a loss of nuclear integrity in postmitotic cells.

    PubMed

    D'Angelo, Maximiliano A; Raices, Marcela; Panowski, Siler H; Hetzer, Martin W

    2009-01-23

    In dividing cells, nuclear pore complexes (NPCs) disassemble during mitosis and reassemble into the newly forming nuclei. However, the fate of nuclear pores in postmitotic cells is unknown. Here, we show that NPCs, unlike other nuclear structures, do not turn over in differentiated cells. While a subset of NPC components, like Nup153 and Nup50, are continuously exchanged, scaffold nucleoporins, like the Nup107/160 complex, are extremely long-lived and remain incorporated in the nuclear membrane during the entire cellular life span. Besides the lack of nucleoporin expression and NPC turnover, we discovered an age-related deterioration of NPCs, leading to an increase in nuclear permeability and the leaking of cytoplasmic proteins into the nucleus. Our finding that nuclear "leakiness" is dramatically accelerated during aging and that a subset of nucleoporins is oxidatively damaged in old cells suggests that the accumulation of damage at the NPC might be a crucial aging event.

  6. Robust nuclear lamina-based cell classification of aging and senescent cells.

    PubMed

    Righolt, Christiaan H; van 't Hoff, Merel L R; Vermolen, Bart J; Young, Ian T; Raz, Vered

    2011-12-01

    Changes in the shape of the nuclear lamina are exhibited in senescent cells, as well as in cells expressing mutations in lamina genes. To identify cells with defects in the nuclear lamina we developed an imaging method that quantifies the intensity and curvature of the nuclear lamina. We show that this method accurately describes changes in the nuclear lamina. Spatial changes in nuclear lamina coincide with redistribution of lamin A proteins and local reduction in protein mobility in senescent cell. We suggest that local accumulation of lamin A in the nuclear envelope leads to bending of the structure. A quantitative distinction of the nuclear lamina shape in cell populations was found between fresh and senescent cells, and between primary myoblasts from young and old donors. Moreover, with this method mutations in lamina genes were significantly distinct from cells with wild-type genes. We suggest that this method can be applied to identify abnormal cells during aging, in in vitro propagation, and in lamina disorders.

  7. Somatic cell nuclear transfer: pros and cons.

    PubMed

    Sumer, Huseyin; Liu, Jun; Tat, Pollyanna; Heffernan, Corey; Jones, Karen L; Verma, Paul J

    2009-01-01

    Even though the technique of mammalian SCNT is just over a decade old it has already resulted in numerous significant advances. Despite the recent advances in the reprogramming field, SCNT remains the bench-mark for the generation of both genetically unmodified autologous pluripotent stem cells for transplantation and for the production of cloned animals. In this review we will discuss the pros and cons of SCNT, drawing comparisons with other reprogramming methods.

  8. Cell-fusion method to visualize interphase nuclear pore formation.

    PubMed

    Maeshima, Kazuhiro; Funakoshi, Tomoko; Imamoto, Naoko

    2014-01-01

    In eukaryotic cells, the nucleus is a complex and sophisticated organelle that organizes genomic DNA to support essential cellular functions. The nuclear surface contains many nuclear pore complexes (NPCs), channels for macromolecular transport between the cytoplasm and nucleus. It is well known that the number of NPCs almost doubles during interphase in cycling cells. However, the mechanism of NPC formation is poorly understood, presumably because a practical system for analysis does not exist. The most difficult obstacle in the visualization of interphase NPC formation is that NPCs already exist after nuclear envelope formation, and these existing NPCs interfere with the observation of nascent NPCs. To overcome this obstacle, we developed a novel system using the cell-fusion technique (heterokaryon method), previously also used to analyze the shuttling of macromolecules between the cytoplasm and the nucleus, to visualize the newly synthesized interphase NPCs. In addition, we used a photobleaching approach that validated the cell-fusion method. We recently used these methods to demonstrate the role of cyclin-dependent protein kinases and of Pom121 in interphase NPC formation in cycling human cells. Here, we describe the details of the cell-fusion approach and compare the system with other NPC formation visualization methods. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Verification survey of the hot cell facility site, General Atomics, San Diego, California

    SciTech Connect

    Adams, W. C.

    2000-06-30

    From 1958, General Atomics maintain a hot cell facility in support of government-funded research into reactor fuels. As the use of the facility declined, General Atomics entered into an agreement with DOE to dismantle the facility and decontaminate and decommission (D&D) the site so that it could made available for unrestricted use. The Environmental Survey and Site Assessment Program (ESSAP) was requested to verify the final radiological status of the D&D effort. This is the report of ESSAP survey and verification activities conducted at the San Diego site from November 1999 through March 2000.

  10. Particle-in-cell simulations of hot electron generation using defocused laser light in cone targets

    SciTech Connect

    Yang, Lei; Pasley, John

    2016-08-15

    The effects of defocusing a high intensity pulse of laser light on the generation of hot electrons in a cone are investigated using particle-in-cell simulations. The results indicate that defocused laser light can soften the electron energy spectrum and increase the coupling efficiency compared to the use of a laser in tight focus. It is shown that this is a consequence of the density profile of plasma produced by the laser prepulse, which is less dense in the case of the defocused laser. The relevance of this result to fast ignition inertial confinement fusion is discussed.

  11. Bright narrowband biphoton generation from a hot rubidium atomic vapor cell

    NASA Astrophysics Data System (ADS)

    Zhu, Lingbang; Guo, Xianxin; Shu, Chi; Jeong, Heejeong; Du, Shengwang

    2017-04-01

    We demonstrate the generation of high-quality narrowband biphotons from a Doppler-broadened hot rubidium atomic vapor cell. Choosing a double-Λ atomic energy level scheme for optimizing both spontaneous four-wave mixing nonlinear parametric interaction and electromagnetically induced transparency (EIT), we achieve a biphoton spectral brightness as high as 14 000 s-1 MHz-1. Meanwhile, we apply a spatially tailored optical pumping beam for reduction of the Raman noise and obtain a violation of the Cauchy-Schwarz inequality by a factor of 1023.

  12. Nuclear DNA Content Varies with Cell Size across Human Cell Types

    PubMed Central

    Gillooly, James F.; Hein, Andrew; Damiani, Rachel

    2015-01-01

    Variation in the size of cells, and the DNA they contain, is a basic feature of multicellular organisms that affects countless aspects of their structure and function. Within humans, cell size is known to vary by several orders of magnitude, and differences in nuclear DNA content among cells have been frequently observed. Using published data, here we describe how the quantity of nuclear DNA across 19 different human cell types increases with cell volume. This observed increase is similar to intraspecific relationships between DNA content and cell volume in other species, and interspecific relationships between diploid genome size and cell volume. Thus, we speculate that the quantity of nuclear DNA content in somatic cells of humans is perhaps best viewed as a distribution of values that reflects cell size distributions, rather than as a single, immutable quantity. PMID:26134319

  13. Relationship between nuclear and cytoplasmic RNA in Drosophilia cells.

    PubMed

    Levy, B; McCarthy, B J

    1976-06-01

    Polyadenylated RNA was isolated from nuclei of cultured Drosophila cells, Schneider's line 2, and used as a template to synthesize a complementary DNA probe. Hybridization experiments were performed to study the relationship between nuclear and cytoplasmic RNA. About two-thirds of the nuclear polyadenylated RNA sequences exist in the cytoplasm. Experiments with fractionated cDNA probes demonstrated that RNA sequences that are frequent in the nucleus are also abundant in the cytoplasm. These findings are consistent with a precursor-product relationship in which some polyadenylated molecules in the nucleus are destined for the cytoplasm while other sequences are polyadenylated but not transferred.

  14. Characterization of Aes nuclear foci in colorectal cancer cells.

    PubMed

    Itatani, Yoshiro; Sonoshita, Masahiro; Kakizaki, Fumihiko; Okawa, Katsuya; Stifani, Stefano; Itoh, Hideaki; Sakai, Yoshiharu; Taketo, M Mark

    2016-01-01

    Amino-terminal enhancer of split (Aes) is a member of Groucho/Transducin-like enhancer (TLE) family. Aes is a recently found metastasis suppressor of colorectal cancer (CRC) that inhibits Notch signalling, and forms nuclear foci together with TLE1. Although some Notch-associated proteins are known to form subnuclear bodies, little is known regarding the dynamics or functions of these structures. Here, we show that Aes nuclear foci in CRC observed under an electron microscope are in a rather amorphous structure, lacking surrounding membrane. Investigation of their behaviour during the cell cycle by time-lapse cinematography showed that Aes nuclear foci dissolve during mitosis and reassemble after completion of cytokinesis. We have also found that heat shock cognate 70 (HSC70) is an essential component of Aes foci. Pharmacological inhibition of the HSC70 ATPase activity with VER155008 reduces Aes focus formation. These results provide insight into the understanding of Aes-mediated inhibition of Notch signalling.

  15. TERT promoter hot spot mutations are frequent in Indian cervical and oral squamous cell carcinomas.

    PubMed

    Vinothkumar, Vilvanathan; Arunkumar, Ganesan; Revathidevi, Sundaramoorthy; Arun, Kanagaraj; Manikandan, Mayakannan; Rao, Arunagiri Kuha Deva Magendhra; Rajkumar, Kottayasamy Seenivasagam; Ajay, Chandrasekar; Rajaraman, Ramamurthy; Ramani, Rajendren; Murugan, Avaniyapuram Kannan; Munirajan, Arasambattu Kannan

    2016-06-01

    Squamous cell carcinoma (SCC) of the uterine cervix and oral cavity are most common cancers in India. Telomerase reverse transcriptase (TERT) overexpression is one of the hallmarks for cancer, and activation through promoter mutation C228T and C250T has been reported in variety of tumors and often shown to be associated with aggressive tumors. In the present study, we analyzed these two hot spot mutations in 181 primary tumors of the uterine cervix and oral cavity by direct DNA sequencing and correlated with patient's clinicopathological characteristics. We found relatively high frequency of TERT hot spot mutations in both cervical [21.4 % (30/140)] and oral [31.7 % (13/41)] squamous cell carcinomas. In cervical cancer, TERT promoter mutations were more prevalent (25 %) in human papilloma virus (HPV)-negative cases compared to HPV-positive cases (20.6 %), and both TERT promoter mutation and HPV infection were more commonly observed in advanced stage tumors (77 %). Similarly, the poor and moderately differentiated tumors of the uterine cervix had both the TERT hot spot mutations and HPV (16 and 18) at higher frequency (95.7 %). Interestingly, we observed eight homozygous mutations (six 228TT and two 250TT) only in cervical tumors, and all of them were found to be positive for high-risk HPV. To the best of our knowledge, this is the first study from India reporting high prevalence of TERT promoter mutations in primary tumors of the uterine cervix and oral cavity. Our results suggest that TERT reactivation through promoter mutation either alone or in association with the HPV oncogenes (E6 and E7) could play an important role in the carcinogenesis of cervical and oral cancers.

  16. Proliferating cell nuclear antigen/cyclin in cultured human keratinocytes.

    PubMed

    Okada, N; Miyagawa, S; Steinberg, M L; Yoshikawa, K

    1990-09-01

    Expression of proliferating cell nuclear antigen (PCNA)/cyclin in cultured human keratinocytes was studied using an antibody from an SLE patient as the reagent. By indirect immunofluorescence staining, SV40-transformed human keratinocytes expressed PCNA/cyclin in 40-45% of the cells as a nulcear granular fluorescence. After synchronization of these cells, their nuclear distribution pattern during the S phase was sequential and showed a clear correlation with DNA synthesis. Primary cultured keratinocytes grown in high Ca+ medium expressed PCNA/cyclin in 10-15% of the cells with a similar staining pattern. These positively stained cells were confined to the basal and immediate suprabasal layers of the stratified culture sheet. The keratinocytes disaggregated by trypsin were separated according to cell size through a screen of Nitex monofilament cloth. The cells smaller than 15 microns in diameter synthesized abundant PCNA/cyclin, while the larger cells expressed very low levels. These results indicate that the expression of PCNA/cyclin correlates with DNA synthesis in cultured keratinocytes, but is not associated with their differentiation process.

  17. Investigation of the basic physics of high efficiency semiconductor hot carrier solar cell

    NASA Technical Reports Server (NTRS)

    Alfano, R. R.; Wang, W. B.; Mohaidat, J. M.; Cavicchia, M. A.; Raisky, O. Y.

    1995-01-01

    The main purpose of this research program is to investigate potential semiconductor materials and their multi-band-gap MQW (multiple quantum wells) structures for high efficiency solar cells for aerospace and commercial applications. The absorption and PL (photoluminescence) spectra, the carrier dynamics, and band structures have been investigated for semiconductors of InP, GaP, GaInP, and InGaAsP/InP MQW structures, and for semiconductors of GaAs and AlGaAs by previous measurements. The barrier potential design criteria for achieving maximum energy conversion efficiency, and the resonant tunneling time as a function of barrier width in high efficiency MQW solar cell structures have also been investigated in the first two years. Based on previous carrier dynamics measurements and the time-dependent short circuit current density calculations, an InAs/InGaAs - InGaAs/GaAs - GaAs/AlGaAs MQW solar cell structure with 15 bandgaps has been designed. The absorption and PL spectra in InGaAsP/InP bulk and MQW structures were measured at room temperature and 77 K with different pump wavelength and intensity, to search for resonant states that may affect the solar cell activities. Time-resolved IR absorption for InGaAsP/InP bulk and MQW structures has been measured by femtosecond visible-pump and IR-probe absorption spectroscopy. This, with the absorption and PL measurements, will be helpful to understand the basic physics and device performance in multi-bandgap InAs/InGaAs - InGaAs/InP - InP/InGaP MQW solar cells. In particular, the lifetime of the photoexcited hot electrons is an important parameter for the device operation of InGaAsP/InP MQW solar cells working in the resonant tunneling conditions. Lastly, time evolution of the hot electron relaxation in GaAs has been measured in the temperature range of 4 K through 288 K using femtosecond pump-IR-probe absorption technique. The temperature dependence of the hot electron relaxation time in the X valley has been measured.

  18. Investigation of the basic physics of high efficiency semiconductor hot carrier solar cell

    NASA Astrophysics Data System (ADS)

    Alfano, R. R.; Wang, W. B.; Mohaidat, J. M.; Cavicchia, M. A.; Raisky, O. Y.

    1995-05-01

    The main purpose of this research program is to investigate potential semiconductor materials and their multi-band-gap MQW (multiple quantum wells) structures for high efficiency solar cells for aerospace and commercial applications. The absorption and PL (photoluminescence) spectra, the carrier dynamics, and band structures have been investigated for semiconductors of InP, GaP, GaInP, and InGaAsP/InP MQW structures, and for semiconductors of GaAs and AlGaAs by previous measurements. The barrier potential design criteria for achieving maximum energy conversion efficiency, and the resonant tunneling time as a function of barrier width in high efficiency MQW solar cell structures have also been investigated in the first two years. Based on previous carrier dynamics measurements and the time-dependent short circuit current density calculations, an InAs/InGaAs - InGaAs/GaAs - GaAs/AlGaAs MQW solar cell structure with 15 bandgaps has been designed. The absorption and PL spectra in InGaAsP/InP bulk and MQW structures were measured at room temperature and 77 K with different pump wavelength and intensity, to search for resonant states that may affect the solar cell activities. Time-resolved IR absorption for InGaAsP/InP bulk and MQW structures has been measured by femtosecond visible-pump and IR-probe absorption spectroscopy. This, with the absorption and PL measurements, will be helpful to understand the basic physics and device performance in multi-bandgap InAs/InGaAs - InGaAs/InP - InP/InGaP MQW solar cells. In particular, the lifetime of the photoexcited hot electrons is an important parameter for the device operation of InGaAsP/InP MQW solar cells working in the resonant tunneling conditions. Lastly, time evolution of the hot electron relaxation in GaAs has been measured in the temperature range of 4 K through 288 K using femtosecond pump-IR-probe absorption technique. The temperature dependence of the hot electron relaxation time in the X valley has been measured.

  19. A high throughput approach for analysis of cell nuclear deformability at single cell level

    NASA Astrophysics Data System (ADS)

    Ermis, Menekse; Akkaynak, Derya; Chen, Pu; Demirci, Utkan; Hasirci, Vasif

    2016-11-01

    Various physiological and pathological processes, such as cell differentiation, migration, attachment, and metastasis are highly dependent on nuclear elasticity. Nuclear morphology directly reflects the elasticity of the nucleus. We propose that quantification of changes in nuclear morphology on surfaces with defined topography will enable us to assess nuclear elasticity and deformability. Here, we used soft lithography techniques to produce 3 dimensional (3-D) cell culture substrates decorated with micron sized pillar structures of variable aspect ratios and dimensions to induce changes in cellular and nuclear morphology. We developed a high content image analysis algorithm to quantify changes in nuclear morphology at the single-cell level in response to physical cues from the 3-D culture substrate. We present that nuclear stiffness can be used as a physical parameter to evaluate cancer cells based on their lineage and in comparison to non-cancerous cells originating from the same tissue type. This methodology can be exploited for systematic study of mechanical characteristics of large cell populations complementing conventional tools such as atomic force microscopy and nanoindentation.

  20. A high throughput approach for analysis of cell nuclear deformability at single cell level

    PubMed Central

    Ermis, Menekse; Akkaynak, Derya; Chen, Pu; Demirci, Utkan; Hasirci, Vasif

    2016-01-01

    Various physiological and pathological processes, such as cell differentiation, migration, attachment, and metastasis are highly dependent on nuclear elasticity. Nuclear morphology directly reflects the elasticity of the nucleus. We propose that quantification of changes in nuclear morphology on surfaces with defined topography will enable us to assess nuclear elasticity and deformability. Here, we used soft lithography techniques to produce 3 dimensional (3-D) cell culture substrates decorated with micron sized pillar structures of variable aspect ratios and dimensions to induce changes in cellular and nuclear morphology. We developed a high content image analysis algorithm to quantify changes in nuclear morphology at the single-cell level in response to physical cues from the 3-D culture substrate. We present that nuclear stiffness can be used as a physical parameter to evaluate cancer cells based on their lineage and in comparison to non-cancerous cells originating from the same tissue type. This methodology can be exploited for systematic study of mechanical characteristics of large cell populations complementing conventional tools such as atomic force microscopy and nanoindentation. PMID:27841297

  1. Cell for a spent nuclear fuel rack

    SciTech Connect

    Flynn, W.M.

    1987-09-22

    This patent describes a cell for a spent fuel rack, comprising: a sheet metal element having an inside surface and an outside surface, and including a first flat wall portion, a second flat wall portion disposed perpendicularly to the first wall portion, a third flat wall portion disposed perpendicularly to the second wall portion and parallel to the first wall portion, a fourth flat wall portion disposed perpendicularly to the first and third wall portions and parallel to the second wall portion, an elongated bent region joining the first and second wall portions, an additional elongated bent region joining the second and third wall portions, a further elongated bent region joining the third and fourth wall portions, another elongated flat platform portion that is disposed parallel to the fourth wall portion but that is not coplanar with the fourth wall portion, and another elongated flat platform portion that is disposed parallel to the first wall portion but that is not coplanar with the first wall portion; means for joining the another platform portions; four sheets of neutron poison; and four sheet metal wrappers, each securing a respective neutron poison sheet to a respective wall portion.

  2. A combined gas cooled nuclear reactor and fuel cell cycle

    NASA Astrophysics Data System (ADS)

    Palmer, David J.

    Rising oil costs, global warming, national security concerns, economic concerns and escalating energy demands are forcing the engineering communities to explore methods to address these concerns. It is the intention of this thesis to offer a proposal for a novel design of a combined cycle, an advanced nuclear helium reactor/solid oxide fuel cell (SOFC) plant that will help to mitigate some of the above concerns. Moreover, the adoption of this proposal may help to reinvigorate the Nuclear Power industry while providing a practical method to foster the development of a hydrogen economy. Specifically, this thesis concentrates on the importance of the U.S. Nuclear Navy adopting this novel design for its nuclear electric vessels of the future with discussion on efficiency and thermodynamic performance characteristics related to the combined cycle. Thus, the goals and objectives are to develop an innovative combined cycle that provides a solution to the stated concerns and show that it provides superior performance. In order to show performance, it is necessary to develop a rigorous thermodynamic model and computer program to analyze the SOFC in relation with the overall cycle. A large increase in efficiency over the conventional pressurized water reactor cycle is realized. Both sides of the cycle achieve higher efficiencies at partial loads which is extremely important as most naval vessels operate at partial loads as well as the fact that traditional gas turbines operating alone have poor performance at reduced speeds. Furthermore, each side of the cycle provides important benefits to the other side. The high temperature exhaust from the overall exothermic reaction of the fuel cell provides heat for the reheater allowing for an overall increase in power on the nuclear side of the cycle. Likewise, the high temperature helium exiting the nuclear reactor provides a controllable method to stabilize the fuel cell at an optimal temperature band even during transients helping

  3. Subnatural-linewidth biphotons from a Doppler-broadened hot atomic vapour cell

    NASA Astrophysics Data System (ADS)

    Shu, Chi; Chen, Peng; Chow, Tsz Kiu Aaron; Zhu, Lingbang; Xiao, Yanhong; Loy, M. M. T.; Du, Shengwang

    2016-09-01

    Entangled photon pairs, termed as biphotons, have been the benchmark tool for experimental quantum optics. The quantum-network protocols based on photon-atom interfaces have stimulated a great demand for single photons with bandwidth comparable to or narrower than the atomic natural linewidth. In the past decade, laser-cooled atoms have often been used for producing such biphotons, but the apparatus is too large and complicated for engineering. Here we report the generation of subnatural-linewidth (<6 MHz) biphotons from a Doppler-broadened (530 MHz) hot atomic vapour cell. We use on-resonance spontaneous four-wave mixing in a hot paraffin-coated 87Rb vapour cell at 63 °C to produce biphotons with controllable bandwidth (1.9-3.2 MHz) and coherence time (47-94 ns). Our backward phase-matching scheme with spatially separated optical pumping is the key to suppress uncorrelated photons from resonance fluorescence. The result may lead towards miniature narrowband biphoton sources.

  4. Subnatural-linewidth biphotons from a Doppler-broadened hot atomic vapour cell

    PubMed Central

    Shu, Chi; Chen, Peng; Chow, Tsz Kiu Aaron; Zhu, Lingbang; Xiao, Yanhong; Loy, M.M.T.; Du, Shengwang

    2016-01-01

    Entangled photon pairs, termed as biphotons, have been the benchmark tool for experimental quantum optics. The quantum-network protocols based on photon–atom interfaces have stimulated a great demand for single photons with bandwidth comparable to or narrower than the atomic natural linewidth. In the past decade, laser-cooled atoms have often been used for producing such biphotons, but the apparatus is too large and complicated for engineering. Here we report the generation of subnatural-linewidth (<6 MHz) biphotons from a Doppler-broadened (530 MHz) hot atomic vapour cell. We use on-resonance spontaneous four-wave mixing in a hot paraffin-coated 87Rb vapour cell at 63 °C to produce biphotons with controllable bandwidth (1.9–3.2 MHz) and coherence time (47–94 ns). Our backward phase-matching scheme with spatially separated optical pumping is the key to suppress uncorrelated photons from resonance fluorescence. The result may lead towards miniature narrowband biphoton sources. PMID:27658721

  5. Stem cells, nuclear transfer and respect for embryos.

    PubMed

    Clausen, Jens

    2010-01-01

    Harvesting human embryonic stem (hES) cells is a highly controversial field of research because it rests on the destruction of human embryos. Altering the procedure of nuclear transfer (NT) is suggested to generate hES cell lines without ethical obstacles by claiming that no embryo would be involved. While discussing the nature of an embryo and related central questions concerning their moral status and the respect they deserve, this paper argues that the entity created by somatic cell nuclear transfer (SCNT) or altered nuclear transfer (ANT) is an embryo and has the same moral status as a natural embryo. Respect for the embryo is expressed by the ethical principles of proportionality, probability and subsidiarity. This paper argues that the human embryo should only be taken for research with high ranking goals, which are proven in animal experimentation and for which there are no alternatives. This makes ANT obsolete and shows that SCNT to produce hES cells is premature at the present time.

  6. Micronuclei Frequencies and Nuclear Abnormalities in Oral Exfoliated Cells of Nuclear Power Plant Workers

    PubMed Central

    Babannavar, Roopa; Lohra, Abhishek; Kodgi, Ashwin; Bapure, Sunil; Rao, Yogesh; J., Arun; Malghan, Manjunath

    2014-01-01

    Aim: Biomonitoring provides a useful tool to estimate the genetic risk from exposure to genotoxic agents. The aim of this study was to evaluate the frequencies of Micronuclei (MN) and other Nuclear abnormalities (NA) from exfoliated oral mucosal cells in Nuclear Power Station (NPS) workers. Materials and Methods: Micronucleus frequencies in oral exfoliated cells were done from individuals not known to be exposed to either environmental or occupational carcinogens (Group I). Similarly samples were obtained from full-time Nuclear Power Station (NPS) workers with absence of Leukemia and any malignancy (Group II) and workers diagnosed as leukemic patients and undergoing treatment (Group III). Results: There was statistically significant difference between Group I, Group II & Group III. MN and NA frequencies in Leukemic Patients were significantly higher than those in exposed workers &control groups (p < 0.05). Conclusion: MN and other NA reflect genetic changes, events associated with malignancies. Therefore, there is a need to educate those who work in NPS about the potential hazard of occupational exposure and the importance of using protective measures. PMID:25654022

  7. A novel method for somatic cell nuclear transfer to mouse embryonic stem cells.

    PubMed

    Pralong, Danièle; Mrozik, Krzysztof; Occhiodoro, Filomena; Wijesundara, Nishanthi; Sumer, Huseyin; Van Boxtel, Antonius L; Trounson, Alan; Verma, Paul J

    2005-01-01

    Nuclear reprogramming by somatic cell nuclear transfer (SCNT) provides a practical approach for generating autologous pluripotent cells from adult somatic cells. It has been shown that murine somatic cells can also be reprogrammed to a pluripotent-like state by fusion with embryonic stem (ES) cells. Typically, the first step in SCNT involves enucleation of the recipient cell. However, recent evidence suggests that enucleated diploid ES cells may lack reprogramming capabilities. Here we have developed methods whereby larger tetraploid ES cells are first generated by fusion of two mouse ES cell lines transfected with plasmids carrying different antibiotic-resistance cassettes, followed by double antibiotic selection. Tetraploid ES cells grown on tissue culture disks or wells can be efficiently enucleated (up to 99%) using a combination of cytochalasin B treatment and centrifugation, with cytoplasts generated from these cells larger than those obtained from normal diploid ES cells. Also, we show that the enucleation rate is dependent on centrifugation time and cell ploidy. Further, we demonstrate that normal diploid ES cells can be fused to tetraploid ES cells to form heterokaryons, and that selective differential centrifugation conditions can be applied where the tetraploid nucleus is removed while the diploid donor nucleus is retained. This technology opens new avenues for generating autologous, diploid pluripotent cells, and provides a dynamic model for studying nuclear reprogramming in ES cells.

  8. Probing charge transfer and hot carrier dynamics in organic solar cells with terahertz spectroscopy

    NASA Astrophysics Data System (ADS)

    Cunningham, Paul D.; Lane, Paul A.; Melinger, Joseph S.; Esenturk, Okan; Heilweil, Edwin J.

    2016-04-01

    Time-resolved terahertz spectroscopy (TRTS) was used to explore charge generation, transfer, and the role of hot carriers in organic solar cell materials. Two model molecular photovoltaic systems were investigated: with zinc phthalocyanine (ZnPc) or alpha-sexathiophene (α-6T) as the electron donors and buckminsterfullerene (C60) as the electron acceptor. TRTS provides charge carrier conductivity dynamics comprised of changes in both population and mobility. By using time-resolved optical spectroscopy in conjunction with TRTS, these two contributions can be disentangled. The sub-picosecond photo-induced conductivity decay dynamics of C60 were revealed to be caused by auto-ionization: the intrinsic process by which charge is generated in molecular solids. In donor-acceptor blends, the long-lived photo-induced conductivity is used for weight fraction optimization of the constituents. In nanoscale multilayer films, the photo-induced conductivity identifies optimal layer thicknesses. In films of ZnPc/C60, electron transfer from ZnPc yields hot charges that localize and become less mobile as they thermalize. Excitation of high-lying Franck Condon states in C60 followed by hole-transfer to ZnPc similarly produces hot charge carriers that self-localize; charge transfer clearly precedes carrier cooling. This picture is contrasted to charge transfer in α-6T/C60, where hole transfer takes place from a thermalized state and produces equilibrium carriers that do not show characteristic signs of cooling and self-localization. These results illustrate the value of terahertz spectroscopic methods for probing charge transfer reactions.

  9. Probing Charge Transfer and Hot Carrier Dynamics in Organic Solar Cells with Terahertz Spectroscopy.

    PubMed

    Cunningham, Paul D; Lane, Paul A; Melinger, Joseph S; Esenturk, Okan; Heilweil, Edwin J

    2016-01-01

    Time-resolved terahertz spectroscopy (TRTS) was used to explore charge generation, transfer, and the role of hot carriers in organic solar cell materials. Two model molecular photovoltaic systems were investigated: with zinc phthalocyanine (ZnPc) or alpha-sexathiophene (α-6T) as the electron donors and buckminsterfullerene (C60) as the electron acceptor. TRTS provides charge carrier conductivity dynamics comprised of changes in both population and mobility. By using time-resolved optical spectroscopy in conjunction with TRTS, these two contributions can be disentangled. The sub-picosecond photo-induced conductivity decay dynamics of C60 were revealed to be caused by auto-ionization: the intrinsic process by which charge is generated in molecular solids. In donor-acceptor blends, the long-lived photo-induced conductivity is used for weight fraction optimization of the constituents. In nanoscale multilayer films, the photo-induced conductivity identifies optimal layer thicknesses. In films of ZnPc/C60, electron transfer from ZnPc yields hot charges that localize and become less mobile as they thermalize. Excitation of high-lying Franck Condon states in C60 followed by hole-transfer to ZnPc similarly produces hot charge carriers that self-localize; charge transfer clearly precedes carrier cooling. This picture is contrasted to charge transfer in α-6T/C60, where hole transfer takes place from a thermalized state and produces equilibrium carriers that do not show characteristic signs of cooling and self-localization. These results illustrate the value of terahertz spectroscopic methods for probing charge transfer reactions.

  10. Probing Charge Transfer and Hot Carrier Dynamics in Organic Solar Cells with Terahertz Spectroscopy

    PubMed Central

    Cunningham, Paul D.; Lane, Paul A.; Melinger, Joseph S.; Esenturk, Okan; Heilweil, Edwin J.

    2017-01-01

    Time-resolved terahertz spectroscopy (TRTS) was used to explore charge generation, transfer, and the role of hot carriers in organic solar cell materials. Two model molecular photovoltaic systems were investigated: with zinc phthalocyanine (ZnPc) or alpha-sexathiophene (α-6T) as the electron donors and buckminsterfullerene (C60) as the electron acceptor. TRTS provides charge carrier conductivity dynamics comprised of changes in both population and mobility. By using time-resolved optical spectroscopy in conjunction with TRTS, these two contributions can be disentangled. The sub-picosecond photo-induced conductivity decay dynamics of C60 were revealed to be caused by auto-ionization: the intrinsic process by which charge is generated in molecular solids. In donor-acceptor blends, the long-lived photo-induced conductivity is used for weight fraction optimization of the constituents. In nanoscale multilayer films, the photo-induced conductivity identifies optimal layer thicknesses. In films of ZnPc/C60, electron transfer from ZnPc yields hot charges that localize and become less mobile as they thermalize. Excitation of high-lying Franck Condon states in C60 followed by hole-transfer to ZnPc similarly produces hot charge carriers that self-localize; charge transfer clearly precedes carrier cooling. This picture is contrasted to charge transfer in α-6T/C60, where hole transfer takes place from a thermalized state and produces equilibrium carriers that do not show characteristic signs of cooling and self-localization. These results illustrate the value of terahertz spectroscopic methods for probing charge transfer reactions. PMID:28649166

  11. A change in nuclear pore complex composition regulates cell differentiation.

    PubMed

    D'Angelo, Maximiliano A; Gomez-Cavazos, J Sebastian; Mei, Arianna; Lackner, Daniel H; Hetzer, Martin W

    2012-02-14

    Nuclear pore complexes (NPCs) are built from ∼30 different proteins called nucleoporins or Nups. Previous studies have shown that several Nups exhibit cell-type-specific expression and that mutations in NPC components result in tissue-specific diseases. Here we show that a specific change in NPC composition is required for both myogenic and neuronal differentiation. The transmembrane nucleoporin Nup210 is absent in proliferating myoblasts and embryonic stem cells (ESCs) but becomes expressed and incorporated into NPCs during cell differentiation. Preventing Nup210 production by RNAi blocks myogenesis and the differentiation of ESCs into neuroprogenitors. We found that the addition of Nup210 to NPCs does not affect nuclear transport but is required for the induction of genes that are essential for cell differentiation. Our results identify a single change in NPC composition as an essential step in cell differentiation and establish a role for Nup210 in gene expression regulation and cell fate determination. Copyright © 2012 Elsevier Inc. All rights reserved.

  12. Recent advancements in cloning by somatic cell nuclear transfer

    PubMed Central

    Ogura, Atsuo; Inoue, Kimiko; Wakayama, Teruhiko

    2013-01-01

    Somatic cell nuclear transfer (SCNT) cloning is the sole reproductive engineering technology that endows the somatic cell genome with totipotency. Since the first report on the birth of a cloned sheep from adult somatic cells in 1997, many technical improvements in SCNT have been made by using different epigenetic approaches, including enhancement of the levels of histone acetylation in the chromatin of the reconstructed embryos. Although it will take a considerable time before we fully understand the nature of genomic programming and totipotency, we may expect that somatic cell cloning technology will soon become broadly applicable to practical purposes, including medicine, pharmaceutical manufacturing and agriculture. Here we review recent progress in somatic cell cloning, with a special emphasis on epigenetic studies using the laboratory mouse as a model. PMID:23166393

  13. Recent advancements in cloning by somatic cell nuclear transfer.

    PubMed

    Ogura, Atsuo; Inoue, Kimiko; Wakayama, Teruhiko

    2013-01-05

    Somatic cell nuclear transfer (SCNT) cloning is the sole reproductive engineering technology that endows the somatic cell genome with totipotency. Since the first report on the birth of a cloned sheep from adult somatic cells in 1997, many technical improvements in SCNT have been made by using different epigenetic approaches, including enhancement of the levels of histone acetylation in the chromatin of the reconstructed embryos. Although it will take a considerable time before we fully understand the nature of genomic programming and totipotency, we may expect that somatic cell cloning technology will soon become broadly applicable to practical purposes, including medicine, pharmaceutical manufacturing and agriculture. Here we review recent progress in somatic cell cloning, with a special emphasis on epigenetic studies using the laboratory mouse as a model.

  14. Cloning of ES cells and mice by nuclear transfer.

    PubMed

    Wakayama, Sayaka; Kishigami, Satoshi; Wakayama, Teruhiko

    2009-01-01

    We have been able to develop a stable nuclear transfer (NT) method in the mouse, in which donor nuclei are directly injected into the oocyte using a piezo-actuated micromanipulator. Although the piezo unit is a complex tool, once mastered it is of great help not only in NT experiments, but also in almost all other forms of micromanipulation. Using this technique, embryonic stem (ntES) cell lines established from somatic cell nuclei can be generated relatively easily from a variety of mouse genotypes and cell types. Such ntES cells can be used not only for experimental models of human therapeutic cloning but also as a means of preserving mouse genomes instead of preserving germ cells. Here, we describe our most recent protocols for mouse cloning.

  15. Analysis of CRM1-Dependent Nuclear Export in Permeabilized Cells.

    PubMed

    Kehlenbach, Ralph H; Port, Sarah A

    2016-01-01

    Nuclear protein import and export assays in permeabilized cells have been instrumental for the identification of transport factors and for the molecular characterization of nucleocytoplasmic transport pathways. Our original assay to quantitatively analyze CRM1-dependent export was based on stably transfected cells expressing GFP-NFAT. We now present a simplified version of the assay using transiently transfected cells expressing GFP-NFAT or GFP-snurportin1 as a fluorescent export cargo and mCherry-emerin as a marker protein for transfected cells. CRM1- and Ran-dependent export is recapitulated in digitonin-permeabilized cells and quantified by flow cytometry. The assay should be applicable to other combinations of cargo and marker proteins.

  16. Nuclear envelope and genome interactions in cell fate

    PubMed Central

    Talamas, Jessica A.; Capelson, Maya

    2015-01-01

    The eukaryotic cell nucleus houses an organism’s genome and is the location within the cell where all signaling induced and development-driven gene expression programs are ultimately specified. The genome is enclosed and separated from the cytoplasm by the nuclear envelope (NE), a double-lipid membrane bilayer, which contains a large variety of trans-membrane and associated protein complexes. In recent years, research regarding multiple aspects of the cell nucleus points to a highly dynamic and coordinated concert of efforts between chromatin and the NE in regulation of gene expression. Details of how this concert is orchestrated and how it directs cell differentiation and disease are coming to light at a rapid pace. Here we review existing and emerging concepts of how interactions between the genome and the NE may contribute to tissue specific gene expression programs to determine cell fate. PMID:25852741

  17. Murine somatic cell nuclear transfer using reprogrammed donor cells expressing male germ cell-specific genes.

    PubMed

    Kang, Hoin; Park, Jong Im; Roh, Sangho

    2016-01-01

    In vivo-matured mouse oocytes were enucleated, and a single murine embryonic fibroblast (control or reprogrammed by introducing extracts from murine testis tissue, which showed expression of male germ cell-specific genes) was injected into the cytoplasm of the oocytes. The rate of blastocyst development and expression levels of Oct-4, Eomes and Cdx-2 were not significantly different in both experimental groups. However, the expression levels of Nanog, Sox9 and Glut-1 were significantly increased when reprogrammed cells were used as donor nuclei. Increased expression of Nanog can be supportive of complete reprogramming of somatic cell nuclear transfer murine embryos. The present study suggested that donor cells expressing male germ cell-specific genes can be reconstructed and can develop into embryos with normal high expression of developmentally essential genes.

  18. Insights into nuclear dynamics using live-cell imaging approaches.

    PubMed

    Bigley, Rachel B; Payumo, Alexander Y; Alexander, Jeffrey M; Huang, Guo N

    2017-03-01

    The nucleus contains the genetic blueprint of the cell and myriad interactions within this subcellular structure are required for gene regulation. In the current scientific era, characterization of these gene regulatory networks through biochemical techniques coupled with systems-wide 'omic' approaches has become commonplace. However, these strategies are limited because they represent a mere snapshot of the cellular state. To obtain a holistic understanding of nuclear dynamics, relevant molecules must be studied in their native contexts in living systems. Live-cell imaging approaches are capable of providing quantitative assessment of the dynamics of gene regulatory interactions within the nucleus. We survey recent insights into what live-cell imaging approaches have provided the field of nuclear dynamics. In this review, we focus on interactions of DNA with other DNA loci, proteins, RNA, and the nuclear envelope. WIREs Syst Biol Med 2017, 9:e1372. doi: 10.1002/wsbm.1372 For further resources related to this article, please visit the WIREs website.

  19. Nuclear PI3K signaling in cell growth and tumorigenesis

    PubMed Central

    Davis, William J.; Lehmann, Peter Z.; Li, Weimin

    2015-01-01

    The PI3K/Akt signaling pathway is a major driving force in a variety of cellular functions. Dysregulation of this pathway has been implicated in many human diseases including cancer. While the activity of the cytoplasmic PI3K/Akt pathway has been extensively studied, the functions of these molecules and their effector proteins within the nucleus are poorly understood. Harboring key cellular processes such as DNA replication and repair as well as nascent messenger RNA transcription, the nucleus provides a unique compartmental environment for protein–protein and protein–DNA/RNA interactions required for cell survival, growth, and proliferation. Here we summarize recent advances made toward elucidating the nuclear PI3K/Akt signaling cascade and its key components within the nucleus as they pertain to cell growth and tumorigenesis. This review covers the spatial and temporal localization of the major nuclear kinases having PI3K activities and the counteracting phosphatases as well as the role of nuclear PI3K/Akt signaling in mRNA processing and exportation, DNA replication and repair, ribosome biogenesis, cell survival, and tumorigenesis. PMID:25918701

  20. ERK5 and Cell Proliferation: Nuclear Localization Is What Matters.

    PubMed

    Gomez, Nestor; Erazo, Tatiana; Lizcano, Jose M

    2016-01-01

    ERK5, the last MAP kinase family member discovered, is activated by the upstream kinase MEK5 in response to growth factors and stress stimulation. MEK5-ERK5 pathway has been associated to different cellular processes, playing a crucial role in cell proliferation in normal and cancer cells by mechanisms that are both dependent and independent of its kinase activity. Thus, nuclear ERK5 activates transcription factors by either direct phosphorylation or acting as co-activator thanks to a unique transcriptional activation TAD domain located at its C-terminal tail. Consequently, ERK5 has been proposed as an interesting target to tackle different cancers, and either inhibitors of ERK5 activity or silencing the protein have shown antiproliferative activity in cancer cells and to block tumor growth in animal models. Here, we review the different mechanisms involved in ERK5 nuclear translocation and their consequences. Inactive ERK5 resides in the cytosol, forming a complex with Hsp90-Cdc37 superchaperone. In a canonical mechanism, MEK5-dependent activation results in ERK5 C-terminal autophosphorylation, Hsp90 dissociation, and nuclear translocation. This mechanism integrates signals such as growth factors and stresses that activate the MEK5-ERK5 pathway. Importantly, two other mechanisms, MEK5-independent, have been recently described. These mechanisms allow nuclear shuttling of kinase-inactive forms of ERK5. Although lacking kinase activity, these forms activate transcription by interacting with transcription factors through the TAD domain. Both mechanisms also require Hsp90 dissociation previous to nuclear translocation. One mechanism involves phosphorylation of the C-terminal tail of ERK5 by kinases that are activated during mitosis, such as Cyclin-dependent kinase-1. The second mechanism involves overexpression of chaperone Cdc37, an oncogene that is overexpressed in cancers such as prostate adenocarcinoma, where it collaborates with ERK5 to promote cell proliferation

  1. ERK5 and Cell Proliferation: Nuclear Localization Is What Matters

    PubMed Central

    Gomez, Nestor; Erazo, Tatiana; Lizcano, Jose M.

    2016-01-01

    ERK5, the last MAP kinase family member discovered, is activated by the upstream kinase MEK5 in response to growth factors and stress stimulation. MEK5-ERK5 pathway has been associated to different cellular processes, playing a crucial role in cell proliferation in normal and cancer cells by mechanisms that are both dependent and independent of its kinase activity. Thus, nuclear ERK5 activates transcription factors by either direct phosphorylation or acting as co-activator thanks to a unique transcriptional activation TAD domain located at its C-terminal tail. Consequently, ERK5 has been proposed as an interesting target to tackle different cancers, and either inhibitors of ERK5 activity or silencing the protein have shown antiproliferative activity in cancer cells and to block tumor growth in animal models. Here, we review the different mechanisms involved in ERK5 nuclear translocation and their consequences. Inactive ERK5 resides in the cytosol, forming a complex with Hsp90-Cdc37 superchaperone. In a canonical mechanism, MEK5-dependent activation results in ERK5 C-terminal autophosphorylation, Hsp90 dissociation, and nuclear translocation. This mechanism integrates signals such as growth factors and stresses that activate the MEK5-ERK5 pathway. Importantly, two other mechanisms, MEK5-independent, have been recently described. These mechanisms allow nuclear shuttling of kinase-inactive forms of ERK5. Although lacking kinase activity, these forms activate transcription by interacting with transcription factors through the TAD domain. Both mechanisms also require Hsp90 dissociation previous to nuclear translocation. One mechanism involves phosphorylation of the C-terminal tail of ERK5 by kinases that are activated during mitosis, such as Cyclin-dependent kinase-1. The second mechanism involves overexpression of chaperone Cdc37, an oncogene that is overexpressed in cancers such as prostate adenocarcinoma, where it collaborates with ERK5 to promote cell proliferation

  2. Age-dependent deterioration of nuclear pore complexes causes a loss of nuclear integrity in post-mitotic cells

    PubMed Central

    D’Angelo, Maximiliano A.; Raices, Marcela; Panowski, Siler H.; Hetzer, Martin W.

    2009-01-01

    SUMMARY In dividing cells, nuclear pore complexes (NPCs) disassemble during mitosis and reassemble into the newly forming nuclei. However, the fate of these multi-protein transport channels in post-mitotic cells, where the mitotic renewal of pores is absent, is unknown. Here we show that NPCs, unlike other nuclear structures, do not turn over in differentiated cells. While a subset of nuclear pore components, like Nup153 and Nup50, are continuously exchanged at the NPC, scaffold nucleoporins, like the Nup107/160 complex, are extremely long-lived and remain incorporated in the nuclear membrane during the entire lifespan of a cell. In addition to a lack of nucleoporin expression and NPC turnover, we discovered an age-related deterioration of NPCs leading to a loss of the nuclear permeability barrier and the leaking of cytoplasmic proteins into the nuclear compartment. Our finding that nuclear ‘leakiness’ is dramatically accelerated during aging and that a subset of nucleoporins are found to be oxidatively damaged in old cells, suggest that the accumulation of damage at the NPC structure might be a crucial event in age-related loss of nuclear integrity. PMID:19167330

  3. Nuclear lamins and oxidative stress in cell proliferation and longevity.

    PubMed

    Shimi, Takeshi; Goldman, Robert D

    2014-01-01

    In mammalian cells, the nuclear lamina is composed of a complex fibrillar network associated with the inner membrane of the nuclear envelope. The lamina provides mechanical support for the nucleus and functions as the major determinant of its size and shape. At its innermost aspect it associates with peripheral components of chromatin and thereby contributes to the organization of interphase chromosomes. The A- and B-type lamins are the major structural components of the lamina, and numerous mutations in the A-type lamin gene have been shown to cause many types of human diseases collectively known as the laminopathies. These mutations have also been shown to cause a disruption in the normal interactions between the A and B lamin networks. The impact of these mutations on nuclear functions is related to the roles of lamins in regulating various essential processes including DNA synthesis and damage repair, transcription and the regulation of genes involved in the response to oxidative stress. The major cause of oxidative stress is the production of reactive oxygen species (ROS), which is critically important for cell proliferation and longevity. Moderate increases in ROS act to initiate signaling pathways involved in cell proliferation and differentiation, whereas excessive increases in ROS cause oxidative stress, which in turn induces cell death and/or senescence. In this review, we cover current findings about the role of lamins in regulating cell proliferation and longevity through oxidative stress responses and ROS signaling pathways. We also speculate on the involvement of lamins in tumor cell proliferation through the control of ROS metabolism.

  4. Crystal Silicon Heterojunction Solar Cells by Hot-Wire CVD: Preprint

    SciTech Connect

    Wang, Q.; Page, M. R.; Iwaniczko, E.; Xu, Y. Q.; Roybal, L.; Bauer, R.; To, B.; Yuan, H. C.; Duda, A.; Yan, Y. F.

    2008-05-01

    Hot-wire chemical vapor deposition (HWCVD) is a promising technique for fabricating Silicon heterojunction (SHJ) solar cells. In this paper we describe our efforts to increase the open circuit voltage (Voc) while improving the efficiency of these devices. On p-type c-Si float-zone wafers, we used a double heterojunction structure with an amorphous n/i contact to the top surface and an i/p contact to the back surface to obtain an open circuit voltage (Voc) of 679 mV in a 0.9 cm2 cell with an independently confirmed efficiency of 19.1%. This is the best reported performance for a cell of this configuration. We also made progress on p-type CZ wafers and achieved 18.7% independently confirmed efficiency with little degradation under prolong illumination. Our best Voc for a p-type SHJ cell is 0.688 V, which is close to the 691 mV we achieved for SHJ cells on n type c-Si wafers.

  5. Cloning Endangered Felids by Interspecies Somatic Cell Nuclear Transfer.

    PubMed

    Gómez, Martha C; Pope, C Earle

    2015-01-01

    In 2003, the first wild felid was produced by interspecies somatic cell nuclear transfer. Since then other wild felid clone offspring have been produced by using the same technique with minor modifications. This chapter describes detailed protocols used in our laboratory for (1) the isolation, culture, and preparation of fibroblast cells as donor nucleus, and (2) embryo reconstruction with domestic cat enucleated oocytes to produce cloned embryos that develop to the blastocyst stage in vitro and, after transfer into synchronized recipients, establish successful pregnancies.

  6. U.S. Environmental Protection Agency Clean Air Act notice of construction for spent nuclear fuel project - hot conditioning system annex, project W-484

    SciTech Connect

    Baker, S.K., Westinghouse Hanford

    1996-12-10

    This notice of construction (NOC) provides information regarding the source and the estimated quantity of potential airborne radionuclide emissions resulting from the operation of the Hot Conditioning System (HCS) Annex. The construction of the HCS Annex is scheduled to conunence on or about December 1996, and will be completed when the process equipment begins operations. This document serves as a NOC pursuant to the requirements of 40 Code of Federal Regulations (CFR) 61 for the HCS Annex. About 80 percent of the U.S. Department of Energy`s spent nuclear fuel (SNF) inventory is stored under water in the Hanford Site K Basins. Spent nuclear fuel in the K West Basin is contained in closed canisters, while the SNF in the K East Basin is contained in open canisters, which allows release of corrosion products to the K East Basin water. Storage of the current inventory in the K Basins was originally intended to be on an as-needed basis to sustain operation of the N Reactor while the Plutonium-Uranium Extraction (PUREX) Plant was refurbished and restarted. The decision in December 1992 to deactivate the PUREX Plant left approximately 2, 1 00 MT (2,300 tons) of uranium, as part of 1133 N Reactor SNF in the K Basins with no means for near-term removal and processing. The HCS Annex will be constructed as an annex to the Canister Storage Building (CSB) and will contain the hot conditioning equipment. The hot conditioning system (HCS) will release chemically-bound water and will condition (process of using a controlled amount of oxygen to destroy uranium hydride) the exposed uranium surfaces associated with the SNF through oxidation. The HCS Annex will house seven hot conditioning process stations, six operational and one auxiliary, which could be used as a welding area for final closure of the vessel containing the SNF. The auxiliary pit is being evaluated at this time for its usefulness to support other operations that may be needed to ensure proper conditioning of the SNF

  7. Bubbling cell death: A hot air balloon released from the nucleus in the cold

    PubMed Central

    2016-01-01

    Cell death emanating from the nucleus is largely unknown. In our recent study, we determined that when temperature is lowered in the surrounding environment, apoptosis stops and bubbling cell death (BCD) occurs. The study concerns the severity of frostbite. When exposed to severe cold and strong ultraviolet (UV) irradiation, people may suffer serious damages to the skin and internal organs. This ultimately leads to limb amputations, organ failure, and death. BCD is defined as “formation of a single bubble from the nucleus per cell and release of this swelling bubble from the cell surface to extracellular space that causes cell death.” When cells are subjected to UV irradiation and/or brief cold shock (4℃ for 5 min) and then incubated at room temperature or 4℃ for time-lapse microscopy, each cell releases an enlarging nuclear gas bubble containing nitric oxide. Certain cells may simultaneously eject hundreds or thousands of exosome-like particles. Unlike apoptosis, no phosphatidylserine flip-over, mitochondrial apoptosis, damage to Golgi complex, and chromosomal DNA fragmentation are shown in BCD. When the temperature is increased back at 37℃, bubble formation stops and apoptosis restarts. Mechanistically, proapoptotic WW domain-containing oxidoreductase and p53 block the protective TNF receptor adaptor factor 2 that allows nitric oxide synthase 2 to synthesize nitric oxide and bubble formation. In this mini-review, updated knowledge in cell death and the proposed molecular mechanism for BCD are provided. PMID:27075929

  8. Bubbling cell death: A hot air balloon released from the nucleus in the cold.

    PubMed

    Chang, Nan-Shan

    2016-06-01

    Cell death emanating from the nucleus is largely unknown. In our recent study, we determined that when temperature is lowered in the surrounding environment, apoptosis stops and bubbling cell death (BCD) occurs. The study concerns the severity of frostbite. When exposed to severe cold and strong ultraviolet (UV) irradiation, people may suffer serious damages to the skin and internal organs. This ultimately leads to limb amputations, organ failure, and death. BCD is defined as "formation of a single bubble from the nucleus per cell and release of this swelling bubble from the cell surface to extracellular space that causes cell death." When cells are subjected to UV irradiation and/or brief cold shock (4℃ for 5 min) and then incubated at room temperature or 4℃ for time-lapse microscopy, each cell releases an enlarging nuclear gas bubble containing nitric oxide. Certain cells may simultaneously eject hundreds or thousands of exosome-like particles. Unlike apoptosis, no phosphatidylserine flip-over, mitochondrial apoptosis, damage to Golgi complex, and chromosomal DNA fragmentation are shown in BCD. When the temperature is increased back at 37℃, bubble formation stops and apoptosis restarts. Mechanistically, proapoptotic WW domain-containing oxidoreductase and p53 block the protective TNF receptor adaptor factor 2 that allows nitric oxide synthase 2 to synthesize nitric oxide and bubble formation. In this mini-review, updated knowledge in cell death and the proposed molecular mechanism for BCD are provided. © 2016 by the Society for Experimental Biology and Medicine.

  9. Rabbit embryonic stem cell lines derived from fertilized, parthenogenetic or somatic cell nuclear transfer embryos

    SciTech Connect

    Fang, Zhen F.; Gai, Hui; Huang, You Z.; Li, Shan G.; Chen, Xue J.; Shi, Jian J.; Wu, Li; Liu, Ailian; Xu, Ping; Sheng, Hui Z. . E-mail: hzsheng2003@yahoo.com

    2006-11-01

    Embryonic stem cells were isolated from rabbit blastocysts derived from fertilization (conventional rbES cells), parthenogenesis (pES cells) and nuclear transfer (ntES cells), and propagated in a serum-free culture system. Rabbit ES (rbES) cells proliferated for a prolonged time in an undifferentiated state and maintained a normal karyotype. These cells grew in a monolayer with a high nuclear/cytoplasm ratio and contained a high level of alkaline phosphate activity. In addition, rbES cells expressed the pluripotent marker Oct-4, as well as EBAF2, FGF4, TDGF1, but not antigens recognized by antibodies against SSEA-1, SSEA-3, SSEA-4, TRA-1-10 and TRA-1-81. All 3 types of ES cells formed embryoid bodies and generated teratoma that contained tissue types of all three germ layers. rbES cells exhibited a high cloning efficiency, were genetically modified readily and were used as nuclear donors to generate a viable rabbit through somatic cell nuclear transfer. In combination with genetic engineering, the ES cell technology should facilitate the creation of new rabbit lines.

  10. Reshaping the transcriptional frontier: epigenetics and somatic cell nuclear transfer.

    PubMed

    Long, Charles R; Westhusin, Mark E; Golding, Michael C

    2014-02-01

    Somatic-cell nuclear transfer (SCNT) experiments have paved the way to the field of cellular reprogramming. The demonstrated ability to clone over 20 different species to date has proven that the technology is robust but very inefficient, and is prone to developmental anomalies. Yet, the offspring from cloned animals exhibit none of the abnormalities of their parents, suggesting the low efficiency and high developmental mortality are epigenetic in origin. The epigenetic barriers to reprogramming somatic cells into a totipotent embryo capable of developing into a viable offspring are significant and varied. Despite their intimate relationship, chromatin structure and transcription are often not uniformly reprogramed after nuclear transfer, and many cloned embryos develop gene expression profiles that are hybrids between the donor cell and an embryonic blastomere. Recent advances in cellular reprogramming suggest that alteration of donor-cell chromatin structure towards that found in an normal embryo is actually the rate-limiting step in successful development of SCNT embryos. Here we review the literature relevant to the transformation of a somatic-cell nucleus into an embryo capable of full-term development. Interestingly, while resetting somatic transcription and associated epigenetic marks are absolutely required for development of SCNT embryos, life does not demand perfection.

  11. Rhythms of glycerophospholipid synthesis in retinal inner nuclear layer cells.

    PubMed

    Garbarino-Pico, Eduardo; Valdez, Diego J; Contín, María A; Pasquaré, Susana J; Castagnet, Paula I; Giusto, Norma M; Caputto, Beatriz L; Guido, Mario E

    2005-09-01

    The present study demonstrates that the biosynthesis of phospholipids in the inner nuclear layer cells of the chicken retina displays daily rhythms under constant illumination conditions. The vertebrate retina contains circadian oscillators and photoreceptors (PRCs) that temporally regulate its own physiology and synchronize the whole organism to the daily environmental changes. We have previously reported that chicken photoreceptors and retinal ganglion cells (RGCs) present significant daily variations in their phospholipid biosynthesis under constant illumination conditions. Herein, we demonstrate that cell preparations highly enriched in inner nuclear layer cells also exhibit a circadian-regulated phospholipid labeling after the in vivo administration of [(32)P]phosphate or [(3)H]glycerol both in animals maintained under constant darkness or light for at least 48h. In constant darkness, there was a significant incorporation of both precursors into phospholipids with the highest levels of labeling around midday and dusk. In constant light, the labeling of (32)P-phospholipids was also significantly higher during the day and early night whereas the incorporation of [(3)H]glycerol into phospholipids, that indicates de novo biosynthesis, was greater during the day but probably reflecting a higher precursor availability at those phases. We also measured the in vitro activity of phosphatidate phosphohydrolase and diacylglycerol lipase in preparations obtained from the dark condition. The two enzymes exhibited the highest activity levels late in the day. When we assessed the in vitro incorporation of [(14)C]oleate into different lysophospholipids from samples collected at different phases in constant darkness, reaction catalyzed by lysophospholipid acyltransferases II, labeling showed a complex pattern of daily activity. Taken together, these results demonstrate that the biosynthesis of phospholipids in cells of the chicken retinal inner nuclear layer exhibits a daily

  12. Nuclear microprobe imaging of gallium nitrate in cancer cells

    NASA Astrophysics Data System (ADS)

    Ortega, Richard; Suda, Asami; Devès, Guillaume

    2003-09-01

    Gallium nitrate is used in clinical oncology as treatment for hypercalcemia and for cancer that has spread to the bone. Its mechanism of antitumor action has not been fully elucidated yet. The knowledge of the intracellular distribution of anticancer drugs is of particular interest in oncology to better understand their cellular pharmacology. In addition, most metal-based anticancer compounds interact with endogenous trace elements in cells, altering their metabolism. The purpose of this experiment was to examine, by use of nuclear microprobe analysis, the cellular distribution of gallium and endogenous trace elements within cancer cells exposed to gallium nitrate. In a majority of cellular analyses, gallium was found homogeneously distributed in cells following the distribution of carbon. In a smaller number of cells, however, gallium appeared concentrated together with P, Ca and Fe within round structures of about 2-5 μm diameter located in the perinuclear region. These intracellular structures are typical of lysosomial material.

  13. Generation of cloned mice and nuclear transfer embryonic stem cell lines from urine-derived cells.

    PubMed

    Mizutani, Eiji; Torikai, Kohei; Wakayama, Sayaka; Nagatomo, Hiroaki; Ohinata, Yasuhide; Kishigami, Satoshi; Wakayama, Teruhiko

    2016-04-01

    Cloning animals by nuclear transfer provides the opportunity to preserve endangered mammalian species. However, there are risks associated with the collection of donor cells from the body such as accidental injury to or death of the animal. Here, we report the production of cloned mice from urine-derived cells collected noninvasively. Most of the urine-derived cells survived and were available as donors for nuclear transfer without any pretreatment. After nuclear transfer, 38-77% of the reconstructed embryos developed to the morula/blastocyst, in which the cell numbers in the inner cell mass and trophectoderm were similar to those of controls. Male and female cloned mice were delivered from cloned embryos transferred to recipient females, and these cloned animals grew to adulthood and delivered pups naturally when mated with each other. The results suggest that these cloned mice had normal fertility. In additional experiments, 26 nuclear transfer embryonic stem cell lines were established from 108 cloned blastocysts derived from four mouse strains including inbreds and F1 hybrids with relatively high success rates. Thus, cells derived from urine, which can be collected noninvasively, may be used in the rescue of endangered mammalian species by using nuclear transfer without causing injury to the animal.

  14. Generation of cloned mice and nuclear transfer embryonic stem cell lines from urine-derived cells

    PubMed Central

    Mizutani, Eiji; Torikai, Kohei; Wakayama, Sayaka; Nagatomo, Hiroaki; Ohinata, Yasuhide; Kishigami, Satoshi; Wakayama, Teruhiko

    2016-01-01

    Cloning animals by nuclear transfer provides the opportunity to preserve endangered mammalian species. However, there are risks associated with the collection of donor cells from the body such as accidental injury to or death of the animal. Here, we report the production of cloned mice from urine-derived cells collected noninvasively. Most of the urine-derived cells survived and were available as donors for nuclear transfer without any pretreatment. After nuclear transfer, 38–77% of the reconstructed embryos developed to the morula/blastocyst, in which the cell numbers in the inner cell mass and trophectoderm were similar to those of controls. Male and female cloned mice were delivered from cloned embryos transferred to recipient females, and these cloned animals grew to adulthood and delivered pups naturally when mated with each other. The results suggest that these cloned mice had normal fertility. In additional experiments, 26 nuclear transfer embryonic stem cell lines were established from 108 cloned blastocysts derived from four mouse strains including inbreds and F1 hybrids with relatively high success rates. Thus, cells derived from urine, which can be collected noninvasively, may be used in the rescue of endangered mammalian species by using nuclear transfer without causing injury to the animal. PMID:27033801

  15. Experimental evidence of hot carriers solar cell operation in multi-quantum wells heterostructures

    SciTech Connect

    Rodière, Jean; Lombez, Laurent; Le Corre, Alain; Durand, Olivier; Guillemoles, Jean-François

    2015-05-04

    We investigated a semiconductor heterostructure based on InGaAsP multi quantum wells (QWs) using optical characterizations and demonstrate its potential to work as a hot carrier cell absorber. By analyzing photoluminescence spectra, the quasi Fermi level splitting Δμ and the carrier temperature are quantitatively measured as a function of the excitation power. Moreover, both thermodynamics values are measured at the QWs and the barrier emission energy. High values of Δμ are found for both transition, and high carrier temperature values in the QWs. Remarkably, the quasi Fermi level splitting measured at the barrier energy exceeds the absorption threshold of the QWs. This indicates a working condition beyond the classical Shockley-Queisser limit.

  16. Electrical characteristics and hot carrier effects in quantum well solar cells

    NASA Astrophysics Data System (ADS)

    Nguyen, Dac-Trung; Lombez, Laurent; Gibelli, Francois; Paire, Myriam; Boyer-Richard, Soline; Durand, Olivier; Guillemoles, Jean-François

    2017-02-01

    We report on the opto-electrical characterization of quantum-well solar cells designed for generation of hot carriers. Short-circuit current is proportional to laser power in the entire range. Population density, temperature and quasi-Fermi level splitting of photo-generated confined carriers are investigated by fitting the full luminescence spectra using generalized Planck's law. The energy-dependent absorptivity is identified to obtain good fit accuracy and takes into account the absorption of excitons and free carriers in the quantum well. Furthermore, electrical injection and extraction across the barriers modify the temperature of the quantum well carrier population linearly, hinting at the role of barriers as semi-selective high-energy contact.

  17. Hot-carrier solar cells using low-dimensional quantum structures

    NASA Astrophysics Data System (ADS)

    Watanabe, Daiki; Kasamatsu, Naofumi; Harada, Yukihiro; Kita, Takashi

    2014-10-01

    We propose a high-conversion-efficiency solar cell (SC) utilizing the hot carrier (HC) population in an intermediate-band (IB) of a quantum dot superlattice (QDSL) structure. The bandgap of the host semiconductor in this device plays an important role as an energy-selective barrier for HCs in the QDSLs. According to theoretical calculation using the detailed balance model with an air mass 1.5 spectrum, the optimum IB energy is determined by a trade-off relation between the number of HCs with energy exceeding the conduction-band edge and the number of photons absorbed by the valence band-IB transition. Utilizing experimental data of HC temperature in InAs/GaAs QDSLs, the maximum conversion efficiency under maximum concentration (45 900 suns) has been demonstrated to increase by 12.6% as compared with that for a single-junction GaAs SC.

  18. Hot Cell Facility modifications at Sandia National Laboratories to support {sup 99}Mo production

    SciTech Connect

    Vernon, M.; Philbin, J.; Berry, D.

    1997-08-01

    In September, 1996, following the completion of an extensive Environmental Impact Statement (EIS), a record of decision (ROD) was issued by DOE selecting Sandia as the facility to take on the {sup 99}Mo production mission. {sup 99}Mo is the precursor to {sup 99m}Tc which is used in 36,000 medical procedures per day in the US. to meet US {sup 99}Mo medical demands, 20 kCi of {sup 99}Mo must be delivered to the pharmaceutical companies each week. This could be accomplished by the processing of twenty-five targets (total fission product of 15 kCi/target) each week within the SNL Hot Cell Facility (HCF). To accomplish this new mission, significant modifications to the HCF will have to be undertaken. This paper presents a brief history of the HCF, and describes modifications necessary to achieve DOE directives.

  19. Hot-carrier solar cells using low-dimensional quantum structures

    SciTech Connect

    Watanabe, Daiki; Kasamatsu, Naofumi; Harada, Yukihiro; Kita, Takashi

    2014-10-27

    We propose a high-conversion-efficiency solar cell (SC) utilizing the hot carrier (HC) population in an intermediate-band (IB) of a quantum dot superlattice (QDSL) structure. The bandgap of the host semiconductor in this device plays an important role as an energy-selective barrier for HCs in the QDSLs. According to theoretical calculation using the detailed balance model with an air mass 1.5 spectrum, the optimum IB energy is determined by a trade-off relation between the number of HCs with energy exceeding the conduction-band edge and the number of photons absorbed by the valence band−IB transition. Utilizing experimental data of HC temperature in InAs/GaAs QDSLs, the maximum conversion efficiency under maximum concentration (45 900 suns) has been demonstrated to increase by 12.6% as compared with that for a single-junction GaAs SC.

  20. Frequent somatic transfer of mitochondrial DNA into the nuclear genome of human cancer cells.

    PubMed

    Ju, Young Seok; Tubio, Jose M C; Mifsud, William; Fu, Beiyuan; Davies, Helen R; Ramakrishna, Manasa; Li, Yilong; Yates, Lucy; Gundem, Gunes; Tarpey, Patrick S; Behjati, Sam; Papaemmanuil, Elli; Martin, Sancha; Fullam, Anthony; Gerstung, Moritz; Nangalia, Jyoti; Green, Anthony R; Caldas, Carlos; Borg, Åke; Tutt, Andrew; Lee, Ming Ta Michael; van't Veer, Laura J; Tan, Benita K T; Aparicio, Samuel; Span, Paul N; Martens, John W M; Knappskog, Stian; Vincent-Salomon, Anne; Børresen-Dale, Anne-Lise; Eyfjörd, Jórunn Erla; Myklebost, Ola; Flanagan, Adrienne M; Foster, Christopher; Neal, David E; Cooper, Colin; Eeles, Rosalind; Bova, Steven G; Lakhani, Sunil R; Desmedt, Christine; Thomas, Gilles; Richardson, Andrea L; Purdie, Colin A; Thompson, Alastair M; McDermott, Ultan; Yang, Fengtang; Nik-Zainal, Serena; Campbell, Peter J; Stratton, Michael R

    2015-06-01

    Mitochondrial genomes are separated from the nuclear genome for most of the cell cycle by the nuclear double membrane, intervening cytoplasm, and the mitochondrial double membrane. Despite these physical barriers, we show that somatically acquired mitochondrial-nuclear genome fusion sequences are present in cancer cells. Most occur in conjunction with intranuclear genomic rearrangements, and the features of the fusion fragments indicate that nonhomologous end joining and/or replication-dependent DNA double-strand break repair are the dominant mechanisms involved. Remarkably, mitochondrial-nuclear genome fusions occur at a similar rate per base pair of DNA as interchromosomal nuclear rearrangements, indicating the presence of a high frequency of contact between mitochondrial and nuclear DNA in some somatic cells. Transmission of mitochondrial DNA to the nuclear genome occurs in neoplastically transformed cells, but we do not exclude the possibility that some mitochondrial-nuclear DNA fusions observed in cancer occurred years earlier in normal somatic cells.

  1. Frequent somatic transfer of mitochondrial DNA into the nuclear genome of human cancer cells

    PubMed Central

    Ju, Young Seok; Tubio, Jose M.C.; Mifsud, William; Fu, Beiyuan; Davies, Helen R.; Ramakrishna, Manasa; Li, Yilong; Yates, Lucy; Gundem, Gunes; Tarpey, Patrick S.; Behjati, Sam; Papaemmanuil, Elli; Martin, Sancha; Fullam, Anthony; Gerstung, Moritz; Nangalia, Jyoti; Green, Anthony R.; Caldas, Carlos; Borg, Åke; Tutt, Andrew; Lee, Ming Ta Michael; van't Veer, Laura J.; Tan, Benita K.T.; Aparicio, Samuel; Span, Paul N.; Martens, John W.M.; Knappskog, Stian; Vincent-Salomon, Anne; Børresen-Dale, Anne-Lise; Eyfjörd, Jórunn Erla; Flanagan, Adrienne M.; Foster, Christopher; Neal, David E.; Cooper, Colin; Eeles, Rosalind; Lakhani, Sunil R.; Desmedt, Christine; Thomas, Gilles; Richardson, Andrea L.; Purdie, Colin A.; Thompson, Alastair M.; McDermott, Ultan; Yang, Fengtang; Nik-Zainal, Serena; Campbell, Peter J.; Stratton, Michael R.

    2015-01-01

    Mitochondrial genomes are separated from the nuclear genome for most of the cell cycle by the nuclear double membrane, intervening cytoplasm, and the mitochondrial double membrane. Despite these physical barriers, we show that somatically acquired mitochondrial-nuclear genome fusion sequences are present in cancer cells. Most occur in conjunction with intranuclear genomic rearrangements, and the features of the fusion fragments indicate that nonhomologous end joining and/or replication-dependent DNA double-strand break repair are the dominant mechanisms involved. Remarkably, mitochondrial-nuclear genome fusions occur at a similar rate per base pair of DNA as interchromosomal nuclear rearrangements, indicating the presence of a high frequency of contact between mitochondrial and nuclear DNA in some somatic cells. Transmission of mitochondrial DNA to the nuclear genome occurs in neoplastically transformed cells, but we do not exclude the possibility that some mitochondrial-nuclear DNA fusions observed in cancer occurred years earlier in normal somatic cells. PMID:25963125

  2. HOT CELL SYSTEM FOR DETERMINING FISSION GAS RETENTION IN METALLIC FUELS

    SciTech Connect

    Sell, D. A.; Baily, C. E.; Malewitz, T. J.; Medvedev, P. G.; Porter, D. L.; Hilton, B. A.

    2016-09-01

    A system has been developed to perform measurements on irradiated, sodium bonded-metallic fuel elements to determine the amount of fission gas retained in the fuel material after release of the gas to the element plenum. During irradiation of metallic fuel elements, most of the fission gas developed is released from the fuel and captured in the gas plenums of the fuel elements. A significant amount of fission gas, however, remains captured in closed porosities which develop in the fuel during irradiation. Additionally, some gas is trapped in open porosity but sealed off from the plenum by frozen bond sodium after the element has cooled in the hot cell. The Retained fission Gas (RFG) system has been designed, tested and implemented to capture and measure the quantity of retained fission gas in characterized cut pieces of sodium bonded metallic fuel. Fuel pieces are loaded into the apparatus along with a prescribed amount of iron powder, which is used to create a relatively low melting, eutectic composition as the iron diffuses into the fuel. The apparatus is sealed, evacuated, and then heated to temperatures in excess of the eutectic melting point. Retained fission gas release is monitored by pressure transducers during the heating phase, thus monitoring for release of fission gas as first the bond sodium melts and then the fuel. A separate hot cell system is used to sample the gas in the apparatus and also characterize the volume of the apparatus thus permitting the calculation of the total fission gas release from the fuel element samples along with analysis of the gas composition.

  3. Multiphysics Thermal-Fluid Analysis of a Non-Nuclear Tester for Hot-Hydrogen Materials Development

    NASA Technical Reports Server (NTRS)

    Wang, Ten-See; Foote, John; Litchford, Ron

    2006-01-01

    The objective of this effort is to analyze the thermal field of a non-nuclear tester, as a first step towards developing efficient and accurate multiphysics, thermo-fluid computational methodology to predict environments for hypothetical solid-core, nuclear thermal engine thrust chamber design and analysis. The computational methodology is based on a multidimensional, finite-volume, turbulent, chemically reacting, radiating, unstructured-grid, and pressure-based formulation. The multiphysics invoked in this study include hydrogen dissociation kinetics and thermodynamics, turbulent flow, convective, radiative and conjugate heat transfers.

  4. Variational Theory of Hot Dense Matter

    ERIC Educational Resources Information Center

    Mukherjee, Abhishek

    2009-01-01

    We develop a variational theory of hot nuclear matter in neutron stars and supernovae. It can also be used to study charged, hot nuclear matter which may be produced in heavy-ion collisions. This theory is a generalization of the variational theory of cold nuclear and neutron star matter based on realistic models of nuclear forces and pair…

  5. Variational Theory of Hot Dense Matter

    ERIC Educational Resources Information Center

    Mukherjee, Abhishek

    2009-01-01

    We develop a variational theory of hot nuclear matter in neutron stars and supernovae. It can also be used to study charged, hot nuclear matter which may be produced in heavy-ion collisions. This theory is a generalization of the variational theory of cold nuclear and neutron star matter based on realistic models of nuclear forces and pair…

  6. Cell Mechanosensitivity is Enabled by the LINC Nuclear Complex

    PubMed Central

    Uzer, Gunes; Rubin, Clinton T.; Rubin, Janet

    2016-01-01

    Mechanoresponses in mesenchymal stem cells (MSCs) guide both differentiation and function. In this review, we focus on advances in0 our understanding of how the cytoplasmic cytoskeleton, nuclear envelope and nucleoskeleton, which are connected via LINC (Linker of Nucleoskeleton and Cytoskeleton) complexes, are emerging as an integrated dynamic signaling platform to regulate MSC mechanobiology. This dynamic interconnectivity affects mechanical signaling and transfer of signals into the nucleus. In this way, nuclear and LINC-mediated cytoskeletal connectivity play a critical role in maintaining mechanical signaling that affects MSC fate by serving as both mechanosensory and mechanoresponsive structures. We review disease and age related compromises of LINC complexes and nucleoskeleton that contribute to the etiology of musculoskeletal diseases. Finally we invite the idea that acquired dysfunctions of LINC might be a contributing factor to conditions such as aging, microgravity and osteoporosis and discuss potential mechanical strategies to modulate LINC connectivity to combat these conditions. PMID:27326387

  7. The nuclear envelope in the crystalline lens fiber cell.

    PubMed

    Harding, C V; Susan, S R

    1976-05-01

    Rabbit lenses which have been fixed, dehydrated, and dried by a critical-point drying method, can be fractured through the cytoplasm of the differentiating lens fibers, exposing the cell nuclei. The fracture, under these conditions, causes a complete separation of the two membranes of the nuclear envelope from one another, thus exposing entire membrane surfaces (those which line the perinuclear space). These surfaces are not seen in their entirety in typical freeze-fracture or freeze-etch preparations, and consequently have not been described previously. The exposed membrane surfaces which line the perinuclear space have numerous convex structures of approximately 1,000 A, and some larger more irregularly shaped structures. These appear to be fragments of the nuclear pore complexes. Differences in these structures between young fibers and those nearing completion of differentiation is suggested.

  8. Effect of hot-pressing on an electrospun TiO2 electrode for dye-sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Kim, Jeong-Hwa; Sung, Shi-Joon; Kim, Kang-Pil; Hwang, Dae-Kue

    2014-07-01

    A hot-pressing process is shown to enhance the adhesion of TiO2 nanofibers electrospun onto fluorine-doped tin oxide substrates for use in dye-sensitized solar cells. By using the resulting electrochemical impedance spectroscopic characteristics, the application of 7, 14, and 21 MPa of pressure was found to result in calculated lifetimes of 7.48, 15.13, and 13.74 ms, respectively. From the results of a thermogravimetric analysis, a stepped heat treatment was developed for calcination of the hot-pressed TiO2 electrodes. Hot-pressing at 14 MPa and 100 °C yielded the dye-sensitized solar cell with the highest energy conversion efficiency.

  9. BLACK HOLE-NEUTRON STAR MERGERS WITH A HOT NUCLEAR EQUATION OF STATE: OUTFLOW AND NEUTRINO-COOLED DISK FOR A LOW-MASS, HIGH-SPIN CASE

    SciTech Connect

    Deaton, M. Brett; Duez, Matthew D.; Foucart, Francois; O'Connor, Evan; Ott, Christian D.; Scheel, Mark A.; Szilagyi, Bela; Kidder, Lawrence E.; Muhlberger, Curran D. E-mail: m.duez@wsu.edu

    2013-10-10

    Neutrino emission significantly affects the evolution of the accretion tori formed in black hole-neutron star mergers. It removes energy from the disk, alters its composition, and provides a potential power source for a gamma-ray burst. To study these effects, simulations in general relativity with a hot microphysical equation of state (EOS) and neutrino feedback are needed. We present the first such simulation, using a neutrino leakage scheme for cooling to capture the most essential effects and considering a moderate mass (1.4 M{sub ☉} neutron star, 5.6 M{sub ☉} black hole), high-spin (black hole J/M {sup 2} = 0.9) system with the K{sub 0} = 220 MeV Lattimer-Swesty EOS. We find that about 0.08 M{sub ☉} of nuclear matter is ejected from the system, while another 0.3 M{sub ☉} forms a hot, compact accretion disk. The primary effects of the escaping neutrinos are (1) to make the disk much denser and more compact, (2) to cause the average electron fraction Y{sub e} of the disk to rise to about 0.2 and then gradually decrease again, and (3) to gradually cool the disk. The disk is initially hot (T ∼ 6 MeV) and luminous in neutrinos (L{sub ν} ∼ 10{sup 54} erg s{sup –1}), but the neutrino luminosity decreases by an order of magnitude over 50 ms of post-merger evolution.

  10. Nuclear morphometry in canine acanthomatous ameloblastomas and squamous cell carcinomas.

    PubMed

    Martano, M; Damiano, S; Restucci, B; Paciello, O; Russo, V; Maiolino, P

    2006-01-01

    The aim of this study was to evaluate whether morphometrical analysis can be of diagnostic value for canine acanthomatous ameloblastoma. We calculated, by means of an automated image analyser, some morphometric nuclear parameters, in particular: mean nuclear area (MNA), mean nuclear perimeter (MNP), maximum and minimum diameters (MDx and MDm) coefficient of variation of the nuclear area (NACV), largest to smallest dimension ratio (LS ratio), and form factor (FF), in 8 canine acanthomatous ameloblastomas, and we compared these morphometric data to those of 13 squamous cell carcinomas of canine gingiva. The results indicated a progressive increase of the MNA, NACV, MNP and MDm proceeding from acanthomatous ameloblastomas (MNA: 42.11+/-8.74; NACV: 28,36+/-7,23; MNP: 24.18+/- 2.68; MDm: 5.69+/-0.49) to squamous cell carcinomas (MNA:49,69+/-9,10; NACV: 30,89+/-7,75; MNP: 25.63+/-2.54; MDm: 6.64+/-0.73). On the contrary, the LS ratio and the FF resulted greater in acanthomatous ameloblastomas (LS ratio: 1,63+/-0,12; FF: 1,13+/-0,002) than in SCCs (LS ratio: 1,40+/-0,12; FF:0.91+/-0.38). Moreover, the MNA, MNP,MDx and MDm resulted similar (MNA: p=0.89; MNP: p=0,65; MDm: p=0,16; MDx: p=0,13) in a subset of four acanthomatous ameloblastomas with cellular atypia (MNA:49,01+/-6,88; MNP: 26,28+/-1,99; MDm: 6.08+/-0.41; MDx: 10.18+/-0.88) and in squamous cell carcinomas (MNA:49.69+/-9,10; MNP: 25.63+/-2.54; MDm: 6.64+/-0.73; MDx: 9.26+/-1.05). While the NACV values resulted higher in typical acanthomatous ameloblastoma (29,99+/-6,06) than in atypical acanthomatous ameloblastoma (26,74+/-8,84) and similar to those of the SCCs (30,89+/-7,75). These results seem to confirm that acanthomatous ameloblastoma is a malignant or potentially malignant lesion and emphasizes that nuclear morphometry analysis can be an useful diagnostic and prognostic method in canine oral pathology.

  11. Rate of energy change of proton traversing in hot high-Z plasmas due to nuclear collision

    NASA Astrophysics Data System (ADS)

    He, Bin; Wang, Jian-Guo

    2015-12-01

    The rate of change of the energy of the projectile proton moving in hot Au plasmas due to the elastic binary collision between the projectile and the target ion at different density and temperature is studied based on the potential from ionic sphere model. It is found that the proton may obtain energy when its kinetic energy is less than the plasma temperature, which means that the proton and the target ion are in thermal equilibrium when the kinetic energy of the proton is around the plasma temperature. The well known model (Phys. Rev. 126, 1 (1962)) is found not to work in hot high-Z plasmas. The reason for this is explored and found relevant to the very small thermal velocity of the high-Z ion compared with the projectile. This leads to the failure to ignore the dependence of the Coulomb logarithm upon the relative velocity between the projectile and the target ion. A revised model is proposed by us and found to work well while the revised model (Phys. Rev. A 29, 2145 (1984)) is unsatisfactory in this case.

  12. Nuclear removal during terminal lens fiber cell differentiation requires CDK1 activity: appropriating mitosis-related nuclear disassembly

    PubMed Central

    Chaffee, Blake R.; Shang, Fu; Chang, Min-Lee; Clement, Tracy M.; Eddy, Edward M.; Wagner, Brad D.; Nakahara, Masaki; Nagata, Shigekazu; Robinson, Michael L.; Taylor, Allen

    2014-01-01

    Lens epithelial cells and early lens fiber cells contain the typical complement of intracellular organelles. However, as lens fiber cells mature they must destroy their organelles, including nuclei, in a process that has remained enigmatic for over a century, but which is crucial for the formation of the organelle-free zone in the center of the lens that assures clarity and function to transmit light. Nuclear degradation in lens fiber cells requires the nuclease DNase IIβ (DLAD) but the mechanism by which DLAD gains access to nuclear DNA remains unknown. In eukaryotic cells, cyclin-dependent kinase 1 (CDK1), in combination with either activator cyclins A or B, stimulates mitotic entry, in part, by phosphorylating the nuclear lamin proteins leading to the disassembly of the nuclear lamina and subsequent nuclear envelope breakdown. Although most post-mitotic cells lack CDK1 and cyclins, lens fiber cells maintain these proteins. Here, we show that loss of CDK1 from the lens inhibited the phosphorylation of nuclear lamins A and C, prevented the entry of DLAD into the nucleus, and resulted in abnormal retention of nuclei. In the presence of CDK1, a single focus of the phosphonuclear mitotic apparatus is observed, but it is not focused in CDK1-deficient lenses. CDK1 deficiency inhibited mitosis, but did not prevent DNA replication, resulting in an overall reduction of lens epithelial cells, with the remaining cells possessing an abnormally large nucleus. These observations suggest that CDK1-dependent phosphorylations required for the initiation of nuclear membrane disassembly during mitosis are adapted for removal of nuclei during fiber cell differentiation. PMID:25139855

  13. Nuclear removal during terminal lens fiber cell differentiation requires CDK1 activity: appropriating mitosis-related nuclear disassembly.

    PubMed

    Chaffee, Blake R; Shang, Fu; Chang, Min-Lee; Clement, Tracy M; Eddy, Edward M; Wagner, Brad D; Nakahara, Masaki; Nagata, Shigekazu; Robinson, Michael L; Taylor, Allen

    2014-09-01

    Lens epithelial cells and early lens fiber cells contain the typical complement of intracellular organelles. However, as lens fiber cells mature they must destroy their organelles, including nuclei, in a process that has remained enigmatic for over a century, but which is crucial for the formation of the organelle-free zone in the center of the lens that assures clarity and function to transmit light. Nuclear degradation in lens fiber cells requires the nuclease DNase IIβ (DLAD) but the mechanism by which DLAD gains access to nuclear DNA remains unknown. In eukaryotic cells, cyclin-dependent kinase 1 (CDK1), in combination with either activator cyclins A or B, stimulates mitotic entry, in part, by phosphorylating the nuclear lamin proteins leading to the disassembly of the nuclear lamina and subsequent nuclear envelope breakdown. Although most post-mitotic cells lack CDK1 and cyclins, lens fiber cells maintain these proteins. Here, we show that loss of CDK1 from the lens inhibited the phosphorylation of nuclear lamins A and C, prevented the entry of DLAD into the nucleus, and resulted in abnormal retention of nuclei. In the presence of CDK1, a single focus of the phosphonuclear mitotic apparatus is observed, but it is not focused in CDK1-deficient lenses. CDK1 deficiency inhibited mitosis, but did not prevent DNA replication, resulting in an overall reduction of lens epithelial cells, with the remaining cells possessing an abnormally large nucleus. These observations suggest that CDK1-dependent phosphorylations required for the initiation of nuclear membrane disassembly during mitosis are adapted for removal of nuclei during fiber cell differentiation. © 2014. Published by The Company of Biologists Ltd.

  14. Factors affecting the development of somatic cell nuclear transfer embryos in Cattle.

    PubMed

    Akagi, Satoshi; Matsukawa, Kazutsugu; Takahashi, Seiya

    2014-01-01

    Nuclear transfer is a complex multistep procedure that includes oocyte maturation, cell cycle synchronization of donor cells, enucleation, cell fusion, oocyte activation and embryo culture. Therefore, many factors are believed to contribute to the success of embryo development following nuclear transfer. Numerous attempts to improve cloning efficiency have been conducted since the birth of the first sheep by somatic cell nuclear transfer. However, the efficiency of somatic cell cloning has remained low, and applications have been limited. In this review, we discuss some of the factors that affect the developmental ability of somatic cell nuclear transfer embryos in cattle.

  15. Factors Affecting the Development of Somatic Cell Nuclear Transfer Embryos in Cattle

    PubMed Central

    AKAGI, Satoshi; MATSUKAWA, Kazutsugu; TAKAHASHI, Seiya

    2014-01-01

    Nuclear transfer is a complex multistep procedure that includes oocyte maturation, cell cycle synchronization of donor cells, enucleation, cell fusion, oocyte activation and embryo culture. Therefore, many factors are believed to contribute to the success of embryo development following nuclear transfer. Numerous attempts to improve cloning efficiency have been conducted since the birth of the first sheep by somatic cell nuclear transfer. However, the efficiency of somatic cell cloning has remained low, and applications have been limited. In this review, we discuss some of the factors that affect the developmental ability of somatic cell nuclear transfer embryos in cattle. PMID:25341701

  16. Nuclear Membrane-Targeted Gold Nanoparticles Inhibit Cancer Cell Migration and Invasion.

    PubMed

    Ali, Moustafa R K; Wu, Yue; Ghosh, Deepraj; Do, Brian H; Chen, Kuangcai; Dawson, Michelle R; Fang, Ning; Sulchek, Todd A; El-Sayed, Mostafa A

    2017-03-27

    Most cancer patients die from metastasis. Recent studies have shown that gold nanoparticles (AuNPs) can slow down the migration/invasion speed of cancer cells and suppress metastasis. Since nuclear stiffness of the cell largely decreases cell migration, our hypothesis is that targeting AuNPs to the cell nucleus region could enhance nuclear stiffness, and therefore inhibit cell migration and invasion. Our results showed that upon nuclear targeting of AuNPs, the ovarian cancer cell motilities decrease significantly, compared with nontargeted AuNPs. Furthermore, using atomic force microscopy, we observed an enhanced cell nuclear stiffness. In order to understand the mechanism of cancer cell migration/invasion inhibition, the exact locations of the targeted AuNPs were clearly imaged using a high-resolution three-dimensional imaging microscope, which showed that the AuNPs were trapped at the nuclear membrane. In addition, we observed a greatly increased expression level of lamin A/C protein, which is located in the inner nuclear membrane and functions as a structural component of the nuclear lamina to enhance nuclear stiffness. We propose that the AuNPs that are trapped at the nuclear membrane both (1) add to the mechanical stiffness of the nucleus and (2) stimulate the overexpression of lamin A/C located around the nuclear membrane, thus increasing nuclear stiffness and slowing cancer cell migration and invasion.

  17. Cloned ferrets produced by somatic cell nuclear transfer

    PubMed Central

    Li, Ziyi; Sun, Xingshen; Chen, Juan; Liu, Xiaoming; Wisely, Samantha M.; Zhou, Qi; Renard, Jean-Paul; Leno, Gregory H.; Engelhardt, John F.

    2007-01-01

    Somatic cell nuclear transfer (SCNT) offers great potential for developing better animal models of human disease. The domestic ferret (Mustela putorius furo) is an ideal animal model for influenza infections and potentially other human respiratory diseases such as cystic fibrosis, where mouse models have failed to reproduce the human disease phenotype. Here, we report the successful production of live cloned, reproductively competent, ferrets using species-specific SCNT methodologies. Critical to developing a successful SCNT protocol for the ferret was the finding that hormonal treatment, normally used for superovulation, adversely affected the developmental potential of recipient oocytes. The onset of Oct4 expression was delayed and incomplete in parthenogenetically activated oocytes collected from hormone-treated females relative to oocytes collected from females naturally mated with vasectomized males. Stimulation induced by mating and in vitro oocyte maturation produced the optimal oocyte recipient for SCNT. Although nuclear injection and cell fusion produced mid-term fetuses at equivalent rates (~3–4%), only cell fusion gave rise to healthy surviving clones. Single cell fusion rates and the efficiency of SCNT were also enhanced by placing two somatic cells into the perivitelline space. These species-specific modifications facilitated the birth of live, healthy, and fertile cloned ferrets. The development of microsatellite genotyping for domestic ferrets confirmed that ferret clones were genetically derived from their respective somatic cells and unrelated to their surrogate mother. With this technology, it is now feasible to begin generating genetically defined ferrets for studying transmissible and inherited human lung diseases. Cloning of the domestic ferret may also aid in recovery and conservation of the endangered black-footed ferret and European mink. PMID:16584722

  18. Role of nuclear receptors in breast cancer stem cells

    PubMed Central

    Papi, Alessio; Orlandi, Marina

    2016-01-01

    The recapitulation of primary tumour heterogenity and the existence of a minor sub-population of cancer cells, capable of initiating tumour growth in xenografts on serial passages, led to the hypothesis that cancer stem cells (CSCs) exist. CSCs are present in many tumours, among which is breast cancer. Breast CSCs (BCSCs) are likely to sustain the growth of the primary tumour mass, as well as to be responsible for disease relapse and metastatic spreading. Consequently, BCSCs represent the most significant target for new drugs in breast cancer therapy. Both the hypoxic condition in BCSCs biology and pro-inflammatory cytokine network has gained increasing importance in the recent past. Breast stromal cells are crucial components of the tumours milieu and are a major source of inflammatory mediators. Recently, the anti-inflammatory role of some nuclear receptors ligands has emerged in several diseases, including breast cancer. Therefore, the use of nuclear receptors ligands may be a valid strategy to inhibit BCSCs viability and consequently breast cancer growth and disease relapse. PMID:27022437

  19. Nuclear anomalies in exfoliated buccal cells in Pakistani cotton weavers.

    PubMed

    Khan, Abdul Wali; Nersesyan, Armen; Knasmüller, Siegfried; Moshammer, Hanns; Kundi, Michael

    2015-09-01

    Cotton workers in small weaving household factories (power looms) in Pakistan are typically exposed to high levels of cotton dusts. Working in the textile manufacturing industry has been classified as a possible human carcinogen (group 2B) by the International Agency for Research on Cancer. The study set out to determine potential cytotoxic and genotoxic effects of occupational exposure to cotton dusts in exfoliated buccal cells of exposed cotton workers. Nuclear anomalies reflecting cytotoxic and genotoxic effects were evaluated in a representative sample of 51 exposed male cotton weavers and in the same number of age-matched male non-exposed subjects applying the micronucleus cytome assay. Nuclear anomalies reflecting cytotoxicity (karyolysis, karyorrhexis, condensed chromatin and pyknosis) were significantly elevated in exposed cotton workers. The frequency of micronucleated cells increased significantly with increasing years of work in power looms (odds ratio = 1.043 per year; 95% confidence interval: 1.012-1.076, P = 0.007). Results were consistent with the typical inflammatory pattern and injury in epithelia due to unprotected occupational exposure to cotton dusts and other toxic, allergic and infectious substances in the working areas of the cotton industry. Occupational exposure in power looms induces cytotoxic effects and, upon chronic exposure, DNA damage. This may eventually result in typical obstructive patterns of pulmonary symptoms and in a clinical condition called byssinosis in exposed cotton workers. Long exposure may lead to chronic inflammation and cumulative damage of DNA in buccal stem cells that may indicate an increased risk of oropharyngeal cancer.

  20. Gnotobiotic Miniature Pig Interbreed Somatic Cell Nuclear Transfer for Xenotransplantation.

    PubMed

    Hwang, Jeong Ho; Kim, Sang Eun; Gupta, Mukesh Kumar; Lee, HoonTaek

    2016-08-01

    Transgenic animal producing technology has improved consistently over the last couple of decades. Among the available methods, somatic cell nuclear transfer (SCNT) technology was officially the most popular. However, SCNT has low efficiency and requires a highly skilled individual. Additionally, the allo-SCNT nuclear reprogramming mechanism is poorly understood in the gnotobiotic miniature pig, which is a candidate for xenotransplantation, making sampling in oocytes very difficult compared to commercial hybrid pigs. Therefore, interbreed SCNT (ibSCNT), which is a combination of miniature pig and commercial pig (Landrace based), was analyzed and was found to be similar to SCNT in terms of the rate of blastocyst formation (12.6% ± 2.9% vs. 15.5% ± 2.2%; p > 0.05). However, a significantly lower fusion rate was observed in the ibSCNT compared to normal SCNT with Landrace pig somatic cells (29.6% ± 0.8% vs. 65.0% ± 4.9%). Thus, the optimization of fusion parameters was necessary for efficient SCNT. Our results further revealed that ibSCNT by the whole-cell intracytoplasmic injection (WCICI) method had a significantly higher blastocyst forming efficiency than the electrofusion method (31.1 ± 8.5 vs. 15.5% ± 2.2%). The nuclear remodeling and the pattern of changes in acetylation at H3K9 residue were similar in both SCNT and ibSCNT embryos.

  1. Hot-compress: A new postdeposition treatment for ZnO-based flexible dye-sensitized solar cells

    SciTech Connect

    Haque Choudhury, Mohammad Shamimul

    2016-08-15

    Highlights: • A new postdeposition treatment named hot-compress is introduced. • Hot-compression gives homogeneous compact layer ZnO photoanode. • I-V and EIS analysis data confirms the efficacy of this method. • Charge transport resistance was reduced by the application of hot-compression. - Abstract: This article introduces a new postdeposition treatment named hot-compress for flexible zinc oxide–base dye-sensitized solar cells. This postdeposition treatment includes the application of compression pressure at an elevated temperature. The optimum compression pressure of 130 Ma at an optimum compression temperature of 70 °C heating gives better photovoltaic performance compared to the conventional cells. The aptness of this method was confirmed by investigating scanning electron microscopy image, X-ray diffraction, current-voltage and electrochemical impedance spectroscopy analysis of the prepared cells. Proper heating during compression lowers the charge transport resistance, longer the electron lifetime of the device. As a result, the overall power conversion efficiency of the device was improved about 45% compared to the conventional room temperature compressed cell.

  2. Characterization of Aes nuclear foci in colorectal cancer cells

    PubMed Central

    Itatani, Yoshiro; Sonoshita, Masahiro; Kakizaki, Fumihiko; Okawa, Katsuya; Stifani, Stefano; Itoh, Hideaki; Sakai, Yoshiharu; Taketo, M. Mark

    2016-01-01

    Amino-terminal enhancer of split (Aes) is a member of Groucho/Transducin-like enhancer (TLE) family. Aes is a recently found metastasis suppressor of colorectal cancer (CRC) that inhibits Notch signalling, and forms nuclear foci together with TLE1. Although some Notch-associated proteins are known to form subnuclear bodies, little is known regarding the dynamics or functions of these structures. Here, we show that Aes nuclear foci in CRC observed under an electron microscope are in a rather amorphous structure, lacking surrounding membrane. Investigation of their behaviour during the cell cycle by time-lapse cinematography showed that Aes nuclear foci dissolve during mitosis and reassemble after completion of cytokinesis. We have also found that heat shock cognate 70 (HSC70) is an essential component of Aes foci. Pharmacological inhibition of the HSC70 ATPase activity with VER155008 reduces Aes focus formation. These results provide insight into the understanding of Aes-mediated inhibition of Notch signalling. PMID:26229111

  3. Beta Cell Replacement in Mice Using Human Type 1 Diabetes Nuclear Transfer Embryonic Stem Cells.

    PubMed

    Sui, Lina; Danzl, Nichole; Campbell, Sean R; Viola, Ryan; Williams, Damian; Xing, Yuan; Wang, Yong; Phillips, Neil; Poffenberger, Greg; Johannesson, Bjarki; Oberholzer, Jose; Powers, Alvin C; Leibel, Rudolph L; Chen, Xiaojuan; Sykes, Megan; Egli, Dieter

    2017-09-20

    Beta cells derived from stem cells hold great promise for cell replacement therapy for diabetes. Here we examine the ability of nuclear transfer embryonic stem cells (NT-ES) derived from a type 1 diabetes patient to differentiate into beta cells, and provide a source of autologous islets for cell replacement. NT-ES cells differentiate in vitro with an average efficiency of 55% into C-peptide-positive cells, expressing markers of mature beta cells, including MAFA and NKX6.1. Upon transplantation in immunodeficient mice, grafted cells form vascularized islet-like structures containing MAFA/C-peptide-positive cells. These beta cells adapt insulin secretion to ambient metabolite status and show normal insulin processing. Importantly, NT-ES-beta cells maintain normal blood glucose levels after ablation of the mouse's endogenous beta cells. Cystic structures, but no teratomas, were observed in NT-ES-beta cell grafts. Isogenic induced pluripotent stem cell lines showed greater variability in beta cell differentiation. Even though different methods of somatic cell reprogramming result in stem cell lines that are molecularly indistinguishable, full differentiation competence is more common in ES cell lines than in iPS cell lines. These results demonstrate the suitability of NT-ES-beta for cell replacement for type 1 diabetes, and provide proof of principle for therapeutic cloning combined with cell therapy. © 2017 by the American Diabetes Association.

  4. Nuclear import of glucokinase in pancreatic beta-cells is mediated by a nuclear localization signal and modulated by SUMOylation.

    PubMed

    Johansson, Bente Berg; Fjeld, Karianne; Solheim, Marie Holm; Shirakawa, Jun; Zhang, Enming; Keindl, Magdalena; Hu, Jiang; Lindqvist, Andreas; Døskeland, Anne; Mellgren, Gunnar; Flatmark, Torgeir; Njølstad, Pål Rasmus; Kulkarni, Rohit N; Wierup, Nils; Aukrust, Ingvild; Bjørkhaug, Lise

    2017-10-15

    The localization of glucokinase in pancreatic beta-cell nuclei is a controversial issue. Although previous reports suggest such a localization, the mechanism for its import has so far not been identified. Using immunofluorescence, subcellular fractionation and mass spectrometry, we present evidence in support of glucokinase localization in beta-cell nuclei of human and mouse pancreatic sections, as well as in human and mouse isolated islets, and murine MIN6 cells. We have identified a conserved, seven-residue nuclear localization signal ((30)LKKVMRR(36)) in the human enzyme. Substituting the residues KK(31,32) and RR(35,36) with AA led to a loss of its nuclear localization in transfected cells. Furthermore, our data indicates that SUMOylation of glucokinase modulates its nuclear import, while high glucose concentrations do not significantly alter the enzyme nuclear/cytosolic ratio. Thus, for the first time, we provide data in support of a nuclear import of glucokinase mediated by a redundant mechanism, involving a nuclear localization signal, and which is modulated by its SUMOylation. These findings add new knowledge to the functional role of glucokinase in the pancreatic beta-cell. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Very thin and stable thin-film silicon alloy triple junction solar cells by hot wire chemical vapor deposition

    NASA Astrophysics Data System (ADS)

    Veldhuizen, L. W.; Schropp, R. E. I.

    2016-08-01

    We present a silicon-based triple junction solar cell that requires a deposition time of less than 15 min for all photoactive layers. As a low-bandgap material, we used thin layers of hydrogenated amorphous silicon germanium with lower band gap than commonly used, which is possible due to the application of hot wire chemical vapor deposition. The triple junction cell shows an initial energy conversion efficiency exceeding 10%, and with a relative performance stability within 6%, the cell shows a high tolerance to light-induced degradation. With these results, we help to demonstrate that hot wire chemical vapor deposition is a viable deposition method for the fabrication of low-cost solar cells.

  6. ERRα metabolic nuclear receptor controls growth of colon cancer cells.

    PubMed

    Bernatchez, Gérald; Giroux, Véronique; Lassalle, Thomas; Carpentier, André C; Rivard, Nathalie; Carrier, Julie C

    2013-10-01

    The estrogen-related receptor alpha (ERRα) is a nuclear receptor that acts primarily as a regulator of metabolic processes, particularly in tissues subjected to high-energy demand. In addition to its control of energy metabolism and mitochondrial biogenesis, ERRα has recently been associated with cancer progression. Notably, increased expression of ERRα has been shown in several cancerous tissues, including breast, ovary and colon. However, additional studies are required to gain insight into the action of ERRα in cancer biology, particularly in non-endocrine-related cancers. Therefore, using a short hairpin RNA-mediated approach, we investigated whether ERRα is required for the rapid growth of colon cancer cells and to maintain their neoplastic metabolic state. Results show that silencing ERRα significantly impaired colon cancer cell proliferation and colony formation in vitro as well as their in vivo tumorigenic capacity. A pronounced delay in G1-to-S cell cycle phase transition was observed in ERRα-depleted cells in association with reduced cyclin-dependent kinase 2 activity and hyperphosphorylated state of the retinoblastoma protein along with disturbed expression of several cell cycle regulators, including p15 and p27. Interestingly, ERRα-depleted HCT116 cells also displayed significant reduction in expression of a large set of key genes to glycolysis, tricarboxylic acid cycle and lipid synthesis. Furthermore, using (14)C isotope tracer analysis, ERRα depletion in colon cancer cells resulted in reduced glucose incorporation and glucose-mediated lipogenesis in these cells. These findings suggest that ERRα coordinates colon cancer cell proliferation and tumorigenic capacity with energy metabolism. Thus, ERRα could represent a promising therapeutic target in colon cancer.

  7. Transcriptomic and nuclear architecture of immune cells after LPS activation.

    PubMed

    Solinhac, Romain; Mompart, Florence; Martin, Pascal; Robelin, David; Pinton, Philippe; Iannuccelli, Eddie; Lahbib-Mansais, Yvette; Oswald, Isabelle P; Yerle-Bouissou, Martine

    2011-10-01

    Changes in the nuclear positioning of specific genes, depending on their expression status, have been observed in a large diversity of physiological processes. However, gene position is poorly documented for immune cells which have been subjected to activation following bacterial infection. Using a pig model, we focused our study on monocyte-derived macrophages and neutrophils, as they are the first lines of defence against pathogens. We examined whether changes in gene expression due to LPS activation imply that genes have repositioned in the nuclear space. We first performed a transcriptomic analysis to identify the differentially expressed genes and then analysed the networks involved during lypopolysaccharide/interferon gamma activation in monocyte-derived macrophages. This allowed us to select four up-regulated (IL1β, IL8, CXCL10 and TNFα) and four down-regulated (VIM, LGALS3, TUBA3 and IGF2) genes. Their expression statuses were verified by quantitative real-time RT-PCR before studying their behaviour in the nuclear space during macrophage activation by means of 3D fluorescence in situ hybridization. No global correlation was found between gene activity and radial positioning. Only TNFα belonging to the highly transcribed MHC region on chromosome 7 became more peripherally localized in relation to the less decondensed state of its chromosome territory (CT) in activated macrophages. The analysis of gene positioning towards their CT revealed that IL8 increases significantly its tendency to be outside its CT during the activation process. In addition, the gene to CT edge distances increase for the three up-regulated genes (IL8, CXCL10 and TNFα) among the four analysed. Contrarily, the four down-regulated genes did not change their position. The analysis of gene behaviour towards their CT was extended to include neutrophils for three (TNFα, IL8 and IL1β) up- and two (IGF2 and TUBA3) down-regulated genes, and similar results were obtained. The analysis was

  8. Route Planning and Estimate of Heat Loss of Hot Water Transportation Piping for Fuel Cell Local Energy Network

    NASA Astrophysics Data System (ADS)

    Obara, Shinya; Kudo, Kazuhiko

    The method of supplying the electric power and heat energy for the energy demand of buildings by Centralized system type and distributed system type of fuel cell network is studied. The hot-water piping route planning program of fuel cell network was developed by using genetic algorithm based on the view of TSP ( Traveling salesman problem) . In this program, the piping route planning which minimizes the quantity of heat loss in hot-water piping can be performed. The residential section model of Sapporo city of 74 buildings was analyzed, and the quantity of heat loss from the hot-water piping of both systems was estimated. Consequently, the ratio of the quantity of heat loss of a distributed system to a centralized system was about 50% in the full year average. This program is introduced into the route planning of hot- Water piping system of the fuel cell network, and plan to reduce the quantity of heat loss in a distributed system will be made.

  9. Method for somatic cell nuclear transfer in zebrafish.

    PubMed

    Siripattarapravat, K; Prukudom, S; Cibelli, J

    2016-01-01

    This chapter presents a detailed methodology for somatic cell nuclear transfer-cloning of zebrafish. We aim to place the reader in a virtual lab experience to assist acquisition of the technical skills required for reproducing the published protocol. All materials, including catalog numbers for reagents and techniques for their preparation, are provided. Our protocols describe laser inactivation of egg chromosomes, the transfer of a cell through the oocyte micropyle, and spontaneous activation of the reconstructed embryo. High-quality eggs are the key to cloning success, and Chinook salmon ovarian fluid is indispensable for keeping eggs arrested at the metaphase of meiosis II. This protocol continues to be refined by our laboratory. However, naive investigators should be able to apply it in its present form to generate cloned zebrafish. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Nuclear criticality safety evaluation -- DWPF Late Wash Facility, Salt Process Cell and Chemical Process Cell

    SciTech Connect

    Williamson, T.G.

    1994-10-17

    The Savannah River Site (SRS) High Level Nuclear Waste will be vitrified in the Defense Waste Processing Facility (DWPF) for long term storage and disposal. This is a nuclear criticality safety evaluation for the Late Wash Facility (LWF), the Salt Processing Cell (SPC) and the Chemical Processing Cell (CPC). of the DWPF. Waste salt solution is processed in the Tank Farm In-Tank Precipitation (ITP) process and is then further washed in the DWPF Late Wash Facility (LWF) before it is fed to the DWPF Salt Processing Cell. In the Salt Processing Cell the precipitate slurry is processed in the Precipitate Reactor (PR) and the resultant Precipitate Hydrolysis Aqueous (PHA) produce is combined with the sludge feed and frit in the DWPF Chemical Process Cell to produce a melter feed. The waste is finally immobilized in the Melt Cell. Material in the Tank Farm and the ITP and Extended Sludge processes have been shown to be safe against a nuclear criticality by others. The precipitate slurry feed from ITP and the first six batches of sludge feed are safe against a nuclear criticality and this evaluation demonstrates that the processes in the LWF, the SPC and the CPC do not alter the characteristics of the materials to compromise safety.

  11. Investigation of the Working Principle in an Optically Coupled Hot-Carrier Solar Cell Using the Relaxation-Time Model

    NASA Astrophysics Data System (ADS)

    Yang, J.; Patterson, R.; Feng, Y.; Shrestha, S.; Huang, S.; Conibeer, G.

    2015-04-01

    Strong optical coupling is a carrier extraction method for hot-carrier solar cells, where carriers are extracted as photons from an absorber material and then reabsorbed by a conventional photovoltaic (PV) cell. The design and working principle of this kind of optically coupled device is investigated using a relaxation-time model with practical operating conditions. This investigation is performed in the so-called "down-conversion" configuration, where the band gap of the absorber material is the same as that in the final photovoltaic cell. In this configuration, carrier impact ionization rates that are faster than thermalization rates by at least 50 times are needed to effectively down-convert the hot carriers' energy to the electronic band edge. Photon emission through radiative recombination must be enhanced by more than 500 times through photonic engineering in order to reduce Auger loss during carrier extraction. The strong luminescence from the PV cell feeding back to the absorber material will further limit the optical extraction in the near-field coupled structure, reducing the overall conversion efficiency to be lower than the ideal expectation reported previously. The strong similarity between the hot-carrier down-conversion and carrier multiplication also suggests the possible application of such optical extraction for multiple-exciton-generation solar cells, making it potentially a general carrier extraction approach for third-generation solar cells.

  12. Homocysteine-induced apoptosis in endothelial cells coincides with nuclear NOX2 and peri-nuclear NOX4 activity.

    PubMed

    Sipkens, Jessica A; Hahn, Nynke; van den Brand, Carlien S; Meischl, Christof; Cillessen, Saskia A G M; Smith, Desirée E C; Juffermans, Lynda J M; Musters, René J P; Roos, Dirk; Jakobs, Cornelis; Blom, Henk J; Smulders, Yvo M; Krijnen, Paul A J; Stehouwer, Coen D A; Rauwerda, Jan A; van Hinsbergh, Victor W M; Niessen, Hans W M

    2013-11-01

    Apoptosis of endothelial cells related to homocysteine (Hcy) has been reported in several studies. In this study, we evaluated whether reactive oxygen species (ROS)-producing signaling pathways contribute to Hcy-induced apoptosis induction, with specific emphasis on NADPH oxidases. Human umbilical vein endothelial cells were incubated with 0.01-2.5 mM Hcy. We determined the effect of Hcy on caspase-3 activity, annexin V positivity, intracellular NOX1, NOX2, NOX4, and p47(phox) expression and localization, nuclear nitrotyrosine accumulation, and mitochondrial membrane potential (ΔΨ m). Hcy induced caspase-3 activity and apoptosis; this effect was concentration dependent and maximal after 6-h exposure to 2.5 mM Hcy. It was accompanied by a significant increase in ΔΨ m. Cysteine was inactive on these parameters excluding a reactive thiol group effect. Hcy induced an increase in cellular NOX2, p47(phox), and NOX4, but not that of NOX1. 3D digital imaging microscopy followed by image deconvolution analysis showed nuclear accumulation of NOX2 and p47(phox) in endothelial cells exposed to Hcy, but not in control cells, which coincided with accumulation of nuclear nitrotyrosine residues. Furthermore, Hcy enhanced peri-nuclear localization of NOX4 coinciding with accumulation of peri-nuclear nitrotyrosine residues, a reflection of local ROS production. p47(phox) was also increased in the peri-nuclear region. The Hcy-induced increase in caspase-3 activity was prevented by DPI and apocynin, suggesting involvement of NOX activity. The data presented in this article reveal accumulation of nuclear NOX2 and peri-nuclear NOX4 accumulation as potential source of ROS production in Hcy-induced apoptosis in endothelial cells.

  13. Cell nuclear features for classification from fluorescence images

    NASA Astrophysics Data System (ADS)

    Heynen, Susanne; Hunter, Edward; Price, Jeffrey H.

    2000-04-01

    In clinical cytology, nuclear features play an important role in cell and tissue classification. To increase efficiency and decrease subjectivity of cytological results, automation of the analytic process has been proposed and discussed by many authors. This automation can be achieved by estimating the probability of occurrence of a certain class given particular features of a microscope specimen. In this paper, feature sets that might be used as inputs for mathematical cytological classification algorithms are reviewer. The primary goal was to determine the important properties of these features sets, i.e., are there mathematically efficient features that provide a more or less compete description of the cell. Under what conditions will these feature then result in optimal classification of the cells using quantitative fluorescence staining. And how would these mathematical features relate to conventional features that a human observer understands. Example human observer features are size, shape, and chromaticity o the cell nucleus while example mathematical features are image moments. If the cell image can be completely reconstructed from the feature set, then it should be possible to derive the conventional features used by human observers from the mathematical feature set for presentation to clinicians. Finally, the suitability of different mathematical decision making algorithms like probabilistic reasoning, clustering or neural networks are also briefly evaluated in the context of a mathematically complete feature set.

  14. Vertical nanopillars for in situ probing of nuclear mechanics in adherent cells

    NASA Astrophysics Data System (ADS)

    Hanson, Lindsey; Zhao, Wenting; Lou, Hsin-Ya; Lin, Ziliang Carter; Lee, Seok Woo; Chowdary, Praveen; Cui, Yi; Cui, Bianxiao

    2015-06-01

    The mechanical stability and deformability of the cell nucleus are crucial to many biological processes, including migration, proliferation and polarization. In vivo, the cell nucleus is frequently subjected to deformation on a variety of length and time scales, but current techniques for studying nuclear mechanics do not provide access to subnuclear deformation in live functioning cells. Here we introduce arrays of vertical nanopillars as a new method for the in situ study of nuclear deformability and the mechanical coupling between the cell membrane and the nucleus in live cells. Our measurements show that nanopillar-induced nuclear deformation is determined by nuclear stiffness, as well as opposing effects from actin and intermediate filaments. Furthermore, the depth, width and curvature of nuclear deformation can be controlled by varying the geometry of the nanopillar array. Overall, vertical nanopillar arrays constitute a novel approach for non-invasive, subcellular perturbation of nuclear mechanics and mechanotransduction in live cells.

  15. Vertical nanopillars for in situ probing of nuclear mechanics in adherent cells

    PubMed Central

    Hanson, Lindsey; Zhao, Wenting; Lou, Hsin-Ya; Lin, Ziliang Carter; Lee, Seok Woo; Chowdary, Praveen; Cui, Yi; Cui, Bianxiao

    2016-01-01

    The mechanical stability and deformability of the cell nucleus are crucial to many biological processes, including migration, proliferation and polarization. In vivo, the cell nucleus is frequently subjected to deformation on a variety of length and time scales, but current techniques for studying nuclear mechanics do not provide access to subnuclear deformation in live functioning cells. Here we introduce arrays of vertical nanopillars as a new method for the in situ study of nuclear deformability and the mechanical coupling between the cell membrane and the nucleus in live cells. Our measurements show that nanopillar-induced nuclear deformation is determined by nuclear stiffness, as well as opposing effects from actin and intermediate filaments. Furthermore, the depth, width and curvature of nuclear deformation can be controlled by varying the geometry of the nanopillar array. Overall, vertical nanopillar arrays constitute a novel approach for non-invasive, subcellular perturbation of nuclear mechanics and mechanotransduction in live cells. PMID:25984833

  16. Vertical nanopillars for in situ probing of nuclear mechanics in adherent cells.

    PubMed

    Hanson, Lindsey; Zhao, Wenting; Lou, Hsin-Ya; Lin, Ziliang Carter; Lee, Seok Woo; Chowdary, Praveen; Cui, Yi; Cui, Bianxiao

    2015-06-01

    The mechanical stability and deformability of the cell nucleus are crucial to many biological processes, including migration, proliferation and polarization. In vivo, the cell nucleus is frequently subjected to deformation on a variety of length and time scales, but current techniques for studying nuclear mechanics do not provide access to subnuclear deformation in live functioning cells. Here we introduce arrays of vertical nanopillars as a new method for the in situ study of nuclear deformability and the mechanical coupling between the cell membrane and the nucleus in live cells. Our measurements show that nanopillar-induced nuclear deformation is determined by nuclear stiffness, as well as opposing effects from actin and intermediate filaments. Furthermore, the depth, width and curvature of nuclear deformation can be controlled by varying the geometry of the nanopillar array. Overall, vertical nanopillar arrays constitute a novel approach for non-invasive, subcellular perturbation of nuclear mechanics and mechanotransduction in live cells.

  17. Method and apparatus for fabricating a thin-film solar cell utilizing a hot wire chemical vapor deposition technique

    DOEpatents

    Wang, Qi; Iwaniczko, Eugene

    2006-10-17

    A thin-film solar cell is provided. The thin-film solar cell comprises an a-SiGe:H (1.6 eV) n-i-p solar cell having a deposition rate of at least ten (10) .ANG./second for the a-SiGe:H intrinsic layer by hot wire chemical vapor deposition. A method for fabricating a thin film solar cell is also provided. The method comprises depositing a n-i-p layer at a deposition rate of at least ten (10) .ANG./second for the a-SiGe:H intrinsic layer.

  18. Somatic cell nuclear transfer-derived embryonic stem cell lines in humans: pros and cons.

    PubMed

    Langerova, Alena; Fulka, Helena; Fulka, Josef

    2013-12-01

    The recent paper, published by Mitalipov's group in Cell (Tachibana et al., 2013 ), reporting the production of human somatic cell nuclear transfer (SCNT) embryonic stem cells (ESCs), opens again the debate if, in the era of induced pluripotent stem cells (iPSCs), the production of these cells is indeed necessary and, if so, whether they are different from ESCs produced from spare embryos and iPSCs. It is our opinion that these questions are very difficult to answer because it is still unclear whether and how normal ESCs differ from iPSCs.

  19. Small Nuclear-powered Hot Air Balloons for the Exploration of the Deep Atmosphere of Uranus and Neptune

    NASA Astrophysics Data System (ADS)

    Van Cleve, J. E.; Grillmair, C. J.

    2001-01-01

    The Galileo probe gathered data in the Jovian atmosphere for about one hour before its destruction. For a wider perceptive on the atmospheres of the outer planets, multiple, long-lived observations platforms would be useful. In this paper we examine the basic physics of hot-air ballooning in a hydrogen atmosphere, using plutonium RTGs as a heat source. We find that such balloons are buoyant at a sufficiently great depth in these atmospheres, and derive equations for the balloon radius and mass of plutonium required as a function of atmospheric mass density and balloon material parameters. We solve for the buoyancy depth given the constraint that each probe may contain 1.0 kg of Pu, and find that the temperature at that depth is too great for conventional electronics (>70 C) for Jupiter and Saturn. However, the Pu mass constraint and the operating temperature constraint are consistent for Uranus and Neptune, and this concept may be applicable to those planets. Additional information is contained in the original extended abstract.

  20. Cytoplasmic proliferating cell nuclear antigen connects glycolysis and cell survival in acute myeloid leukemia

    PubMed Central

    Ohayon, Delphine; De Chiara, Alessia; Chapuis, Nicolas; Candalh, Céline; Mocek, Julie; Ribeil, Jean-Antoine; Haddaoui, Lamya; Ifrah, Norbert; Hermine, Olivier; Bouillaud, Frédéric; Frachet, Philippe; Bouscary, Didier; Witko-Sarsat, Véronique

    2016-01-01

    Cytosolic proliferating cell nuclear antigen (PCNA), a scaffolding protein involved in DNA replication, has been described as a key element in survival of mature neutrophil granulocytes, which are non-proliferating cells. Herein, we demonstrated an active export of PCNA involved in cell survival and chemotherapy resistance. Notably, daunorubicin-resistant HL-60 cells (HL-60R) have a prominent cytosolic PCNA localization due to increased nuclear export compared to daunorubicin-sensitive HL-60 cells (HL-60S). By interacting with nicotinamide phosphoribosyltransferase (NAMPT), a protein involved in NAD biosynthesis, PCNA coordinates glycolysis and survival, especially in HL-60R cells. These cells showed a dramatic increase in intracellular NAD+ concentration as well as glycolysis including increased expression and activity of hexokinase 1 and increased lactate production. Furthermore, this functional activity of cytoplasmic PCNA was also demonstrated in patients with acute myeloid leukemia (AML). Our data uncover a novel pathway of nuclear export of PCNA that drives cell survival by increasing metabolism flux. PMID:27759041

  1. Nuclear Localization of Diacylglycerol Kinase Alpha in K562 Cells Is Involved in Cell Cycle Progression.

    PubMed

    Poli, Alessandro; Fiume, Roberta; Baldanzi, Gianluca; Capello, Daniela; Ratti, Stefano; Gesi, Marco; Manzoli, Lucia; Graziani, Andrea; Suh, Pann-Ghill; Cocco, Lucio; Follo, Matilde Y

    2017-09-01

    Phosphatidylinositol (PI) signaling is an essential regulator of cell motility and proliferation. A portion of PI metabolism and signaling takes place in the nuclear compartment of eukaryotic cells, where an array of kinases and phosphatases localize and modulate PI. Among these, Diacylglycerol Kinases (DGKs) are a class of phosphotransferases that phosphorylate diacylglycerol and induce the synthesis of phosphatidic acid. Nuclear DGKalpha modulates cell cycle progression, and its activity or expression can lead to changes in the phosphorylated status of the Retinoblastoma protein, thus, impairing G1/S transition and, subsequently, inducing cell cycle arrest, which is often uncoupled with apoptosis or autophagy induction. Here we report for the first time not only that the DGKalpha isoform is highly expressed in the nuclei of human erythroleukemia cell line K562, but also that its nuclear activity drives K562 cells through the G1/S transition during cell cycle progression. J. Cell. Physiol. 232: 2550-2557, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  2. Identification of nuclear. tau. isoforms in human neuroblastoma cells

    SciTech Connect

    Loomis, P.A.; Howard, T.H.; Castleberry, R.P.; Binder, L.I. )

    1990-11-01

    The {tau} proteins have been reported only in association with microtubules and with ribosomes in situ, in the normal central nervous system. In addition, {tau} has been shown to be an integral component of paired helical filaments, the principal constituent of the neurofibrillary tangles found in brains of patients with Alzheimer's disease and of most aged individuals with Down syndrome (trisomy 21). The authors report here the localization of the well-characterized Tau-1 monoclonal antibody to the nucleolar organizer regions of the acrocentric chromosomes and to their interphase counterpart, the fibrillar component of the nucleolus, in human neuroblastoma cells. Similar localization to the nucleolar organizer regions was also observed in other human cell lines and in one monkey kidney cell line but was not seen in non-primate species. Immunochemically, they further demonstrated the existence of the entire {tau} molecule in the isolated nuclei of neuroblastoma cells. Nuclear {tau} proteins, like the {tau} proteins of the paired helical filaments, cannot be extracted in standard SDS-containing electrophoresis sample buffer but require pretreatment with formic acid prior to immunoblot analysis. This work indicates that {tau} may function in processes not directly associated with microtubules and that highly insoluble complexes of {tau} may also play a role in normal cellular physiology.

  3. The nuclear microprobe: An insight of applications in cell biology

    NASA Astrophysics Data System (ADS)

    Moretto, Ph.; Llabador, Y.

    1997-07-01

    During the last five years, the evolution of biomedical research based upon nuclear microprobe analysis has followed the development of experimental models of cultured or isolated cells. Fundamental studies of cellular mechanisms have been approached by means of in vitro assays associated with single cell analysis. Within those groups which are involved in such programs, special emphasis has been placed on cell culture and processing techniques which fulfill the methodological requirements for intracellular ion beam analysis. Great efforts have been orientated towards the improvement of normalization procedures. It is now possible to provide reliable quantitative results expressed in such units that they can be easily cross-checked using conventional methods. Imaging techniques have been also developed for the identification of the analyzed structures. In this paper, different domains of cell biology which have been addressed during the last years are reviewed. Studies dealing with cellular physiology and pharmacology are briefly presented as are also those related to the role of trace elements. Topics under development in our group as well as ongoing investigations will be also evoked.

  4. ASSESSMENT OF THE RADIONUCLIDE COMPOSITION OF "HOT PARTICLES" SAMPLED IN THE CHERNOBYL NUCLEAR POWER PLANT FOURTH REACTOR UNIT

    SciTech Connect

    Farfan, E.; Jannik, T.; Marra, J.

    2011-10-01

    Fuel-containing materials sampled from within the Chernobyl Nuclear Power Plant (ChNPP) 4th Reactor Unit Confinement Shelter were spectroscopically studied for gamma and alpha content. Isotopic ratios for cesium, europium, plutonium, americium, and curium were identified and the fuel burnup in these samples was determined. A systematic deviation in the burnup values based on the cesium isotopes, in comparison with other radionuclides, was observed. The conducted studies were the first ever performed to demonstrate the presence of significant quantities of {sup 242}Cm and {sup 243}Cm. It was determined that there was a systematic underestimation of activities of transuranic radionuclides in fuel samples from inside of the ChNPP Confinement Shelter, starting from {sup 241}Am (and going higher), in comparison with the theoretical calculations.

  5. Assessment of the radionuclide composition of "hot particles" sampled in the Chernobyl nuclear power plant fourth reactor unit.

    PubMed

    Bondarkov, Mikhail D; Zheltonozhsky, Viktor A; Zheltonozhskaya, Maryna V; Kulich, Nadezhda V; Maksimenko, Andrey M; Farfán, Eduardo B; Jannik, G Timothy; Marra, James C

    2011-10-01

    Fuel-containing materials sampled from within the Chernobyl Nuclear Power Plant (ChNPP) Unit 4 Confinement Shelter were spectroscopically studied for gamma and alpha content. Isotopic ratios for cesium, europium, plutonium, americium, and curium were identified, and the fuel burn-up in these samples was determined. A systematic deviation in the burn-up values based on the cesium isotopes in comparison with other radionuclides was observed. The studies conducted were the first ever performed to demonstrate the presence of significant quantities of 242Cm and 243Cm. It was determined that there was a systematic underestimation of activities of transuranic radionuclides in fuel samples from inside of the ChNPP Confinement Shelter, starting from 241Am (and going higher) in comparison with the theoretical calculations.

  6. Resonant tunneling diodes as energy-selective contacts used in hot-carrier solar cells

    NASA Astrophysics Data System (ADS)

    Takeda, Yasuhiko; Ichiki, Akihisa; Kusano, Yuya; Sugimoto, Noriaki; Motohiro, Tomoyoshi

    2015-09-01

    Among the four features unique to hot-carrier solar cells (HC-SCs): (i) carrier thermalization time and (ii) carrier equilibration time in the absorber, (iii) energy-selection width and (iv) conductance of the energy-selective contacts (ESCs), requisites of (i)-(iii) for high conversion efficiency have been clarified. We have tackled the remaining issues related to (iv) in the present study. The detailed balance model of HC-SC operation has been improved to involve a finite value of the ESC conductance to find the required values, which in turn has been revealed to be feasible using resonant tunneling diodes (RTDs) consisting of semiconductor quantum dots (QDs) and quantum wells (QWs) by means of a formulation to calculate the conductance of the QD- and QW-RTDs derived using the rigorous solutions of the effective-mass Hamiltonians. Thus, all of the four requisites unique to HC-SCs to achieve high conversion efficiency have been elucidated, and the two requisites related to the ESCs can be fulfilled using the QD- and QW-RTDs.

  7. Resonant tunneling diodes as energy-selective contacts used in hot-carrier solar cells

    SciTech Connect

    Takeda, Yasuhiko Sugimoto, Noriaki; Ichiki, Akihisa; Kusano, Yuya; Motohiro, Tomoyoshi

    2015-09-28

    Among the four features unique to hot-carrier solar cells (HC-SCs): (i) carrier thermalization time and (ii) carrier equilibration time in the absorber, (iii) energy-selection width and (iv) conductance of the energy-selective contacts (ESCs), requisites of (i)-(iii) for high conversion efficiency have been clarified. We have tackled the remaining issues related to (iv) in the present study. The detailed balance model of HC-SC operation has been improved to involve a finite value of the ESC conductance to find the required values, which in turn has been revealed to be feasible using resonant tunneling diodes (RTDs) consisting of semiconductor quantum dots (QDs) and quantum wells (QWs) by means of a formulation to calculate the conductance of the QD- and QW-RTDs derived using the rigorous solutions of the effective-mass Hamiltonians. Thus, all of the four requisites unique to HC-SCs to achieve high conversion efficiency have been elucidated, and the two requisites related to the ESCs can be fulfilled using the QD- and QW-RTDs.

  8. Development of remote crane system for use inside small argon hot-cell

    SciTech Connect

    Lee, Jong Kwang; Park, Byung Suk; Yu, Seung-Nam; Kim, Kiho; Cho, Ilje

    2013-07-01

    In this paper, we describe the design of a novel crane system for the use in a small argon hot-cell where only a pair of master-slave manipulators (MSM) is available for the remote maintenance of the crane. To increase the remote maintainability in the space-limited environment, we devised a remote actuation mechanism in which electrical parts consisting of a servo-motor, a position sensor, and two limit switches located inside the workspace of the MSM transmit power to the mechanical parts located in the ceiling. Even though the design concept does not provide thoroughly sufficient solution because the mechanical parts are placed out of the MSM's workspace, the durability of mechanical parts can be easily increased if they have a high safety margin. Therefore, the concept may be one of the best solutions for our special crane system. In addition, we developed a servo-control system based on absolute positioning technology; therefore, it is possible for us to perform the given tasks more safely through an automatic operation. (authors)

  9. Hot Cell Liners Category of Transuranic Waste Stored Below Ground within Area G

    SciTech Connect

    Jones, Robert Wesley; Hargis, Kenneth Marshall

    2014-09-01

    A large wildfire called the Las Conchas Fire burned large areas near Los Alamos National Laboratory (LANL) in 2011 and heightened public concern and news media attention over transuranic (TRU) waste stored at LANL’s Technical Area 54 (TA-54) Area G waste management facility. The removal of TRU waste from Area G had been placed at a lower priority in budget decisions for environmental cleanup at LANL because TRU waste removal is not included in the March 2005 Compliance Order on Consent (Reference 1) that is the primary regulatory driver for environmental cleanup at LANL. The Consent Order is an agreement between LANL and the New Mexico Environment Department (NMED) that contains specific requirements and schedules for cleaning up historical contamination at the LANL site. After the Las Conchas Fire, discussions were held by the U.S. Department of Energy (DOE) with the NMED on accelerating TRU waste removal from LANL and disposing it at the Waste Isolation Pilot Plant (WIPP). This report summarizes available information on the origin, configuration, and composition of the waste containers within the Hot Cell Liners category; their physical and radiological characteristics; the results of the radioassays; and the justification to reclassify the five containers as LLW rather than TRU waste.

  10. New approaches for understanding the nuclear force balance in living, adherent cells.

    PubMed

    Neelam, Srujana; Dickinson, Richard B; Lele, Tanmay P

    2016-02-01

    Cytoskeletal forces are transmitted to the nucleus to position and shape it. Linkages mediated by the LINC (linker of nucleoskeleton and cytoskeleton) complex transfer these forces to the nuclear envelope. Nuclear position and shape can be thought to be determined by a balance of cytoskeletal forces generated by microtubule motors that shear the nuclear surface, actomyosin forces that can pull, push and shear the nucleus, and intermediate filaments that may passively resist nuclear decentering and deformation. Parsing contributions of these different forces to nuclear mechanics is a very challenging task. Here we review new approaches that can be used in living cells to probe and understand the nuclear force balance.

  11. Somatic cell nuclear transfer and derivation of embryonic stem cells in the mouse.

    PubMed

    Markoulaki, Styliani; Meissner, Alexander; Jaenisch, Rudolf

    2008-06-01

    Addressing the fundamental questions of nuclear equivalence in somatic cells has fascinated scientists for decades and has resulted in the development of somatic cell nuclear transfer (SCNT) or animal cloning. SCNT involves the transfer of the nucleus of a somatic cell into the cytoplasm of an egg whose own chromosomes have been removed. In the mouse, SCNT has not only been successfully used to address the issue of nuclear equivalence, but has been used as a model system to test the hypothesis that embryonic stem cells (ESCs) derived from NT blastocysts have the potential to correct--through genetic manipulations--degenerative diseases. This paper aims to provide a comprehensive description of SCNT in the mouse and the derivation of ESCs from blastocysts generated by this technique. SCNT is a very challenging and inefficient procedure because it is technically complex, it bypasses the normal events of gamete interactions and egg activation, and it depends on adequate reprogramming of the somatic cell nucleus in vivo. Improvements in any or all those aspects may enhance the efficiency and applicability of SCNT. ESC derivation from SCNT blastocysts, on the other hand, requires the survival of only a few successfully reprogrammed cells, which have the capacity to proliferate indefinitely in vitro, maintain correct genetic and epigenetic status, and differentiate into any cell type in the body--characteristics that are essential for transplantation therapy or any other in vivo application.

  12. Karyometry of nuclear phenotypes in cutaneous squamous cell cancer.

    PubMed

    Bartels, Peter H; Bartels, Hubert G; Alberts, David S; Yozwiak, Michael; Prasad, Anil R; Glazer, Evan S; Krouse, Robert S

    2012-02-01

    To establish the karyometric characteristics of the two main nuclear phenotypes in cutaneous squamous cell cancer (cSCC) lesions. The clinical materials comprised 75 cases of cSCC, 38 with aggressive lesions and 37 with nonaggressive lesions. High-resolution images of 100 nuclei per case were recorded. Data were partitioned into four subgroups covering the range of lesion progression. Four discriminant functions were derived to distinguish aggressive from nonaggressive lesions. The most typical nuclei from the phenotype predominant in aggressive lesions and nonaggressive lesions were separated out by thresholding on the discriminant function score axes. For these homogeneous sets of nuclei the karyometric features were computed. The nuclear populations in cSCC lesions are a very heterogeneous set. There are two axes of dispersion, along the line of lesion progression and between aggressive and nonaggressive lesions. The analysis faces the difficulty that lesions from both diagnostic categories contain nuclei of the same two phenotypes with the difference between categories consisting only of differences in proportion of the two phenotypes. The nuclei of the aggressive phenotype I and nonaggressive phenotype II have substantially different chromatin patterns and can be distinguished with > 90% correct recognition rate.

  13. Nuclear matrix and structural and functional compartmentalization of the eucaryotic cell nucleus.

    PubMed

    Razin, S V; Borunova, V V; Iarovaia, O V; Vassetzky, Y S

    2014-07-01

    Becoming popular at the end of the 20th century, the concept of the nuclear matrix implies the existence of a nuclear skeleton that organizes functional elements in the cell nucleus. This review presents a critical analysis of the results obtained in the study of nuclear matrix in the light of current views on the organization of the cell nucleus. Numerous studies of nuclear matrix have failed to provide evidence of the existence of such a structure. Moreover, the existence of a filamentous structure that supports the nuclear compartmentalization appears to be unnecessary, since this function is performed by the folded genome itself.

  14. Hot gas cleanup for molten carbonate fuel cells: A zinc reactor model

    NASA Astrophysics Data System (ADS)

    Steinfeld, G.

    1980-09-01

    Of the two near term options available for desulfurization of gasifier effluent, namely low temperature cleanup utilizing absorber/stripper technology, and hot gas cleanup utilizing metal oxides, there is a clear advantage to using hot gas cleanup. Since the MCFC will operate at 1200 F, and the gasifier effluent could be between 1200 to 1900 F, a hot gas cleanup system will require little or no change in process gas temperature, thereby contributing to a high overall system efficiency. Simulated operating characteristics to aid in system design and system simulations of gasifier/MCFC systems are described. The modeling of the ZnO reactor is presented.

  15. Cytoplasmic-Nuclear Trafficking of G1/S Cell Cycle Molecules and Adult Human β-Cell Replication

    PubMed Central

    Fiaschi-Taesch, Nathalie M.; Kleinberger, Jeffrey W.; Salim, Fatimah G.; Troxell, Ronnie; Wills, Rachel; Tanwir, Mansoor; Casinelli, Gabriella; Cox, Amy E.; Takane, Karen K.; Srinivas, Harish; Scott, Donald K.; Stewart, Andrew F.

    2013-01-01

    Harnessing control of human β-cell proliferation has proven frustratingly difficult. Most G1/S control molecules, generally presumed to be nuclear proteins in the human β-cell, are in fact constrained to the cytoplasm. Here, we asked whether G1/S molecules might traffic into and out of the cytoplasmic compartment in association with activation of cell cycle progression. Cdk6 and cyclin D3 were used to drive human β-cell proliferation and promptly translocated into the nucleus in association with proliferation. In contrast, the cell cycle inhibitors p15, p18, and p19 did not alter their location, remaining cytoplasmic. Conversely, p16, p21, and p27 increased their nuclear frequency. In contrast once again, p57 decreased its nuclear frequency. Whereas proliferating β-cells contained nuclear cyclin D3 and cdk6, proliferation generally did not occur in β-cells that contained nuclear cell cycle inhibitors, except p21. Dynamic cytoplasmic-nuclear trafficking of cdk6 was confirmed using green fluorescent protein–tagged cdk6 and live cell imaging. Thus, we provide novel working models describing the control of cell cycle progression in the human β-cell. In addition to known obstacles to β-cell proliferation, cytoplasmic-to-nuclear trafficking of G1/S molecules may represent an obstacle as well as a therapeutic opportunity for human β-cell expansion. PMID:23493571

  16. Novel nuclear hENT2 isoforms regulate cell cycle progression via controlling nucleoside transport and nuclear reservoir.

    PubMed

    Grañé-Boladeras, Natalia; Spring, Christopher M; Hanna, W J Brad; Pastor-Anglada, Marçal; Coe, Imogen R

    2016-12-01

    Nucleosides participate in many cellular processes and are the fundamental building blocks of nucleic acids. Nucleoside transporters translocate nucleosides across plasma membranes although the mechanism by which nucleos(t)ides are translocated into the nucleus during DNA replication is unknown. Here, we identify two novel functional splice variants of equilibrative nucleoside transporter 2 (ENT2), which are present at the nuclear envelope. Under proliferative conditions, these splice variants are up-regulated and recruit wild-type ENT2 to the nuclear envelope to translocate nucleosides into the nucleus for incorporation into DNA during replication. Reduced presence of hENT2 splice variants resulted in a dramatic decrease in cell proliferation and dysregulation of cell cycle due to a lower incorporation of nucleotides into DNA. Our findings support a novel model of nucleoside compartmentalisation at the nuclear envelope and translocation into the nucleus through hENT2 and its variants, which are essential for effective DNA synthesis and cell proliferation.

  17. Recent progress in bovine somatic cell nuclear transfer.

    PubMed

    Akagi, Satoshi; Geshi, Masaya; Nagai, Takashi

    2013-03-01

    Bovine somatic cell nuclear transfer (SCNT) embryos can develop to the blastocyst stage at a rate similar to that of embryos produced by in vitro fertilization. However, the full-term developmental rate of SCNT embryos is very low, owing to the high embryonic and fetal losses after embryo transfer. In addition, increased birth weight and postnatal mortality are observed at high rates in cloned calves. The low efficiency of SCNT is probably attributed to incomplete reprogramming of the donor nucleus and most of the developmental problems of clones are thought to be caused by epigenetic defects. Applications of SCNT will depend on improvement in the efficiency of production of healthy cloned calves. In this review, we discuss problems and recent progress in bovine SCNT.

  18. Global reorganization of the nuclear landscape in senescent cells.

    PubMed

    Chandra, Tamir; Ewels, Philip Andrew; Schoenfelder, Stefan; Furlan-Magaril, Mayra; Wingett, Steven William; Kirschner, Kristina; Thuret, Jean-Yves; Andrews, Simon; Fraser, Peter; Reik, Wolf

    2015-02-03

    Cellular senescence has been implicated in tumor suppression, development, and aging and is accompanied by large-scale chromatin rearrangements, forming senescence-associated heterochromatic foci (SAHF). However, how the chromatin is reorganized during SAHF formation is poorly understood. Furthermore, heterochromatin formation in senescence appears to contrast with loss of heterochromatin in Hutchinson-Gilford progeria. We mapped architectural changes in genome organization in cellular senescence using Hi-C. Unexpectedly, we find a dramatic sequence- and lamin-dependent loss of local interactions in heterochromatin. This change in local connectivity resolves the paradox of opposing chromatin changes in senescence and progeria. In addition, we observe a senescence-specific spatial clustering of heterochromatic regions, suggesting a unique second step required for SAHF formation. Comparison of embryonic stem cells (ESCs), somatic cells, and senescent cells shows a unidirectional loss in local chromatin connectivity, suggesting that senescence is an endpoint of the continuous nuclear remodelling process during differentiation. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  19. Cell penetrating peptide inhibitors of Nuclear Factor-kappa B

    PubMed Central

    Orange, J. S.; May, M. J.

    2010-01-01

    The nuclear factor kappa B (NF-κB) transcription factors are activated by a range of stimuli including pro-inflammatory cytokines. Active NF-κB regulates the expression of genes involved in inflammation and cell survival and aberrant NF-κB activity plays pathological roles in certain types of cancer and diseases characterized by chronic inflammation. NF-κB signaling is an attractive target for the development of novel anti-inflammatory or anti-cancer drugs and we discuss here how the method of peptide transduction has been used to specifically target NF-κB. Peptide transduction relies on the ability of certain small cell-penetrating peptides (CPPs) to enter cells, and a panel of CPP-linked inhibitors (CPP-Is) has been developed to directly inhibit NF-κB signaling. Remarkably, several of these NF-κB-targeting CPP-Is are effective in vivo and therefore offer exciting potential in the clinical setting. PMID:18668204

  20. Global Reorganization of the Nuclear Landscape in Senescent Cells

    PubMed Central

    Chandra, Tamir; Ewels, Philip Andrew; Schoenfelder, Stefan; Furlan-Magaril, Mayra; Wingett, Steven William; Kirschner, Kristina; Thuret, Jean-Yves; Andrews, Simon; Fraser, Peter; Reik, Wolf

    2015-01-01

    Summary Cellular senescence has been implicated in tumor suppression, development, and aging and is accompanied by large-scale chromatin rearrangements, forming senescence-associated heterochromatic foci (SAHF). However, how the chromatin is reorganized during SAHF formation is poorly understood. Furthermore, heterochromatin formation in senescence appears to contrast with loss of heterochromatin in Hutchinson-Gilford progeria. We mapped architectural changes in genome organization in cellular senescence using Hi-C. Unexpectedly, we find a dramatic sequence- and lamin-dependent loss of local interactions in heterochromatin. This change in local connectivity resolves the paradox of opposing chromatin changes in senescence and progeria. In addition, we observe a senescence-specific spatial clustering of heterochromatic regions, suggesting a unique second step required for SAHF formation. Comparison of embryonic stem cells (ESCs), somatic cells, and senescent cells shows a unidirectional loss in local chromatin connectivity, suggesting that senescence is an endpoint of the continuous nuclear remodelling process during differentiation. PMID:25640177

  1. Somatic cell nuclear transfer (cloning): implications for the medical practitioner.

    PubMed

    Tong, W F; Ng, Y F; Ng, S C

    2002-07-01

    The current century will bring tremendous changes to the science and the practice of medicine. This century will be acknowledged as the century of Biology as the fusion of molecular genetics and experimental embryology pushes the barriers of science beyond perimeters that we have thought existed, as much as the past century was the century of Physics, with all the exact scientific calculations and predictions, resulting in electricity, nuclear power and quantum physics. The first major breakthrough has been the pioneering work of Wilmut and Campbell, first with the birth of Megan and Moran in 1995 (1), followed by the birth of Dolly the sheep, the first reported mammalian clone from a fully differentiated adult cell, reported in July 1996 (2). However, current cloning techniques are an extension of over 40 years of research using nuclei derived from non-human embryonic and fetal cells. However, following the birth of Dolly, the prospects of cloning technology have extended to ethically hazier areas of human cloning and embryonic stem cell research. This review hopes to bring the reader closer to the science and the ethics of this new technology, and what the implications are for the medical practitioner.

  2. Exosomes surf on filopodia to enter cells at endocytic hot spots, traffic within endosomes, and are targeted to the ER

    PubMed Central

    Hean, Justin; Trojer, Dominic; Steib, Emmanuelle; von Bueren, Stefan; Graff-Meyer, Alexandra; Genoud, Christel; Martin, Katrin; Pizzato, Nicolas; Voshol, Johannes; Morrissey, David V.; Andaloussi, Samir E.L.; Wood, Matthew J.

    2016-01-01

    Exosomes are nanovesicles released by virtually all cells, which act as intercellular messengers by transfer of protein, lipid, and RNA cargo. Their quantitative efficiency, routes of cell uptake, and subcellular fate within recipient cells remain elusive. We quantitatively characterize exosome cell uptake, which saturates with dose and time and reaches near 100% transduction efficiency at picomolar concentrations. Highly reminiscent of pathogenic bacteria and viruses, exosomes are recruited as single vesicles to the cell body by surfing on filopodia as well as filopodia grabbing and pulling motions to reach endocytic hot spots at the filopodial base. After internalization, exosomes shuttle within endocytic vesicles to scan the endoplasmic reticulum before being sorted into the lysosome as their final intracellular destination. Our data quantify and explain the efficiency of exosome internalization by recipient cells, establish a new parallel between exosome and virus host cell interaction, and suggest unanticipated routes of subcellular cargo delivery. PMID:27114500

  3. Reaction System Design for NO2 Oxidation of Used Nuclear Fuel for the FY17 Hot Test

    SciTech Connect

    Johnson, Jared A.; DelCul, Guillermo Daniel

    2016-08-25

    A dry pretreatment process based on the oxidation of used nuclear fuel to convert it to a fine powder is being studied for the removal and capture of tritium and iodine before subsequent processing. The process converts oxide fuel into a fine powder at low temperature using NO2/O2 mixtures. The form of the powder product can be selected to be U3O8, UO3, or a nitrate by adjusting the processing conditions. All the fundamental tenets of the process have been successfully demonstrated as a proof of principle, and many aspects have been corroborated multiple times at laboratory scale. The present thrust is to develop the process to a technology-readiness level sufficient to evaluate and estimate the cost of an engineeringscale implementation. A previous roadmap analysis of the implementation determined that the most desirable approach would be based on kilogram-scale experiments using real fuel in parallel with multi- Kg testing of prototype systems using surrogate material. Following the planned approach, kilogram-scale experiments will be conducted in FY 2017. This report describes the primary reaction system designs and the equipment that has been fabricated. Details related to instrumentation, data acquisition, and controls have been previously reported.

  4. Nuclear orphan receptor TLX affects gene expression, proliferation and cell apoptosis in beta cells

    SciTech Connect

    Shi, Xiaoli; Xiong, Xiaokan; Dai, Zhe; Deng, Haohua; Sun, Li; Hu, Xuemei; Zhou, Feng; Xu, Yancheng

    2015-12-04

    Nuclear orphan receptor TLX is an essential regulator of the growth of neural stem cells. However, its exact function in pancreatic islet cells is still unknown. In the present study, gene expression profiling analysis revealed that overexpression of TLX in beta cell line MIN6 causes suppression of 176 genes and upregulation of 49 genes, including a cadre of cell cycle, cell proliferation and cell death control genes, such as Btg2, Ddit3 and Gadd45a. We next examined the effects of TLX overexpression on proliferation, apoptosis and insulin secretion in MIN6 cells. Proliferation analysis using EdU assay showed that overexpression of TLX increased percentage of EdU-positive cells. Cell cycle and apoptosis analysis revealed that overexpression of TLX in MIN6 cells resulted in higher percentage of cells exiting G1 into S-phase, and a 58.8% decrease of cell apoptosis induced by 0.5 mM palmitate. Moreover, TLX overexpression did not cause impairment of insulin secretion. Together, we conclude that TLX is among factors capable of controlling beta cell proliferation and survival, which may serve as a target for the development of novel therapies for diabetes. - Highlights: • TLX overexpression in MIN6 cell causes significant expression changes of 225 genes. • TLX overexpression promotes MIN6 cell proliferation and decreases cell apoptosis. • TLX overexpression does not cause impairment of insulin secretion.

  5. Modeling and characterization of double resonant tunneling diodes for application as energy selective contacts in hot carrier solar cells

    NASA Astrophysics Data System (ADS)

    Jehl, Zacharie; Suchet, Daniel; Julian, Anatole; Bernard, Cyril; Miyashita, Naoya; Gibelli, Francois; Okada, Yoshitaka; Guillemolles, Jean-Francois

    2017-02-01

    Double resonant tunneling barriers are considered for an application as energy selective contacts in hot carrier solar cells. Experimental symmetric and asymmetric double resonant tunneling barriers are realized by molecular beam epitaxy and characterized by temperature dependent current-voltage measurements. The negative differential resistance signal is enhanced for asymmetric heterostructures, and remains unchanged between low- and room-temperatures. Within Tsu-Esaki description of the tunnel current, this observation can be explained by the voltage dependence of the tunnel transmission amplitude, which presents a resonance under finite bias for asymmetric structures. This effect is notably discussed with respect to series resistance. Different parameters related to the electronic transmission of the structure and the influence of these parameters on the current voltage characteristic are investigated, bringing insights on critical processes to optimize in double resonant tunneling barriers applied to hot carrier solar cells.

  6. Nuclear vasohibin-2 promotes cell proliferation by inducing G0/G1 to S phase progression.

    PubMed

    Ge, Qianqian; Zhou, Jia; Tu, Min; Xue, Xiaofeng; Li, Zhanjun; Lu, Zipeng; Wei, Jishu; Song, Guoxin; Chen, Jianmin; Guo, Feng; Jiang, Kuirong; Miao, Yi; Gao, Wentao

    2015-09-01

    As a member of the vasohibin (VASH2) family, VASH2 is localized intracellularly as a nuclear and cytoplasmic type. Cytoplasmic VASH2 is associated with carcinoma angiogenesis and malignant transformation and promotes cancer growth. However, the function of nuclear VASH2 has yet to be investigated. The aim of the present study was to detect the nuclear VASH2 expression profile in human organs and tissues by protein microarray technique. To examine the function of nuclear VASH2, we analyzed the relationship between nuclear VASH2 and Ki-67, and stably constructed VASH2 overexpression and knockdown in LO2 and HepG2 cell lines, based on a previous study in hepatic cells. The study was conducted using bromodeoxyuridine, immunofluorescent staining, western blot analysis and flow cytometry. Nuclear VASH2 was highly expressed in actively dividing cells in normal and cancer tissues. There was a significant positive correlation between nuclear VASH2 and Ki-67, indicating that nuclear VASH2 positively correlated with cell proliferation in normal and cancer tissues. The bromodeoxyuridine (BrdU) proliferation test showed that nuclear VASH2 increased the S-phase population and promoted cell proliferation, while VASH2 knockdown reduced BrdU absorbance. Cell cycle analysis revealed that nuclear VASH2 overexpression increased the S-phase population in LO2 and HepG2 cells, while nuclear VASH2 knockdown reduced the S-phase population and increased the G0/G1 population. The findings of this study challenge the classic view of VASH2, which was previously reported as an angiogenesis factor. Furthermore, to the best of our knowledge, these results are the first clinical data indicating that nuclear VASH2, but not cytoplasmic VASH2, promotes cell proliferation by driving the cell cycle from the G0/G1 to S phase.

  7. Value of volume weighted mean nuclear volume in grading and prognosis of renal cell carcinoma.

    PubMed Central

    Artacho-Pérula, E; Roldán-Villalobos, R; Martínez-Cuevas, J F

    1994-01-01

    AIMS--To perform stereological quantitation of volume weighted mean nuclear volume in renal cell carcinomas; and to correlate the data obtained with recognised clinical and pathological variables and determine their prognostic value. METHODS--The point-sampled intercepts method was used to estimate mean nuclear volume in 63 cases of clear cell renal carcinoma diagnosed between 1980 and 1988. New paraffin wax embedded histological sections were analysed after systematic sampling and the test systems superimposed on a projected microscopic image to measure nuclear intercept lengths. After mathematical estimation of mean nuclear volume, statistical analyses of the data in relation to clinical and pathological variables as well as the prognostic impact were investigated. RESULTS--The mean nuclear volume was significantly associated with tumour dedifferentiation. However, mean nuclear volume showed no statistical differences with sex, age, and clinical stage. The prognostic value of mean nuclear volume, nuclear grading, and clinical stage in renal cell carcinomas was high: mean nuclear volume greater than 140 micron3 was associated with short term survival. CONCLUSIONS--Measurement of mean nuclear volume was useful as a guide to objective grading of renal cell carcinomas, though there was an overlap between tumour grades. Based on the limited number of cases analysed, the mean nuclear volume is proposed as an additional prognostic indicator. Images PMID:8027369

  8. Atypical nuclear localization of VIP receptors in glioma cell lines and patients

    SciTech Connect

    Barbarin, Alice; Séité, Paule; Godet, Julie; Bensalma, Souheyla; Muller, Jean-Marc; Chadéneau, Corinne

    2014-11-28

    Highlights: • The VIP receptor VPAC1 contains a putative NLS signal. • VPAC1 is predominantly nuclear in GBM cell lines but not VPAC2. • Non-nuclear VPAC1/2 protein expression is correlated with glioma grade. • Nuclear VPAC1 is observed in 50% of stage IV glioma (GBM). - Abstract: An increasing number of G protein-coupled receptors, like receptors for vasoactive intestinal peptide (VIP), are found in cell nucleus. As VIP receptors are involved in the regulation of glioma cell proliferation and migration, we investigated the expression and the nuclear localization of the VIP receptors VPAC1 and VPAC2 in this cancer. First, by applying Western blot and immunofluorescence detection in three human glioblastoma (GBM) cell lines, we observed a strong nuclear staining for the VPAC1 receptor and a weak nuclear VPAC2 receptor staining. Second, immunohistochemical staining of VPAC1 and VPAC2 on tissue microarrays (TMA) showed that the two receptors were expressed in normal brain and glioma tissues. Expression in the non-nuclear compartment of the two receptors significantly increased with the grade of the tumors. Analysis of nuclear staining revealed a significant increase of VPAC1 staining with glioma grade, with up to 50% of GBM displaying strong VPAC1 nuclear staining, whereas nuclear VPAC2 staining remained marginal. The increase in VPAC receptor expression with glioma grades and the enhanced nuclear localization of the VPAC1 receptors in GBM might be of importance for glioma progression.

  9. Nuclear actin is partially associated with Cajal bodies in human cells in culture and relocates to the nuclear periphery after infection of cells by adenovirus 5.

    PubMed

    Gedge, L J E; Morrison, E E; Blair, G E; Walker, J H

    2005-02-15

    Cajal bodies are intra-nuclear structures enriched in proteins involved in transcription and mRNA processing. In this study, immunofluorescence microscopy experiments using a highly specific antibody to actin revealed nuclear actin spots that colocalized in part with p80 coilin-positive Cajal bodies. Actin remained associated with Cajal bodies in cells extracted to reveal the nuclear matrix. Adenovirus infection, which is known to disassemble Cajal bodies, resulted in loss of actin from these structures late in infection. In infected cells, nuclear actin was observed to relocate to structures at the periphery of the nucleus, inside the nuclear envelope. Based on these findings, it is suggested that actin may play an important role in the organization or function of the Cajal body.

  10. Improvement of canine somatic cell nuclear transfer procedure.

    PubMed

    Jang, G; Oh, H J; Kim, M K; Fibrianto, Y H; Hossein, M S; Kim, H J; Kim, J J; Hong, S G; Park, J E; Kang, S K; Lee, B C

    2008-01-15

    The purpose of the present study on canine somatic cell nuclear transfer (SCNT) was to evaluate the effects of fusion strength, type of activation, culture media and site of transfer on developmental potential of SCNT embryos. We also examined the potential of enucleated bovine oocytes to serve as cytoplast recipients of canine somatic cells. Firstly, we evaluated the morphological characteristics of in vivo-matured canine oocytes collected by retrograde flushing of the oviducts 72 h after ovulation. Secondly, the effectiveness of three electrical strengths (1.8, 2.3 and 3.3 kV/cm), used twice for 20 micros, on fusion of canine cytoplasts with somatic cells were compared. Then, we compared: (1) chemical versus electrical activation (a) after parthenogenetic activation or (b) after reconstruction of canine oocytes with somatic cells; (2) culture of resulting intergeneric (IG) embryos in either (a) mSOF or (b) TCM-199. The exposure time to 6-DMAP was standardized by using bovine oocytes reconstructed with canine somatic cells. Bovine oocytes were used for SCNT after a 22 h in vitro maturation interval. The fusion rate was significantly higher in the 3.3 kV/cm group than in the 1.8 and 2.3 kV/cm treatment groups. After parthenogenesis or SCNT with chemical activation, 3.4 and 5.8%, respectively, of the embryos developed to the morula stage, as compared to none of the embryos produced using electrical activation. Later developmental stages (8-16 cells) were transferred to the uterine horn of eight recipients, but no pregnancy was detected. However, IG cloned embryos (bovine cytoplast/canine somatic cell) were capable of in vitro blastocyst development. In vitro developmental competence of IG cloned embryos was improved after exposure to 6-DMAP for 4 h as compared to 0, 2 or 6h exposure, although the increase was not significantly different among culture media. In summary, for production of canine SCNT embryos, we recommend fusion at 3.3 kV/cm, chemical activation

  11. Nuclear cathepsin L activity is required for cell cycle progression of colorectal carcinoma cells.

    PubMed

    Tamhane, Tripti; Lllukkumbura, Rukshala; Lu, Shiying; Maelandsmo, Gunhild M; Haugen, Mads H; Brix, Klaudia

    2016-03-01

    Prominent tasks of cysteine cathepsins involve endo-lysosomal proteolysis and turnover of extracellular matrix constituents or plasma membrane proteins for maintenance of intestinal homeostasis. Here we report on enhanced levels and altered subcellular localization of distinct cysteine cathepsins in adenocarcinoma tissue in comparison to adjacent normal colon. Immunofluorescence and immunoblotting investigations revealed the presence of cathepsin L in the nuclear compartment in addition to its expected endo-lysosomal localization in colorectal carcinoma cells. Cathepsin L was represented as the full-length protein in the nuclei of HCT116 cells from which stefin B, a potent cathepsin L inhibitor, was absent. Fluorescence activated cell sorting analyses with synchronized cell cultures revealed deceleration of cell cycle progression of HCT116 cells upon inhibition of cathepsin L activity, while expression of cathepsin L-enhanced green fluorescent protein chimeras accelerated S-phase entry. We conclude that the activity of cathepsin L is high in the nucleus of colorectal carcinoma cells because of lacking stefin B inhibitory activity. Furthermore, we hypothesize that nuclear cathepsin L accelerates cell cycle progression of HCT116 cells thereby supporting the notion that cysteine cathepsins may play significant roles in carcinogenesis due to deregulated trafficking.

  12. Quantification of nanoscale nuclear refractive index changes during the cell cycle

    NASA Astrophysics Data System (ADS)

    Bista, Rajan K.; Uttam, Shikhar; Wang, Pin; Staton, Kevin; Choi, Serah; Bakkenist, Christopher J.; Hartman, Douglas J.; Brand, Randall E.; Liu, Yang

    2011-07-01

    Intrigued by our recent finding that the nuclear refractive index is significantly increased in malignant cells and histologically normal cells in clinical histology specimens derived from cancer patients, we sought to identify potential biological mechanisms underlying the observed phenomena. The cell cycle is an ordered series of events that describes the intervals of cell growth, DNA replication, and mitosis that precede cell division. Since abnormal cell cycles and increased proliferation are characteristic of many human cancer cells, we hypothesized that the observed increase in nuclear refractive index could be related to an abundance or accumulation of cells derived from cancer patients at a specific point or phase(s) of the cell cycle. Here we show that changes in nuclear refractive index of fixed cells are seen as synchronized populations of cells that proceed through the cell cycle, and that increased nuclear refractive index is strongly correlated with increased DNA content. We therefore propose that an abundance of cells undergoing DNA replication and mitosis may explain the increase in nuclear refractive index observed in both malignant and histologically normal cells from cancer patients. Our findings suggest that nuclear refractive index may be a novel physical parameter for early cancer detection and risk stratification.

  13. Study of a ;hot; particle with a matrix of U-bearing metallic Zr: Clue to supercriticality during the Chernobyl nuclear accident

    NASA Astrophysics Data System (ADS)

    Pöml, P.; Burakov, B.

    2017-05-01

    This paper is dedicated to the 30th anniversary of the severe nuclear accident that occurred at the Chernobyl NPP on 26 April 1986. A detailed study on a Chernobyl ;hot; particle collected from contaminated soil was performed. Optical and electron microscopy, as well as quantitative x-ray microbeam analysis methods were used to determine the properties of the sample. The results show that the particle (≈ 240 x 165 μm) consists of a metallic Zr matrix containing 2-3 wt. % U and bearing veins of an U,Nb admixture. The metallic Zr matrix contains two phases with different amounts of O with the atomic proportions (U,Zr,Nb)0.73O0.27 and (U,Zr,Nb)0.61O0.39. The results confirm the interaction between UO2 fuel and zircaloy cladding in the reactor core. To explain the process of formation of the particle, its properties are compared to laboratory experiments. Because of the metallic nature of the particle it is concluded that it must have formed during a very high temperature (> 2400∘C) process that lasted for only a very short time (few microseconds or less); otherwise the particle should have been oxidised. Such a rapid very high temperature process indicates that at least part of the reactor core could have been supercritical prior to an explosion as it was previously suggested in the literature.

  14. Nuclear transition between the conjunction cells of Phaeodactylum tricornutum Bohlin (Bacillariophyta)

    NASA Astrophysics Data System (ADS)

    Li, Si; Pan, Kehou; Zhu, Baohua; Zhang, Lin

    2012-09-01

    Phaeodactylum tricornutum is one of the important marine diatoms for oceanic primary production. Its reproduction has profound significance in the life cycle; however, the nuclear behavior during its sexual reproduction was not clear. In this study, we observed the nuclear transition and determined its correlation with cell conjunction. It was found that two cells jointed at their apices first and swung and aligned each other immediately, and nucleus from one cell was able to transfer into another one during cell conjugation. The cell pairs conjugated for nuclear transition were different from those formed in mitosis in hypovalve thickness and cellular arrangement. Our findings proved the existence of sexual reproduction in P. tricornutum.

  15. Influence of red blood cell concentrations on the measurement of turbulence using hot-film anemometer.

    PubMed

    Sallam, A M; Hwang, N H

    1983-11-01

    Measurement of local velocity fluctuations was made with an L-shaped conical hot-film probe in a submerged circular jet. The experiment was carried out in solutions of washed human red blood cells (RBC) in a phosphate buffer solution (PBS), at hematocrit concentrations (Ht percent) of 10, 19, 29, and 38 percent. The viscosity of the testing solutions was kept at 3.2 c.p. by adding proper amount of dextran. The experiment was conducted at Reynolds numbers (NR) 674, 963, 1255 and 1410, based on the jet exit velocity and exit diameter. Statistical analyses were performed on the recorded instantaneous velocity signals to obtain the root-mean-square (rms) values, the probability density functions (PDF) and the power spectral density functions (PSDF) of the signals. Within the range tested, we noticed an incidental rise in rms values at 19 to 29 Ht percent for NR = 963 similar to those reported earlier in the literature. Further analyses using PDF and PSDF, however, showed neither a trend nor any physical significance of this rise. Based on the analyses of both the PDF and the PSDF, we believe that the incidental rise in rms value can be partially attributed to the high spikes registered by the probe in a high RBC concentrations fluid flow. The bombardment of RBC on the probe thermal boundary layer may cause a characteristic change in the probe response to certain flow phenomenon, at least within the Reynolds number range used in this study.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Somatic cell nuclear transfer cloning: practical applications and current legislation.

    PubMed

    Niemann, H; Lucas-Hahn, A

    2012-08-01

    Somatic cloning is emerging as a new biotechnology by which the opportunities arising from the advances in molecular genetics and genome analysis can be implemented in animal breeding. Significant improvements have been made in SCNT protocols in the past years which now allow to embarking on practical applications. The main areas of application of SCNT are: Reproductive cloning, therapeutic cloning and basic research. A great application potential of SCNT based cloning is the production of genetically modified (transgenic) animals. Somatic cell nuclear transfer based transgenic animal production has significant advances over the previously employed microinjection of foreign DNA into pronuclei of zygotes. This cell based transgenesis is compatible with gene targeting and allows both, the addition of a specific gene and the deletion of an endogenous gene. Efficient transgenic animal production provides numerous opportunities for agriculture and biomedicine. Regulatory agencies around the world have agreed that food derived from cloned animals and their offspring is safe and there is no scientific basis for questioning this. Commercial application of somatic cloning within the EU is via the Novel Food regulation EC No. 258/97. Somatic cloning raises novel questions regarding the ethical and moral status of animals and their welfare which has prompted a controversial discussion in Europe which has not yet been resolved. © 2012 Blackwell Verlag GmbH.

  17. Water Permeability of Chlorella Cell Membranes by Nuclear Magnetic Resonance

    PubMed Central

    Stout, Darryl G.; Steponkus, Peter L.; Bustard, Larry D.; Cotts, Robert M.

    1978-01-01

    Measurement by two nuclear magnetic resonance (NMR) techniques of the mean residence time τa of water molecules inside Chlorella vulgaris (Beijerinck) var. “viridis” (Chodot) is reported. The first is the Conlon and Outhred (1972 Biochim Biophys Acta 288: 354-361) technique in which extracellular water is doped with paramagnetic Mn2+ ions. Some complications in application of this technique are identified as being caused by the affinity of Chlorella cell walls for Mn2+ ions which shortens the NMR relaxation times of intra- and extracellular water. The second is based upon observations of effects of diffusion on the spin echo of intra- and extracellular water. Echo attenuation of intracellular water is distinguished from that of extracellular water by the extent to which diffusive motion is restricted. Intracellular water, being restricted to the cell volume, suffers less echo attenuation. From the dependence of echo amplitude upon gradient strength at several values of echo time, the mean residence time of intracellular water can be determined. From the mean residence time of intracellular water, the diffusional water permeability coefficient of the Chlorella membrane is calculated to be 2.1 ± 0.4 × 10−3 cm sec−1. PMID:16660456

  18. Propagation of elite rescue dogs by somatic cell nuclear transfer.

    PubMed

    Oh, Hyun Ju; Choi, Jin; Kim, Min Jung; Kim, Geon A; Jo, Young Kwang; Choi, Yoo Bin; Lee, Byeong Chun

    2016-01-01

    The objective of the present study was to compare the efficiency of two oocyte activation culture media to produce cloned dogs from an elite rescue dog and to analyze their behavioral tendencies. In somatic cell nuclear transfer procedure, fused couplets were activated by calcium ionophore treatment for 4 min, cultured in two media: modified synthetic oviduct fluid (mSOF) with 1.9 mmol/L 6-dimethylaminopyridine (DMAP) (SOF-DMAP) or porcine zygote medium (PZM-5) with 1.9 mmol/L DMAP (PZM-DMAP) for 4 h, and then were transferred into recipients. After embryo transfer, pregnancy was detected in one out of three surrogate mothers that received cloned embryos from the PZM-DMAP group (33.3%), and one pregnancy (25%) was detected in four surrogate mothers receiving cloned embryos from the SOF-DMAP group. Each pregnant dog gave birth to one healthy cloned puppy by cesarean section. We conducted the puppy aptitude test with two cloned puppies; the two cloned puppies were classified as the same type, accepting humans and leaders easily. The present study indicated that the type of medium used in 6-DMAP culture did not increase in cloning efficiency and dogs cloned using donor cells derived from one elite dog have similar behavioral tendencies.

  19. Concise Review: Embryonic Stem Cells Derived by Somatic Cell Nuclear Transfer: A Horse in the Race?

    PubMed

    Wolf, Don P; Morey, Robert; Kang, Eunju; Ma, Hong; Hayama, Tomonari; Laurent, Louise C; Mitalipov, Shoukhrat

    2017-01-01

    Embryonic stem cells (ESC) hold promise for the treatment of human medical conditions but are allogeneic. Here, we consider the differences between autologous pluripotent stem cells produced by nuclear transfer (NT-ESCs) and transcription factor-mediated, induced pluripotent stem cells (iPSCs) that impact the desirability of each of these cell types for clinical use. The derivation of NT-ESCs is more cumbersome and requires donor oocytes; however, the use of oocyte cytoplasm as the source of reprogramming factors is linked to a key advantage of NT-ESCs-the ability to replace mutant mitochondrial DNA in a patient cell (due to either age or inherited disease) with healthy donor mitochondria from an oocyte. Moreover, in epigenomic and transcriptomic comparisons between isogenic iPSCs and NT-ESCs, the latter produced cells that more closely resemble bona fide ESCs derived from fertilized embryos. Thus, although NT-ESCs are more difficult to generate than iPSCs, the ability of somatic cell nuclear transfer to replace aged or diseased mitochondria and the closer epigenomic and transcriptomic similarity between NT-ESCs and bona fide ESCs may make NT-ESCs superior for future applications in regenerative medicine. Stem Cells 2017;35:26-34.

  20. Amino acid chloramine damage to proliferating cell nuclear antigen in mammalian cells.

    PubMed

    Salama, Samir A; Snapka, Robert M

    2012-01-01

    Amino acid chloramines (AACLs) are reactive secondary products of activated neutrophils. To understand AACL damage in cell nuclei, we exploited proliferating cell nuclear antigen (PCNA) as a nuclear protein damage reporter, using western blotting and mass spectrometry. Chloramines of proline, arginine, and glycine caused significant damage to PCNA in cells. Chloramines of taurine and histidine caused slight damage to PCNA in cells. Other AACLs caused no PCNA damage in intact cells. Evidence supports a sulfonamide, sulfinamide, or sulfenamide crosslinking mechanism involving cysteine 148 at the PCNA subunit interface, methionine sulfoxide formation as the basis of electrophoretic mobility shifting, and tyrosine and/or methionine residues as the likely targets of AACL damage to the PCNA antibody epitope. An interstitial fluid model experiment showed that physiological amino acids can mediate HOCl damage to PCNA in the presence of proteins that would otherwise completely quench the HOCl. PCNA is a sensitive biomarker of AACL damage in cell nuclei. Arginine chloramine and proline chloramine, or reactive species derived from them, were shown to enter cells and damage PCNA. Amino acids were shown to have at least two different mechanisms for suppressing PCNA damage in cells by their corresponding AACLs. Cysteine 148 was shown to be essential for PCNA subunit crosslinking by AACLs, and a crosslinking mechanism was proposed.

  1. Fission yeast Lem2 and Man1 perform fundamental functions of the animal cell nuclear lamina.

    PubMed

    Gonzalez, Yanira; Saito, Akira; Sazer, Shelley

    2012-01-01

    In animal cells the nuclear lamina, which consists of lamins and lamin-associated proteins, serves several functions: it provides a structural scaffold for the nuclear envelope and tethers proteins and heterochromatin to the nuclear periphery. In yeast, proteins and large heterochromatic domains including telomeres are also peripherally localized, but there is no evidence that yeast have lamins or a fibrous nuclear envelope scaffold. Nonetheless, we found that the Lem2 and Man1 proteins of the fission yeast Schizosaccharomyces pombe, evolutionarily distant relatives of the Lap2/Emerin/Man1 (LEM) sub-family of animal cell lamin-associated proteins, perform fundamental functions of the animal cell lamina. These integral inner nuclear membrane localized proteins, with nuclear localized DNA binding Helix-Extension-Helix (HEH) domains, impact nuclear envelope structure and integrity, are essential for the enrichment of telomeres at the nuclear periphery and by means of their HEH domains anchor chromatin, most likely transcriptionally repressed heterochromatin, to the nuclear periphery. These data indicate that the core functions of the nuclear lamina are conserved between fungi and animal cells and can be performed in fission yeast, without lamins or other intermediate filament proteins.

  2. Fission yeast Lem2 and Man1 perform fundamental functions of the animal cell nuclear lamina

    PubMed Central

    Gonzalez, Yanira; Saito, Akira; Sazer, Shelley

    2012-01-01

    In animal cells the nuclear lamina, which consists of lamins and lamin-associated proteins, serves several functions: it provides a structural scaffold for the nuclear envelope and tethers proteins and heterochromatin to the nuclear periphery. In yeast, proteins and large heterochromatic domains including telomeres are also peripherally localized, but there is no evidence that yeast have lamins or a fibrous nuclear envelope scaffold. Nonetheless, we found that the Lem2 and Man1 proteins of the fission yeast Schizosaccharomyces pombe, evolutionarily distant relatives of the Lap2/Emerin/Man1 (LEM) sub-family of animal cell lamin-associated proteins, perform fundamental functions of the animal cell lamina. These integral inner nuclear membrane localized proteins, with nuclear localized DNA binding Helix-Extension-Helix (HEH) domains, impact nuclear envelope structure and integrity, are essential for the enrichment of telomeres at the nuclear periphery and by means of their HEH domains anchor chromatin, most likely transcriptionally repressed heterochromatin, to the nuclear periphery. These data indicate that the core functions of the nuclear lamina are conserved between fungi and animal cells and can be performed in fission yeast, without lamins or other intermediate filament proteins. PMID:22540024

  3. Hot gas cleanup for molten carbonate fuel cells. A zinc oxide reactor model, Final report

    SciTech Connect

    Steinfeld, G.

    1980-09-16

    Utilization of coal gasifiers to power MCFC requires a cleanup system to remove sulfur and particulates. Of the two near term options available for desulfurization of gasifier effluent, namely low temperature cleanup utilizing absorber/stripper technology, and hot gas cleanup utilizing metal oxides, there is a clear advantage to using hot gas cleanup. Since the MCFC will operate at 1200/sup 0/F, and the gasifier effluent could be between 1200 to 1900/sup 0/F, a hot gas cleanup system will require little or no change in process gas temperature, thereby contributing to a high overall system efficiency. A hot gas cleanup system will consist of FeO for bulk H/sub 2/S removal and ZnO for reduction of H/sub 2/S to sub ppM levels. Hot gas cleanup systems at present are not available commercially, and therefore it is the objective of this project to model the components of the system in order to help bring this technology closer to commercialization, by providing simulated operating characteristics to aid in system design, and system simulations of gasifier/MCFC systems. The modeling of the ZnO reactor is presented.

  4. Nuclear orphan receptor TLX affects gene expression, proliferation and cell apoptosis in beta cells.

    PubMed

    Shi, Xiaoli; Xiong, Xiaokan; Dai, Zhe; Deng, Haohua; Sun, Li; Hu, Xuemei; Zhou, Feng; Xu, Yancheng

    Nuclear orphan receptor TLX is an essential regulator of the growth of neural stem cells. However, its exact function in pancreatic islet cells is still unknown. In the present study, gene expression profiling analysis revealed that overexpression of TLX in beta cell line MIN6 causes suppression of 176 genes and upregulation of 49 genes, including a cadre of cell cycle, cell proliferation and cell death control genes, such as Btg2, Ddit3 and Gadd45a. We next examined the effects of TLX overexpression on proliferation, apoptosis and insulin secretion in MIN6 cells. Proliferation analysis using EdU assay showed that overexpression of TLX increased percentage of EdU-positive cells. Cell cycle and apoptosis analysis revealed that overexpression of TLX in MIN6 cells resulted in higher percentage of cells exiting G1 into S-phase, and a 58.8% decrease of cell apoptosis induced by 0.5 mM palmitate. Moreover, TLX overexpression did not cause impairment of insulin secretion. Together, we conclude that TLX is among factors capable of controlling beta cell proliferation and survival, which may serve as a target for the development of novel therapies for diabetes.

  5. Research resource: Comparative nuclear receptor atlas: basal and activated peritoneal B-1 and B-2 cells.

    PubMed

    Diehl, Cody J; Barish, Grant D; Downes, Michael; Chou, Meng-Yun; Heinz, Sven; Glass, Christopher K; Evans, Ronald M; Witztum, Joseph L

    2011-03-01

    Naïve murine B cells are typically divided into three subsets based on functional and phenotypic characteristics: innate-like B-1 and marginal zone B cells vs. adaptive B-2 cells, also known as follicular or conventional B cells. B-1 cells, the innate-immune-like component of the B cell lineage are the primary source of natural antibodies and have been shown to modulate autoimmune diseases, human B-cell leukemias, and inflammatory disorders such as atherosclerosis. On the other hand, B-2 cells are the principal mediators of the adaptive humoral immune response and represent an important pharmacological target for various conditions including rheumatoid arthritis, lupus erythematosus, and lymphomas. Using the resources of the Nuclear Receptor Signaling Atlas program, we used quantitative real-time PCR to assess the complement of the 49 murine nuclear receptor superfamily expressed in quiescent and toll-like receptor (TLR)-stimulated peritoneal B-1 and B-2 cells. We report the expression of 24 nuclear receptors in basal B-1 cells and 25 nuclear receptors in basal B-2 cells, with, in some cases, dramatic changes in response to TLR 4 or TLR 2/1 stimulation. Comparative nuclear receptor profiling between B-1 and peritoneal B-2 cells reveals a highly concordant expression pattern, albeit at quantitatively dissimilar levels. We also found that splenic B cells express 23 nuclear receptors. This catalog of nuclear receptor expression in B-1 and B-2 cells provides data to be used to better understand the specific roles of nuclear receptors in B cell function, chronic inflammation, and autoimmune disease.

  6. Research Resource: Comparative Nuclear Receptor Atlas: Basal and Activated Peritoneal B-1 and B-2 Cells

    PubMed Central

    Diehl, Cody J.; Barish, Grant D.; Downes, Michael; Chou, Meng-Yun; Heinz, Sven; Glass, Christopher K.; Evans, Ronald M.

    2011-01-01

    Naïve murine B cells are typically divided into three subsets based on functional and phenotypic characteristics: innate-like B-1 and marginal zone B cells vs. adaptive B-2 cells, also known as follicular or conventional B cells. B-1 cells, the innate-immune-like component of the B cell lineage are the primary source of natural antibodies and have been shown to modulate autoimmune diseases, human B-cell leukemias, and inflammatory disorders such as atherosclerosis. On the other hand, B-2 cells are the principal mediators of the adaptive humoral immune response and represent an important pharmacological target for various conditions including rheumatoid arthritis, lupus erythematosus, and lymphomas. Using the resources of the Nuclear Receptor Signaling Atlas program, we used quantitative real-time PCR to assess the complement of the 49 murine nuclear receptor superfamily expressed in quiescent and toll-like receptor (TLR)-stimulated peritoneal B-1 and B-2 cells. We report the expression of 24 nuclear receptors in basal B-1 cells and 25 nuclear receptors in basal B-2 cells, with, in some cases, dramatic changes in response to TLR 4 or TLR 2/1 stimulation. Comparative nuclear receptor profiling between B-1 and peritoneal B-2 cells reveals a highly concordant expression pattern, albeit at quantitatively dissimilar levels. We also found that splenic B cells express 23 nuclear receptors. This catalog of nuclear receptor expression in B-1 and B-2 cells provides data to be used to better understand the specific roles of nuclear receptors in B cell function, chronic inflammation, and autoimmune disease. PMID:21273443

  7. Nuclear Survivin Expression in Mantle Cell Lymphoma Is Associated with Cell Proliferation and Survival

    PubMed Central

    Martinez, Antonio; Bellosillo, Beatriz; Bosch, Francesc; Ferrer, Ana; Marcé, Silvia; Villamor, Neus; Ott, German; Montserrat, Emili; Campo, Elias; Colomer, Dolors

    2004-01-01

    Survivin is a member of the inhibitor of apoptosis protein family that is expressed in G2/M phase. Survivin is overexpressed and associated with parameters of poor prognosis in different human tumors. The role of survivin in the pathogenesis of mantle cell lymphoma (MCL) was examined in a series of typical and blastoid tumors. Survivin was detected as a nuclear pattern in a variable number of tumor cells. Mitotic figures were always positive with a strong delineation of the chromosomes. Western blot analysis confirmed the presence of survivin only in nuclear fractions. Protein expression detected by immunohistochemistry correlated with mRNA levels analyzed by quantitative real-time reverse transcription-polymerase chain reaction (P < 0.0001). Survivin expression levels were higher in blastoid MCL variants (P < 0.0001) and were associated with the proliferative activity (P = 0.001), but not with the ploidy status of the tumors. The number of apoptotic cells was independent of survivin or Ki-67 expression. Overall survival was significantly shorter in patients with high survivin expression. However, in a multivariate analysis, proliferative index was a better predictor of survival than survivin score. These findings indicate that survivin is commonly expressed in MCL with a nuclear and mitotic pattern. The expression levels are strongly associated with the proliferative activity of the tumors and the survival of the patients, suggesting a potential role in cell cycle regulation and tumor progression. PMID:14742256

  8. Differential nuclear remodeling of mammalian somatic cells by Xenopus laevis oocyte and egg cytoplasm

    SciTech Connect

    Alberio, Ramiro; Johnson, Andrew D.; Stick, Reimer; Campbell, Keith H.S. . E-mail: keith.campbell@nottingham.ac.uk

    2005-07-01

    The mechanisms governing nuclear reprogramming have not been fully elucidated yet; however, recent studies show a universally conserved ability of both oocyte and egg components to reprogram gene expression in somatic cells. The activation of genes associated with pluripotency by oocyte/egg components may require the remodeling of nuclear structures, such that they can acquire the features of early embryos and pluripotent cells. Here, we report on the remodeling of the nuclear lamina of mammalian cells by Xenopus oocyte and egg extracts. Lamin A/C is removed from somatic cells incubated in oocyte and egg extracts in an active process that requires permeable nuclear pores. Removal of lamin A/C is specific, since B-type lamins are not changed, and it is not dependent on the incorporation Xenopus egg specific lamin III. Moreover, transcriptional activity is differentially regulated in somatic cells incubated in the extracts. Pol I and II transcriptions are maintained in cells in oocyte extracts; however, both activities are abolished in egg extracts. Our study shows that components of oocyte and egg extracts can modify the nuclear lamina of somatic cells and that this nuclear remodeling induces a structural change in the nucleus which may have implications for transcriptional activity. These experiments suggest that modifications in the nuclear lamina structure by the removal of somatic proteins and the incorporation of oocyte/egg components may contribute to the reprogramming of somatic cell nuclei and may define a characteristic configuration of pluripotent cells.

  9. The nuclear membrane and mechanotransduction: impaired nuclear mechanics and mechanotransduction in lamin A/C deficient cells.

    PubMed

    Lammerding, Jan; Lee, Richard T

    2005-01-01

    Mutations in the lamin A/C gene cause a variety of human diseases including Emery-Dreifuss muscular dystrophy, dilated cardiomyopathy and Hutchinson-Gilford progeria syndrome. The tissue specific effects of lamin mutations are unclear, in part because the function of lamin A/C is incompletely defined, but the many muscle specific phenotypes suggest that defective lamin A/C could increase cellular mechanical sensitivity. Lamin A/C deficient fibroblasts were subjected to mechanical strain to measure nuclear mechanical properties and strain-induced signalling. We found that lamin A/C deficient fibroblasts are characterized by impaired nuclear mechanics and mechanotransduction, reflected by increased nuclear deformations, increased nuclear fragility, attenuated expression of mechanosensitive genes, and impaired transcriptional activation, leading to impaired viability of mechanically strained cells. Lamins and other nuclear envelope proteins can thus affect several levels of the mechanotransduction cascade, altering nuclear and cytoskeletal mechanics as well as playing an important role in mechanically activated gene regulation. Individual mutations in the lamin A/C gene could potentially selectively interfere with any of these functions, explaining the tissue-specific effects observed in the laminopathies.

  10. Nuclear reprogramming to produce cloned mice and embryonic stem cells from somatic cells.

    PubMed

    Wakayama, Sayaka; Cummins, James M; Wakayama, Teruhiko

    2008-04-01

    Cloning methods in mice are now well described and are becoming routine. However, the frequency at which cloned mice are produced remains below 5%, irrespective of the nucleus donor species or cell type. Only a few laboratories have made clones from adult mouse somatic cells and most strains have never produced cloned mice. On the other hand, nuclear transfer can be used to generate human embryonic stem (ntES) cell lines from a patient's own somatic cells. It has been shown that such cells can be generated relatively easily from a variety of mouse genotypes and cell types of both sexes, even though it may be more difficult to generate clones directly. This technique could be used in regenerative medicine and, in theory, in infertility clinics to treat completely infertile individuals. However, these results suggest that the reprogramming integrity of each cloned embryo differs: some cloned embryos can be converted to ntES cells, but these embryos cannot achieve full term development. This review outlines the nature of genomic reprogramming potential and its application, and suggests new approaches to avoid the ethical problems of creating embryos by nuclear transfer.

  11. Detection of Endogenous Nuclear Proteins in Plant Cells: Localizing Nuclear Matrix Constituent Proteins (NMCPs), the Plant Analogs of Lamins.

    PubMed

    Ciska, Malgorzata; de la Espina, Susana Moreno Díaz

    2017-01-01

    At present, two complementary approaches are used for in situ protein visualization in plant nuclei. Imaging of transformed fluorescent proteins is the election tool for the analysis of protein movement and interaction. However, this methodology presents several drawbacks for the identification/localization of endogenous nuclear factors, such as over-expression or mislocalization of transformed proteins. In contrast, immunocytochemistry with specific antibodies represents a powerful tool for the localization of endogenous nuclear proteins at their "native" nuclear sub-compartments. In plant cells, the cell wall hampers antibody accessibility during immunocytochemical analysis thereby reducing the effectivity of the technique, particularly in the case of lowly expressed proteins. To overcome this problem in nuclear protein immunodetection, we developed a method based on the in vitro incubation of isolated nuclei with specific antibodies followed by imaging by confocal fluorescence or electron microscopy. Here we describe the application of this methodology to the localization of Nuclear Matrix Constituent Proteins (NMCP), the plant analogs of lamins, of the monocot Allium cepa, using antibodies raised against highly conserved regions of the proteins.

  12. Physical and functional interaction of human nuclear uracil-DNA glycosylase with proliferating cell nuclear antigen☆

    PubMed Central

    Ko, Rinkei; Bennett, Samuel E.

    2011-01-01

    Uracil residues arise in DNA by the misincorporation of dUMP in place of dTMP during DNA replication or by the deamination of cytosine in DNA. Uracil-DNA glycosylase initiates DNA base excision repair of uracil residues by catalyzing the hydrolysis of the N-glycosylic bond linking the uracil base to deoxyribose. In human cells, the nuclear form of uracil-DNA glycosylase (UNG2) contains a conserved PCNA-binding motif located at the N-terminus that has been implicated experimentally in binding PCNA. Here we use purified preparations of UNG2 and PCNA to demonstrate that UNG2 physically associates with PCNA. UNG2 co-eluted with PCNA during size exclusion chromatography and bound to a PCNA affinity column. Association of UNG2 with PCNA was abolished by the addition of 100 mM NaCl, and significantly decreased in the presence of 10 mM MgCl2. The functional significance of the UNG2·PCNA association was demonstrated by UNG2 activity assays. Addition of PCNA (30–810 pmol) to standard uracil-DNA glycosylase reactions containing linear [uracil-3H]DNA stimulated UNG2 catalytic activity up to 2.6-fold. UNG2 activity was also stimulated by 7.5 mM MgCl2. The stimulatory effect of PCNA was increased by the addition of MgCl2; however, the dependence on PCNA concentration was the same, indicating that the effects of MgCl2 and PCNA on UNG2 activity occurred by independent mechanisms. Loading of PCNA onto the DNA substrate was required for stimulation, as the activity of UNG2 on circular DNA substrates was not affected by the addition of PCNA. Addition of replication factor C and ATP to reactions containing 90 pmol of PCNA resulted in two-fold stimulation of UNG2 activity on circular DNA. PMID:16216562

  13. Slip-cast and hot-solution infiltrated porous yttria stabilized zirconia (YSZ) supported tubular fuel cells

    NASA Astrophysics Data System (ADS)

    Hanifi, Amir Reza; Paulson, Scott; Torabi, Alireza; Shinbine, Alyssa; Tucker, Michael C.; Birss, Viola; Etsell, Thomas H.; Sarkar, Partha

    2014-11-01

    Hot solution infiltration was investigated as a flexible and rapid method to incorporate anode and cathode components into fully sintered, porous ceramic tubular templates for use as solid oxide fuel cells (SOFC). Composed of either a porous 8 mol% yttria-stabilized zirconia (YSZ) or 5 wt% NiO-YSZ support structure, a thin Ni-YSZ anode functional layer and an outer ca. 10 μm dense YSZ electrolyte, closed end tubes were first hot solution (ca. 100 °C) infiltrated on the inside with NiO-SDC (Sm0.2Ce0.8O1.9) to serve as the anode. Cathodes were either LSM (nominally La0.8Sr0.2MnO3+δ) infiltrated into a thin porous YSZ layer on the outer electrolyte surface, or an LSCF-GDC composite (Gd0.1Ce0.9O1.95-La0.6Sr0.4Co0.2Fe0.8O3-δ) on a thin GDC buffer layer. Although hot solution infiltration of the Ni, Ce and Sm salts into the anode support structure did not result in complete penetration (with the Ni contents in the tube wall ranging between 4 and 10 vol.%), well-sealed full cells produced power densities as high as 275, 196 and 153 mW cm-2 at 800, 750 and 700 °C, respectively. Hot solution infiltration of active SOFC electrode materials is thus shown to be a very flexible approach for the evaluation of their performance.

  14. Karyopherin Alpha 1 Regulates Satellite Cell Proliferation and Survival by Modulating Nuclear Import.

    PubMed

    Choo, Hyo-Jung; Cutler, Alicia; Pavlath, Grace K

    2016-07-19

    Satellite cells are stem cells with an essential role in skeletal muscle repair. Precise regulation of gene expression is critical for proper satellite cell quiescence, proliferation, differentiation and self-renewal. Nuclear proteins required for gene expression are dependent on the nucleocytoplasmic transport machinery to access to nucleus, however little is known about regulation of nuclear transport in satellite cells. The best characterized nuclear import pathway is classical nuclear import which depends on a classical nuclear localization signal (cNLS) in a cargo protein and the heterodimeric import receptors, karyopherin alpha (KPNA) and beta (KPNB). Multiple KPNA1 paralogs exist and can differ in importing specific cNLS proteins required for cell differentiation and function. We show that transcripts for six Kpna paralogs underwent distinct changes in mouse satellite cells during muscle regeneration accompanied by changes in cNLS proteins in nuclei. Depletion of KPNA1, the most dramatically altered KPNA, caused satellite cells in uninjured muscle to prematurely activate, proliferate and undergo apoptosis leading to satellite cell exhaustion with age. Increased proliferation of satellite cells led to enhanced muscle regeneration at early stages of regeneration. In addition, we observed impaired nuclear localization of two key KPNA1 cargo proteins: p27, a cyclin-dependent kinase inhibitor associated with cell cycle control and lymphoid enhancer factor 1, a critical cotranscription factor for β-catenin. These results indicate that regulated nuclear import of proteins by KPNA1 is critical for satellite cell proliferation and survival and establish classical nuclear import as a novel regulatory mechanism for controlling satellite cell fate. Stem Cells 2016.

  15. Coordination between donor cell type and cell cycle stage improves nuclear cloning efficiency in cattle.

    PubMed

    Wells, D N; Laible, G; Tucker, F C; Miller, A L; Oliver, J E; Xiang, T; Forsyth, J T; Berg, M C; Cockrem, K; L'Huillier, P J; Tervit, H R; Oback, B

    2003-01-01

    Several studies have shown that both quiescent and proliferating somatic donor cells can be fully reprogrammed after nuclear transfer (NT) and result in viable offspring. So far, however, no comparative study has conclusively demonstrated the relative importance of donor cell cycle stage on nuclear cloning efficiency. Here, we compare two different types of bovine fetal fibroblasts (BFFs) that were synchronized in G(0), G(1), and different phases within G(1). We show that for non-transgenic (non-TG) fibroblasts, serum starvation into G(0) results in a significantly higher percentage of viable calves at term than synchronization in early G(1) or late G(1). For transgenic fibroblasts, however, cells selected in G(1) show significantly higher development to calves at term and higher post-natal survival to weaning than cells in G(0). This suggests that it may be necessary to coordinate donor cell type and cell cycle stage to maximize overall cloning efficiency. Copyright 2002 Elsevier Science Inc.

  16. Dnmt3l-knockout donor cells improve somatic cell nuclear transfer reprogramming efficiency.

    PubMed

    Liao, Hung-Fu; Mo, Chu-Fan; Wu, Shinn-Chih; Cheng, Dai-Han; Yu, Chih-Yun; Chang, Kai-Wei; Kao, Tzu-Hao; Lu, Chia-Wei; Pinskaya, Marina; Morillon, Antonin; Lin, Shih-Shun; Cheng, Winston T K; Bourc'his, Déborah; Bestor, Timothy; Sung, Li-Ying; Lin, Shau-Ping

    2015-10-01

    Nuclear transfer (NT) is a technique used to investigate the development and reprogramming potential of a single cell. DNA methyltransferase-3-like, which has been characterized as a repressive transcriptional regulator, is expressed in naturally fertilized egg and morula/blastocyst at pre-implantation stages. In this study, we demonstrate that the use of Dnmt3l-knockout (Dnmt3l-KO) donor cells in combination with Trichostatin A treatment improved the developmental efficiency and quality of the cloned embryos. Compared with the WT group, Dnmt3l-KO donor cell-derived cloned embryos exhibited increased cell numbers as well as restricted OCT4 expression in the inner cell mass (ICM) and silencing of transposable elements at the blastocyst stage. In addition, our results indicate that zygotic Dnmt3l is dispensable for cloned embryo development at pre-implantation stages. In Dnmt3l-KO mouse embryonic fibroblasts, we observed reduced nuclear localization of HDAC1, increased levels of the active histone mark H3K27ac and decreased accumulation of the repressive histone marks H3K27me3 and H3K9me3, suggesting that Dnmt3l-KO donor cells may offer a more permissive epigenetic state that is beneficial for NT reprogramming.

  17. Nuclear-mitochondrial incompatibility in interorder rhesus monkey-cow embryos derived from somatic cell nuclear transfer.

    PubMed

    Kwon, Daekee; Koo, Ok-Jae; Kim, Min-Jung; Jang, Goo; Lee, Byeong Chun

    2016-10-01

    Monkey interorder somatic cell nuclear transfer (iSCNT) using enucleated cow oocytes yielded poor blastocysts development and contradictory results among research groups. Determining the reason for this low blastocyst development is a prerequisite for optimizing iSCNT in rhesus monkeys. The aim of this study was to elucidate nuclear-mitochondrial incompatibility of rhesus monkey-cow iSCNT embryos and its relationship to low blastocyst development. Cytochrome b is a protein of complex III of the electron transport chain (ETC). According to meta-analysis of amino acid sequences, the homology of cytochrome b is 75 % between rhesus monkeys and cattle. To maintain the function of ETC after iSCNT, 4n iSCNT embryos were produced by fusion of non-enucleated cow oocytes and rhesus monkey somatic cells. The blastocyst development rate of 4n iSCNT embryos was higher than that of 2n embryos (P < 0.01). Formation of reactive oxygen species (ROS) is an indirect indicator of ETC activity of cells. The ROS levels of 4n iSCNT embryos was higher than that of 2n embryos (P < 0.01). Collectively, rhesus monkey iSCNT embryos reconstructed with cow oocytes have nuclear-mitochondrial incompatibility due to fundamental species differences between rhesus monkeys and cattle. Nuclear-mitochondrial incompatibility seems to correlate with low ETC activity and extremely low blastocyst development of rhesus monkey-cow iSCNT embryos.

  18. SUMOylation regulates the nuclear mobility of CREB binding protein and its association with nuclear bodies in live cells

    SciTech Connect

    Ryan, Colm M.; Kindle, Karin B.; Collins, Hilary M.; Heery, David M.

    2010-01-01

    The lysine acetyltransferase CREB binding protein (CBP) is required for chromatin modification and transcription at many gene promoters. In fixed cells, a large proportion of CBP colocalises to PML or nuclear bodies. Using live cell imaging, we show here that YFP-tagged CBP expressed in HEK293 cells undergoes gradual accumulation in nuclear bodies, some of which are mobile and migrate towards the nuclear envelope. Deletion of a short lysine-rich domain that contains the major SUMO acceptor sites of CBP abrogated its ability to be SUMO modified, and prevented its association with endogenous SUMO-1/PML speckles in vivo. This SUMO-defective CBP showed enhanced ability to co-activate AML1-mediated transcription. Deletion mapping revealed that the SUMO-modified region was not sufficient for targeting CBP to PML bodies, as C-terminally truncated mutants containing this domain showed a strong reduction in accumulation at PML bodies. Fluorescence recovery after photo-bleaching (FRAP) experiments revealed that YFP-CBP{Delta}998-1087 had a retarded recovery time in the nucleus, as compared to YFP-CBP. These results indicate that SUMOylation regulates CBP function by influencing its shuttling between nuclear bodies and chromatin microenvironments.

  19. Inhibition of thromboxane synthase induces lung cancer cell death via increasing the nuclear p27

    SciTech Connect

    Leung, Kin Chung; Hsin, Michael K.Y.; Chan, Joey S.Y.; Yip, Johnson H.Y.; Li, Mingyue; Leung, Billy C.S.; Mok, Tony S.K.; Warner, Timothy D.; Underwood, Malcolm J.; Chen, George G.

    2009-10-15

    The role of thromboxane in lung carcinogenesis is not clearly known, though thromboxane B2 (TXB{sub 2}) level is increased and antagonists of thromboxane receptors or TXA2 can induce apoptosis of lung cancer cells. p27, an atypical tumor suppressor, is normally sequestered in the nucleus. The increased nuclear p27 may result in apoptosis of tumor cells. We hypothesize that the inhibition of thromboxane synthase (TXS) induces the death of lung cancer cells and that such inhibition is associated with the nuclear p27 level. Our experiment showed that the inhibition of TXS significantly induced the death or apoptosis in lung cancer cells. The activity of TXS was increased in lung cancer. The nuclear p27 was remarkably reduced in lung cancer tissues. The inhibition of TXS caused the cell death and apoptosis of lung cancer cells, likely via the elevation of the nuclear p27 since the TXS inhibition promoted the nuclear p27 level and the inhibition of p27 by its siRNA recovered the cell death induced by TXS inhibition. Collectively, lung cancer cells produce high levels of TXB{sub 2} but their nuclear p27 is markedly reduced. The inhibition of TXS results in the p27-related induction of cell death in lung cancer cells.

  20. Subclassification of prostate cancer circulating tumor cells by nuclear size reveals very small nuclear circulating tumor cells in patients with visceral metastases.

    PubMed

    Chen, Jie-Fu; Ho, Hao; Lichterman, Jake; Lu, Yi-Tsung; Zhang, Yang; Garcia, Mitch A; Chen, Shang-Fu; Liang, An-Jou; Hodara, Elisabeth; Zhau, Haiyen E; Hou, Shuang; Ahmed, Rafi S; Luthringer, Daniel J; Huang, Jiaoti; Li, Ker-Chau; Chung, Leland W K; Ke, Zunfu; Tseng, Hsian-Rong; Posadas, Edwin M

    2015-09-15

    Although enumeration of circulating tumor cells (CTCs) has shown some clinical value, the pool of CTCs contains a mixture of cells that contains additional information that can be extracted. The authors subclassified CTCs by shape features focusing on nuclear size and related this with clinical information. A total of 148 blood samples were obtained from 57 patients with prostate cancer across the spectrum of metastatic states: no metastasis, nonvisceral metastasis, and visceral metastasis. CTCs captured and enumerated on NanoVelcro Chips (CytoLumina, Los Angeles, Calif) were subjected to pathologic review including nuclear size. The distribution of nuclear size was analyzed using a Gaussian mixture model. Correlations were made between CTC subpopulations and metastatic status. Statistical modeling of nuclear size distribution revealed 3 distinct subpopulations: large nuclear CTCs, small nuclear CTCs, and very small nuclear CTCs (vsnCTCs). Small nuclear CTCs and vsnCTC identified those patients with metastatic disease. However, vsnCTC counts alone were found to be elevated in patients with visceral metastases when compared with those without (0.36 ± 0.69 vs 1.95 ± 3.77 cells/mL blood; P<.001). Serial enumeration studies suggested the emergence of vsnCTCs occurred before the detection of visceral metastases. There are morphologic subsets of CTCs that can be identified by fundamental pathologic approaches, such as nuclear size measurement. The results of this observational study strongly suggest that CTCs contain relevant information regarding disease status. In particular, the detection of vsnCTCs was found to be correlated with the presence of visceral metastases and should be formally explored as a putative blood-borne biomarker to identify patients at risk of developing this clinical evolution of prostate cancer. © 2015 American Cancer Society.

  1. Prediction of radiosensitivity in human bladder cell lines using nuclear chromatin phenotype.

    PubMed

    Rajab, Nor F; McKenna, Declan J; Diamond, Jim; Williamson, Kate; Hamilton, Peter W; McKelvey-Martin, Valerie J

    2006-10-01

    Nuclear texture analysis measures phenotypic changes in chromatin distribution within a cell nucleus, while the alkaline Comet assay is a sensitive method for measuring the extent of DNA breakage in individual cells. The authors aim to use both methods to provide information about the sensitivity of cells to ionizing radiation. The alkaline Comet assay was performed on six human bladder carcinoma cell lines and one human urothelial cell line exposed to gamma-radiation doses from 0 to 10 Gy. Nuclear chromatin texture analysis of 40 features was then performed in the same cell lines exposed to 0, 2, and 6 Gy to explore if nuclear phenotype was related to radiation sensitivity. Comet assay results demonstrated that the cell lines exhibited different levels of radiosensitivity and could be divided into a radiosensitive and a radioresistant group at >6 Gy. Using stepwise discriminant analysis, a subset of important nuclear texture features that best discriminated between sensitive and resistant cell lines were identified A classification function, defined using these features, correctly classified 81.75% of all cells into their radiosensitive or radioresistant groups based on their pretreatment chromatin phenotype. Posttreatment chromatin changes also varied between cell lines, with sensitive cell lines showing a relaxed chromatin conformation following radiation, whereas resistant cell lines exhibited chromatin condensation. The authors conclude that the alkaline Comet assay and nuclear texture methodologies may prove to be valuable aids in predicting the response of tumor cells to radiotherapy.

  2. Nuclear receptor atlas of female mouse liver parenchymal, endothelial, and Kupffer cells.

    PubMed

    Li, Zhaosha; Kruijt, J Kar; van der Sluis, Ronald J; Van Berkel, Theo J C; Hoekstra, Menno

    2013-04-01

    The liver consists of different cell types that together synchronize crucial roles in liver homeostasis. Since nuclear receptors constitute an important class of drug targets that are involved in a wide variety of physiological processes, we have composed the hepatic cell type-specific expression profile of nuclear receptors to uncover the pharmacological potential of liver-enriched nuclear receptors. Parenchymal liver cells (hepatocytes) and liver endothelial and Kupffer cells were isolated from virgin female C57BL/6 wild-type mice using collagenase perfusion and counterflow centrifugal elutriation. The hepatic expression pattern of 49 nuclear receptors was generated by real-time quantitative PCR using the NUclear Receptor Signaling Atlas (NURSA) program resources. Thirty-six nuclear receptors were expressed in total liver. FXR-α, EAR2, LXR-α, HNF4-α, and CAR were the most abundantly expressed nuclear receptors in liver parenchymal cells. In contrast, NUR77, COUP-TFII, LXR-α/β, FXR-α, and EAR2 were the most highly expressed nuclear receptors in endothelial and Kupffer cells. Interestingly, members of orphan receptor COUP-TF family showed a distinct expression pattern. EAR2 was highly and exclusively expressed in parenchymal cells, while COUP-TFII was moderately and exclusively expressed in endothelial and Kupffer cells. Of interest, the orphan receptor TR4 showed a similar expression pattern as the established lipid sensor PPAR-γ. In conclusion, our study provides the most complete quantitative assessment of the nuclear receptor distribution in liver reported to date. Our gene expression catalog suggests that orphan nuclear receptors such as COUP-TFII, EAR2, and TR4 may be of significant importance as novel targets for pharmaceutical interventions in liver.

  3. Birth of Beagle dogs by somatic cell nuclear transfer.

    PubMed

    Hossein, Mohammad Shamim; Jeong, Yeon Woo; Park, Sun Woo; Kim, Joung Joo; Lee, Eugine; Ko, Kyeong Hee; Hyuk, Park; Hoon, Song Seung; Kim, Yeun Wook; Hyun, Sang Hwan; Shin, Taeyoung; Hwang, Woo Suk

    2009-09-01

    The present study was undertaken to evaluate two enucleation methods for somatic cell nuclear transfer (SCNT), and to standardize the optimum number of embryos for transfer to each recipient for canines. Oocytes retrieved from outbreed dogs were reconstructed with adult somatic cells from a male Beagle dog. A total of 134 or 267 oocytes were enucleated either by aspiration or squeezing method, fused with two DC pulses of 1.75 kV/cm for 15 micros electrical stimulation, chemically activated after 1h of fusion using 10 microM calcium ionophore for 4 min and cultured 4h in 1.9 mM 6-dimethylaminopurine. Finally, 103 or 214 embryos for aspiration or squeezing method were transferred to 6 or 11 naturally synchronized recipients, respectively. A total of 53, 317 and 342 embryos were transferred to 7, 17 and 12 recipients for the group of 4-10, 11-25 and 26-40 embryos, respectively. There was no difference between fusion rate (76.87% vs. 80.15%), full term pregnancy rate (16.66% vs. 27.27%) and percent of live puppies born (0.97% vs. 1.87%) for aspiration and squeezing method (P>0.05). Production efficiency of cloned dogs was significantly affected by the number of embryos transferred to each recipient. No pregnancy was established for the group of 4-10 embryos (n=7) and 26-40 embryos (n=12) while pregnancy was detected in 23.53% recipients received a group of 11-25 embryos (n=17). Among them, five (1.76%) live puppies were born (P<0.05). These data show an increase in the overall efficiency of SCNT in canine species.

  4. Hot-cell design considerations for interfacing eddy-current systems

    SciTech Connect

    Franklin, E.M.; Webb, J.P.; Larson, J.M.

    1985-01-01

    The Hot Fuel Examination Facility/North conducts remote eddy-current examination of irradiated fuel elements. Applications include cladding breach detection and irradiation-induced ferrite examination. The seccussful use of remote eddy-current techniques is achieved by applying basic test parameters and interfacing considerations. These include impedance matching, operating frequency, and feedthrough considerations.

  5. Hot Flashes

    MedlinePlus

    ... are due to menopause — the time when menstrual periods become irregular and eventually stop. In fact, hot flashes are the most common symptom of the menopausal transition. How often hot flashes occur varies among women ...

  6. Karyopherin alpha 1 regulates satellite cell proliferation and survival by modulating nuclear import

    PubMed Central

    Choo, Hyo-Jung; Cutler, Alicia; Rother, Franziska; Bader, Michael; Pavlath, Grace K.

    2016-01-01

    Satellite cells are stem cells with an essential role in skeletal muscle repair. Precise regulation of gene expression is critical for proper satellite cell quiescence, proliferation, differentiation and self -renewal. Nuclear proteins required for gene expression are dependent on the nucleocytoplasmic transport machinery to access to nucleus, however little is known about regulation of nuclear transport in satellite cells. The best characterized nuclear import pathway is classical nuclear import which depends on a classical nuclear localization signal (cNLS) in a cargo protein and the heterodimeric import receptors, karyopherin alpha (KPNA) and beta (KPNB). Multiple KPNA1 paralogs exist and can differ in importing specific cNLS proteins required for cell differentiation and function. We show that transcripts for six Kpna paralogs underwent distinct changes in mouse satellite cells during muscle regeneration accompanied by changes in cNLS proteins in nuclei. Depletion of KPNA1, the most dramatically altered KPNA, caused satellite cells in uninjured muscle to prematurely activate, proliferate and undergo apoptosis leading to satellite cell exhaustion with age. Increased proliferation of satellite cells led to enhanced muscle regeneration at early stages of regeneration. In addition, we observed impaired nuclear localization of two key KPNA1 cargo proteins: p27, a cyclin-dependent kinase inhibitor associated with cell cycle control and lymphoid enhancer factor 1, a critical co-transcription factor for β-catenin. These results indicate that regulated nuclear import of proteins by KPNA1 is critical for satellite cell proliferation and survival and establish classical nuclear import as a novel regulatory mechanism for controlling satellite cell fate. PMID:27434733

  7. Cell-free extract from porcine induced pluripotent stem cells can affect porcine somatic cell nuclear reprogramming.

    PubMed

    No, Jin-Gu; Choi, Mi-Kyung; Kwon, Dae-Jin; Yoo, Jae Gyu; Yang, Byoung-Chul; Park, Jin-Ki; Kim, Dong-Hoon

    2015-01-01

    Pretreatment of somatic cells with undifferentiated cell extracts, such as embryonic stem cells and mammalian oocytes, is an attractive alternative method for reprogramming control. The properties of induced pluripotent stem cells (iPSCs) are similar to those of embryonic stem cells; however, no studies have reported somatic cell nuclear reprogramming using iPSC extracts. Therefore, this study aimed to evaluate the effects of porcine iPSC extracts treatment on porcine ear fibroblasts and early development of porcine cloned embryos produced from porcine ear skin fibroblasts pretreated with the porcine iPSC extracts. The Chariot(TM) reagent system was used to deliver the iPSC extracts into cultured porcine ear skin fibroblasts. The iPSC extracts-treated cells (iPSC-treated cells) were cultured for 3 days and used for analyzing histone modification and somatic cell nuclear transfer. Compared to the results for nontreated cells, the trimethylation status of histone H3 lysine residue 9 (H3K9) in the iPSC-treated cells significantly decreased. The expression of Jmjd2b, the H3K9 trimethylation-specific demethylase gene, significantly increased in the iPSC-treated cells; conversely, the expression of the proapoptotic genes, Bax and p53, significantly decreased. When the iPSC-treated cells were transferred into enucleated porcine oocytes, no differences were observed in blastocyst development and total cell number in blastocysts compared with the results for control cells. However, H3K9 trimethylation of pronuclear-stage-cloned embryos significantly decreased in the iPSC-treated cells. Additionally, Bax and p53 gene expression in the blastocysts was significantly lower in iPSC-treated cells than in control cells. To our knowledge, this study is the first to show that an extracts of porcine iPSCs can affect histone modification and gene expression in porcine ear skin fibroblasts and cloned embryos.

  8. Nuclear survivin promoted by acetylation is associated with the aggressive phenotype of oral squamous cell carcinoma.

    PubMed

    Liu, Shuli; Shi, Lei; Yang, Xi; Ye, Dongxia; Wang, Tong; Dong, Cunshan; Guo, Wenzheng; Liao, Yueling; Song, Hongyong; Xu, Dongliang; Hu, Jingzhou; Zhang, Zhiyuan; Deng, Jiong

    2017-04-06

    Defects in apoptotic pathway contribute to development and progression of oral cancer. Survivin, a member of the inhibitors of apoptosis protein (IAP) family, is increased in many types of cancers. However, it is unclear whether increased survivin is associated with oral squamous cell carcinomas (OSCC), and what mechanisms may involve in. In this study, we examined survivin expression in OSCC compared to normal oral tissues via immunohistochemical staining. The results showed that, not only total survivin is increased in OSCCs, but also the subcellular location of survivin is changed in OSCCs compared to normal oral tissues. In most of normal oral tissues, survivin staining was either negative, or cytoplasmic positive/nuclear negative; whereas in most of OSCC tissues, survivin staining was nuclear positive. Statistic analysis indicates that nuclear survivin, rather than total or cytoplasmic one, correlates with tumor TNM stage and differentiation grade. Consistently, in vitro analysis showed that survivin is in cytoplasm in normal human oral kinotinocyte (HOK) cells; whereas it is in nucleus in OSCC HN6 cells. Importantly, treatment of HOK cells with HDAC inhibitor Trichostatin A (TSA) induces survivin acetylation and promotes its nuclear localization. Moreover, nuclear survivin in OSCC cells was acetylated at K129 in its C-terminal, suggesting that the acetylation is important for nuclear location of survivin. Our study demonstrates that it is nuclear survivin, rather than total or cytoplasmic one, associates with TNM stage and tumor grade of OSCC. Thus, we propose nuclear survivin as a prognostic marker for the progression of OSCC.

  9. Cell cycle synchronization of canine ear fibroblasts for somatic cell nuclear transfer.

    PubMed

    Koo, Ok Jae; Hossein, Mohammad Shamim; Hong, So Gun; Martinez-Conejero, Jose A; Lee, Byeong Chun

    2009-02-01

    Cycle synchronization of donor cells in the G0/G1 stage is a crucial step for successful somatic cell nuclear transfer. In the present report, we evaluated the effects of contact inhibition, serum starvation and the reagents - dimethyl sulphoxide (DMSO), roscovitine and cycloheximide (CHX) - on synchronization of canine fibroblasts at the G0/G1 stage. Ear fibroblast cells were collected from a beagle dog, placed into culture and used for analysis at passages three to eight. The population doubling time was 36.5 h. The proportion of G0/G1 cells was significantly increased by contact inhibition (77.1%) as compared with cycling cells (70.1%); however, extending the duration of culture did not induce further synchronization. After 24 h of serum starvation, cells were effectively synchronized at G0/G1 (77.1%). Although synchronization was further increased gradually after 24 h and even showed significant difference after 72 h (82.8%) of starvation, the proportion of dead cells also significantly increased after 24 h. The percentage of cells at the G0/G1 phase was increased (as compared with controls) after 72 h treatment with DMSO (76.1%) and after 48 h treatment with CHX (73.0%) or roscovitine (72.5%). However, the rate of cell death was increased after 24 and 72 h of treatment with DMSO and CHX, respectively. Thus, we recommend the use of roscovitine for cell cycle synchronization of canine ear fibroblasts as a preparatory step for SCNT.

  10. Hot conditioning equipment conceptual design report

    SciTech Connect

    Bradshaw, F.W., Westinghouse Hanford

    1996-08-06

    This report documents the conceptual design of the Hot Conditioning System Equipment. The Hot conditioning System will consist of two separate designs: the Hot Conditioning System Equipment; and the Hot Conditioning System Annex. The Hot Conditioning System Equipment Design includes the equipment such as ovens, vacuum pumps, inert gas delivery systems, etc.necessary to condition spent nuclear fuel currently in storage in the K Basins of the Hanford Site. The Hot Conditioning System Annex consists of the facility of house the Hot Conditioning System. The Hot Conditioning System will be housed in an annex to the Canister Storage Building. The Hot Conditioning System will consist of pits in the floor which contain ovens in which the spent nuclear will be conditioned prior to interim storage.

  11. Nuclear Targeted Silver Nanospheres Perturb the Cancer Cell Cycle Differently than those of Nanogold

    PubMed Central

    Austin, Lauren A.; Kang, Bin; Yen, Chun-Wan; El-Sayed, Mostafa A.

    2016-01-01

    Plasmonic nanoparticle research has become increasingly active due to their potential uses in biomedical applications. However, little is known about the intracellular effects these nanoparticles have on mammalian cells. The aim of this work is to investigate whether silver nanoparticles (AgNPs) conjugated with nuclear and cytoplasmic targeting peptides exhibit the same intracellular effects on cancer cells as peptide-conjugated gold nanoparticles (AuNPs). Nuclear and cytoplasmic targeting spherical AgNPs with a diameter of 35 nm were incubated in a cancer (HSC-3) and healthy (HaCat) cell line. By utilizing flow cytometry, confocal microscopy, and real-time dark field imaging, we were able to analyze how targeting AgNPs affect the cell cycle and cell division. These experiments demonstrated that nuclear-targeting AgNPs cause DNA double strand breaks and a subsequent increase in the sub G1 (apoptotic) population in our cancer cell model at much lower concentrations than previously reported for nuclear targeting AuNPs. Unlike the M phase accumulation seen in cancer cells treated with AuNPs, an accumulation in the G2 phase of the cell cycle was observed in both cell models when treated with AgNPs. Additionally real-time dark field imaging showed that cancer cells treated with nuclear targeting AgNPs did not undergo cell division and ultimately underwent programmed cell death. A possible explanation of the observed results is discussed in terms of the chemical properties of the nanoparticles. PMID:22010874

  12. Characterization of tumor cells and stem cells by differential nuclear methylation imaging

    NASA Astrophysics Data System (ADS)

    Tajbakhsh, Jian; Wawrowsky, Kolja A.; Gertych, Arkadiusz; Bar-Nur, Ori; Vishnevsky, Eugene; Lindsley, Erik H.; Farkas, Daniel L.

    2008-02-01

    DNA methylation plays a key role in cellular differentiation. Aberrant global methylation patterns are associated with several cancer types, as a result of changes in long-term activation status of up to 50% of genes, including oncogenes and tumor-suppressor genes, which are regulated by methylation and demethylation of promoter region CpG dinucleotides (CpG islands). Furthermore, DNA methylation also occurs in nonisland CpG sites (> 95% of the genome), present once per 80 dinucleotides on average. Nuclear DNA methylation increases during the course of cellular differentiation while cancer cells usually show a net loss in methylation. Given the large dynamic range in DNA methylation load, the methylation pattern of a cell can provide a valuable distinction as to its status during differentiation versus the disease state. By applying immunofluorescence, confocal microscopy and 3D image analysis we assessed the potential of differential nuclear distribution of methylated DNA to be utilized as a biomarker to characterize cells during development and when diseased. There are two major fields that may immediately benefit from this development: (1) the search for factors that contribute to pluripotency and cell fate in human embryonic stem cell expansion and differentiation, and (2) the characterization of tumor cells with regard to their heterogeneity in molecular composition and behavior. We performed topological analysis of the distribution of methylated CpG-sites (MeC) versus heterochromatin. This innovative approach revealed significant differences in colocalization patterns of MeC and heterochromatin-derived signals between undifferentiated and differentiated human embryonic stem cells, as well as untreated AtT20 mouse pituitary tumor cells compared to a subpopulation of these cells treated with 5-azacytidine for 48 hours.

  13. Ryanodine receptors are involved in nuclear calcium oscillation in primary pancreatic {beta}-cells

    SciTech Connect

    Zheng, Ji; Chen, Zheng; Yin, Wenxuan; Miao, Lin; Zhou, Zhansong; Ji, Guangju

    2012-06-29

    Highlights: Black-Right-Pointing-Pointer We found that RyRs are expressed on the nuclear envelope in single primary pancreatic {beta}-cells and isolated nuclei. Black-Right-Pointing-Pointer We showed that the pattern of glucose-induced Ca{sup 2+} oscillation in the nucleus and cytosol was similar. Black-Right-Pointing-Pointer Our results demonstrate that ryanodine-sensitive Ca{sup 2+} stores exist and have function in the pancreatic {beta}-cell nucleus. -- Abstract: Ryanodine receptors (RyRs) are mainly located on the endoplasmic reticulum (ER) and play an important role in regulating glucose-induced cytosolic Ca{sup 2+} oscillation in pancreatic {beta}-cells. However, subcellular locations and functions of RyRs on other cell organelles such as nuclear envelope are not well understood. In order to investigate the role of RyRs in nuclear Ca{sup 2+} oscillation we designed and conducted experiments in intact primary pancreatic {beta}-cells. Immunocytochemistry was used to examine the expression of RYRs on the nuclear envelope. Confocal microscopy was used to evaluate the function of RYRs on the nuclear envelope. We found that RyRs are expressed on the nuclear envelope in single primary pancreatic {beta}-cells and isolated nuclei. Laser scanning confocal microscopy studies indicated that application of glucose to the cells co-incubated with Ca{sup 2+} indicator Fluo-4 AM and cell-permeable nuclear indicator Hoechst 33342 resulted in nuclear Ca{sup 2+} oscillation. The pattern of glucose-induced Ca{sup 2+} oscillation in the nucleus and cytosol was similar. The reduction of Ca{sup 2+} oscillation amplitude by ryanodine was much greater in the nucleus though both the cytosol and the nucleus Ca{sup 2+} amplitude decreased by ryanodine. Our results suggest that functional ryanodine receptors not only exist in endoplasmic reticulum but are also expressed in nuclear envelope of pancreatic {beta}-cells.

  14. Hot microswimmers

    NASA Astrophysics Data System (ADS)

    Kroy, Klaus; Chakraborty, Dipanjan; Cichos, Frank

    2016-11-01

    Hot microswimmers are self-propelled Brownian particles that exploit local heating for their directed self-thermophoretic motion. We provide a pedagogical overview of the key physical mechanisms underlying this promising new technology. It covers the hydrodynamics of swimming, thermophoresis and -osmosis, hot Brownian motion, force-free steering, and dedicated experimental and simulation tools to analyze hot Brownian swimmers.

  15. Hot Flashes

    MedlinePlus

    Diseases and Conditions Hot flashes By Mayo Clinic Staff Hot flashes are sudden feelings of warmth, which are usually most intense over the ... skin may redden, as if you're blushing. Hot flashes can also cause profuse sweating and may ...

  16. Passive permeability and effective pore size of HeLa cell nuclear membranes.

    PubMed

    Samudram, Arunkarthick; Mangalassery, Bijeesh M; Kowshik, Meenal; Patincharath, Nandakumar; Varier, Geetha K

    2016-09-01

    Nuclear pore complexes in the nuclear membrane act as the sole gateway of transport of molecules from the cytoplasm to the nucleus and vice versa. Studies on biomolecular transport through nuclear membranes provide vital data on the nuclear pore complexes. In this work, we use fluorescein isothiocyanate-labeled dextran molecules as a model system and study the passive nuclear import of biomolecules through nuclear pore complexes in digitonin-permeabilized HeLa cells. Experiments are carried out under transient conditions in the time lapse imaging scheme using an in-house constructed confocal laser scanning microscope. Transport rates of dextran molecules having molecular weights of 4-70 kDa corresponding to Stokes radius of 1.4-6 nm are determined. Analyzing the permeability of the nuclear membrane for different sizes the effective pore radius of HeLa cell nuclear membrane is determined to be 5.3 nm, much larger than the value reported earlier using proteins as probe molecules. The range of values reported for the nuclear pore radius suggest that they may not be rigid structures and it is quite probable that the effective pore size of nuclear pore complexes is critically dependent on the probe molecules and on the environmental factors.

  17. Primary biliary cirrhosis and the molecular cell biology of the nuclear envelope.

    PubMed

    Worman, H J

    1994-11-01

    I hope I have demonstrated how basic research on the molecular cell biology of the nuclear envelope has provided information about the autoimmune disease PBC. I have given several examples of how highly specific immunologic reagents, obtained from patients with this disease, have been of value in experiments on the basic cell biology of the nuclear envelope. Continued work should provide further clues on how autoimmunity underlies the pathophysiology of PBC and should also provide additional reagents to study the processes of nuclear protein targeting and cell division.

  18. Generation of embryonic stem cells from mouse adipose-tissue derived cells via somatic cell nuclear transfer

    PubMed Central

    Qin, Yiren; Qin, Jilong; Zhou, Chikai; Li, Jinsong; Gao, Wei-Qiang

    2015-01-01

    Somatic cells can be reprogrammed into embryonic stem cells (ESCs) by nuclear transfer (NT-ESCs), or into induced pluripotent stem cells (iPSCs) by the “Yamanaka method.” However, recent studies have indicated that mouse and human iPSCs are prone to epigenetic and transcriptional aberrations, and that NT-ESCs correspond more closely to ESCs derived from in vitro fertilized embryos than iPSCs. In addition, the procedure of NT-ESCs does not involve gene modification. Demonstration of generation of NT-ESCs using an easily-accessible source of adult cell types would be very important. Adipose tissue is a source of readily accessible donor cells and can be isolated from both males and females at different ages. Here we report that NT-ESCs can be generated from adipose tissue-derived cells (ADCs). At morphological, mRNA and protein levels, these NT-ESCs show classic ESC colonies, exhibit alkaline phosphatase (AP) activity, and display normal diploid karyotypes. Importantly, these cells express pluripotent markers including Oct4, Sox2, Nanog and SSEA-1. Furthermore, they can differentiate in vivo into various types of cells from 3 germinal layers by teratoma formation assays. This study demonstrates for the first time that ESCs can be generated from the adipose tissue by somatic cell nuclear transfer (SCNT) and suggests that ADCs can be a new donor-cell type for potential therapeutic cloning. PMID:25692793

  19. Generation of embryonic stem cells from mouse adipose-tissue derived cells via somatic cell nuclear transfer.

    PubMed

    Qin, Yiren; Qin, Jilong; Zhou, Chikai; Li, Jinsong; Gao, Wei-Qiang

    2015-01-01

    Somatic cells can be reprogrammed into embryonic stem cells (ESCs) by nuclear transfer (NT-ESCs), or into induced pluripotent stem cells (iPSCs) by the "Yamanaka method." However, recent studies have indicated that mouse and human iPSCs are prone to epigenetic and transcriptional aberrations, and that NT-ESCs correspond more closely to ESCs derived from in vitro fertilized embryos than iPSCs. In addition, the procedure of NT-ESCs does not involve gene modification. Demonstration of generation of NT-ESCs using an easily-accessible source of adult cell types would be very important. Adipose tissue is a source of readily accessible donor cells and can be isolated from both males and females at different ages. Here we report that NT-ESCs can be generated from adipose tissue-derived cells (ADCs). At morphological, mRNA and protein levels, these NT-ESCs show classic ESC colonies, exhibit alkaline phosphatase (AP) activity, and display normal diploid karyotypes. Importantly, these cells express pluripotent markers including Oct4, Sox2, Nanog and SSEA-1. Furthermore, they can differentiate in vivo into various types of cells from 3 germinal layers by teratoma formation assays. This study demonstrates for the first time that ESCs can be generated from the adipose tissue by somatic cell nuclear transfer (SCNT) and suggests that ADCs can be a new donor-cell type for potential therapeutic cloning.

  20. Pig transgenesis by piggyBac transposition in combination with somatic cell nuclear transfer.

    PubMed

    Wu, Zhenfang; Xu, Zhiqian; Zou, Xian; Zeng, Fang; Shi, Junsong; Liu, Dewu; Urschitz, Johann; Moisyadi, Stefan; Li, Zicong

    2013-12-01

    The production of animals by somatic cell nuclear transfer (SCNT) is inefficient, with approximately 2% of micromanipulated oocytes going to term and resulting in live births. However, it is the most commonly used method for the generation of cloned transgenic livestock as it facilitates the attainment of transgenic animals once the nuclear donor cells are stably transfected and more importantly as alternatives methods of transgenesis in farm animals have proven even less efficient. Here we describe piggyBac-mediated transposition of a transgene into porcine primary cells and use of these genetically modified cells as nuclear donors for the generation of transgenic pigs by SCNT. Gene transfer by piggyBac transposition serves to provide an alternative approach for the transfection of nuclear donor cells used in SCNT.

  1. Localization of P-glycoprotein at the nuclear envelope of rat brain cells

    SciTech Connect

    Babakhanian, Karlo; Bendayan, Moise; Bendayan, Reina . E-mail: r.bendayan@utoronto.ca

    2007-09-21

    P-Glycoprotein is a plasma membrane drug efflux protein implicated in extrusion of cytotoxic compounds out of a cell. There is now evidence that suggests expression of this transporter at several subcellular sites, including the nucleus, mitochondria, and Golgi apparatus. This study investigated the localization and expression of P-glycoprotein at the nuclear membrane of rat brain microvessel endothelial (RBE4) and microglial (MLS-9) cell lines. Immunocytochemistry at the light and electron microscope levels using P-glycoprotein monoclonals antibodies demonstrated the localization of the protein at the nuclear envelope of RBE4 and MLS-9 cells. Western blot analysis revealed a single band of 170-kDa in purified nuclear membranes prepared from isolated nuclei of RBE4 and MLS-9 cells. These findings indicate that P-glycoprotein is expressed at the nuclear envelope of rat brain cells and suggest a role in multidrug resistance at this subcellular site.

  2. Ceramide-enriched membrane domains in red blood cells and the mechanism of sphingomyelinase-induced hot-cold hemolysis.

    PubMed

    Montes, L-Ruth; López, David J; Sot, Jesús; Bagatolli, Luis A; Stonehouse, Martin J; Vasil, Michael L; Wu, Bill X; Hannun, Yusuf A; Goñi, Félix M; Alonso, Alicia

    2008-10-28

    Hot-cold hemolysis is the phenomenon whereby red blood cells, preincubated at 37 degrees C in the presence of certain agents, undergo rapid hemolysis when transferred to 4 degrees C. The mechanism of this phenomenon is not understood. PlcHR 2, a phospholipase C/sphingomyelinase from Pseudomonas aeruginosa, that is the prototype of a new phosphatase superfamily, induces hot-cold hemolysis. We found that the sphingomyelinase, but not the phospholipase C activity, is essential for hot-cold hemolysis because the phenomenon occurs not only in human erythrocytes that contain both phosphatidylcholine (PC) and sphingomyelin (SM) but also in goat erythrocytes, which lack PC. However, in horse erythrocytes, with a large proportion of PC and almost no SM, hot-cold hemolysis induced by PlcHR 2 is not observed. Fluorescence microscopy observations confirm the formation of ceramide-enriched domains as a result of PlcHR 2 activity. After cooling down to 4 degrees C, the erythrocyte ghost membranes arising from hemolysis contain large, ceramide-rich domains. We suggest that formation of these rigid domains in the originally flexible cell makes it fragile, thus highly susceptible to hemolysis. We also interpret the slow hemolysis observed at 37 degrees C as a phenomenon of gradual release of aqueous contents, induced by the sphingomyelinase activity, as described by Ruiz-Arguello et al. [(1996) J. Biol. Chem. 271, 26616]. These hypotheses are supported by the fact that ceramidase, which is known to facilitate slow hemolysis at 37 degrees C, actually hinders hot-cold hemolysis. Differential scanning calorimetry of erytrocyte membranes treated with PlcHR 2 demonstrates the presence of ceramide-rich domains that are rigid at 4 degrees C but fluid at 37 degrees C. Ceramidase treatment causes the disapperance of the calorimetric signal assigned to ceramide-rich domains. Finally, in liposomes composed of SM, PC, and cholesterol, which exhibit slow release of aqueous contents at 37

  3. Role of Actin Filaments in Correlating Nuclear Shape and Cell Spreading

    PubMed Central

    Vishavkarma, Renu; Raghavan, Swetavalli; Kuyyamudi, Chandrashekar; Majumder, Abhijit; Dhawan, Jyotsna; Pullarkat, Pramod A.

    2014-01-01

    It is well known that substrate properties like stiffness and adhesivity influence stem cell morphology and differentiation. Recent experiments show that cell morphology influences nuclear geometry and hence gene expression profile. The mechanism by which surface properties regulate cell and nuclear properties is only beginning to be understood. Direct transmission of forces as well as chemical signalling are involved in this process. Here, we investigate the formal aspect by studying the correlation between cell spreading and nuclear deformation using Mesenchymal stem cells under a wide variety of conditions. It is observed that a robust quantitative relation holds between the cell and nuclear projected areas, irrespective of how the cell area is modified or when various cytoskeletal or nuclear components are perturbed. By studying the role of actin stress fibers in compressing the nucleus we propose that nuclear compression by stress fibers can lead to enhanced cell spreading due to an interplay between elastic and adhesion factors. The significance of myosin-II in regulating this process is also explored. We demonstrate this effect using a simple technique to apply external compressive loads on the nucleus. PMID:25251154

  4. Nuclear but Not Cytosolic Phosphoinositide 3-Kinase Beta Has an Essential Function in Cell Survival ▿

    PubMed Central

    Kumar, Amit; Redondo-Muñoz, Javier; Perez-García, Vicente; Cortes, Isabel; Chagoyen, Monica; Carrera, Ana C.

    2011-01-01

    Class IA phosphoinositide 3-kinases (PI3Ks) are heterodimeric enzymes composed of a p85 regulatory and a p110 catalytic subunit that induce the formation of 3-polyphosphoinositides, which mediate cell survival, division, and migration. There are two ubiquitous PI3K isoforms p110α and p110β that have nonredundant functions in embryonic development and cell division. However, whereas p110α concentrates in the cytoplasm, p110β localizes to the nucleus and modulates nuclear processes such as DNA replication and repair. At present, the structural features that determine p110β nuclear localization remain unknown. We describe here that association with the p85β regulatory subunit controls p110β nuclear localization. We identified a nuclear localization signal (NLS) in p110β C2 domain that mediates its nuclear entry, as well as a nuclear export sequence (NES) in p85β. Deletion of p110β induced apoptosis, and complementation with the cytoplasmic C2-NLS p110β mutant was unable to restore cell survival. These studies show that p110β NLS and p85β NES regulate p85β/p110β nuclear localization, supporting the idea that nuclear, but not cytoplasmic, p110β controls cell survival. PMID:21383062

  5. Photoproduct frequency is not the major determinant of UV base substitution hot spots or cold spots in human cells

    SciTech Connect

    Brash, D.E.; Seetharam, S.; Kraemer, K.H.; Seidman, M.M.; Bredberg, A.

    1987-06-01

    The role of UV radiation-induced photoproducts in initiating base substitution mutations in human cells was examined by measuring photoproduct frequency distributions and mutations in a supF tRNA gene on a shuttle vector plasmid transfected into DNA repair-deficient cells (xeroderma pigmentosum, complementation group A) and into normal cells. Frequencies of cyclobutane dimers and pyrimidine-pyrimidone (6-4) photoproducts varied by as much as 80-fold at different dipyrimidine sites within the gene. All transition mutations occurred at dipyrimidine sites, predominantly at cytosine, with a 17-fold variation in mutation frequency between different sites. Removal of greater than 99% of the cyclobutane dimers by in vitro photoreactivation before transfection reduced the mutation frequency while preserving the mutation distribution, indicating that (i) cytosine-containing cyclobutane dimers were the major mutagenic lesions at these sites and (ii) cytosine-containing non-cyclobutane dimer photoproducts were also mutagenic lesions. However, at individual dipyrimidine sites neither the frequency of cyclobutane dimers nor the frequency of pyrimidine-pyrimidone (6-4) photoproducts correlated with the mutation frequency, even in the absence of excision repair. Mutation hot spots occurred at sites with low or high frequency of photoproduct formation and mutation cold spots occurred at sites with many photoproducts. These results suggest that although photoproducts are required for UV mutagenesis, the prominence of most mutation hot spots and cold spots is primarily determined by DNA structural features rather than by the frequency of DNA photoproducts.

  6. Evaluation of porcine stem cell competence for somatic cell nuclear transfer and production of cloned animals.

    PubMed

    Secher, Jan O; Liu, Ying; Petkov, Stoyan; Luo, Yonglun; Li, Dong; Hall, Vanessa J; Schmidt, Mette; Callesen, Henrik; Bentzon, Jacob F; Sørensen, Charlotte B; Freude, Kristine K; Hyttel, Poul

    2017-03-01

    Porcine somatic cell nuclear transfer (SCNT) has been used extensively to create genetically modified pigs, but the efficiency of the methodology is still low. It has been hypothesized that pluripotent or multipotent stem cells might result in increased SCNT efficacy as these cells are closer than somatic cells to the epigenetic state found in the blastomeres and therefore need less reprogramming. Our group has worked with porcine SCNT during the last 20 years and here we describe our experience with SCNT of 3 different stem cell lines. The porcine stem cells used were: Induced pluripotent stem cells (iPSCs) created by lentiviral doxycycline-dependent reprogramming and cultered with a GSK3β- and MEK-inhibitor (2i) and leukemia inhibitor factor (LIF) (2i LIF DOX-iPSCs), iPSCs created by a plasmid-based reprogramming and cultured with 2i and fibroblast growth factor (FGF) (2i FGF Pl-iPSCs) and embryonic germ cells (EGCs), which have earlier been characterized as being multipotent. The SCNT efficiencies of these stem cell lines were compared with that of the two fibroblast cell lines from which the iPSC lines were derived. The blastocyst rates for the 2i LIF DOX-iPSCs were 14.7%, for the 2i FGF Pl-iPSC 10.1%, and for the EGCs 34.5% compared with the fibroblast lines yielding 36.7% and 25.2%. The fibroblast- and EGC-derived embryos were used for embryo transfer and produced live offspring at similar low rates of efficiency (3.2 and 4.0%, respectively) and with several instances of malformations. In conclusion, potentially pluripotent porcine stem cells resulted in lower rates of embryonic development upon SCNT than multipotent stem cells and differentiated somatic cells.

  7. Limited expression of nuclear pore membrane glycoprotein 210 in cell lines and tissues suggests cell-type specific nuclear pores in metazoans.

    PubMed

    Olsson, Magnus; Schéele, Susanne; Ekblom, Peter

    2004-01-15

    The nuclear pore complex (NPC) is the only known gateway for nucleocytoplasmic traffic. The nuclear pore membrane glycoprotein 210 (POM210/gp210) is considered to be important for the assembly and structure of pore complexes in metazoan cells. However, here we demonstrate cell-type specific expression of the gp210 protein during mouse organogenesis. As shown previously for its mRNA, distinct expression of the gp210 was seen in developing epithelia and some other cell types, whereas it was undetectable in nuclei of several other embryonic tissue compartments. In sharp contrast, monoclonal antibody 414 recognizing four non-membrane nucleoporins, stained the nuclear envelope of all cell types. In four cultured mouse cell lines, gp210 mRNA and protein were below detection levels, in contrast to some other nucleoporins tested. Distinct expression of gp210 mRNA and protein was seen in cultured mouse embryonic stem (ES) cells. These findings support the view of cell-type specific NPCs in metazoans and that the gp210 gene is regulated by cell-type specific control elements not shared by other nucleoporins. Although it cannot be excluded that very low expression levels of gp210 are sufficient to allow attachment of NPCs, a more likely alternative is that it has cell-type specific functions.

  8. Changes in plant cell-wall structure of corn stover due to hot compressed water pretreatment and enhanced enzymatic hydrolysis.

    PubMed

    Zhou, Wei; Yang, Maohua; Wang, Caixia; Liu, Jianfei; Xing, Jianmin

    2014-08-01

    Corn stover is a potential feedstock for biofuel production. This work investigated physical and chemical changes in plant cell-wall structure of corn stover due to hot compressed water (HCW) pretreatment at 170-190 °C in a tube reactor. Chemical composition analysis showed the soluble hemicellulose content increased with pretreatment temperature, whereas the hemicellulose content decreased from 29 to 7 % in pretreated solids. Scanning electron microscopy revealed the parenchyma-type second cell-wall structure of the plant was almost completely removed at 185 °C, and the sclerenchyma-type second cell wall was greatly damaged upon addition of 5 mmol/L ammonium sulfate during HCW pretreatment. These changes favored accessibility for enzymatic action. Enzyme saccharification of solids by optimized pretreatment with HCW at 185 °C resulted in an enzymatic hydrolysis yield of 87 %, an enhancement of 77 % compared to the yield from untreated corn stover.

  9. The ability of mouse nuclear transfer embryonic stem cells to differentiate into primordial germ cells.

    PubMed

    Mansouri, Vahid; Salehi, Mohammad; Nourozian, Mohsen; Fadaei, Fatemeh; Farahani, Reza Mastery; Piryaei, Abbas; Delbari, Ali

    2015-05-01

    Nuclear transfer embryonic stem cells (ntESCs) show stem cell characteristics such as pluripotency but cause no immunological disorders. Although ntESCs are able to differentiate into somatic cells, the ability of ntESCs to differentiate into primordial germ cells (PGCs) has not been examined. In this work, we examined the capacity of mouse ntESCs to differentiate into PGCs in vitro. ntESCs aggregated to form embryoid bodies (EB) in EB culture medium supplemented with bone morphogenetic protein 4(BMP4) as the differentiation factor. The expression level of specific PGC genes was compared at days 4 and 8 using real time PCR. Flow cytometry and immunocytochemical staining were used to detect Mvh as a specific PGC marker. ntESCs expressed particular genes related to different stages of PGC development. Flow cytometry and immunocytochemical staining confirmed the presence of Mvh protein in a small number of cells. There were significant differences between cells that differentiated into PGCs in the group treated with Bmp4 compared to non-treated cells. These findings indicate that ntESCs can differentiate into putative PGCs. Improvement of ntESC differentiation into PGCs may be a reliable means of producing mature germ cells.

  10. Potential of primary kidney cells for somatic cell nuclear transfer mediated transgenesis in pig

    PubMed Central

    2012-01-01

    Background Somatic cell nuclear transfer (SCNT) is currently the most efficient and precise method to generate genetically tailored pig models for biomedical research. However, the efficiency of this approach is crucially dependent on the source of nuclear donor cells. In this study, we evaluate the potential of primary porcine kidney cells (PKCs) as cell source for SCNT, including their proliferation capacity, transfection efficiency, and capacity to support full term development of SCNT embryos after additive gene transfer or homologous recombination. Results PKCs could be maintained in culture with stable karyotype for up to 71 passages, whereas porcine fetal fibroblasts (PFFs) and porcine ear fibroblasts (PEFs) could be hardly passaged more than 20 times. Compared with PFFs and PEFs, PKCs exhibited a higher proliferation rate and resulted in a 2-fold higher blastocyst rate after SCNT and in vitro cultivation. Among the four transfection methods tested with a GFP expression plasmid, best results were obtained with the NucleofectorTM technology, resulting in transfection efficiencies of 70% to 89% with high fluorescence intensity, low cytotoxicity, good cell proliferation, and almost no morphological signs of cell stress. Usage of genetically modified PKCs in SCNT resulted in approximately 150 piglets carrying at least one of 18 different transgenes. Several of those pigs originated from PKCs that underwent homologous recombination and antibiotic selection before SCNT. Conclusion The high proliferation capacity of PKCs facilitates the introduction of precise and complex genetic modifications in vitro. PKCs are thus a valuable cell source for the generation of porcine biomedical models by SCNT. PMID:23140586

  11. AFC-1 Transmutation Fuels Post-Irradiation Hot Cell Examination 4-8 at.% - Final Report (Irradiation Experiments AFC-1B, -1F and -1Æ)

    SciTech Connect

    Bruce Hilton; Douglas Porter; Steven Hayes

    2006-09-01

    The AFC-1B, AFC-1F and AFC-1Æ irradiation tests are part of a series of test irradiations designed to evaluate the feasibility of the use of actinide bearing fuel forms in advanced fuel cycles for the transmutation of transuranic elements from nuclear waste. The tests were irradiated in the Idaho National Laboratory’s (INL) Advanced Test Reactor (ATR) to an intermediate burnup of 4 to 8 at% (2.7 - 6.8 x 1020 fiss/cm3). The tests contain metallic and nitride fuel forms with non-fertile (i.e., no uranium) and low-fertile (i.e., uranium bearing) compositions. Results of postirradiation hot cell examinations of AFC-1 irradiation tests are reported for eleven metallic alloy transmutation fuel rodlets and five nitride transmutation fuel rodlets. Non-destructive examinations included visual examination, dimensional inspection, gamma scan analysis, and neutron radiography. Detailed examinations, including fission gas puncture and analysis, metallography / ceramography and isotopics and burnup analyses, were performed on five metallic alloy and three nitride transmutation fuels. Fuel performance of both metallic alloy and nitride fuel forms was best correlated with fission density as a burnup metric rather than at.% depletion. The actinide bearing transmutation metallic alloy compositions exhibit irradiation performance very similar to U-xPu-10Zr fuel at equivalent fission densities. The irradiation performance of nitride transmutation fuels was comparable to limited data published on mixed nitride systems.

  12. Nuclear physics: quantitative single-cell approaches to nuclear organization and gene expression.

    PubMed

    Lionnet, T; Wu, B; Grünwald, D; Singer, R H; Larson, D R

    2010-01-01

    The internal workings of the nucleus remain a mystery. A list of component parts exists, and in many cases their functional roles are known for events such as transcription, RNA processing, or nuclear export. Some of these components exhibit structural features in the nucleus, regions of concentration or bodies that have given rise to the concept of functional compartmentalization--that there are underlying organizational principles to be described. In contrast, a picture is emerging in which transcription appears to drive the assembly of the functional components required for gene expression, drawing from pools of excess factors. Unifying this seemingly dual nature requires a more rigorous approach, one in which components are tracked in time and space and correlated with onset of specific nuclear functions. In this chapter, we anticipate tools that will address these questions and provide the missing kinetics of nuclear function. These tools are based on analyzing the fluctuations inherent in the weak signals of endogenous nuclear processes and determining values for them. In this way, it will be possible eventually to provide a computational model describing the functional relationships of essential components.

  13. Expression of nuclear membrane proteins in normal, hyperplastic, and neoplastic thyroid epithelial cells.

    PubMed

    Wang, Jieying; Kondo, Tetsuo; Yamane, Tetsu; Nakazawa, Tadao; Oish, Naoki; Mochizuki, Kunio; Katoh, Ryohei

    2015-10-01

    Emerin, lamin A/C, lamin B, and lamin-associated polypeptide 2 (LAP2) are nuclear membrane proteins that play an important role in maintaining nuclear structure and coordinating cell activity. We studied the expression and significance of nuclear membrane proteins in neoplastic thyroid cells by immunohistochemistry, RT-PCR, and real-time PCR. In papillary carcinomas (PCs), the nuclear proteins most frequently expressed at high levels were emerin (82 % positive), lamin A/C (64 %), and LAP2 (82 %). Follicular carcinomas (FCs) most frequently expressed lamin B, while none of the undifferentiated carcinomas (UCs) showed strong expression of emerin or lamin A/C. In all medullary carcinomas (MCs), intermediate to high levels of expression of lamin A/C and LAP2 were found. By RT-PCR analysis, messenger RNA (mRNA) expression of all nuclear membrane proteins except emerin was higher in PC than in normal tissue. Real-time PCR analysis showed that mRNA expression of nuclear membrane protein varied between cell lines. Our findings suggest that expression of nuclear membrane proteins may be related to follicular function in normal and hyperplastic follicles, and we hypothesize that they are also involved in the proliferation and differentiation of neoplastic thyroid cells. We suggest that they reflect the biological nature and/or function of normal, hyperplastic, and neoplastic thyroid cells and may have some value in diagnosing thyroid tumors.

  14. Efficient selection of Gal-knockout pig cells for somatic cell nuclear transfer.

    PubMed

    Reyes, Luz M; Estrada, Jose L; Ivary, Bess; Sidner, Richard A; Paris, Leela L; Tector, A Joseph

    2013-10-01

    The process of selecting transgenic cells has been one of the bottlenecks in the generation of transgenic animals by somatic cell nuclear transfer (SCNT). In particular, selection for the Gal double-knockout (Gal-DKO) genotype has been time consuming and inefficient. The objective of this work was to generate a highly efficient system to select Gal-DKO cells to be used in SCNT without affecting the efficiency in production of Gal-null pigs. Fetal liver-derived cells deficient in Gal-expression were initially selected by fluorescence-activated cell sorting (FACS) using IB4 conjugated to a fluorescent dye. Cells recovered by FACS were cultured and expanded, followed by a second round of selection using streptavidin magnetic beads and IB4 lectin biotin. Recovery efficiency of target cells was 0.04% for the first selection using FACS and 0.3% for the second round by magnetic beads. Full reprogramming was obtained on selected Gal-DKO cells after FACS and magnetic beads selection, when used for SCNT to produce the Gal-null piglets. Cells obtained from magnetic beads developed 48 colonies; the Gal-null genotype was found in 44 of them (91.7%). Three of these colonies were used to generate piglets by SCNT. From three recipients receiving embryos, two became pregnant and produced 17 piglets, all of them DKO. Sequential selection of Gal-DKO cells by FACS/magnetic beads is a highly efficient system to generate null cells. Selected cells were successfully used to generate healthy double-knockout piglets by SCNT. Copyright © 2013 Elsevier Inc. All rights reserved.

  15. Intracellular lysyl oxidase: Effect of a specific inhibitor on nuclear mass in proliferating cells

    SciTech Connect

    Saad, Fawzy A.; Torres, Marie; Wang, Hao; Graham, Lila

    2010-06-11

    LOX, the principal enzyme involved in crosslinking of collagen, was the first of several lysyl oxidase isotypes to be characterized. Its active form was believed to be exclusively extracellular. Active LOX was later reported to be present in cell nuclei; its function there is unknown. LOX expression opposes the effect of mutationally activated Ras, which is present in about 30% of human cancers. The mechanism of LOX in countering the action of Ras is also unknown. In the present work, assessment of nuclear protein for possible effects of lysyl oxidase activity led to the discovery that proliferating cells dramatically increase their nuclear protein content when exposed to BAPN ({beta}-aminopropionitrile), a highly specific lysyl oxidase inhibitor that reportedly blocks LOX inhibition of Ras-induced oocyte maturation. In three cell types (PC12 cells, A7r5 smooth muscle cells, and NIH 3T3 fibroblasts), BAPN caused a 1.8-, 1.7-, and 2.1-fold increase in total nuclear protein per cell, respectively, affecting all major components in both nuclear matrix and chromatin fractions. Since nuclear size is correlated with proliferative status, enzyme activity restricting nuclear growth may be involved in the lysyl oxidase tumor suppressive effect. Evidence is also presented for the presence of apparent lysyl oxidase isotype(s) containing a highly conserved LOX active site sequence in the nuclei of PC12 cells, which do not manufacture extracellular lysyl oxidase substrates. Results reported here support the hypothesis that nuclear lysyl oxidase regulates nuclear growth, and thereby modulates cell proliferation.

  16. cdc25 is a nuclear protein expressed constitutively throughout the cell cycle in nontransformed mammalian cells

    PubMed Central

    1992-01-01

    A family of proteins homologous to the cdc25 gene product of the fission yeast bear specific protein tyrosine phosphatase activity involved in the activation of the p34cdc2-cyclin B kinase. Using affinity-purified antibodies raised against a synthetic peptide corresponding to the catalytic site of the cdc25 phosphatase, we show that cdc25 protein is constitutively expressed throughout the cell cycle of nontransformed mammalian fibroblasts and does not undergo major changes in protein level. By indirect immunofluorescence, cdc25 protein is found essentially localized in the nucleus throughout interphase and during early prophase. Just before the complete nuclear envelope breakdown at the prophase-prometaphase boundary, cdc25 proteins are redistributed throughout the cytoplasm. During metaphase and anaphase, cdc25 staining remains distributed throughout the cell and excludes the condensed chromosomes. The nuclear locale reappears during telophase. In light of the recent data describing the cytoplasmic localization of cyclin B protein (Pines, J., and T. Hunter. 1991. J. Cell Biol. 115:1-17), the data presented here suggest that separation in two distinct cellular compartments of the cdc25 phosphatase and its substrate p34cdc2-cyclin B may be of importance in the regulation of the cdc2 kinase activity. PMID:1500423

  17. A hot-electron thermophotonic solar cell demonstrated by thermal up-conversion of sub-bandgap photons

    PubMed Central

    Farrell, Daniel J.; Sodabanlu, Hassanet; Wang, Yunpeng; Sugiyama, Masakazu; Okada, Yoshitaka

    2015-01-01

    The direct conversion of solar energy to electricity can be broadly separated into two main categories: photovoltaics and thermal photovoltaics, where the former utilizes gradients in electrical potential and the latter thermal gradients. Conventional thermal photovoltaics has a high theoretical efficiency limit (84%) but in practice cannot be easily miniaturized and is limited by the engineering challenges of sustaining large (>1,000 K) temperature gradients. Here we show a hot-carrier-based thermophotonic solar cell, which combines the compact nature of photovoltaic devices with the potential to reach the high-efficiency regime of thermal photovoltaics. In the device, a thermal gradient of 500 K is established by hot electrons, under Stokes illumination, rather than by raising the temperature of the material itself. Under anti-Stokes (sub-bandgap) illumination we observe a thermal gradient of ∼20 K, which is maintained by steady-state Auger heating of carriers and corresponds to a internal thermal up-conversion efficiency of 30% between the collector and solar cell. PMID:26541415

  18. A hot-electron thermophotonic solar cell demonstrated by thermal up-conversion of sub-bandgap photons.

    PubMed

    Farrell, Daniel J; Sodabanlu, Hassanet; Wang, Yunpeng; Sugiyama, Masakazu; Okada, Yoshitaka

    2015-11-06

    The direct conversion of solar energy to electricity can be broadly separated into two main categories: photovoltaics and thermal photovoltaics, where the former utilizes gradients in electrical potential and the latter thermal gradients. Conventional thermal photovoltaics has a high theoretical efficiency limit (84%) but in practice cannot be easily miniaturized and is limited by the engineering challenges of sustaining large (>1,000 K) temperature gradients. Here we show a hot-carrier-based thermophotonic solar cell, which combines the compact nature of photovoltaic devices with the potential to reach the high-efficiency regime of thermal photovoltaics. In the device, a thermal gradient of 500 K is established by hot electrons, under Stokes illumination, rather than by raising the temperature of the material itself. Under anti-Stokes (sub-bandgap) illumination we observe a thermal gradient of ∼20 K, which is maintained by steady-state Auger heating of carriers and corresponds to a internal thermal up-conversion efficiency of 30% between the collector and solar cell.

  19. A hot-electron thermophotonic solar cell demonstrated by thermal up-conversion of sub-bandgap photons

    NASA Astrophysics Data System (ADS)

    Farrell, Daniel J.; Sodabanlu, Hassanet; Wang, Yunpeng; Sugiyama, Masakazu; Okada, Yoshitaka

    2015-11-01

    The direct conversion of solar energy to electricity can be broadly separated into two main categories: photovoltaics and thermal photovoltaics, where the former utilizes gradients in electrical potential and the latter thermal gradients. Conventional thermal photovoltaics has a high theoretical efficiency limit (84%) but in practice cannot be easily miniaturized and is limited by the engineering challenges of sustaining large (>1,000 K) temperature gradients. Here we show a hot-carrier-based thermophotonic solar cell, which combines the compact nature of photovoltaic devices with the potential to reach the high-efficiency regime of thermal photovoltaics. In the device, a thermal gradient of 500 K is established by hot electrons, under Stokes illumination, rather than by raising the temperature of the material itself. Under anti-Stokes (sub-bandgap) illumination we observe a thermal gradient of ~20 K, which is maintained by steady-state Auger heating of carriers and corresponds to a internal thermal up-conversion efficiency of 30% between the collector and solar cell.

  20. A comparison between nuclear dismantling during plant and animal programmed cell death.

    PubMed

    Domínguez, Fernando; Cejudo, Francisco Javier

    2012-12-01

    Programmed cell death (PCD) is a process of organized destruction of cells, essential for the development and maintenance of cellular homeostasis of multicellular organisms. Cells undergoing PCD begin a degenerative process in response to internal or external signals, whereby the nucleus becomes one of the targets. The process of nuclear dismantling includes events affecting the nuclear envelope, such as formation of lobes at the nuclear surface, selective proteolysis of nucleoporins and nuclear pore complex clustering. In addition, chromatin condensation increases in coordination with DNA fragmentation. These processes have been largely studied in animals, but remain poorly understood in plants. The overall process of cell death has different morphological and biochemical features in plants and animals. However, recent advances suggest that nuclear dismantling in plant cells progresses with morphological and biochemical characteristics similar to those in apoptotic animal cells. In this review, we summarize nuclear dismantling in plant PCD, focusing on the similarities and differences with their animal counterparts. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  1. The effect of nanoparticle size and NLS density on nuclear targeting in cancer and normal cells; impaired nuclear import and aberrant nanoparticle intracellular trafficking in glioma.

    PubMed

    Tammam, Salma N; Azzazy, Hassan M E; Lamprecht, Alf

    2017-02-27

    The cell nucleus is an interesting target in many diseases with particular interest in cancer. Previously, nuclear targeted small and large chitosan nanoparticles (S-NPs≈25nm, and L-NPs≈150nm respectively), modified with low, intermediate and high densities of NLS (L-NLS, I-NLS and H-NLS) were developed and assessed in L929 fibroblasts. However, to evade apoptosis and stimulate tumor growth cancer cells are capable of manipulating the nuclear-cytoplasmic transport on many levels, making NPs that are capable of nuclear targeting in normal cells incapable of doing so in cancer. For such reason, here, the nuclear delivery efficiency of S-NPs and L-NPs was assessed as a function of their NLS density in cancer and non-cancer cells. For S-NPs, in all cells tested, NLS was unnecessary for nuclear delivery; unmodified S-NPs showed higher nuclear delivery than NLS-S-NPs due to their ability to gain nuclear entry in a passive manner. For L-NPs, L-NLS-L-NPs showed ≈ 8.5, 33, 1.8 and 7.2 fold higher nuclear deliveries than H-NLS-L-NPs in L929 fibroblasts, primary human fibroblasts, HEK 293 and lung cancer cells, respectively. In glioma however, unmodified L-NPs showed highest nuclear delivery, whereas NLS-L-NPs were retained in the cytoplasm. Experiments conducted in the presence of inhibitors of the classical nuclear import pathway indicated that due to overexpression of importin α, classical nuclear import in glioma is impaired leading to aberrant NP intracellular trafficking and nuclear import.

  2. Assessing mRNA nuclear export in mammalian cells by microinjection.

    PubMed

    Lee, Eliza S; Palazzo, Alexander F

    2017-08-15

    The nuclear export of mRNAs is an important yet little understood part of eukaryotic gene expression. One of the easiest methods for monitoring mRNA export in mammalian tissue culture cells is through the microinjection of DNA plasmids into the nucleus and monitoring the distribution of the transcribed product over time. Here we describe how to setup a microscope equipped with a micromanipulator used in cell microinjections, and we explain how to perform a nuclear mRNA export assay and obtain the nuclear export rate for any given mRNA. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Nuclear export of the glucocorticoid receptor is accelerated by cell fusion-dependent release of calreticulin.

    PubMed

    Walther, Rhian F; Lamprecht, Claudia; Ridsdale, Andrew; Groulx, Isabelle; Lee, Stephen; Lefebvre, Yvonne A; Haché, Robert J G

    2003-09-26

    Nucleocytoplasmic exchange of nuclear hormone receptors is hypothesized to allow for rapid and direct interactions with cytoplasmic signaling factors. In addition to recycling between a naïve, chaperone-associated cytoplasmic complex and a liganded chaperone-free nuclear form, the glucocorticoid receptor (GR) has been observed to shuttle between nucleus and cytoplasm. Nuclear export of GR and other nuclear receptors has been proposed to depend on direct interactions with calreticulin, which is predominantly localized to the lumen of the endoplasmic reticulum. We show that rapid calreticulin-mediated nuclear export of GR is a specific response to transient disruption of the endoplasmic reticulum that occurs during polyethylene glycol-mediated cell fusion. Using live and digitonin-permeabilized cells we demonstrate that, in the absence of cell fusion, GR nuclear export occurs slowly over a period of many hours independent of direct interaction with calreticulin. Our findings temper expectations that nuclear receptors respond rapidly and directly to cytoplasmic signals in the absence of additional regulatory control. These results highlight the importance of verifying findings of nucleocytoplasmic trafficking using techniques in addition to heterokaryon cell fusion.

  4. Decontamination of hot cells K-1, K-3, M-1, M-3, and A-1, M-Wing, Building 200: Project final report Argonne National Laboratory-East

    SciTech Connect

    Cheever, C.L.; Rose, R.W.

    1996-09-01

    The purpose of this project was to remove radioactively contaminated materials and equipment from the hot cells, to decontaminate the hot cells, and to dispose of the radioactive waste. The goal was to reduce stack releases of Rn-220 and to place the hot cells in an emptied, decontaminated condition with less than 10 {micro}Sv/h (1 mrem/h) general radiation background. The following actions were needed: organize and mobilize a decontamination team; prepare decontamination plans and procedures; perform safety analyses to ensure protection of the workers, public, and environment; remotely size-reduce, package, and remove radioactive materials and equipment for waste disposal; remotely decontaminate surfaces to reduce hot cell radiation background levels to allow personnel entries using supplied air and full protective suits; disassemble and package the remaining radioactive materials and equipment using hands-on techniques; decontaminate hot cell surfaces to remove loose radioactive contaminants and to attain a less than 10 {micro}Sv/h (1 mrem/h) general background level; document and dispose of the radioactive and mixed waste; and conduct a final radiological survey.

  5. Nuclear envelope defects cause stem cell dysfunction in premature-aging mice

    PubMed Central

    Espada, Jesús; Varela, Ignacio; Flores, Ignacio; Ugalde, Alejandro P.; Cadiñanos, Juan; Pendás, Alberto M.; Stewart, Colin L.; Tryggvason, Karl; Blasco, María A.; Freije, José M.P.; López-Otín, Carlos

    2008-01-01

    Nuclear lamina alterations occur in physiological aging and in premature aging syndromes. Because aging is also associated with abnormal stem cell homeostasis, we hypothesize that nuclear envelope alterations could have an important impact on stem cell compartments. To evaluate this hypothesis, we examined the number and functional competence of stem cells in Zmpste24-null progeroid mice, which exhibit nuclear lamina defects. We show that Zmpste24 deficiency causes an alteration in the number and proliferative capacity of epidermal stem cells. These changes are associated with an aberrant nuclear architecture of bulge cells and an increase in apoptosis of their supporting cells in the hair bulb region. These alterations are rescued in Zmpste24−/−Lmna+/− mutant mice, which do not manifest progeroid symptoms. We also report that molecular signaling pathways implicated in the regulation of stem cell behavior, such as Wnt and microphthalmia transcription factor, are altered in Zmpste24−/− mice. These findings establish a link between age-related nuclear envelope defects and stem cell dysfunction. PMID:18378773

  6. Increased mechanosensitivity and nuclear stiffness in Hutchinson-Gilford progeria cells: effects of farnesyltransferase inhibitors.

    PubMed

    Verstraeten, Valerie L R M; Ji, Julie Y; Cummings, Kiersten S; Lee, Richard T; Lammerding, Jan

    2008-06-01

    Hutchinson-Gilford progeria syndrome (HGPS), reportedly a model for normal aging, is a genetic disorder in children marked by dramatic signs suggestive for premature aging. It is usually caused by de novo mutations in the nuclear envelope protein lamin A. Lamins are essential to maintaining nuclear integrity, and loss of lamin A/C results in increased cellular sensitivity to mechanical strain and defective mechanotransduction signaling. Since increased mechanical sensitivity in vascular cells could contribute to loss of smooth muscle cells and the development of arteriosclerosis--the leading cause of death in HGPS patients--we investigated the effect of mechanical stress on cells from HGPS patients. We found that skin fibroblasts from HGPS patients developed progressively stiffer nuclei with increasing passage number. Importantly, fibroblasts from HGPS patients had decreased viability and increased apoptosis under repetitive mechanical strain, as well as attenuated wound healing, and these defects preceded changes in nuclear stiffness. Treating fibroblasts with farnesyltransferase inhibitors restored nuclear stiffness in HGPS cells and accelerated the wound healing response in HGPS and healthy control cells by increasing the directional persistence of migrating cells. However, farnesyltransferase inhibitors did not improve cellular sensitivity to mechanical strain. These data suggest that increased mechanical sensitivity in HGPS cells is unrelated to changes in nuclear stiffness and that increased biomechanical sensitivity could provide a potential mechanism for the progressive loss of vascular smooth muscle cells under physiological strain in HGPS patients.

  7. A Chemomechanical Model for Nuclear Morphology and Stresses during Cell Transendothelial Migration.

    PubMed

    Cao, Xuan; Moeendarbary, Emad; Isermann, Philipp; Davidson, Patricia M; Wang, Xiao; Chen, Michelle B; Burkart, Anya K; Lammerding, Jan; Kamm, Roger D; Shenoy, Vivek B

    2016-10-04

    It is now evident that the cell nucleus undergoes dramatic shape changes during important cellular processes such as cell transmigration through extracellular matrix and endothelium. Recent experimental data suggest that during cell transmigration the deformability of the nucleus could be a limiting factor, and the morphological and structural alterations that the nucleus encounters can perturb genomic organization that in turn influences cellular behavior. Despite its importance, a biophysical model that connects the experimentally observed nuclear morphological changes to the underlying biophysical factors during transmigration through small constrictions is still lacking. Here, we developed a universal chemomechanical model that describes nuclear strains and shapes and predicts thresholds for the rupture of the nuclear envelope and for nuclear plastic deformation during transmigration through small constrictions. The model includes actin contraction and cytosolic back pressure that squeeze the nucleus through constrictions and overcome the mechanical resistance from deformation of the nucleus and the constrictions. The nucleus is treated as an elastic shell encompassing a poroelastic material representing the nuclear envelope and inner nucleoplasm, respectively. Tuning the chemomechanical parameters of different components such as cell contractility and nuclear and matrix stiffnesses, our model predicts the lower bounds of constriction size for successful transmigration. Furthermore, treating the chromatin as a plastic material, our model faithfully reproduced the experimentally observed irreversible nuclear deformations after transmigration in lamin-A/C-deficient cells, whereas the wild-type cells show much less plastic deformation. Along with making testable predictions, which are in accord with our experiments and existing literature, our work provides a realistic framework to assess the biophysical modulators of nuclear deformation during cell transmigration.

  8. Control of hot-carrier relaxation for realizing ideal quantum-dot intermediate-band solar cells.

    PubMed

    Tex, David M; Kamiya, Itaru; Kanemitsu, Yoshihiko

    2014-02-18

    For intermediate-band solar cells, the broad absorption spectrum of quantum dots (QDs) offers a favorable conversion efficiency, and photocurrent generation via efficient two-step two-photon-absorption (TS-TPA) in QDs is essential for realizing high-performance solar cells. In the last decade, many works were dedicated to improve the TS-TPA efficiency by modifying the QD itself, however, the obtained results are far from the requirements for practical applications. To reveal the mechanisms behind the low TS-TPA efficiency in QDs, we report here on two- and three-beam photocurrent measurements of InAs quantum structures embedded in AlGaAs. Comparison of two- and three-beam photocurrent spectra obtained by subbandgap excitation reveals that the QD TS-TPA efficiency is improved significantly by suppressing the relaxation of hot TS-TPA carriers to unoccupied shallow InAs quantum structure states.

  9. Control of hot-carrier relaxation for realizing ideal quantum-dot intermediate-band solar cells

    NASA Astrophysics Data System (ADS)

    Tex, David M.; Kamiya, Itaru; Kanemitsu, Yoshihiko

    2014-02-01

    For intermediate-band solar cells, the broad absorption spectrum of quantum dots (QDs) offers a favorable conversion efficiency, and photocurrent generation via efficient two-step two-photon-absorption (TS-TPA) in QDs is essential for realizing high-performance solar cells. In the last decade, many works were dedicated to improve the TS-TPA efficiency by modifying the QD itself, however, the obtained results are far from the requirements for practical applications. To reveal the mechanisms behind the low TS-TPA efficiency in QDs, we report here on two- and three-beam photocurrent measurements of InAs quantum structures embedded in AlGaAs. Comparison of two- and three-beam photocurrent spectra obtained by subbandgap excitation reveals that the QD TS-TPA efficiency is improved significantly by suppressing the relaxation of hot TS-TPA carriers to unoccupied shallow InAs quantum structure states.

  10. Histone modification of embryonic stem cells produced by somatic cell nuclear transfer and fertilized blastocysts.

    PubMed

    Farifteh, Fattaneh; Salehi, Mohammad; Bandehpour, Mojgan; Nariman, Mosaffa; Ghafari Novin, Marefat; Hosseini, Taher; Nematollahi, Sedigheh; Noroozian, Mohsen; Keshavarzi, Somayeh; Hosseini, Ahmad

    2014-01-01

    Nuclear transfer-embryonic stem cells (NT-ESCs) are genetically identical to the donor's cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A (TSA) as a histone deacetylase in- hibitor (HDACi) plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer (SCNT) can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines. In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6×DBA/2) F 1 mice] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were ac- tivated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts (MEF), so ESCs lines were estab- lished. ESCs markers were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Histone modifications were analyzed by enzyme linked immunosorbent assay (ELISA). Result of this study showed that TSA treatment after SCNT can improve devel- opmental rate of embryos (21.12 ± 3.56 vs. 8.08 ± 7.92), as well as establishment rate of NT-ESCs line (25 vs. 12.5). We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells (ESCs) lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs. TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a man- ner similar to that of ESCs established from normal blastocysts.

  11. Histone Modification of Embryonic Stem Cells Produced by Somatic Cell Nuclear Transfer and Fertilized Blastocysts

    PubMed Central

    Farifteh, Fattaneh; Salehi, Mohammad; Bandehpour, Mojgan; Nariman, Mosaffa; Ghafari Novin, Marefat; Hosseini, Taher; Nematollahi, Sedigheh; Noroozian, Mohsen; Keshavarzi, Somayeh; Hosseini, Ahmad

    2014-01-01

    Objective Nuclear transfer-embryonic stem cells (NT-ESCs) are genetically identical to the donor’s cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A (TSA) as a histone deacetylase in- hibitor (HDACi) plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer (SCNT) can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines. Materials and Methods In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6×DBA/2) F 1 mice] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were ac- tivated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts (MEF), so ESCs lines were estab- lished. ESCs markers were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Histone modifications were analyzed by enzyme linked immunosorbent assay (ELISA). Results Result of this study showed that TSA treatment after SCNT can improve devel- opmental rate of embryos (21.12 ± 3.56 vs. 8.08 ± 7.92), as well as establishment rate of NT-ESCs line (25 vs. 12.5). We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells (ESCs) lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs. Conclusion TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a man- ner similar to that of ESCs established from normal blastocysts. PMID:24381856

  12. Renal cell carcinoma–derived gangliosides suppress nuclear factor-κB activation in T cells

    PubMed Central

    Uzzo, Robert G.; Rayman, Patricia; Kolenko, Vladimir; Clark, Peter E.; Cathcart, Martha K.; Bloom, Tracy; Novick, Andrew C.; Bukowski, Ronald M.; Hamilton, Thomas; Finke, James H.

    1999-01-01

    Activation of the transcription factor nuclear factor-κB (NFκB) is impaired in T cells from patients with renal cell carcinomas (RCCs). In circulating T cells from a subset of patients with RCCs, the suppression of NFκB binding activity is downstream from the stimulus-induced degradation of the cytoplasmic factor IκBα. Tumor-derived soluble products from cultured RCC explants inhibit NFκB activity in T cells from healthy volunteers, despite a normal level of stimulus-induced IκBα degradation in these cells. The inhibitory agent has several features characteristic of a ganglioside, including sensitivity to neuraminidase but not protease treatment; hydrophobicity; and molecular weight less than 3 kDa. Indeed, we detected gangliosides in supernatants from RCC explants and not from adjacent normal kidney tissue. Gangliosides prepared from RCC supernatants, as well as the purified bovine gangliosides Gm1 and Gd1a, suppressed NFκB binding activity in T cells and reduced expression of the cytokines IL-2 and IFN-γ. Taken together, our findings suggest that tumor-derived gangliosides may blunt antitumor immune responses in patients with RCCs. PMID:10491412

  13. Epigenetic reprogramming by somatic cell nuclear transfer: questions and potential solutions.

    PubMed

    Huili, Ji; Haosheng, Lu; Dengke, Pan

    2014-12-01

    Somatic cell nuclear transfer (SCNT) is a technology by which a highly differentiated somatic nucleus is transferred into an enucleated oocyte to generate a reconstructed embryo that subsequently develops to an offspring. However, to date, the efficiency of cloned animal is still low. The major reason is incomplete nuclear reprogramming of donor cells after nuclear transfer, which results in abnormal epigenetic modifications, including DNA methylation, histone acetylation, gene imprinting, X-chromosome inactivation, and telomere length. Most improvements have been made in somatic epigenetic reprogramming with small molecules and manipulating expression of specific genes. It is expected that SCNT will soon have broad applications in both basic research and practical production. In this review, we summarize the recent progress in epigenetic reprogramming by somatic cell nuclear transfer; in particular, we focus on strategies for rescuing the epigenetic errors occurring during SCNT.

  14. Cell cycle-dependent alteration in NAC1 nuclear body dynamics and morphology

    NASA Astrophysics Data System (ADS)

    Wu, Pei-Hsun; Hung, Shen-Hsiu; Ren, Tina; Shih, Ie-Ming; Tseng, Yiider

    2011-02-01

    NAC1, a BTB/POZ family member, has been suggested to participate in maintaining the stemness of embryonic stem cells and has been implicated in the pathogenesis of human cancer. In ovarian cancer, NAC1 upregulation is associated with disease aggressiveness and with the development of chemoresistance. Like other BTB/POZ proteins, NAC1 forms discrete nuclear bodies in non-dividing cells. To investigate the biological role of NAC1 nuclear bodies, we characterized the expression dynamics of NAC1 nuclear bodies during different phases of the cell cycle. Fluorescence recovery after photobleaching assays revealed that NAC1 was rapidly exchanged between the nucleoplasm and NAC1 nuclear bodies in interphase cells. The number of NAC1 bodies significantly increased and their size decreased in the S phase as compared to the G0/G1 and G2 phases. NAC1 nuclear bodies disappeared and NAC1 became diffuse during mitosis. NAC1 nuclear bodies reappeared immediately after completion of mitosis. These results indicate that a cell cycle-dependent regulatory mechanism controls NAC1 body formation in the nucleus and suggest that NAC1 body dynamics are associated with mitosis or cytokinesis.

  15. Granulocytic nuclear differentiation of lamin B receptor-deficient mouse EPRO cells.

    PubMed

    Zwerger, Monika; Herrmann, Harald; Gaines, Peter; Olins, Ada L; Olins, Donald E

    2008-08-01

    Lamin B receptor (LBR) is an integral protein of the inner nuclear membrane. Recent studies have demonstrated that genetic deficiency of LBR during granulopoiesis results in hypolobulation of the mature neutrophil nucleus, as observed in human Pelger-Huët anomaly and mouse ichthyosis (ic). In this study, we utilized differentiated early promyelocytes (EPRO cells) that were derived from the bone marrow of homozygous and heterozygous ichthyosis mice to examine changes to the expression of nuclear envelope proteins and heterochromatin structure that result from deficient LBR expression. Wild-type (+/+), heterozygous (+/ic), and homozygous (ic/ic) granulocytic forms of EPRO cells were analyzed for the expression of multiple lamins and inner nuclear envelope proteins by immunostaining and immunoblotting techniques. The heterochromatin architecture was also examined by immunostaining for histone lysine methylation. Wild-type (+/+) and heterozygous (+/ic) granulocytic forms revealed ring-shaped nuclei and contained LBR within the nuclear envelope; ic/ic granulocytes exhibited smaller ovoid nuclei devoid of LBR. The pericentric heterochromatin of undifferentiated and granulocytic ic/ic cells was condensed into larger spots and shifted away from the nuclear envelope, compared to +/+ and +/ic cell forms. Lamin A/C, which is normally not present in mature granulocytes, was significantly elevated in LBR-deficient EPRO cells. Our observations suggest roles for LBR during granulopoiesis, which can involve augmenting nuclear membrane growth, facilitating compartmentalization of heterochromatin, and promoting downregulation of lamin A/C expression.

  16. In silico synchronization reveals regulators of nuclear ruptures in lamin A/C deficient model cells

    PubMed Central

    Robijns, J.; Molenberghs, F.; Sieprath, T.; Corne, T. D. J.; Verschuuren, M.; De Vos, W. H.

    2016-01-01

    The nuclear lamina is a critical regulator of nuclear structure and function. Nuclei from laminopathy patient cells experience repetitive disruptions of the nuclear envelope, causing transient intermingling of nuclear and cytoplasmic components. The exact causes and consequences of these events are not fully understood, but their stochastic occurrence complicates in-depth analyses. To resolve this, we have established a method that enables quantitative investigation of spontaneous nuclear ruptures, based on co-expression of a firmly bound nuclear reference marker and a fluorescent protein that shuttles between the nucleus and cytoplasm during ruptures. Minimally invasive imaging of both reporters, combined with automated tracking and in silico synchronization of individual rupture events, allowed extracting information on rupture frequency and recovery kinetics. Using this approach, we found that rupture frequency correlates inversely with lamin A/C levels, and can be reduced in genome-edited LMNA knockout cells by blocking actomyosin contractility or inhibiting the acetyl-transferase protein NAT10. Nuclear signal recovery followed a kinetic that is co-determined by the severity of the rupture event, and could be prolonged by knockdown of the ESCRT-III complex component CHMP4B. In conclusion, our approach reveals regulators of nuclear rupture induction and repair, which may have critical roles in disease development. PMID:27461848

  17. In silico synchronization reveals regulators of nuclear ruptures in lamin A/C deficient model cells

    NASA Astrophysics Data System (ADS)

    Robijns, J.; Molenberghs, F.; Sieprath, T.; Corne, T. D. J.; Verschuuren, M.; de Vos, W. H.

    2016-07-01

    The nuclear lamina is a critical regulator of nuclear structure and function. Nuclei from laminopathy patient cells experience repetitive disruptions of the nuclear envelope, causing transient intermingling of nuclear and cytoplasmic components. The exact causes and consequences of these events are not fully understood, but their stochastic occurrence complicates in-depth analyses. To resolve this, we have established a method that enables quantitative investigation of spontaneous nuclear ruptures, based on co-expression of a firmly bound nuclear reference marker and a fluorescent protein that shuttles between the nucleus and cytoplasm during ruptures. Minimally invasive imaging of both reporters, combined with automated tracking and in silico synchronization of individual rupture events, allowed extracting information on rupture frequency and recovery kinetics. Using this approach, we found that rupture frequency correlates inversely with lamin A/C levels, and can be reduced in genome-edited LMNA knockout cells by blocking actomyosin contractility or inhibiting the acetyl-transferase protein NAT10. Nuclear signal recovery followed a kinetic that is co-determined by the severity of the rupture event, and could be prolonged by knockdown of the ESCRT-III complex component CHMP4B. In conclusion, our approach reveals regulators of nuclear rupture induction and repair, which may have critical roles in disease development.

  18. Identification and Ultrastructural Characterization of a Novel Nuclear Degradation Complex in Differentiating Lens Fiber Cells

    PubMed Central

    Costello, M. Joseph; Brennan, Lisa A.; Gilliland, Kurt O.; Johnsen, Sönke; Kantorow, Marc

    2016-01-01

    An unresolved issue in structural biology is how the encapsulated lens removes membranous organelles to carry out its role as a transparent optical element. In this ultrastructural study, we establish a mechanism for nuclear elimination in the developing chick lens during the formation of the organelle-free zone. Day 12–15 chick embryo lenses were examined by high-resolution confocal light microscopy and thin section transmission electron microscopy (TEM) following fixation in 10% formalin and 4% paraformaldehyde, and then processing for confocal or TEM as described previously. Examination of developing fiber cells revealed normal nuclei with dispersed chromatin and clear nucleoli typical of cells in active ribosome production to support protein synthesis. Early signs of nuclear degradation were observed about 300 μm from the lens capsule in Day 15 lenses where the nuclei display irregular nuclear stain and prominent indentations that sometimes contained a previously undescribed macromolecular aggregate attached to the nuclear envelope. We have termed this novel structure the nuclear excisosome. This complex by confocal is closely adherent to the nuclear envelope and by TEM appears to degrade the outer leaflet of the nuclear envelope, then the inner leaflet up to 500 μm depth. The images suggest that the nuclear excisosome separates nuclear membrane proteins from lipids, which then form multilamellar assemblies that stain intensely in confocal and in TEM have 5 nm spacing consistent with pure lipid bilayers. The denuded nucleoplasm then degrades by condensation and loss of structure in the range 600 to 700 μm depth producing pyknotic nuclear remnants. None of these stages display any classic autophagic vesicles or lysosomes associated with nuclei. Uniquely, the origin of the nuclear excisosome is from filopodial-like projections of adjacent lens fiber cells that initially contact, and then appear to fuse with the outer nuclear membrane. These filopodial

  19. Alterations in the nuclear proteome of HIV-1 infected T-cells

    SciTech Connect

    DeBoer, Jason; Jagadish, Teena; Haverland, Nicole A.; Madson, Christian J.; Ciborowski, Pawel; Belshan, Michael

    2014-11-15

    Virus infection of a cell involves the appropriation of host factors and the innate defensive response of the cell. The identification of proteins critical for virus replication may lead to the development of novel, cell-based inhibitors. In this study we mapped the changes in T-cell nuclei during human immunodeficiency virus type 1 (HIV-1) at 20 hpi. Using a stringent data threshold, a total of 13 and 38 unique proteins were identified in infected and uninfected cells, respectively, across all biological replicates. An additional 15 proteins were found to be differentially regulated between infected and control nuclei. STRING analysis identified four clusters of protein–protein interactions in the data set related to nuclear architecture, RNA regulation, cell division, and cell homeostasis. Immunoblot analysis confirmed the differential expression of several proteins in both C8166-45 and Jurkat E6-1 T-cells. These data provide a map of the response in host cell nuclei upon HIV-1 infection. - Highlights: • We identify changes in the expression of nuclear proteins during HIV-1 infection. • 163 nuclear proteins were found differentially regulated during HIV-1 infection. • Bioinformatic analysis identified several nuclear pathways altered by HIV infection. • Candidate factors were validated in two independent cell lines.

  20. Nuclear Membranes ETB Receptors Mediate ET-1-induced Increase of Nuclear Calcium in Human Left Ventricular Endocardial Endothelial Cells.

    PubMed

    Jules, Farah; Avedanian, Levon; Al-Khoury, Johny; Keita, Ramatoulaye; Normand, Alexandre; Bkaily, Ghassan; Jacques, Danielle

    2015-07-01

    In fetal human left ventricular endocardial endothelial cells (EECLs), both plasma membrane (PM) ET(A)R and ET(B)R were reported to mediate ET-1-induced increase of intracellular calcium [Ca](i); however, this effect was mediated by ET(A)R in right EECs (EECRs). In this study, we verified whether, as for the PM, nuclear membranes (NMs) ET-1 receptors activation in EECLs and EECRs induce an increase of nuclear calcium ([Ca](n)) and if this effect is mediated through the same receptor type as in PM. Using a plasmalemma-perforated technique and 3D confocal microscopy, our results showed that, as in PM intact cells, superfusion of nuclei of both cell types with cytosolic ET-1 induced a concentration-dependent sustained increase of [Ca](n). In EECRs, the ET(A)R antagonist prevented the effect of ET-1 on [Ca](n) without affecting EECLs. However, in both cell types, the effect of cytosolic ET-1 on [Ca](n) was prevented by the ETBR antagonist. In conclusion, both NMs' ET(A)R and ET(B)R mediated the effect of cytosolic ET-1 on [Ca](n) in EECRs. In contrast, only NMs' ET(B)R activation mediated the effect of cytosolic ET-1 in EECLs. Hence, the type of NMs' receptors mediating the effect of ET-1 on [Ca](n) are different from those of PM mediating the increase in [Ca](i).

  1. Cell-mediated transgenesis in rabbits: chimeric and nuclear transfer animals.

    PubMed

    Zakhartchenko, V; Flisikowska, T; Li, S; Richter, T; Wieland, H; Durkovic, M; Rottmann, O; Kessler, B; Gungor, T; Brem, G; Kind, A; Wolf, E; Schnieke, A

    2011-02-01

    The ability to perform precise genetic engineering such as gene targeting in rabbits would benefit biomedical research by enabling, for example, the generation of genetically defined rabbit models of human diseases. This has so far not been possible because of the lack of functional rabbit embryonic stem cells and the high fetal and perinatal mortality associated with rabbit somatic cell nuclear transfer. We examined cultured pluripotent and multipotent cells for their ability to support the production of viable animals. Rabbit putative embryonic stem (ES) cells were derived and shown capable of in vitro and in vivo pluripotent differentiation. We report the first live born ES-derived rabbit chimera. Rabbit mesenchymal stem cells (MSCs) were derived from bone marrow, and multipotent differentiation was demonstrated in vitro. Nuclear transfer was carried out with both cell types, and embryo development was assessed in vitro and in vivo. Rabbit MSCs were markedly more successful than ES cells as nuclear donors. MSCs were transfected with fluorescent reporter gene constructs and assessed for nuclear transfer competence. Transfected MSCs supported development with similar efficiency as normal MSCs and resulted in the first live cloned rabbits from genetically manipulated MSCs. Reactivation of fluorescence reporter gene expression in reconstructed embryos was investigated as a means of identifying viable embryos in vitro but was not a reliable predictor. We also examined serial nuclear transfer as a means of rescuing dead animals.

  2. Acoustic sensors for fission gas characterization: R and D skills devoted to innovative instrumentation in MTR, non-destructive devices in hot lab facilities and specific transducers for measurements of LWR rods in nuclear plants

    SciTech Connect

    Ferrandis, J.Y.; Leveque, G.; Rosenkrantz, E.; Augereau, F.; Combette, P.

    2015-07-01

    irradiation. The instrumented fuel rod incorporating the ultrasonic gas composition sensor was finally irradiated during 2 weeks in nominal conditions. Neutronics calculation will be performed in order to calculate the thermal and fast neutron fluence and the gamma dose absorbed by acoustic sensor. A first evaluation gives a thermal fluence about 4,5.10{sup 19} n/cm{sup 2}, a fast neutrons fluence about 4,5.1018 n/cm{sup 2} and a total gamma dose up to 0,25 MGy The maximal temperature during the irradiation test was about 150 C. Although the ultrasonic sensor appears to be damaged, the optimization of the electrical attack parameters and the development of a new signal processing maintain the measurement feasibility up the end of the irradiation campaign. It was the first time that the composition of fission gas has been monitored all along an irradiation experiment in a MTR, giving access to the gas release kinetics. New researches involve thick film transducers produced by screen-printing process in order to propose piezoelectric structures for harsh temperature and irradiation measurements. The second project consists in the development of a non-destructive device that can be directly applied on a LWR fuel rod. The problem to be solved relates to the measurement of the fission gas pressure and composition in a fuel rod using a non-destructive method. Fuel rod internal pressure is one of the safety criteria applied in nuclear power analyses. This criterion must be verified in order to avoid any fuel-cladding gap reopening risk and therefore any local clad ballooning. Apart from the safety implications, this parameter is also a fuel behaviour indicator and reflects the overall fuel performance in operation, but also during shipping and long-term storage. Rod internal pressure is one criterion amongst others, like cladding corrosion, against which the acceptable fuel burn-up limit is set. A sensor has been achieved in 2007. A full-scale hot cell test of the internal gas

  3. Nuclear factor of activated T cells (NFAT) signaling regulates PTEN expression and intestinal cell differentiation

    PubMed Central

    Wang, Qingding; Zhou, Yuning; Jackson, Lindsey N.; Johnson, Sara M.; Chow, Chi-Wing; Evers, B. Mark

    2011-01-01

    The nuclear factor of activated T cell (NFAT) proteins are a family of transcription factors (NFATc1–c4) involved in the regulation of cell differentiation and adaptation. Previously we demonstrated that inhibition of phosphatidylinositol 3-kinase or overexpression of PTEN enhanced intestinal cell differentiation. Here we show that treatment of intestinal-derived cells with the differentiating agent sodium butyrate (NaBT) increased PTEN expression, NFAT binding activity, and NFAT mRNA expression, whereas pretreatment with the NFAT signaling inhibitor cyclosporine A (CsA) blocked NaBT-mediated PTEN induction. Moreover, knockdown of NFATc1 or NFATc4, but not NFATc2 or NFATc3, attenuated NaBT-induced PTEN expression. Knockdown of NFATc1 decreased PTEN expression and increased the phosphorylation levels of Akt and downstream targets Foxo1 and GSK-3α/β. Furthermore, overexpression of NFATc1 or the NFATc4 active mutant increased PTEN and p27kip1 expression and decreased Akt phosphorylation. In addition, pretreatment with CsA blocked NaBT-mediated induction of intestinal alkaline phosphatase (IAP) activity and villin and p27kip1 expression; knockdown of either NFATc1 or NFATc4 attenuated NaBT-induced IAP activity. We provide evidence showing that NFATc1 and NFATc4 are regulators of PTEN expression. Importantly, our results suggest that NFATc1 and NFATc4 regulation of intestinal cell differentiation may be through PTEN regulation. PMID:21148296

  4. Angiomotin promotes renal epithelial and carcinoma cell proliferation by retaining the nuclear YAP.

    PubMed

    Lv, Meng; Li, Shuting; Luo, Changqin; Zhang, Xiaoman; Shen, Yanwei; Sui, Yan Xia; Wang, Fan; Wang, Xin; Yang, Jiao; Liu, Peijun; Yang, Jin

    2016-03-15

    Renal cell carcinoma (RCC) is one of the common tumors in the urinary system without effective therapies. Angiomotin (Amot) can interact with Yes-associated protein (YAP) to either stimulate or inhibit YAP activity, playing a potential role in cell proliferation. However, the role of Amot in regulating the proliferation of renal epithelial and RCC cells is unknown. Here, we show that Amot is expressed predominantly in the nucleus of RCC cells and tissues, and in the cytoplasm and nucleus of renal epithelial cells and paracancerous tissues. Furthermore, Amot silencing inhibited proliferation of HK-2 and 786-O cells while Amot upregulation promoted proliferation of ACHN cells. Interestingly, the location of Amot and YAP in RCC clinical samples and cells was similar. Amot interacted with YAP in HK-2 and 786-O cells, particularly in the nucleus. Moreover, Amot silencing mitigated the levels of nuclear YAP in HK-2 and 786-O cells and reduced YAP-related CTGF and Cyr61 expression in 786-O cells. Amot upregulation slightly increased the nuclear YAP and YAP-related gene expression in ACHN cells. Finally, enhanced YAP expression restored proliferation of Amot-silencing 786-O cells. Together, these data indicate that Amot is crucial for the maintenance of nuclear YAP to promote renal epithelial and RCC proliferation.

  5. Murine leukemia virus vector integration favors promoter regions and regional hot spots in a human T-cell line

    SciTech Connect

    Tsukahara, Tomonori; Agawa, Hideyuki; Matsumoto, Sayori; Matsuda, Mizuho; Ueno, Shuichi; Yamashita, Yuki; Yamada, Koichiro; Tanaka, Nobuyuki; Kojima, Katsuhiko; Takeshita, Toshikazu . E-mail: takesit@sch.md.shinshu-u.ac.jp

    2006-07-07

    Genomic analysis of integration will be important in evaluating the safety of human gene therapy with retroviral vectors. Here, we investigated MLV vector integration sites in human T-cells, since they are amenable to gene transfer studies, and have been used therapeutically in clinical trials. We mapped 340 MLV vector integration sites in the infected human T-cell clones we established. The data showed that MLV preferred integration near the transcription start sites ({+-}5 kb), near CpG islands ({+-}1 kb), and within the first intron of RefSeq genes. We also identified MLV integration hot spots that contained three or more integrations within a 100 kb region. RT-PCR revealed that mRNA-levels of T-cell clones that contained MLV integrations near transcription start sites or introns were dysregulated compared to the uninfected cells. These studies help define the profile of MLV integration in T-cells and the risks associated with MLV-based gene therapy.

  6. Hot Groups.

    ERIC Educational Resources Information Center

    Vail, Kathleen

    1996-01-01

    Collaborators sparked by creative ideas and obsessed by a common task may not realize they're part of a "hot group"--a term coined by business professors Harold J. Leavitt and Jean Lipman-Blumen. Spawned by group decision making and employee empowerment, hot groups can flourish in education settings. They're typically small, short lived,…

  7. Hot-electron luminescence in aged electrodeposited CdSe liquid-junction solar cell

    SciTech Connect

    Silberstein, R.P.; Tomkiewicz, M.

    1983-01-01

    We have utilized Raman spectroscopy and scanning Auger electron spectroscopy (AES) to probe the surface of polycrystalline, electrodeposited CdSe photoelectrodes which have been aged in a polysulfide electrolyte under illumination and in darkness. We have observed characteristic ''hot-electron'' luminescence at multiples of ..omega../sub LO/ (CdS) = 305 cm/sup -1/ in the light-aged electrode, indicating that a surface layer of CdS has been formed. AES profiling shows that extensive substitution of S for Se has occurred, in the light-aged electrode alone, to a depth of approx.600 A. Measurements at 300 K suggest that Raman scattering can be a useful, in situ, contactless, nondestructive probe of CdS formation.

  8. High throughput sequencing identifies an imprinted gene, Grb10, associated with the pluripotency state in nuclear transfer embryonic stem cells.

    PubMed

    Li, Hui; Gao, Shuai; Huang, Hua; Liu, Wenqiang; Huang, Huanwei; Liu, Xiaoyu; Gao, Yawei; Le, Rongrong; Kou, Xiaochen; Zhao, Yanhong; Kou, Zhaohui; Li, Jia; Wang, Hong; Zhang, Yu; Wang, Hailin; Cai, Tao; Sun, Qingyuan; Gao, Shaorong; Han, Zhiming

    2017-07-18

    Somatic cell nuclear transfer and transcription factor mediated reprogramming are two widely used techniques for somatic cell reprogramming. Both fully reprogrammed nuclear transfer embryonic stem cells and induced pluripotent stem cells hold potential for regenerative medicine, and evaluation of the stem cell pluripotency state is crucial for these applications. Previous reports have shown that the Dlk1-Dio3 region is associated with pluripotency in induced pluripotent stem cells and the incomplete somatic cell reprogramming causes abnormally elevated levels of genomic 5-methylcytosine in induced pluripotent stem cells compared to nuclear transfer embryonic stem cells and embryonic stem cells. In this study, we compared pluripotency associated genes Rian and Gtl2 in the Dlk1-Dio3 region in exactly syngeneic nuclear transfer embryonic stem cells and induced pluripotent stem cells with same genomic insertion. We also assessed 5-methylcytosine and 5-hydroxymethylcytosine levels and performed high-throughput sequencing in these cells. Our results showed that Rian and Gtl2 in the Dlk1-Dio3 region related to pluripotency in induced pluripotent stem cells did not correlate with the genes in nuclear transfer embryonic stem cells, and no significant difference in 5-methylcytosine and 5-hydroxymethylcytosine levels were observed between fully and partially reprogrammed nuclear transfer embryonic stem cells and induced pluripotent stem cells. Through syngeneic comparison, our study identifies for the first time that Grb10 is associated with the pluripotency state in nuclear transfer embryonic stem cells.

  9. Changing nuclear landscape and unique PML structures during early epigenetic transitions of human embryonic stem cells.

    PubMed

    Butler, John T; Hall, Lisa L; Smith, Kelly P; Lawrence, Jeanne B

    2009-07-01

    The complex nuclear structure of somatic cells is important to epigenomic regulation, yet little is known about nuclear organization of human embryonic stem cells (hESC). Here we surveyed several nuclear structures in pluripotent and transitioning hESC. Observations of centromeres, telomeres, SC35 speckles, Cajal Bodies, lamin A/C and emerin, nuclear shape and size demonstrate a very different "nuclear landscape" in hESC. This landscape is remodeled during a brief transitional window, concomitant with or just prior to differentiation onset. Notably, hESC initially contain abundant signal for spliceosome assembly factor, SC35, but lack discrete SC35 domains; these form as cells begin to specialize, likely reflecting cell-type specific genomic organization. Concomitantly, nuclear size increases and shape changes as lamin A/C and emerin incorporate into the lamina. During this brief window, hESC exhibit dramatically different PML-defined structures, which in somatic cells are linked to gene regulation and cancer. Unlike the numerous, spherical somatic PML bodies, hES cells often display approximately 1-3 large PML structures of two morphological types: long linear "rods" or elaborate "rosettes", which lack substantial SUMO-1, Daxx, and Sp100. These occur primarily between Day 0-2 of differentiation and become rare thereafter. PML rods may be "taut" between other structures, such as centromeres, but clearly show some relationship with the lamina, where PML often abuts or fills a "gap" in early lamin A/C staining. Findings demonstrate that pluripotent hES cells have a markedly different overall nuclear architecture, remodeling of which is linked to early epigenomic programming and involves formation of unique PML-defined structures.

  10. Fission yeast cells undergo nuclear division in the absence of spindle microtubules.

    PubMed

    Castagnetti, Stefania; Oliferenko, Snezhana; Nurse, Paul

    2010-10-12

    Mitosis in eukaryotic cells employs spindle microtubules to drive accurate chromosome segregation at cell division. Cells lacking spindle microtubules arrest in mitosis due to a spindle checkpoint that delays mitotic progression until all chromosomes have achieved stable bipolar attachment to spindle microtubules. In fission yeast, mitosis occurs within an intact nuclear membrane with the mitotic spindle elongating between the spindle pole bodies. We show here that in fission yeast interference with mitotic spindle formation delays mitosis only briefly and cells proceed to an unusual nuclear division process we term nuclear fission, during which cells perform some chromosome segregation and efficiently enter S-phase of the next cell cycle. Nuclear fission is blocked if spindle pole body maturation or sister chromatid separation cannot take place or if actin polymerization is inhibited. We suggest that this process exhibits vestiges of a primitive nuclear division process independent of spindle microtubules, possibly reflecting an evolutionary intermediate state between bacterial and Archeal chromosome segregation where the nucleoid divides without a spindle and a microtubule spindle-based eukaryotic mitosis.

  11. Fission Yeast Cells Undergo Nuclear Division in the Absence of Spindle Microtubules

    PubMed Central

    Castagnetti, Stefania; Oliferenko, Snezhana; Nurse, Paul

    2010-01-01

    Mitosis in eukaryotic cells employs spindle microtubules to drive accurate chromosome segregation at cell division. Cells lacking spindle microtubules arrest in mitosis due to a spindle checkpoint that delays mitotic progression until all chromosomes have achieved stable bipolar attachment to spindle microtubules. In fission yeast, mitosis occurs within an intact nuclear membrane with the mitotic spindle elongating between the spindle pole bodies. We show here that in fission yeast interference with mitotic spindle formation delays mitosis only briefly and cells proceed to an unusual nuclear division process we term nuclear fission, during which cells perform some chromosome segregation and efficiently enter S-phase of the next cell cycle. Nuclear fission is blocked if spindle pole body maturation or sister chromatid separation cannot take place or if actin polymerization is inhibited. We suggest that this process exhibits vestiges of a primitive nuclear division process independent of spindle microtubules, possibly reflecting an evolutionary intermediate state between bacterial and Archeal chromosome segregation where the nucleoid divides without a spindle and a microtubule spindle-based eukaryotic mitosis. PMID:20967237

  12. RIMA-dependent nuclear accumulation of IYO triggers auxin-irreversible cell differentiation in Arabidopsis.

    PubMed

    Muñoz, Alfonso; Mangano, Silvina; González-García, Mary Paz; Contreras, Ramón; Sauer, Michael B; De Rybel, Bert; Weijers, Dolf; Sánchez-Serrano, José J; Sanmartín, Maite; Rojo, Enrique

    2017-02-21

    The transcriptional regulator MINIYO (IYO) is essential and rate-limiting for initiating cell differentiation in Arabidopsis thaliana. Moreover, IYO moves from the cytosol into the nucleus in cells at the meristem periphery, possibly triggering their differentiation. However, the genetic mechanisms controlling IYO nuclear accumulation were unknown and the evidence that increased nuclear IYO levels trigger differentiation remained correlative. Searching for IYO interactors, we have identified RPAP2 IYO Mate (RIMA), a homologue of yeast and human proteins linked to nuclear import of selective cargo. Knockdown of RIMA causes delayed onset of cell differentiation, phenocopying the effects of IYO knock down at the transcriptomic and developmental levels. Moreover, differentiation is completely blocked when IYO and RIMA activities are simultaneously reduced and is synergistically accelerated when IYO and RIMA are concurrently overexpressed, confirming their functional interaction. Indeed, RIMA knockdown reduces the nuclear levels of IYO and prevents its pro-differentiation activity, supporting the conclusion that RIMA-dependent nuclear IYO accumulation triggers cell differentiation in Arabidopsis. Importantly, by analysing the effect of the IYO/RIMA pathway on xylem pole pericycle cells, we provide compelling evidence reinforcing the view that the capacity for de novo organogenesis and regeneration from mature plant tissues can reside in stem cell reservoirs.

  13. Mechano-topographic modulation of stem cell nuclear shape on nanofibrous scaffolds.

    PubMed

    Nathan, Ashwin S; Baker, Brendon M; Nerurkar, Nandan L; Mauck, Robert L

    2011-01-01

    Stem cells transit along a variety of lineage-specific routes towards differentiated phenotypes. These fate decisions are dependent not just on the soluble chemical cues that are encountered or enforced in vivo and in vitro, but also on physical cues from the cellular microenvironment. These physical cues can consist of both nano- and micro-scale topographical features, as well as mechanical inputs provided passively (from the base properties of the materials to which they adhere) or actively (from extrinsic applied mechanical deformations). A suitable tool to investigate the coordination of these cues lies in nanofibrous scaffolds, which can both dictate cellular and cytoskeletal orientation and facilitate mechanical perturbation of seeded cells. Here, we demonstrate a coordinated influence of scaffold architecture (aligned vs. randomly organized fibers) and tensile deformation on nuclear shape and orientation. Sensitivity of nuclear morphology to scaffold architecture was more pronounced in stem cell populations than in terminally differentiated fibrochondrocytes. Tension applied to the scaffold elicited further alterations in nuclear morphology, greatest in stem cells, that were mediated by the filamentous actin cytoskeleton, but not the microtubule or intermediate filament network. Nuclear perturbations were time and direction dependent, suggesting that the modality and direction of loading influenced nuclear architecture. The present work may provide additional insight into the mechanisms by which the physical microenvironment influences cell fate decisions, and has specific application to the design of new materials for regenerative medicine applications with adult stem cells. Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  14. Visualizing the molecular sociology at the HeLa cell nuclear periphery.

    PubMed

    Mahamid, Julia; Pfeffer, Stefan; Schaffer, Miroslava; Villa, Elizabeth; Danev, Radostin; Cuellar, Luis Kuhn; Förster, Friedrich; Hyman, Anthony A; Plitzko, Jürgen M; Baumeister, Wolfgang

    2016-02-26

    The molecular organization of eukaryotic nuclear volumes remains largely unexplored. Here we combined recent developments in cryo-electron tomography (cryo-ET) to produce three-dimensional snapshots of the HeLa cell nuclear periphery. Subtomogram averaging and classification of ribosomes revealed the native structure and organization of the cytoplasmic translation machinery. Analysis of a large dynamic structure-the nuclear pore complex-revealed variations detectable at the level of individual complexes. Cryo-ET was used to visualize previously elusive structures, such as nucleosome chains and the filaments of the nuclear lamina, in situ. Elucidation of the lamina structure provides insight into its contribution to metazoan nuclear stiffness. Copyright © 2016, American Association for the Advancement of Science.

  15. Nuclear reprogramming: the strategy used in normal development is also used in somatic cell nuclear transfer and parthenogenesis.

    PubMed

    Gao, Tianlong; Zheng, Junke; Xing, Fengying; Fang, Haiyan; Sun, Feng; Yan, Ayong; Gong, Xun; Ding, Hui; Tang, Fan; Sheng, Hui Z

    2007-02-01

    Somatic cell nuclear transfer (SCNT) and parthenogenesis are alternative forms of reproduction and development, building new life cycles on differentiated somatic cell nuclei and duplicated maternal chromatin, respectively. In the preceding paper (Sun F, et al., Cell Res 2007; 17:117-134.), we showed that an "erase-and-rebuild" strategy is used in normal development to transform the maternal gene expression profile to a zygotic one. Here, we investigate if the same strategy also applies to SCNT and parthenogenesis. The relationship between chromatin and chromatin factors (CFs) during SCNT and parthenogenesis was examined using immunochemical and GFP-fusion protein assays. Results from these studies indicated that soon after nuclear transfer, a majority of CFs dissociated from somatic nuclei and were redistributed to the cytoplasm of the egg. The erasure process in oogenesis is recaptured during the initial phase in SCNT. Most CFs entered pseudo-pronuclei shortly after their formation. In parthenogenesis, all parthenogenotes underwent normal oogenesis, and thus had removed most CFs from chromosomes before the initiation of development. The CFs were subsequently re-associated with female pronuclei in time and sequence similar to that in fertilized embryos. Based on these data, we conclude that the "erase-and-rebuild" process observed in normal development also occurs in SCNT and in parthenogenesis, albeit in altered fashions. The process is responsible for transcription reprogramming in these procedures. The "erase" process in SCNT is compressed and the efficiency is compromised, which likely contribute to the developmental defects often observed in nuclear transfer (nt) embryos. Furthermore, results from this study indicated that the cytoplasm of an egg contains most, if not all, essential components for assembling the zygotic program and can assemble them onto appropriate diploid chromatin of distinct origins.

  16. Proliferating cell nuclear antigen (PCNA): a key factor in DNA replication and cell cycle regulation.

    PubMed

    Strzalka, Wojciech; Ziemienowicz, Alicja

    2011-05-01

    PCNA (proliferating cell nuclear antigen) has been found in the nuclei of yeast, plant and animal cells that undergo cell division, suggesting a function in cell cycle regulation and/or DNA replication. It subsequently became clear that PCNA also played a role in other processes involving the cell genome. This review discusses eukaryotic PCNA, with an emphasis on plant PCNA, in terms of the protein structure and its biochemical properties as well as gene structure, organization, expression and function. PCNA exerts a tripartite function by operating as (1) a sliding clamp during DNA synthesis, (2) a polymerase switch factor and (3) a recruitment factor. Most of its functions are mediated by its interactions with various proteins involved in DNA synthesis, repair and recombination as well as in regulation of the cell cycle and chromatid cohesion. Moreover, post-translational modifications of PCNA play a key role in regulation of its functions. Finally, a phylogenetic comparison of PCNA genes suggests that the multi-functionality observed in most species is a product of evolution. Most plant PCNAs exhibit features similar to those found for PCNAs of other eukaryotes. Similarities include: (1) a trimeric ring structure of the PCNA sliding clamp, (2) the involvement of PCNA in DNA replication and repair, (3) the ability to stimulate the activity of DNA polymerase δ and (4) the ability to interact with p21, a regulator of the cell cycle. However, many plant genomes seem to contain the second, probably functional, copy of the PCNA gene, in contrast to PCNA pseudogenes that are found in mammalian genomes.

  17. Reduction of nuclear encoded enzymes of mitochondrial energy metabolism in cells devoid of mitochondrial DNA.

    PubMed

    Mueller, Edith E; Mayr, Johannes A; Zimmermann, Franz A; Feichtinger, René G; Stanger, Olaf; Sperl, Wolfgang; Kofler, Barbara

    2012-01-20

    Mitochondrial DNA (mtDNA) depletion syndromes are generally associated with reduced activities of oxidative phosphorylation (OXPHOS) enzymes that contain subunits encoded by mtDNA. Conversely, entirely nuclear encoded mitochondrial enzymes in these syndromes, such as the tricarboxylic acid cycle enzyme citrate synthase (CS) and OXPHOS complex II, usually exhibit normal or compensatory enhanced activities. Here we report that a human cell line devoid of mtDNA (HEK293 ρ(0) cells) has diminished activities of both complex II and CS. This finding indicates the existence of a feedback mechanism in ρ(0) cells that downregulates the expression of entirely nuclear encoded components of mitochondrial energy metabolism.

  18. Response of sheep lymphocytes to PHA: quantitation by nuclear volume measurement and cell counts (40764)

    SciTech Connect

    Chandra, P.; Chanana, A.D.; Joel, D.D.

    1980-03-01

    Phytohemagglutinin response of peripheral blood lymphocytes (PBL) of sheep was studied. Assessment of proliferative response was performed by determination of nuclear volumes and cell counts in cultures from 14 sheep and by incorporation of tritiated thymidine in cultures in four additional sheep. PBL of sheep were found to transform and proliferate with PHA similarly to human peripheral blood lymphocytes with minor differences. Quantitation of the proliferative response by determining the cell count and nuclear volumes provided more information on cell kinetics in culture than the commonly used isotope-labeled thymidine incorporation method.

  19. Nuclear size as a cell-kinetic marker for osteoblast differentiation

    NASA Technical Reports Server (NTRS)

    Roberts, W. E.; Mozsary, P. G.; Klingler, E.

    1982-01-01

    A nuclear morphometric assay for preosteoblasts is introduced as a cell-kinetic technique, applicable to routine histological preparations of mineralized tissue. Because this method is a morphological marker for osteoblast precursor cell differentiation, it provides a new dimension for determining the mechanism of osteoblast histogenesis. Osteoblast precursors of the periodontal ligament are a mixed population of progenitors, kinetically separable into two distinct groups according to nuclear size. Preosteoblasts, the immediate proliferating precursors of osteoblasts, have large nuclei (greater than 170 micrometers3) and are derived from relatively undifferentiated fibroblastlike cells, which have smaller nuclei (less than 80 micrometers3). Increase in nuclear volume, during G1 phase of the cell cycle, is apparently a morphological manifestation of change in genomic expression. This key event in preosteoblast differentiation is related to mechanical stress/strain and may be an important rate-limiting step in osteoblast histogenesis.

  20. Fludarabine nucleoside modulates nuclear "survival and death" proteins in resistant chronic lymphocytic leukemia cells.

    PubMed

    Henrich, Silke; Mactier, Swetlana; Best, Giles; Mulligan, Stephen P; Crossett, Ben; Christopherson, Richard Ian

    2011-12-01

    The nuclear mechanisms by which fludarabine nucleoside (F-ara-A) induces apoptosis have been investigated in human MEC1 cells derived from B-cell chronic lymphocytic leukemia. Upon treatment of cells with F-ara-A (100 μM, 72 hours), 15 nuclear proteins changed in abundance by more than 2-fold. Nuclear proteins up-regulated included calmodulin (4.3-fold), prohibitin (3.9-fold), β-actin variant (3.7-fold), and structure-specific recognition protein 1 (3.7-fold); those down-regulated included 60S ribosomal protein P2B (0.12-fold), fumarate hydratase (0.19-fold), splicing factor arginine/serine-rich 3 (0.35-fold), and replication protein A2 (0.42-fold). These changes in the levels of specific proteins promote survival or apoptosis; because the end result is apoptosis of MEC1 cells, apoptotic effects predominate.